Document ID: EPA-HQ-OPPT-2002-0043-0023
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2002-11-27T05:00Z

Unclassified
ENV/
JM/
RD(
2002)
17/
FINAL
Organisation
de
Coopération
et
de
Développement
Economiques
Organisation
for
Economic
Co­
operation
and
Development
21­
Nov­
2002
___________________________________________________________________________________________
English
­
Or.
English
ENVIRONMENT
DIRECTORATE
JOINT
MEETING
OF
THE
CHEMICALS
COMMITTEE
AND
THE
WORKING
PARTY
ON
CHEMICALS,
PESTICIDES
AND
BIOTECHNOLOGY
CO­
OPERATION
ON
EXISTING
CHEMICALS
HAZARD
ASSESSMENT
OF
PERFLUOROOCTANE
SULFONATE
(
PFOS)
AND
ITS
SALTS
JT00135607
Document
complet
disponible
sur
OLIS
dans
son
format
d'origine
Complete
document
available
on
OLIS
in
its
original
format
ENV/

JM/

RD(

2002)

17/

FINAL
Unclassified
English
­

Or.

English
ENV/
JM/
RD(
2002)
17/
FINAL
2
Preface
In
the
margins
of
the
ninth
meeting
of
the
Task
Force
on
Existing
Chemicals
(
29­
30
May
2000)
several
Member
countries
agreed
to
informally
work
together
to
collect
information
on
the
environmental
and
human
health
hazards
of
perfluorooctane
sulfonate
(
PFOS)
to
produce
a
hazard
assessment.
The
decision
followed
the
announcement
by
a
major
US
manufacturer
 
3M
 
to
globally
phase
out
the
manufacture
and
use
of
these
chemicals
beginning
in
2001.
The
US
and
the
UK
agreed
to
lead
the
activity
with
the
Secretariat
assisting
by
requesting
readily
available
exposure
information
from
Member
countries
as
well
as
from
non­
Member
countries
through
IFCS.

An
informal
meeting
was
hosted
by
the
US
on
26­
27
October
2000
(
Crystal
City,
Virginia,
US)
to:

 
review
the
current
status
of
assessment
activities;
 
learn
about
actions
being
taken
in
other
countries;
and
 
identify
planned
or
ongoing
work
on
this
issue.

In
preparation
for
the
meeting
3M
circulated
a
draft
initial
assessment
report
including
robust
study
summaries
of
key
studies,
together
with
exposure
information.

At
the
31st
Joint
Meeting
of
the
Chemicals
Committee
and
the
Working
Party
on
Chemicals,
Pesticides
and
Biotechnology
(
7­
10
November
2000),
it
was
agreed
that,
since
this
was
a
matter
of
sufficient
interest
to
all
Member
countries,
this
activity
should
be
undertaken
under
the
existing
Chemicals
Programme,
overseen
by
the
Task
Force.
As
PFOS
is
not
an
HPV
Chemical,
it
was
not
dealt
with
under
the
HPV
Chemicals
Programme.

A
draft
hazard
assessment
was
posted
on
the
OECD
web
site
for
comment
in
December
2000.
The
OECD
established
an
electronic
discussion
group
to
exchange
comments
and
information.
A
special
session
on
PFOS
and
its
salts
was
held
on
25
January
2001
in
Orlando,
USA,
as
part
of
the
11th
SIAM
meeting.
At
this
session
an
overview
of
the
draft
hazard
assessment
was
presented.

The
draft
hazard
assessment
was
revised
twice
since
December
2000
to
incorporate
comments
that
were
received,
as
well
as
to
incorporate
newly
completed
studies.
Comments
were
received
from
3M,
World
Wildlife
Fund,
Health
Canada,
Environment
Canada,
and
Australia.

At
the
11th
meeting
of
the
Task
Force
on
Existing
Chemicals
(
27­
28
May
2002),
the
revised
hazard
assessment
was
discussed.
The
Task
Force
agreed
with
the
conclusions
and
recommendations
of
the
hazard
assessment.
The
Task
Force
also
agreed
that
the
Secretariat
should
gather
information
from
governments
and
BIAC
on
risk
management
activities
currently
undertaken
or
planned
in
Member
countries
on
PFOS.

At
the
34th
Joint
Meeting
of
the
Chemicals
Committee
and
the
Working
Party
on
Chemicals,
Pesticides
and
Biotechnology
(
5­
8
November
2002),
the
final
draft
of
the
assessment
was
endorsed.
The
Joint
Meeting
recommended
that
this
document
be
derestricted
under
the
authority
of
the
Secretary
General.

This
hazard
assessment
of
perfluorooctane
sulfonate
(
PFOS)
and
its
salts
includes
all
information
that
was
available
by
July
2002.
A
quantitative
risk
assessment
was
not
conducted
as
this
should
entail
regional
exposure
information.
The
hazard
information
on
PFOS
should
be
used
with
caution
in
evaluating
the
potential
hazards
of
other
perfluorinated
compounds.
The
perfluorinated
compounds
represent
a
very
unique
chemistry
whose
toxicological
properties
are
presently
not
well
understood
and
clearly
the
presence
of
different
length
(
perfluorinated)
carbon
chains
and
functional
groups
are
likely
to
influence
toxicity.
It
is
not
clear
at
this
time
whether
the
hazard
concerns
of
PFOS
can
be
extrapolated
to
other
perfluorinated
compounds
except
under
circumstances
where
the
compound
may
degrade
to
PFOS.

Assessment
activities
on
PFOS
and
its
salts
are
also
on­
going
in
other
international
fora,
e.
g.
OSPAR.
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3
TABLE
OF
CONTENTS
Recommendations
of
the
Hazard
Assessment................................................................
5
Summary
and
Conclusions
of
the
Hazard
Assessment..................................................
5
1.0
Identity.............................................................................................................................
10
1.1
Physicochemical
Properties
...........................................................................................
10
2.0
General
Information
on
Exposure
................................................................................
11
2.1
Production
and
Use
of
PFOS.........................................................................................
12
2.1.1
PFOS­
Based
Surface
Treatment
Applications.............................................................
13
2.1.2
PFOS­
Based
Paper
Protection
Applications
................................................................
13
2.1.3
PFOS­
Based
Performance
Chemical
Applications......................................................
13
2.2
Environmental
Exposure
and
Fate................................................................................
15
2.2.1
Volatility
..........................................................................................................................
15
2.2.2
Combustion
.....................................................................................................................
15
2.2.3
Photolysis/
Oxidation.......................................................................................................
15
2.2.4
Biodegradation................................................................................................................
15
2.3
Environmental
Monitoring
............................................................................................
15
2.4
Human
Biomonitoring....................................................................................................
17
2.4.1
Occupational
Exposures.................................................................................................
17
2.4.2
Non­
occupational
Exposures
.........................................................................................
19
3.0
Human
Health
Hazards..................................................................................................
23
3.1
Metabolism
and
Pharmacokinetics
...............................................................................
23
3.1.1
Absorption.......................................................................................................................
23
3.1.2
Distribution
.....................................................................................................................
23
3.1.3
Elimination
......................................................................................................................
23
3.1.4
Half­
life
in
Humans
........................................................................................................
24
3.2
Acute
Toxicity
.................................................................................................................
25
3.3
Mutagenicity....................................................................................................................
26
3.4
Repeated
Dose
Toxicity
..................................................................................................
27
3.5
Carcinogenicity
...............................................................................................................
34
3.6
Developmental
Toxicity..................................................................................................
39
3.7
Reproductive
Toxicity
....................................................................................................
43
3.8
Human
Hazard................................................................................................................
50
4.0
Hazards
to
the
Environment..........................................................................................
55
4.1
Effects
on
Fish,
Invertebrates
and
Algae......................................................................
56
4.1.1
Fish
...................................................................................................................................
57
4.1.2
Invertebrates
...................................................................................................................
64
4.1.3
Aquatic
Plants
.................................................................................................................
69
4.2
Effects
on
Other
Aquatic
Organisms............................................................................
73
4.2.1
Amphibians
.....................................................................................................................
73
4.2.2
Sediment
Dwelling
Invertebrates
..................................................................................
75
4.2.3
Bacteria
............................................................................................................................
75
4.2.4
Activated
Sludge
Microorganisms.................................................................................
77
4.3
Effects
on
Terrestrial
Organisms
..................................................................................
79
4.3.1
Soil­
dwelling
Invertebrates
............................................................................................
79
4.3.2
Terrestrial
Plants
............................................................................................................
79
4.3.3
Birds
.................................................................................................................................
79
4.3.4
Bees...................................................................................................................................
81
5.0
References........................................................................................................................
83
ENV/
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4
Annex
1
Ecological
Studies
.................................................................................................
89
Annex
2
Robust
Summaries
of
Key
Ecotoxicology
Studies
.............................................
91
Annex
3
Application
of
Equilibrium
Partitioning
Models
to
Determining
Effect
Concentrations
for
PFOS
Salts
in
Soil
and
Sediment
.....................................
213
Annex
4
Summary
of
the
Lowest
Acceptable
Effect
Concentrations
...........................
216
Annex
5
Robust
Summaries
for
Physical
Chemical
Properties
and
Environmental
Fate
Studies
..............................................................................
218
Annex
6
Robust
Summaries
of
Toxicology
and
Human
Biomonitoring
Studies........................................................................................
256
ENV/
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5
RECOMMENDATIONS
OF
THE
HAZARD
ASSESSMENT
Perfluorooctane
sulfonate
(
PFOS)
is
a
candidate
for
further
work.

Sufficient
information
exists
to
address
hazard
classification
for
all
SIDS
human
health
endpoints.
PFOS
is
persistent,
bioaccumulative
and
toxic
to
mammalian
species.
There
are
species
differences
in
the
elimination
half­
life
of
PFOS;
the
half­
life
is
100
days
in
rats,
200
days
in
monkeys,
and
years
in
humans.
The
toxicity
profile
of
PFOS
is
similar
among
rats
and
monkeys.
Repeated
exposure
results
in
hepatotoxicity
and
mortality;
the
dose­
response
curve
is
very
steep
for
mortality.
This
occurs
in
animals
of
all
ages,
although
the
neonate
may
be
more
sensitive.
In
addition,
a
2­
year
bioassay
in
rats
has
shown
that
exposure
to
PFOS
results
in
hepatocellular
adenomas
and
thyroid
follicular
cell
adenomas;
the
hepatocellular
adenomas
do
not
appear
to
be
related
to
peroxisome
proliferation.
Further
work
to
elucidate
the
species
differences
in
toxicokinetics
and
in
the
mode
of
action
of
PFOS
will
increase
our
ability
to
predict
risk
to
humans.
Epidemiologic
studies
have
shown
an
association
of
PFOS
exposure
and
the
incidence
of
bladder
cancer;
further
work
is
needed
to
understand
this
association.

Sufficient
information
exists
to
address
hazard
classification
for
all
SIDS
environmental
endpoints.
PFOS
is
persistent
in
the
environment
and
has
been
shown
to
bioconcentrate
in
fish.
It
has
been
detected
in
a
number
of
species
of
wildlife,
including
marine
mammals.
Its
persistence,
presence
in
the
environment
and
bioaccumulation
potential
indicate
cause
for
concern.
It
appears
to
be
of
low
to
moderate
toxicity
to
aquatic
organisms
but
there
is
evidence
of
high
acute
toxicity
to
honey
bees.
No
information
is
available
on
effects
on
soil­
and
sediment­
dwelling
organisms
and
the
equilibrium
partitioning
method
may
not
be
suitable
for
predicting
PNECs
for
these
compartments.
PFOS
has
been
detected
in
sediment
downstream
of
a
production
site
and
in
effluents
and
sludge
from
sewage
treatment
plants.

Given
the
apparent
widespread
occurrence
of
PFOS,
national
or
regional
exposure
information
gathering
and
risk
assessment
may
need
to
be
considered.
In
addition,
data
on
its
toxicity
to
soil
and
sedimentdwelling
organisms
could
be
generated
as
a
post­
SIDS
activity.
There
is
currently
no
information
on
effects
on
soil­
or
sediment­
dwelling
organsisms
and
PFOS
has
been
detected
in
sediment
and
its
presence
in
sewage
sludge
could
lead
to
soil
exposure
if
spread
on
agricultural
land.

SUMMARY
AND
CONCLUSIONS
OF
THE
HAZARD
ASSESSMENT
Perfluorooctane
sulfonate
(
PFOS)
and
its
salts
are
fully
fluorinated
organic
compounds.
The
number
of
production
sites
is
not
clear,
but
there
is
production
in
the
US,
Europe
and
Japan.
In
recent
years
(
to
2000),
approximately
4,500
metric
tons
of
PFOS­
related
chemicals
have
been
produced
annually.
The
major
global
producer
of
PFOS
intends
to
cease
production
by
the
end
of
2002.

The
majority
of
PFOS­
related
chemicals
are
high
molecular
weight
polymers
in
which
PFOS
represents
a
fraction
of
the
total
molecular
weight.
PFOS­
related
chemicals
are
used
in
a
variety
of
products,
including
as
surface­
treatments
of
fabric
for
soil/
stain
resistance,
coating
of
paper
as
part
of
a
sizing
agent
formulation
and
in
specialised
applications
such
as
fire
fighting
foams.

PFOS
has
a
solubility
of
approximately
550
mg/
l
in
pure
water
at
24­
25
°
C.
The
solubility
decreases
significantly
with
increased
salt
content,
for
example
the
potassium
salt
of
PFOS
has
a
solubility
in
fresh
water
of
370
mg/
L
and
of
25
mg/
l
in
filtered
sea
water.
Due
to
the
surface­
active
properties
of
PFOS,
the
Log
Kow
cannot
be
measured.
The
potassium
salt
of
PFOS
has
a
low
vapour
pressure,
3.31
x
10­
4
Pa
at
20
°
C.
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6
Human
Health
In
human
blood
samples,
PFOS
has
been
detected
in
the
serum
of
occupational
and
general
populations.
In
the
U.
S.,
the
highest
reported
blood
serum
level
of
PFOS
was
observed
in
1995
in
a
manufacturing
employee
in
Decatur,
Alabama
(
12.83
ppm).
Mean
PFOS
levels
have
been
dropping
in
that
plant
and
a
plant
in
Belgium
since
1995,
the
most
recent
being
1.32
ppm
and
0.80
ppm,
respectively,
in
2000.
In
the
general
population,
serum
collected
from
blood
banks
and
commercial
sources
have
indicated
mean
PFOS
levels
of
30­
53
ppb.
In
individual
serum
samples
obtained
from
adults
and
children
in
various
regions
of
the
U.
S.,
mean
PFOS
levels
were
approximately
43
ppb.

Several
occupational
studies
have
been
conducted
on
volunteers
at
the
3M
plants
in
Decatur,
Alabama
and
Antwerp,
Belgium.
Cross­
sectional
studies,
based
on
the
results
of
a
voluntary
medical
surveillance
program
for
employees
at
each
plant,
did
not
report
consistent
associations
between
workers'
PFOS
levels
less
than
6
ppm
and
certain
hematology,
hormonal,
and
other
clinical
chemistry
parameters
in
1995
and
1997.
In
2000
when
the
analysis
included
male
employees
from
both
plants,
mean
values
for
triglycerides,
alkaline
phosphatase,
total
bilirubin,
and
ALT
were
significantly
(
p
<
.05)
higher
for
workers
with
the
highest
PFOS
serum
levels
(
1.69
 
10.06
ppm).
Serum
triiodothyronine
was
significantly
higher
and
thyroid
hormone
binding
ratio
was
significantly
lower
in
workers
with
the
highest
PFOS
serum
levels
(
p
<
.05
for
both).
The
association
with
T3
also
remained
significant
and
positive
in
multivariable
regression
analyses
adjusted
for
potential
confounders.
A
longitudinal
analysis
of
these
data
did
not
reveal
statistically
significant
associations
over
time
between
PFOS
and
cholesterol,
triglycerides,
and
other
lipid
and
hepatic
parameters.
Hormones
were
not
included
in
these
analyses.
There
are
several
limitations
to
both
the
cross­
sectional
and
longitudinal
studies,
such
as
the
voluntary
nature
of
the
medical
surveillance,
the
small
number
of
employees
participating
across
sampling
periods,
the
different
labs
and
analytical
techniques
used
to
measure
serum
PFOS,
and
the
differences
in
PFOS
levels,
demographics,
and
clinical
chemistries
between
employees
in
the
Decatur
and
Antwerp
plants.

In
a
mortality
study,
which
followed
workers
for
37
years,
mortality
risks
for
most
of
the
cancer
types
and
non­
malignant
causes
were
not
elevated.
However,
a
statistically
significant
risk
of
death
from
bladder
cancer
was
reported.
Three
male
employees
in
the
cohort
died
of
bladder
cancer
(
0.12
expected),
and
all
of
them
had
been
employed
at
the
plant
for
more
than
20
years.
All
of
them
had
also
worked
in
high
exposure
jobs
for
at
least
5
years.
In
order
to
screen
for
morbidity
outcomes,
an
"
episode
of
care"
analysis
was
undertaken
for
employees
who
had
worked
at
the
plant
between
1993
and
1998.
Many
different
types
of
cancer
and
other
non­
malignant
conditions
were
examined.
Increased
risks
were
not
reported
for
most
of
the
conditions
or
did
not
reach
statistical
significance.
However,
an
increased
risk
of
episodes
was
reported
for
neoplasms
of
the
male
reproductive
system,
the
overall
category
of
cancers
and
benign
growths,
and
neoplasms
of
the
gastrointestinal
tract.
These
risk
ratios
were
highest
in
employees
with
the
highest
and
longest
exposures
to
fluorochemicals.

Animal
studies
show
that
PFOS
is
well
absorbed
orally
and
distributes
mainly
in
the
serum
and
the
liver.
No
further
metabolism
is
expected.
Elimination
from
the
body
is
slow
and
occurs
via
the
urine
and
feces.
There
are
species
differences
in
the
elimination
half­
life
of
PFOS.
The
half­
life
in
serum
is
7.5
days
in
adult
rats
and
200
days
in
Cynomolgus
monkeys.
In
humans,
it
appears
to
be
quite
longer.
A
recent
halflife
analysis
was
conducted
on
9
retired
3M
chemical
workers.
PFOS
samples
were
collected
over
4
time
periods
spanning
180
days,
measured
in
triplicate
with
all
time
points
from
each
subject
analyzed
in
the
same
analytical
run.
The
mean
half­
life
for
PFOS
was
8.67
years
(
range
2.29
 
21.3
years,
SD
=
6.12).

PFOS
has
shown
moderate
acute
toxicity
by
the
oral
route
with
a
rat
LD50
of
251
mg/
kg.
A
one­
hour
LC50
of
5.2
mg/
l
in
rats
has
been
reported.
PFOS
was
found
to
be
mildly
irritating
to
the
eyes
and
nonirritating
to
the
skin
of
rabbits.
PFOS
has
not
been
shown
to
be
genotoxic
in
a
variety
of
assay
systems.
ENV/
JM/
RD(
2002)
17/
FINAL
7
Numerous
repeat­
dose
oral
toxicity
studies
on
PFOS
have
been
conducted
in
rats
and
primates.
In
general,
exposure
to
PFOS
results
in
hepatotoxicity
and
mortality;
the
dose­
response
curve
for
mortality
is
very
steep
for
rats
and
primates.
Adverse
signs
of
toxicity
observed
in
90­
day
rat
studies
included
increases
in
liver
enzymes,
hepatic
vacuolization
and
hepatocellular
hypertrophy,
gastrointestinal
effects,
hematological
abnormalities,
weight
loss,
convulsions,
and
death.
These
effects
were
reported
at
doses
of
2
mg/
kg/
day
and
above.
In
a
dietary
2­
year
bioassay
in
Sprague­
Dawley
rats,
hepatotoxicity,
characterized
by
centrilobular
hypertrophy,
centrilobular
eosinophilic
hepatocytic
granules,
centrilobular
hepatocytic
pigment,
or
centrilobular
hepatocytic
vacuolation
was
noted
in
male
and/
or
female
rats
given
5
or
20
ppm.
Hepatocellular
centrilobular
hypertrophy
was
also
observed
in
mid­
dose
(
2
ppm)
male
rats.
Significant
increases
in
the
incidence
of
cystic
hepatocellular
degeneration
were
found
in
all
the
male
treated
groups
(
0.5,
2,
5,
or
20
ppm).
Based
on
the
pathological
findings
in
the
liver,
the
LOAEL
was
5
ppm
and
the
NOAEL
was
2
ppm
in
female
rats.
In
males,
the
LOAEL
was
0.5
ppm,
and
a
NOAEL
was
not
established.

Adverse
signs
of
toxicity
observed
in
Rhesus
monkey
studies
included
anorexia,
emesis,
diarrhea,
hypoactivity,
prostration,
convulsions,
atrophy
of
the
salivary
glands
and
the
pancreas,
marked
decreases
in
serum
cholesterol,
and
lipid
depletion
in
the
adrenals.
The
dose
range
for
these
effects
was
reported
between
1.5­
300
mg/
kg/
day.
No
monkeys
survived
beyond
3
weeks
into
treatment
at
10
mg/
kg/
day
or
beyond
7
weeks
into
treatment
at
doses
as
low
as
4.5
mg/
kg/
day.
In
a
6­
month
study
of
Cynomolgus
monkeys,
low
food
consumption,
excessive
salivation,
labored
breathing,
hypoactivity,
ataxia,
hepatic
vacuolization
and
hepatocellular
hypertrophy,
significant
reductions
in
serum
cholesterol
levels,
and
death
were
observed
at
0.75
mg/
kg/
day.
No
effects
were
observed
at
doses
of
0.15
or
0.03
mg/
kg/
day.
No
effects
were
noted
in
animals
at
any
dose
level
following
a
52­
week
recovery
period.
The
average
concentration
of
PFOS
in
the
serum
following
26
weeks
of
treatment
was
11.1
+
1.52,
58.5
+
4.67
and
160
+
23.9
µ
g/
ml
for
the
females
in
the
0.03,
0.15
and
0.75
mg/
kg/
day
groups,
respectively;
for
males,
the
average
concentrations
were
15.9
+
5.54,
68.1
+
5.75
and
194
+
8.93
µ
g/
ml
in
the
0.03,
0.15
and
0.75
mg/
kg/
day
groups,
respectively.
After
the
52­
week
recovery
period,
the
serum
levels
were
21.4
+
2.01
and
41.4
+
1.15
µ
g/
ml
for
the
females
in
the
0.15
and
0.75
mg/
kg/
day
groups,
respectively;
for
males,
the
average
concentrations
were
19.1
+
0.805
and
41.1
+
25.9
µ
g/
ml
in
the
0.15
and
0.75
mg/
kg/
day
groups,
respectively.

The
potential
carcinogenicity
of
PFOS
has
been
examined
in
a
dietary
2­
year
bioassay
in
Sprague­
Dawley
rats.
There
was
a
significant
increase
in
the
incidence
of
hepatocellular
adenomas
in
males
and
females
at
the
highest
dose
of
20
ppm;
the
females
at
20
ppm
also
had
a
significant
increase
in
combined
hepatocellular
adenomas
and
carcinomas.
In
addition,
there
was
a
significant
increase
in
thyroid
follicular
cell
adenomas
and
combined
thyroid
follicular
cell
adenomas
and
carcinomas
in
the
male
recovery
group
at
20
ppm.
There
was
no
evidence
of
peroxisome
proliferation
in
the
livers
of
the
treated
animals.

Postnatal
deaths
and
other
developmental
effects
were
reported
at
low
doses
in
offspring
in
a
2­
generation
reproductive
toxicity
study
in
rats.
At
the
two
highest
doses
of
1.6
and
3.2
mg/
kg/
day,
pup
survival
in
the
first
generation
was
significantly
decreased.
All
first
generation
offspring
(
F1
pups)
at
the
highest
dose
died
within
a
day
after
birth
while
close
to
30%
of
the
F1
pups
in
the
1.6
mg/
kg/
day
dose
group
died
within
4
days
after
birth.
As
a
result
of
the
pup
mortality
in
the
two
top
dose
groups,
only
the
two
lowest
dose
groups,
0.1
and
0.4
mg/
kg/
day,
were
continued
into
the
second
generation.
The
NOAEL
and
LOAEL
for
the
second
generation
offspring
(
F2
pups)
were
0.1
mg/
kg/
day
and
0.4
mg/
kg/
day,
respectively,
based
on
reductions
in
pup
body
weight.

The
liver
and
serum
from
the
F0
and
F1
animals
was
analyzed
for
PFOS.
Qualitatively,
the
results
for
the
F0
animals
indicate
that
all
rats
(
including
controls)
had
detectable
levels
of
PFOS
in
serum
and
livers.
PFOS
concentration
increased
with
dose.
PFOS
concentrations
were
higher
in
the
liver
than
in
the
serum,
and
males
had
greatly
increased
PFOS
concentrations
in
serum
and
liver
when
compared
with
females
of
ENV/
JM/
RD(
2002)
17/
FINAL
8
the
same
dose
group.
Pooled
liver
samples
from
the
F1animals
sacrificed
shortly
after
birth
had
lower
PFOS
concentrations
than
adults
of
the
F0
generation
of
the
same
dose
group.

Based
on
the
results
of
the
two­
generation
reproductive
toxicity
study,
a
cross­
fostering
study
was
conducted
as
a
means
of
determining
whether
the
reductions
in
pup
viability
were
a
result
of
in
utero
exposure
to
PFOS
or
as
a
result
of
exposure
during
lactation;
thus
the
potential
for
a
distinction
to
be
made
between
prenatal
and
postnatal
effects
following
continuous
maternal
treatment.
Under
the
limited
conditions
of
the
study,
the
data
appear
to
indicate
that
reduced
pup
survival
is
mainly
a
result
of
in
utero
exposure
to
PFOS
and
that
post­
natal
exposure
via
milk
in
conjunction
with
in
utero
exposure
may
also
contribute
to
reduced
pup
survival.
In
contrast,
exposure
during
lactation
alone,
through
milk
from
exposed
dams,
does
not
appear
to
have
any
adverse
effect
on
pup
viability.

Several
mechanistic
studies
are
being
conducted
to
understand
the
neonatal
death
(
3M
Company,
2001c).
Preliminary
results
indicate
that
reductions
in
serum
lipids
and
cholesterol
synthesis
do
not
appear
to
play
a
significant
role
in
the
death
of
the
offspring.

Developmental
effects
were
also
reported
in
prenatal
developmental
toxicity
studies
in
the
rat
and
rabbit,
although
at
slightly
higher
dose
levels.
Signs
of
developmental
toxicity
in
the
offspring
were
evident
at
doses
of
5
mg/
kg/
day
and
above
in
rats
administered
PFOS
during
gestation.
Significant
decreases
in
fetal
body
weight
and
significant
increases
in
external
and
visceral
anomalies,
delayed
ossification,
and
skeletal
variations
were
observed.
A
NOAEL
of
1
mg/
kg/
day
and
a
LOAEL
of
5
mg/
kg/
day
for
developmental
toxicity
were
indicated.
In
the
same
study,
evidence
of
treatment­
related
signs
of
maternal
toxicity
were
also
observed
at
doses
of
5
mg/
kg/
day
and
above
and
mainly
consisted
of
hunched
posture,
anorexia,
bloody
vaginal
discharge,
uterine
stains,
alopecia,
rough
hair
coat,
and
bloody
crust,
as
well
as
decreases
in
body
weight
gains
and
food
consumption.
Reductions
in
the
mean
terminal
body
weights
minus
the
gravid
uterine
weights
were
also
observed
at
doses
>
5
mg/
kg/
day.
A
NOAEL
of
1
mg/
kg/
day
and
a
LOAEL
of
5
mg/
kg/
day
for
maternal
toxicity
were
indicated.
In
rabbits,
significant
reductions
in
fetal
body
weight
and
significant
increases
in
delayed
ossification
were
observed
in
the
offspring
of
pregnant
females
administered
PFOS
during
gestation
at
doses
of
2.5
mg/
kg/
day
and
above.
A
NOAEL
of
1.0
mg/
kg/
day
and
a
LOAEL
of
2.5
mg/
kg/
day
for
developmental
toxicity
were
indicated.
Maternal
toxicity
in
the
does
was
evident
at
doses
of
1.0
mg/
kg/
day
and
above,
and
consisted
of
an
increase
incidence
of
abortions
and
scant
feces,
as
well
as
significant
reductions
in
mean
maternal
body
weight
gains
and
food
consumption.
A
NOAEL
of
0.1
mg/
kg/
day
and
a
LOAEL
of
1.0
mg/
kg/
day
for
maternal
toxicity
were
indicated.

Environment
There
is
currently
little
information
on
the
life­
cycle
steps
that
may
lead
to
release
of
PFOS
to
the
environment.
However,
PFOS
has
been
detected
in
surface
water
and
sediment
downstream
of
a
production
facility
and
in
wastewater
treatment
plant
effluent,
sewage
sludge
and
landfill
leachate
at
a
number
of
cities
in
the
US.
Sampling
of
several
wildlife
species
from
a
variety
of
sites
across
the
United
States
has
shown
widespread
distribution
of
PFOS
and
it
was
detected
in
the
ppb
range
in
the
plasma
of
several
species
of
eagles,
wild
birds,
and
fish.
PFOS
has
been
detected
in
marine
mammals
at
a
number
of
locations
across
the
world.

PFOS
is
persistent
in
the
environment.
It
does
not
hydrolyse,
photolyse
or
biodegrade
under
environmental
conditions
and
is
not
expected
to
volatilize,
based
on
an
air/
water
partition
coefficient
of
<
2
E­
6
Pa.
m3/
mol.
PFOS
has
been
shown
to
bioconcentrate
in
the
tissues
of
bluegill
sunfish
and
carp.
In
bluegill
sunfish,
BCF
(
BCFK)
values
between
1124
and
4013
were
determined
and
PFOS
depurated
slowly
with
estimated
50%
clearance
times
of
up
to
116
days.
In
carp,
BCF
values
were
determined
to
be
between
200
and
1500.
ENV/
JM/
RD(
2002)
17/
FINAL
9
The
substance
shows
moderate
acute
toxicity
to
aquatic
organisms,
the
lowest
LC50
for
fish
is
a
96­
hour
LC50
of
4.7
mg/
l
to
the
fathead
minnow
Pimephales
promelas
for
the
lithium
salt.
For
aquatic
invertebrates,
the
lowest
EC50
for
freshwater
species
is
a
48­
hour
EC50
of
27
mg/
l
for
Daphnia
magna
and
for
saltwater
species,
a
96­
hour
LC50
value
of
3.6
mg/
l
for
the
Mysid
shrimp
Mysidopsis

bahia.
Both
tests
were
conducted
on
the
potassium
salt.
For
algae,
the
potassium
salt
gave
a
96h
NOEC
of
>
3.2
mg/
l
with
Skeletonema
costatum.

Long­
term
toxicity
data
is
available
for
fish
and
aquatic
invertebrates.
The
lowest
NOEC
for
fish
is
a
42
day
NOEC
(
survival)
of
0.3
mg/
l
in
an
early
life
stage
test
with
Pimephales
promelas
using
the
potassium
salt.
The
lowest
NOEC
for
aquatic
invertebrates
is
a
35­
day
NOECreproduction
of
0.25
mg/
l
for
Mysidopsis
bahia
using
the
potassium
salt.
For
freshwater
species,
there
is
a
28­
day
NOECreproduction
of
7
mg/
l
for
Daphnia
magna,
also
using
the
potassium
salt.

A
growth
inhibition
test
has
been
carried
out
on
PFOS
potassium
salt
with
Lemna
gibba
(
Duckweed).
The
test
gave
a
7­
day
IC50
of
108
mg/
l
for
inhibition
of
frond
production
and
a
7­
day
NOEC
of
15.1
mg/
l
based
on
the
inhibition
of
frond
production
and
evidence
of
sub­
lethal
effects.

PFOS
does
not
appear
to
be
toxic
to
sewage
sludge
microorganisms.
In
an
activated
sludge
respiration
inhibition
test,
the
3­
hour
IC50
value
for
PFOS
(
potassium
salt)
was
>
905
mg/
l
(
nominal
concentration).

No
data
are
available
for
effects
on
soil­
dwelling
or
sediment­
dwelling
species.
The
use
of
equilibrium
partitioning
models
to
derive
a
PNEC
for
these
compartments
may
not
be
applicable
to
this
anionic
surfactant.

PFOS
has
been
tested
on
two
species
of
bird,
the
Mallard
duck,
Anas
platyrhynchos,
and
the
Northern
Bobwhite
quail,
Colinus
virginianus.
The
lowest
acute
dietary
LC50
value
of
220
mg/
kg
of
food
was
determined
in
the
test
with
the
quail.
The
lowest
NOEC
of
37
mg/
kg
of
food
for
effects
on
body
weight
was,
in
contrast,
obtained
in
the
test
with
the
duck.

There
are
data
available
from
acute
oral
and
contact
toxicity
tests
on
the
Honey
bee
(
Apis
mellifera)
using
PFOS
potassium
salt.
These
studies
indicate
moderate
and
high
orders
of
toxicity
of
PFOS
to
bees
when
administered
via
these
routes.
The
acute
oral
test
yielded
a
72­
hour
LD50
for
ingestion
of
PFOS
of
0.40
µ
g/
bee
and
a
72­
hour
NOEL
of
0.21
µ
g/
bee.
The
contact
test
yielded
a
96­
hour
LD50
of
4.78
µ
g/
bee
and
a
96­
hour
NOEL
of
1.93
µ
g/
bee.

The
results
of
an
amphibian
teratogenesis
study
carried
out
with
Xenopus
laevis
(
African
clawed
frog)
show
PFOS
potassium
salt
to
be
acutely
toxic
to
(
96­
hour
LC50
=
13.8
mg/
l),
and
cause
malformations
in
(
96­
hour
EC50
=
12.1
mg/
l),
frog
embryos.
The
minimum
concentration
that
inhibited
growth
of
the
embryos
was
determined
to
be
7.97
mg/
l.
A
teratogenic
index
of
1.1
was
determined
from
the
ratio
of
the
96­
hour
LC50
to
the
96­
hour
EC50,
indicating
a
low
potential
for
PFOS
to
be
a
developmental
hazard
in
this
species.
ENV/
JM/
RD(
2002)
17/
FINAL
10
1.0
Identity
Chemical
Name:
Perfluorooctane
Sulfonate
The
perfluorooctane
sulfonate
anion
(
PFOS)
does
not
have
a
specific
CAS
number.
The
acid
and
salts
have
the
following
CAS
numbers:

acid
(
1763­
23­
1)
ammonium
(
NH4
+)
salt
(
29081­
56­
9)
diethanolamine
(
DEA)
salt
(
70225­
14­
8)
potassium
(
K+)
salt
(
2795­
39­
3)
lithium
(
Li+)
salt
(
29457­
72­
5)

Molecular
formula:
C8F17SO3
Structural
formula:
CF3­
CF2­
CF2­
CF2­
CF2­
CF2­
CF2­
CF2­
S(=
O)(=
O)

OSynonyms
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro;
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­
1­
octanesulfonic
acid;
1­
Octanesulfonic
acid,
heptadecafluoro­;
1­
Perfluorooctanesulfonic
acid;
Hepatadecafluoro­
1­
octanesulfonic
acid;
Perfluoro­
n­
octanesulfonic
acid;
Perfluorooctanesulfonic
acid;
Perfluorooctylsulfonic
acid
1.1
Physicochemical
Properties
Due
to
the
surface­
active
properties
of
PFOS
and
the
test
protocol
itself,
PFOS
forms
three
layers
in
octanol/
water
and
hence,
an
n­
octanol/
water
(
Kow)
partition
coefficient
cannot
be
determined.
Consequently,
the
various
physicochemical
properties
(
e.
g.,
bioconcentration
factor,
soil
adsorption
coefficient),
which
can
usually
be
estimated
for
conventional
organic
compounds
utilizing
Kow
equations,
cannot
be
estimated,
and
a
calculated
(
estimated)
log
Kow
cannot
be
trusted.
Even
if
the
log
Kow
were
known,
it
may
not
be
appropriate
for
predictive
purposes,
e.
g.,
bioconcentration.
Studies
on
laboratory
rats
indicate
that
PFOS
does
not
bioconcentrate
in
the
lipid
fraction.
Instead,
it
tends
to
bind
to
certain
proteins.
In
two
studies,
PFOS
was
reported
to
have
a
mean
solubility
of
519
mg/
L
and
570
mg/
L
in
pure
water
at
24­
25
°
C.
Solubility
decreases
significantly
with
increased
salt
content
(
12.4
mg/
L
in
natural
seawater
at
22­
23
°
C,
and
20.0
mg/
L
in
a
3.5%
NaCl
solution
at
22­
24
°
C
(
3M
Company,
2001a).
In
a
related
study,
PFOS
was
reported
to
have
a
mean
solubility
of
56.0
mg/
L
in
pure
octanol
(
3M
Company,
2001b).
These
data
suggest
that
any
PFOS
discharged
to
a
water
source
would
tend
to
remain
in
that
medium,
unless
it
is
adsorbed
onto
particulate
matter
or
assimilated
by
organisms.
If
PFOS
does
bind
to
particulate
matter
the
material
would
ultimately
end
up
in
the
sediment.
Further
study
is
underway
to
determine
the
presence
of
PFOS
in
sediments
from
various
locations
and
the
binding
potential
of
PFOS
to
sediments.

The
available
physicochemical
properties
for
the
potassium
salt
of
PFOS
are
as
follows
(
3M
Report,
1999):

Melting
point:
>=
400
°
C
Boiling
point:
not
calculable
Vapor
pressure:
3.31
x
10­
4
Pa
at
20
°
C
(
3.27
x
10­
9
atm)
ENV/
JM/
RD(
2002)
17/
FINAL
11
Air/
water
partition
coefficient
in
pure
water:
0
(<
2
x
10­
6
)

Solubility:
pure
water:
570
mg/
L
Solubility:
fresh
water:
370
mg/
L
Solubility:
unfiltered
seawater:
12.4
mg/
L
Solubility:
filtered
seawater:
25
mg/
L
The
specific
gravities
(
water
=
1)
and
pH
values
(
in
parentheses)
of
the
PFOS
salts
are
as
follows
(
3M
Report,
1999):

DEA,
~
1.1
(~
7)
NH4+
,
~
1.1
(~
7)
Li+,
~
1.1
(
6­
8)
K+,
~
0.6
(
7­
8)

The
above
water
solubility
values
correspond
to
the
following
Henry's
Law
constants
(
atm.
m3/
mol),
calculated
herein,
utilizing
the
vapor
pressure
of
3.27
x
10­
9
atm
given
in
the
3M
Report
(
1999):

Pure
water:
3.05
x
10­
9
Fresh
water:
4.7
x
10­
9
Unfiltered
seawater:
1.4
x
10­
7
Filtered
seawater:
2.4
x
10­
8
As
a
point
of
reference,
the
Henry's
Law
constant
for
pure
water
at
20
°
C
is
4.34
x
10­
7
.

2.0
General
Information
on
Exposure
The
starting
material
for
PFOS­
related
chemicals
is
perfluorooctanesulfonyl
fluoride
(
POSF).
POSF
is
manufactured
through
a
process
known
as
Simons
Electro­
Chemical
Fluorination
(
ECF)
in
which
an
electric
current
is
passed
through
a
solution
of
anhydrous
hydrogen
fluoride
and
an
organic
feedstock
of
1­
octanesulfonyl
fluoride.
The
ECF
process
replaces
the
carbon­
hydrogen
bonds
on
molecules
of
the
organic
feedstock
with
carbon­
fluorine
bonds.
Perfluorination
occurs
when
all
the
carbon­
hydrogen
bonds
are
replaced
with
carbon­
fluorine
bonds.
The
ECF
process
yields
between
30­
45
percent
straight
chain
(
normal)
POSF,
along
with
a
variable
mixture
of
byproducts
and
impurities.
The
output
of
the
ECF
process
is
not
a
pure
chemical,
but
instead
a
mixture
of
isomers
and
homologues
including
higher
and
lower
straight­
chain
homologues;
branched­
chain
perfluoroalkyl
fluorides
of
various
chain
lengths;
straightchain
branched,
and
cyclic
perfluroalkanes
and
ethers;
and
other
byproducts
(
3M
Company,
2000b).

According
to
information
available
to
the
U.
S.
Environmental
Protection
Agency
(
USEPA),
3M
Company
is
the
dominant
producer
of
POSF.
In
1997,
3M
reported
the
manufacture
or
importation
into
the
United
States
of
approximately
1,848
metric
tons
of
POSF.
For
2000,
3M
forecasts
a
volume
of
1,820
metric
tons
manufactured
or
imported
into
the
United
States.
After
accounting
for
3M
operations
in
Antwerp,
Belgium,
3M
estimates
a
total
global
POSF
production
of
3,665
metric
tons
for
2000
(
3M
Company,
2000d).
Production
of
POSF
by
3M
is
expected
to
decline
to
zero
by
the
end
of
2002
as
3M
scales
back
the
production
POSF­
derived
chemicals
(
3M
Company,
2000d).

Since
most
POSF
is
incorporated
into
higher
molecular
weight
polymers,
it
comprises
only
a
portion
of
the
mole
fraction
of
the
entire
polymer
in
the
final
product.
However,
it
should
be
noted
that
the
secondary
reactions
used
to
produce
POSF
derivatives
do
not
necessarily
produce
pure
products.
Typically,
1­
2%
of
the
final
product
is
comprised
of
unreacted
or
partially
reacted
fluorinated
starting
materials
or
intermediates
that
are
carried
forward
into
the
final
product
as
impurities
(
3M
Company,
2000b).
ENV/
JM/
RD(
2002)
17/
FINAL
12
To
date,
3M
has
not
provided
information
on
the
total
cumulative
production
volumes
of
POSF
or
PFOSrelated
chemicals
since
initial
commercialization
over
40
years
ago.
Precise
production
volume
information
for
manufacturers
other
than
3M
outside
the
United
States
has
also
been
difficult
to
obtain.
3M,
however,
has
asserted
that
it
is
the
dominant
global
producer
of
PFOS
chemicals,
responsible
for
the
large
majority
of
total
global
production
volumes
(
3M
Company,
2000e).
The
limited
production
volume
information
provided
by
OECD
member
countries
for
companies
located
outside
the
United
States
supports
this
conclusion.

Aside
from
the
United
States
and
Belgium,
other
OECD
Member
countries
that
reportedly
have
production
capacity
include
Italy
and
Japan.
There
may
also
be
some
production
in
non­
OECD
countries.
Following
are
companies
that
have
been
identified
in
various
chemical
buyer's
guides
as
offering
PFOS­
related
chemicals
for
sale
(
Directory
of
World
Chemical
Producers,
2000;
ChemSources
USA,
2000;
OPD
Chemical
Buyers
Directory,
2000).
This
information
has
not
been
corroborated
independently,
except
for
Miteni
S.
p.
A.
of
Italy
and
Dianippon
Ink
&
Chemicals,
Inc.
of
Japan.

OECD
member
countries
°
Miteni
S.
p.
A.
(
Italy)
°
EniChem
Synthesis
S.
p.
A.
(
Italy)
°
Dianippon
Ink
&
Chemicals,
Inc.
(
Japan)
°
Midori
Kaguka
Co.,
Ltd.
(
Japan)
°
Tohkem
Products
Corporation
(
Japan)
°
Tokyo
Kasei
Kogyo
Company,
Ltd.
(
Japan)
°
Fluka
Chemical
Co,
Ltd.
(
Switzerland)
°
BNFL
Fluorochemicals
Ltd.
(
United
Kingdom)
°
Fluorochem
Ltd.
(
United
Kingdom)

Non­
OECD
countries
°
Milenia
Agro
Ciencias
S.
A.
(
Brazil)
°
Changjiang
Chemical
Plant
(
China)
°
Indofine
Chemical
Company,
Inc.
(
India)
°
Scientific
Industrial
Association
P
&
M
Ltd.
(
Russian
Federation)

2.1
Production
and
Use
of
PFOS
The
majority
of
PFOS­
related
chemicals
are
high
molecular
weight
polymers
in
which
PFOS
represents
a
fraction
of
the
total
molecular
weight.
PFOS­
related
chemicals
are
used
in
a
variety
of
products,
as
shown
in
the
figure
following
this
section.
These
products
can
be
divided
into
three
main
categories
of
use:

°
Surface
treatments,
°
Paper
protection,
and
°
Performance
chemicals.

Each
of
these
categories
and
the
associated
3M
production
volume
is
described
in
more
detail
below.
Note
that
in
many
cases
the
production
volumes
represent
total
solid
metric
tons
of
fluorochemical­
containing
compound,
not
PFOS
itself.
Less
than
91
metric
tons
of
PFOS
and
its
salts
are
commercialized
as
finished
products
(
3M,
2000b).

In
addition,
varying
amounts
of
fluorochemical
residuals
(
unreacted
or
partially
reacted
starting
materials
ENV/
JM/
RD(
2002)
17/
FINAL
13
or
intermediates)
are
carried
forward
into
final
products
at
concentrations
of
1­
2%
or
less
as
a
result
of
the
ECF
manufacturing
process.
These
residuals
also
have
the
potential
to
degrade
or
metabolize
to
PFOS
(
3M,
2000b).

2.1.1
PFOS­
Based
Surface
Treatment
Applications
PFOS­
related
chemicals
produced
for
surface
treatment
applications
provide
soil,
oil,
and
water
resistance
to
personal
apparel
and
home
furnishings.
Specific
applications
in
this
use
category
include
protection
of
apparel
and
leather,
fabric/
upholstery,
and
carpet.
These
applications
are
undertaken
in
industrial
settings
by
customers
such
as
textile
mills,
leather
tanneries,
finishers,
fiber
producers,
and
carpet
manufacturers.
PFOS­
related
chemicals
are
also
used
in
aftermarket
treatment
of
apparel
and
leather,
upholstery,
carpet,
and
automobile
interiors
by
the
general
public
or
professional
applicators
(
3M
Company,
2000b).
In
2000,
the
global
production
volume
of
PFOS­
related
chemicals
for
this
use
category
is
estimated
at
approximately
2,160
metric
tons
(
3M
Company,
2000b).

2.1.2
PFOS­
Based
Paper
Protection
Applications
PFOS­
related
chemicals
produced
for
paper
protection
applications
provide
grease,
oil,
and
water
resistance
to
paper
and
paperboard
as
part
of
a
sizing
agent
formulation.
Specific
applications
in
this
use
category
include
food
contact
applications
(
plates,
food
containers,
bags,
and
wraps),
as
well
as
non­
food
contact
applications
(
folding
cartons,
containers,
carbonless
forms,
and
masking
papers).
The
application
of
sizing
agents
is
undertaken
mainly
by
paper
mills
and,
to
some
extent,
converters
who
manufacture
bags,
wraps,
and
other
products
from
paper
and
paperboard
(
3M
Company,
2000d).
In
2000,
the
global
production
volume
of
PFOS­
related
chemicals
for
this
use
category
is
estimated
at
approximately
1,490
metric
tons
(
3M
Company,
2000b).

2.1.3
PFOS­
Based
Performance
Chemical
Applications
PFOS­
related
chemicals
in
the
performance
chemical
category
are
used
in
a
variety
of
specialized
industrial,
commercial,
and
consumer
applications.
This
category
includes
various
salts
of
PFOS
that
are
commercialized
as
finished
products.
Specific
applications
in
this
category
include
fire
fighting
foams,
mining
and
oil
well
surfactants,
acid
mist
suppressants
for
metal
plating
and
electronic
etching
baths,
photolithography,
electronic
chemicals,
hydraulic
fluid
additives,
alkaline
cleaners,
floor
polishes,
photographic
film,
denture
cleaners,
shampoos,
chemical
intermediates,
coating
additives,
carpet
spot
cleaners,
and
as
an
insecticide
in
bait
stations
(
3M
Company,
2000b;
Boeing,
2000).
In
2000,
the
global
production
volume
of
PFOS
chemicals
for
this
use
category
is
estimated
at
approximately
831
metric
tons
(
3M
Company,
2000d).
Of
this
volume,
approximately
151
metric
tons
will
be
used
in
fire
fighting
foams.
ENV/
JM/
RD(
2002)
17/
FINAL
14
Perfluorooctanesulfonyl
fluoride
(
PFOSF)
chemical
intermediate
Perfluorooctanesulfonic
acid
(
PFOSA)
chemical
intermediate
acid
catalyst
for
photoresists
K,
Li,
DEA,
NH
4
Salts
surfactant
in
fire
fighting
foam
surfactant
for
alkaline
cleaners
emulsifier
in
floor
polish
mist
suppressant
for
metal
plating
baths
surfactant
for
etching
acids
for
circuit
boards
pesticide
active
ingredient
for
ant
bait
traps
N­
Alkylperfluorooctanesulfonamide
(
FOSA)
chemical
intermediate
pesticide
active
ingredient
Alkoxylates
Carboxylates
antistatic
agent
in
photographic
paper
Oxazolidinones
waterproofing
casts/
wound
dressings
Amides
pesticide
active
ingredient
Quaternary
Ammonium
Salts
mist
suppressant
for
metal
plating
baths
Amines
mist
suppressant
for
metal
plating
baths
Amphoterics
water/
solvent
repellency
for
leather/
paper
Silanes
N­
Alkylperfluorooctanesulfonamidoethanol
(
FOSE)
chemical
intermediate
Acrylates
Adipates
Phosphate
Esters
Fatty
Acid
Esters
Alcohols
Urethanes
Copolymers
Electro­
Chemical
Fluorination
(
ECF)
Cells
(
Octanesulfonyl
fluoride
+
HF
+
electricity)

soil/
oil/
water
repellency
for:
carpet
fabric/
upholstery
apparel
leather
metal/
glass
oil/
water
repellency
for:
plates
food
containers
bags
wraps
folding
cartons
containers
carbonless
forms
masking
papers
Polyesters
Perfluorooctyl
Sulfonates:
Major
Product
Categories
ENV/
JM/
RD(
2002)
17/
FINAL
15
2.2
Environmental
Exposure
and
Fate
PFOS
does
not
hydrolyze,
photolyze
or
biodegrade
under
environmental
conditions
and
it
is
persistent
in
the
environment.

2.2.1.
Volatility
PFOS
is
not
expected
to
volatilize,
based
on
an
assigned
air/
water
partition
coefficient
of
<
2
x
10­
6
Pa.
m3/
mol.
This
assignment
was
made
by
Prof.
Donald
Mackay
based
on
experimental
data
generated
by
3M.
In
that
study,
the
Hc
was
too
small
to
measure
experimentally
using
pure
water.
The
interpretation
by
Mackay
states
"[
PFOS]
is
thus
essentially
non­
volatile
from
aqueous
solution.
This
is
probably
because
of
its
ionic
nature.
The
simple
expedient
is
to
assign
it
a
Kaw
of
zero,
i.
e.
is
a
type
2
involatile
chemical
in
our
nomenclature."
This
determination
is
in
agreement
with
the
Henry's
Law
constant
of
3.05
x
10­
9
calculated
herein
for
pure
water.
Overall,
the
conclusion
is
that
PFOS
is
a
substance
with
very
low
and
possibly
negligible
volatility.

2.2.2
Combustion
To
begin
to
understand
the
potential
degradation
pathways
for
perfluorinated
chemicals,
the
incineration
process
was
thermodynamically
modeled.
The
most
pertinent
finding
of
the
reported
analysis
is
that
the
carbon­
sulfur
bond
in
the
perfluorooctane
sulfonate
molecule
is
a
fairly
weak
bond.
To
validate
the
models
prediction,
laboratory
combustion
studies
on
a
series
of
perfluoroalkyl
compounds,
including
PFOS
and
two
polymeric
product
formulations
are
being
conducted.
An
assessment
will
be
made
of
combustion
by­
products
through
a
range
of
temperatures.
These
data
will
be
important
to
determine
whether
PFOS
may
enter
the
atmosphere
as
a
result
of
incomplete
combustion
of
waste.

2.2.3
Photolysis/
Oxidation
PFOS
does
not
appear
to
photolyze
(
Hatfield,
T.
2001).
Screening
studies
on
the
aqueous
photolytic
degradation
of
PFOS,
EtFOSE
alcohol,
MeFOSE
alcohol,
EtFOSA
and
MeFOSA
as
well
as
a
surfactant
and
foamer
product,
all
appeared
to
undergo
indirect
photolysis
to
FOSA,
PFOA,
a
hydride
and
olefins;
PFOS
was
not
detected
(
Hatfield,
T.
2001,
3M
Report
No.
W2775).
One
product,
an
aromatic
perfluorooctane
sulfonate,
did
photodegrade
to
form
PFOS
(
3M
Report,
1999).

2.2.4
Biodegradation
The
biodegradability
of
PFOS
was
examined
in
a
MITI­
I
test
(
Kurume
Laboratory
(
2002).
No
significant
degradation
of
PFOS
was
observed
in
28
days,
either
as
net
oxygen
demand
from
degradation
of
test
substance
(
i.
e.,
ultimate
degradation),
loss
ot
total
organic
carbon
(
TOC;
another
way
to
measure
ultimate
degradation),
or
loss
of
parent
compound
identity
(
primary
degradation).
Average
percentage
biodegradation
after
28
days
was
observed
to
be
0%
by
oxygen
demand;
6%
by
removal
of
total
organic
carbon;
and
3%
by
liquid
chromatography­
mass
spectrometry
(
LC­
MS;
measures
primary
degradation).
These
values
are
indicative
of
no
significant
degradation,
within
the
accuracy
limits
for
this
test.
Validity
of
the
test
was
confirmed
by
degradation
of
the
control
substance
aniline,
for
which
%
of
theoretical
oxygen
demand
exceeded
the
pass
criterion
of
45%
after
7
days
and
60%
after
14
days
respectively
(
74%
and
85%
of
theoretical
at
days
7
and
14,
respectively).

2.3
Environmental
Monitoring
3M's
Multi­
City
Study
reported
on
PFOS
concentrations
from
water,
sludge,
sediment,
POTW
effluent
and
landfill
leachate
samples
taken
in
six
cities
(
3M,
2001a).
Four
of
the
cities
(
Decatur
AL,
Mobile
AL,
ENV/
JM/
RD(
2002)
17/
FINAL
16
Columbus
GA,
Pensacola
FL)
were
supply
cities
that
have
manufacturing
or
industrial
use
of
fluorochemicals;
two
of
the
cities
(
Cleveland
TN,
Port
St.
Lucie
FL)
were
control
cities
that
do
not
have
significant
fluorochemical
activities.
Across
all
cities,
POTW
effluent
concentrations
ranged
from
0.041
to
5.29
ppb.
The
POTW
sludge
(
dry
wt.)
range
was
less
than
0.2
ug/
kg
to
3,120
ppb;
the
drinking
water
range
was
non­
detect
to
0.063
ppb;
the
landfill
leachate
range
was
non­
detect
to
53.1
ppb;
the
surface
water
range
was
non­
detect
to
0.138
ppb;
the
sediment
range
was
non­
detect
to
1.13
ppb
(
dry
wt.);
and
the
quiet
water
range
was
non­
detect
to
2.93
ppb.
The
control
cities
samples
generally
inhabited
the
lower
end
of
the
above
ranges,
except
for
the
POTW
effluent
and
sludge
findings
for
Cleveland,
which
were
intermediate
in
their
ranges,
and
the
quiet
water
samples
at
Port
St.
Lucie,
which
were
the
highest.
3M
states
that
all
the
PFOS
surface
water
concentrations
are
below
laboratory­
derived
NOEC
(
No
Observed
Effect
Concentrations)
for
aquatic
organisms.

The
Multi­
City
Study
also
included
a
market
basket
sampling
of
PFOS
residue
in
a
total
of
over
200
samples
taken
from
green
beans,
apples,
pork
muscle,
cow
milk,
chicken
muscle,
chicken
eggs,
bread,
hot
dogs,
catfish,
and
ground
beef
(
3M,
2001a).
Measurable
quantities
of
PFOS,
to
0.852
ng/
g,
were
found
in
four
milk
samples
and
one
ground
beef
sample;
one
of
the
four
milk
samples
was
from
a
control
city,
the
balance
of
the
samples
with
measurable
PFOS
were
from
cities
with
fluorochemical
manufacture
or
use.

Giesy
reported
the
results
of
a
global
monitoring
survey
of
PFOS
in
marine
mammals;
they
were
located
in
Florida,
California
and
Alaskan
U.
S.
coastal
waters,
the
northern
Baltic
Sea,
the
Mediterranean
Sea,
the
Arctic
(
Spitsbergen),
and
Sable
Island
in
Canada.
PFOS
was
detected
in
the
liver
and
blood
of
marine
mammals
from
most
locations.
The
largest
liver
concentration
was
1520
ng/
g,
wet
wt.,
in
bottlenose
dolphin
from
Florida;
the
largest
blood
concentration
was
475
ng/
mL
in
ringed
seal
from
the
northern
Baltic
Sea.

The
plasma
of
piscivorous
(
fish­
eating)
birds
was
sampled
in
the
late
1980'
s
and
early
1990'
s
or
obtained
from
the
U.
S.
Fish
and
Wildlife
Service
(
date
of
liver
samples
was
not
listed),
and
these
samples
were
later
sent
to
3M
for
analysis
of
PFOS
levels
(
3M
Company,
2000a).
Species
sampled
for
PFOS
plasma
levels
included
the
bald
eagle,
albatross,
and
sea
eagle.
Plasma
levels
ranged
from
below
the
limit
of
detection
(
1
ppb)
up
to
a
maximum
of
1047
ppb,
present
in
a
bald
eagle.
Levels
in
the
livers
sampled
from
six
bird
species
ranged
from
below
the
limit
of
detection
up
to
a
maximum
of
2055
ppb.

Giesy
reported
on
PFOS
in
fish
and
fish­
eating
water
birds.
Fish
were
sampled
from
the
U.
S.,
certain
European
countries,
the
North
Pacific
Ocean
and
Antarctic
locations
(
Giesy,
2001a).
The
highest
concentration
was
923
ng/
g,
wet
wt.,
in
the
muscle
of
fish
from
a
Belgium
estuary.
The
highest
concentration
of
PFOS
in
U.
S.
Great
Lakes
fish
was
297
ng/
g,
wet
wt.,
in
the
muscle
of
carp.
Muscle
tissue
of
fish
from
several
inland
lakes
in
Michigan
U.
S.
did
not
contain
detectable
PFOS,
nor
did
fish
from
the
North
Pacific
and
Antarctic
oceans.
Fish­
eating
bird
samples
were
collected
from
the
U.
S.,
including
Midway
atoll,
the
Baltic
and
Mediterranean
Seas,
Japanese
and
Korean
coasts
(
Giesy,
2001b).
PFOS
concentrations
collected
from
the
plasma
of
bald
eagles
in
the
midwestern
USA
ranged
from
not
quantifiable
to
2220
ng/
mL,
with
a
mean
of
330
ng/
mL.
The
largest
PFOS
concentration
in
liver
samples
from
USA
birds
was
1780
ng/
g,
wet
wt.
from
a
Brandts
cormorant.
PFOS
concentrations
in
the
sera
of
North
Pacific
Ocean
albatrosses
ranged
from
3
to
34
ng/
mL.
PFOS
concentrations
in
blood
or
livers
of
birds
from
the
Mediterranean
and
Baltic
Seas
were
relatively
less
than
those
found
in
U.
S.
birds.

Giesy
reported
on
PFOS
in
mink
and
river
otter
livers
from
the
U.
S.
(
Giesy,
2001c).
PFOS
was
found
in
all
samples,
with
the
largest
concentration
of
4800
ng/
g,
wet
wt.,
found
in
mink
liver.
PFOS
concentrations
in
river
otter
livers
from
Washington
and
Oregon
states
ranged
from
34
to
994
ng/
g,
wet
wt.

Giesy
reported
that
PFOS
was
detected
in
oysters
collected
in
the
Chesapeake
Bay
and
Gulf
of
Mexico
of
the
U.
S.
coast
at
51
of
77
sampling
locations
at
a
concentration
range
of
<
42
to
1225
ng/
g,
dry
wt.
(
Giesy,
ENV/
JM/
RD(
2002)
17/
FINAL
17
2001d).

Giesy
reported
on
the
concentrations
of
PFOS
in
surface
water,
sediments,
clams,
and
fish
collected
from
locations
upstream
and
downstream
of
the
3M
facility
at
Decatur
AL
(
Giesy,
2001e).
Of
the
three
downstream
sampling
locations,
the
two
closest
to
the
3M
facility
had
PFOS
surface
water
concentrations
significantly
greater
than
the
two
upstream
sites
(
means
of
150
ug/
L
and
82
ug/
L,
vs.
0.009
(
est.)
and
0.053
ug/
L);
the
nearest
two
locations
had
sediment
concentrations
significantly
greater
than
the
upstream
sites
(
wet
wt.
means
5930
ug/
kg,
1299
ug/
kg
vs.
0.18(
est.)
and
0.98).
Clam
and
fish
samples
were
collected
at
two
locations,
one
upstream
and
one
downstream
of
the
3M
facility.
The
average
fish
whole
body
PFOS
concentration
for
the
upstream
location
was
59.1
ug/
kg
(
wet
wt.),
while
that
for
the
downstream
location
was
1332
ug/
kg.
The
average
PFOS
concentration
in
clams
at
the
upstream
location
was
15.6(
est.)
ug/
kg;
that
for
the
downstream
location
was
14.1(
est.)
ug/
kg.

Hansen
(
2002)
reported
concentrations
of
PFOS
measured
from
surface
water
samples
taken
from
the
Tennessee
River
up­
and
downstream
of
the
outfall
from
the
fluorochemical
manufacturing
facility
at
Decatur
AL
(
the
3M
facility
mentioned
above).
There
were
20
sampling
sites
above
and
20
sites
below
the
outfall
location,
spaced
at
approximately
2
mile
intervals.
Upstream
of
the
facility
the
average
concentration
of
PFOS
was
32
+
11
ng/
L;
the
downstream
concentrations
were
observed
to
increase
at
a
point
approximately
six
miles
below
the
outfall;
the
average
PFOS
concentration
from
that
point
downstream
was
114
+
19
ng/
L.
The
report
states
that
the
consistency
of
the
PFOS
concentrations
within
these
two
regions
suggests
the
absence
of
either
major
environmental
sinks
or
additional
sources
of
PFOS
in
the
areas
sampled.

The
3M
Environmental
Lab
reported
on
analyses
of
PFOS
in
naive
rat
livers,
rat
chow,
and
fish
meal
(
a
component
of
rat
chow).
Rat
livers
from
two
of
three
suppliers
had
endogenous
levels
of
PFOS
above
the
limit
of
detection
(
15
ng/
g).
PFOS
levels
in
the
male
rats
with
measurable
levels
increased
with
age,
while
the
levels
in
female
rats
showed
no
correlation
with
age.
In
the
rat
chow
study,
two
of
the
four
chows
examined
had
PFOS
above
the
2­
10
ng/
g
level
of
quantitation,
18
and
12
ng/
g,
respectively.
The
fish
meal
study
analyzed
PFOS
levels
in
six
types
of
fishmeal
formulated
from
at
least
three
types
of
fish.
Three
of
the
six
types
had
PFOS
levels
above
the
limit
of
quantitation
of
3.5
ng/
g;
the
maximum
concentration
was
15.7
ng/
g.

The
3M
Environmental
Laboratory
also
summarized
PFOS
analyses
of
avian
and
aquatic
feed
matrices.
PFOS
levels
were
not
detected
in
the
two
daphnid
feeds,
and
were
below
the
limits
of
quantitation
in
the
avian
feed.
Dry
artemia
(
brine
shrimp)
cysts
used
for
fish
feed
had
PFOS
levels
of
9­
10
ug/
kg.

2.4
Human
Biomonitoring
For
many
years,
PFOS
has
been
measured
in
the
serum
of
workers
occupationally
exposed
to
PFOS.
It
was
also
recently
detected
in
the
serum
of
the
general
population,
but
at
much
lower
levels.
The
results
of
the
most
recent
analyses
of
blood
samples
are
described
below
and
summarized
in
Table
1.

2.4.1
Occupational
Exposures
PFOS
serum
levels
have
been
measured
in
workers
involved
in
both
the
manufacturing
of
perfluorochemicals
and
the
processing
of
these
compounds
into
products,
such
as
fire
protection
and
surface
protection
products.
Biomonitoring
data
are
available
from
manufacturing
plants
in
Alabama
and
Belgium,
a
processing
facility
in
Japan,
and
corporate
offices
in
Minnesota.
ENV/
JM/
RD(
2002)
17/
FINAL
18
Fluorochemical
Manufacturing
Two
3M
plants,
located
in
Decatur,
Alabama
and
Antwerp,
Belgium,
have
produced
POSF
from
electrochemical
cell
fluorination
and
then
subsequently
used
POSF
to
produce
various
products
through
polymerization
processes.
These
POSF­
based
products
degrade
or
metabolize
to
PFOS.
A
third
manufacturing
plant,
located
in
Cottage
Grove,
Minnesota,
has
not
produced
POSF,
but
has
manufactured
some
POSF­
based
products.
Serum
PFOS
levels
were
measured
in
manufacturing
plant
employees
in
1995
(
n
=
178),
1997
(
n
=
149),
and
2000
(
n
=
521)
using
high
performance
liquid
chromatography/
mass
spectrometry
(
Olsen
et
al,
1999;
Olsen
et
al,
2001d;
Olsen
et
al,
2001e;
Olsen
et
al.,
2001f).
Employees
volunteered
to
participate
for
medical
surveillance.
Arithmetic
means
and
ranges
were
reported
for
all
surveillance
periods.
Geometric
means
have
only
been
reported
in
2000.
These
data
are
provided
in
Table
1.

Mean
serum
PFOS
levels
have
been
declining
at
both
plants
since
1995.
Decatur
serum
levels
have
been
higher
than
Antwerp
levels
during
every
surveillance
period.
The
PFOS
levels
in
workers
across
both
plants
in
1995
ranged
from
0.10
ppm
to
12.83
ppm.
The
range
of
PFOS
levels
of
workers
in
1997
was
0.10
­
9.93
ppm.
The
largest
number
of
employees
at
each
plant
participated
in
the
2000
medical
surveillance
program
(
521
out
of
840
employees).
The
mean
serum
PFOS
level
at
the
Decatur
plant
was
1.32
ppm
(
range
=
0.06
 
10.06
ppm).
Most
of
the
employees
at
both
facilities
had
PFOS
serum
levels
that
were
<
6
ppm
across
all
of
the
sampling
periods.

Except
for
the
year
2000,
the
participation
rate
of
the
total
number
of
potentially
exposed
employees
in
the
medical
monitoring
program
from
which
these
data
were
derived
was
not
provided.
It
was
reported
that
the
employees
were
offered
a
medical
monitoring
program,
but
it
is
not
clear
how
many
of
them
actually
volunteered
to
participate.
In
addition,
age
of
the
employees
and
duration
of
employment
were
not
reported
for
most
of
the
surveillance
years,
and
it
is
unclear
how
many
of
the
total
number
of
employees
worked
at
each
plant.
Therefore,
it
should
be
noted
that
these
data
only
provide
a
snapshot
of
the
serum
levels
of
participating
workers
during
each
sampling
period.

In
order
to
address
the
voluntary
nature
of
the
monitoring
program
at
the
3M
plants,
a
random
sample
of
employees
from
the
Decatur
plant
was
chosen
to
measure
seven
different
fluorochemicals
in
employees'
blood
(
Olsen
et
al.,
1999).
Limited
job
information,
years
worked,
and
building
location
were
collected
to
provide
a
better
understanding
of
the
distribution
of
fluorochemical
serum
levels
in
employees.
The
random
sample
consisted
of
187
chemical
plant
workers
(
126
randomly
chosen
workers
and
61
volunteers)
and
76
film
plant
workers
(
60
randomly
chosen
and
16
volunteers).
The
levels
in
these
employees,
who
were
sampled
in
late
1998,
were
similar
to
those
reported
above.
Mean
PFOS
levels
were
1.505
ppm
(
range,
0.091­
10.600)
for
the
random
sample,
1.259
ppm
for
the
volunteers,
and
1.424
ppm
for
all
participants.
Geometric
means
were
0.944
ppm
(
95%
CI
0.787­
1.126)
for
chemical
plant
employees
and
0.136
ppm
(
95%
CI
0.114­
0.161)
for
film
plant
employees.
When
compared
by
job
description,
workers
in
the
chemical
plant
had
higher
PFOS
levels
than
workers
in
the
film
plant.
PFOS
levels
in
males
were
positively,
although
modestly,
associated
with
number
of
years
worked
in
the
chemical
plant.

Fluorochemical
Processing
Biomonitoring
data
were
submitted
on
workers
in
Japan
processing
perfluorinated
chemicals
into
fire
protection
products
and
surface
products
(
Burris
et
al.,
1999).
In
1999,
PFOS,
PFOA
and
PFHS
(
perfluorohexanesulfonate)
were
measured
in
employees'
serum
at
the
Sagamihara
plant.
Serum
was
drawn
from
both
production
employees
(
n
=
32)
who
regularly
handle
fluorochemicals
and
management
employees
(
n
=
32)
who
are
not
regularly
exposed.
Serum
was
also
drawn
from
management
employees
(
n
=
30)
at
the
Head
Office
in
Tokyo.
The
highest
PFOS
level
in
32
production
employees
was
0.628
ppm,
with
an
arithmetic
mean
of
0.135
ppm.
The
highest
PFOS
level
in
either
of
the
management
groups
was
ENV/
JM/
RD(
2002)
17/
FINAL
19
0.0967
ppm,
while
the
average
level
was
approximately
0.04
ppm.
The
results
of
biomonitoring
performed
at
the
Sagamihara
plant
indicate
lower
levels
of
PFOS
than
at
either
of
the
manufacturing
facilities
cited
above.

Although
the
exposures
would
not
necessarily
be
considered
"
occupational"
for
either
group,
PFOS
levels
in
corporate
staff/
managers
at
a
3M
plant
in
St.
Paul,
Minnesota
were
similar
to
those
reported
in
the
corporate
managers
in
Japan
(
3M
Report,
1999).
Thirty­
one
employees,
none
of
whom
had
worked
in
fluorochemical
production
or
research
and
development,
were
sampled
in
1998.
All
of
the
participating
employees
had
measurable
levels
of
PFOS
in
their
serum.
The
mean
PFOS
level
was
0.047
ppm,
with
a
range
of
0.028
to
0.096
ppm.
In
this
group
of
3M
employees,
age
was
significantly
associated
with
increased
serum
PFOS.
No
other
data
on
these
workers
were
provided.

2.4.2
Non­
occupational
Exposures
PFOS
has
been
measured
in
the
serum
of
the
general
U.
S.
population
and
in
small
numbers
of
historical
samples
(
dating
as
far
back
as
1957)
from
sources
throughout
the
world.
In
1998,
PFOS
levels
in
pooled
serum
from
2
commercial
sources
in
the
U.
S.
ranged
from
43­
45
ppb
and
26
­
45
ppb
(
3M
Report,
1999).
There
were
approximately
500
donors
in
the
first
source
and
an
estimated
200
donors
in
the
second.
No
other
data,
such
as
geographic
location
or
age
of
the
donors,
were
provided.
3M
also
analyzed
thirty­
five
lots
of
individual
or
pooled
human
sera
samples
from
U.
S.
chemical
or
biological
supply
companies
in
1999
(
3M
Letter,
1999).
These
samples
yielded
an
average
of
35
ppb
PFOS,
ranging
from
5
to
85
ppb.

Sera
pooled
from
18
regional
blood
banks
in
various
geographic
regions
of
the
U.
S.
were
analyzed
for
PFOS
in
1998
(
3M
Report,
1999).
There
were
68
pools
and
an
estimated
340­
680
donors.
The
overall
mean
PFOS
serum
level
across
the
pools
was
29.7
ppb.
The
PFOS
levels
varied
quite
a
bit
depending
on
the
geographic
location
of
the
blood
bank.
The
range
of
the
levels
across
geographic
regions
was
9
to
56
ppb,
while
the
range
of
the
averages
was
14
to
52
ppb.

Pooled
sera
from
blood
banks
in
Belgium
(
6
pools),
the
Netherlands
(
5
pools),
and
Germany
(
6
pools)
also
have
been
analyzed
for
PFOS.
Each
pool
had
15­
20
donors.
The
Netherlands
had
the
highest
PFOS
levels
(
mean
=
53
ppb)
and
Belgium
had
the
lowest
(
mean
=
17
ppb).

Individual
blood
samples
from
3
different
age
populations
were
recently
analyzed
for
PFOS
and
other
fluorochemicals
using
high­
pressure
liquid
chromatography/
electrospray
tandem
mass
spectrometry
(
HPLC/
ESMSMS)
(
Olsen
et
al.,
2002a,
2002b,
2002c).
The
studies'
participants
included
adult
blood
donors,
an
elderly
population
participating
in
a
prospective
study
in
Seattle,
WA,
and
children
from
23
states
participating
in
a
clinical
trial.
Overall,
the
PFOS
geometric
means
were
similar
across
all
3
populations
(
34.9
ppb,
31.0
ppb,
and
37.5
ppb,
respectively).
The
geometric
means
and
95%
tolerance
limits
and
their
upper
bounds
were
comparable
across
all
3
studies.
However,
the
upper
ranges
for
the
children
and
adults
were
much
higher
than
for
the
elderly
population.
It
is
not
clear
whether
this
is
the
result
of
geographic
differences
in
PFOS
levels
or
some
other
factor.
It
should
be
noted
that
PFOS
and
PFOA
were
highly
correlated
in
all
three
studies
(
r
=
.63,
r
=
.70,
and
r
=
.75).
The
details
of
each
study
are
provided
below.

Serum
PFOS
levels
in
645
blood
donors
(
332
males,
313
females),
aged
20­
69
years,
were
obtained
from
6
American
Red
Cross
blood
banks
(
Olsen
et
al.,
2002a).
The
blood
banks
were
located
in:
Los
Angeles,
CA;
Minneapolis/
St.
Paul,
MN;
Charlotte,
NC;
Boston,
MA;
Portland,
OR,
and
Hagerstown,
MD.
Each
blood
bank
was
requested
to
provide
approximately
10
samples
per
10­
year
age
intervals
(
20­
29,
30­
39,
etc.)
for
each
sex.
The
only
demographic
factors
known
for
each
donor
were
age,
gender,
and
location.

The
geometric
mean
serum
PFOS
level
for
all
locations
and
gender
was
34.9
ppb
(
95%
CI,
33.3­
36.5
ppb).
ENV/
JM/
RD(
2002)
17/
FINAL
20
The
range
was
<
LLOQ
(
4.3
ppb)
to
1656
ppb.
Males
had
significantly
higher
(
p
<
.05)
geometric
mean
PFOS
levels
than
females.
The
geometric
mean
for
all
males
was
37.8
ppb
(
95%
CI,
35.5­
40.3)
and
was
31.3
ppb
for
all
females
(
95%
CI,
30.0
 
34.3).
Age
was
not
an
important
predictor
of
adult
serum
fluorochemical
concentrations.
When
stratified
by
geographic
location,
the
highest
geometric
mean
for
PFOS
was
in
the
samples
from
Charlotte,
NC
(
51.5
ppb,
range:
19.3
 
166.0)
and
the
lowest
from
Boston
(
28.0
ppb,
range:
4.3
 
87.2).

The
highest
serum
PFOS
measurement
in
this
sample
was
1656
ppb
from
a
male
blood
donor,
67
years
old
from
Portland.
The
next
highest
donor
level
was
329
ppb
from
a
male
donor,
62
years
old
also
from
Portland.
The
next
8
highest
serum
PFOS
values
(
range
139
 
226
ppb)
were
measured
in
4
females
and
4
males
representing
Charlotte
(
n=
4),
Hagerstown
(
n=
2),
Los
Angeles
(
n=
1)
and
Minneapolis/
St.
Paul
(
n=
1).
In
bootstrap
analyses,
the
mean
of
the
95%
tolerance
limit
for
PFOS
was
88.5
ppb
with
an
upper
95%
confidence
limit
of
100.0
ppb.

Serum
PFOS
levels
were
reported
for
238
(
118
males
and
120
females)
elderly
volunteers
in
Seattle
participating
in
a
study
designed
to
examine
cognitive
function
in
adults
aged
65­
96
(
Olsen
et
al.,
2002b).
Age,
gender
and
number
of
years'
residence
in
Seattle
were
the
only
data
available
on
the
participants.
Most
of
the
participants
were
under
the
age
of
85
and
had
lived
in
the
Seattle
area
for
over
50
years.

The
geometric
mean
of
PFOS
for
all
samples
was
31.0
ppb
(
95%
CI,
28.8­
33.4).
The
range
was
3.4
 
175
ppb.
There
was
no
significant
(
p
<
.05)
difference
in
geometric
means
for
males
and
females.
In
simple
linear
regression
analyses,
age
was
negatively
(
p
<
.05)
associated
with
PFOS
in
men
but
not
in
women.
The
mean
of
the
95%
tolerance
limit
for
PFOS
was
84.1
ppb
with
an
upper
95%
confidence
limit
of
104.0
ppb.

A
sample
of
599
children,
ages
2­
12
years
old,
participating
in
a
study
of
group
A
streptococcal
infections,
was
analyzed
for
PFOS
levels
(
Olsen
et
al.,
2002c).
The
samples
were
collected
in
1994­
1995
from
children
residing
in
23
states
and
the
District
of
Columbia.
The
geometric
mean
of
PFOS
for
all
of
the
participants
was
37.5
ppb
(
95%
CI,
33.3­
36.5).
The
range
was
6.7
to
515.0
ppb.
Male
children
had
significantly
(
p<.
01)
higher
geometric
mean
PFOS
levels
than
females:
40.1
ppb
and
35.2
ppb,
respectively.
In
bootstrap
analyses,
the
mean
of
the
95%
tolerance
limit
for
PFOS
was
88.5
ppb
with
an
upper
95%
confidence
limit
of
97.0
ppb.
When
stratified
by
age,
the
geometric
mean
tended
to
rise
for
each
age
group
from
age
2
(
28.6
ppb)
through
age
9
(
42.8
ppb)
where
it
was
highest,
and
then
started
to
decrease
gradually
to
32.8
ppb
at
12
years.
In
simple
linear
regression
analyses,
age
was
not
significantly
(
p
<
.05)
associated
with
PFOS.
Although
the
data
were
not
provided,
a
graphical
presentation
of
log
PFOS
levels
for
each
state
by
gender
were
similar
across
the
states,
however,
it
is
difficult
to
interpret
these
data
given
the
limited
sample
size
for
each
gender/
location
subgroup.

In
another
study,
Olsen
et
al.
(
2001g)
obtained
samples
from
31
cadavers
(
16
males
and
15
females)
over
an
18­
month
period
from
the
International
Institute
for
the
Advancement
of
Medicine
(
IIAM).
The
average
age
of
the
male
donors
was
50
years
(
SD
15.6,
range
5­
69)
and
the
average
age
of
the
female
donors
was
45
years
(
SD
18.5,
range
13­
74).
The
causes
of
death
were
intracranial
hemorrhage
(
n
=
16
or
52%),
motor
vehicle
accident
(
n
=
7
or
23%),
head
trauma
(
n
=
4
or
13%),
brain
tumor
(
n
=
2
or
6%),
drug
overdose
(
n
=
1
or
3%)
and
respiratory
arrest
(
n
=
1
or
3%).
Both
serum
and
liver
tissue
were
harvested
from
23
donors;
7
donors
contributed
liver
tissue
only
and
1
donor
contributed
serum
only.
Serum
samples
were
obtained
from
5
ml
of
blood;
liver
samples
consisted
of
10
g
of
tissue.
Samples
were
frozen
at
IIAM
and
shipped
frozen
to
3M
for
analysis.
The
samples
were
extracted
using
an
ion­
pairing
extraction
procedure
and
were
quantitatively
assayed
using
HPLC­
ESMSMS
and
evaluated
versus
an
unextracted
curve.
Extensive
matrix
spike
studies
were
performed
to
evaluate
the
precision
and
accuracy
of
the
extraction
procedure.
The
average
fortified
sample
recovery
of
PFOS
from
human
sera
was
89%
(
SD
21%)
and
the
average
fortified
sample
recovery
of
PFOS
from
human
liver
was
78%
(
SD
24%).
ENV/
JM/
RD(
2002)
17/
FINAL
21
Serum
PFOS
levels
that
were
determined
to
be
less
than
the
limit
of
quantitation
(
LOQ)
were
assigned
a
value
midpoint
between
zero
and
the
LOQ.
The
mean
serum
PFOS
level
for
the
24
serum
donors
analyzed
was
17.7
ng/
mL;
the
geometric
mean
for
serum
PFOS
was
14.7
ng/
mL.
Of
the
30
PFOS
liver
samples
analyzed,
15
(
50%)
were
determined
to
be
<
LOQ.
These
samples
were
assigned
a
value
midpoint
between
zero
and
the
LOQ.
The
mean
liver
PFOS
level
was
18.8
ng/
g;
the
geometric
mean
for
liver
PFOS
was
15.2
ng/
g.
Mean
PFOS
levels
for
male
and
female
donors
were
similar
for
both
serum
(
male
=
18.2
ng/
mL;
female
=
17.2
ng/
mL)
and
liver
(
male
=
19.2
ng/
g;
female
=
28.4
ng/
g).
Although
the
data
were
not
shown,
the
report
stated
that
no
associations
were
observed
between
measured
PFOS
levels
and
age.
The
average
PFOS
serum
and
liver
data
for
each
of
the
23
paired
samples
(
serum
and
liver
from
the
same
individual)
showed
a
good
correlation.
The
mean
liver
to
serum
ratio
was
1.3:
1.
Of
the
13
male
donors
with
paired
samples,
the
mean
liver
to
serum
ratio
was
again
1.3:
1;
the
mean
serum
level
was
18.2
ng/
mL
and
the
mean
liver
level
was
20.8
ng/
g.
For
the
10
female
donors
with
paired
samples,
the
mean
liver
to
serum
ratio
was
1.3:
1
with
mean
serum
levels
of
16.9
ng/
mL
and
mean
liver
levels
of
16.3
ng/
g,
respectively.
ENV/
JM/
RD(
2002)
17/
FINAL
22
Table
1.
Mean
PFOS
serum
levels
in
human
populations
Occupational
Exposures
Plant
Location
Mean
(
ppm)
Range
(
ppm)

Decatur,
Alabama
1995
(
n
=
90)
1997
(
n
=
84)
1998
(
n
=
126)
2000
(
n
=
263)
2.44
1.96
1.51
1.32
(
GM
=
0.91)*
0.25
­
12.83
0.10
­
9.93
0.09
 
10.6
0.06
 
10.06
(
33.3
 
36.5)**
Antwerp,
Belgium
1995
(
n
=
93)
1997
(
n
=
65)
2000
(
n
=
258)
1.93
1.48
0.80
(
GM
=
0.44)*
0.10
­
9.93
0.1
­
4.8
0.04
 
6.24
(
0.38
­
0.51)**
Building
236
(
n
=
45)
0.182
<
0.037
 
1.036
Sagamihara,
Japan
(
1999)
(
processing
PFOS)
n
=
32
0.135
0.0475
­
0.628
General
Population
Exposures
Source
Mean
(
ppb)
Range
(
ppb)

Non­
occupational
(
n
=
31)
(
corporate
staff
or
managers)
St.
Paul,
Minnesota
(
1998)
47
28
 
96
Non­
occupational
(
1999)
(
plant
management,
Japan)
Sagamihara
n
=
32
Tokyo
n=
30
40.3
52.3
31.9
­
56.6
33
­
96.7
Commercial
Sources,
U.
S.
(
1998)
Intergen
n
=
~
500
donors
Sigma
n
=
~
200
donors
44
33
43
­
44
26
 
45
Other
Commercial
Sources,
U.
S.
(
1999)
n
=
35
lots
35
5
 
85
U.
S.
Blood
Banks
(
1998)
n
=
~
340­
680
donors
29.7
9
 
56
European
Blood
Banks
(
1999)
Belgium
(
6
pooled
samples)

Netherlands
(
5
pooled
samples)

Germany
(
6
pooled
samples)
17
53
37
4.9
 
22.2
39
 
61
32
 
45.6
U.
S.
Blood
Banks
(
2000)
American
Red
Cross
(
ages
20­
69)
n
=
645
34.9
(
geometric
mean)
4.3
 
1656
Samples
in
U.
S.
children
(
ages
2­
12)
(
1995)
(
n
=
599)
37.5
(
geometric
mean)
6.7
 
515
Samples
in
elderly
in
Seattle,
WA
(
ages
65­
96)
(
1999)
(
n
=
238)
31.0
(
geometric
mean)
3.4
­
175
*
GM
is
geometric
mean;
**
95%
Confidence
Interval
of
the
GM
ENV/
JM/
RD(
2002)
17/
FINAL
23
3.0
Human
Health
Hazards
3.1
Metabolism
and
Pharmacokinetics
3.1.1
Absorption
PFOS
is
well
absorbed
following
ingestion.
After
a
single
oral
dose
of
PFOS­
14C
(
mean
dose,
4.2
mg/
kg)
in
solution
to
groups
of
three
male
rats,
at
least
95%
of
the
total
carbon­
14
is
systemically
absorbed
at
24
hours
(
Johnson,
Gibson
and
Ober,
1979a).
The
digestive
tract
and
contents
contained
on
average,
3.45%
of
the
dose.
The
mean
fecal
excretion
is
1.55%
of
the
dose
at
24
hours
and
3.24%
at
48
hours.
At
24
hours,
the
mean
sum
of
total
carbon­
14
in
feces
and
digestive
tract
plus
contents
is
5%
of
the
dose.
Some
of
this
5%
likely
represents
systemically
absorbed
carbon­
14
present
either
in
the
digestive
tract
tissues
or
in
the
digestive
tract
contents
as
a
result
of
excretion.
The
data
from
the
48
hour
post
dose
group
of
rats
are
consistent
with
the
24
hour
post
dose
data.
Thus,
at
least
95%
of
the
PFOS­
14C
dose
was
absorbed
from
solution
after
administration
to
non­
fasted
rats.

3.1.2
Distribution
At
24
to
48
hours
after
a
single
oral
dose
of
PFOS­
14C
(
4.2
mg/
kg)
in
rats,
approximately
86%
of
the
radioactivity
recovered
was
found
in
the
carcass
(
Johnson,
Gibson
and
Ober,
1979a).
The
carcass
data
are
not
as
reliable
as
the
other
tissue
data
since
large
volume
homogenates
were
necessary
and
homogeneity
of
sample
aliquots
was
difficult
to
assure.
There
is
some
excretion
of
total
carbon­
14
in
urine
(
1­
2%/
day).
The
spleens
from
the
24
hour
and
48
hour
post
dose
rats
were
analyzed
for
total
carbon­
14
content,
and
the
percent
of
the
dose
in
the
whole
organ
was
~
0.2%.
The
concentrations
of
total
carbon­
14
in
red
blood
cells
and
plasma
were
compared.
The
mean
ratio
of
red
blood
cell
to
plasma
concentration
at
24
and
48
hours
is
0.25
and
0.39,
respectively.
Thus,
at
24
and
48
hours
after
a
single
oral
dose
of
FC­
95­
14C,
there
is
no
selective
retention
of
carbon­
14
in
red
blood
cells.

At
89
days
after
a
single
intravenous
dose
of
PFOS­
14C
(
4.2
mg/
kg)
in
male
rats,
mean
tissue
C­
14
concentrations
above
one
ug
equivalents/
g
were
as
follows:
liver,
20.6;
plasma,
2.2;
kidney,
1.1;
and
lung,
1.1
(
Johnson,
Gibson
and
Ober,
1979b).
Other
tissues
such
as
muscle,
skin,
bone
marrow,
and
spleen
had
concentrations
ranging
from
0.2
to
0.6
ug/
g.
There
was
a
difference
in
C­
14
content
of
subcutaneous
fat
(
0.2
ug/
g)
and
abdominal
fat
(<=
0.08
ug/
g).
Very
little
C­
14
was
found
in
whole
eye
(
0.16
ug/
g)
and
no
detectable
C­
14
was
found
in
brain.
Only
liver
and
plasma
contained
a
substantial
percentage
of
dose
at
89
days
post
dose,
25.21%
and
2.81%,
respectively.
The
low
levels
of
radioactivity
found
for
kidney,
lung,
testes,
and
spleen
are
due
in
part
to
blood
still
contained
in
these
organs
when
homogenized.

There
have
been
several
studies
conducted
to
examine
the
levels
of
PFOS
in
the
dam,
fetus
and
neonate.
These
studies
are
described
in
detail
in
sections
3.4
and
3.5.
In
general,
the
levels
of
PFOS
are
higher
in
the
liver
than
in
the
serum
of
dams,
fetuses
and
neonates,
and
placental
transfer
of
PFOS
has
been
demonstrated
3.1.3
Elimination
Urinary
excretion
is
the
primary
route
of
elimination
for
PFOS
in
the
rat.
By
89
days
after
a
single
intravenous
dose
of
PFOS­
14C
(
4.2
mg/
kg)
in
male
rats,
mean
urinary
excretion
was
30.2+­
1.5%
of
total
C­
14
administered
(
Johnson,
Gibson,
and
Ober,
1979b).
Mean
cumulative
fecal
excretion
was
12.6+­
1.2%.
Radioactive
content
in
feces
was
too
low
to
measure
after
64
days.

The
half­
life
for
elimination
of
total
PFOS
carbon­
14
from
plasma
after
a
single
oral
dose
(
4.2
mg/
kg)
in
ENV/
JM/
RD(
2002)
17/
FINAL
24
male
rats
is
7.5
days
(
Johnson,
Gibson
and
Ober,
1979a).
This
determination
was
based
upon
analysis
of
plasma
samples
from
groups
of
three
rats
at
1,
2,
6,
12,
24,
48,
96,
and
144
hours
after
the
single
oral
dose.

There
is
evidence
of
enterohepatic
circulation
of
PFOS.
After
21
days
of
cholestyramine
treatment,
the
mean
percentage
of
PFOS­
14C
dose
eliminated
via
feces
(
75.8
+­
5.0)
was
9.5­
fold
the
mean
percentage
of
dose
eliminated
via
feces
by
control
rats
(
8.0
+­
0.8,
Johnson,
Gibson
and
Ober,
1984).
After
adjustment
for
the
amount
of
carbon­
14
excreted
in
urine
(
18%
for
controls
and
5%
for
cholestyramine­
treated),
the
amounts
of
carbon­
14
remaining
to
be
excreted
are
19%
for
cholestyramine­
treated
rats
and
74%
for
control
rats.
After
PFOS­
14C
administration,
the
mean
liver
carbon­
14
content
at
21
days
represents
11%
and
40%
of
the
dose
for
cholestyramine­
treated
and
control
rats,
respectively.
Mean
plasma
and
red
blood
cell
carbon­
14
concentrations
are
significantly
lower
after
21
days
of
cholestyramine
treatment.
The
authors
conclude
that
the
high
concentration
of
PFOS­
14C
in
liver
at
2
to
3
weeks
after
dosing
and
the
fact
that
cholestyramine
treatment
enhances
fecal
elimination
of
carbon­
14
by
nearly
10­
fold
suggest
that
there
is
a
considerable
enterohepatic
circulation
of
PFOS­
14C.

3.1.4
Half­
life
in
Humans
In
order
to
determine
the
half­
life
of
PFOS,
3
retirees
who
worked
in
3Ms
Chemical
Division
were
followed
for
five
and
a
half
years
(
3M
Company,
2000c).
Serum
PFOS
levels
suggested
a
mean
elimination
half­
life
(
t1/
2)
of
1,428
days
(
approximately
4
years).
A
half­
life
study
on
a
larger
group
of
retirees
(
n
=
27)
is
currently
underway,
in
which
serum
samples
are
drawn
every
6
months
over
a
5­
year
period.
Two
interim
reports
describing
the
results
thus
far
have
been
submitted
(
Burris
et
al,
2000;
Burris
et
al.,
2002).
The
first
interim
report
suggested
a
serum
half­
life
of
139­
640
days.
However,
there
were
several
limitations
to
these
analyses
including:
1)
the
limited
data
available
and
the
range
of
serum
PFOS
levels
measured;
2)
serum
was
analyzed
after
each
collection
period
with
only
one
measurement
per
time
period
on
different
days
using
slightly
different
analytical
techniques;
and
3)
the
reference
material
purity
was
not
determined
until
after
the
first
3
samples
had
been
analyzed.
An
effort
was
made
to
minimize
experimental
error,
including
systematic
and
random
error
in
the
analytical
method,
involving
9
of
the
original
27
subjects.
Serum
samples
were
collected
from
each
of
the
subjects
over
4
time
periods
spanning
180
days,
measured
in
triplicate
with
all
time
points
from
each
subject
analyzed
in
the
same
analytical
run.
This
would
allow
for
statistical
evaluation
of
the
precision
of
the
measurement
and
assure
that
all
systematic
error
inherent
in
the
assay
equally
affected
each
sample
used
for
half­
life
determination.

Of
the
9
retirees
included
in
this
analysis,
there
were
7
males
and
2
females,
all
from
the
Decatur
plant.
The
average
age
of
the
retirees
was
61
years,
the
mean
number
of
years
worked
at
Decatur
was
27.7
years,
and
the
average
number
of
months
retired
was
18.9.
Average
BMI
of
this
group
was
27.9.
The
mean
PFOS
value
at
study
initiation
was
0.89
ppm
(
range
0.11
 
3.53
ppm,
SD
=
1.07).

The
mean
serum
half­
life
for
PFOS
was
8.67
years
(
range
2.29
 
21.3
years,
SD
=
6.12).
Age,
BMI,
number
of
years
worked
or
years
since
retirement
were
not
significant
predictors
of
serum
half­
lives
in
multivariable
regression
analyses.
This
analysis
has
attempted
to
reduce
experimental
error
in
the
determination
of
a
half­
life
for
PFOS.
However,
several
issues
should
be
noted.
First,
the
effect
of
continued
non­
occupational,
low­
level
exposure
on
the
half­
life
is
unknown.
Second,
PFOS
is
a
metabolic
product
of
other
compounds
found
in
the
retirees'
blood;
therefore,
PFOS
is
likely
being
produced
in
the
body
while
the
study
was
ongoing.
Third,
it
is
not
known
if
there
are
interactions
between
PFOS
and
other
fluorochemicals
in
the
body.
Fourth,
this
estimate
is
much
higher
than
that
reported
in
lab
animals.
However,
it
may
not
be
appropriate
to
directly
compare
the
results
of
the
animal
half­
life
data
with
these
data
due
to
potentially
different
protein
binding
sites
and
affinities.
Fifth,
systematic
error
of
the
analytical
method
could
be
as
high
as
+/­
20%
and
still
satisfy
the
data
quality
criteria.
ENV/
JM/
RD(
2002)
17/
FINAL
25
3.2
Acute
Toxicity
Four
reports
of
acute
studies
of
PFOS
have
been
submitted,
one
inhalation
toxicity
of
rats,
two
oral
studies
of
rats,
and
one
dermal
and
eye
irritation
study
of
rabbits.

In
a
study
to
determine
the
median
lethal
concentration
(
LC50),
Rusch
et
al.
(
1979)
administered
PFOS
dust
in
air
to
Sprague­
Dawley
rats,
5/
sex/
group,
levels
of
1.89
to
45.97
mg/
l
PFOS
to
eight
test
groups.
A
Wright
dust­
feed
mechanism
with
dry
air
at
a
flow
rate
of
12
to
16
liters
per
minute
was
used
to
administer
the
PFOS
dust.
Rats
were
exposed
for
1
hour.
The
test
group
rats
weighed
201­
299
g
at
study
initiation.
The
control
group
rats
weighed
203­
263
g
at
study
initiation.
The
test
group
rats
were
exposed
to
1.89,
2.86,
4.88,
6.49,
7.05,
13.9,
24.09
or
45.97
g/
l.
The
control
rats
were
exposed
to
dry
air
at
a
flow
rate
of
12
liters
per
minute.
All
other
protocols
were
the
same
as
the
test
group
rats.
The
rats
were
observed
for
abnormal
signs
prior
to
exposure,
at
15­
minute
intervals
during
the
1­
hour
exposure,
at
removal
from
the
exposure
chamber,
hourly
for
four
hours
after
exposure,
and
daily
thereafter
for
14
days.
Individual
body
weights
were
recorded
on
Day
0
(
prior
to
exposure),
Day
1,
Day
2,
Day
4,
Day
7,
and
Day
14.
It
is
reported
that
all
animals
dying
spontaneously
were
necropsied
as
soon
as
possible
after
death.
Blood
samples
were
collected
on
Day
14
from
all
surviving
animals,
but
analyses
were
not
provided.
The
LC50
of
5.2
mg/
l
and
95%
confidence
limits
of
4.4
and
6.4
mg/
l
were
calculated
using
the
method
of
Litchfield
and
Wilcoxon.

The
highest
dose
group,
45.97
mg/
l,
was
not
used
in
the
LC50
calculations
and
terminated
on
Day
2.
At
that
point,
only
5
animals
survived
and
blood
samples
were
taken
at
termination.
The
13.9
mg/
l
group
was
also
terminated
early
(
Day
1)
because
of
a
mechanical
problem
during
exposure.
These
animals
were
also
not
used
in
the
LC50
determination.

In
the
24.09
mg/
l
exposure
group,
all
animals
died
by
Day
6.
At
7.05
and
6.49
mg/
l
there
was
80%
mortality.
At
4.88,
2.86,
and
1.89
mg/
l
there
was
20%,
10%,
and
0%
mortality,
respectively.
The
rats
in
all
these
groups
showed
signs
of
toxicity
including
emaciation,
red
material
around
the
nose
or
other
nasal
discharge,
yellow
material
around
the
anogenital
region,
dry
rales
or
other
breathing
disturbances,
and
general
poor
condition.
Abnormal
in­
life
observations
were
reported
to
be
less
frequent
in
the
lower
exposure
groups.

At
necropsy,
the
most
common
abnormality
was
discoloration
of
the
liver
and
lung.
Discoloration
of
the
lung
was
also
observed
in
control
rats
and
therefore
may
not
be
treatment
related.
Therefore,
the
most
significant
treatment
related
abnormality
was
varying
degrees
of
discoloration
of
the
liver.
Among
animals
that
died
prematurely,
decreased
body
weight,
discoloration
of
the
lung,
and
discoloration
and
distention
of
the
small
intestine
were
also
observed.

In
a
study
to
determine
the
acute
oral
LD50,
Dean
et
al.
(
1978)
administered
CD
rats,
5/
sex/
group,
doses
of
0,
100,
215,
464,
and
1000
mg/
kg
PFOS
by
gavage.
The
powdered
test
material
was
suspended
in
a
20%
acetone/
80%
corn
oil
mixture.
All
dose
levels
were
administered
as
volumes
of
10
ml/
kg
body
weight.
The
rats
weighed
172­
212
g
at
the
beginning
of
the
study
immediately
prior
to
dosing
and
weights
were
recorded
at
Day
7
and
Day
14.
The
rats
were
observed
for
abnormal
signs
during
the
four
hours
after
exposure,
and
daily
thereafter
for
14
days.
It
is
reported
that
all
animals
dying
spontaneously
were
grossly
necropsied,
as
well
as
all
rats
that
survived
to
the
end
of
the
14
day
study.

Acute
oral
LD50
values
and
95%
confidence
limits
were
calculated
for
males
(
233
[
160­
339]
mg/
kg),
females
(
271
[
200­
369]
mg/
kg)
and
combined
male
and
female
rats
(
251
[
199­
318]
mg/
kg).

All
rats
in
the
464
and
1000
mg/
kg
dose
groups
died
before
the
end
of
the
study.
Three
animals
in
the
215
mg/
kg
group
died
prematurely.
It
appears
signs
of
toxicity
most
frequently
observed
included:
ENV/
JM/
RD(
2002)
17/
FINAL
26
hypoactivity,
decreased
limb
tone,
and
ataxia.
At
necropsy,
observations
included:
yellow­
stained
urogenital
region,
stomach
distention
and
signs
of
irritation
of
the
glandular
mucosa,
and
lung
congestion.

In
a
second
oral
study,
Gabriel
(
1978;
cited
in
Dean
et
al.,
1978)
administered
two
groups
of
Sherman­
Wistar
albino
rats
(
5
/
sex/
group)
two
doses
of
PFOS
by
gavage
in
water.
The
rats
weighed
between
200
­
300
grams.
It
was
determined
the
acute
oral
LD50
was
greater
than
50
mg/
kg
and
less
than
1500
mg/
kg.

All
of
the
rats
administered
1500
mg/
kg
died
before
the
14­
day
observation
period
ended,
with
the
last
rat
dead
on
Day
9.
None
of
the
rats
administered
50
mg/
kg
died
before
the
end
of
the
study.
It
is
stated
that
rats
were
observed
for
signs
of
toxicity
and
mortality
but
it
is
not
clear
how
frequently
they
were
observed
and
no
individual
animal
data
was
provided.
It
is
stated
that
the
high
dose
group
rats
were
"
depressed"
at
2­
3
hours
after
dosing
and
"
severely
depressed
or
semi­
comatose"
by
24
hours;
staying
in
this
state
until
death
occurred.
No
individual
pathology
data
were
provided.
The
author
states:
"
Gross
pathologic
examination
revealed
nothing
remarkable."

Biesemeier
and
Harris
(
1974)
examined
the
potential
for
PFOS
to
cause
skin
and
eye
irritation.
Six
albino
rabbits
had
their
hair
clipped
from
their
backs
and
flanks,
and
it
appears
five
tenths
of
one
gram
(
0.5
g;
the
procedure
states
either
0.5
g
or
0.5
ml
and
the
test
material
as
supplied
by
3M
was
a
solid)
of
the
test
material
was
placed
on
abraded
or
intact
prepared
test
sites,
then
covered
with
gauze
patches.
After
24
hours
and
72
hours
the
coverings
were
removed
and
the
degree
of
erythema
and
edema
was
recorded
according
to
a
standardized
scale.
No
reference
is
provided
for
method
or
scoring
of
results.
Concentration
or
total
dose
of
test
material
was
not
provided.
In
all
six
rabbits,
it
is
reported
the
primary
skin
irritation
scores
were
0;
which
indicates
no
reddening
or
swelling
detected.

In
the
eye
irritation
study,
six
albino
New
Zealand
White
rabbits
were
placed
in
collars
so
they
could
not
rub
their
eyes.
It
appears
one
tenth
of
a
gram
(
0.1
g;
the
procedure
states
either
0.1
g
or
0.1
ml
was
used
and
T117
was
a
solid
as
supplied
by
3M)
of
the
test
substance
was
instilled
in
one
eye,
the
other
eye
was
left
untreated
as
a
control.
The
concentration
or
total
dose
of
the
test
substance
were
not
provided.
It
is
reported
that
the
reaction
to
the
test
material
was
read
against
a
scale
of
damage
to
the
cornea,
iris,
and
the
bulbar
and
palpebral
conjunctivae
at
1,
24,
48,
and
72
hours
after
treatment.
The
scale
criteria
were
not
presented
or
referenced.
Each
time
the
eyes
were
scored,
any
accumulated
discharge
or
residue
of
test
material
was
flushed
from
the
eye.
There
is
no
reference
provided
for
the
method.
It
appears
that
scores
were
maximal
at
1
hour
and
24
hours
after
treatment
then
decreased
over
the
rest
of
the
study
(
Biesemeier
and
Harris,
1974).

3.3
Mutagenicity
PFOS
was
tested
for
its
ability
to
induce
mutation
in
the
Ames
Salmonella/
Microsome
Plate
Test
and
in
the
D4
strain
of
Saccharomyces
cerevisiae
(
Litton
Bionetics,
Inc.,
1979).
It
was
also
tested
in:
(
1)
a
Salmonella
 
Escherichia
coli/
Mammalian­
microsome
reverse
mutation
assay
(
Mecchi,
1999);
(
2)
an
in
vitro
assay
for
chromosomal
aberrations
in
human
whole
blood
lymphocytes
(
Murli,
1999);
and
(
3)
an
assay
for
unscheduled
DNA
synthesis
(
UDS)
in
rat
liver
primary
cell
cultures
(
Cifone,
1999).
It
was
negative
in
all
assays
in
which
it
was
tested.
PFOS
does
not
induce
reverse
mutation
at
the
histidine
locus
of
S.
typhimurium
or
at
the
tryptophan
locus
of
E.
coli
when
tested
with
or
without
metabolic
activation.
It
does
not
induce
chromosomal
aberrations
in
human
lymphocytes
when
tested
with
or
without
metabolic
activation
and
does
not
induce
UDS
in
primary
cultures
of
rat
hepatocytes.
PFOS
was
also
tested
in
an
in
vivo
mouse
micronucleus
assay
(
Murli,
1996).
PFOS
did
not
induce
micronuclei
in
the
bone
marrow
of
Crl:
CD­
1
BR
mice
and
is
negative
in
the
mouse
bone
marrow
micronucleus
assay.

T­
2247
CoC
which
is
a
50%
by
weight
solution
of
the
diethanolammonium
salt
of
perfluorooctanesulfonate
in
water
was
tested
for
its
ability
to
induce
gene
mutation
in
5
strains
of
S.
ENV/
JM/
RD(
2002)
17/
FINAL
27
typhimurium
and
also
for
its
ability
to
induce
recombination
in
S.
cerevisiae
strain
D3.
(
Simmon,
1978).
The
chemical
was
negative
in
all
5
strains
of
Salmonella
and
in
S.
Cerevisiae
D3
when
tested
with
and
without
metabolic
activation.
T­
2247
CoC
was
tested
with
S.
typhimurium
strains
TA98
and
TA100
in
a
desiccator
assay
for
the
detection
of
volatile
compounds.
It
was
nonmutagenic
when
tested
under
these
conditions.

3.4
Repeated
Dose
Toxicity
Three
90­
day
subchronic
studies
of
PFOS
have
been
conducted,
two
gavage
studies
in
rhesus
monkeys
and
one
dietary
study
in
rats.
In
addition,
a
four
week
and
a
26
week
capsule
study
in
cynomolgus
monkeys
have
been
conducted.
Both
reports
were
unaudited
drafts
at
the
time
of
this
review.

In
the
rat
subchronic
study,
Goldenthal
et
al.
(
1978b)
administered
CD
rats,
5/
sex/
group,
dietary
levels
of
0,
30,
100,
300,
1000
or
3000
ppm
PFOS
(
FC­
95)
for
90
days.
The
males
weighed
196­
232
g
and
the
females
weighed
165­
206
g
at
study
initiation.
The
dietary
levels
were
equivalent
to
doses
of
0,
2,
6,
18,
60
and
200
mg/
kg/
day.
The
rats
were
observed
daily
for
general
clinical
signs;
body
weights
and
food
consumption
were
recorded
weekly.
Hematological
and
clinical
chemistry
analyses
and
urinalysis
were
conducted
at
the
beginning
of
the
study
and
after
30
and
90
days
of
treatment.
The
rats
were
sacrificed
after
90
days
of
treatment
and
a
gross
necrospy
was
conducted.
At
necropsy
the
heart,
liver,
adrenals,
spleen,
pituitary,
kidneys,
testes/
ovaries
and
brain
were
weighed.
The
thyroid/
parathyroid
were
weighed
after
fixation.
Tissues
were
preserved
in
buffered
neutral
10%
formalin;
the
eyes
were
preserved
in
Russell's
fixative.
The
following
organs
from
control
and
all
treated
groups
were
examined
microscopically:
adrenals,
aorta,
brain,
esophagus,
eyes,
gallbladder,
heart
(
with
coronary
vessels),
duodenum,
ileum,
jejunum,
cecum,
colon,
rectum,
kidneys,
liver,
lung,
skin,
mesenteric
lymph
node,
retropharyngeal
lymph
node,
mammary
gland,
nerve
(
with
muscle),
spleen,
pancreas,
prostate/
uterus,
bone/
bone
marrow
(
rib
junction),
salivary
gland,
lumbar
spinal
cord,
pituitary,
stomach,
testes/
ovaries,
thyroid,
parathyroid,
thymus,
trachea,
tonsil,
tongue,
urinary
bladder
and
vagina.

All
of
the
rats
in
the
300,
1000
and
3000
ppm
groups
died.
Death
occurred
between
days
13­
25
and
days
18­
28
for
the
males
and
females,
respectively,
in
the
300
ppm
group.
At
1000
ppm,
death
occurred
between
days
8­
14,
and
at
3000
ppm,
the
rats
died
between
days
7­
8
of
treatment.
The
rats
in
all
groups
showed
signs
of
toxicity
including
emaciation,
convulsions
following
handling,
hunched
back,
red
material
around
the
eyes,
yellow
material
around
the
anogenital
region,
increased
sensitivity
to
external
stimuli,
reduced
activity
and
moist
red
material
around
the
mouth
or
nose.

Three
males
and
two
females
in
the
100
ppm
group
died
prior
to
scheduled
sacrifice.
Two
of
the
males
and
the
two
females
died
during
week
5
and
the
third
male
died
during
week
11
of
the
study.
At
study
termination,
mean
body
weights
were
reduced
by
16.7%
and
16.3%
in
the
male
and
female
groups,
respectively.
Average
food
consumption
during
the
entire
study
period
(
g/
rat/
day)
was
significantly
reduced
for
males
and
females
at
100
ppm.
After
30
days
of
treatment,
hematologic
values
were
comparable
among
the
control
and
100
ppm
groups.
Clinical
chemistry
analyses
at
one
month
showed
a
significant
increase
in
mean
glucose
in
males,
blood
urea
nitrogen
values
in
males
and
females,
and
creatinine
phosphokinase
and
alkaline
phosphatase
values
for
females.
After
90
days
of
treatment
at
100
ppm,
the
two
surviving
males
had
significantly
reduced
erythrocyte,
hemoglobin,
hematocrit
and
leukocyte
counts;
the
three
surviving
females
had
significantly
reduced
hemoglobin
and
reticulocyte
counts,
as
well
as
slightly
lower
erythrocyte,
hematocrit
and
leukocyte
counts.
Two
of
the
surviving
females
showed
slight
to
moderate
increases
in
plasma
glutamic
oxalacetic
and
pyruvic
transaminase
activities.
Urinalysis
results
were
comparable
among
treated
and
control
groups
at
30
and
90
days.
Relative
liver
weight
was
significantly
increased
in
the
males
and
absolute
and
relative
liver
weights
were
significantly
increased
in
the
females.
Relative
kidney
weights
were
significantly
increased
in
both
sexes.
ENV/
JM/
RD(
2002)
17/
FINAL
28
All
rats
in
the
30
ppm
group
survived
until
the
end
of
the
study.
At
study
termination,
mean
body
weights
were
reduced
by
8.7
and
8%
in
the
males
and
females,
respectively.
Average
food
consumption
during
the
entire
study
period
(
g/
rat/
day)
was
significantly
reduced
for
the
males
at
30
ppm.
Hematologic
values
were
comparable
among
the
control
and
30
ppm
group
at
30
and
90
days.
One
female
showed
a
slightly
elevated
glucose
level
and
one
male
showed
a
slightly
increased
alkaline
phosphatase
level
at
30
days.
At
90
days,
one
male
showed
moderate
increases
in
glucose,
blood
urea
nitrogen
and
y­
glutamyl
transpeptidase
activity.
The
females
had
significant
increases
in
absolute
and
relative
liver
weights.
The
males
had
significant
decreases
in
absolute
and
relative
adrenal
weights,
absolute
thyroid/
parathyroid
weight
and
absolute
pituitary
weight.
The
biological
significance
of
the
changes
in
male
organ
weights
is
unclear
since
similar
changes
were
not
noted
in
higher
dose
groups.

At
necropsy,
treatment
related
gross
lesions
were
present
in
all
treated
groups
and
included
varying
degrees
of
discoloration
and/
or
enlargement
of
the
liver
and
discoloration
of
the
glandular
mucosa
of
the
stomach.
Histologic
examination
also
showed
lesions
in
all
treated
groups.
Centrilobular
to
midzonal
cytoplasmic
hypertrophy
of
hepatocytes
and
focal
necrosis
was
observed
in
the
liver;
the
incidence
and
relative
severity
were
greater
in
the
males.
In
addition,
especially
among
rats
in
the
300,
1000
and
3000
ppm
groups,
treatment
related
histologic
lesions
were
noted
in
the
primary
(
thymus,
bone
marrow)
and
secondary
(
spleen,
mesenteric
lymph
nodes)
lymphoid
organs,
stomach,
intestines,
muscle
and
skin.
In
the
thymus,
this
consisted
of
depletion
in
the
number
and
size
of
the
lymphoid
follicles
and
in
the
bone
marrow
hypocellularity
was
noted.
The
spleen
was
slightly
atrophied
with
a
corresponding
decrease
in
the
size
and
number
of
lymphoid
follicles
and
cells
and
a
similar
depletion
was
noted
in
the
mesenteric
lymph
nodes.
Mucosal
hyperkeratosis
and/
or
acanthosis
was
observed
in
the
forestomach
and
mucosal
hemorrhages
were
noted
in
the
glandular
portion
of
the
stomach.
Decreases
in
the
height
and
thickness
of
the
villi
were
noted
in
the
small
intestine.
Atrophy
of
the
skeletal
muscle
was
noted,
as
well
as
epidermal
hyperkeratosis
and/
or
acanthosis
was
noted
in
the
skin.

In
the
first
rhesus
monkey
study,
Goldenthal
et
al.
(
1979)
administered
rhesus
monkeys,
2/
sex/
group,
doses
of
0,
10,
30,
100
or
300
mg/
kg/
day
PFOS
(
FC­
95)
in
distilled
water
by
gavage.
The
males
weighed
3.05­
3.80
kg
at
study
initiation
and
the
females
weighed
2.75­
4.10
kg.
The
monkeys
were
observed
daily
for
general
clinical
signs
and
body
weights
were
recorded
weekly.
Hematological
and
clinical
chemistry
analyses
and
urinalysis
were
conducted
at
the
beginning
of
the
study.
The
study
was
terminated
after
20
days
due
to
the
death
of
the
monkeys.
At
necropsy
the
heart,
liver,
adrenals,
spleen,
pituitary,
kidneys,
testes/
ovaries
and
brain
were
weighed.
The
thyroid/
parathyroid
were
weighed
after
fixation.
Tissues
were
preserved
in
buffered
neutral
10%
formalin;
the
eyes
were
preserved
in
Russell's
fixative.
The
following
organs
from
control
and
all
treated
groups
were
examined
microscopically:
adrenals,
aorta,
brain,
esophagus,
eyes,
gallbladder,
heart
(
with
coronary
vessels),
duodenum,
ileum,
jejunum,
cecum,
colon,
rectum,
kidneys,
liver,
lung,
skin,
mesenteric
lymph
node,
retropharyngeal
lymph
node,
mammary
gland,
nerve
(
with
muscle),
spleen,
pancreas,
prostate/
uterus,
bone/
bone
marrow
(
rib
junction),
salivary
gland,
lumbar
spinal
cord,
pituitary,
stomach,
testes/
ovaries,
thyroid,
parathyroid,
thymus,
trachea,
tonsil,
tongue,
urinary
bladder
and
vagina.

All
of
the
treated
monkeys
died.
The
monkeys
in
the
300
mg/
kg/
day
group
died
between
days
2­
4,
the
monkeys
in
the
100
mg/
kg/
day
group
died
between
days
3­
5,
the
monkeys
in
the
30
mg/
kg/
day
group
died
between
days
7­
10,
and
the
monkeys
in
the
10
mg/
kg/
day
group
died
between
days
11­
20
of
treatment.
The
monkeys
from
all
the
groups
showed
similar
signs
of
toxicity
including
decreased
activity,
emesis
with
some
diarrhea,
body
stiffening,
general
body
trembling,
twitching,
weakness,
convulsions
and
prostration.
At
necropsy,
several
of
the
monkeys
in
the
100
and
300
mg/
kg/
day
groups
had
a
yellowishbrown
discoloration
of
the
liver;
histologic
examination
showed
no
microscopic
lesions.
Congestion,
hemorrhage
and
lipid
depletion
of
the
adrenal
cortex
was
noted
in
all
treated
groups.
No
other
lesions
were
noted.
ENV/
JM/
RD(
2002)
17/
FINAL
29
In
the
second
study,
Goldenthal
et
al.
(
1978a)
administered
rhesus
monkeys,
2/
sex/
group,
doses
of
0,
0.5,
1.5
or
4.5
mg/
kg/
day
PFOS
(
FC­
95)
in
distilled
water
by
gavage
for
90
days.
The
males
weighed
2.55­
3.55
kg
at
study
initiation
and
the
females
weighed
2.7­
3.75
kg.
The
monkeys
were
observed
daily
for
general
clinical
signs
and
body
weights
were
recorded
weekly.
Hematological
and
clinical
chemistry
analyses
and
urinalysis
were
conducted
at
the
beginning
of
the
study
and
after
30
and
90
days
of
treatment.
The
monkeys
were
sacrificed
after
90
days
of
treatment
and
a
gross
necrospy
was
conducted.
At
necropsy
the
heart,
liver,
adrenals,
spleen,
pituitary,
kidneys,
testes/
ovaries
and
brain
were
weighed.
The
thyroid/
parathyroid
were
weighed
after
fixation.
Tissues
were
preserved
in
buffered
neutral
10%
formalin;
the
eyes
were
preserved
in
Russell's
fixative.
The
following
organs
from
control
and
all
treated
groups
were
examined
microscopically:
adrenals,
aorta,
brain,
esophagus,
eyes,
gallbladder,
heart
(
with
coronary
vessels),
duodenum,
ileum,
jejunum,
cecum,
colon,
rectum,
kidneys,
liver,
lung,
skin,
mesenteric
lymph
node,
retropharyngeal
lymph
node,
mammary
gland,
nerve
(
with
muscle),
spleen,
pancreas,
prostate/
uterus,
bone/
bone
marrow
(
rib
junction),
salivary
gland,
lumbar
spinal
cord,
pituitary,
stomach,
testes/
ovaries,
thyroid,
parathyroid,
thymus,
trachea,
tonsil,
tongue,
urinary
bladder
and
vagina.

All
monkeys
in
the
4.5
mg/
kg/
day
group
died
or
were
sacrificed
in
extremis
between
week
5
and
7
of
the
study.
Beginning
on
the
first
or
second
day
of
the
study,
these
monkeys
exhibited
signs
of
gastrointestinal
tract
toxicity
including
anorexia,
emesis,
black
stool
and
dehydration.
All
of
the
monkeys
had
decreased
activity
and
just
prior
to
death
showed
marked
to
severe
rigidity,
convulsions,
generalized
body
trembling
and
prostration.
The
mean
body
weight
decreased
from
3.44
kg
at
the
beginning
of
the
study
to
2.7
kg
at
week
5.
After
30
days
of
treatment,
there
was
a
significant
reduction
in
serum
cholesterol
and
a
50%
reduction
in
serum
alkaline
phosphatase
activity.
At
necropsy,
mean
organ
weights
were
comparable
among
the
control
and
treated
monkeys.
Histologic
examination
showed
several
treatment
related
lesions.
All
the
male
and
females
had
marked
diffuse
lipid
depletion
in
the
adrenals.
One
male
and
two
females
had
moderate
diffuse
atrophy
of
the
pancreatic
exocrine
cells
with
decreased
cell
size
and
loss
of
zymogen
granules.
Two
males
and
one
female
had
moderate
diffuse
atrophy
of
the
serous
alveolar
cells
characterized
by
decreased
cell
size
and
loss
of
cytoplasmic
granules.

All
monkeys
in
the
1.5
mg/
kg/
day
group
survived
until
the
end
of
the
study.
During
the
first
week
of
the
study,
the
monkeys
had
decreased
activity.
Signs
of
gastrointestinal
tract
toxicity
were
noted
occasionally
during
the
study
and
included
black
stool,
diarrhea,
mucous
in
the
stool
and
bloody
stool;
at
the
end
of
the
study,
anorexia,
dehydration
or
general
body
trembling
were
noted.
Although
statistical
significance
was
not
achieved,
the
mean
body
weight
of
the
males
dropped
from
3.15
kg
at
the
beginning
of
the
study
to
2.93
kg
at
the
end
of
the
study,
and
the
mean
body
weight
of
the
females
dropped
from
3.22
kg
to
2.75
kg.
One
of
the
females
had
very
low
serum
cholesterol
and
another
had
a
reduction
in
inorganic
phosphate.
Necropsy
revealed
no
treatment
related
lesions.

All
monkeys
in
the
0.5
mg/
kg/
day
group
survived
until
the
end
of
the
study.
Signs
of
gastrointestinal
tract
toxicity
were
noted
occasionally
during
the
study
and
included
diarrhea,
soft
stools,
anorexia
and
emesis.
Occasionally,
decreased
activity
was
noted
in
three
of
the
monkeys.
Necropsy
revealed
no
treatment
related
lesions.

Serum
samples
from
monkeys
in
the
0,
0.5
and
1.5
mg/
kg/
day
dose
group
were
individually
analyzed
for
PFOS
levels;
liver
samples
from
the
0,
0.5,
1.5
and
4.5
mg/
kg/
day
dose
groups
were
also
analyzed
for
PFOS
levels.
Except
for
the
control
animals
where
animal
7355M
was
used
in
both
the
serum
and
liver
control
groups,
different
animals
were
used
for
serum
and
liver
determinations.
PFOS
levels
in
serum
were
developed
by
pyrolysis;
precision
is
estimated
to
be
±
10
 
25%.
ENV/
JM/
RD(
2002)
17/
FINAL
30
Levels
of
PFOS
in
serum
Monkey
#
Dosage
Group
PFOS
Levels
in
Serum
(
ppm)
7355M
0
40
7358M
0
20
7368F
0
15
7460M
0.5
mg/
kg/
day
150
7466F
0.5
mg/
kg/
day
150
7462M
1.5
mg/
kg/
day
250
7500F
1.5
mg/
kg/
day
275
No
explanation
is
offered
for
the
relatively
high
serum
levels
in
the
controls.
Sample
7355M
was
run
on
two
separate
days
and
gave
the
same
results.

Levels
of
PFOS
in
liver
Monkey
#
Dosage
Level
PFOS
in
Liver
(
mg)
PFOS
in
Liver
(
ppm)
7355M
0
3000
50
7368F
0
1500
20
7463M
0.5
mg/
kg/
day
7000
100
7466F
0.5
mg/
kg/
day
8000
100
7462M
1.5
mg/
kg/
day
45000
650
7500F
1.5
mg/
kg/
day
40000
600
7484M
4.5
mg/
kg/
day
40000
650
7502F
4.5
mg/
kg/
day
80000
1000
Livers
were
stored
refrigerated
for
several
months
prior
to
analysis.
This
resulted
in
the
separation
of
some
liquid
from
the
samples.
At
the
time
of
analysis,
only
partial
samples
were
taken
in
which
the
ratio
of
solid
to
liquid
was
estimated
visually.
Recovery
of
PFOS
from
the
liver
samples
was
estimated
from
experiments
where
a
known
amount
of
PFOS
was
added
to
a
control
liver.
Here
as
in
the
serum,
control
values
were
higher
than
expected.
Because
of
this,
a
0.133
g
liver
sample
was
analyzed
for
total
fluorine.
The
fluorine
value
calculated
as
PFOS
was
equivalent
to
11
ppm
in
the
liver.

PFOS
values
in
serum
and
liver
are
approximately
the
same
at
the
0.5
mg/
kg/
day
level
but
are
substantially
higher
in
the
liver
at
the
1.5
mg/
kg/
day
level.
Serum
levels
were
not
determined
at
the
4.5
mg/
kg
level.
That
may
be
because
only
a
single
female
survived
until
scheduled
sacrifice
in
the
4.5
mg/
kg/
day
dose
group.
From
this
study,
there
do
not
appear
to
be
any
differences
in
PFOS
in
either
serum
or
liver
between
the
sexes.

The
values
of
PFOS
in
the
liver
and
serum
should
be
viewed
with
caution.
There
were
a
very
limited
number,
one
male
and
one
female,
of
animals
per
group.
Storage
of
the
liver
samples
at
refrigeration
temperature
for
long
periods
of
time
was
not
ideal.
Although
PFOS
is
stable,
the
liver
itself
obviously
underwent
some
deterioration
as
evidenced
by
the
separation
of
a
liquid
phase.
Sample
size
may
also
have
been
inaccurate
since
the
visual
estimation
of
solid
to
liquid
phase
is
not
a
precise
measurement.

In
order
to
determine
the
dose
range
for
a
six­
month
study,
Thomford
et
al.
(
unaudited
draft,
1998)
conducted
a
4­
week
range­
finding
study
in
cynomolgus
monkeys.
Male
and
female
cynomolgus
monkeys
were
administered
doses
of
0
(
2/
sex/
group),
0.02
(
3/
sex/
group),
or
2.0
mg/
kg/
day
(
1/
sex/
group)
PFOS
in
capsules
placed
directly
into
the
stomach.
The
monkeys
weighed
2.1­
2.4
kg
at
study
initiation.
It
appears
the
monkeys
were
observed
at
least
daily
for
general
clinical
signs
and
body
weights
were
recorded
twice
ENV/
JM/
RD(
2002)
17/
FINAL
31
weekly.
Hematological
and
clinical
chemistry
analyses
were
conducted
on
samples
collected
before
the
beginning
of
the
study
at
day
­
7
(
baseline
values)
and
day
29.
Additional
blood
samples
for
clinical
chemistry
were
collected
on
study
days
2,
7,
and
14.
Blood
samples
for
serum
PFOS
concentrations
were
taken
on
days
7,
2,
3,
7,
14,
and
29.
In
addition,
samples
from
day
­
7
and
day
29
were
analyzed
for
levels
of
estradiol,
estrone,
estriol,
thyroid
stimulating
hormone,
triiodothyronine,
and
thyroxin.
The
study
animals
were
terminated
as
scheduled
at
30
days.
At
necropsy
a
sample
of
liver
was
collected
from
each
animal
for
palmitoyl
CoA
oxidase
activity
analyses.
Samples
of
liver,
testes,
and
pancreas
were
collected
for
proliferation
cell
nuclear
antigen
evaluation.
A
sample
of
liver
was
also
collected
from
each
animal
for
PFOS
concentration
analysis.
The
following
organs
from
control
and
all
treated
groups
were
examined
microscopically:
adrenals,
eye,
kidney,
liver,
lung,
spleen,
pancreas,
femoral
bone
marrow,
testes,
and
thymus.

None
of
the
monkeys
died
before
the
study
was
ended.
There
were
no
test­
related
effects
on
clinical
observations,
body
weight,
food
consumption,
body
temperatures,
hematology,
or
macroscopic
or
microscopic
pathology
findings.
No
test­
related
effects
were
noted
in
the
levels
of
estrone,
estriol,
thyroid
stimulating
hormone,
and
thyroxin.
Cell
proliferation,
as
measured
by
immunohistochemical
detection
of
proliferating
cell
nuclear
antigen,
was
not
increased
in
the
liver,
testes,
or
pancreas
of
monkeys.

The
monkeys
in
the
2.0
mg/
kg/
day
group
both
showed
estradiol
levels
that
were
less
than
their
prestudy
values
and
controls
at
day
29.
Similarly,
the
monkeys
in
the
2.0
mg/
kg/
day
group
both
showed
triiodothyronine
levels
that
were
less
than
their
prestudy
values
and
controls
at
day
29.
However,
pretreatment
the
high­
dose
monkeys
also
had
triiodothyronine
values
lower
than
controls
in
baseline
samples
as
well.
Since
the
numbers
of
tested
animals
are
small
and
baseline
levels
are
variable,
it
is
not
clear
if
these
hormone
level
changes
are
treatment­
related
effects.

In
the
final
study,
Thomford
(
2002)
administered
PFOS
to
cynomolgus
monkeys
by
oral
capsule
at
doses
of
0
(
6
monkeys
per
sex),
0.03
(
4
monkeys
per
sex),
0.15
(
6
monkeys
per
sex),
or
0.75
mg/
kg/
day
(
6
monkeys
per
sex)
for
26
weeks.
Two
animals
from
the
control,
0.15
and
0.75
mg/
kg/
day
groups
were
assigned
to
a
recovery
group
and
were
not
treated
for
at
least
52
weeks
following
the
last
administration
of
PFOS.
Animals
were
observed
twice
daily
for
mortality
and
moribundity
and
were
examined
at
least
once
daily
for
abnormalities
and
signs
of
toxicity;
food
consumption
was
assessed
qualitatively.
Ophthalmic
examinations
were
done
before
initiation
of
treatment
and
during
weeks
26
and
52.
Body
weight
data
were
recorded
weekly
before
the
start
of
treatment,
on
Days
 
1
and
1
and
weekly
thereafter.
Blood
and
urine
samples
were
collected
for
clinical
hematology,
clinical
chemistry,
and
urinalysis
before
the
start
of
treatment
and
at
specified
intervals
during
treatment
and
recovery.
Blood
samples
were
also
taken
for
hormone
determinations.
Samples
of
serum
were
collected
at
various
time
points
during
the
study
and
sent
to
3M
for
analysis
of
PFOS
levels.
The
following
organs
were
weighed
at
scheduled
and
unscheduled
sacrifices;
paired
organs
were
weighed
separately:
adrenal
(
2),
brain,
epididymis
(
2),
kidney
(
2),
liver,
ovary
(
2),
pancreas,
testis
(
2),
and
thyroid
(
2)
with
parathyroid.
The
following
tissues
were
collected
for
histopathology:
adrenals
(
2),
aorta,
brain,
cecum,
cervix,
colon,
duodenum,
epididymis
(
2),
esophagus,
eyes
(
2),
femur
with
bone
marrow,
gallbladder,
heart,
ileum,
jejunum,
kidneys
(
2),
lesions,
liver,
lung,
mammary
gland,
mesenteric
lymph
node,
ovary
(
2),
pancreas,
pituitary,
prostate,
rectum,
salivary
gland
[
mandibular
(
2)],
sciatic
nerve,
seminal
vesicle
(
2),
skeletal
muscle
(
thigh),
skin,
spinal
cord
(
cervical,
thoracic,
and
lumbar),
spleen,
sternum
with
bone
marrow,
stomach,
testis
(
2),
thymus,
thyroid
(
2)
with
parathyroid,
trachea,
urinary
bladder,
uterus,
and
vagina.
Liver
specimens
from
the
0.15
and
0.75
mg/
kg/
day
recovery
animals
were
collected
via
biopsy
and
analyzed
for
PFOS
levels.

Males
weighed
3.3­
3.4
kg
and
females
weighed
2.8­
2.9
kg
at
the
beginning
of
the
study.
At
the
end
of
26
weeks
of
treatment,
males
weighed
3.7,
3.8,
3.5,
and
3.3
kg
for
the
0,
0.03,
0.15
and
0.75
mg/
kg/
day
treatment
groups
respectively.
Females
weighed
3.1,
3.1,
3.1
and
2.8
kg
for
the
0,
0.03,
0.15
and
0.75
mg/
kg/
day
treatment
groups
respectively.
The
difference
between
the
control
and
the
0.75
mg/
kg/
day
ENV/
JM/
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2002)
17/
FINAL
32
female
treatment
groups
was
statistically
significant.
At
the
end
of
the
recovery
period,
differences
in
weight
between
the
control
and
treated
animals
were
no
longer
obvious.

Two
males
from
the
0.75
mg/
kg/
day
group
did
not
survive
to
the
scheduled
sacrifice.
One
animal
died
after
dosing
on
Day
155
(
Week
23).
Clinical
signs
noted
in
this
animal
included:
constricted
pupils,
pale
gums,
few,
mucoid,
liquid
and
black­
colored
feces,
low
food
consumption,
hypoactivity,
labored
respiration,
dehydration,
and
recumbent
position.
In
addition,
the
animal
was
cold
to
the
touch.
An
enlarged
liver
was
detected
by
palpation.
Cause
of
death
was
determined
to
be
pulmonary
necrosis
with
severe
acute
inflammation.
On
day
179,
the
second
male
was
sacrificed
in
a
moribund
condition.
Clinical
signs
noted
included
low
food
consumption,
excessive
salivation,
labored
respiration,
hypoactivity
and
ataxia.
The
cause
of
death
was
not
determined.

Males
and
females
in
the
0.75
mg/
kg/
day
dose­
group
had
lower
total
cholesterol
and
males
and
females
in
the
0.15
and
0.75
mg/
kg/
day
groups
had
lower
high
density
lipoprotein
cholesterol
during
treatment.
The
effect
on
total
cholesterol
worsened
with
time.
By
day
182,
mean
total
cholesterol
for
males
and
females
in
the
high
dose
group
were
68%
and
49%
lower,
respectively,
that
than
levels
in
the
control
animals.
The
effect
on
high
density
lipoprotein
was
greater
than
that
seen
with
cholesterol.
On
day
182,
the
mean
high
density
lipoprotein
levels
were
79%
and
62%
lower,
respectively
in
males
and
females
from
the
high
dose
group
than
they
were
in
male
and
female
control
animals.
Males
in
the
high
dose
group
also
had
lower
total
bilirubin
concentrations
and
higher
serum
bile
acid
concentrations
than
males
in
either
the
control
or
other
treatment
groups.
The
effect
on
total
cholesterol
was
reversed
within
5
weeks
of
recovery
and
the
effect
on
high
density
lipoprotein
cholesterol
was
reversed
within
9
weeks
of
recovery.

Estradiol
values
were
lower
in
males
given
0.75
mg/
kg/
day
on
days
62,
91,
and
182
but
because
of
variation
only
the
day
182
value
was
significant.
Estrone
values
were
generally
higher
in
the
treated
females
on
days
37,
62
and
91
but
again
because
of
variation
in
the
data
none
of
these
values
were
significantly
different
from
the
controls.
Except
for
males
in
the
0.15
mg/
kg/
day
group,
triiodothyronine
values
were
significantly
lower
on
days
91
and
182
in
males
and
females
given
0.15
and
0.75
mg/
kg/
day.
There
were
other
instances
in
which
hormone
values
in
treated
groups
were
different
from
those
of
controls
but
these
differences
were
not
consistent
over
time
or
between
sexes,
were
not
clearly
dose­
related
and
did
not
appear
to
be
related
to
the
administration
of
the
test
material.
Apparent
differences
in
the
sexual
maturity
of
both
males
and
females
used
in
the
study
complicates
the
interpretation
of
the
hormone
data.

At
terminal
sacrifice,
females
in
the
0.75
mg/
kg/
day
dose­
group
had
increased
absolute
liver
weight,
liverto
body
weight
percentages,
and
liver­
to­
brain
weight
ratios.
In
males,
liver­
to
body
weight
percentages
were
increased
in
the
high­
dose
group
compared
to
the
controls.
"
Mottled"
livers
were
observed
in
two
high­
dose
males
and
in
one
high­
dose
female.
Of
the
two
males
not
surviving
until
the
scheduled
terminal
sacrifice,
one
had
a
"
mottled"
and
large
liver.
Three
of
4
high­
dose
males
(
including
those
that
did
not
survive
to
scheduled
sacrifice)
had
centrilobular
or
diffuse
hepatocellular
hypertrophy
that
was
also
observed
in
all
high­
dose
females.
Centrilobular
or
diffuse
hepatocellular
vacuolation
occurred
in
2
of
4
females
and
2
of
4
males
in
the
high­
dose
group.

No
PFOS
related
lesions
were
observed
either
macroscopically
or
microscopically
at
recovery
sacrifice
indicating
that
the
effects
seen
at
terminal
sacrifice
may
be
reversible.

Serum
and
liver
specimens
collected
from
test
animals
were
sent
to
the
3M
Laboratory
and
analyzed
for
the
presence
of
PFOS.
Serum
was
harvested
from
blood
that
was
centrifuged
within
one
hour
of
collection.
Liver
specimens
were
flash
frozen
in
liquid
nitrogen.
Both
liver
and
serum
samples
were
stored
in
a
freezer
set
to
maintain
specimens
at
­
60
to
 
80oC
until
shipped
to
the
3M
Lab.
Samples
were
shipped
frozen
and
on
dry
ice
from
Covance
Laboratories
to
3M
periodically
from
August
1998
through
March
2000
which
covered
the
in­
life
phase
of
the
study.
Once
received
at
3M
specimens
were
stored
in
freezer
at
either
 
ENV/
JM/
RD(
2002)
17/
FINAL
33
55oC
±
10­
20oC
or
 
20oC
±
10oC.

During
the
first
26
weeks
of
the
study
a
total
of
550
serum
specimens
and
30
liver
specimens
were
collected.
Of
the
serum
specimens,
151
were
from
Group
1,
99
from
Group
2,
152
from
Group
3
and
148
from
Group
4.
Eight
liver
samples
were
collected
from
Group
1,
8
from
Group
2,
12
from
Group
3
and
14
from
Group
4.
In
the
recovery
Groups,
72
serum
and
4
liver
samples
were
collected
from
Group
1;
72
serum
and
4
liver
samples
from
Group
3
and
80
serum
and
4
liver
samples
from
Group
4.

Liver
and
serum
samples
were
extracted
using
an
ion­
pairing
reagent
and
methyl­
tert­
butyl
ether
(
MtBE).
Liver
samples
were
homogenized
prior
to
extraction.
Sample
extracts
were
analyzed
using
high­
pressure
liquid
chromatography­
electrospray/
tandem
mass
spectrometry
(
HPLC­
ES/
MS/
MS)
in
the
multiple
response
mode.
PFOS
levels
were
quantitated
by
external
standard
calibration.

Liver
samples
were
homogenized
in
water.
An
aliquot
of
each
liver
homogenate
and
all
serum
samples
were
spiked
with
THPFOS
and
extracted
using
an
ion­
pairing
extraction
procedure.
An
ion­
pairing
reagent
was
added
to
the
samples
and
the
ion
pairs
were
partitioned
into
MtBE.
The
extracts
were
evaporated
until
dry
on
a
nitrogen
evaporator
and
then
were
reconstituted
in
1.0
mL
of
methanol
and
passed
through
a
0.2
µ
m
nylon
filter.

The
analyses
were
performed
by
monitoring
one
or
more
product
ions
selected
from
a
single
primary
ion
characteristic
of
the
fluorochemical
of
interest
using
HPLC/
ES/
MS/
MS.
Molecular
ion
499,
the
primary
ion
for
PFOS
(
C8F17SO3
­)
analysis,
was
fragmented
to
produce
ion
99
(
FSO3
­).
Ion
99
was
monitored
for
quantitative
analysis.

Although
low
levels
of
PFOS
were
often
detected
in
the
sera
and
liver
of
the
control
animals,
these
levels
were
significantly
lower
than
those
found
in
the
low
dose
test
animals.
PFOS
levels
in
the
sera
of
test
animals
increased
with
dose
during
treatment
from
21.0
±
1.57
and
20.4
±
2.71
µ
g/
ml
in
the
Group
4
males
and
females
respectively
at
the
end
of
Week
1
to
194
±
8.94
and
160
±
23.1
µ
g/
ml
in
males
and
females
respectively
in
Group
4
at
the
end
of
Week
27.
During
recovery,
PFOS
levels
in
serum
samples
decreased
over
time
until
they
reached
41.1
±
25.9
µ
g/
ml
in
males
and
41.4
±
1.15
µ
g/
ml
in
females
from
Group
4
at
79
weeks
post­
treatment.
Control
values
were
<
LOQ
(
the
limit
of
quantitation)
at
Week
4
in
both
males
and
females
and
0.0215
±
0.00296
and
0.0243
±
0.00355
µ
g/
ml
in
males
and
females
respectively
at
the
end
of
Week
79.
The
serum
values
for
selected
weeks
of
treatment
and
recovery
are
shown
in
the
table
below.
There
were
no
significant
differences
between
PFOS
levels
in
the
sera
of
treated
males
and
females.
ENV/
JM/
RD(
2002)
17/
FINAL
34
Table
2.
Average
PFOS
Concentrations
(
µ
g/
ml)
in
Serum
of
Monkeys
for
Selected
Weeks
During
Treatment
and
Recovery
Group
1
0.0
mg/
kg/
day
Group
2
0.03
mg/
kg/
day
Group
3
0.15
mg/
kg/
day
Group
4
0.75
mg/
kg/
day
Males
Females
Males
Females
Males
Females
Males
Females
Week
1
<
LOQ
<
LOQ
0.869
±
0.147
0.947
±
0.110
4.60
±
0.782
3.71
±
0.455
21.0
±
1.57
20.4
±
2.71
Week
4
<
LOQ
<
LOQ
3.20
±
0.577
3.40
±
0.291
17.8
±
1.68
16.5
±
1.87
95.3
±
70.4
92.7
±
39.6
Week
16
0.0407
±
0.0110
0.0432
±
0.0081
11.2
±
2.44
10.5
±
1.90
56.2
±
5.84
42.1
±
4.04
189
±
15.9
162
±
19.3
Week
27
0.0529
±
0.0145
0.0416
±
0.0148
15.9
±
5.54
11.1
±
1.52
68.1
±
5.75
58.5
±
4.67
194
±
8.93
160
±
23.9
Week
35
0.0459
±
0.00303
0.0723
±
0.00352
Not
Determined
Not
Determined
84.5
±
12.0
74.7
±
9.53
181
±
19.5
171
±
10.1
Week
47
0.0355
±
0.00221
0.0459
±
0.00323
Not
Determined
Not
Determined
48.3
±
3.69
42.6
±
6.70
124
±
25.9
98.3
±
8.32
Week
57
0.0327
±
000526
0.0445
±
0.00385
Not
Determined
Not
Determined
30.2
±
2.36
32.3
±
1.34
78.0
±
16.3
106
±
3.84
Week
69
0.0406
±
0.00313
0.0400
±
0.00301
Not
Determined
Not
Determined
26.4
±
2.59
34.5
±
3.46
84.0
±
52.4
75.0
±
5.25
Week
79
0.0215
±
0.00296
0.0243
±
0.00355
Not
Determined
Not
Determined
19.1
±
0.805
21.4
±
2.01
41.1
±
25.9
41.4
±
1.15
LOQ
=
Lowest
Observable
Concentration
Liver
values
behaved
in
a
manner
similar
to
serum
values
and
increased
over
time.
At
Week
27
mean
PFOS
values
on
an
RSD
basis
were
22.2
±
0.0269
in
Group
1
males
and
16.8
±
0.0178
in
females
in
Group
1;
27.0
±
4.66
and
9.73
±
2.15
in
males
and
females
in
Group
2;
33.1
±
19.5
in
males
and
21.4
±
14.9
in
females
in
Group
3,
and
6.03
±
23.9
in
males
and
5.00
±
13.6
in
females
in
Group
4.
At
Week
79
values
in
the
liver
were
71.0
±
33.4
in
males
and
21.4
±
10.8
in
females
in
Group
4.
At
Week
80,
values
were
14.9
±
1.38
in
Group
3
males
and
23.5
±
4.98
in
Group
3
females.

3.5
Carcinogenicity
The
chronic
toxicity
and
carcinogenicity
of
perfluorooctane
sulfonic
acid
potassium
salt
(
PFOS;
T­
6295)
have
been
studied
in
rats
(
3M,
2002).
The
results
of
the
study
show
that
PFOS
is
hepatotoxic
and
carcinogenic,
inducing
tumors
of
the
liver,
and
of
the
thyroid
and
mammary
glands.
Based
on
the
liver
toxicity,
the
no­
observed­
adverse­
effect
level
(
NOAEL)
for
PFOS
is
considered
to
be
0.5
ppm
in
male
rats
and
2
ppm
in
female
rats;
the
low
observed­
adverse­
effect
level
(
LOAEL)
is
2
ppm
in
male
rats
and
5
ppm
in
female
rats.

In
this
study,
groups
of
40­
70
male
and
female
Crl:
CD
(
SD)
IGS
BR
rats
were
given
PFOS
in
the
diets
at
concentrations
of
0.5,
2,
5,
or
20
ppm
for
104
weeks.
A
control
group
was
given
diets
containing
acetone,
the
vehicle.
A
recovery
group
was
given
the
test
material
at
20
ppm
for
52
weeks
and
was
observed
till
ENV/
JM/
RD(
2002)
17/
FINAL
35
death.
Five
animals/
sex
in
the
treatment
groups
were
sacrificed
during
weeks
4,
14
and
53;
liver
samples
were
collected
for
mitochondrial
activity,
hepatocellular
proliferation
rate,
and
determination
of
palmitoyl­
CoA
oxidase
activity.
Serum
and
liver
specimens
were
collected
for
analyses
of
the
presence
and
concentration
of
PFOS
in
liver
and
serum
during
and
at
the
end
of
the
in­
life
phase
of
the
study.

There
was
a
significant
increased
trend
in
survival
that
occurred
in
the
males
that
was
due
to
significant
increases
in
survival
in
mid­
high
(
5.0
ppm)
and
high­
dose
(
20.0
ppm)
groups
as
compared
to
that
of
the
control
group.
None
of
the
other
treated
groups
in
the
males
revealed
any
significant
differences
in
survival.
No
significant
trend
was
noted
in
survival
in
females.
There
was
a
significant
decrease
in
survival
in
the
mid­
dose
(
2.0
ppm)
group
and
not
in
the
mid­
high
(
5.0
ppm)
and
high­
dose
(
20.0
ppm)
groups
as
compared
to
that
of
the
control.

Males
given
20
ppm
had
significantly
lower
mean
body
weights
compared
to
animals
in
the
control
group
during
weeks
9
through
37.
Females
given
20
ppm
had
significantly
lower
body
weights
compared
to
animals
in
the
control
group
during
weeks
3
through
101.
At
week
105,
mean
body
weights
of
the
surviving
males
and
females
were
not
significantly
different
from
the
controls.
Food
consumption
for
males
and
females
was
similar
in
all
treated
groups
compared
to
animals
given
the
control
material
except
for
the
high­
dose
females
which
had
statistically
significantly
lower
food
consumption
during
weeks
2
through
44.

At
the
week
14
and
week
53
interim
sacrifice,
absolute
and
relative
liver
weights
were
significantly
increased
in
the
males
given
20
ppm.
In
females
given
20
ppm,
only
the
liver­
to­
body
weight
percentage
was
significantly
increased.
Treatment­
related
histomorphologic
changes
were
seen
in
the
liver
in
the
males
given
5
or
20
ppm
and
in
the
females
given
20
ppm.
The
changes
consisted
of
hypertrophy
of
hepatocytes
in
centrilobular
areas
in
males
and
females,
and
midzonal
to
centrilobular
hepatocytic
vacuolation.
The
incidence
and
severity
of
the
changes
tended
to
be
greater
in
the
males.
Dietary
administration
of
PFOS
for
approximately
53
weeks
was
associated
with
mildly
to
moderately
lower
cholesterol
for
males
and
females
fed
20
ppm;
and
mildly
higher
alanine
aminotransferase
for
males
fed
20
ppm.
In
the
unscheduled
sacrifices
between
Weeks
54
and
105,
animals
given
20
ppm
had
increased
hepatocellular
centrilobular
hypertrophy,
eosinophilic
hepatocytic
granules,
and
centrilobular
hepatocytic
pigment
were
noted.
Increased
hepatocellular
centrilobular
hypertrophy
was
seen
in
animals
given
5
ppm.

At
the
terminal
sacrifice,
the
livers
of
animals
given
5
or
20
ppm
exhibited
a
slight
increase
in
macroscopic
findings,
including
enlarged,
mottled,
diffuse
darkened,
or
focally
lightened.
Hepatotoxicity,
characterized
by
significant
increases
(
P<
0.05)
in
centrilobular
hypertrophy,
centrilobular
eosinophilic
hepatocytic
granules,
centrilobular
hepatocytic
pigment,
or
centrilobular
hepatocytic
vacuolation
was
noted
in
male
and/
or
female
rats
given
5
or
20
ppm.
A
significant
increase
(
P<
0.05)
in
hepatocellular
centrilobular
hypertrophy
was
also
observed
in
mid­
dose
(
2
ppm)
male
rats.
Significant
increases
in
the
incidence
of
cystic
hepatocellular
degeneration
was
found
in
all
the
male
treated
groups
(
0.5,
2,
5,
or
20
ppm);
however,
this
lesion
is
believe
to
be
due
to
old
age
of
the
animals
and
is
not
considered
to
be
treatment­
related.

Based
on
the
pathological
findings
in
the
liver,
the
no­
observed­
adverse­
effect
level
(
NOAEL)
for
PFOS
is
considered
to
be
0.5
ppm
in
male
rats
and
2
ppm
in
female
rats;
the
low
observed­
adverse­
effect
level
(
LOAEL)
is
2
ppm
in
male
rats
and
5
ppm
in
female
rats.

There
was
no
effect
on
hepatic
palmitoyl­
CoA
oxidase
activity.
There
were
also
no
statistically
significant
increases
in
cell
proliferation
as
measured
by
proliferative
cell
nuclear
antigen
(
PCNA)
at
weeks
4
and
14,
or
by
bromodeoxyuridine
(
BrdU)
at
week
53.

For
neoplastic
effects,
a
significant
positive
trend
(
P=
0.0276)
was
noted
in
the
incidences
of
hepatocellular
adenoma
in
male
rats.
This
was
due
to
a
significant
increase
(
P<
0.05)
in
the
high­
dose
group
(
11.7%,
7/
60)
ENV/
JM/
RD(
2002)
17/
FINAL
36
over
the
control
(
0%,
0/
60).
A
significantly
increased
incidence
(
P<
0.05)
was
observed
for
thyroid
follicular
cell
adenoma
in
the
high­
dose
recovery
group
(
23.1%,
9/
39)
when
compared
to
the
control
group
(
5%,
3/
60).
There
was
also
a
slight
increase
in
the
combined
thyroid
follicular
cell
adenoma
and
carcinoma
in
the
high­
dose
recovery
group
(
25.6%,
10/
39)
as
compared
to
that
of
the
control
group
(
10%,
6/
60);
the
increase
did
not
reach
statistical
significance
relative
to
the
control
but
did
reach
statistical
significance
relative
to
the
high­
dose
group
(
8.5%,
5/
59).

In
the
females,
significant
positive
trends
were
observed
in
the
incidences
of
hepatocellular
adenoma
(
P=
0.0153)
and
combined
hepatocellular
adenoma
and
carcinoma
(
P=
0.0057).
These
cases
were
due
to
significant
increases
in
the
high­
dose
group
(
8.3%,
5/
60,
and
10%,
6/
60)
as
compared
to
the
control
(
0%,
0/
60).
A
significant
increase
(
P=
0.0471)
for
combined
thyroid
follicular
cell
adenoma
and
carcinoma
was
observed
in
the
mid­
high
(
5.0
ppm)
group
(
6%,
3/
50)
as
compared
to
the
control
group
(
0%,
0/
60).
Except
for
the
high­
dose
group
(
which
showed
a
slight
decrease
in
incidences
of
mammary
fibroadenoma/
adenoma
and
combined
mammary
fibroadenoma
and
carcinoma),
increases
in
mammary
tumors
were
observed
in
all
treatment
groups
when
compared
to
the
controls.
Significant
increases
(
P<
0.05)
in
mammary
fibroadenoma/
adenoma
(
60%,
30/
50)
and
combined
mammary
fibroadenoma/
adenoma
and
carcinoma
(
72%,
36/
50)
were
observed
in
the
low
 
dose
(
0.5
ppm)
group
as
compared
to
the
respective
controls
(
38.3%,
23/
60
and
48.3%,
29/
60).
The
mid­
dose
(
2.0
ppm)
group
also
exhibited
a
statistically
significant
(
P<
0.05)
increase
(
64.6%,
31/
48)
in
the
incidence
of
combined
mammary
fibroadenoma/
adenoma/
carcinoma
over
the
control
group
(
43.8%,
29/
60).
Increases
in
mammary
tumors
in
the
mid­
high
(
5
ppm)
dose
group
did
not
reach
statistical
significance
relative
to
the
control.
ENV/
JM/
RD(
2002)
17/
FINAL
37
The
carcinogenicity
data
of
PFOS
in
rats
are
summarized
in
Table
3.

Table
3.
Summary
of
carcinogenicity
data
of
PFOS
in
rats.

Tumor
incidence
(%)
Tumors
Male
0
0.5
ppm
2
ppm
5
ppm
20
ppm
20
ppm
recovery#
Liver
Hepatocellular
adenoma*
0
(
0/
60)
6.0
(
3/
50)
6.0
(
3/
50)
2.0
(
1/
50)
11.7**
(
7/
60)
Thyroid
Follicular
cell
adenoma
Follicular
cell
carcinoma
Combined
5.0
(
3/
60)
5.0
(
3/
60)
10.0
(
6/
60)
10.2
(
5/
49)
2.0
(
1/
49)
12.2
(
26/
49)
8.0
(
4/
50)
2.0
(
1/
50)
10.0
(
5/
50)
8.2
(
4/
49)
4.1
(
2/
49)
10.2
(
5/
49)
6.8
(
4/
59)
1.7
(
1/
59)
8.5
(
5/
59)
23.1**
(
9/
39)
2.6
(
1/
39)
25.6
(
10/
39)

Female
Liver
Hepatocellular
adenoma*
Hepatocellular
carcinoma
Combined*
0
(
0/
60)
0
(
0/
60)
0
(
0/
60)
2.0
(
1/
50)
0
(
0/
50)
2.0
(
1/
50)
2.0
(
1/
49)
0
(
0/
49)
2.0
(
1/
49)
2.0
(
1/
50)
0
(
1/
50)
2.0
(
1/
50)
8.3**
(
5/
60)
1.7
(
1/
60)
10.0**
(
6/
60)
Thyroid
Follicular
cell
adenoma
Follicular
cell
carcinoma
Combined
0
(
0/
60)
0
(
0/
60)
0
(
0/
60)
0
(
0/
50)
0
(
0/
50)
0
(
0/
50)
0
(
0/
49)
0
(
0/
49)
0
(
0/
49)
4.0
(
2/
50)
2.0
(
1/
50)
6.0**
(
3/
50)
1.7
(
1/
60)
0
(
0/
60)
1.7
(
1/
60)
Mammary
Fibroadenoma/
adenoma
Carcinoma
Combined
38.3
(
23/
60)
18.3
(
11/
60)
48.3
(
29/
60)
60.0**
(
30/
50)
24.0
(
12/
50)
72.0**
(
36/
50)
45.8
(
22/
48)
31.2
(
15/
48)
64.6**
(
31/
48)
52.04
(
26/
50)
22.0
(
11/
50)
58,0
(
29/
50)
25
(
15/
60)
23.3
(
14/
60)
40.0
(
24/
60)

*
Significant
positive
trend
(
P
<
0.03).
**
Significantly
increased
over
the
control
(
P
<
0.05).
#
Recovery
group;
after
52
weeks
of
treatment.

Serum
and
Liver
level
of
PFOS
Under
the
conditions
of
the
studies,
PFOS
was
observed
in
the
serum
and
liver
of
rats
dosed
with
perfluorooctane
sulfonic
acid
potassium
salt
(
PFOS
T­
6295).
Trace
levels
of
PFOS
were
often
detected
in
the
serum
and
liver
of
the
control
animals.
Detailed
specimen
data
are
presented
in
Table
4
and
Table
5.
ENV/
JM/
RD(
2002)
17/
FINAL
38
Table
4.
Summary
of
PFOS
Concentration­
Serum
(
ug/
mL)

a
Not
corrected
for
purity
of
the
standard
material.
b
LOQ­
Limit
of
Quanfitation
=
0.00910
pg/
mL
c
LOQ­
Limit
of
Quanfitation
=
0.0457
pg/
mL
d
C92987F
sample
spilled
during
extraction,
no
sample
remaining
for
analysis.
It
is
not
possible
to
verify
true
recovery
of
endogenous
analyte
from
tissues
without
radio­
labeled
reference
material.
The
only
measurement
of
accuracy
available
at
this
time,
matrix
spike
studies,
indicated
(
that
the
sera
data
are
accurate
to
±
30%;
liver
data
are
accurate
to
±
50%.
Group
6
Timepoint
Sex
0
ppm
0.5
ppm
2
ppm
5
ppm
20
ppm
High
Recovery
Average
±
SD
Average
±
SD
Average
±
SD
Average
±
SD
Average
±
SD
Average
±
SD
Male
<
LOQb
(
n=
5)
0.907
±
0.0619
(
n=
5)
4.33
±
1.16
(
n=
5)
7.57
±
2.17
(
n=
5)
41.8
±
7.92
(
n=
5)
Week
0
Female
0.0259
±
0.00663
(
n
=
5)
1.61
±
0.207
(
n
=
5)
6.62
±
0.499
(
n
=
5)
12.6
±
1.73
(
n
=
5)
54.0
±
7.34
(
n
=
5)

Male
<
LOQc
(
n
=
5)
4.04
±
0.801
(
n
=
5)
17.1
±
1.22
(
n
=
5)
43.9
±
4.90
(
n
=
5)
148
±
13.8
(
n
=
5)
Week
14a
Female
2.67
±
4.58
(
n
=
5)
6.96
±
0.993
(
n
=
4d)
27.3
±
2.34
(
n
=
5)
64.4
±
5.48
(
n
=
5)
223
±
22.4
(
n
=
5)

Male
0.0249
±
0.0182
(
n
=
5)
146
±
33.5
(
n
=
4)
Week
53
Female
0.395
±
0.777
(
n
=
5)
220
±
44.0
(
n
=
5)

Male
Day
719
Female
20.2
±
13.3
(
n
=
9)

Male
0.0118
±
0.0104
(
n
=
11)
1.31
±
1.30
(
n
=
10)
7.60
±
8.60
(
n
=
17)
22.5
±
23.5
(
n
=
25)
69.3
±
57.9
(
n
=
22)
Week
105
Female
0.0836
±
0.134
(
n
=
24)
4.35
±
2.78
(
n
=
15)
75.0
±
45.7
(
n
=
15)
233
±
124
(
n
=
25)

Male
2.42
±
5.09
(
n
=
10)
Week
106
Female
9.51
±
8.70
(
n
=
17)
ENV/
JM/
RD(
2002)
17/
FINAL
39
Table
5.
Summary
of
PFOS
Concentration­
Liver
(
ug/
g)

Group
1
Group
2
Group
3
Group
4
Group
5
Group
6
Timepoint
Sex
Control
Low
Mid
Mid­
High
High
High
Recovery
Average
±
SD
Average
±
SD
Average
±
SD
Average
±
SD
Average
±
SD
Average
±
SD
Male
0.104
±
0.0673
(
n
=
5)
11.0
±
2.31
(
n
=
5)
31.3
±
5.84
(
n
=
5)
47.6
±
12.5
(
n
=
5)
282
±
45.3
(
n
=
5)
Week
0
Female
0.107
±
0.0486
(
n
=
5)
8.71
±
0.552
(
n
=
5)
25.0
±
6.11
(
n
=
5)
83.0
±
14.1
(
n
=
5)
373
±
44.1
(
n
=
5)

Male
0.459
±
0.0573
(
n
=
5)
23.8
±
3.45
(
n
=
5)
74.0
±
6.16
(
n
=
5)
358
±
28.8
(
n
=
5)
568
±
107
(
n
=
5)
Week
10
Female
12.0
±
22.4
(
n
=
5)
19.2
±
3.77
(
n
=
5)
69.2
±
3.46
(
n
=
5)
370
±
22.3
(
n
=
5)
635
±
49.0
(
n
=
5)

Male
0.635
±
1.04
(
n
=
10)
435
±
96.9
(
n
=
9)
Week
53
Female
0.923
±
1.77
(
n
=
10)
560
±
180
(
n
=
10)

Male
Day
719
Female
55.1
±
31.5
(
n
=
9)

Male
0.114
±
0.148
(
n
=
11)
7.83
±
7.34
(
n
=
10)
26.4
±
20.4
(
n
=
17)
70.5
±
63.1
(
n
=
25)
189
±
141
(
n
=
22)
Week
105
Female
0.185
±
0.184
(
n
=
24)
12.9
±
6.81
(
n
=
15)
131
±
61.4
(
n
=
15)
381
±
176
(
n
=
25)

Male
3.12
±
5.97
(
n
=
10)
Week
106
Female
12.9
±
10.4
(
n
=
17)

It
is
not
possible
to
verify
true
recovery
of
endogenous
analyte
from
tissues
without
radio­
labeled
reference
material.
The
only
measurement
of
accuracy
available
at
this
time,
matrix
spike
studies,
indicated
that
the
sera
data
are
accurate
to
±
30%;
liver
data
are
accurate
to
±
50%.

3.6
Developmental
Toxicity
Three
prenatal
developmental
toxicity
studies
of
PFOS
have
been
conducted,
two
studies
in
rats
and
one
study
in
rabbits.
In
addition,
preliminary
results
are
available
for
developmental
toxicity
studies
in
rats
and
mice.

The
first
study
administered
four
groups
of
22
time­
mated
Sprague­
Dawley
rats
0,
1,
5,
and
10
mg/
kg/
day
PFOS
in
corn
oil
by
gavage
on
gestation
days
(
GD)
6­
15
(
Gortner,
1980).
Doses
were
adjusted
according
to
body
weight.
Dams
were
monitored
on
GD
3­
20
for
clinical
signs
of
toxicity.
Individual
body
weights
were
recorded
on
GD
3,
6,
9,
12,
15,
and
20.
Animals
were
sacrificed
on
GD
20
by
cervical
dislocation
and
the
ovaries,
uteri
and
contents
were
examined
for
the
number
of
corpora
lutea,
number
of
viable
and
non­
viable
fetuses,
number
of
resorption
sites,
and
number
of
implantation
sites.
Fetuses
were
weighed
and
sexed
and
subjected
to
external
gross
necropsy.
Approximately
one­
third
of
the
fetuses
were
fixed
in
Bouin's
solution
and
examined
for
visceral
abnormalities
by
free­
hand
sectioning.
The
remaining
fetuses
were
subjected
to
a
skeletal
examination
using
alizarin
red.
ENV/
JM/
RD(
2002)
17/
FINAL
40
Signs
of
maternal
toxicity
consisted
of
significant
reductions
in
mean
body
weights
during
GD
12­
20
at
the
high­
dose
group
of
10
mg/
kg/
day.
No
other
signs
of
maternal
toxicity
were
reported.
Under
the
conditions
of
the
study,
a
NOAEL
of
5
mg/
kg/
day
and
a
LOAEL
of
10
mg/
kg/
day
for
maternal
toxicity
were
indicated.

Developmental
toxicity
evident
at
10
mg/
kg/
day
consisted
of
reductions
in
the
mean
number
of
implantation
sites,
corpora
lutea,
resorption
sites
and
the
mean
numbers
of
viable
male,
female,
and
total
fetuses,
but
the
differences
were
not
statistically
significant.
In
addition,
unusually
high
incidences
of
unossified,
assymetrical,
bipartite,
and
missing
sternebrae
were
observed
in
all
dose
groups;
however,
these
skeletal
variations
were
also
observed
in
control
fetuses
at
the
same
rate
and
therefore
these
effects
were
not
considered
to
be
treatment­
related.
The
most
notable
sign
of
developmental
toxicity
observed
in
all
dose
groups
consisted
of
abnormalities
of
the
lens
of
the
eye,
which
was
not
seen
in
controls.
The
proportion
of
fetuses
with
the
lens
abnormality
in
one
or
both
lenses
was
significantly
higher
in
the
high
dose
group.
All
eye
abnormalities
appeared
to
be
localized
to
the
area
of
the
embryonal
lens
nucleus,
although
a
variety
of
morphological
appearances
were
present
within
that
location.
According
to
the
authors,
this
abnormality
appeared
to
be
an
arrest
in
development
of
the
primary
lens
fibers
forming
the
embryonal
lens
nucleus.
Secondary
lens
fiber
development
progressed
normally
except
immediately
surrounding
the
abnormal
embryonal
nucleus.
Under
the
conditions
of
the
study,
a
LOAEL
for
developmental
toxicity
of
1
mg/
kg/
day
was
indicated;
a
developmental
NOAEL
could
not
be
established.

In
a
second
prenatal
developmental
toxicity
study,
groups
of
25
pregnant
Sprague­
Dawley
rats
were
administered
0,
1,
5,
and
10
mg/
kg/
day
PFOS
in
corn
oil
by
gavage
on
gestation
days
(
GD)
6­
15
(
Wetzel,
1983).
Sexually
mature
Sprague­
Dawley
rats,
one
per
sex
per
cage,
were
paired
until
confirmation
of
mating
or
until
two
weeks
had
elapsed.
Mating
was
confirmed
by
daily
vaginal
examinations
for
the
presence
and
viability
of
sperm
or
the
presence
of
a
copulatory
plug.
The
day
of
confirmation
of
mating
was
designated
as
day
0
of
gestation.
Doses
were
adjusted
according
to
the
most
recently
recorded
body
weight
measurements.
Dams
were
observed
twice
daily
for
signs
of
mortality
and
moribundity
and
once
daily
for
clinical
signs
of
toxicity.
Individual
body
weights
and
food
consumption
were
recorded
on
GD
6,
8,
12,
16,
and
20.
Animals
were
sacrificed
on
GD
20
by
CO2
asphyxiation
and
the
fetuses
were
delivered
by
cesarean
section
on
GD
20.
A
gross
necropsy
was
performed
on
all
dams.
The
uterus
from
each
female
was
excised,
weighed
and
examined
for
the
number
and
placement
of
implantation
sites,
number
and
of
live
and
dead
fetuses,
number
of
early
and
late
resorptions,
and
any
abnormalities
and
then
weighed
again
after
the
contents
were
removed.
The
ovaries
were
examined
for
the
number
of
corpora
lutea.
Each
female
was
examined
by
gross
necropsy.
Each
fetus
was
sexed,
weighed,
and
examined
externally.
Approximately
one­
third
of
the
fetuses
were
fixed
in
Bouin's
solution
and
examined
for
visceral
abnormalities
by
the
Wilson
technique,
with
particular
attention
to
the
eyes,
palate,
and
brain.
The
remaining
fetuses
were
subjected
to
a
skeletal
examination
that
included
evaluation
of
the
skull,
long
bones,
vertebral
column,
rib
cage,
extremities,
and
pectoral
and
pelvic
girdles
using
alizarin
red;
bone
alignment
and
degree
of
ossification
were
also
evaluated.

Evidence
of
maternal
toxicity,
that
was
observed
at
the
5
and
10
mg/
kg/
day
dose
groups
both
during
and
following
treatment
and
considered
to
be
treatment­
related,
consisted
of
hunched
posture,
anorexia,
bloody
vaginal
discharge,
uterine
stains,
alopecia,
rough
haircoat,
and
bloody
crust.
Significant
decreases
in
mean
body
weight
gains
during
GD
6­
8,
6­
16,
and
0­
20
were
also
observed
at
the
5
and
10
mg/
kg/
day
dose
groups.
These
reductions
were
considered
to
be
treatment­
related
since
mean
body
weight
gains
were
greater
than
controls
during
the
post­
exposure
period
(
GD
16­
20).
Significant
decreases
in
mean
total
food
consumption
were
observed
on
GD
17­
20
in
the10
mg/
kg/
day
dose
group,
and
on
GD
7­
16
and
0­
20
in
both
the
5
and
10
mg/
kg/
day
dose
groups.
The
mean
gravid
uterine
weight
in
the
10
mg/
kg/
day
dose
group
was
significantly
lower
when
compared
with
controls.
The
mean
terminal
body
weights
minus
the
gravid
uterine
weights
were
lower
in
all
treated
groups,
with
significant
decreases
at
5
and
10
mg/
kg/
day.
High­
ENV/
JM/
RD(
2002)
17/
FINAL
41
dose
animals
also
exhibited
an
increased
incidence
in
gastrointestinal
lesions.
No
significant
differences
were
observed
in
pregnancy
rates,
number
of
corpora
lutea,
and
number
and
placement
of
implantation
sites
among
treated
and
control
groups.
Two
dams
in
the
10
mg/
kg/
day
dose
group
were
found
dead
on
GD
17.
Under
the
conditions
of
the
study,
a
NOAEL
of
1
mg/
kg/
day
and
a
LOAEL
of
5
mg/
kg/
day
for
maternal
toxicity
were
indicated.

Signs
of
developmental
toxicity
included
a
dose­
related
trend
toward
an
increased
incidence
of
late
resorptions,
total
resorptions,
number
of
dead
fetuses,
and
fetal
loss,
although,
none
of
these
effects
were
statistically
significantly
different
from
controls.
Significant
decreases
in
mean
fetal
weights
for
both
males
and
females
were
observed
in
the
5
and
10
mg/
kg/
day
dose
groups.
The
percent
of
male
fetuses
was
52%,
54%,
and
60%
for
1,
5,
and
10
mg/
kg/
day,
respectively,
compared
to
44%
in
controls.
Statistically
significant
increases
in
incomplete
closure
of
the
skull
were
observed
in
the
low­
and
high­
dose
groups
but
not
in
the
mid­
dose
group.
Statistically
significant
increases
in
the
incidences
in
the
number
of
litters
containing
fetuses
with
visceral
anomalies,
delayed
ossification,
and
skeletal
variations
were
observed
in
the
high
dose
group
of
10
mg/
kg/
day.
These
included
external
and
visceral
anomalies
of
the
cleft
palate,
subcutaneous
edema,
and
cryptorchism
as
well
as
delays
in
skeletal
ossification
of
the
skull,
pectoral
girdle,
rib
cage,
vertebral
column,
pelvic
girdle,
and
limbs.
Skeletal
variations
in
the
ribs
and
sternebrae
were
also
observed.
Under
the
conditions
of
the
study,
a
NOAEL
of
1
mg/
kg/
day
and
a
LOAEL
of
5
mg/
kg/
day
for
developmental
toxicity
were
indicated.

The
developmental
eye
abnormalities
that
were
seen
in
the
previous
study
(
Gortner,
1980)
were
not
observed
in
the
1983
developmental
toxicity
study
even
though
the
study
design
and
doses
were
the
same.
Findings
of
abnormalities
in
eye
development
were
initially
thought
to
be
treatment­
related
but
later
determined
to
be
artifacts
of
sectioning
(
3M
Company,
1999a).

Lau
et
al.
(
2001)
administered
Sprague­
Dawley
rats
and
CD­
1
mice
doses
of
0,
1,
5
or
10
mg/
kg/
day
PFOS
in
0.5%
Tween­
20
by
gavage
beginning
on
gestation
day
2
and
continuing
until
term.
Half
of
the
dams
were
sacrificed
on
gestation
day
21
(
rats)
or
gestation
day
17
(
mice)
and
the
remaining
dams
were
allowed
to
deliver.
Preliminary
results
are
available.
In
rats,
there
was
a
significant
reduction
in
maternal
body
weight
gain
at
5
and
10
mg/
kg/
day.
Maternal
serum
cholesterol
and
triglycerides
were
reduced
at
10
mg/
kg/
day,
but
liver
weights
were
comparable
to
control.
At
10
mg/
kg/
day,
there
was
a
reduction
in
fetal
body
weight
and
an
increase
in
cleft
palate
and
anasarca.
All
pups
were
born
alive,
but
within
4­
6
hours
after
birth
all
the
pups
in
the
10
mg/
kg/
day
group
died,
and
95%
of
the
pups
in
the
5
mg/
kg/
day
group
died
within
24
hours.
In
mice,
maternal
body
weight
was
unaffected
and
liver
weights
were
significantly
increased
at
5
and
10
mg/
kg/
day;
serum
triglycerides
were
reduced
were
elevated
at
5
and
10
mg/
kg/
day.
The
incidence
of
fetal
mortality
was
slightly
increased
at
10
mg/
kg/
day
and
mean
fetal
body
weights
were
comparable
to
control.
However,
neonatal
body
weights
were
reduced
during
the
first
3
days
of
life.
Additional
studies
are
underway
to
further
elucidate
the
dose­
response
relationships
and
to
examine
the
mechanism
for
the
neonatal
death.

Christian
et
al.
(
1999a)
administered
pregnant
New
Zealand
White
rabbits,
22
per
group,
doses
of
0,
0.1,
1.0,
2.5
or
3.75
mg/
kg/
day
PFOS
in
0.5%
Tween­
80
by
gavage
on
gestation
days
7­
20.
A
dose
volume
of
5
mL/
kg
was
administered,
adjusted
daily
on
the
basis
of
individual
body
weights.
The
does
were
observed
twice
daily
for
viability,
and
clinical
observations
were
recorded
1
hour
prior
to
and
after
dosing
during
the
treatment
period
and
once
daily
during
the
post­
treatment
period
(
i.
e.
gestation
days
20­
29).
Maternal
body
weights
were
recorded
on
gestation
days
0
and
6­
29;
food
consumption
was
recorded
daily
throughout
the
study.
On
gestation
day
29,
the
does
were
euthanized;
a
gross
necropsy
of
the
thoracic,
abdominal
and
pelvic
viscera
was
conducted
and
the
number
of
corpora
lutea
in
each
ovary
was
recorded.
The
uteri
were
examined
for
number
and
distribution
of
implantations,
live
and
dead
fetuses,
and
early
and
late
resorptions.
The
fetuses
were
weighed,
sexed
and
examined
for
external
abnormalities.
All
fetuses
were
examined
for
visceral
and
skeletal
abnormalities
and
the
brain
of
one­
half
of
the
fetuses
were
free­
ENV/
JM/
RD(
2002)
17/
FINAL
42
hand
cross­
sectioned
and
examined
in
situ.

In
addition,
a
satellite
study
was
conducted
in
which
pregnant
New
Zealand
White
rabbits
were
administered
the
same
doses
as
in
the
main
study.
The
number
of
does
was
3,
5,
3,
3
and
5
in
the
control,
0.1,
1.0,
2.5
and
3.75
mg/
kg/
day
groups,
respectively.
The
does
were
euthanized
on
gestation
day
21,
blood
samples
were
collected,
and
the
liver
was
weighed
and
sectioned.
The
fetuses
were
removed
and
examined
for
external
abnormalities.
Fetuses
and
placentae
were
pooled
per
litter.
All
samples
were
sent
to
the
Sponsor
(
3M)
for
analysis.
At
this
time,
only
the
liver
and
serum
analyses
have
been
reported
(
3M
Environmental
Laboratory,
2001d).

Maternal
toxicity
was
evident
at
doses
of
1.0
mg/
kg/
day
and
above.
One
doe
in
the
2.5
mg/
kg/
day
group
and
nine
does
in
the
3.75
mg/
kg/
day
aborted.
All
abortions
occurred
on
gestation
days
22­
28
and
were
considered
treatment­
related
by
the
study
authors.
There
was
a
significant
increase
in
the
incidence
of
scant
feces
in
the
3.75
mg/
kg/
day
group.
Scant
feces
were
also
noted
in
one
and
three
does
in
the
1.0
and
2.5
mg/
kg/
day
groups,
respectively.
Mean
maternal
body
weight
gains
were
significantly
reduced
in
the
3.75
mg/
kg/
day
group
on
gestation
days
10­
13,
13­
16,
16­
19
and
21­
24.
Mean
body
weight
gains
were
also
calculated
for
the
treatment
period
(
days
7­
21),
post­
treatment
period
(
days
21­
29)
and
duration
of
the
study
(
days
7­
29).
There
was
a
significant
reduction
in
mean
maternal
body
weight
gain
during
the
treatment
period
in
the
1.0,
2.5
and
3.75
mg/
kg/
day
groups.
Mean
body
weight
gain
for
the
entire
study
period
was
also
significantly
reduced
in
the
2.5
mg/
kg/
day
group.
Mean
food
consumption
(
g/
kg/
day)
was
significantly
reduced
in
the
2.5
mg/
kg/
day
group
on
gestation
days
16­
19,
19­
21
and
21­
24,
as
well
as
for
the
entire
study
period
(
days
7­
29).
Mean
food
consumption
was
significantly
reduced
in
the
3.75
mg/
kg/
day
group
on
gestation
days
13­
16,
16­
19,
19­
21
and
21­
24,
as
well
as
the
entire
treatment
period.
The
LOAEL
for
maternal
toxicity
was
1.0
mg/
kg/
day
and
the
NOAEL
was
0.1
mg/
kg/
day.

Developmental
toxicity
was
evident
at
doses
of
2.5
mg/
kg/
day
and
above.
The
number
of
corpora
lutea,
resorptions,
live/
dead
fetuses,
litter
size
and
sex
ratio
were
comparable
among
treated
and
control
groups.
Mean
fetal
body
weight
(
male,
female
and
sexes
combined)
was
significantly
reduced
in
the
2.5
and
3.75
mg/
kg/
day
groups.
There
was
also
a
significant
reduction
in
the
ossification
of
the
sternum
(
litter
averages)
in
the
2.5
and
3.75
mg/
kg/
day
groups,
and
a
significant
reduction
in
the
ossification
of
the
hyoid
(
litter
averages),
metacarpals
(
litter
averages)
and
pubis
(
litter
and
fetal
averages)
in
the
3.75
mg/
kg/
day
group.
The
LOAEL
for
developmental
toxicity
was
2.5
mg/
kg/
day
and
the
NOAEL
was
1.0
mg/
kg/
day.

In
the
satellite
study
of
does
euthanized
on
gestation
day
21,
the
liver
and
serum
analyses
were
reported
by
3M
Environmental
Laboratory
(
2001d).
All
serum
and
liver
samples
(
including
those
from
untreated
controls)
had
detectable
levels
of
PFOS;
the
values
are
presented
below:

Average
Concentration
of
PFOS
in
Rabbit
Liver
and
Serum
by
Dose
Group
Dose
group
PFOS
conc.
PFOS
conc.
(
mg/
kg/
day)
liver
(
ug/
g)
serum
(
ug/
ml)

0.0
0.239
0.0690
0.1
13.1
2.73
1.0
133
23.8
2.5
317
45.8
3.75
416
88.9
Qualitatively,
increasing
concentrations
of
PFOS
were
found
in
samples
of
liver
and
serum
as
doses
of
PFOS
increased.
The
levels
of
PFOS
are
much
higher
in
the
liver
than
in
the
serum.
ENV/
JM/
RD(
2002)
17/
FINAL
43
These
values
should
be
viewed
with
caution.
It
was
stated
that
because
radio­
labeled
reference
material
was
not
available,
"
it
is
not
possible
to
verify
true
recovery
of
endogenous
analyte
from
tissues."
Matrix
spike
recovery
indicates
the
accuracy
of
quantitation
to
be
+
30%.
It
is
also
noted
that
liver
concentrations
may
be
biased
high.
The
only
conclusion
presented
in
the
laboratory
report
is
that
"
PFOS
was
observed
in
the
liver
and
serum
of
all
rabbits
dosed
with
the
test
article"
(
3M
Environmental
Laboratory,
2001d).

3.7
Reproductive
Toxicity
A
two­
generation
reproductive
toxicity
study,
designed
to
test
for
the
toxic
effects
of
PFOS
on
reproductive
function
in
adult
animals
and
on
developmental,
learning,
and
reproductive
effects
in
the
offspring,
was
conducted
in
Sprague­
Dawley
rats
(
Christian
et
al.,
1999b).
Five
groups
of
35
rats
per
sex
per
dose
group
were
administered
PFOS
by
gavage
at
doses
of
0,
0.1,
0.4,
1.6,
and
3.2
mg/
kg/
day
for
six
weeks
prior
to
and
during
mating.
Treatment
in
male
rats
continued
until
one
day
before
sacrifice
(
approximately
22
days
total);
female
rats
were
treated
throughout
gestation,
parturition,
and
lactation.

F0
Generation:

Parental
animals
(
F0)
were
observed
twice
daily
for
clinical
signs.
Body
weights
and
food
consumption
values
were
recorded
weekly
during
the
treatment
period
in
male
rats;
and
weekly
during
mating
and
then
daily
during
gestation,
and
on
lactation
days
1,
4,
7,
10,
14,
and
at
sacrifice
in
female
rats.
Each
dosage
group
consisted
of
two
sets
of
female
rats.
One
set
consisted
of
the
first
ten
female
rats
with
confirmation
of
mating;
this
group
was
dosed
until
gestation
day
(
GD)
10
and
delivered
via
Caesarean­
sectioning.
The
remaining
females
comprised
the
second
set
which
delivered
naturally.
During
the
21­
day
lactation
period,
the
dams
were
evaluated
for
clinical
signs
during
parturition
and
length
of
gestation,
and
then
each
litter
was
evaluated
at
least
twice
daily
for
size
and
pup
viability
at
birth.
Pup
observations
during
the
21­
day
lactation
period
included
physical
signs,
body
weights,
nursing
behavior,
surface
righting
reflex,
pinna
unfolding,
eye
opening,
acoustic
startle
response
and
air
righting
reflex.
Pupil
constriction
was
evaluated
only
on
lactation
day
21.
On
lactation
day
4,
litters
were
randomly
culled
to
four
male
and
four
female
pups.
The
remaining
pups
were
sacrificed
and
necropsied.
The
F0
male
rats
were
sacrificed
and
necropsied
after
the
end
of
dosing
at
the
time
of
parturition
(
lactation
day
1).
The
testes,
epididymides,
prostate,
and
seminal
vesicles
were
weighed.
Evaluations
of
sperm
number,
motility,
and
morphology
were
not
included
in
the
protocol.
The
F0
generation
females
that
delivered
by
Caesarean­
section
were
sacrificed
on
GD
10
and
necropsied.
Pregnancy
status
was
confirmed,
the
ovaries
were
examined
for
the
number
and
distribution
of
corpora
lutea,
implantation
sites
were
determined,
and
embryos
were
examined
for
viability.
The
F0
generation
females
that
delivered
naturally
were
sacrificed
on
lactation
day
(
LD)
21
and
necropsied.
Ovaries
were
examined
as
above
and
the
number
and
distribution
of
implantation
sites
was
recorded.
The
liver
from
each
parental
rat
was
removed,
weighed
and
analyzed.
Blood
samples
were
collected
from
5
male
rats
that
had
mated
and
from
5
female
rats
on
LD
21
for
pharmacokinetic
analysis;
livers
from
the
pups
from
the
litters
of
these
five
dams
were
also
collected
for
analysis.

F1
Generation:

Since
F1
generation
pup
viability
was
significantly
reduced
in
the
1.6
and
3.2
mg/
kg/
day
dose
groups,
only
the
0.1
and
0.4
mg/
kg/
day
dose
groups
were
carried
into
the
second
generation.
Twenty­
five
F1
generation
rats
per
sex
per
dose
group
were
administered
PFOS
by
gavage
at
doses
of
0,
0.1,
and
0.4
mg/
kg/
day
beginning
on
LD
22
and
continuing
through
the
day
before
sacrifice.
At
24
days
of
age,
one
rat
per
sex
per
litter
in
each
dose
group
was
tested
in
a
passive
avoidance
paradigm.
On
LD
28,
females
were
evaluated
for
the
age
of
vaginal
patency
and
on
LD
34,
male
rats
were
evaluated
for
the
age
of
preputial
separation.
One
rat
per
sex
per
litter
were
evaluated
in
a
water­
filled
M­
maze
on
LD
70.
Assignment
to
cohabitation
within
each
dose
group
began
on
LD
90.
Females
with
evidence
of
mating
were
considered
to
be
at
GD
0
and
assigned
to
individual
housing
for
the
remainder
of
the
dosing
period.
The
F1
generation
male
rats
ENV/
JM/
RD(
2002)
17/
FINAL
44
were
sacrificed
after
mating,
necropsied
and
evaluated
as
described
in
the
F0
generation.
All
F1
generation
females
were
allowed
to
deliver
naturally.
Dams
that
delivered
litters
were
sacrificed
and
necropsied
on
LD
21.
All
F2
generation
pups
were
sacrificed,
necropsied,
and
examined
on
LD
21
as
previously
described
for
the
F1
generation
pups.

In
the
F0
generation
male
rats,
there
were
no
treatment­
related
clinical
signs
of
toxicity,
no
mortality,
and
no
effects
on
mating
or
on
any
of
the
fertility
parameters
evaluated
in
any
dose
group
tested.
Reported
effects
included
reductions
in
both
body
weight
gains
and
in
absolute
and
relative
food
consumption
at
the
1.6
and
3.2
mg/
kg/
day
dose
groups
during
the
pre­
mating
period.
Following
mating,
food
consumption
was
significantly
reduced
in
the
0.4.
and
1.6
mg/
kg/
day
dose
groups.
Terminal
body
weights
were
also
significantly
reduced
in
the
1.6
and
3.2
mg/
kg/
day
dose
groups.
Significant
reductions
in
the
absolute
weights
of
the
seminal
vesicles
(
with
fluid)
and
the
prostate
were
observed
in
F0
males
at
3.2
mg/
kg/
day;
no
other
organ
weight
changes
were
reported.
A
significant
increase
in
the
number
of
males
with
brown
liver
at
3.2
mg/
kg/
day
dose
group
was
also
reported.

In
the
F0
generation
female
rats,
no
deaths
were
reported
at
any
dose
level.
In
dams
sacrificed
on
GD
10
for
Caesarean­
sectioning,
there
did
not
appear
to
be
any
effects
on
estrous
cycling,
mating
and
fertility
parameters,
the
numbers
of
corpora
lutea
and
implantations,
or
in
the
number
of
viable
or
non­
viable
embryos.
The
only
findings
reported
in
the
F0
dams
occurred
in
the
0.4,
1.6,
and
3.2
mg/
kg/
day
dose
groups
and
included
localized
alopecia
during
pre­
mating,
gestation,
and
lactation;
and
reductions
in
body
weight
and
body
weight
gain
and
food
consumption
values
observed
during
the
pre­
mating
period
and
continuing
throughout
gestation
and
lactation.

Reversible
delays
in
reflex
and
physical
development
were
observed
in
the
F1
generation
offspring.
The
ability
to
surface
right
was
significantly
delayed
in
the
1.6
and
3.2
mg/
kg/
day
dose
groups
on
LDs
3­
10
(
delays
in
the
3.2
mg/
kg/
day
dose
group
were
observed
on
LD
1,
after
which
there
were
no
surviving
pups
remaining
for
further
observation).
By
the
end
of
the
observation
period,
however,
all
surviving
pups
in
the
1.6
mg/
kg/
day
dose
group
had
the
ability
to
surface
right.
There
were
no
delays
observed
in
the
ability
to
surface
right
in
dose
groups
<
0.4
mg/
kg/
day.
Similar
responses
were
seen
for
pinna
unfolding
and
eye
opening.
Although
there
were
transient
delays
seen
with
these
signs
of
physical
development
across
all
dose
groups,
by
the
end
of
the
observation
period
responses
in
pups
were
similar
to
controls.
The
time
of
development
of
the
acoustic
startle
reflex
and
the
ability
to
air
right
were
both
significantly
reduced
in
the
1.6
mg/
kg/
day
dose
group.
No
effects
on
these
reflexes
were
observed
in
the
low
dose
group
of
0.1
mg/
kg/
day
and
only
a
transient
delay
(
on
LD
16
only)
in
the
ability
to
air
right
was
seen
in
the
0.4
mg/
kg/
day
group.
At
the
end
of
lactation
(
LD
21),
all
live
pups
in
all
dose
groups
(
0,
0.1,
0.4,
and
1.6
mg/
kg/
day)
had
pupil
constriction
response.

The
most
significant
finding
reported
in
the
offspring
was
that
of
reduced
pup
viability
at
the
two
highest
dose
groups.
The
reductions
in
pup
viability
began
to
appear
on
LD
4
postculling
in
the
1.6
mg/
kg/
day
dose
group,
with
over
26%
of
the
pups
found
dead
between
LD
2­
4.
In
the
3.2
mg/
kg/
day
dose
group
45%
of
the
pups
were
found
dead
on
LD1;
no
pups
survived
beyond
LD
1.
Statistically
significant
increases
in
the
number
of
dams
with
stillborn
pups
were
also
observed
at
3.2
mg/
kg/
day.
As
a
result,
the
viability
index
was
greatly
reduced
in
these
dose
groups
(
0%
at
3.2
mg/
kg/
day
and
66%
at
1.6
mg/
kg/
day).
The
lactation
index
was
also
significantly
reduced
(
94.6%)
in
the
1.6
dose
group.
In
addition,
gestation
length
was
significantly
reduced
in
the
high­
dose
group
and
there
also
was
a
significant
reduction
in
the
number
of
implantation
sites
followed
by
a
concomitant
reduction
in
litter
size.
Statistically
significant
reductions
in
pup
body
weights
were
also
observed
at
the
two
highest
dose
groups.
Other
adverse
signs
in
the
3.2
mg/
kg/
day
dose
level
associated
with
reductions
in
pup
viability
and
maternal
care
included
litters
with
pups
that
were
not
nursing
or
who
had
no
evidence
of
milk
in
the
stomach,
as
well
as
maternal
cannibalization
of
pups
that
were
stillborn
or
found
dead.
ENV/
JM/
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2002)
17/
FINAL
45
Since
F1
generation
pup
viability
was
significantly
reduced
in
the
1.6
and
3.2
mg/
kg/
day
dose
groups,
only
the
0.1
and
0.4
mg/
kg/
day
dose
groups
were
carried
into
the
second
generation.

Clinical
observations
in
the
F1
generation
male
rats
appeared
unremarkable.
No
treatment­
related
deaths
were
reported
and
no
statistically
significant
differences
were
reported
for
any
of
the
following
parameters:
body
weights/
body
weight
gains,
average
day
of
preputial
separation;
values
for
learning,
short­
term
retention,
long­
term
retention
or
response
inhibition
as
evaluated
by
performance
in
a
passive
avoidance
or
watermaze
performance
paradigm;
mating
or
fertility
parameters;
necroscopic
examinations;
absolute
or
relative
weights
for
the
right
or
left
testis,
seminal
vesicles,
right
epididymis,
or
prostate;
and
terminal
body
weights.
The
only
reported
effects
were
significant
reductions
in
absolute
food
consumption
on
postweaning
days
1­
8
occurring
at
the
0.1
and
0.4
mg/
kg/
day
dose
levels.

Clinical
observations
for
the
F1
generation
females
were
likewise
unremarkable.
Observations
at
the
0.4
mg/
kg/
day
dose
group
included,
reductions
in
body
weights
on
day
1
postweaning,
significant
losses
in
body
weight
on
LDs
1­
4,
and
significant
reductions
in
food
consumption
on
days
1­
8
postweaning
and
during
lactation.
There
were
no
statistically
significant
differences
reported
for
any
of
the
following
parameters:
values
for
learning,
short­
term
retention,
long­
term
retention
or
response
inhibition
as
evaluated
by
performance
in
a
passive
avoidance
or
water
maze
performance
paradigm;
mating
and
fertility
parameters;
gestation
index;
pregnancy
rates;
and
necroscopic
examinations.

Evidence
of
treatment­
related
effects
in
the
F2
generation
pups
consisted
of
reductions
in
mean
pup
body
weights
(
on
a
per
litter
basis)
observed
at
0.1
mg/
kg/
day
on
LD
4
and
7.
Body
weights
were
comparable
to
control
levels
by
LD
14.
At
0.4
mg/
kg/
day,
statistically
significant
reductions
in
mean
pup
body
weights
were
observed
on
LDs
7
and
14.
Mean
body
weights
on
LD21
continued
to
remain
lower
than
controls,
although
the
difference
was
not
statistically
significant
(
46.5
g
in
0.4
mg/
kg/
day
dose
group
vs.
50
g
in
controls).
Clinical
and
necroscopic
observations
of
the
F2
generation
pups
were
unremarkable.
No
other
toxicologically
significant
effects
were
reported.

Under
the
conditions
of
the
study,
the
NOAEL
and
LOAEL
for
both
the
F0
generation
male
and
female
parents
are
0.1
mg/
kg/
day
and
0.4
mg/
kg/
day,
respectively,
based
on
reductions
in
body
weight
gain
and
food
consumption.
The
NOAEL
for
the
F1
generation
parental
males
could
not
be
established
since
treatment­
related
reductions
in
absolute
food
consumption
values
were
reported
at
the
lowest
dose
tested,
0.1
mg/
kg/
day.
The
NOAEL
and
LOAEL
for
the
F1
generation
parental
females
are
0.1
mg/
kg/
day
and
0.4
mg/
kg/
day,
respectively,
based
on
significant
reductions
in
body
weights
and
food
consumption.
The
NOAEL
and
LOAEL
for
the
F1
generation
offspring
are
0.4
mg/
kg/
day
and
1.6
mg/
kg/
day,
respectively,
based
on
significant
reductions
in
the
number
of
implantation
sites,
litter
size,
pup
viability,
pup
body
weight
and
survival.
The
NOAEL
and
LOAEL
for
the
F2
generation
offspring
are
0.1
mg/
kg/
day
and
0.4
mg/
kg/
day,
respectively,
based
on
significant
reductions
in
mean
pup
body
weight.

Liver
and
sera
samples
were
collected
from
the
F0
and
F1
animals
at
terminal
sacrifice
(
after
cohabitation
for
males
and
on
lactaton
day
21
for
females)
and
analyzed
for
the
presence
of
PFOS
(
3M
Environmental
Laboratory,
1999a).
The
results
for
the
F0
animals
are
presented
below:
ENV/
JM/
RD(
2002)
17/
FINAL
46
Dose
group
Average
PFOS
conc.
Average
PFOS
conc.
(
mg/
kg/
day)
in
serum
(
ug/
ml)
in
liver
(
ug/
g)

0.0
female
0.0307
female
0.171
male
0.0244
male
0.665
0.1
female
5.28
female
14.8
male
10.5
male
84.9
0.4
female
18.9
female
58.0
male
45.4
male
176
1.6
female
82
female
184
male
152
male
323
3.2
female
NR*
female
NR*
male
273
male
1360
*
samples
not
received
Qualitatively,
the
F0
results
indicate
all
rats
(
including
controls)
had
detectable
levels
of
PFOS
in
serum
and
livers.
PFOS
concentration
increased
with
dose.
PFOS
concentrations
were
higher
in
the
liver
than
in
the
serum,
and
males
had
greatly
increased
PFOS
concentrations
in
serum
and
liver
when
compared
with
females
of
the
same
dose
group.
Pooled
liver
samples
from
the
F1
animals
sacrificed
shortly
after
birth
had
lower
PFOS
concentrations
than
adults
of
the
F0
generation
of
the
same
dose
group.
The
average
PFOS
concentrations
in
pooled
liver
samples
from
F1
animals
shortly
after
birth
were
0.0511,
6.19,
57.6,
and
70.4
ug/
g
in
the
0.0,
0.1,
0.4,
and
1.6
mg/
kg/
day
dose
groups,
respectively.
These
quantitative
values
for
the
PFOS
concentration
in
the
liver
and
serum
should
be
viewed
with
caution.
The
accuracy
of
quantitation
is
+
30%,
the
purity
of
the
analytical
reference
substance
is
unknown,
and
there
were
several
uncorrected
dilution
errors.

Two
studies
were
then
conducted
to
further
understand
the
distribution
of
PFOS
in
the
dam,
fetus
and
neonate.
In
the
first
study,
Sprague­
Dawley
rats
were
administered
oral
doses
of
PFOS
(
0.1,
0.4,
1.6,
and
3.2
mg/
kg/
day)
once
daily
beginning
42
days
prior
to
cohabitation,
and
continued
through
day
14
or
day
20
of
presumed
gestation
(
3M
Environmental
Laboratory,
2001e).
Serum,
urine,
and
feces
specimens
were
collected
from
the
dams
before
mating
and
at
GD
7,
GD
15
and
GD
21;
liver
specimens
were
collected
on
GD21.
A
total
of
54
pooled
serum
and
liver
specimens
were
collected
from
fetuses
on
GD
21.
The
results
are
shown
below:

Average
Results
for
the
Analysis
of
Serum
Samples
(
ug/
ml)

0
mg/
kg
0.1
mg/
kg
0.4
mg/
kg
1.6
mg/
kg
3.2
mg/
kg
Day
0
0.0723
8.89
40.7
160
318
Day
7
0.126
7.82
40.9
154
105
Day
15
0.0926
8.80
41.4
156
275
Day
21
0.0714
4.24
26.2
136
155
Day
21
Fetal
0.125
9.07
34.3
101
165
ENV/
JM/
RD(
2002)
17/
FINAL
47
Average
Results
for
the
Analysis
of
Liver
Samples
Dose
Group
PFOS
Conc.
(
ug/
g)
PFOS
Conc.
(
ug/
g)
(
mg/
kg/
day)
Female
Adult
Fetal
Liver
0
0.288
0.169
0.1
29.2
7.93
0.4
107
30.6
1.6
347
86.7
3.2
610
230
In
general,
there
was
a
dose­
related
increase
in
the
levels
of
PFOS
in
the
liver
and
serum
of
the
dams
and
the
fetuses.
PFOS
was
also
observed
in
the
control
dams,
as
well
as
the
control
fetuses.
As
observed
in
the
2­
generation
reproductive
toxicity
study
described
above,
the
levels
of
PFOS
were
much
higher
in
the
liver
than
in
the
serum
for
the
dams.
The
levels
of
PFOS
remained
fairly
steady
in
the
serum
of
the
dams
from
GD0
 
GD
15,
but
the
levels
dropped
at
GD21.
In
the
GD21
fetuses,
the
level
of
PFOS
in
the
serum
was
generally
comparable
to
the
level
observed
in
the
dams,
whereas
the
level
of
PFOS
in
the
fetal
livers
was
well
below
that
seen
in
the
dams.

In
the
second
study,
Sprague­
Dawley
rats
were
administered
oral
doses
of
PFOS
(
0.1
and
1.6
mg/
kg/
day),
once
daily
beginning
43
days
prior
to
cohabitation
until
confirmed
evidence
of
mating
(
3M
Environmental
Laboratory,
2001f).
Urine
and
fecal
samples
were
collected
from
the
dams
during
the
following
intervals:
one
day
prior
to
initiation
of
cohabitation
to
the
following
morning,
days
6
to
7,
14
to
15,
and
20
to
21
of
presumed
gestation
and
days
21­
22of
lactation.
Blood
samples
were
collected
from
each
of
the
dams
on
the
day
cohabitation
was
initiated
(
prior
to
cohabitation),
GD
7,
15
and
21,
and
LD
14
and
22.
Day
1
of
lactation
was
defined
as
the
day
of
birth.
On
LD
4,
litters
were
culled
to
five
male
pups
and
five
female
pups
per
litter,
where
possible.
Sera
specimens
were
collected
from
pooled
litter
samples
on
LD
21.
Liver
specimens
were
collected
from
the
dams,
and
the
liver
from
each
pup
was
collected
and
pooled
per
litter.
Blood
samples
were
collected
and
pooled
per
liter.
The
results
are
shown
below:

Average
Results
for
the
Analysis
of
Serum
Samples
(
ug/
ml)

0
mg/
kg
0.1
mg/
kg
1.6
mg/
kg
GD
0
0.100
9.21
161
GD
7
0.0796
7.24
129
GD
15
0.0742
5.68
90.6
GD
21
<
LLQ
2.58
39.5
LD14
0.0542
1.63
20.6
LD22
0.0492
0.979
14.1
LD
21
Pups
0.0531
1.80
27.1
Average
Results
for
the
Analysis
of
Liver
Samples
(
ug/
g)

0
mg/
kg
0.1
mg/
kg
1.6
mg/
kg
Dam
0.243
6.15
59.7
Pup
0.174
5.00
56.2
In
general,
there
was
a
dose­
related
increase
in
the
levels
of
PFOS
in
the
liver
and
serum
of
the
dams
and
the
levels
in
the
serum
decreased
with
time.
The
levels
of
PFOS
were
much
higher
in
the
liver
than
in
the
ENV/
JM/
RD(
2002)
17/
FINAL
48
serum
of
the
dams
and
the
pups.
The
levels
of
PFOS
were
similar
in
the
liver
of
the
dams
and
pups,
while
the
levels
in
the
serum
were
slightly
higher
in
the
pups
than
in
the
dams.

Based
on
the
results
of
the
two­
generation
reproductive
toxicity
study
in
which
significant
reductions
in
pup
viability
were
observed
at
1.6
and
3.2
mg/
kg/
day,
a
cross­
fostering
study
was
conducted
as
a
means
of
determining
whether
the
effects
observed
in
pups
were
a
result
of
in
utero
exposure
to
PFOS
or
as
a
result
of
exposure
during
lactation;
thus
the
potential
for
a
distinction
to
be
made
between
prenatal
and
postnatal
effects
following
continuous
maternal
treatment
(
Christian
et
al.,
1999c).

In
this
study,
two
groups
of
25
female
Sprague­
Dawley
rats
were
administered
0
and
1.6
mg/
kg/
day
PFOS
in
0.5%
Tween­
80
by
gavage,
beginning
42
days
prior
to
mating
to
untreated
(
breeder)
males,
and
continuing
throughout
gestation
and
into
day
21
of
lactation.
A
dose
volume
of
5
mL/
kg
was
administered,
adjusted
daily
on
the
basis
of
individual
body
weight.
Parental
females
were
observed
twice
daily
for
viability
and
clinical
observations
were
recorded
1
hour
prior
to
and
after
dosing
during
the
treatment
period.
Maternal
body
weights
were
recorded
once
during
the
acclimation
period
and
then
daily
during
the
treatment
period
and
at
sacrifice;
food
consumption
was
also
recorded
once
during
the
acclimation
period
and
then
daily
during
gestation
and
on
days
1,
4,
7,
10,
and
14
of
lactation.
During
parturition,
females
were
continually
evaluated
for
clinical
signs
and
also
for
duration
of
gestation,
length
of
parturition,
litter
sizes,
and
pup
viability
at
birth.
Maternal
behavior
was
recorded
daily
throughout
lactation.
All
maternal
rats
were
sacrificed
by
carbon
dioxide
asphyxiation
on
day
22
of
lactation
and
a
gross
necropsy
of
the
thoracic,
abdominal,
and
pelvic
viscera
was
performed;
any
gross
lesions
were
preserved
for
future
analysis.
In
addition,
the
number
and
distribution
of
implantation
sites
were
recorded.
Rats
that
did
not
deliver
a
litter
were
not
included
in
the
cross­
fostering
procedure
and
were
sacrificed
on
lactation
day
25,
examined
for
gross
lesions,
and
the
uteri
examined
to
confirm
the
presence/
absence
of
implantation
sites.
Dams
with
no
surviving
pups
were
sacrificed
after
the
last
pup
was
found
dead,
missing,
or
presumed
cannibalized.

Following
completion
of
parturition,
litters
were
immediately
removed
from
their
respective
dams
and
placed
with
either
a
control­
or
PFOS­
treated
dam
for
rearing.
This
cross­
fostering
procedure
resulted
in
four
groups
of
12­
13
dams
or
pups
as
follows:
A)
control
dams
with
litters
from
PFOS­
treated
dams,
i.
e.,
in
utero
exposure
only;
B)
control
dams
with
litters
from
control
dams,
i.
e.,
negative
control;
C)
PFOStreated
dams
with
litters
from
PFOS­
treated
dams,
i.
e.,
both
in
utero
and
post­
natal
exposure;
and
D)
PFOS­
treated
dams
with
litters
from
control
dams,
i.
e.,
post­
natal
exposure
only.

On
day
1
of
lactation
(
birth),
each
pup
was
individually
weighed
and
each
litter
was
evaluated
twice
daily
during
lactation
for
viability.
Pups
were
observed
once
daily
for
clinical
signs
and
gross
external
physical
anomalies.
Pup
body
weights
were
recorded
on
days
1,
4,
7,
14,
and
21
of
lactation,
and
then
at
sacrifice.
On
day
4
of
lactation,
each
cross­
fostered
litter
was
culled
to
5
males
and
5
females.
On
day
21
of
lactation,
all
pups
were
sacrificed
via
decapitation
and
examined
for
gross
lesions.
Pups
found
dead
or
sacrificed
because
of
moribundity
were
examined
for
gross
lesions
and
for
the
cause
of
death
or
the
moribund
condition.
The
lungs,
liver,
and
any
gross
lesions
were
collected
from
selected
pups
at
various
timepoints
and
preserved
for
possible
future
analysis.
Liver
samples
were
evaluated
via
electron
microscopy.
The
method
of
statistical
analysis
consisted
of
calculation
of
averages
and
percentages;
litter
values
were
used
where
appropriate.

In
addition,
samples
of
blood,
milk
(
including
the
milk­
secreting
glands),
and
liver
were
collected
from
selected
maternal
rats
and
pups
(
blood
and
liver
samples
were
pooled
per
litter)
at
various
timepoints
for
analysis
of
PFOS
concentration.
Only
the
analysis
of
the
sera
samples
are
available
at
this
time.

All
maternal
rats
survived
to
schedule
sacrifice.
Signs
of
clinical
toxicity
observed
in
the
dams
during
the
study
period
(
e.
g.,
chromorhinorrhea,
scaly
tail,
abrasion
on
the
head,
neck,
tail
and/
or
forelimb,
missing,
ENV/
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2002)
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broken
and/
or
misaligned
incisors,
and
localized
alopecia,
among
others)
were
not
considered
to
be
treatment­
related
since
they
also
occurred
in
the
control
animals.
Mean
maternal
body
weight
gains
at
1.6
mg/
kg/
day
were
reduced
compared
to
controls
during
premating
and
continuing
throughout
gestation.
Mean
maternal
body
weights
and
body
weight
changes
in
the
treated
group
were
comparable
to
those
seen
in
control
animals
during
lactation.
During
the
premating
period,
and
on
into
gestation
and
lactation,
food
consumption
was
reduced
in
treated
animals
as
compared
to
controls.
Reductions
in
gestation
length,
the
average
number
of
implantation
sites,
delivered
sizes,
and
live
litter
size
were
observed
in
treated
animals.
All
pregnant
animals
delivered
live
offspring.

Following
cross­
fostering
on
LD
1,
live
litter
sizes
were
comparable
between
treated
and
control
groups.
Pup
mortality
was
observed
in
two
of
the
cross­
fostered
groups.
On
lactation
days
2­
4,
approximately
19%
of
the
pups
in
group
C
were
either
found
dead
or
presumed
cannibalized.
Pup
mortality
was
also
observed
in
group
A
at
a
rate
of
9%.
In
addition,
on
day
4
of
lactation,
the
number
of
live
pups,
numbers
of
surviving
pups
per
litter,
and
live
litter
sizes
were
also
reduced
in
these
two
groups.
Pup
mortality
in
groups
B
and
D
during
lactation
days
2­
4
were
at
1.6%
and
1.1%,
respectively.
Reductions
in
pup
body
weights
were
observed
in
groups
A
and
C
on
day
1
of
lactation.
Pup
body
weights
in
group
D
were
comparable
to
controls
during
that
same
period.
From
lactation
day
4­
21,
pup
body
weights
in
groups
A,
C,
and
D
were
reduced
when
compared
to
group
B
(
negative
control),
with
the
reductions
greatest
in
group
C.
Two
litters
in
group
A
and
one
litter
on
group
C
did
not
nurse.
Milk
analysis
of
the
stomachs
of
pups
found
no
milk
in
the
stomachs
of
57%,
100%,
and
87%
of
the
pups
found
dead
and
necropsied
in
groups
A,
C,
and
D,
respectively.
Sex
ratios
and
the
lactation
index
were
comparable
among
all
groups.
Signs
of
clinical
toxicity
were
observed
in
pups,
but
were
not
considered
to
be
treatment­
related
since
they
also
occurred
in
group
B
(
negative
control)
at
the
same
rate.

Electron
microscopic
examination
of
the
liver
revealed
an
increase
in
the
number
of
peroxisomes
in
pups
from
dams
treated
with
1.6
mg/
kg/
day
PFOS.
No
significant
differences
were
observed
between
group
B
and
the
other
groups
following
examination
of
pup
lungs.

PFOS
concentrations
in
the
serum
of
untreated
dams
ranged
from
below
the
limit
of
detection
(
0.05
ug/
ml)
to
5.34
ug/
ml.
Serum
PFOS
concentrations
in
the
pups
from
untreated
dams,
fostered
with
untreated
dams,
were
below
the
limit
of
detection.
Serum
PFOS
concentrations
in
the
pups
from
treated
dams,
fostered
with
untreated
dams,
ranged
from
47.6
ug/
ml
to
59.2
ug/
ml.
PFOS
concentrations
in
the
serum
of
treated
dams
ranged
from
59.2
ug/
ml
to
157
ug/
ml.
Serum
PFOS
concentrations
in
the
pups
from
untreated
dams,
fostered
with
treated
dams,
ranged
from
below
the
limit
of
detection
to
35.7
ug/
ml.
Serum
PFOS
concentrations
in
the
pups
from
treated
dams,
fostered
with
treated
dams,
ranged
from
79.5
ug/
ml
to
96.9
ug/
ml.
These
data
indicate
that
exposure
to
PFOS
can
occur
both
in
utero
and
via
milk
from
treated
dams
(
3M
Environmental
Laboratory,
1999b).
The
accuracy
of
quantitation
for
the
analyses
was
+
30%.

In
conclusion,
pups
from
control
dams
that
were
cross­
fostered
with
PFOS­
treated
dams
(
post­
natal
exposure
only)
had
the
same
low
mortality
rate
(
1.1%)
as
pups
from
control
dams
cross­
fostered
with
control
dams
(
1.6%;
negative
control).
Mortality
rates
in
the
remaining
two
groups,
however
(
i.
e.
control
dams
with
litters
from
PFOS­
treated
dams,
i.
e.,
in
utero
exposure
only;
and
PFOS­
treated
dams
with
litters
from
PFOS­
treated
dams,
i.
e.,
both
in
utero
and
post­
natal
exposure),
were
much
higher
at
9.6
%
and
19.2%,
respectively.
Under
the
limited
conditions
of
the
study,
the
data
appear
to
indicate
that
reduced
pup
survival
is
mainly
a
result
of
in
utero
exposure
to
PFOS
and
that
post­
natal
exposure
via
milk
in
conjunction
with
in
utero
exposure
may
also
contribute
to
reduced
pup
survival.
In
contrast,
exposure
during
lactation
alone,
through
milk
from
exposed
dams,
does
not
appear
to
have
any
adverse
affect
on
pup
viability.
Additionally,
analysis
of
PFOS
concentration
showed
that
PFOS
was
observed
in
the
sera
of
F0
female
rats
exposed
during
the
in­
life
phase
of
the
study.
Additionally,
PFOS
was
observed
in
sera
samples
taken
from
F1
generation
pups
from
female
rats
exposed
to
the
test
substance,
and
in
F1
generation
pups
exposed
via
lactation,
but
not
exposed
in
utero.
ENV/
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Several
mechanistic
studies
are
being
conducted
to
understand
the
neonatal
death
(
3M
Company,
2001c).
Preliminary
results
indicate
that
reductions
in
serum
lipids
and
cholesterol
synthesis
do
not
appear
to
play
a
significant
role
in
the
death
of
the
offspring.

3.8
Human
Hazard
Several
occupational
studies
on
the
health
effects
associated
with
fluorochemical
exposure
have
been
conducted.
The
following
studies
have
all
been
conducted
at
the
3M
Decatur,
Alabama
plant
where
PFOS
has
been
manufactured
since
1991,
and
PFOA
since
1998.
The
studies
conducted
in
2000
also
included
the
Antwerp,
Belgium
plant.
Only
some
of
the
studies
examined
a
possible
association
between
serum
PFOS
levels
in
workers
and
corresponding
health
effects.
The
other
studies
use
exposure
categories
as
surrogates
for
fluorochemical
serum
levels
in
workers.

Cross­
sectional
data
from
medical
surveillance
conducted
at
the
plants
were
analyzed
to
determine
if
there
were
any
associations
between
PFOS
levels
and
hematology,
clinical
chemistries,
and
hormonal
parameters
(
Olsen
et
al.,
1998,
1999,
2001a).
A
group
of
male
volunteers
working
at
the
Decatur
plant
in
1995
and
1997
and
volunteers
from
the
2000
medical
surveillance,
which
also
included
females
and
Antwerp
employees,
were
analyzed.

In
the
1995
and
1997
analyses,
there
were
no
consistent
associations
between
PFOS
levels
in
the
workers
and
the
hematology
and
other
clinical
chemistry
parameters
when
the
data
from
both
plants
were
combined.
Mean
serum
cholesterol
levels
remained
constant
or
increased
with
increasing
PFOS
serum
levels
depending
on
the
year;
however,
those
employees
with
the
highest
PFOS
levels
had
lower
mean
HDL
values.
It
should
be
noted
that
age
was
significantly
associated
with
higher
PFOS
levels.
In
addition,
there
were
no
associations
between
clinical
hepatic
enzyme
tests
and
serum
PFOS
levels
in
workers.

Eleven
hormone
values
were
collected
from
88
of
the
employees
who
participated
in
the
study
in
1995.
The
employees
who
participated
in
the
hormone
portion
of
the
study
were
significantly
different
from
the
non­
participants
at
both
locations
(
younger,
higher
PFOS
levels,
smokers,
and
higher
WBC).
After
adjusting
for
confounders,
there
were
no
significant
associations
between
PFOS
and
the
hormones
analyzed,
except
estradiol;
however,
it
seems
that
one
employee
with
high
PFOS
measurements
(
12.83
ppm)
and
a
large
BMI
may
have
influenced
these
results.
Removal
of
this
employee
from
the
analyses
resulted
in
no
significant
associations
with
estradiol.

There
were
several
differences
between
the
employees
at
the
two
plants,
such
as
body
mass
index,
alcohol
consumption,
and
age.
In
addition,
there
were
only
61
employees
common
to
both
the
1995
and
1997
cohort
due
to
a
high
rate
of
turnover
at
both
plants
from
1996­
1997.
Some
of
the
findings
were
not
consistent
over
the
2
time
periods.
The
participation
rate
in
this
voluntary
biomonitoring
was
very
low
at
the
Decatur
plant
(
35­
40%).
In
addition,
the
employees
at
both
sites
used
APFO
as
a
surfactant
(
mean
PFOA
serum
levels
in
1995
were
1.46
ppm
and
in
1997
were
1.57
ppm).
The
number
of
years
that
each
group
worked
at
the
2
plants
(
exposure)
were
not
provided
in
the
analyses,
although
the
youngest
employees
had
the
lowest
levels
of
serum
PFOS.
Also,
ninety­
five
percent
of
the
employees
across
both
plants
and
both
time
periods
had
serum
PFOS
levels
less
than
6
ppm.
These
limitations
indicate
that
the
results
of
this
study
should
be
interpreted
carefully.

The
results
from
the
2000
cross­
sectional
analysis
were
analyzed
for
all
employees
from
both
plant
locations
combined
and
by
plant
since
the
Antwerp
and
Decatur
populations
were
significantly
different
from
each
other
in
several
ways.
Antwerp
employees
as
compared
to
Decatur
employees
had
lower
PFOS
serum
levels
(
mean
0.96
ppm),
were
younger,
had
lower
BMIs,
worked
fewer
years,
had
higher
alcohol
consumption,
higher
mean
HDL
and
bilirubin
values,
lower
mean
triglyceride,
alkaline
phosphatase,
GGT,
ENV/
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AST,
and
ALT
values,
and
mean
thyroid
hormone
values
tended
to
be
higher.
The
data
were
also
stratified
by
production
status
and
gender
and
divided
into
quartiles
of
their
serum
PFOS
distribution.
However,
the
PFOS
levels
are
different
for
each
subgroup,
making
it
difficult
to
compare
the
results.

Comparisons
across
quartiles
within
plant
locations
did
not
reveal
many
differences
in
thyroid,
hematology,
or
urinanalysis
values.
The
only
significant
(
p
<
.05)
differences
noted
were:
BUN
was
significantly
higher
in
male
production
workers
and
females
in
the
highest
PFOS
quartiles
at
the
Antwerp
plant,
and
ALT
was
significantly
higher
in
the
highest
quartile
in
Decatur
male
production
employees.

When
the
data
were
analyzed
by
number
of
employees
who
had
values
above
the
reference
range
for
hepatic
clinical
chemistry
tests
and
liver
enzyme
and
bilirubin
tests,
a
higher
percentage
of
male
Decatur
production
workers
were
in
the
highest
PFOS
quartile
(
2.31­
10.06
ppm)
for
ALT,
GGT,
and
total
liver
panel
than
the
other
quartiles.
Most
notable
were
the
results
for
ALT
where
8%
of
employees
in
the
lowest
PFOS
exposure
group
(
Q1)
and
28%
in
the
highest
exposure
group
(
Q4)
had
values
above
the
reference
range,
while
the
percentages
for
total
liver
panel
(
which
includes
alkaline
phosphatase,
AST,
ALT,
GGT,
and
total
and
direct
bilirubin)
were
18%
and
35%,
respectively.
This
trend
was
not
evident
in
Decatur
nonproduction
employees,
in
Decatur
females,
or
in
any
of
the
Antwerp
employees.
However,
it
should
be
noted
that
each
sub­
population
had
a
different
serum
PFOS
quartile
distribution.

Mean
values
for
triglycerides,
alkaline
phosphatase,
total
bilirubin,
and
ALT
were
significantly
(
p
<
.05)
higher
in
Q4
(
mean
PFOS
level
2.69,
range
1.69
 
10.06
ppm)
than
in
Q1
(
mean
PFOS
level
0.27
ppm,
range
0.04
 
0.42
ppm)
for
all
male
employees
at
both
plants
(
n
=
421).
It
should
be
noted
that
the
number
of
Antwerp
production
employees
were
evenly
distributed
among
the
quartiles
while
this
was
not
the
case
for
Decatur
employees.
The
highest
number
of
Decatur
employees
was
in
Q4.
These
data
were
analyzed
for
employees
who
had
values
above
the
reference
range
for
alkaline
phosphatase,
AST,
ALT,
GGT,
and
total
liver
panel.
For
male
employees
for
all
of
these
measures,
the
levels
increased
from
Q1
to
Q4;
however,
statistical
significance
was
not
reported.
In
Q1,
4%
of
the
employees
had
values
above
the
reference
range
for
ALT
and
6%
for
GGT,
while
12%
was
reported
for
Q4
for
both
of
these
tests.
For
total
liver
panel,
14%
of
the
employees
had
values
above
the
reference
range
in
Q1
as
compared
to
23%
in
Q4.
The
numbers
of
female
employees
with
values
above
the
reference
range
was
very
small
(
n
=
8).
In
female
employees
combined
for
both
plants
(
n
=
97),
alkaline
phosphatase
and
GGT
were
significantly
higher
(
p
<
.05)
and
total
bilirubin
significantly
lower
in
Q4
than
in
Q1.

Thyroid
results
for
male
production
employees
of
both
plants
indicated
that
T3
was
significantly
higher
(
p
<
.05)
and
THBR
(
T3
uptake)
was
significantly
lower
(
p
<
.05)
in
Q4
than
in
Q1.
The
range
of
T3
values
was
very
large
for
females;
however,
there
were
no
statistically
significant
differences
between
the
mean
values
across
the
quartiles.

Multivariable
regression
analyses
were
conducted
to
adjust
for
possible
confounders
that
may
affect
the
results
of
the
clinical
chemistry
tests.
The
following
variables
were
included:
production
job
(
yes
or
no),
plant,
age,
BMI,
cigarettes/
day,
drinks/
day
and
years
worked
at
the
plant.

A
positive
significant
(
p
=
.04)
association
between
T3
and
PFOS
was
observed.
Plant
location
was
highly
significant
(
p
<
.0001)
in
the
model.
BMI,
cigarettes/
day,
alcohol/
day
were
also
significant.
In
the
univariate
analyses,
Antwerp
employees
had
higher
mean
T3
levels
than
Decatur
employees
overall.
However,
for
each
plant
(
individually)
T3
values
increased
by
quartile
as
PFOS
serum
levels
increased,
although
the
differences
were
not
statistically
significant.
THBR,
as
well
as
the
other
thyroid
hormones,
were
not
significant
in
the
regression
analyses.

A
positive
statistically
significant
(
p
=
.04)
association
between
PFOS
and
cholesterol
was
observed.
When
both
PFOS
and
PFOA
were
included
in
the
model,
neither
were
statistically
significant
at
p=.
05.
ENV/
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2002)
17/
FINAL
52
PFOS
was
not
significantly
associated
with
HDL,
but
triglycerides
were
positively
associated
with
PFOS
(
p
=
.01).
No
significant
associations
were
observed
with
PFOS
in
relation
to
alkaline
phosphatase,
GGT,
AST
or
total
bilirubin.

A
longitudinal
analysis
of
the
above
data
was
performed
to
determine
whether
occupational
exposure
to
fluorochemicals
over
time
is
related
to
changes
in
clinical
chemistry
and
lipid
results
in
employees
of
the
Antwerp
and
Decatur
facilities
(
Olsen
et
al.,
2001b).
The
medical
surveillance
data
from
1995,
1997,
and
2000
were
analyzed
using
multivariable
regression.
The
plants
were
analyzed
using
3
subcohorts
that
included
those
who
participated
in
2
or
more
medical
exams
between
1995
and
2000.
When
male
employees
from
both
plants
were
combined,
no
statistically
significant
(
p
<
.05)
associations
were
observed
over
time
between
PFOS
and
cholesterol
or
triglycerides.
In
addition,
there
were
no
significant
associations
between
PFOS
and
changes
over
time
in
HDL,
alkaline
phosphatase,
GGT,
AST,
ALT,
total
bilirubin,
and
direct
bilirubin.

There
were
several
limitations
to
the
2000
cross­
sectional
and
longitudinal
studies
including:
1)
serum
PFOS
levels
in
these
workers
have
been
declining
over
time,
2)
PFOS
serum
levels
were
significantly
higher
at
the
Decatur
plant
than
at
the
Antwerp
plant,
3)
all
participants
were
volunteers,
4)
there
were
several
consistent
differences
in
clinical
chemistry
profiles
and
demographics
between
employees
of
the
Decatur
and
Antwerp
plants,
5)
PFOA
and
other
perfluorinated
chemicals
are
also
present
in
these
plants,
6)
plant
populations
cannot
be
compared
because
quartiles
are
different
for
each
subgroup,
and
7)
only
one
measurement
at
a
certain
point
in
time
was
collected
for
each
clinical
chemistry
test.
In
addition,
in
the
longitudinal
study
only
a
small
number
of
employees
participated
in
all
3
sampling
periods
(
24%),
different
labs
and
analytical
techniques
for
PFOS
were
used
each
year,
and
female
employees
could
not
be
analyzed
because
of
the
small
number
of
participants.

A
retrospective
cohort
mortality
study
was
performed
on
Decatur
employees
to
determine
whether
plant
employees
had
significantly
different
death
experiences
from
the
general
population
(
Mandel
and
Johnson,
1995).
There
were
1957
employees
who
had
worked
at
least
one
year
at
the
Decatur
plant
between
1961
and
1991.
Seventy­
four
deaths
were
recorded,
and
there
were
no
significantly
elevated
SMRs
for
all
major
causes
of
death
regardless
of
the
comparison
population
used
(
Alabama
state,
US,
or
Alabama
counties).
When
the
data
were
analyzed
by
job
description
(
chemical
department
vs.
film
plant
employees),
there
was
a
statistically
significant
deficit
for
all
causes
of
death
in
both
job
categories.

This
study
had
almost
complete
follow­
up
of
the
cohort
and
ascertainment
of
causes
of
death.
However,
since
the
workforce
was
relatively
young,
only
74
deaths
were
reported
for
1,951
employees.
The
number
of
women
in
the
study
was
very
small
(
4
deaths).
Additionally,
PFOS
serum
levels
were
not
examined
in
relation
to
mortality.

An
update
of
this
study
was
recently
conducted
to
follow
the
cohort
through
December
31,
1998,
thus
adding
7
years
of
follow­
up
(
Alexander,
2001).
This
cohort
consisted
of
2083
employees
who
had
worked
at
the
Decatur
plant
for
at
least
1
year.
There
were
145
deaths
reported,
almost
twice
that
reported
in
the
last
cohort.
In
this
study,
employees
were
placed
into
3
exposure
categories
based
on
their
job
descriptions:
high,
low,
and
non­
exposed.
Almost
half
of
the
employees
(
47%)
were
placed
in
the
high
exposure
group,
in
which
there
were
65
deaths.
Fourteen
percent
of
the
cohort
was
in
the
low
exposure
group
(
27
deaths),
and
39%
were
in
the
non­
exposed
group
(
53
deaths).

SMRs
were
calculated
using
state
of
Alabama
reference
data.
When
the
entire
cohort
was
analyzed,
SMRs
were
not
elevated
for
most
of
the
cancer
types
and
for
non­
malignant
causes.
SMRs
that
were
above
1
(
cancer
of
the
esophagus,
liver,
breast,
urinary
organs,
bladder,
and
skin)
were
also
elevated
when
the
cohort
was
limited
to
any
employee
ever
employed
in
a
high
exposure
job
(
except
breast
cancer).
Only
2
or
3
deaths
were
reported
for
each
of
these
cause­
specific
categories
and
were
not
statistically
significant,
ENV/
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2002)
17/
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53
except
for
bladder
cancer.

Workers
who
were
employed
in
high
exposure
jobs
were
13
times
more
likely
to
die
of
bladder
cancer
than
the
general
population
of
Alabama
(
SMR
=
12.77,
95%
CI
=
2.63
­
37.35).
This
effect
remained
when
the
data
were
analyzed
using
county
death
rates.
Three
male
employees
in
the
cohort
died
of
bladder
cancer
(
0.12
expected).
All
of
these
workers
had
been
employed
at
the
Decatur
plant
for
more
than
20
years,
and
all
of
them
had
worked
in
high
exposure
jobs
for
at
least
5
years
(
SMR
=
24.49).
In
the
previous
cohort
mortality
study,
1
bladder
cancer
death
was
reported.

Two
deaths
were
reported
for
liver
cancer.
One
was
in
the
low
exposure
group
and
one
in
the
high
exposure
group.
The
SMR
for
workers
who
were
employed
in
either
high
or
low
exposure
jobs
was
3.08
(
95%
CI
=
0.37
­
11.10).
Five
cases
of
cirrhosis
of
the
liver
were
reported
in
this
cohort,
2
in
the
high
exposure
group,
1
in
the
low
exposure
group,
and
2
in
the
non­
exposed.
The
observed
did
not
exceed
the
expected
mortality
experience
in
any
of
these
exposure
groups.

Most
of
the
same
limitations
discussed
above
for
the
original
cohort
study
apply
to
this
update.
A
larger
cohort
was
followed
in
the
update,
thus
reporting
twice
as
many
deaths;
however,
the
cohort
is
fairly
young
and
the
number
of
deaths
is
still
small
(
especially
for
females
in
all
categories).
Death
certificates
were
located
for
96%
of
the
cohort,
but
the
6
not
obtained
could
greatly
impact
the
results
of
the
analyses
since
the
number
of
deaths
in
most
of
the
sub­
cohorts
was
very
small.
In
addition
to
fluorochemicals,
workers
were
exposed
to
other
chemicals
in
the
workplace.
Biological
measurements
of
fluorochemicals
were
used
to
define
jobs
into
exposure
categories
based
on
job
descriptions;
however,
there
is
still
a
potential
for
misclassification
of
exposure.

The
high
ratio
of
observed
bladder
cancer
deaths
in
long­
time
employees
to
those
expected
in
the
general
population
is
troublesome.
Although
only
3
deaths
were
reported
in
high
exposure
jobs,
the
expected
number
in
the
general
population
was
0.23.
It
seems
unlikely
that
this
effect
would
be
due
to
chance,
given
the
magnitude
of
the
risk
estimate
(
12.77).
Many
years
of
follow
up
without
another
death
from
bladder
cancer
would
have
to
occur
before
there
would
no
longer
be
an
appreciable
risk.
Currently,
animal
studies
on
PFOS
have
not
reported
a
higher
incidence
of
bladder
cancer
tumors.
It
is
unlikely
that
tobacco
smoking
could
be
solely
responsible
for
the
excess
in
bladder
cancer
mortality,
given
the
13­
fold
increase.
Smoking
status
was
not
determined
for
this
cohort;
however,
lung
cancer
and
other
smoking­
related
cancers
were
not
elevated.
Therefore,
it
is
unlikely
that
tobacco
smoking
was
any
higher
in
the
employees
than
in
the
general
population.

Given
the
limitations
of
this
study,
it
is
unclear
whether
fluorochemicals
are
responsible
for
the
excess
of
bladder
cancer
deaths,
or
whether
other
carcinogens
may
be
present
in
the
Decatur
plant.
At
the
3M
Cottage
Grove
facility,
where
APFO
and
other
fluorochemicals
are
manufactured,
5
bladder
cancer
deaths
were
reported
(
SMR
=
1.31,
95%
CI
=
0.42
 
3.05).
Four
of
these
deaths
occurred
in
employees
who
did
not
work
primarily
in
the
chemical
division
and
therefore
were
assumed
to
have
little
exposure
to
APFO.
The
authors
of
the
Decatur
study
report
that
the
bladder
cancer
cases
worked
mostly
in
maintenance
jobs
or
at
the
incinerator
and
wastewater
treatment
plant
and
could
have
been
exposed
to
many
chemicals
in
addition
to
fluorochemicals.
It
is
important
that
follow
up
of
this
cohort
is
continued
in
order
to
gain
a
better
understanding
of
the
mortality
experience
of
workers
exposed
to
fluorochemicals.

In
order
to
gain
additional
insight
into
the
effects
of
fluorochemical
exposure
on
workers'
health,
an
"
episode
of
care"
analysis
was
undertaken
at
the
Decatur
plant
to
screen
for
morbidity
outcomes
that
may
be
associated
with
long­
term,
high
exposure
to
fluorochemicals
(
Olsen
et
al.,
2001c).
An
"
episode
of
care"
is
a
series
of
health
care
services
provided
from
the
start
of
a
particular
disease
or
condition
until
solution
or
resolution
of
that
problem.
Episodes
of
care
were
identified
in
employees'
health
claims
records
using
Clinical
Care
Groups
(
CCG)
software.
All
inpatient
and
outpatient
visits
to
health
care
providers,
ENV/
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2002)
17/
FINAL
54
procedures,
ancillary
services
and
prescription
drugs
used
in
the
diagnosis,
treatment,
and
management
of
over
400
diseases
or
conditions
were
tracked.

Episodes
of
care
were
analyzed
for
652
chemical
employees
and
659
film
plant
employees
who
worked
at
the
Decatur
plant
for
at
least
1
year
between
January
1,
1993
and
December
31,
1998.
Based
on
work
history
records,
employees
were
placed
into
different
comparison
groups:
Group
A
consisted
of
all
film
and
chemical
plant
workers;
Group
B
had
employees
who
only
worked
in
either
the
film
or
chemical
plant;
Group
C
consisted
of
employees
who
worked
in
jobs
with
high
POSF
exposures;
and
Group
D
had
employees
who
worked
in
high
exposures
in
the
chemical
plant
for
10
years
or
more
prior
to
the
onset
of
the
study.
Film
plant
employees
were
considered
to
have
little
or
no
fluorochemical
exposure,
while
chemical
plant
employees
were
assumed
to
have
the
highest
exposures.

Ratios
of
observed
to
expected
episodes
of
care
were
calculated
for
each
plant.
Expected
numbers
were
based
on
3M's
employee
population
experience
using
indirect
standardization
techniques.
A
ratio
of
the
chemical
plant's
observed
to
expected
experience
divided
by
the
film
plant's
observed
to
expected
experience
was
calculated
to
provide
a
relative
risk
ratio
for
each
episode
of
care
(
RREpC).
95%
confidence
intervals
were
calculated
for
each
RREpC.
Episodes
of
care
that
were
of
greatest
interest
were
those
which
had
been
reported
in
animal
or
epidemiologic
literature
on
PFOS
and
PFOA:
liver
and
bladder
cancer,
endocrine
disorders
involving
the
thyroid
gland
and
lipid
metabolism,
disorders
of
the
liver
and
biliary
tract,
and
reproductive
disorders.

The
only
increased
risk
of
episodes
for
these
conditions
of
a
priori
interest
were
for
neoplasms
of
the
male
reproductive
system
and
for
the
overall
category
of
cancers
and
benign
growths
(
which
included
cancer
of
the
male
reproductive
system).
There
was
an
increased
risk
of
episodes
for
the
overall
cancer
category
for
all
4
comparison
groups.
The
risk
ratio
was
greatest
in
the
group
of
employees
with
the
highest
and
longest
exposures
to
fluorochemicals
(
RREpC
=
1.6,
95%
CI
=
1.2
 
2.1).
Increased
risk
of
episodes
in
long­
time,
high­
exposure
employees
also
was
reported
for
male
reproductive
cancers
(
RREpC
=
9.7,
95%
CI
=
1.1
­
458).
It
should
be
noted
that
the
confidence
interval
is
very
wide
for
male
reproductive
cancers
and
the
sub­
category
of
prostate
cancer.
Five
episodes
of
care
were
observed
for
reproductive
cancers
in
chemical
plant
employees
(
1.8
expected),
of
which
4
were
prostate
cancers.
One
episode
of
prostate
cancer
was
observed
in
film
plant
employees
(
3.4
expected).

There
was
an
increased
risk
of
episodes
for
neoplasms
of
the
gastrointestinal
tract
in
the
high
exposure
group
(
RREpC
=
1.8,
95%
CI
=
1.2­
3.0)
and
the
long­
term
employment,
high
exposure
group
(
RREpC
=
2.9,
95%
CI
=
1.7
 
5.2).
Most
of
the
episodes
were
attributable
to
benign
colonic
polyps.
Similar
numbers
of
episodes
were
reported
in
film
and
chemical
plant
employees.

In
the
entire
cohort,
only
1
episode
of
care
was
reported
for
liver
cancer
(
0.6
expected)
and
1
for
bladder
cancer
(
1.5
expected).
Both
occurred
in
film
plant
employees.
Only
2
cases
of
cirrhosis
of
the
liver
were
observed
(
0.9
expected),
both
in
the
chemical
plant.
There
was
a
greater
risk
of
lower
urinary
tract
infections
in
chemical
plant
employees,
but
they
were
mostly
due
to
recurring
episodes
of
care
by
the
same
employees.
It
is
difficult
to
draw
any
conclusions
about
these
observations,
given
the
small
number
of
episodes
reported.

Chemical
plant
employees
in
the
high
exposure,
long­
term
employment
group
had
2
½
times
more
episodes
of
care
for
disorders
of
the
biliary
tract
than
their
counterparts
in
the
film
plant
(
RREpC
=
2.6,
95%
CI
=
1.2
­
5.5).
Eighteen
episodes
of
care
were
observed
in
chemical
plant
employees
and
14
in
film
plant
workers.
The
sub­
categories
that
influenced
this
observation
were
episodes
of
cholelithiasis
with
acute
cholecystitis
and
cholelithiasis
with
chronic
or
unspecified
cholecystitis.
Most
of
the
reported
episodes
occurred
in
chemical
plant
employees.
ENV/
JM/
RD(
2002)
17/
FINAL
55
Risk
ratios
of
episodes
of
care
for
endocrine
disorders,
which
included
sub­
categories
of
thyroid
disease,
diabetes,
hyperlipidemia,
and
other
endocrine
or
nutritional
disorders,
were
not
elevated
in
the
comparison
groups.
Conditions
which
were
not
identified
a
priori
but
which
excluded
the
null
hypothesis
in
the
95%
confidence
interval
for
the
high
exposure,
long­
term
employment
group
included:
disorders
of
the
pancreas,
cystitis,
and
lower
urinary
tract
infections.

The
results
of
this
study
should
only
be
used
for
hypothesis
generation.
Although
the
episode
of
care
design
allowed
for
a
direct
comparison
of
workers
with
similar
demographics
but
different
exposures,
there
are
many
limitations
to
this
design.
Episodes
of
care
are
reported,
not
disease
incidence;
therefore,
this
parameter
cannot
be
interpreted
in
any
other
manner.
The
data
are
difficult
to
interpret
because
a
large
RREpC
may
not
necessarily
indicate
high
risk
of
incidence
of
disease.
In
addition,
many
of
the
risk
ratios
for
episodes
of
care
had
very
wide
confidence
intervals.
The
analysis
was
limited
to
6
years.
Also,
the
utilization
of
health
care
services
may
reflect
local
medical
practice
patterns.
Individuals
may
be
counted
more
than
once
in
the
database
because
they
can
be
categorized
under
larger
or
smaller
disease
classifications.
Episodes
of
care
may
include
the
same
individual
several
times.
Not
all
employees
were
included
in
the
database,
such
as
those
on
long­
term
disability.
The
analysis
may
be
limited
by
the
software
used,
which
may
misclassify
episodes
of
care.
The
software
may
also
assign
2
different
diagnoses
to
the
same
episode.
Certain
services,
such
as
lab
procedures
may
not
have
been
reported
in
the
database.

4.0
Hazards
to
the
Environment
The
3M
Company
have
reported
ecotoxicity
test
results
on
five
PFOS
salts:
potassium,
lithium,
ammonium,
didecyldimethylammonium
and
DEA
(
diethanolamine).
The
majority
of
testing
has
however
been
carried
out
on
the
PFOS
potassium
salt.
Only
limited
data
are
available
for
the
other
salts,
but
because
the
salts
all
dissociate
instantaneously
at
neutral
pH
to
the
PFOS
ion
and
the
appropriate
counterion
the
potential
exists
for
read­
across.
For
the
purposes
of
this
review
it
is
assumed
that
the
toxicity
of
the
PFOS
anion
will
be
dominant
and
that
read­
across
of
toxicity
from
the
potassium
salt
to
the
other
salts
is
acceptable
as
a
first
approximation,
except
for
the
didecyldimethylammonium
salt.
This
is
because
these
counterions
are
unlikely
to
be
toxicologically
significant
compared
with
the
contribution
of
the
PFOS
ion.
Published
toxicity
data
relating
to,
for
example,
potassium
(
as
chloride)
to
fish
and
invertebrates
show
acute
and
chronic
effect
concentrations
to
be
>
100
mg/
l
(
Dierickx
and
Bredael­
Rozen,
1996;
Biesinger
and
Christensen,
1972).
Toxic
concentrations
of
the
PFOS
potassium
salt
reported
for
fish
and
invertebrates
in
the
3M
studies
are
however
all
well
below
100
mg/
l,
suggesting
that
the
potassium
ion
was
not
a
major
contributor
to
the
overall
toxicity
of
the
substance.
For
the
present
purpose
correction
for
the
molecular
weight
of
the
counterions
is
considered
to
be
unnecessary.
Didecyldimethylammonium
is
an
exception
due
not
only
to
its
higher
molecular
weight,
but
also
because
high
molecular
weight
amines
are
known
to
have
the
potential
for
exhibiting
significant
toxicity
to
aquatic
species.
Only
two
of
the
studies
presented
for
review
related
to
the
salt
of
this
cation
 
acute
studies
with
the
Fathead
minnow
(
Pimephales
promelas)
and
the
water
flea
(
Daphnia
magna).
Both
were
judged
to
be
unacceptable
for
assessing
PFOS
toxicity
because
of
the
use
of
an
inappropriate
method
of
test
medium
preparation.
The
result
of
the
test
with
the
fish
did
however
suggest
that
the
didecyldimethylammonium
ion
was
not
a
source
of
significant
toxicity
in
the
test
medium
whilst
the
result
of
the
test
with
the
water
flea
suggested
that
it
might
have
been.

Wherever
possible
measured
exposure
concentrations
have
been
used
as
the
basis
for
expressing
the
toxicity
of
the
test
substance.
In
some
instances
the
measured
concentrations
were
significantly
below
nominal.
In
basing
effect
concentrations
on
measured
concentrations
it
is
assumed
that
there
is
a
direct
relationship
between
PFOS
concentration
and
toxicity.

The
data
summaries
in
Tables
7
to
18
include
columns
headed
`
Study
Standard'
and
`
Comments',
in
addition
to
study
information
required
under
the
standard
OECD
column
headings.
ENV/
JM/
RD(
2002)
17/
FINAL
56
`
Study
standard'
indicates
the
overall
quality
of
the
reported
study
taking
into
account
the
technical
and
scientific
procedures
employed
and
the
information
content
of
the
test
report.
Five
standards
have
been
used:

 
Good
 
the
study
fulfils
all
requirements
in
terms
of
method,
reporting
and
interpretation.
 
Acceptable
 
the
study
is
of
an
overall
acceptable
standard
for
use
in
hazard
and
risk
assessment
although
there
are
inadequacies
in
some
elements.
 
Questionable
 
there
are
significant
inadequacies
in
elements
of
the
study
that
raise
concerns
about
its
validity.
The
results
should
only
be
considered
supportive
of
other
data.
 
Unacceptable
 
there
are
significant
inadequacies
in
elements
of
the
study
that
invalidate
the
data.
 
Unknown
 
there
is
insufficient
information
to
evaluate
the
standard
of
the
study.

The
column
headed
`
Comments'
highlights
observations
on
the
study
procedures
and
results
that
are
considered
relevant
to
the
assessment.
Reasons
why
procedures
or
results
were
considered
inappropriate
are
also
given.

The
tables
also
contain
a
column
headed
`
Study
reference
number'
that
refers
to
the
list
of
studies
in
Annex
1.
A
summary
of
the
lowest
acceptable
effect
concentrations
for
each
trophic
level
is
given
in
Annex
4.

4.1
Effects
on
Fish,
Invertebrates
and
Aquatic
plants
(
Algae
and
higher
plants)

Acute
(
short­
term)
data
were
available
from
a
total
of
31
studies
covering
three
species
of
freshwater
fish,
one
species
of
saltwater
fish,
a
fish
acclimated
to
saltwater,
two
species
of
freshwater
and
three
species
of
saltwater
invertebrate,
three
species
of
freshwater
and
one
species
of
saltwater
algae
and
one
species
of
freshwater
higher
plant.
The
studies
have
been
reviewed
and
this
has
resulted
in
21
being
considered
to
be
of
an
acceptable
standard
for
assessing
the
acute
(
short­
term)
toxic
hazard
of
PFOS
to
fish,
invertebrates
and
aquatic
plants.

Sub­
chronic/
chronic
(
prolonged/
longer­
term)
data
were
available
from
a
total
of
8
studies
covering
two
species
of
freshwater
fish,
one
species
of
fresh
and
saltwater
invertebrate
and
one
species
of
freshwater
unicellular
algae.
A
review
of
the
studies
has
resulted
in
seven
being
considered
to
be
of
an
acceptable
standard
for
assessing
the
toxic
hazard
of
PFOS
to
fish
and
invertebrates.
It
should
be
noted
that
study
number
25
with
the
Fathead
minnow
(
Pimephales
promelas)
was
considered
unacceptable
because
of
a
lack
information
in
the
report.
It
is
probable
that
this
study
was
the
same
as
that
described
more
fully
in
report
number
14.
Study
14
was
judged
to
be
acceptable.

In
addition,
flow­
through
bioconcentration
studies
have
been
conducted
in
the
bluegill
sunfish
(
Lepomis
macrochirus)
and
carp
(
Cyprinus
carpio).

The
acceptable
and
unacceptable
studies
are
identified
in
Table
6.
ENV/
JM/
RD(
2002)
17/
FINAL
57
Table
6.
Acceptable
and
unacceptable
studies
for
determining
the
acute
and
sub­
chronic/
chronic
(
prolonged/
long­
term)
toxic
hazard
of
PFOS
to
fish,
invertebrates
and
aquatic
plants
(
algae
and
higher
plants)

Endpoint
Test
species
Acceptable
study
reference
numbers
Unacceptable
study
reference
numbers
Pimephales
promelas
1,
16,
28
25,
26,
27
Acute
(
short­
term)
toxicity
to
fish
(
freshwater)
Lepomis
macrochirus
20
25
Oncorhynchus
mykiss
31,
42
25
Acute
(
short­
term)
toxicity
to
fish
(
saltwater)
Oncorhynchus
mykiss
Cyprinodon
variegatus
30
43
­

Pimephales
promelas
8,
14
25
Sub­
chronic/
chronic
toxicity
to
fish
(
freshwater)
Lepomis
macrochirus
41
­
Daphnia
magna
3,
15,
17,
29,
33
23,
25
Acute
(
short­
term)
toxicity
to
invertebrates
(
freshwater)
Unio
complamatus
5
­
Mysidopsis
bahia
4
­
Acute
(
short­
term)
toxicity
to
invertebrates
(
saltwater)
Crassostrea
virginica
7
­
Artemia
sp
32
­
Sub­
chronic/
chronic
toxicity
to
invertebrates
(
freshwater)
Daphnia
magna
9,
15
­

Sub­
chronic/
chronic
toxicity
to
invertebrates
(
saltwater)
Mysidopsis
bahia
10
­

Toxicity
to
freshwater
unicellular
algae
Selenastrum
capricornutum1
2,
13
24
Anabaena
flos­
aquae
36
­
Navicula
pelliculosa
38
­
Toxicity
to
saltwater
unicellular
algae
Skeletonema
costatum
39
­

Longer­
term
toxicity
to
freshwater
unicellular
algae
Selenastrum
capricornutum1
13
­

Toxicity
to
freshwater
higher
plants
Lemna
gibba
37
­

4.1.1
Fish
Acute
(
short­
term)
toxicity
The
results
of
the
acute
(
short­
term)
studies
are
summarized
in
Table
7.
Three
species
of
freshwater
fish
have
been
used
for
testing
­
Pimephales
promelas
(
Fathead
minnow),
Lepomis
macrochirus
(
Bluegill
sunfish)
and
Oncorhynchus
mykiss
(
Rainbow
trout).
The
end
point
assessed
in
all
the
tests
was
mortality
during
a
96­
hour
exposure
period.

P.
promelas
was
the
most
susceptible
freshwater
fish
species
in
acute
tests
with
a
lowest
96­
hour
LC50
for
PFOS
lithium
salt
of
4.7
mg/
l2.
L.
macrochirus,
and
O.
mykiss,
were
only
marginally
less
susceptible
­
a
96­
hour
LC50
value
of
7.8
mg/
l
was
determined
with
both
species
for
the
diethanolamine
(
DEA)
and
potassium
salt
respectively.
A
second
study
on
Oncorhynchus
mykiss
showed
an
LC50
of
22
mg/
l
again
using
the
potassium
salt.
A
very
high
96­
hour
LL50
value
of
approximately
200
mg/
l
determined
for
the
didecyldimethylammonium
salt
of
PFOS
was
obtained
for
P.
promelas
in
a
test
on
water
accommodated
1
This
species
has
recently
been
renamed
Pseudokirchneriella
subcapitata.
2
There
are
only
three
acute
toxicity
reports
for
Fathead
minnow
of
acceptable
or
better
quality,
so
the
most
sensitive
value
is
used
in
this
assessment.
ENV/
JM/
RD(
2002)
17/
FINAL
58
fractions
of
an
aqueous
mixture
containing
the
substance.
However,
the
actual
exposure
concentrations
of
PFOS
were
not
determined
and
could
not
be
estimated
in
this
test.

PFOS
also
exhibits
acute
toxicity
to
fish
in
saltwater.
A
96­
hour
LC50
value
of
13.7
mg/
l
has
been
determined
for
the
potassium
salt
in
a
test
with
O.
mykiss
acclimated
to
saltwater
at
a
salinity
of
30
parts
per
thousand.
The
data
show
saltwater
acclimated
O.
mykiss
to
be
of
similar
susceptibility
to
PFOS
when
compared
with
O.
mykiss
living
in
freshwater.
However,
in
the
absence
of
measured
exposure
concentrations,
it
should
be
noted
that
this
study
might
have
been
conducted
in
excess
of
the
substance's
salt
water
solubility
(
2.5
to
20
mg/
l,
depending
on
salinity
and
purity).
A
further
study
using
a
saltwater
fish,
Sheepshead
minnow
(
Cyprinodon
variegatus)
showed
no
toxicity
up
to
the
water
solubility
limit,
indicated
as
15
mg/
l
in
the
test.
Loss
of
concentration
during
the
test,
probably
due
to
sedimentation,
indicates
that
this
value
may
be
above
the
true
solubility,
however.

Sub­
chronic/
chronic
(
prolonged/
long­
term)
toxicity
The
results
of
the
sub­
chronic/
chronic
(
prolonged/
long­
term)
studies
are
summarized
in
Table
8.
Studies
have
been
carried
out
with
two
species
of
freshwater
fish
 
Pimephales
promelas
(
Fathead
minnow)
and
Lepomis
macrochirus
(
Bluegill
sunfish).
Tests
with
P.
promelas
were
designed
to
determine
concentrations
affecting
early
life­
stages
of
the
fish
over
exposure
periods
of
up
to
42
days.
Mortality
data
for
L.
macrochirus
were
obtained
from
a
bioconcentration
study
in
which
deaths
in
the
treated
and
control
groups
of
fish
were
recorded
over
the
62­
day
uptake
phase
of
the
study.

The
lowest
definitive
no
observed
effect
concentration
(
NOEC)
of
0.3
mg/
l
was
determined
for
P.
promelas
for
the
potassium
salt
of
PFOS.
This
value
was
applicable
to
both
survival
and
growth
end
points.
This
NOEC
is
supported
by
results
from
a
bioconcentration
study
with
L.
macrochirus
that
showed
no
significant
mortality
at
an
exposure
concentration
of
0.086
mg/
l
over
a
62­
day
uptake
phase
but
100%
mortality
at
a
concentration
of
0.87
mg/
l
after
35
days.
A
NOEC
of
1.0
mg/
l
reported
for
effects
of
PFOS
potassium
salt
on
early
life­
stages
of
P.
promelas
could
not
be
attributed
to
a
more
specific
end
point
because
insufficient
information
was
contained
in
the
study
report.

Bioconcentration
A
flow­
through
bioconcentration
study
of
PFOS
in
bluegill
sunfish
was
conducted.
Test
concentrations
of
0,
0.086
and
0.87
mg/
L
were
used.
At
the
0.086
mg/
L
level,
the
fish
were
exposed
for
62
days,
while
at
the
0.87
mg/
L
level,
the
fish
were
exposed
for
35
days
due
to
excessive
mortality.
Fish
were
collected
from
the
test
chambers
by
random
selection
at
12
time
points
during
the
study.
They
were
euthanized,
blotted
dry,
weighed
and
measured.
They
were
dissected
into
edible
and
nonedible
tissue
fractions
and
the
fractions
were
weighed.
The
head,
fins
and
viscera
were
considered
to
be
nonedible
tissue
and
the
remaining
tissue,
including
skin,
was
considered
to
be
edible
tissue.
Whole
fish
concentrations
were
calculated
from
the
sum
of
the
edible
and
nonedible
parts.
Steady­
state
BCF
values
were
calculated
from
the
tissue
concentrations
at
apparent
steady­
state
divided
by
the
mean
water
concentration.
Tissue
concentrations
were
considered
to
be
at
steady­
state
if
3
or
more
consecutive
sets
of
tissue
concentrations
were
not
significantly
different.
The
kinetic
bioconcentration
factor
(
BCFK),
uptake
rate
and
depuration
rate
were
calculated
for
the
edible,
nonedible
and
whole
fish
exposed
to
0.086
mg/
L
using
BIOFAC
computer
software.

In
this
study,
PFOS
bioconcentrated
in
the
tissues
of
bluegill
sunfish.
Apparent
steady­
state
was
not
clearly
attained
for
the
fish
exposed
to
0.086
mg
/
L.
Although
the
tissue
residue
levels
of
PFOS
were
not
statistically
significantly
different
on
days
49,
56
and
62,
the
concentration
of
PFOS
appeared
to
be
still
increasing
up
to
the
last
day
of
exposure.
BCFK
values
for
edible,
nonedible
and
whole
fish
tissues
were
calculated
to
be
1124,
4013,
and
2796,
respectively.
PFOS
depurated
slowly.
The
BIOFAC
estimates
for
ENV/
JM/
RD(
2002)
17/
FINAL
59
the
time
to
reach
50%
clearance
for
edible,
nonedible,
and
whole
fish
tissues
were
86,
116,
and
112
days,
respectively.

Kurume
Laboratory
(
2001)
conducted
a
flow­
through
bioconcentration
study
of
PFOS
in
carp
(
Cyprinus
carpio).
The
fish
were
checked
visually
and
those
demonstrating
any
abnormality
were
removed.
The
fish
were
reared
for
8
days
in
a
flow
thorough
system
following
an
external
disinfection.
After
rearing,
the
fish
were
medicated
to
eliminate
parasites
and
transferred
to
an
acclimatizing
aquarium.
After
the
second
external
disinfection,
they
were
acclimatized.
The
fish
demonstrating
any
abnormality
during
this
period
were
removed
and
the
remainder
of
the
fish
were
reared
for
15
days
in
a
flow
through
system
at
temperatures
of
25
±
2o
C.
The
fish
were
transferred
to
test
tanks
and
reared
at
the
same
temperature
in
the
flow
through
system
for
another
27
days.
Temperature
of
the
test
water
was
measured
with
alcohol
a
thermometer
and
recorded
once
a
day.
Dissolved
oxygen
concentrations
was
measured
with
a
dissolved
oxygen
probe
and
recorded
twice
a
week.
The
pH
of
the
test
water
was
measured
with
a
pH
meter
one
a
week.
Temperature
of
test
water
was
measured
and
recorded
once
a
day.
Dissolved
oxygen
in
test
water
was
measured
and
recorded
twice
a
week.
During
the
experimental
period,
the
excreta
of
carp,
dirt
on
test
tanks,
were
removed
once
a
day.
The
temperature
ranged
from
25.0
to
25.4
C;
the
dissolved
oxygen
concentration
ranged
from
7.9
to
8.1
mg/
L;
and
the
pH
ranged
from
7.6
to
7.8.

Forty
carp
were
exposed
to
two
concentrations
of
PFOS,
2
and
20
ug/
L
respectively
for
58
days
in
a
flow
though
system.
The
test
water
of
each
level
was
analyzed
once
before
firs
analysis
of
test
fish
and
at
the
same
time
as
analysis
of
test
fish.
Analysis
of
test
fish
was
performed
six
times
at
each
level
in
duration
of
exposure.
Four
fish
were
taken
out
at
each
sampling
time
and
divided
into
two
groups,
and
then
both
were
analyzed
individually.
Because
the
stored
sample
from
one
fish
was
too
small
for
the
measurement
of
lipid
content,
groups
of
two
fish
were
used.
Analysis
of
control
fish
was
performed
before
the
experimental
starting
and
after
the
experimental
completion.
Six
fish
were
taken
out
at
each
sampling
time
and
divided
into
three
groups,
and
then
both
were
analyzed
individually.

Analysis
of
PFOS
in
the
test
water
and
carp
was
performed
using
high­
performance
liquid
chromatography­
mass
spectrometry
(
HPLC­
MS)
analysis.
The
test
water
of
each
level
was
analyzed
once
before
first
analysis
of
test
fish
and
at
the
same
time
as
the
analysis
of
the
test
fish.
Steady­
state
was
reached
when
three
successive
analyses
of
BCFs
made
on
samples
taken
at
intervals
of
at
least
48
hours
were
within
+
20%
of
each
other.
When
BCFs
were
less
that
100,
it
was
evaluated
that
a
steady­
state
had
been
reached
after
28
days.
The
fish
which
were
exposed
for
61
days
were
separated
into
parts;
tegument,
head,
viscera
except
liver,
liver
and
remaining
matter.
BCFs
were
determined
in
all
the
parts.

In
this
study,
PFOS
bioconcentrated
in
the
tissues
of
carp.
Test
concentrations
of
2
and
20
ug/
L
were
used.
The
fish
were
exposed
for
58
days
to
makeup
for
excessive
mortality.
Bioconcentration
factors
were
calculated
to
be
720
for
20
ug/
L
and
200
­
1500
for
2
ug/
L.
ENV/
JM/
RD(
2002)
17/
FINAL
60
Table
7.
Acute
(
short­
term
exposure)
toxicity
data
for
PFOS
determined
in
tests
with
fish
Medium
Species
Protocol
Results
(
mg/
l)
Study
Standard
Comments
Study
Ref.

No.

Freshwater
Pimephales
promelas
(
Fathead
minnow)
OECD
203
&

OPPTS
850.1075
(
Static)
96­
hour
LC50
=
9.5
96­
hour
NOEC
=
3.3
Good
Test
substance
was
PFOS
potassium
salt.

Measured
exposure
concentrations
1
Not
noted
(
Static)
96­
hour
LC50
=
4.7
(
see
comments)
Acceptable
Test
substance
was
PFOS
lithium
salt.

Nominal
exposure
concentrations
expressed
relative
to
concentration
of
test
substance
(
24.5%
PFOS
Li
salt
and
74.5%
water).

Test
result
divided
by
4
to
express
96
h
LC50
in
terms
of
PFOS
Li
salt
concentration.

Very
little
information
on
study
protocol;

however,
result
agrees
reasonably
well
with
more
other
studies
(
1,
25,
26
and
27)
16
Not
noted
(
Static)
96­
hour
LC50
=
37.6
and
51
Unknown
Test
substance
was
PFOS
potassium
salt.

Nominal
exposure
concentrations
Standard
of
studies
cannot
be
judged
from
information
supplied.

However,
results
are
consistent
with
other
studies
(
1,
16,
26
and
27)
25
Not
noted
(
Static)
96­
hour
LC50
=
21
Questionable
Test
substance
was
PFOS
ammonium
salt.

Nominal
exposure
concentrations
Isopropanol
also
present
in
test
samples.

However,
results
are
consistent
with
other
studies
(
1,
16,
25
and
27)
26
Not
noted
(
Static)
96­
hour
LC50
=
25
Questionable
Test
substance
was
PFOS
ammonium
salt.

Nominal
exposure
concentrations
Isopropanol
also
present
in
test
samples.

Results
consistent
with
other
studies
(
1,
16,

25
and
26)
27
ENV/
JM/
RD(
2002)
17/
FINAL
61
Table
7
contd.
Acute
(
short­
term
exposure)
toxicity
data
for
PFOS
determined
in
tests
with
fish
Medium
Species
Protocol
Results
(
mg/
l)
Study
Standard
Comments
Study
Ref.

No.

Freshwater
Pimephales
promelas
(
Fathead
minnow)
OECD
203
(
Static)
96­
hour
LL50
=
200*

96­
hour
NOEL
=

<
170*

(*
calculated
assuming
35%
of
substance
in
mixture)
Acceptable
Test
substance
was
a
mixture
of
didecyldimethyammonium
salt
of
PFOS
(
approximately
35%)
and
water
with
up
to
5%
residual
perfluorochemicals.

Test
media
were
water­
accommodated
fractions
(
WAFs).

Nominal
exposure
concentrations.
28
OECD
203
&

OPPTS
850.1075
(
Static)
96­
hour
LC50
=
7.8
96­
hour
NOEC
=
4.5
(
see
comments)
Acceptable
Test
substance
was
PFOS
DEA
salt.

Nominal
exposure
concentrations
expressed
relative
to
concentration
of
test
substance
(~
25%
PFOS
DEA
salt
and
75%
water).

Results
have
therefore
been
divided
by
4
to
calculate
the
effect
concentrations
in
terms
of
PFOS
DEA
salt
concentration
based
on
reported
96­
hour
LC50
of
31
mg/
l
and
NOEC
of
18
mg/
l.
20
Lepomis
macrochirus
(
Bluegill
sunfish)
Not
noted
(
Static)
96­
hour
LC50
=
68
Unknown
Test
substance
was
PFOS
potassium
salt.

Nominal
exposure
concentrations
Standard
of
study
cannot
be
judged
from
information
supplied.
25
Oncorhynchus
mykiss
(
Rainbow
trout)
Not
noted
(
Static)
96­
hour
LC50
=
11
Unknown
Test
substance
was
PFOS
potassium
salt.

Nominal
exposure
concentrations
Standard
of
study
cannot
be
judged
from
information
supplied.
25
Standard
procedures
for
testing
acute
lethality
of
liquid
effluents
(
Environment
Canada)
96­
hour
LC50
=
7.8
Acceptable
Test
substance
was
PFOS
potassium
salt.

Sample
purity
not
characterised
Nominal
exposure
concentrations
31
ENV/
JM/
RD(
2002)
17/
FINAL
62
Table
7
contd.
Acute
(
short­
term
exposure)
toxicity
data
for
PFOS
determined
in
tests
with
fish
Medium
Species
Protocol
Results
(
mg/
l)
Study
Standard
Comments
Study
Ref.

No.

Freshwater
Oncorhynchus
mykiss
(
Rainbow
trout)
OPPTS
850.1075
OECD
203
Static
96­
hour
LC50
=
22
Acceptable
Test
substance
was
PFOS
potassium
salt.

Sample
purity
86.7%;
results
reported
as
a.
i
Based
on
measured
concentrations
42
Saltwater
Cyprinodon
variegatus
(
Sheepshead
minnow)
OPPTS
850.1075
OECD
203
Semi­
static
(
24
hour
renewal)
96­
hour
LC50
>
15
mg/
l
Acceptable
Test
substance
was
PFOS
potassium
salt.

Sample
purity
86.7%;
results
reported
as
a.
i
Based
on
mean
of
measured
concentrations
Concentration
losses
during
each
24
hour
period
would
suggest
some
sedimentation
indicating
that
the
water
solubility
may
have
been
exceeded.
Final
concentrations
in
each
24
hour
period
varied
between
11
 
16
mg/
l
43
Saltwater
Oncorhynchus
mykiss
(
Rainbow
trout)
Standard
procedures
for
testing
acute
lethality
of
liquid
effluents
(
Environment
Canada)
96­
hour
LC50
=
13.7
Acceptable
Test
substance
was
PFOS
potassium
salt.

Sample
purity
not
characterised
Test
fish
were
acclimated
to
30
parts
per
thousand
saltwater
Nominal
exposure
concentrations
In
the
absence
of
measured
exposure
concentrations,
it
should
be
noted
that
this
study
might
have
been
conducted
in
excess
of
the
substance's
salt
water
solubility
(
2.5
to
20
mg/
l,
depending
on
salinity
and
purity).
30
ENV/
JM/
RD(
2002)
17/
FINAL
63
Table
8.
Sub­
chronic/
chronic
(
prolonged/
long­
term)
toxicity
data
for
PFOS
determined
in
tests
with
fish
Medium
Species
Protocol
Results
(
mg/
l)
Study
Standard
Comments
Study
Ref.

No.

Freshwater
OECD
210
&

OPPTS
850.1400
(
Flowthrough
42­
day
NOECsurv.
=

0.30
42­
day
NOECgrowth
=

0.30
5­
day
NOEChatch
>
4.6
Good
Test
substance
was
PFOS
potassium
salt.

Measured
exposure
concentrations.

Lighting
regime
not
described
in
summary.

Conclusion
(
post­
hatch
survival
as
the
most
sensitive
endpoint):
it
is
possible
that
growth
was
also
affected
at
this
concentration,
but
the
test
design
did
not
enable
this
to
be
determined.
8
Non­
standard
(
Flowthrough
30­
day
NOECearly
lifestages
=
1
Acceptable
Test
substance
was
PFOS
potassium
salt.

Measured
exposure
concentrations.

Results
agree
with
later
study
Acetone
present
at
43
µ
l/
l
14
Pimephales
promelas
(
Fathead
minnow)
Not
noted
(
Flowthrough
NOECearly
life­
stages
=
1
Unknown
Test
substance
was
PFOS
potassium
salt.

Standard
of
study
cannot
be
judged
from
information
supplied.
It
is
however
assumed
that
this
study
is
the
same
as
14
25
(
Assumed
to
be
same
study
as
14)

Lepomis
macrochirus
(
Bluegill
sunfish)
OECD
305
&

OPPTS
850.1730
62­
day
NOECmortality
=

>
0.086,
<
0.87
Good
Bioconcentration
study
Test
substance
was
PFOS
potassium
salt.

Measured
exposure
concentrations.
41
ENV/
JM/
RD(
2002)
17/
FINAL
64
4.1.2
Invertebrates
The
results
of
acute
(
short­
term)
and
sub­
chronic/
chronic
(
prolonged/
long­
term)
studies
are
summarised
in
Tables
9
and
10.

Acute
(
short­
term)
toxicity
to
freshwater
species
Two
species
of
freshwater
invertebrate
have
been
used
for
testing
­
Daphnia
magna
(
the
`
water
flea')
and
Unio
complamatus
(
fresh
water
mussel).
Immobilisation/
mortality
during
48­
hours
exposure
was
assessed
in
the
tests
with
D.
magna.
Mortality
during
96­
hours
exposure
was
assessed
in
the
test
with
U.
complamatus.

A
lowest
acceptable
48­
hour
EC50
value
of
27
mg/
l
was
determined
for
PFOS
potassium
salt
in
tests
with
the
daphnid.
1
A
48­
hour
EL50
value
of
approximately
4.0
mg/
l
was
also
determined
for
the
didecyldimethylammonium
salt
of
PFOS
in
a
daphnid
test
on
water
accommodated
fractions
of
an
aqueous
mixture
containing
the
substance.
However,
the
actual
exposure
concentrations
of
PFOS
were
not
determined
in
this
test
and,
as
pointed
out
in
the
introduction
to
this
section,
it
is
possible
that
didecyldimethylammonium
may
have
contributed
to
the
toxicity
of
the
test
medium.
These
values
compare
with
a
96­
hour
LC50
value
determined
for
mortality
of
the
mussel
of
59
mg/
l.

Acute
(
short­
term)
toxicity
to
saltwater
species
Three
species
of
saltwater
invertebrate
have
been
used
for
testing
­
Mysidopsis
bahia
(
Mysid
shrimp),
Crassostrea
virginica
(
Eastern
oyster)
and
Artemia
sp
(
Brine
shrimp).
Mortality
during
96­
hours
exposure
was
assessed
in
the
tests
with
M.
bahia
and
Artemia
sp.
Reduction
in
shell
deposition
was
assessed
over
a
96­
hour
exposure
period
in
the
test
with
C.
virginica.
The
test
substance
was
PFOS
potassium
salt
in
all
cases.

A
96­
hour
LC50
value
of
3.6
mg/
l
and
an
associated
NOEC
of
1.1
mg/
l
were
determined
in
the
test
with
the
Mysid
shrimp.
This
compares
with
a
96­
hour
EC50
value
of
>
3.0
mg/
l
for
effects
on
shell
deposition
in
the
oyster
and
a
48­
hour
LC50
of
8.9
mg/
l
for
mortality
of
the
Brine
shrimp.

Sub­
chronic/
chronic
(
prolonged/
long­
term)
toxicity
to
freshwater
species
Two
tests
have
been
carried
out
on
PFOS
potassium
salt
with
one
species
of
freshwater
invertebrate
 
the
`
water
flea',
Daphnia
magna.
The
end
points
assessed
in
the
tests
were
survival,
growth
and
reproduction
measured
over
exposure
periods
of
up
to
28
days.

NOECs
of
12
and
7
mg/
l
have
been
determined
for
D.
magna
reproduction
in
21
and
28­
day
tests
respectively.
In
the
21­
day
test
the
NOECs
for
survival
and
growth
were
also
12
mg/
l,
indicating
that
reproduction
was
no
more
sensitive
than
these
two
other
end
points.

1
For
Daphnia
magna
there
are
five
48­
hour
acute
toxicity
studies
of
acceptable
or
better
quality,
although
one
of
these
relates
to
water
accommodated
fractions.
The
remaining
48­
hour
EC50s
are
27,
58,
61
and
210
mg/
l.
The
latter
value
is
based
on
nominal
exposure
concentrations,
but
since
the
stock
solution
was
prepared
at
a
concentration
that
exceeded
water
solubility,
exposure
concentrations
are
likely
to
be
lower
than
nominal.
This
leaves
just
three
reports
with
comparable
measures
of
toxicity,
and
so
the
most
sensitive
value
is
used
in
this
assessment.
ENV/
JM/
RD(
2002)
17/
FINAL
65
Sub­
chronic/
chronic
(
prolonged/
long­
term)
toxicity
to
saltwater
species
A
test
has
been
carried
out
on
PFOS
potassium
salt
with
one
species
of
saltwater
invertebrate;
the
Mysid
shrimp,
Mysidopsis
bahia.
The
end
points
assessed
in
the
test
were
survival,
growth
and
reproduction
measured
over
an
exposure
period
of
35
days.

The
35­
day
NOECs
determined
for
survival,
growth
and
reproduction
in
this
test
were
0.55,
0.25
and
0.25
mg/
l
respectively.
ENV/
JM/
RD(
2002)
17/
FINAL
66
Table
9.
Acute
(
short­
term
exposure)
toxicity
data
for
PFOS
determined
in
tests
with
invertebrates
Medium
Species
Protocol
Results
(
mg/
l)
Study
Standard
Comments
Study
Ref.
No.

Freshwater
Daphnia
magna
(
Water
flea)
OECD
202
&

OPPTS
850.1010
(
Static)
48­
hour
EC50
=
61
48­
hour
NOEC
=
33
Good
Test
substance
was
PFOS
potassium
salt.

Measured
exposure
concentrations
3
ASTM
1981
&

OECD
1981
(
Static)
48­
hour
EC50
=
27
Acceptable
Test
substance
was
PFOS
potassium
salt.

Nominal
exposure
concentrations.

Result
agrees
reasonably
well
with
those
obtained
in
studies
3
and
23.
15
Not
noted
(
Static)
48­
hour
EC50
=
210
48­
hour
NOEC
=
100
Acceptable
Test
substance
was
PFOS
lithium
salt.

Nominal
exposure
concentrations.

EC50
is
somewhat
higher
than
previous
value
but
stock
solution
was
prepared
at
a
concentration
that
exceeded
water
solubility,

therefore
exposure
concentrations
are
likely
to
be
lower
than
nominal.
17
OECD
202
(
Static)
24­
hour
EC50
=
>
42
48­
hour
EC50
=
14
Questionable
Test
substance
was
PFOS
potassium
salt.

Nominal
exposure
concentrations.

Data
questionable
in
view
of
the
presence
of
diethylene
glycol
butyl
ether
in
the
test
material
(
as
part
of
the
formulation).

Result
is
in
reasonable
agreement
with
later
study
(
3).
23
Not
noted
(
Static)
48­
hour
EC50
=
49.2
Unknown
Test
substance
was
PFOS
potassium
salt.

Nominal
exposure
concentrations.

Standard
of
study
cannot
be
judged
from
information
supplied.
25
ENV/
JM/
RD(
2002)
17/
FINAL
67
Table
9
contd.
Acute
(
short­
term
exposure)
and
sub­
chronic/
chronic
(
prolonged/
long­
term)
toxicity
data
for
PFOS
determined
in
tests
with
invertebrates
Medium
Species
Protocol
Results
(
mg/
l)
Study
Standard
Comments
Study
Ref.
No.

Daphnia
magna
(
Water
flea)
OECD
202
(
Static)
48­
hour
EL50
=
4.0
48­
hour
NOEL
=
2.2
Acceptable
Test
substance
was
a
mixture
of
didecyldimethylammonium
salt
of
PFOS
(
approximately
35%)
and
water
with
up
to
5%

residual
perfluorochemicals.

Test
media
were
water­
accommodated
fractions
(
WAFs).

Nominal
exposure
concentrations.

48­
hour
EL50
and
NOEL
calculated
assuming
35%
of
substance
in
mixture.
29
ISO,
1982
48­
hour
EC50
=
58
Acceptable
Test
substance
was
PFOS
potassium
salt.

Sample
purity
not
characterised
Nominal
exposure
concentrations.
33
Unio
complamatus
(
Freshwater
mussel)
OECD
203,

OPPTS
850.1075
&

ASTM­
E­
729­

88a
(
Semistatic
96­
hour
LC50
=
59
96­
hour
NOEC
=
20
Good
Test
substance
was
PFOS
potassium
salt.

Measured
exposure
concentrations.
5
Saltwater
Mysidopsis
bahia
(
Mysid
shrimp)
OPPTS
850.1035
(
Static)
96­
hour
LC50
=
3.6
96­
hour
NOEC
=
1.1
Good
Test
substance
was
PFOS
potassium
salt.

Measured
exposure
concentrations.
7
Crassostrea
virginica
(
Eastern
oyster)
OPPTS
850.1025
(
Static)
96­
hour
EC50
=
>
3.0
96­
hour
NOEC
=
1.9
Good
Test
substance
was
PFOS
potassium
salt.

Measured
exposure
concentrations.

Solubility
in
seawater
should
have
been
used
to
set
highest
test
concentration.
4
Artemia
sp.

(
Brine
shrimp)
Draft
ISO,

1981
48­
hour
LC50
=
8.9
Acceptable
Test
substance
was
PFOS
potassium
salt.

Sample
purity
not
characterised
Nominal
exposure
concentrations.
32
ENV/
JM/
RD(
2002)
17/
FINAL
68
Table
10.
Sub­
chronic/
chronic
(
prolonged/
long­
term)
toxicity
data
for
PFOS
determined
in
tests
with
invertebrates
Medium
Species
Protocol
Results
(
mg/
l)
Study
Standard
Comments
Study
Ref.
No.

Freshwater
Daphnia
magna
(
Water
flea)
OECD
211,

OPPTS
850.1300
&

ASTM
1193­

87E
(
Semistatic
21­
day
NOECrepro
=
12
21­
day
NOECsurv.
=
12
21­
day
NOECgrowth
=
12
Good
Test
substance
was
PFOS
potassium
salt.

Measured
exposure
concentrations.

No
effects
on
reproduction
at
highest
concentration
that
had
no
effect
on
mortality.
9
ASTM
1981
&

OECD
1981
(
Semi­
static)
21­
day
EC50repro.
=
12
28­
day
NOECrepro
=
7
28­
day
EC50repro.
=
11
Acceptable
Test
substance
was
PFOS
potassium
salt.

Nominal
exposure
concentrations.

Result
agrees
well
with
that
obtained
in
study
9.
15
Saltwater
Mysidopsis

bahia
(
Mysid
shrimp)
OPPTS
850.1350
(
Flowthrough
35­
day
NOECrepro.
=
0.25
35­
day
NOECsurv.
=
0.55
35­
day
NOECgrowth
=

0.25
Good
Test
substance
was
PFOS
potassium
salt.

Measured
exposure
concentrations.
10
ENV/
JM/
RD(
2002)
17/
FINAL
69
4.1.3
Aquatic
plants
The
results
of
all
the
short­
and
longer­
term
studies
are
summarised
in
Tables
11
and
12
(
algae)
and
13
(
higher
plants).
Note:
The
species
Selenastrum
capricornutum
has
been
renamed
Pseudokirchneriella
subcapitata.
For
transparency,
the
name
used
in
the
test
report
has
been
retained
in
the
discussion
below.

Freshwater
algae
 
Short­
term
Three
species
of
freshwater
unicellular
algae
have
been
used
for
testing
­
Selenastrum
capricornutum,
Anabaena
flos­
aquae
and
Navicula
pelliculosa.
The
end
point
assessed
in
the
tests
was
growth
measured
in
terms
of
cell
density,
growth
rate
and/
or
the
area
under
the
growth
curve
over
96­
hours.
Only
studies
that
assessed
toxicity
by
reference
to
effects
on
growth
rate
are
considered
here
for
the
purposes
of
determining
the
toxicity
of
PFOS
to
algae.
The
results
of
study
number
13
are
therefore
excluded
because
they
are
only
expressed
relative
to
cell
numbers
and
cell
dry
weight.
These
results
could
be
considered
were
the
EC50
values
to
be
recalculated
in
respect
of
growth
rate,
but
since
algae
were
relatively
insensitive
compared
with
fish,
this
is
not
considered
a
high
priority.

The
lowest
96­
hour
EC50
value
for
effects
on
growth
rate
of
71
mg/
l
was
determined
for
S.
capricornutum.
However
there
are
some
uncertainties
over
the
validity
of
this
result
given
that
exposure
concentrations
were
not
measured
and
diethylene
glycol
butyl
ether
was
present
in
the
test
material
(
as
part
of
the
formulation).
A
96­
hour
EC50
value
of
126
mg/
l
was
obtained
for
the
same
species
using
a
test
protocol
that
was
judged
to
meet
all
the
criteria
for
acceptability
of
the
data.
The
96­
hour
NOECs
associated
with
these
two
tests
were
35
and
44
mg/
l
respectively.
The
tests
with
the
two
other
species
yielded
96­
hour
EC50
values
of
176
mg/
l
(
A.
flos­
aquae)
and
305
mg/
l
(
N.
pelliculosa)
and
respective
NOECs
of
94
and
206
mg/
l.

Saltwater
algae
 
Short­
term
A
96­
hour
growth
inhibition
test
has
been
carried
out
on
PFOS
potassium
salt
with
Skeletonema
costatum.
The
test
was
unable
to
determine
a
definitive
96­
hour
EC50
value
because
no
effects
were
determined
at
the
highest
dissolved
PFOS
concentration
that
could
be
attained
under
the
test
conditions
(
3.2
mg/
l).

Freshwater
algae
 
Longer­
term
One
species
of
freshwater
unicellular
algae
has
been
tested
­
Selenastrum
capricornutum
(
study
reference
no.
13).
The
test
assessed
effects
on
growth
rate
expressed
relative
to
cell
density
over
14
days.
The
results
from
the
test
are
not
discussed
further
here
for
two
reasons.
Firstly,
the
results
are
expressed
relative
to
cell
density
(
see
comments
in
respect
of
short­
term
tests).
Secondly,
there
are
concerns
that
the
14­
day
test
period
and
absence
of
test
medium
renewal
may
have
resulted
in
a
decline
in
exposure
concentrations
over
time
(
the
test
media
were
not
analysed
for
PFOS
concentration).
The
latter
is
suggested
by
comparison
of
the
14­
day
EC50
value
of
95
mg/
l
with
the
96­
h
EC50
value
of
82
mg/
l
determined
in
the
same
test.

Freshwater
higher
plants
A
growth
inhibition
test
has
been
carried
out
on
PFOS
potassium
salt
with
Lemna
gibba
(
Duckweed).
The
test
yielded
a
7­
day
IC50
of
108
mg/
l
for
inhibition
of
frond
production
and
a
7­
day
NOEC
of
15.1
mg/
l
based
on
the
inhibition
of
frond
production
and
evidence
of
sub­
lethal
effects.
ENV/
JM/
RD(
2002)
17/
FINAL
70
Table
11.
Acute
(
short­
term
exposure)
toxicity
data
for
PFOS
determined
in
tests
with
algae
Medium
Species
Protocol
Results
(
mg/
l)
Study
Standard
Comments
Study
Ref.

No.

Freshwater
Selenastrum
capricornutum
OECD
201,

OPPTS
850.5400
&

ASTM
1218­
90E
(
Static)
96­
hour
EC50(
cell
density)
=
71
96­
hour
EbC50(
area
under
the
curve)
=

71
96­
hour
ErC50(
growth
rate)
=
126
96­
hour
NOEC(
growth
rate,
cell
density,

area
under
the
growth
curve)
=
44
72­
hour
EC50(
cell
density)
=
70
72­
hour
EbC50(
area
under
the
curve)
=

74
72­
hour
ErC50(
growth
rate)
=
120
72­
hour
NOEC(
growth
rate,
cell
density,

area
under
the
growth
curve)
=
70
Good
Test
substance
was
PFOS
potassium
salt.

Measured
exposure
concentrations.
2
OECD
201,

US
EPA
600/
9­
78­

018
&
ASTM­
E­

35.23
(
Static)
96­
hour
EC50(
cell
density)
=
82
96­
hour
EC10(
cell
density)
=
10
Acceptable
Test
substance
was
PFOS
potassium
salt.

Result
not
expressed
relative
to
growth
rate
Nominal
exposure
concentrations.
Result
agrees
with
those
obtained
in
studies
2
and
24.
13
OECD
201
(
Static)
96­
hour
EC50(
growth
rate)
=
71
96­
hour
NOEC(
growth
rate)
=
35
Questionable
Test
substance
was
PFOS
potassium
salt.

Nominal
exposure
concentrations.

Data
questionable
in
view
of
the
presence
of
diethylene
glycol
butyl
ether
in
the
test
material
(
as
part
of
the
formulation).
24
ENV/
JM/
RD(
2002)
17/
FINAL
71
Anabaena
flosaquae
OPPTS
850.5400
96­
hour
EC50(
growth
rate)
=
176
96­
hour
NOEC(
growth
rate)
=
94
Good
Test
substance
was
PFOS
potassium
salt.

Measured
exposure
concentrations.
36
Navicula
pelliculosa
OPPTS
850.5400
96­
hour
EC50(
growth
rate)
=
305
96­
hour
NOEC(
growth
rate)
=
206
Good
Test
substance
was
PFOS
potassium
salt.

Measured
exposure
concentrations.
38
Saltwater
Skeletonema
costatum
OPPTS
850.5400
96­
hour
EC50(
growth
rate)
=
>
3.2
96­
hour
NOEC(
growth
rate)
=
>
3.2
Good
Test
substance
was
PFOS
potassium
salt.

Measured
exposure
concentrations.
39
ENV/
JM/
RD(
2002)
17/
FINAL
72
Table
12.
Sub­
chronic/
chronic
(
prolonged/
long­
term)
toxicity
data
for
PFOS
determined
in
tests
with
freshwater
algae
Species
Protocol
Results
(
mg/
l)
Study
Standard
Comments
Study
Ref.

No.

Selenastrum
capricornutum
OECD
201,
US
EPA
600/
9­
78­

018
&
ASTM­
E­
35.23
(
Static)
14­
day
EC50
cell
density
=
95
14­
day
NOECcell
density
=
<
26
14­
day
EC10
cell
density
=
16
Acceptable
Test
substance
was
PFOS
potassium
salt.

Result
not
expressed
relative
to
growth
rate.

No
analysis
of
exposure
but
result
agrees
with
those
obtained
in
studies
2
and
24.
13
Table
13.
Acute
(
short­
term
exposure)
toxicity
data
for
PFOS
determined
in
tests
with
higher
plants
Medium
Species
Protocol
Result
(
mg/
l)
Study
Standard
Comments
Study
Ref.

No.

Freshwater
Lemna
gibba
G3
(
Duckweed)
OPPTS
850.4400
7­
day
IC50
=
108
Good
Test
substance
was
PFOS
potassium
salt.

Measured
exposure
concentrations.
37
ENV/
JM/
RD(
2002)
17/
FINAL
73
4.2
Effects
on
Other
Aquatic
Organisms
4.2.1
Amphibians
Data
summarised
in
Table
14
were
obtained
in
an
embryo
teratogenesis
assay
carried
out
on
PFOS
potassium
salt
with
Xenopus
laevis
(
African
clawed
frog).
Exposure
of
the
embryos
for
96
hours
resulted
in
an
LC50
for
mortality
of
13.8
mg/
l
and
an
EC50
for
malformations
of
12.1
mg/
l.
The
minimum
concentration
that
inhibited
growth
was
7.97
mg/
l.
and
the
teratogenic
index
was
calculated,
as
the
ratio
of
the
96­
hour
LC50
to
the
96­
hour
EC50,
to
be
1.1.
The
latter
value
indicates
that
PFOS
has
a
low
potential
to
be
a
developmental
hazard
in
this
species.
ENV/
JM/
RD(
2002)
17/
FINAL
74
Table
14.
Toxicity
data
for
PFOS
determined
in
an
embryo
teratogenesis
assay
with
the
amphibian,








	





Medium
Species
Protocol
Result
(
mg/
l)
Study
Standard
Comments
Study
Ref.

No.

Freshwater
Xenopus
laevis
(
African
clawed
frog)
ASTM
E1439­
91
96­
hour
LC50
=
13.8
96­
hour
EC50(
malformations)
=
12.1
Minimum
concentration
to
inhibit
growth
=
7.97
Teratogenic
index
1.1
Acceptable
Test
substance
was
PFOS
potassium
salt.

Measured
exposure
concentrations.

In­
life
phases
were
not
subject
to
GLP.
40
ENV/
JM/
RD(
2002)
17/
FINAL
75
4.2.2
Sediment
Dwelling
Invertebrates
No
data
are
available
for
effects
on
sediment
dwelling
invertebrates.
It
is
possible
that
a
predicted
no
effect
concentration
(
PNEC)
for
sediment­
dwelling
invertebrates
could
be
determined
by
applying
equilibrium
partitioning
models
to
the
data
for
water
column
organisms.
However,
as
discussed
in
Annex
3,
this
is
unlikely
to
be
feasible
at
the
present
time.

4.2.3
Bacteria
Data
summarised
in
Table
15
were
obtained
from
2
reports
describing
Microtox
 
studies
(
study
report
reference
numbers
18
and
22).
The
Microtox
 
procedure
assesses
light
output
inhibition
from
the
luminescent
marine
bacterium
Photobacterium
phosphoreum,
following
exposure
to
a
toxicant
over
a
short
exposure
period
(
30
minutes
in
the
case
of
these
two
studies).
Both
studies
are
of
an
unacceptable
standard
for
assessing
the
hazard
of
PFOS
(
lithium
salt)
to
bacteria
because
of
uncertainty
over
the
true
exposure
concentrations
and
in
one
case
because
of
the
presence
of
diethylene
glycol
in
the
test
sample.
There
are
also
more
general
concerns
over
the
relevance
of
the
test
system
and
test
species
for
determining
effects
for
this
particular
group
of
organisms.
No
toxic
effects
were
observed
in
either
test
at
nominal
concentrations
(>
250
mg/
l)
that
were
significantly
in
excess
of
the
solubility
of
PFOS
salts
in
saline
medium
(
2.5
to
20
mg/
l,
depending
on
purity/
salinity).
ENV/
JM/
RD(
2002)
17/
FINAL
76
Table
15.
Toxicity
data
for
PFOS
determined
in
Microtox
tests
Species
Protocol
Results
(
mg/
l)
Study
Standard
Comments
Study
Ref.

No.

Photobacterium
phosphoreum
(
Microtox
 
)
Microbics
Microtox
 
"
BASIC"
15­
minute
EC50
=
>
250
30­
minute
EC50
=
>
250
Unacceptable
Test
substance
was
PFOS
lithium
salt.

Nominal
exposure
concentrations.

Initial
stock
solution
prepared
at
concentration
that
was
likely
to
exceed
solubility
in
saline
water
(
2.5
to
20
mg/
l,

depending
on
purity/
salinity)
It
is
therefore
possible
that
actual
exposure
concentrations
were
much
lower
than
nominal.

Result
indicates
probable
absence
of
toxicity
at
limit
of
solubility.
18
30­
minute
EC50
=
>
280
Unacceptable
Test
substance
was
PFOS
potassium
salt.

Nominal
exposure
concentrations.

Data
questionable
in
view
of
use
of
diethylene
glycol
to
aid
dosing.

Test
concentrations
likely
to
be
in
excess
of
solubility
in
saline
medium
(
2.5
to
20
mg/
l,
depending
on
purity/
salinity).

Result
is
consistent
with
that
obtained
in
study
18
indicating
probable
absence
of
toxicity
at
limit
of
solubility.
22
ENV/
JM/
RD(
2002)
17/
FINAL
77
4.2.4
Activated
Sludge
Microorganisms
Data
summarised
in
Table
16
were
obtained
from
3
reports
describing
activated
sludge
respiration
inhibition
studies
(
study
report
reference
numbers
6,
19
and
21).
Only
study
reference
number
9
was
considered
to
be
of
an
acceptable
standard
for
assessing
the
hazard
of
PFOS
to
activated
sludge.
Studies
19
and
21
had
significant
inadequacies
in
test
procedures
related
to
test
conditions,
duration
of
the
exposure
and/
or
absence
of
a
reference
substance.

The
3­
hour
IC50
value
for
PFOS
(
potassium
salt)
determined
in
the
acceptable
study
was
>
905
mg/
l
(
nominal
concentration).
ENV/
JM/
RD(
2002)
17/
FINAL
78
Table
16.
Toxicity
data
for
PFOS
determined
in
Activated
Sludge
Respiration
Inhibition
tests
Test
system
Protocol
Results
(
mg/
l)
Study
Standard
Comments
Study
Ref.
No.

Activated
Sludge
OECD
209
3­
hour
IC50
=
>
905
(
Nom.)
Good
Test
substance
was
PFOS
potassium
salt.

Nominal
exposure
concentrations
of
0.9
 
905
mg/
l
tested.
Highest
test
concentration
gave
the
maximum
inhibition
of
38.8%.
6
OECD
209
3­
hour
IC50
=
>
245
(
as
a
nom.
24.5%
solution
in
water)
Unacceptable
Test
substance
was
PFOS
potassium
salt.

Nominal
exposure
concentrations.

Test
temperature
not
controlled
and
fell
well
below
requirements
of
standard
guidelines.

Respiration
rates
extrapolated
to
20oC.

Oxygenation
was
inadequate
during
the
first
30
minutes
of
the
3­
hour
test.

No
inhibitory
effect
observed.
19
Not
noted
7­
minute
IC50
=
>
250
(
as
a
nom.
25%
solution
in
water)
Unacceptable
Test
substance
was
PFOS
DEA
salt.

Nominal
exposure
concentrations.

Exposure
period
inadequate
by
current
standards.

No
reference
substance
used.

No
inhibitory
effect
observed.
21
ENV/
JM/
RD(
2002)
17/
FINAL
79
4.3
Effects
on
Terrestrial
Organisms
4.3.1
Soil­
dwelling
Invertebrates
No
data
are
available
for
effects
on
soil
dwelling
invertebrates.
It
is
however
possible
that
a
predicted
no
effect
concentration
(
PNEC)
for
soil­
dwelling
invertebrates
could
be
determined
by
applying
equilibrium
partitioning
models
to
the
data
for
water
column
organisms.
However,
as
discussed
in
Annex
3,
this
is
unlikely
to
be
feasible
at
the
present
time.
It
is
noted
that
an
earthworm
acute
toxicity
and
uptake
study
is
in
the
planning
stages,
and
a
final
report
should
be
available
in
2002.

4.3.2
Terrestrial
Plants
No
data
are
available
for
effects
on
terrestrial
plants.
It
is
however
possible
that
a
predicted
no
effect
concentration
(
PNEC)
for
terrestrial
plants
could
be
determined
by
applying
equilibrium
partitioning
models
to
the
data
for
aquatic
plants.
However,
as
discussed
in
Annex
3,
this
is
unlikely
to
be
feasible
at
the
present
time.
It
is
noted
that
a
terrestrial
plant
toxicity
and
uptake
study
should
be
available
in
2002.

4.3.3
Birds
Data
summarised
in
Table
17
were
from
2
reports
describing
dietary
acute
studies
with
the
Mallard
duck,
Anas
platyrhynchos,
and
the
Northern
Bobwhite
quail,
Colinus
virginianus
(
study
report
reference
numbers
11
and
12).
Birds
in
both
tests
were
exposed
to
PFOS
potassium
salt
in
their
diets
for
5
days
and
observations
were
made
after
3
or
17
days.
Both
studies
are
considered
to
be
of
an
acceptable
standard
for
assessing
the
acute
toxic
hazard
of
PFOS
to
birds
via
dietary
exposure.

The
lowest
acute
dietary
LC50
value
of
220
mg/
kg
of
food
was
determined
in
the
test
with
the
quail.
This
value
was
approximately
half
that
obtained
in
the
test
with
the
duck.
The
lowest
NOEC
of
37
mg/
kg
of
food
for
effects
on
body
weight
was,
in
contrast,
obtained
in
the
test
with
the
duck.
Once
again
this
value
was
approximately
half
that
obtained
with
the
other
species
 
the
quail.

It
is
noted
that
avian
reproductive
data
will
be
submitted
for
review
in
2002.
ENV/
JM/
RD(
2002)
17/
FINAL
80
Table
17.
Acute
dietary
toxicity
data
for
PFOS
determined
in
tests
with
birds
Test
species
Protocol
Results
(
mg/
kg
of
food)
Study
Standard
Comments
Study
Ref.
No.

Anas
platyrhynchos
(
Mallard
duck)
OECD
205,
OPPTS
850.2200
&
FIFRA
E
71­
2
LC50
=
628
NOECmortality
=
146
NOECbody
weight
=
37
Good
Test
substance
was
PFOS
potassium
salt.

Measured
exposure
concentration
in
food.

5
days
exposure
followed
by
3
or
17
days
observation
11
Colinus
virginianus
(
Northern
Bobwhite
quail)
OECD
205,
OPPTS
850.2200
&
FIFRA
E
71­
2
LC50
=
220
NOECmortality
=
73
NOECbody
weight
=
73
Good
Test
substance
was
PFOS
potassium
salt.

Measured
exposure
concentration
in
food.

5
days
exposure
followed
by
3
or
17
days
observation
12
ENV/
JM/
RD(
2002)
17/
FINAL
81
4.3.4
Bees
Data
summarised
in
Table
18
were
obtained
in
acute
oral
and
contact
toxicity
tests
carried
out
with
the
Honey
bee
(
Apis
mellifera)
on
PFOS
potassium
salt.
The
studies
fully
meet
the
required
standard
for
acceptability
of
the
data.

The
acute
oral
test
yielded
a
72­
hour
LD50
for
ingestion
of
PFOS
of
0.40
µ
g/
bee
and
a
72­
hour
NOEL
of
0.21
µ
g/
bee.
The
contact
test
yielded
a
96­
hour
LD50
of
4.78
µ
g/
bee
and
a
96­
hour
NOEL
of
1.93
µ
g/
bee.
The
respective
LD50
values
indicate
moderate
and
high
orders
of
toxicity
of
PFOS
to
bees
when
administered
via
these
routes.
ENV/
JM/
RD(
2002)
17/
FINAL
82
Table
18.
Acute
oral
and
contact
toxicity
data
for
PFOS
determined
in
tests
with
Honey
bees
Test
species
Protocol
Results
(
µ
g/
bee)
Study
Standard
Comments
Study
Ref.
No.

Apis
mellifera
(
Honey
bee)
OECD
213,

EPPO
170
(
Oral)
72­
hour
LD50
=
0.40
72­
hour
NOEL
=
0.21
Good
Test
substance
was
PFOS
potassium
salt.

Nominal
exposure
following
feeding
on
50%
w/
v
sucrose.
34
Apis
mellifera
(
Honey
bee)
OECD
214,

EPPO
170,

OPPTS
850.3020
(
draft)
(
Contact)
96­
hour
LD50
=
4.78
96­
hour
NOEL
=
1.93
Good
Test
substance
was
PFOS
potassium
salt.

Nominal
exposure
following
direct
application
of
test
substance
dissolved
in
acetone
to
thorax.
35
ENV/
JM/
RD(
2002)
17/
FINAL
83
5.0
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Final
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May
18,
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Olsen,
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ENV/
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6295.7;
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ENV/
JM/
RD(
2002)
17/
FINAL
89
Annex
1.
Ecological
Studies
The
following
lists
of
study
reports
were
contained
on
a
3M
Fluorochemical
EPA
Submissions
CDs
dated
12th
July
2000,
28th
June
2001,
and
February
2002
12th
July
2000
1.
96­
Hour
Static
Acute
Toxicity
Test
with
the
Fathead
Minnow
(
Pimephales
promelas)
2.
96­
Hour
Toxicity
Test
with
the
Freshwater
Alga
(
Selenastrum
capricornutum)
3.
48­
Hour
Static
Acute
Toxicity
Test
with
the
Cladoceran
(
Daphnia
magna)
4.
96­
Hour
Shell
Deposition
Test
with
the
Eastern
Oyster
(
Crassostrea
virginica)
5.
96­
Hour
Static
Acute
Toxicity
Test
with
the
Freshwater
Mussel
(
Unio
complamatus)
6.
Activated
Sludge,
Respiration
Inhibition
Test
7.
96­
Hour
Static
Acute
Toxicity
Test
with
the
Saltwater
Mysid
(
Mysidopsis
bahia)
8.
Early
Life­
Stage
Toxicity
Test
with
the
Fathead
Minnow
(
Pimephales
promelas)
9.
Semi­
Static
Life­
Cycle
Toxicity
Test
with
the
Cladoceran
(
Daphnia
magna)
10.
Flow­
through
Life­
Cycle
Toxicity
Test
with
the
Saltwater
Mysid
(
Mysidopsis
bahia)
11.
Dietary
LC50
Study
with
the
Mallard
12.
Dietary
LC50
Study
with
the
Northern
Bobwhite
13.
Multi­
Phase
Exposure
/
Recovery
Algal
Assay
Test
14.
The
Effects
of
Continuous
Aqueous
Exposure
to
14C­
78.02
on
Hatchability
of
Eggs
and
Growth
and
Survival
of
Fry
of
Fathead
Minnow
(
Pimephales
promelas
/
Summary
of
histopathological
examinations
of
Fathead
Minnow
(
Pimephales
promelas)
exposed
to
78.02
for
30
Days
15.
Effect
of
Potassium
Perfluorooctanesulfonate
on
Survival,
etc.
(
Daphnid
reproduction)
16.
Pimephales
promelas
96­
hour
Toxicity
Test
Data
Summary.
Sample
FC­
94­
X
(
Li
salt
of
PFOS)
17.
48­
HR
Acute
Toxicity
to
Daphnia,
Daphnia
magna.
FC­
94­
X
(
Li
salt
of
PFOS)
18.
Microbics
Microtox
Toxicity
Test.
Sample
:
FC­
94­
X
(
Li
salt
of
PFOS)
19.
Evaluation
of
FC­
94­
X
by
OECD
Activated
Sludge
Respiration
Inhibition
Test
#
209
Review
of
OECD
209
and
BOD/
COD
Test
Results
for
FC­
94­
X,
test
data
sheets
(
Li
salt
of
PFOS)
20.
96­
Hour
Acute
Toxicity
Test
on
Bluegill
Sunfish
(
FC­
99,
DEA
salt
of
PFOS)
21.
Acute
Toxicity
to
Activated
Sludge
(
FC­
99,
DEA
salt
of
PFOS)
22.
Microtox
data
for
FM­
3820
(
28%
PFOS)
23.
Acute
Toxicity
to
Daphnia
magna
for
FM­
3820
(
28%
PFOS)
24.
Toxicity
to
Algae
(
Selenastrum
capricornutum)
for
FC­
3820
(
28%
PFOS)
25.
Final
Comprehensive
Report
:
FC­
95
26.
Data
from
Fathead
Minnow
Study
on
FC­
93
(
25%
NH4
salt
of
PFOS
in
IPA
and
water),
3M
Environmental
Lab,
Aug.
2,
1974.
27.
Data
from
Fathead
Minnow
Study
on
FC­
93
(
25%
NH4
salt
of
PFOS
in
IPA
and
water),
3M
Environmental
Lab,
Oct.
19,
1974.
28.
Acute
toxicity
of
P3025
developmental
material
to
Fathead
minnow
(
Pimephales
promelas).
29.
Acute
toxicity
of
P3025
developmental
material
to
Daphnia
magna.

28th
June
2001
30.
Acute
toxicity
of
PFOS
to
Rainbow
trout
in
saltwater
31.
Acute
toxicity
of
PFOS
to
Rainbow
trout
in
freshwater
32.
Acute
toxicity
of
PFOS
to
Artemia
sp.
33.
Acute
toxicity
of
PFOS
to
Daphnia
magna
34.
Perfluorooctanesulfonate,
Potassium
salt
(
PFOS):
An
acute
oral
toxicity
study
with
the
Honey
bee
35.
Perfluorooctanesulfonate,
Potassium
salt
(
PFOS):
An
acute
contact
toxicity
study
with
the
Honey
bee
36.
PFOS:
A
96­
hour
toxicity
test
with
the
freshwater
alga
(
Anabaena
flos­
aquae)
37.
PFOS:
A
7­
day
toxicity
test
with
Duckweed
(
Lemna
gibba
G3)
38.
PFOS:
A
96­
hour
toxicity
test
with
freshwater
diatom
(
Navicula
pelliculosa)
ENV/
JM/
RD(
2002)
17/
FINAL
90
39.
PFOS:
A
96­
hour
toxicity
test
with
the
marine
diatom
(
Skeletonema
costatum)
40.
PFOS:
A
frog
embryo
teratogenesis
assay
 
Xenopus
(
FETAX)
41.
Perfluorooctanesulfonate,
Potassium
salt
(
PFOS):
A
flow­
through
bioconcentration
test
with
the
Bluegill
(
Lepomis
macrochirus)

February
2002
42.
Perfluorooctanesulfonate,
Potassium
salt
(
PFOS):
96­
Hour
Static
Acute
Toxicity
Test
with
the
Rainbow
Trout
(
Oncorhynchus
mykiss)
in
freshwater
43.
Perfluorooctanesulfonate,
Potassium
salt
(
PFOS):
96­
Hour
Semi­
Static
Acute
Toxicity
Test
with
the
Sheepshead
Minnow
(
Cyprinodon
variegatus)
in
saltwater
ENV/
JM/
RD(
2002)
17/
FINAL
91
Annex
2.
Robust
Summaries
of
Key
Ecotoxicology
Studies
Study
reference
number
Title
1
96­
Hour
Static
Acute
Toxicity
Test
with
the
Fathead
Minnow
(
Pimephales
promelas)
2
96­
Hour
Toxicity
Test
with
the
Freshwater
Alga
(
Selenastrum
capricornutum)
3
48­
Hour
Static
Acute
Toxicity
Test
with
the
Cladoceran
(
Daphnia
magna)
4
96­
Hour
Shell
Deposition
Test
with
the
Eastern
Oyster
(
Crassostrea
virginica)
5
96­
Hour
Static
Acute
Toxicity
Test
with
the
Freshwater
Mussel
(
Unio
complamatus)
6
Activated
Sludge,
Respiration
Inhibition
Test
7
96­
Hour
Static
Acute
Toxicity
Test
with
the
Saltwater
Mysid
(
Mysidopsis
bahia)
8
Early
Life­
Stage
Toxicity
Test
with
the
Fathead
Minnow
(
Pimephales
promelas)
9
Semi­
Static
Life­
Cycle
Toxicity
Test
with
the
Cladoceran
(
Daphnia
magna)
10
Flow­
through
Life­
Cycle
Toxicity
Test
with
the
Saltwater
Mysid
(
Mysidopsis
bahia)
11
Dietary
LC50
Study
with
the
Mallard
12
Dietary
LC50
Study
with
the
Northern
Bobwhite
13
Multi­
Phase
Exposure
/
Recovery
Algal
Assay
Test
14
The
Effects
of
Continuous
Aqueous
Exposure
to
14C­
78.02
on
Hatchability
of
Eggs
and
Growth
and
Survival
of
Fry
of
Fathead
Minnow
(
Pimephales
promelas
/
Summary
of
histopathological
examinations
of
Fathead
Minnow
(
Pimephales
promelas)
exposed
to
78.02
for
30
Days
15
Effect
of
Potassium
Perfluorooctanesulfonate
on
Survival,
etc.
(
Daphnid
reproduction)
16
Pimephales
promelas
96­
hour
Toxicity
Test
Data
Summary.
Sample
FC­
94­
X
(
Li
salt
of
PFOS)
17
48­
HR
Acute
Toxicity
to
Daphnia,
Daphnia
magna.
FC­
94­
X
(
Li
salt
of
PFOS)
20
96­
Hour
Acute
Toxicity
Test
on
Bluegill
Sunfish
(
FC­
99,
DEA
salt
of
PFOS)
23
Acute
Toxicity
to
Daphnia
magna
for
FM­
3820
(
28%
PFOS)
26
Data
from
Fathead
Minnow
Study
on
FC­
93
(
25%
NH4
salt
of
PFOS
in
IPA
and
water),
3M
Environmental
Lab,
Aug.
2,
1974.
27
Data
from
Fathead
Minnow
Study
on
FC­
93
(
25%
NH4
salt
of
PFOS
in
IPA
and
water),
3M
Environmental
Lab,
Oct.
19,
1974.
28
Acute
toxicity
of
P3025
developmental
material
to
Fathead
minnow
(
Pimephales
promelas).
29
Acute
toxicity
of
P3025
developmental
material
to
Daphnia
magna.
30
Acute
toxicity
of
PFOS
to
Rainbow
trout
in
saltwater
31
Acute
toxicity
of
PFOS
to
Rainbow
trout
in
freshwater
32
Acute
toxicity
of
PFOS
to
Artemia
sp.
33
Acute
toxicity
of
PFOS
to
Daphnia
magna
34
Perfluorooctanesulfonate,
Potassium
salt
(
PFOS):
An
acute
oral
toxicity
study
with
the
Honey
bee
35
Perfluorooctanesulfonate,
Potassium
salt
(
PFOS):
An
acute
contact
toxicity
study
with
the
Honey
bee
36
PFOS:
A
96­
hour
toxicity
test
with
the
freshwater
alga
(
Anabaena
flos­
aquae)
37
PFOS:
A
7­
day
toxicity
test
with
Duckweed
(
Lemna
gibba
G3)
38
PFOS:
A
96­
hour
toxicity
test
with
freshwater
diatom
(
Navicula
pelliculosa)
39
PFOS:
A
96­
hour
toxicity
test
with
the
marine
diatom
(
Skeletonema
costatum)
40
PFOS:
A
frog
embryo
teratogenesis
assay
 
Xenopus
(
FETAX)
41
Perfluorooctanesulfonate,
Potassium
salt
(
PFOS):
A
flow­
through
bioconcentration
test
with
the
Bluegill
(
Lepomis
macrochirus)
42
Perfluorooctanesulfonate,
Potassium
salt
(
PFOS):
96­
Hour
Static
Acute
Toxicity
Test
with
the
Rainbow
Trout
(
Oncorhynchus
mykiss)
in
freshwater
43
Perfluorooctanesulfonate,
Potassium
salt
(
PFOS):
96­
Hour
Semi­
Static
Acute
Toxicity
Test
with
the
Sheepshead
Minnow
(
Cyprinodon
variegatus)
in
saltwater
44
Bioconcentration
test
of
Salt
(
Na,
K,
Li)
of
perfluoroalkyl
(
C=
4­
12)
sulfonic
acid
[
This
test
was
performed
using
Perfluorooctane
sulfonic
acid,
potassium
salt
(
Test
substance
number
K­
1520)]
in
carp
ENV/
JM/
RD(
2002)
17/
FINAL
92
Robust
Study
Report
Reference
No.
1
 
96­
Hour
Static
Acute
Toxicity
Test
with
the
Fathead
Minnow
(







	








	


)

TEST
SUBSTANCE
Identity:
Potassium
perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
The
test
substance
is
a
white
powder.
Sample
was
taken
from
3M
lot
number
217.
Sample
was
stored
under
ambient
conditions
prior
to
testing.
Purity
determined
to
be
90.49%
by
LC/
MS,
1H­
HMR,
19FNMR
and
elemental
analyses
techniques.

METHOD
Method:
OECD
203
and
OPPTS
850.1075
Type:
Static
acute
GLP:
Yes
Year
completed:
Study
completed
1999.
Report
completed
2000
Species:
Pimephales
promelas
Supplier:
In­
house
cultures,
Wildlife
International,
Ltd.,
Easton,
MD,
USA
Analytical
monitoring:
PFOS
measured
at
0,
48,
96­
hours
Exposure
period:
96­
hours
Statistical
methods:
LC50
values
calculated,
when
possible,
by
probit
analysis,
moving
average
method
or
binomial
probability
with
non­
linear
interpolation
using
the
computer
software
of
C.
E.
Stephan.
Test
fish
age:
Approximately
126
days
old
Length
and
weight:
35
(
30­
38)
mm,
0.36
(
0.21­
0.49)
g
Loading:
0.24
g
fish/
L
Pretreatment:
None
Test
Conditions
Dilution
water:
0.45
µ
m
filtered
well
water
Dilution
water
chemistry
(
during
the
4­
week
period
immediately
preceding
the
test):
Hardness:
131
(
128­
136)
mg/
L
as
CaCO3
Alkalinity:
177
(
176­
178)
mg/
L
as
CaCO3
pH:
8.3
TOC:
<
1.0
mg/
L
Conductivity:
311
(
310­
315)
µ
mhos/
cm
Stock
and
test
solution
preparation:
Primary
stock
prepared
in
dilution
water
at
27
mg/
L
and
mixed
for
~
22
hours
prior
to
use.
After
mixing,
primary
stock
solution
was
proportionally
diluted
with
dilution
water
to
prepare
the
four
additional
test
concentrations.
Concentrations
dosing
rate:
Once
Stability
of
the
test
chemical
solutions:
Extremely
stable
Exposure
vessels:
25L
polyethylene
aquaria
containing
approximately
15L
of
test
solution;
water
depth
approximately
17.6
cm.
Number
of
replicates:
two
Number
of
fish
per
replicate:
ten
Number
of
concentrations:
five
plus
a
negative
control
Water
chemistry
during
the
study:
Dissolved
oxygen
range
(
0
 
96
hours):
7.8
 
8.8
mg/
L
(
control
exposure)
ENV/
JM/
RD(
2002)
17/
FINAL
93
7.7
 
9.0
mg/
L
(
28
mg/
L
exposure)
pH
range
(
0
 
96
hours)
8.3
 
8.6
(
control
exposure)
8.4
 
8.5
(
28
mg/
L
exposure)
Test
temperature
range
(
0
 
96
hours)
20.4
 
22.1oC
(
control
exposure)
21.3
 
22.3
oC
(
28
mg/
L
exposure)
Method
of
calculating
mean
measured
concentrations:
arithmetic
mean
RESULTS
Nominal
concentrations:
Negative
control,
3.6,
5.9,
9.9,
16,
27
mg/
L
Measured
concentrations:
<
LOQ,
3.3,
5.6,
9.5,
17,
28
mg/
L
Element
value:
(
95%
confidence
interval
is
given
in
brackets)
24­
hour
LC50
=
>
28
mg/
L
(
C.
I.
not
calculable)
48­
hour
LC50
=
>
28
mg/
L
(
C.
I.
not
calculable)
72­
hour
LC50
=
27
(
22
 
41)
mg/
L
96­
hour
LC50
=
9.5
(
8.0
 
11)
mg/
L
All
element
values
based
on
mean
measured
concentrations
Statistical
evaluation
of
mortality:
Confidence
limits
for
24
and
48­
hours
could
not
be
calculated
due
to
lack
of
mortality.
The
72­
hour
LC50
value
is
questionable
because
a
concentration­
effect
relationship
was
not
demonstrated
over
a
reasonable
range
of
percent
dead.
The
24
and
48­
hour
LC50
values
were
determined
by
visual
interpretation.
Probit
was
used
to
calculate
the
72­
hour
LC50
and
Moving
Average
for
the
96­
hour
LC50.

Analytical
methodology:
Analyses
of
test
solutions
were
performed
at
Wildlife
International
Ltd.
using
high
performance
liquid
chromatography
with
mass
spectrometric
detection
(
HPLC/
MS).
When
determining
the
concentration
of
the
test
substance
in
the
test
solutions,
the
same
and
most
prominent
peak
response
for
perfluorooctanesulfonate
was
used.
No
attempt
was
made
to
quantify
on
the
basis
of
individual
isomeric
components.
The
LOQ
(
limit
of
quantitation)
was
0.458
mg/
L
in
this
study.
The
mean
percent
recovery
of
matrix
fortifications
analyzed
concurrently
during
sample
analysis
was
97.9.
Samples
collected
at
test
initiation
had
measured
values
from
85.3
to
117%
of
nominal.
Measured
values
for
samples
taken
at
48
hours
ranged
from
86.3
to
101%
of
nominal.
Measured
values
for
samples
taken
at
96
hours
ranged
from
87.6
to
98.3%
of
nominal.

Summary
of
analytical
chemistry
data:

Nominal
Test
Concentration,
mg/
L
Measured
Duplicate
Values
at
0,
48,
and
96­
hours,
Respectively,
mg/
L
Mean
Measured
Concentration
mg/
L
Percent
of
Nominal
Negative
Control
All
<
LOQ
<
LOQ
­
3.6
3.16,
3.53,
3.08,
3.22,
3.46,
3.13
3.3
92
5.9
6.05,
5.07,
5.48,
5.89,
5.70,
5.55
5.6
95
9.9
8.99,
9.47,
9.88,
9.33,
9.70,
9.52
9.5
96
16
18.2,
19.3,
15.0,
15.6,
14.8,
16.2
17
106
27
28.5,
28.5,
27.0,
27.8,
26.8,
26.6
28
104
Biological
observations
after
96­
hours:
Fish
in
the
negative
control
and
the
3.3
mg/
L
exposure
concentration
appeared
normal.
Some
or
all
of
the
surviving
fish
were
observed
to
be
swimming
erratically
(
4/
16
in
5.6
mg/
L
exposure,
10/
10
in
9.5
mg/
L,
4/
4
in
17
mg/
L)
at
test
termination.
ENV/
JM/
RD(
2002)
17/
FINAL
94
Cumulative
percent
mortality:

Mean
Measured
Test
Concentration
mg/
L
24­
hours
48­
hours
72­
hours
96­
hours
Neg.
Control
0
0
0
0
3.3
0
0
0
0
5.6
0
0
0
20
9.5
0
0
0
50
17
0
0
15
80
28
0
0
50
100
Lowest
concentration
causing
100%
mortality:
28
mg/
L
Mortality
of
controls:
None
CONCLUSIONS
The
potassium
perfluorooctanesulfonate
96­
hour
LC50
for
fathead
minnow
was
determined
to
be
9.5
mg/
L
with
a
95%
confidence
interval
of
8.0
 
11
mg/
L.
The
96­
hour
no
mortality
and
no
effects
concentration
was
3.3
mg/
L.

Submitter:
3M
Company,
Environmental
Laboratory
P.
O.
Box
33331
St.
Paul,
MN
55133,
USA
DATA
QUALITY
Reliability:
Klimisch
ranking
=
1
REFERENCES
This
study
was
conducted
at
Wildlife
International
Ltd.,
Easton,
MD
at
the
request
of
the
3M
Company.

OTHER
Last
changed:
5/
3/
00
ENV/
JM/
RD(
2002)
17/
FINAL
95
Robust
Study
Report
Reference
No.
2
 
96­
Hour
Toxicity
Test
with
the
Freshwater
Alga
(


	






















)

TEST
SUBSTANCE
Identity:
Potassium
perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonicacid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
The
test
substance
is
a
white
powder.
Sample
was
taken
from
3M
lot
number
217.
Sample
was
stored
under
ambient
conditions
prior
to
testing.
Purity
determined
to
be
90.49%
by
LC/
MS,
'
H­
HMR,
19F­
NMR
and
elemental
analyses
techniques.

METHOD
Method:
OECD
201,
OPPTS
850.5400,
ASTM
1218­
90E
Test:
Static
acute
GLP:
Yes
Year
completed:
Study
completed
1999.
Report
completed
2000
Species:
Selenastrum
capricornutum
Source:
Originally
from
The
Culture
Collection
of
Algae
at
the
University
of
Texas
at
Austin,
maintained
in
culture
medium
at
Wildlife
International
Ltd.,
Easton,
MD,
USA
Analytical
monitoring:
PFOS
measured
at
0,
72,
96­
hours
Element
basis:
Reported
three
ways:
number
of
cells/
ml,
area
under
the
growth
curve
and
growth
rate
Exposure
period:
96­
hours
Start
date:
4/
12/
99
End
date:
4/
16/
99
Analytical
monitoring:
Test
concentrations
measured
at
0,
72
and
96­
hours.
Test
organisms
laboratory
culture:
Algae
cultures
had
been
actively
growing
in
freshwater
algal
culture
medium
for
at
least
two
weeks
prior
to
test
initiation.
Stock
nutrient
solutions
were
prepared
by
adding
reagent­
grade
chemicals
to
reverse
osmosis­
purified
well
water.
Test
Conditions:
Test
temperature
range:
23.6­
25.8oC
Growth
medium:
ASTM
Standard
Guide1218­
90E,
1990
Compound
Nominal
concentration
Units
MgCl26H20
12.16
mg/
l
CaCl22H2O
4.40
mg/
l
H3BO3
0.1856
mg/
l
MnCl24H2O
0.416
mg/
l
ZnCl2
3.28
µ
g/
l
FeCl26H2O
0.1598
mg/
l
CoCl26H2O)
1.428
µ
g/
l
Na2MoO42H2O
7.26
µ
g/
l
CuCl22H2O
0.012
µ
g/
l
Na2EDTA2H2O
0.300
mg/
l
NaNO3
25.50
mg/
l
MgSO47H2O
14.70
mg/
l
K2HPO4
1.044
mg/
l
ENV/
JM/
RD(
2002)
17/
FINAL
96
NaHCO3
15.0
mg/
l
Dilution
water
source:
Wildlife
International
Ltd.
well
water
purified
by
reverse
osmosis.
The
test
medium
was
prepared
by
adding
the
appropriate
volumes
of
stock
nutrient
solutions
to
purified
well
water.
The
pH
of
the
medium
was
adjusted
to
7.5+
0.1
using
10%
HCI
and
the
medium
was
sterilized
by
filtration
(
0.22
µ
m)
prior
to
use.
Stock
and
test
solution
preparation:
A
primary
stock
solution
was
prepared
in
algal
medium
at
a
concentration
of
183
mg/
L.
The
primary
stock
solution
was
stirred
with
a
magnetic
stir
plate
for
approximately
24
hours.
After
mixing,
the
primary
stock
solution
was
proportionally
diluted
with
algal
medium
to
prepare
the
five
additional
test
concentrations.
All
final
test
solutions
appeared
clear
and
colorless.
Exposure
vessels:
Sterile
250
mL
polycarbonate
Erlenmeyer
flasks
plugged
with
foam
stoppers
containing
100
mL
of
test
solution.
Agitation:
Shaken
continuously
at
100
rpm
Number
of
replicates:
three
Initial
algal
cell
loading:
1.0
X
104
cells/
mL
Number
of
concentrations:
six
plus
a
negative
control
plus
an
abiotic
control
at
the
highest
concentration
tested
Water
chemistry:
pH
range
(
0­
96
hours)
7.5
­
8.1
(
control
exposure)
7.4
­
7.5
(
179
mg/
L
exposure)
Test
temperature
range
(
0­
96
hours)
23.6
­
25.8oC
Light
levels
(
0­
96
hours)
3870
­
4610
lux
from
continuous
cool­
white
fluorescent
lighting
Method
of
calculating
mean
measured
concentrations:
arithmetic
mean
RESULTS
Nominal
concentrations:
Negative
control,
5.7,
11,
23,
46,
91,183
mg/
L
plus
183
mg/
L
abiotic
control.
Measured
concentrations:
<
LOQ,
5.5,
11,
21,
44,
86,
179,
169
mg/
L
Element
value:
(
95%
confidence
interval
is
given
in
brackets)
24­
hour
EC50
(
cell
density)
=
163
(
74
­
191)
mg/
L
24­
hour
EbC50
(
area
under
curve)
=
122
(
19
­
176)
mg/
L
24­
hour
EC50
(
growth
rate)
=
136
(
30
­
204)
mg/
L
48­
hour
EC50
(
cell
density)
=
81
(
72
­
90)
mg/
L
48­
hour
EbC50
(
area
under
curve)
=
84
(
67
­
146)
mg/
L
48­
hour
ErC50
(
growth
rate)
=
142
(
107
­
185)
mg/
L
72­
hourEC10
(
cell
density)
=
37
(<
O
­
64)
mg/
L
72­
hourEbC10
(
area
under
curve)
=
46
(<
O
­
56)
mg/
L
72­
hour
ErC10
(
growth
rate)
=
53
(
23
­
64)
mg/
L
72­
hourEC50
(
cell
density)
=
70
(
44
­
78)
mg/
L
72­
hourEbC50
(
area
under
curve)
=
74
(
55
­
82)
mg/
L
72­
hour
ErC50
(
growth
rate)
=
120
(
103
­
132)
mg/
L
72­
hourEC90
(
cell
density)
=
153
(
130
­
165)
mg/
L
72­
hourEbC90
(
area
under
curve)
=
165
(
145
­
176)
mg/
L
72­
hour
ErC90
(
growth
rate)
=
>
179
mg/
L
(
C.
I.
not
calculable)
96­
hourEC10
(
cell
density)
=
49
(
43
­
50)
mg/
L
96­
hourEBC10
(
area
under
curve)
=
49
(
40
­
50)
mg/
L
96­
hourErC10
(
growth
rate)
=
59
(
54
­
63)
mg/
L
ENV/
JM/
RD(
2002)
17/
FINAL
97
96­
hourEC50
(
cell
density)
=
71
(
66
­
73)
mg/
L
96­
hourEbC50
(
area
under
curve)
=
71
(
67­
74)
mg/
L
96­
hour
ErC50
(
growth
rate)
=
126
(
115
­
138)
mg/
L
96­
hour
EC90
(
cell
density)
=
137
(
105
­
153)
mg/
L
96­
hour
EbC90
(
area
under
curve)
=
145
(
125
­
155)
mg/
L
96­
hour
ErC90
(
growth
rate)
=
>
179
mg/
L
(
C.
I.
not
calculable)
72­
hour
NOEC
(
growth
rate,
cell
density,
area
under
the
curve)
=
44
mg/
L
96­
hour
NOEC
(
growth
rate,
cell
density,
area
under
the
curve)
=
44
mg/
L
All
element
values
based
on
mean
measured
concentrations
Statistical
methods:
Cell
densities,
area
under
the
growth
curve
values,
growth
rates
and
percent
inhibition
values
were
calculated
using
"
The
SAS
System
for
Windows",
Release
6.12.
These
values
were
then
analyzed
by
linear
interpolation
using
TOXSTAT
Version
3.5
to
estimate
the
ECIO,
EC50,
and
EC90
values
and
95%
confidence
limits
at
72
and
96
hours.
Cell
densities,
areas
under
the
growth
curve
and
growth
rates
at
72
and
96
hours
were
also
evaluated
for
normality
and
homogeneity
of
variances
using
the
Shapiro­
Wilks's
test
and
Bartlett's
test,
respectively.
The
treatment
groups
were
then
compared
to
the
control
using
Dunnett's
test.
Results
of
the
statistical
analyses
were
used
to
determine
the
NOEC
values.

Analytical
Methodology:
Analyses
of
test
solutions
were
performed
at
Wildlife
International
Ltd.
using
high
performance
liquid
chromatography
with
mass
spectrometric
detection
(
HPLC/
MS).
When
determining
the
concentration
of
the
test
substance
in
the
test
solutions,
the
same
and
most
prominent
peak
response
for
perfluorooctanesulfonate
was
used.
No
attempt
was
made
to
quantify
on
the
basis
of
individual
isomeric
components.
The
LOQ
(
limit
of
quantitation)
was
0.115
mg/
L
in
this
study.
The
mean
percent
recovery
of
matrix
fortifications
analyzed
concurrently
during
sample
analysis
was
99.1.
Samples
collected
at
test
initiation
had
measured
values
from
82.0
to
98.1%
of
nominal.
Measured
values
for
samples
taken
at
72
hours
ranged
from
91.7
to
105%
of
nominal.
Measured
values
for
samples
taken
at
96
hours
ranged
from
90.3
to
102%
of
nominal.
For
the
abiotic
controls,
measured
values
for
samples
taken
at72­
hours
ranged
from
81.9
to
105%
of
nominal
and
for
samples
taken
at
96­
hours,
90.3
to
103%
of
nominal.

Summary
of
analytical
chemistry
data:

Nominal
test
concentration,
mg/
l
Measured
values
at
0,
72,
and
96­
hours
respectively,
mg/
l
Mean
measured
concentration,
mg/
l
Percent
nominal
Negative
control
All
<
LOQ
<
LOQ
­
5.7
4.73,
6.04,
5.84
5.5
96
11
10.7,
11.2,
12.1
11
100
23
19.8,
23.1,
20.7
21
91
46
42.7,
41.9,
46.3
44
96
91
83.3,
86.0,
88.3
86
95
183
179,
186,
172
179
98
183
(
abiotic)
Not
analyzed,
150,
188
169
92
Control
response:
satisfactory
ENV/
JM/
RD(
2002)
17/
FINAL
98
Biological
observations
after
96­
hours:

Mean
measured
concentration,
mg/
l
Mean
number
of
cells
per
ml
Percent
Inhibition
via
Density
Percent
Inhibition
via
Area
Under
the
Curve
Percent
Inhibition
via
Growth
Rate
Negative
control
2,740,00
­
­
­
5.5
3,040,000
­
11
­
8.5
­
1.9
11
2,880,000
­
5.1
­
3.3
­
0.84
21
3,240,000
­
18
­
13
­
3.0
44
3,080,000
­
12
­
5.3
­
2.0
86
626,667
77
75
27
179
33,667
99
98
79
Observations:
After
96
hours
of
exposure,
there
were
no
signs
of
aggregation,
flocculation
or
adherence
of
the
algae
to
the
flasks
in
the
negative
control
or
any
test
treatment
group.
In
addition,
there
were
no
noticeable
changes
in
cell
color
or
morphology
when
compared
to
the
negative
control,
although
a
few
cells
appeared
enlarged
in
the
86
and
179
mg/
L
treatment
groups.

Reversibility
of
Growth
Inhibition:
The
179
mg/
L
treatment
group
was
maximally
inhibited
after
96­
hours.
Aliquots
of
the
test
solution
were
diluted
with
algal
medium
and
cultured
for
five
days.
Based
on
the
growth
observed
in
the
recovery
phase,
the
effect
on
algal
growth
was
found
to
be
algistatic.

CONCLUSIONS
The
potassium
perfluorooctanesulfonate
96­
hour
EC50
and
95%
confidence
interval
for
Selenastrum
capficornutumwas
determined
using
three
calculation
methods.
By
cell
density,
it
was
71
(
66
­
73)
mg/
L,
by
area
under
the
growth
curve
it
was
71
(
67
­
74)
mg/
L
and
by
growth
rate
126
(
115
­
138)
mg/
L.
The
96­
hour
NOEC
was
determined
by
Dunnett's
procedure
(
p
<
0.05)
to
be
44
mg/
L
using
all
three
methods.
No
signs
of
aggregation,
flocculation,
or
adherence
were
noted
in
any
of
the
test
solutions
or
the
controls.
This
test
substance
was
determined
to
be
algistatic.

Submitter:
3M
Company,
Environmental
Laboratory,
P.
O.
Box
33331,
St.
Paul,
Minnesota,
55133,
USA
DATA
QUALITY
Reliability:
Klimisch
ranking
1
REFERENCES
This
study
was
conducted
at
Wildlife
International
Ltd.,
Easton,
MD
at
the
request
of
the
3M
Company.

OTHER
Last
changed:
5/
3/
00
ENV/
JM/
RD(
2002)
17/
FINAL
99
Robust
Study
Report
Reference
No.
3
­
48­
Hour
Static
Acute
Toxicity
Test
with
the
Cladoceran
(













)

TEST
SUBSTANCE
Identity:
Potassium
perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
The
test
substance
is
a
white
powder.
Sample
was
taken
from
3M
lot
number
217.
Sample
was
stored
under
ambient
conditions
prior
to
testing.
Purity
determined
to
be
90.49%
by
LC/
MS,
1H­
HMR,
19FNMR
and
elemental
analyses
techniques.

METHOD
Method:
OECD
202
and
OPPTS
850.1010
Test
type:
Static
acute
GLP:
Yes
Year
completed:
Study
completed
1999.
Report
completed
2000
Species:
Daphnia
magna
Analytical
monitoring:
PFOS
measured
at
0,
24,
48­
hours
Statistical
methods:
EC50
values
calculated,
when
possible,
by
probit
analysis,
moving
average
method
or
binomial
probability
with
non­
linear
interpolation
using
the
computer
software
of
C.
E.
Stephan.
Test
daphnid
source:
Obtained
from
cultures
maintained
by
Wildlife
International
Ltd.,
Easton,
MD.
Identification
of
the
original
brood
stock
was
verified
by
the
Academy
of
Natural
Sciences,
Philadelphia,
PA.
,
USA
Test
daphnid
age
at
study
initiation:
<
24­
hours
Test
conditions
Dilution
water:
0.45
µ
m
filtered
well
water
Dilution
water
chemistry
(
during
the
4­
week
period
immediately
preceding
the
test):
Hardness:
132
(
128­
136)
mg/
L
as
CaCO3
Alkalinity:
178
(
176­
178)
mg/
L
as
CaCO3
pH:
8.3
(
8.2­
8.3)
TOC:
<
1.0
mg/
L
Conductivity:
313
(
310­
315)
µ
mhos/
cm
Ca/
Mg
ratio:
35/
13.5
Na/
K
ratio:
21.3/
6.62
Lighting:
Colortone
®
50
fluorescent
lights,
intensity
approximately
359
lux.
Photoperiod
of
16­
hours
light,
8­
hours
dark
with
a
30­
minute
transition
period.
Stock
and
test
solutions
preparation:
A
primary
stock
solution
was
prepared
in
dilution
water
at
91
mg/
L.
It
was
mixed
for
~
19.5
hours
prior
to
use.
After
mixing,
the
primary
stock
was
proportionally
diluted
with
dilution
water
to
prepare
the
four
additional
test
concentrations.
All
test
solutions
appeared
clear
and
colorless.
Exposure
vessels:
250
mL
plastic
beakers
containing
240
mL
of
test
solution.
The
approximate
depth
of
test
solution
was
6.4
cm.
Number
of
replicates:
two
Number
of
daphnids
per
replicate:
ten
Number
of
concentrations:
five
plus
a
negative
control
Water
chemistry
during
the
study:
Dissolved
oxygen
range
(
0
 
48
hours):
8.6
 
8.9
mg/
L
(
control
exposure)
ENV/
JM/
RD(
2002)
17/
FINAL
100
8.6
 
9.1
mg/
L
(
91
mg/
L
exposure)
pH
range
(
0
 
48
hours)
8.2
 
8.5
(
control
exposure)
8.5
 
8.6
(
91
mg/
L
exposure)
Test
temperature
range
(
0
 
48
hours)
19.5
 
20.2oC
(
control
exposure)
19.3
 
20.1oC
(
91
mg/
L
exposure)
Element
basis:
mortality
and
immobilization
Method
of
calculating
mean
measured
concentrations:
arithmetic
mean
RESULTS
Nominal
concentrations:
Negative
control,
12,
20,
33,
55,
91
mg/
L
Measured
concentrations:
<
LOQ,
11,
20,
33,
56,
91
mg/
L
Element
value:
(
95%
confidence
interval
is
given
in
brackets)
24­
hour
EC10
=
82
(
81­
83)
mg/
L
24­
hour
EC50
=
>
91
mg/
L
(
C.
I.
not
calculable)
24­
hour
EC90
=
>
91
mg/
L
(
C.
I.
not
calculable)
48­
hour
EC10
=
53
(<
11­>
91)
mg/
L
48­
hour
EC50
=
61
(
33­
91)
mg/
L
48­
hour
EC90
=
63
(<
11­>
91)
mg/
L
All
element
values
based
on
mean
measured
concentrations
Statistical
evaluation:
The
EC50
values
and
95%
confidence
intervals
were
calculated
when
possible
by
probit
analysis,
the
moving
average
method
or
binomial
probability
with
non­
linear
interpolation
using
the
computer
software
of
C.
E.
Stephan.
The
EC10
and
EC90
values
were
calculated
when
possible
using
the
Bruce­
Versteeg
method
because
there
were
less
than
two
concentrations
with
partial
mortality
or
immobility.
Analytical
methodology:
Analyses
of
test
solutions
were
performed
at
Wildlife
International
Ltd.
using
high
performance
liquid
chromatography
with
mass
spectrometric
detection
(
HPLC/
MS).
When
determining
the
concentration
of
the
test
substance
in
the
test
solutions,
the
same
and
most
prominent
peak
response
for
perfluorooctanesulfonate
was
used.
No
attempt
was
made
to
quantify
on
the
basis
of
individual
isomeric
components.
The
LOQ
(
limit
of
quantitation)
was
4.58
mg/
L
in
this
study.
The
mean
percent
recovery
of
matrix
fortifications
analyzed
concurrently
during
sample
analysis
was
96.2.
Samples
collected
at
test
initiation
had
measured
values
from
85.5
to
112%
of
nominal.
Measured
values
for
samples
taken
at
24
hours
ranged
from
92.2
to
115%
of
nominal.
Measured
values
for
samples
taken
at
48
hours
ranged
from
91.6
to
106%
of
nominal.

Summary
of
analytical
chemistry
data:

Nominal
Test
Concentration
mg/
L
Measured
duplicated
values
at
0,
24,
and
48­
hours
respectively,
mg/
L
Mean
Measured
Concentration
mg/
L
Percent
Of
Nominal
Negative
Control
All
<
LOQ
<
LOQ
­
12
10.5,
10.6,
11.5,
12.5,
10.9,
12.0
11
92
20
17.2,
18.1,
22.8,
21.6,
21.4,
18.8
20
100
33
30.2,
34.1,
34.0,
36.1,
31.3,
34.0
33
100
55
50.5,
49.9,
57.0,
63.0,
56.8,
56.4
56
102
91
87.6,
102,
90.1,
84.4,
88.7,
92.4
91
100
Biological
observations
after
48­
hours:
Daphnids
in
the
negative
control,
the
11
and
the
20
mg/
L
ENV/
JM/
RD(
2002)
17/
FINAL
101
treatments
appeared
healthy
and
normal
throughout
the
test
with
no
mortality,
immobility
or
overt
clinical
signs
of
toxicity.
Five
percent
mortality
was
observed
at
48­
hours
in
the
negative
control.
The
effects
noted
in
this
study
were
mortality;
no
immobilization
was
noted
at
any
test
concentration.

Cumulative
percent
mortality:

Mean
Measured
Test
Concentration
mg/
L
24­
hours
48­
hours
Negative
Control
0
5
11
0
0
20
0
0
33
0
0
56
0
35
91
35
100
Control
response:
satisfactory
CONCLUSIONS
The
potassium
perfluorooctanesulfonate
48­
hour
EC50
for
Daphnia
magna
was
determined
to
be
61
mg/
L
with
a
95%
confidence
interval
of
33­
91
mg/
L.
The
48­
hour
no
immobilization
and
no
observed
effect
concentration
was
33
mg/
L.

Submitter:
3M
Company,
Environmental
Laboratory
P.
O.
Box
33331
St.
Paul,
MN
55133,
USA
DATA
QUALITY
Reliability:
Klimisch
ranking
1
REFERENCES
This
study
was
conducted
at
Wildlife
International,
Ltd.
Easton,
MD
at
the
request
of
the
3M
Company.

OTHER
Last
changed:
5/
3/
00
ENV/
JM/
RD(
2002)
17/
FINAL
102
Robust
Study
Report
Reference
No.
4
­
96­
Hour
Shell
Deposition
Test
with
the
Eastern
Oyster
(





















)

TEST
SUBSTANCE
Identity:
Potassium
pertluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
The
test
substance
is
a
white
powder.
Sample
was
taken
from
3M
lot
number
217.
Sample
was
stored
under
ambient
conditions
prior
to
testing.
Purity
determined
to
be
90.49%
by
LC/
MS,
1H­
HMR,
19FNMR
and
elemental
analyses
techniques.

METHOD
Method:
OPPTS
850.1025
Type:
Static
acute
GLP:
Yes
Year
completed:
Study
completed
1999.
Report
completed
2000
Species:
Crassostrea
virginica
Supplier:
P.
Cummins
Oyster
Company,
Inc.,
Baltimore,
MD,
USA
Shell
grinding:
Prior
to
test
initiation,
recently
deposited
shell
at
the
rounded
(
ventral)
end
was
removed
using
a
small
electric
grinder.
Care
was
taken
to
remove
the
shell
rim
uniformly
to
produce
a
smooth,
rounded,
blunt
profile.
Analytical
monitoring:
PFOS
measured
at
0,48,96­
hours
Exposure
period:
96­
hours
Statistical
methods:
Shell
growth
inhibition
was
calculated
for
each
treatment
group
as
the
percent
reduction
in
shell
growth
relative
to
mean
shell
growth
in
the
negative
control.
The
EC50
value
was
estimated
by
visual
inspection
of
shell
growth
inhibition
data.
The
shell
growth
data
was
evaluated
for
normality
and
homogeneity
of
variances
using
the
Chi­
Square
test
and
Bartlett's
test,
respectively.
Dunnett's
test
was
used
to
identify
treatment
groups
that
had
a
statistically
significant
(<
0.05)
reduction
in
shell
growth
as
compared
to
the
control.
Test
oyster
age:
unknown
Length:
33.8
(
27.8­
41.5)
mm,
Pretreatment:
None
Test
conditions:
Dilution
water:
Natural
seawater
diluted
to
a
salinity
of
20
O/
oo
with
well
water
Dilution
water
chemistry
(
during
the
4­
week
period
immediately
preceding
the
test):
salinity:
21
(
20­
21)
O/
oo
pH:
8.1(
8.0­
8.2)
TOC:
<
1.0
mg/
L
Stock
and
test
solution
preparation:
Primary
stock
prepared
in
dilution
water
at
9.1mg/
L
and
mixed
for
~
24
hours
prior
to
use.
After
mixing,
primary
stock
solution
appeared
clear
and
colorless
with
some
white
particulate
material
suspended
throughout
the
solution.
It
was
proportionally
diluted
with
dilution
water
to
prepare
the
four
additional
test
concentrations.
All
test
solutions
appeared
clear
and
colorless.
Due
to
the
relatively
low
solubility
of
PFOS
in
natural
seawater,
the
highest
concentration
attainable
with
this
matrix
is
approximately
3.3mg/
L.
Concentrations
dosing
rate:
Once
Exposure
vessels:
52L
polyethylene
aquaria
containing
approximately
40L
of
test
solution;
water
depth
approximately
21
cm.
Each
chamber
was
continuously
stirred
to
circulate
the
supplemental
ENV/
JM/
RD(
2002)
17/
FINAL
103
algae
diet
using
an
electric
paddle
mixer.
Feeding:
Algal
cells
(
Thalassiosira
pseudonana,
Skeletonema
sp.,
Chaetoceros
sp.,
and
lsochrysis
sp.)
were
provided
to
supplement
naturally
occurring
algae
and
to
maximize
oyster
growth
rates
during
the
test.
Number
of
replicates:
one
Number
of
oysters
per
replicate:
twenty
Number
of
concentrations:
five
plus
a
negative
control
Water
chemistry
during
the
study:
Dissolved
oxygen
range
(
0­
96
hours):
6.6
­
7.6mg/
L
(
control
exposure)
6.1
­
7.7mg/
L
(
3.0
mg/
L
exposure).
pH
range
(
0­
96
hours):
7.6
­
8.1
(
control
exposure)
7.6
­
8.1
(
3.0
mg/
L
exposure)
Test
temperature
range
(
0­
96
hours):
22.2
­
22.3
oC
(
control
exposure)
21.8
­
22.7
oC
(
3.0
mg/
L
exposure)
Method
of
calculating
mean
measured
concentrations:
arithmetic
mean
RESULTS
Nominal
concentrations:
Negative
control,
1.2,
2.0,
3.3,
5.5,
9.1
mg/
L
Measured
concentrations:
<
LOQ,
0.36,
0.40,
1.3,
1.9,
3.0
mg/
L
Element
value:
(
95%
confidence
interval
is
given
in
brackets)
96­
hour
EC50
=
>
3.0
mg/
L
(
C.
I.
not
calculable)
96­
hour
NOEC
=
1.9
mg/
L
All
element
values
based
on
mean
measured
concentrations
Statistical
evaluation
of
shell
growth:
EC50
values
could
not
be
calculated
due
to
insufficient
shell
growth
inhibition
at
the
highest
attainable
concentration.

Analytical
Methodology:
Analyses
of
test
solutions
were
performed
at
Wildlife
International
Ltd.
using
high
performance
liquid
chromatography
with
mass
spectrometric
detection
(
HPLC/
MS).
When
determining
the
concentration
of
the
test
substance
in
the
test
solutions,
the
same
and
most
prominent
peak
response
for
perfluorooctanesulfonate
was
used.
No
attempt
was
made
to
quantify
on
the
basis
of
individual
isomeric
components.
The
LOQ
(
limit
of
quantitation)
was
0.115
mg/
L
in
this
study.
Samples
collected
at
test
initiation
had
measured
values
from
28
to
46%
of
nominal.
Measured
values
for
samples
taken
at
48­
hours
ranged
from
15
to
41
%
of
nominal.
Measured
values
for
samples
taken
at
96­
hours
ranged
from
<
LOQ
to
52%
of
nominal.

Summary
of
analytical
chemistry
data:

Nominal
Test
Concentration,
mgll
Measured
Duplicate
Values
at
0,
48
and
96­
hours,
Respectively,
mg/
L
Mean
Measured
Concentration,
mg/
l
Percent
of
Nominal
Negative
Control
All
<
LOQ
<
LOQ
­
1.2
0.331,
0.353,
0.341,
0.429,
<
LOQ,
<
LOQ
0.36
30
2.0
0/
622,
0.633,
0.299,
0.40
20
ENV/
JM/
RD(
2002)
17/
FINAL
104
0.313,
0.249,
0.257
3.3
1.36,
1.15,
0.924,
0.878,
1.58,
1.72
1.3
39
5.5
2.42,
2.53,
2.02,
2.24,
1.45,
0.970
1.9
35
9.1
3.39,
3.44,
3.01*,
3.74,
3.57,
1.99,
2.19
3.0
33
*
3
replicates
analyzed
at
time
0
Biological
observations
after
96­
hours:
Oysters
in
the
negative
control
and
all
PFOS
treatment
groups
appeared
normal
and
healthy
throughout
the
exposure
period.

Shell
deposition
and
shell
growth
Inhibition
at
test
termination:

Mean
Measured
Concentration,
mg/
l
Shell
Deposition
Mean
+
SD,
mm
Percent
Inhibition
in
Shell
Growth
Negative
Control
2.67
+
0.824
­
0.36
2.50
+
0.933
6.4
0.40
2.40
+
0.820
10
1.3
2.51
+
0.919
6.0
1.9
2.13
+
0.804
20
3.0
1.91
+
0.591
28
Mortality
of
controls:
None
CONCLUSIONS
The
potassium
perfluorooctanesulfonate
96­
hour
EC50
for
the
Eastern
Oyster
was
determined
to
be
>
3.0
mg/
L,
the
highest
concentration
tested
and
the
practical
limit
of
solubility
in
unfiltered
seawater.
The
96­
hour
no
effect
concentration
was
1.9
mg/
L.

Submitter:
3M
Company,
Environmental
Laboratory,
P.
O.
Box
33331,
St.
Paul,
Minnesota,
55133,
USA
DATA
QUALITY
Reliability:
Klimisch
ranking
=
1
REFERENCES
This
study
was
conducted
at
Wildlife
International
Ltd.,
Easton,
MD
at
the
request
of
the
3M
Company.

OTHER
Last
changed:
5/
3100
ENV/
JM/
RD(
2002)
17/
FINAL
105
Robust
Study
Report
Reference
No.
5
­
96­
Hour
Static
Acute
Toxicity
Test
with
the
Freshwater
Mussel
(









	






)

TEST
SUBSTANCE
Identity:
Potassium
perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
The
test
substance
is
a
white
powder.
Sample
was
taken
from
3M
lot
number
217.
Sample
was
stored
under
ambient
conditions
prior
to
testing.
Purity
determined
to
be
90.49%
by
LC/
MS,
1H­
HMR,
19FNMR
and
elemental
analyses
techniques.

METHOD
Method:
The
study
was
conducted
using
a
protocol
based
on
procedures
outlined
in
U.
S.
Environmental
Protection
Agency
Series
850
 
Ecological
Effects
Guidelines,
OPPTS
Number
850.1075
;
OECD
203:
Fish,
Acute
Toxicity
Test;
and
ASTM
Standard
E729­
88a,
Standard
Guide
for
Conducting
Toxicity
Tests
with
Fishes,
Macroinvertebrates
and
Amphibians.
Test
type:
Semi­
static
Renewal
GLP:
Yes
Year
completed:
Study
completed
1999.
Report
completed
2000
Species:
Unio
complamatus
Analytical
monitoring:
Test
substance
concentrations
measured
by
LCMS
at
0,
48,
96­
hours
Statistical
methods:
LC50
values
calculated,
when
possible,
by
probit
analysis,
moving
average
method
or
binomial
probability
with
non­
linear
interpolation
using
the
computer
software
of
C.
E.
Stephan.
Test
organism
source:
Obtained
from
Carolina
Biological
Supply
Company,
Burlington,
North
Carolina,
USA.
Carolina
collected
from
the
wild.
Test
organism
age
at
study
initiation:
Unknown
Test
Conditions
Dilution
water:
0.45
µ
m
filtered
well
water
Dilution
water
chemistry
(
during
the
4­
week
period
immediately
preceding
the
test):
Hardness:
126
(
120­
132)
mg/
L
as
CaCO3
Alkalinity:
174
(
170­
178)
mg/
L
as
CaCO3
pH:
8.3
(
8.1­
8.5)
TOC:
<
1.0
mg/
L
Conductivity:
321
(
310­
330)
µ
mhos/
cm
Ca/
Mg
ratio:
35/
13.5
Na/
K
ratio:
21.3/
6.62
Lighting:
Colortone
®
50
fluorescent
lights,
intensity
approximately
369
lux.
Photoperiod
of
16­
hours
light,
8­
hours
dark
with
a
30­
minute
transition
period.
Stock
and
test
solutions
preparation:
A
primary
stock
solution
was
prepared
in
dilution
water
at
91
mg/
L.
It
was
mixed
for
approximately
24
hours
prior
to
use.
After
mixing,
the
primary
stock
was
proportionally
diluted
with
dilution
water
to
prepare
the
four
additional
test
concentrations.
All
test
solutions
appeared
clear
and
colorless.
Exposure
vessels:
25
liter
polyethylene
aquaria
containing
approximately
20
L
of
test
solution.
The
approximate
depth
of
test
solution
was
23.2
cm.
Number
of
replicates:
two
Number
of
test
organisms
per
replicate:
ten
Number
of
concentrations:
five
plus
a
negative
control
Water
chemistry
during
the
study:
ENV/
JM/
RD(
2002)
17/
FINAL
106
Dissolved
oxygen
range
(
0
 
96
hours):
5.8
 
8.5
mg/
L
(
control
exposure)
5.0
 
8.6
mg/
L
(
79
mg/
L
exposure)
pH
range
(
0
 
96
hours):
8.0
 
8.4
(
control
exposure)
7.9
 
8.5
(
79
mg/
L
exposure)
Test
temperature
range
(
0
 
96
hours):
21.4
 
21.8oC
(
control
exposure)
21.8
 
23.7
oC
(
79
mg/
L
exposure)
Element
Basis:
Mortality.
Mussels
with
open
shells
and
not
responding
to
gentle
prodding
were
considered
dead.
The
number
of
individuals
exhibiting
clinical
signs
of
toxicity
or
abnormal
behavior
also
were
evaluated.
Method
of
calculating
mean
measured
concentrations:
arithmetic
mean
RESULTS
Nominal
concentrations:
<
LOQ,
5.7,
11,
23,
46,
91
mg/
L
Measured
concentrations:
<
LOQ,
5.3,
12,
20,
41,
79
mg/
L
Element
value:
(
95%
confidence
interval
is
given
in
brackets)
96­
hour
LC50
=
59
mg/
L
(
51­
68
mg/
L)

Statistical
evaluation:
The
LC50
values
and
95%
confidence
intervals
were
calculated
when
possible
by
probit
analysis,
the
moving
average
method
or
binomial
probability
with
non­
linear
interpolation
using
the
computer
software
of
C.
E.
Stephan.

Analytical
methodology:
Analyses
of
test
solutions
were
performed
at
Wildlife
International
Ltd.
using
high
performance
liquid
chromatography
with
mass
spectrometric
detection
(
HPLC/
MS).
When
determining
the
concentration
of
the
test
substance
in
the
test
solutions,
the
same
and
most
prominent
peak
response
for
perfluorooctanesulfonate
was
used.
No
attempt
was
made
to
quantify
on
the
basis
of
individual
isomeric
components.
The
LOQ
(
limit
of
quantitation)
was
0.115
mg/
L
in
this
study.
The
mean
percent
recovery
of
matrix
fortifications
analyzed
concurrently
during
sample
analysis
was
94.7%.
Samples
collected
at
test
initiation
had
measured
values
from
73.7%
to
96.0%
of
nominal.
Measured
values
for
samples
taken
at
48
hours
ranged
from
81.2
to
98.9%
of
nominal.
Measured
values
for
samples
taken
at
96
hours
ranged
from
88.5
to
130%
of
nominal.

Summary
of
analytical
chemistry
data:

Nominal
Test
Concentration
,
mg/
L
Measured
duplicated
values
at
0,
48,
and
96­
hours
respectively,
mg/
L
Mean
Measured
Concentration,
mg/
L
Percent
of
Nominal
Negative
Control
All
<
LOQ
<
LOQ
­
5.7
5.47,
4.93,
5.18,
5.70,
5.24,
5.26
5.3
93
11
11.4,
10.1,
11.2,
10.5,
10.9,
15.4
12
109
23
19.0,
16.8,
18.7,
18.7,
22.9,
22.4
20
87
46
37.2,
40.6,
37.1,
39.5,
48.2,
40.5
41
89
91
69.0,
74.7,
81.3,
77.6,
88.2,
85.7
79
87
Biological
observations
after
96­
hours:
Mussels
in
the
negative
control,
the
5.3,
12
and
the
20
mg/
L
treatments
appeared
healthy
and
normal
throughout
the
test
with
no
mortality
or
overt
clinical
signs
of
toxicity.
Five
percent
mortality
was
observed
at
96­
hours
in
the
41
mg/
L
treatment
and
90%
mortality
was
observed
in
the
79
mg/
L
treatment.
No
abnormal
behavior
was
noted
in
these
concentrations.
ENV/
JM/
RD(
2002)
17/
FINAL
107
Cumulative
percent
mortality:

Mean
Measured
Test
Concentration
mg/
L
24
Hours
48
Hours
72
Hours
96
Hours
Negative
Control
0
0
0
0
5.3
0
0
0
0
12
0
0
0
0
20
0
0
0
0
41
0
0
0
5
79
30
40
50
90
Control
response:
Satisfactory
CONCLUSIONS
The
potassium
perfluorooctanesulfonate
96­
hour
LC50
for
the
Freshwater
Mussel,
Unio
complamatus
was
determined
to
be
59
mg/
L
with
a
95%
confidence
interval
of
51­
68
mg/
L.
The
96­
hour
no
mortality
concentration
was
20
mg/
L.

Submitter:
3M
Corporation,
Environmental
Laboratory
P.
O.
Box
33331
St.
Paul,
MN
55133,
USA
DATA
QUALITY
Reliability:
Klimisch
ranking
1.

REFERENCES
This
study
was
conducted
at
Wildlife
International,
Ltd.
Easton,
MD
at
the
request
of
the
3M
Company.

OTHER
Last
changed:
5/
3/
00
ENV/
JM/
RD(
2002)
17/
FINAL
108
Robust
Study
Report
Reference
No.
6
 
Activated
Sludge,
Respiration
Inhibition
Test
TEST
SUBSTANCE
Identity:
Potassium
perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt
CAS
#
2795­
39­
3)

Remarks:
The
test
substance
is
a
white
powder.
Sample
was
taken
from
3M
lot
number
217.
Sample
was
stored
under
ambient
conditions
prior
to
testing.
Purity
determined
to
be
90.49%
by
LC/
MS,
1H­
HMR,
19F­
NMR
and
elemental
analyses
techniques.

METHOD
Method:
OECD
209
Test
type:
Static
acute
GLP:
Yes
Year
Completed:
Study
completed
1999.
Report
completed
2000
Analytical
monitoring:
Dissolved
oxygen
concentrations.
Statistical
methods:
Probit
analysis
using
the
computer
software
of
C.
E.
Stephan.
Test
organism
source.
Activated
sludge
collected
from
the
Prospect
Bay
Wastewater
Treatment
Facility,
Grasonville,
Maryland,
USA.

Test
conditions
Dilution
water:
NANOpure
®
Synthetic
Sewage:
1
liter
municipal
water
16.0
g
peptone
11.0
g
meat
extract
3.0
g
urea
0.7
g
NaCl
0.4
g
CaCI2
2H20
0.2
g
MgSO4
7H20
2.8
g
K2HPO4
Reference
and
test
solution
preparation:
A
stock
solution
of
the
reference
substance,
3,5­
dichlorophenol,
was
prepared
by
dissolving
500
mg
in
10
mL
of
1N
NaOH,
diluted
to
30
mL
with
NANOpure
 
water,
then
brought
to
the
point
of
incipient
precipitation
with
1
NH2S04,
and
diluted
to
1
L
with
NANOpure4
 
water.
The
pH
of
the
reference
solution
was
measured
to
be
7.18.
PFOS
was
added
directly
to
test
vessels
rather
than
volumetric
addition
of
a
stock
solution.
This
method
was
deemed
appropriate
based
on
the
observed
solubility
of
the
test
substance
in
water.
Test
vessels:
Mixtures
were
prepared
and
aerated
in
500
mL
Erlenmeyer
flasks
and
then
transferred
into
300
mL
Biochemical
Oxygen
Demand
(
BOD)
bottles
Number
of
concentrations:
7
plus
3
reference
controls
and
2
Blank
controls
Temperature:
19­
21oC
Total
Suspended
Solids
and
pH
for
sludge
on
day
of
testing:
4380
mg/
L
and
7.87
respectively.
Element
Basis:
Respiration
inhibition
as
determined
by
dissolved
oxygen
concentration.

Method
Remarks:
Stock
solutions
of
PFOS
that
were
prepared
at
nominal
concentration
of
approximately
500
and
1
000
mg/
L
in
NANOpure
 
water
contained
test
material
that
was
not
in
solution
after
20­
minutes
of
sonication.
Therefore,
direct
weight
addition
was
employed
to
administer
PFOS
to
the
test
system.
ENV/
JM/
RD(
2002)
17/
FINAL
109
Test
mixtures
were
prepared
at
15­
minute
intervals
and
aerated
until
the
contact
time
of
the
test
substance
with
the
activated
sludge
was
three
hours.
After
3­
hours
of
contact
time,
dissolved
oxygen
was
measured
over
a
period
of
up
to
10­
minutes.

RESULTS
Nominal
concentrations:
Two
blank
controls,
three
reference
substance
controls,
0.90,
2.7,
9.0,
27,
90,
271.
905
mg/
L
test
material
solutions.

Statistical
Analyses:
EC50
values
were
calculated
for
the
reference
material
by
probit
analysis
using
the
computer
software
of
C.
E.
Stephan.
An
EC50
value
could
not
be
calculated
for
the
test
substance.

Analytical
Methodology:
Analysis
of
DO
concentrations
in
all
test
solutions
were
performed
at
Wildlife
International
Ltd.
using
a
YSI
Model
50B
Dissolved
Oxygen
Meter.
Dissolved
oxygen
readings
were
recorded
every
10
seconds
for
10
minutes
or
until
the
dissolved
oxygen
dropped
below
1.0
mg/
L
Respiration
Rates
and
Percent
Inhibitions
Treatment
Respiration
Rate
mg
02/
L/
hour
Percent
Inhibition
Control
1
39.6
NA
Control
2
41.1
NA
3,5­
dichlorophenol
3mg/
L
31.1
22.9
3,5­
dichlorophenol
15
mg/
L
14.6
63.8
3,5­
dichlorophenol
50
mg/
L
5.1
87.4
Test
substance
0.90
mg/
L
38.9
3.6
Test
substance
2.7
mg/
L
35.1
13.0
Test
substance
9.0
mg/
L
33.5
17.0
Test
substance
27
mg/
L
37.9
6.1
Test
substance
90
mg/
L
32.7
19.0
Test
substance
271
mg/
L
28.1
30.4
Test
substance
905
mg/
L
24.7
38.8
Control
response:
satisfactory
CONCLUSIONS
The
test
substance
exhibited
a
maximum
inhibitory
effect
of
38%
upon
respiration
at
a
nominal
test
substance
concentration
of
905
mg/
L.
The
EC50
(
respiration
inhibition)
is
therefore
greater
than
the
solubility
of
the
test
substance.

Submitter:
3M
Company,
Environmental
Laboratory,
P.
O.
Box
33331,
St.
Paul,
Minnesota,
55133,
USA
DATA
QUALITY
Reliability:
Klimisch
ranking
1.

REFERENCES
This
study
was
conducted
at
Wildlife
International,
Ltd.
Easton,
MD
at
the
request
of
the
3M
Company.
ENV/
JM/
RD(
2002)
17/
FINAL
110
OTHER
Last
changed:
5/
3/
00
ENV/
JM/
RD(
2002)
17/
FINAL
111
Robust
Study
Report
Reference
No.
7
­
96­
Hour
Static
Acute
Toxicity
Test
with
the
Saltwater
Mysid
(
















)

TEST
SUBSTANCE
Identity:
Potassium
perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
The
test
substance
is
a
white
powder.
Sample
was
taken
from
3M
lot
number
217.
Sample
was
stored
under
ambient
conditions
prior
to
testing.
Purity
determined
to
be
90.49%
by
LC/
MS,
1H­
HMR,
19F­
NMR
and
elemental
analyses
techniques.

METHOD
Method:
OPPTS
850.1035
Type:
Static
acute
GLP:
Yes
Year
completed:
Study
completed
1999.
Report
completed
2000
Species:
Mysidopsis
bahia
Supplier:
In­
house
cultures,
Wildlife
International,
Ltd.,
Easton,
MD,
USA
Analytical
monitoring:
PFOS
measured
at
0,
48,
96­
hours
Exposure
period:
96­
hours
Statistical
methods:
LC50
values
calculated,
when
possible,
by
probit
analysis,
moving
average
method
or
binomial
probability
with
non­
linear
interpolation
using
the
computer
software
of
C.
E.
Stephan.
Test
fish
age:
<
24­
hours
old
Pretreatment:
None
Test
Conditions:
Dilution
water:
Natural
seawater
diluted
to
20%
o
with
well
water,
0.45
µ
m
filtered.
Dilution
water
chemistry
(
during
the
4­
week
period
immediately
preceding
the
test):
Salinity:
20
(
20­
20)
%
o
pH:
8.2
(
8.1­
8.2)
TOC:
<
1.0
mg/
L
Stock
and
test
solution
preparation:
Primary
stock
prepared
at
8.2
mg/
L
and
mixed
for
~
22
hours
prior
to
use.
After
mixing,
primary
stock
solution
was
proportionally
diluted
with
dilution
water
to
prepare
the
four
additional
test
concentrations.
All
test
solutions
appeared
clear
and
colorless.
Concentrations
dosing
rate:
Once
Stability
of
the
test
chemical
solutions:
Extremely
stable
Exposure
vessels:
2L
polyethylene
aquaria
containing
approximately
1000mL
of
test
solution;
water
depth
approximately
6.6
cm.
Number
of
replicates:
two
Number
of
mysids
per
replicate:
ten
Number
of
concentrations:
five
plus
a
negative
control
Feeding:
Live
brine
shrimp
nauplii
daily
Water
chemistry
during
the
study:
Dissolved
oxygen
range
(
0
 
96
hours):
6.8
 
7.4
mg/
L
(
control
exposure)
6.8
 
7.3
mg/
L
(
5.4
mg/
L
exposure)
pH
range
(
0
 
96
hours):
ENV/
JM/
RD(
2002)
17/
FINAL
112
8.1
 
8.2
(
control
exposure)
8.1
 
8.2
(
5.4
mg/
L
exposure)
Test
temperature
range
(
0
 
96
hours):
24.2
 
25.4oC
(
control
exposure)
23.8
 
24.5oC
(
5.4
mg/
L
exposure)
Method
of
calculating
mean
measured
concentrations:
arithmetic
mean
RESULTS
Nominal
concentrations:
Negative
control,
1.1,
1.8,
3.0,
4.9,
8.2
mg/
L
Measured
concentrations:
<
LOQ,
0.57,
1.1,
1.9,
3.0,
5.4
mg/
L
Element
value:
(
95%
confidence
interval
is
given
in
brackets)
24­
hour
LC50
=
>
5.4
mg/
L
(
CI
not
calculable)
48­
hour
LC50
=
>
5.4
mg/
L
C.
I.
not
calculable)
72­
hour
LC50
=
4.4
(
3.6­
6.2)
mg/
L
96­
hour
LC50
=
3.6
(
3.0­
4.6)
mg/
L
All
element
values
based
on
mean
measured
concentrations
Statistical
evaluation
of
mortality:
LC50
values
could
not
be
calculated
for
24
and
48­
hours
of
exposure
due
to
the
lack
on
an
adequate
concentration­
response
pattern.
The
probit
method
was
used
to
evaluate
mortality
at
72
and
96
hours.

Analytical
methodology:
Analyses
of
test
solutions
were
performed
at
Wildlife
International
Ltd.
using
high
performance
liquid
chromatography
with
mass
spectrometric
detection
(
HPLC/
MS).
When
determining
the
concentration
of
the
test
substance
in
the
test
solutions,
the
same
and
most
prominent
peak
response
for
perfluorooctane
sulfonate
was
used.
No
attempt
was
made
to
quantify
on
the
basis
of
individual
isomeric
components.
The
LOQ
(
limit
of
quantitation)
was
0.115
mg/
L
in
this
study.
The
mean
percent
recovery
of
matrix
fortifications
analyzed
concurrently
during
sample
analysis
was
97.4.
Samples
collected
at
test
initiation
had
measured
values
from
52.4
to
70.7%
of
nominal.
Measured
values
for
samples
taken
at
48
hours
ranged
from
43.5
to
71.0%
of
nominal.
Measured
values
for
samples
taken
at
96
hours
ranged
from
35.5
to
71.1%
of
nominal.

Summary
of
analytical
chemistry
data:

Nominal
Test
Concentration,
mg/
L
Measured
duplicate
values
at
0,
48,
and
96­
hours
respectively,
mg/
L
Mean
Measured
Concentration
mg/
L
Percent
of
Nominal
Negative
Control
All
<
LOQ
<
LOQ
­
1.1
0.575,
0.622,
0.605,
0.640,
0.391,
0.580
0.57
52
1.8
1.12,
1.19,
1.10,
1.09,
1.04,
1.13
1.1
61
3.0
1.92,
1.99,
1.92,
1.91,
1.79,
1.91
1.9
63
4.9
3.05,
2.66,
2.96,
3.35,
3.11,
3.11
3.0
61
8.2
5.82,
5.78,
3.58,
5.85,
5.22,
5.86
5.4
66
Biological
observations
after
96­
hours:
Mysids
in
the
negative
control,
and
the
0.57
and
1.1
mg/
L
(
mean
measured
concentrations)
treatment
groups
appeared
normal
and
healthy
during
the
test.
ENV/
JM/
RD(
2002)
17/
FINAL
113
Cumulative
percent
mortality:

Mean
Measured
Test
Concentration,
mg/
L
24­
hours
48­
hours
72­
hours
96­
hours
Negative
Control
0
0
0
0
0.57
0
0
0
0
1.1
0
0
0
0
1.9
0
0
5
10
3.0
5
15
30
40
5.4
15
45
60
75
Mortality
of
controls:
None
CONCLUSIONS
The
potassium
perfluorooctanesulfonate
96­
hour
LC50
for
saltwater
mysids
was
determined
to
be
3.6
mg/
L
with
a
95%
confidence
interval
of
3.0
 
4.6
mg/
L.
The
96­
hour
no
mortality
and
NOEC
concentration
was
1.1
mg/
L.

Submitter:
3M
Company,
Environmental
Laboratory
P.
O.
Box
33331
St.
Paul,
MN
55133,
USA
DATA
QUALITY
Reliability:
Klimisch
ranking
=
1
REFERENCES
This
study
was
conducted
at
Wildlife
International
Ltd.,
Easton,
MD
at
the
request
of
the
3M
Company.

OTHER
Last
changed:
5/
3/
00
ENV/
JM/
RD(
2002)
17/
FINAL
114
Robust
Study
Report
Reference
No.
8
­
Early
Life­
Stage
Toxicity
Test
with
the
Fathead
Minnow
(







	








	


)

TEST
SUBSTANCE
Identity:
Potassium
perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
The
test
substance
is
a
white
powder.
Sample
was
taken
from
3M
lot
number
217.
Sample
was
stored
under
ambient
conditions
prior
to
testing.
Purity
determined
to
be
90.49%
by
LC/
MS,
1H­
HMR,
19FNMR
and
elemental
analyses
techniques.

METHOD
Method:
OECD
210,
OPPTS
850.1400
Type:
Flow­
through
chronic
GLP:
Yes
Year
completed:
Study
completed
1999.
Report
completed
2000
Species:
Pimephales
promelas
Supplier:
In­
house
cultures,
Wildlife
International,
Ltd.,
Easton,
MD,
USA
Analytical
monitoring:
PFOS
measured
on
days
0,
4,
7,
14,
21,
28,
35,
42,
and
47
Exposure
period:
47
days
Statistical
methods:
Discrete­
variable
data
were
analyzed
using
2
x
2
contingency
tables
to
identify
treatment
groups
that
showed
a
statistically
significant
difference
(
p<
0.05)
from
the
negative
control
group.
All
continuous­
variable
data
were
evaluated
for
normality
using
Shapiro­
Wilk's
test
and
for
homogeneity
of
variance
using
Bartlett's
test.
Analysis
of
variance
and
Dunnett's
test
were
used
to
evaluate
differences
between
treatment
and
control
means.
Test
fish
age:
eggs
<
24­
hours
old
at
test
initiation
Pretreatment:
None
Test
Conditions
Dilution
water:
0.45
µ
m
filtered
well
water
Dilution
water
chemistry
(
during
the
4­
week
period
immediately
preceding
the
test):
Hardness:
126
(
124­
128)
mg/
L
as
CaCO3
Alkalinity:
172
(
170­
172)
mg/
L
as
CaCO3
pH:
8.2
(
8.2­
8.3)
TOC:
<
1.0
mg/
L
Conductivity:
321
(
315­
330)
µ
mhos/
cm
Stock
and
test
solution
preparation:
Primary
stock
prepared
in
dilution
water
at
88.4
mg/
L
and
mixed
until
all
test
substance
dissolved
prior
to
use.
After
mixing,
the
primary
stock
solution
was
proportionally
diluted
with
dilution
water
to
prepare
five
additional
stock
solutions
at
concentrations
of
44.2,
22.1,
11.0,
5.52,
and
2.76
mg/
L.
Stock
solutions
were
prepared
every
three
to
four
days
during
the
test.
The
six
stocks
were
injected
into
the
diluter
mixing
chambers
(
at
a
rate
of
6.0
mL/
minute)
where
they
were
mixed
with
dilution
water
(
at
a
rate
of
116
mL/
minute)
to
achieve
the
desired
test
concentrations.
Flow
through
rate:
Approximately
six
volume
additions
of
test
water
every
24­
hours
Stability
of
the
test
chemical
solutions:
Extremely
stable
Exposure
vessels:
9L
glass
aquaria
filled
with
approximately
7
L
of
test
solution
with
a
depth
of
approximately
17
cm.
Embryo
incubation
cups
were
constructed
from
glass
cylinders
approximately
50
mm
in
diameter
with
425
µ
m
nylon
screen
mesh
attached
to
the
bottom
with
ENV/
JM/
RD(
2002)
17/
FINAL
115
silicone
sealant.
The
cups
were
suspended
in
the
water
column
of
each
9L
glass
aquarium
and
attached
to
a
rocker
arm
with
a
reciprocating
motion
of
approximately
2
rpm.
Number
of
replicates:
four
Number
of
fish
per
replicate:
twenty
Number
of
concentrations:
six
plus
a
negative
control
Feeding:
Live
brine
shrimp
nauplii.
Fed
3
times
per
day
during
the
first
7
days
post­
hatch.
On
days
8
through
40
post­
hatch,
fed
3
times
daily
on
weekdays
and
2
times
daily
on
weekends.
Not
fed
for
at
least
48
hours
prior
to
the
termination
of
test
to
allow
for
gut
clearance
prior
to
weight
measurements.
Water
chemistry
during
the
study:
Dissolved
oxygen
range
(
0
 
47
days):
7.6
 
8.2
mg/
L
(
control
exposure)
7.6
 
8.2
mg/
L
(
1.2
mg/
L
exposure)
pH
range
(
0
 
47
days):
8.0
 
8.4
(
control
exposure)
8.0
 
8.4
(
1.2
mg/
L
exposure)
Test
temperature
range
(
0
 
47
days):
24.4
 
24.7oC
(
control
exposure)
24.3
 
24.7oC
(
1.2
mg/
L
exposure)
Method
of
calculating
mean
measured
concentrations:
arithmetic
mean
RESULTS
Nominal
concentrations:
Negative
control,
0.14,
0.29,
0.57,
1.1,
2.3,
4.6
mg/
L
Measured
concentrations:
<
LOQ,
0.15,
0.30,
0.60,
1.2,
2.4,
4.6
mg/
L
Element
value:
5­
day
hatchability
NOEC
=
4.6
mg/
L
42­
day
post­
hatch
survival
NOEC
=
0.30
mg/
L
42­
day
post­
hatch
growth
NOEC
=
0.30
mg/
L
42­
day
post­
hatch
survival
LOEC
=
0.60
mg/
L
All
element
values
based
on
mean
measured
concentrations
Statistical
evaluation
of
mortality:
The
statistical
difference
for
growth
at
concentrations
equal
to
and
higher
than
0.60
mg/
L
was
not
evaluated
due
to
a
significant
effect
on
survival.
No
statistically
significant
difference
between
the
negative
control
and
the
highest
concentration
tested
was
seen
for
hatchability.

Analytical
methodology:
Analyses
of
test
solutions
were
performed
at
Wildlife
International
Ltd.
using
high
performance
liquid
chromatography
with
mass
spectrometric
detection
(
HPLC/
MS).
When
determining
the
concentration
of
the
test
substance
in
the
test
solutions,
the
same
and
most
prominent
peak
response
for
perfluorooctane
sulfonate
was
used.
No
attempt
was
made
to
quantify
on
the
basis
of
individual
isomeric
components.
The
LOQ
(
limit
of
quantitation)
was
0.0458
mg/
L
in
this
study.
The
mean
percent
recovery
of
matrix
fortifications
analyzed
concurrently
during
sample
analysis
was
102.
Samples
collected
at
pre­
test
ranged
from
91.4
to
105%
of
nominal.
Samples
at
test
initiation
had
measured
values
from
95.5
to
114%
of
nominal.
Measured
values
for
samples
taken
at
test
termination
ranged
from
95.2
to
111%
of
nominal.
ENV/
JM/
RD(
2002)
17/
FINAL
116
Summary
of
analytical
chemistry
data:

Nominal
Test
Concentration,
mg/
L
Measured
duplicate
values
at
0,
4,
7,
14,
21,
28,
35,
42,
and
47
Days
respectively,
mg/
L
Mean
Measured
Concentration,
mg/
L
Percent
of
Nominal
Negative
Control
All
<
LOQ
­
­
0.14
0.147,
0.160,
0.141,
0.140,
0.144,
0.148,
0.134,
0.135,
0.153,
0.143,
0.160,
0.158,
0.179,
0.173,
0.157,
0.160,
0.147,
0.155
0.15
107
0.29
0.287,
0.277,
0.270,
0.289,
0.292,
0.296,
0.269,
0.266,
0.307,
0.315,
0.343,
0.341,
0.311,
0.325,
0.319,
0.313,
0.296,
0.276
0.30
103
0.57
0.571,
0.576,
0.619,
0.659,
0.597,
0.642,
0.539,
0.535,
0.608,
0.580,
0.639,
0.617,
0.646,
0.644,
0.575,
0.576,
0.545,
0.543
0.60
105
1.1
1.14,
1.13,
1.21,
1.25,
1.13,
1.23,
1.03,
1.10,
1.19,
1.24,
1.30,
1.31,
1.30,
1.31,
1.14,
1.19,
1.13,
1.09
1.2
109
2.3
2.21,
2.27,
2.52,
2.46,
2.43,
2.38,
fish
all
dead
at
Day
7
2.4
104
4.6
4.56,
4.40,
4.79,
4.79,
4.46,
4.76,
fish
all
dead
at
Day
7
4.6
100
Biological
Observations
Hatching
success
and
time
to
hatch:
All
viable
fathead
minnow
embryos
hatched
on
Day
4
or
5.
There
were
no
apparent
differences
between
the
time
to
hatch
in
the
negative
control
and
the
PFOS
treatment
groups.
Survival:
All
fish
surviving
to
test
termination
appeared
normal
with
no
overt
signs
of
sublethal
toxicity.
Fish
which
did
not
survive
generally
appeared
to
be
swimming
erratically
prior
to
death.
Growth:
Fish
exposed
to
PFOS
at
concentrations
of
0.15
or
0.30
mg/
L
for
42
days
post­
hatch
showed
no
statistically
significant
reduction
in
total
length,
wet
weight
or
dry
weight
in
comparison
to
the
negative
control.

Hatchability
Mean
Measured
Concentration
mg/
L
Number
of
Eggs
Exposed
Number
Hatched,
Day
3
Number
Hatched,
Day
4
Number
Hatched,
Day
5
Total
Number
Hatched
Percent
Hatching
Success
Negative
Control
80
0
20
54
74
93
0.15
80
0
18
58
76
95
0.3
80
0
14
58
72
90
0.6
80
0
28
48
76
95
1.2
80
0
25
49
74
93
2.4
80
0
16
59
75
94
4.6
80
0
14
60
74
93
ENV/
JM/
RD(
2002)
17/
FINAL
117
Larval
Survival
Mean
Measured
Concentration,
mg/
L
Percent
Survival,
Day
42
Negative
Control
88
0.15
79
0.3
81
0.6
66
1.2
5.4
2.4
0
4.6
0
Growth
Mean
Measured
Concentration,
mg/
L
Number
of
Surviving
Larvae
Total
Length
Mean
+
SD,
mm
Wet
Weight
Mean
+
SD,
mg
Dry
Weight
Mean
+
SD,
mg
Negative
Control
65
26.5
+
0.721
158
+
9.10
32.5
+
1.20
0.15
60
26.6
+
0.208
160
+
3.10
33.3
+
0.900
0.30
58
26.6
+
0.813
167
+
11.9
34.2
+
2.70
0.60
50
26.5
+
0.399
166
+
11.3
33.5
+
2.70
1.2
4
26.7
+
2.02
185
+
33.8
35.4
+
6.66
2.4
0
­
­
­
4.6
0
­
­
­

CONCLUSIONS
Fathead
minnows
exposed
to
potassium
perfluorooctanesulfonate
at
concentrations
<
0.30
mg/
L
for
42
days
post­
hatch
showed
no
statistically
significant
reductions
in
time
to
hatch,
hatching
success,
survival
or
growth.
The
most
sensitive
endpoint
in
this
study
was
post­
hatch
survival.

Submitter:
3M
Company,
Environmental
Laboratory
P.
O.
Box
33331
St.
Paul,
MN
55133,
USA.

DATA
QUALITY
Reliability:
Klimisch
ranking
=
1
REFERENCES
This
study
was
conducted
at
Wildlife
International
Ltd.,
Easton,
MD
at
the
request
of
the
3M
Company.

OTHER
Last
changed:
5/
3/
00
ENV/
JM/
RD(
2002)
17/
FINAL
118
Robust
Study
Report
Reference
No.
9
­
Semi­
Static
Life­
Cycle
Toxicity
Test
with
the
Cladoceran
(













)

TEST
SUBSTANCE
Identity:
Potassium
perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
The
test
substance
is
a
white
powder.
Sample
was
taken
from
3M
lot
number
217.
Sample
was
stored
under
ambient
conditions
prior
to
testing.
Purity
determined
to
be
90.49%
by
LC/
MS,
1H­
HMR,
19FNMR
and
elemental
analyses
techniques.

METHOD
Method:
OPPTS
850.1300,
OECD
Guideline
211,
and
ASTM
Standard
E
1193­
87.
Type:
Semi­
Static
Life­
Cycle
Toxicity
GLP:
Yes
Year
completed:
Study
completed
1999.
Report
completed
2000
Species:
Daphnia
magna
Supplier:
In­
house
cultures,
Wildlife
International,
Ltd.,
Easton,
MD,
USA
Analytical
monitoring:
PFOS
measured
on
days
0,
2,
11,
14,
18,
and
21.
Exposure
period:
21
days
Statistical
methods:
Survival
data
was
evaluated
on
first­
generation
daphnids,
the
number
of
live
young
and
the
length
and
dry
weight
of
the
surviving
first­
generation
daphnids.
Survival
data
were
analyzed
using
Fisher's
exact
test.
Reproduction
and
growth
(
length
and
dry
weight)
data
were
evaluated
for
normality
using
Shapiro­
Wilk's
test
and
for
homogeneity
of
variance
using
Bartlett's
test.
Analysis
of
variance
and
Dunnett's
test
was
used
to
identify
treatment
groups
that
were
statistically
significant
in
comparison
to
the
negative
control
(
p
<
0.05).
All
statistical
tests
were
performed
using
a
personal
computer
with
SPSS/
PC
Version
2.0
or
"
TOXSTAT
Release
3.5"
statistical
software.
Test
organism
age:
<
24­
hours
old
at
test
initiation
Pretreatment:
None
Test
Conditions
Dilution
water:
0.45
µ
m
filtered
well
water
passed
through
a
UV
sterilizer
to
remove
microorganisms
and
fine
particles
Dilution
water
chemistry
(
during
the
4­
week
period
immediately
preceding
the
test):
Hardness:
124
(
120­
128)
mg/
L
as
CaCO3
Alkalinity:
169
(
164­
172)
mg/
L
as
CaCO3
pH:
8.2
(
8.0­
8.3)
TOC:
<
1.0
mg/
L
Conductivity:
329
(
315­
340)
µ
mhos/
cm
Ca/
Mg
ratio:
35/
13.5
Na/
K
ratio:
21.3/
6.62
Stock
and
test
solution
preparation:
Primary
stock
solution
was
prepared
in
dilution
water
at
46
mg/
L.
It
was
stirred
until
all
test
substance
was
dissolved
prior
to
use.
After
mixing,
the
primary
stock
solution
was
proportionally
diluted
with
UV
sterilized
dilution
water
to
prepare
five
additional
stock
solutions
at
nominal
concentrations
of
1.4,
2.9,
5.7,
11,
and
23
mg/
L.
All
test
solutions
appeared
clear
and
colorless.
Renewal
rate:
Every
Monday,
Wednesday
and
Friday.
Exposure
vessels:
250­
mL
plastic
beakers
containing
approximately
200
mL
test
solution.
The
depth
was
approximately
5
cm.
Number
of
replicates:
10
ENV/
JM/
RD(
2002)
17/
FINAL
119
Number
of
test
organisms
per
replicate:
1
Number
of
concentrations:
6
plus
a
negative
control
Feeding:
Each
test
chamber
was
fed
0.3
mL
of
YCT
(
a
mixture
of
yeast,
Cerophyll
®
,
and
trout
chow
at
1800
mg
TSS/
L)
and
0.60
mL
of
Selenastrum
capricornutum
(
3.5
x
107
cells/
mL)
once
daily.
Lighting:
Colortone
®
50
fluorescent
lights.
Intensity
ranged
from
329
­
383
lux
at
the
water
surface.
Photoperiod
of
16­
hours
light,
8­
hours
dark
with
a
30­
minute
transition
period.
Water
chemistry
of
new
and
old
solutions
during
the
study:
Dissolved
oxygen
range
(
0
 
21
days):
8.3
 
8.9
mg/
L
(
negative
control
exposure)
8.3
 
9.0
mg/
L
(
12
mg/
L
exposure)
8.4
 
8.9
mg/
L*
(
48
mg/
L
exposure)
pH
range
(
0
 
21
days):
8.1
 
8.4
(
negative
control
exposure)
8.2
 
8.5
(
12
mg/
L
exposure)
8.4
 
8.5*
(
48
mg/
L
exposure)
Test
temperature
range
(
0
 
21
days):
19.4
 
20.1oC
(
negative
control
exposure)
19.4
 
20.1oC
(
12
mg/
L
exposure)
19.4
 
19.5
oC*
(
48
mg/
L
exposure)

*
(
Measurements
discontinued
at
Day
3
due
to
100%
mortality.

Element
basis:
Survival,
reproduction
and
growth.
Effect
concentrations
based
on
survival.
Method
of
calculating
mean
measured
concentrations:
arithmetic
mean
RESULTS
Nominal
concentrations:
Negative
control,
1.4,
2.9,
5.7,
11,
23,
46
mg/
L
Measured
concentrations:
<
LOQ,
1.5,
2.9,
5.6,
12,
24,
48
mg/
L
Element
value:
21­
day
NOEC
=
12
mg/
L
21­
day
LOEC
=
24
mg/
L
21­
day
MATC
=
17
mg/
L
2nd
generation
acute
survival
NOEC
=
12
mg/
L
All
element
values
based
on
mean
measured
concentrations.

Analytical
methodology:
Analyses
of
test
solutions
were
performed
at
Wildlife
International
Ltd.
using
high
performance
liquid
chromatography
with
mass
spectrometric
detection
(
HPLC/
MS).
When
determining
the
concentration
of
the
test
substance
in
the
test
solutions,
the
same
and
most
prominent
peak
response
for
the
test
substance
was
used.
No
attempt
was
made
to
quantify
on
the
basis
of
individual
isomeric
components.
The
LOQ
(
limit
of
quantitation)
was
0.458
mg/
L
in
this
study.
The
mean
procedural
recovery
of
matrix
fortifications
analyzed
concurrently
during
sample
analysis
was
104%.
Measured
values
of
new
samples
ranged
from
94
to
121%
of
nominal.
Measured
values
from
the
old
solutions
ranged
from
90
to
108%
of
nominal
values.
PFOS
was
stable
throughout
the
renewal
periods.
ENV/
JM/
RD(
2002)
17/
FINAL
120
Summary
of
analytical
chemistry
data:

Nominal
Test
Concentration
mg/
L
Measured
duplicate
values
at
0,
2,
11,
14,
18,
and
21
Days
respectively,
mg/
L
Mean
Measured
Concentration
mg/
L
Percent
of
Nominal
Negative
Control
All
<
LOQ
­
­
1.4
1.78,
1.72,
1.58,
1.56,
1.38,
1.47,
1.36,
1.32,
1.38,
1.43,
1.50,
1.45
1.5
107
2.9
3.20,
3.05,
3.01,
3.07,
2.75,
2.77,
2.85,
2.71,
2.79,
2.81,
2.81,
2.82
2.9
100
5.7
5.97,
5.87,
5.65,
5.72,
5.63,
5.59,
5.36,
5.39,
5.58,
5.75,
5.24,
5.37
5.6
98
11
11.5,
11.5,
11.6,
11.8,
11.3,
11.3,
11.2,
11.6,
11.8,
11.6,
11.5,
11.3
12
109
23
24.2,
23.1,
24.0,
24.6,
22.8,
22.5,
23.6,
23.1,
24.8,
25.0,
all
daphnids
dead
after
18­
days
exposure
24
104
46
47.3,
48.0,
49.1,
49.4,
all
daphnids
dead
after
2­
days
exposure
48
104
NOTE:
Mean
measured
concentrations
were
determined
from
new
(
renewal
solutions)
and
corresponding
old
solutions
during
each
week
of
the
test.
Days
0,
11,
and
18
are
"
new"
and
days
2,
24,
and
21
are
"
old".

Biological
Observations
Survival:
All
surviving
first
generation
daphnids
appeared
normal
at
test
termination.
Survival
in
the
24
and
48
mg/
L
treatments
was
statistically
significantly
different
from
the
negative
control
group.
Reproduction:
Daphnids
in
the
control
and
treatment
groups
<
12
mg/
L
started
producing
neonates
on
Day
9.
The
Bonferroni
t­
test
showed
that
reproduction
was
not
significantly
reduced
in
any
treatment
group
<
12
mg/
L
(
p
>
0.05).
The
24
and
48
mg/
L
treatment
groups
were
not
included
in
the
statistical
analysis
of
the
reproduction
data
due
to
a
statistically
significant
effect
on
survival.
Growth:
The
Bonferroni
t­
test
showed
that
mean
length
and
dry
weight
in
the
treatment
groups
<
12
mg/
L
were
not
significantly
reduced
in
comparison
to
the
negative
control
(
p
>
0.05).
Second
Generation
Acute
Exposure:
After
48­
hours
of
exposure,
survival
in
the
negative
control
was
95%.
Survival
in
the
1.5,
2.9,
5.6,
12,
and
24
mg/
L
treatment
groups
was
100,
100,
100,
90,
and
0%
respectively.
Survival
in
the
24
mg/
L
treatment
group
was
significantly
different
from
the
negative
control
(
p
<
0.05).

Summary
of
Percent
Mortality
Mean
Measured
Concentration,
mg/
L
Day
7
Day
14
Day
21
Negative
Control
0
0
0
1.5
0
0
10
2.9
0
0
10
5.6
0
0
10
12
0
10
10
24
70
90
100
48
100
100
100
ENV/
JM/
RD(
2002)
17/
FINAL
121
Second
Generation
Mortality
Mean
Measured
Concentrations,
mg/
L
Total
Number
Exposed
Number
Alive
after
48­
hours
Cumulative
Percent
Dead
Negative
Control
20
19
5
1.5
20
20
0
2.9
20
20
0
5.6
20
20
0
12
20
18
10
24
8
0
100
Summary
of
Length
and
Dry
Weight
of
Surviving
Individually­
Exposed
First­
Generation
Daphnids
Mean
Measured
Concentration,
mg/
L
Number
of
Surviving
Daphnids
Total
Length,
Mean
+
SD,
mm
Dry
Weight,
Mean
+
SD,
mg
Negative
Control
10
4.65
+
0.111
0.695
+
0.100
1.5
9
4.66+
0.118
0.669
+
0.0623
2.9
9
4.62
+
0.100
0.724
+
0.110
5.6
9
4.61
+
0.124
0.727
+
0.0665
12
9
4.59
+
0.102
0.723
+
0.0661
24
0
­
­
­
­
48
0
­
­
­
­

Reproduction
Mean
Measured
Concentration,
mg/
L
Number
of
Surviving
Daphnids
Mean
Live
Young/
Surviving
Adult
Daphnid
(+
SD)
First
Day
of
Reproduction
Total
Number
of
Dead
/
Immobile
Neonates
Total
Number
of
Aborted
Eggs
Negative
Control
10
122
+
19.2
9
0
0
1.5
9
142
+
24.7
9
0
0
2.9
9
136
+
17.9
9
0
0
5.6
9
132
+
19.5
9
0
0
12
9
119
+
26.5
9
1
0
24
0
­
­
11
10
0
48
0
­
­
None
­
­
­
­

CONCLUSIONS
There
were
no
adverse
effects
on
survival,
reproduction
or
growth
of
Daphnia
magna
exposed
to
the
test
substance
at
concentration
<
12
mg/
L
for
21
days.
Daphnia
magna
exposed
to
24
and
48
mg/
L
had
significantly
reduced
survival.

Author
and/
or
submitter:
3M
Company,
Environmental
Laboratory
P.
O.
Box
33331
St.
Paul,
MN
55133,
USA.

DATA
QUALITY
Reliability:
Klimisch
ranking
1.
ENV/
JM/
RD(
2002)
17/
FINAL
122
REFERENCES
This
study
was
conducted
at
Wildlife
International,
Ltd.
Easton,
MD
at
the
request
of
the
3M
Company.

OTHER
Last
changed:
5/
3/
00
ENV/
JM/
RD(
2002)
17/
FINAL
123
Robust
Study
Report
Reference
No.
10
­
Flow­
through
Life­
Cycle
Toxicity
Test
with
the
Saltwater
Mysid
(
















)

TEST
SUBSTANCE
Identity:
Potassium
perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
The
test
substance
is
a
white
powder.
Sample
was
taken
from
3M
lot
number
217.
Sample
was
stored
under
ambient
conditions
prior
to
testing.
Purity
determined
to
be
90.49%
by
LC/
MS,
1H­
HMR,
19FNMR
and
elemental
analyses
techniques.

METHOD
Method:
OPPTS
850.1350
Type:
Flow­
through
chronic
GLP:
Yes
Year
completed:
Study
completed
1999.
Report
completed
2000
Species:
Mysidopsis
bahia
Supplier:
In­
house
cultures,
Wildlife
International,
Ltd.,
Easton,
MD,
USA
Analytical
monitoring:
PFOS
measured
on
days
0,
7,
14,
21,
28,
and
35
Exposure
period:
35
days
Statistical
methods:
Survival
data
was
evaluated
(
prior
to
pairing
and
after
pairing)
using
2
x
2
contingency
tables
to
identify
treatment
groups
that
showed
a
statistically
significant
difference
(
p<
0.05)
from
the
negative
control
group.
All
continuous­
variable
data
(
reproduction
and
growth)
were
evaluated
for
normality
using
Shapiro­
Wilk's
test
and
for
homogeneity
of
variance
using
Bartlett's
test.
Analysis
of
variance
and
Dunnett's
test
were
used
to
evaluate
differences
between
treatment
and
control
means.
All
statistical
tests
were
performed
using
a
personal
computer
with
SPSS/
PC
Version
2.0
or
"
TOXSTAT
Release
3.5"
statistical
software.
Test
mysids
age:
<
24­
hours
old
at
test
initiation
Pretreatment:
None
Test
Conditions:
Natural
seawater
diluted
to
20%
o
with
well
water,
0.45
µ
m
filtered.
Dilution
water
chemistry
(
during
the
4­
week
period
immediately
preceding
the
test):
Salinity:
20
(
20­
20)
%
o
TOC:
<
1.0
mg/
L
Stock
and
test
solution
preparation:
Primary
stock
prepared
at
0.0895
mg/
L
and
mixed
for
approximately
24
hours
prior
to
use.
After
mixing,
the
primary
stock
solution
was
proportionally
diluted
with
dilution
water
to
prepare
five
additional
stock
solutions
at
concentrations
of
0.0447,
0.0224,
0.0112,
0.00559,
and
0.00280
mg/
L.
The
six
stocks
were
injected
into
the
diluter
mixing
chambers
(
at
a
rate
of
4.60
mL/
minute)
where
they
were
mixed
with
dilution
water
(
at
a
rate
of
150
mL/
minute)
to
achieve
the
desired
test
concentrations.
Flow
through
rate:
Approximately
eleven
volume
additions
of
test
water
every
24
hours
Stability
of
the
test
chemical
solutions:
Extremely
stable
Exposure
vessels:
Prior
to
pairing,
mysids
placed
in
glass
beakers
with
nylon
mesh
screen
attached
to
two
holes
on
opposite
sides.
After
reaching
sexual
maturity,
pairs
placed
in
glass
petri
dishes
with
sides
of
nylon
mesh
screen
attached
with
silicone
adhesive.
Both
pre­
pairing
and
postpairing
exposure
vessels
were
placed
in
9L
glass
aquaria
filled
with
approximately
5
L
of
test
solution.
The
depth
was
approximately
6.2
cm
prior
to
pairing
and
5.5
cm
after
pairing.
ENV/
JM/
RD(
2002)
17/
FINAL
124
The
test
chambers
for
the
second
generation
exposure
were
2L
beakers
with
1L
of
test
solution
which
was
dipped
out
of
a
test
chamber
from
the
appropriate
treatment
group.
Number
of
replicates:
four
Number
of
concentrations:
six
plus
a
negative
control
Number
of
fish
per
replicate:
Fifteen
juveniles
before
pairing,
5
pairs
(
10
adults)
when
possible
after
pairing.
Feeding:
Fed
live
brine
shrimp
nauplii
3
or
four
times
per
day.
Not
fed
the
last
day
of
the
test.
Water
chemistry
during
the
study:
Dissolved
oxygen
range
(
0
 
35
days):
6.0
 
6.4
mg/
L
(
control
exposure)
5.9
 
6.3
mg/
L
(
1.3
mg/
L
exposure)
pH
range
(
0
 
35
days):
8.2
 
8.4
(
control
exposure)
8.3
 
8.4
(
1.3
mg/
L
exposure)
Test
temperature
range
(
0
 
35
days):
24.5
 
25.2oC
(
control
exposure)
24.4
 
25.1oC
(
1.3
mg/
L
exposure)

Method
of
calculating
mean
measured
concentrations:
arithmetic
mean
RESULTS
Nominal
concentrations:
Negative
control,
0.086,
0.17,
0.34,
0.69,
1.4,
2.7
mg/
L
Measured
concentrations:
<
LOQ,
0.057,
0.12,
0.25,
0.55,
1.3,
2.6
mg/
L
Element
value:
20­
day
survival
(
pre­
pairing)
NOEC
=
0.55
mg/
L
35­
day
(
post­
pairing)
survival
NOEC
=
0.55
mg/
L
35­
day
reproduction
NOEC
=
0.25
mg/
L
35­
day
growth
NOEC
=
0.25
mg/
L
35­
day
reprod
&
growth
LOEC
=
0.55
mg/
L
2nd
generation
acute
survival
NOEC
=
0.55
mg/
L
(
highest
concentration
tested)

All
element
values
based
on
mean
measured
concentrations
Analytical
methodology:
Analyses
of
test
solutions
were
performed
at
Wildlife
International
Ltd.
using
high
performance
liquid
chromatography
with
mass
spectrometric
detection
(
HPLC/
MS).
When
determining
the
concentration
of
the
test
substance
in
the
test
solutions,
the
same
and
most
prominent
peak
response
for
perfluorooctanesulfonate
was
used.
No
attempt
was
made
to
quantify
on
the
basis
of
individual
isomeric
components.
The
LOQ
(
limit
of
quantitation)
was
0.0458
mg/
L
in
this
study.
The
mean
percent
recovery
of
matrix
fortifications
analyzed
concurrently
during
sample
analysis
was
92.8.
Samples
collected
at
pretest
ranged
from
57.4
to
99.3%
of
nominal.
Samples
at
test
initiation
had
measured
values
from
67.1
to
103%
of
nominal.
Measured
values
for
samples
taken
at
test
termination
ranged
from
59.8
to
90.0%
of
nominal.
ENV/
JM/
RD(
2002)
17/
FINAL
125
Summary
of
analytical
chemistry
data:

Nominal
Test
Concentration,
mg/
L
Measured
duplicate
values
at
0,
7,
14,
21,
28,
35,
Days
respectively,
mg/
L
Mean
Measured
Concentration,
mg/
L
Percent
of
Nominal
Negative
Control
All
<
LOQ
­
­
0.086
0.0694,
0.0578,
0.0478,
0.0619,
0.0606,
0.0614,
0.0554,
0.0509,
0.0515,
0.0569,
0.0580,
0.0514
0.057
66
0.17
0.125,
0.114,
0.0778,
0.125,
0.124,
0.127,
0.0970,
0.112,
0.122,
0.128,
0.124,
0.119
0.12
71
0.34
0.289,
0.286,
0.231,
0.197,
0.276,
0.253,
0.227,
0.212,
0.262,
0.271,
0.278,
0.251
0.25
74
0.69
0.562,
0.659,
0.581,
0.450,
0.543,
0.542,
0.516,
0.528,
0.529,
0.544,
0.556,
0.583
0.55
80
1.4
1.23,
1.32,
1.13,
1.20,
1.35,
1.27,
1.23,
1.15,
1.39,
1.39,
1.26,
1.20
1.3
93
2.7
2.56,
2.79,
2.58,
2.30,
2.54,
2.69,
all
mysids
dead
after
14­
days
exposure
2.6
96
Biological
observations
Survival:
All
surviving
mysids
appeared
normal.
Survival
in
the
1.3
and
2.6
mg/
L
treatments
were
statistically
significantly
different
from
the
negative
control
group.
Reproduction:
The
day
of
first
brood
release
in
this
study
was
Day
22.
Dunnett's
test
showed
that
reproduction
was
significantly
reduced
in
the
0.55
mg/
L
treatment
group
when
compared
to
the
negative
control
(
p
<
0.05).
The
1.3
and
2.6
mg/
L
treatment
groups
were
not
included
in
the
statistical
analysis
of
the
reproduction
data
due
to
a
statistically
significant
difference
in
survival.
Growth:
Mysids
exposed
to
PFOS
at
concentrations
<
0.25
mg/
L
showed
no
statistically
significant
reductions
in
length
or
dry
weight
(
p
<
0.05).
Second
Generation
Acute
Exposure:
Survival
in
all
PFOS
treatment
groups
was
>
95%
and
was
not
statistically
different
from
the
controls.
All
surviving
mysids
in
the
second­
generation
exposure
appeared
normal
with
no
overt
signs
of
toxicity.

Percent
Survival
Mean
Measured
Concentration,
mg/
L
Juvenile
Pre­
Pairing
Survival,
Day
20
Adult
Post­
Pairing
Survival,
Day35
Negative
Control
78
92
0.057
92
96
0.12
75
90
0.25
82
97
0.55
83
95
1.3
32
57
2.6
0
­
ENV/
JM/
RD(
2002)
17/
FINAL
126
Second
Generation
Survival
Mean
Measured
Concentrations,
mg/
L
Total
Number
Exposed
Number
Alive
after
96­
hours
Percent
Survival
Negative
Control
71
68
96
0.057
65
63
97
0.12
83
79
95
0.25
62
59
95
0.55
13
13
100
Adult
Mysid
Growth
Mean
Measured
Concentration,
mg/
L
Number
of
Surviving
Mysids/
Number
Exposed
Total
Length,
Mean
+
SD,
mm
Dry
Weight,
Mean
+
SD,
mg
Negative
Control
36/
39
6.43
+
0.0634
0.634
+
0.0510
0.057
44/
46
6.43
+
0.0729
0.599
+
0.0276
0.12
36/
40
6.56
+
0.105
0.641
+
0.0241
0.25
36/
37
6.40
+
0.0548
0.622
+
0.0227
0.55
35/
37
6.14
+
0.0794
0.562
+
0.00624
1.3
8/
14
5.85
+
0.178
0.436
+
0.0441
Reproduction
Mean
Measured
Concentration,
mg/
L
Replicate
Number
of
Reproductive
Days
Number
of
Young
Mean
Number
of
Young/
Reproductive
Day
Overall
Mean+
SD
Negative
control
A
70
18
0.257
0.315
+
0.0925
B
53
14
0.264
C
70
20
0.286
D
42
19
0.452
0.57
A
60
17
0.283
0.261
+
0.0873
B
70
14
0.200
C
70
13
0.186
D
56
21
0.375
0.12
A
70
21
0.300
0.361
+
0.101
B
46
22
0.478
C
54
22
0.407
D
70
18
0.257
0.25
A
70
19
0.271
0.252
+
0.0723
B
56
12
0.214
C
61
21
0.344
D
56
10
0.179
0.55
A
54
3
0.0556
0.0559
+
0.0376
B
56
6
0.107
C
70
3
0.0429
D
56
1
0.0179
ENV/
JM/
RD(
2002)
17/
FINAL
127
1.3
A
22
0
­
­
B
14
0
­
C
0
0
­
D
11
0
­

CONCLUSIONS
There
were
no
statistically
significant
effects
on
survival,
reproduction
or
growth
of
mysid
shrimp
exposed
to
potassium
perfluorooctanesulfonate
at
concentrations
<
0.25
mg/
L
for
35
days.
Reproduction,
length
and
dry
weight
were
the
most
sensitive
biological
endpoints
in
this
study.
Second
generation
mysids
exposed
to
PFOS
during
a
static
96­
hour
exposure
showed
no
adverse
effects.

Submitter:
3M
Company,
Environmental
Laboratory
P.
O.
Box
33331
St.
Paul,
MN
55133,
USA
DATA
QUALITY
Reliability:
Klimisch
ranking
=
1
REFERENCES
This
study
was
conducted
at
Wildlife
International
Ltd.,
Easton,
MD
at
the
request
of
the
3M
Company.

OTHER
Last
changed:
7/
9/
01
ENV/
JM/
RD(
2002)
17/
FINAL
128
Robust
Study
Report
Reference
No.
11
 
Dietary
LC50
Study
with
the
Mallard
duck
TEST
SUBSTANCE
Identity:
Potassium
perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
The
test
substance
is
a
white
powder.
Sample
was
taken
from
3M
lot
number
217.
Sample
was
stored
under
ambient
conditions
prior
to
testing.
Purity
determined
to
be
90.49%
by
LC/
MS,
'
H­
HMR,
19FNMR
and
elemental
analyses
techniques.

METHOD
Method:
OPPTS
850.2200,
OECD
206,
and
FIFRA
Subdivision
E.
Section
71­
2
Type:
Dietary
acute
GLP:
Yes
Year
completed:
2000
Species:
Anas
platyrhynchos
Supplier:
Whistling
Wings,
Inc.,
Hanover,
IL,
USA
Analytical
monitoring:
PFOS
measured
on
Day
0
for
homogeneity
in
feed
and
verification,
and
Day
5
for
stability.
Test
phases:
Acclimation
­
9
days
Exposure
­
5
days
Post­
exposure
observation
­
3
or
17
days
Statistical
methods:
LC50
values
calculated
by
probit
analysis
using
the
computer
software
of
C.
E.
Stephan.
Body
weight
data
were
compared
by
Dunnett's
test
using
TOXSTAT
software.
No
statistical
analyses
were
applied
to
feed
consumption
data.
Test
bird
age:
10
days
Pretreatment:
None
Test
conditions:
Housing
and
environmental
conditions:
Indoors
In
batteries
of
thermostatically
controlled
brooding
pens.
Floor
space
of
each
pen
measured
approximately
62
x
90
cm.
Ceiling
height
was
approximately
25.5
cm.
External
walls,
ceilings
and
floors
were
constructed
of
galvanized
steel
wire
and
sheeting.
ldentification:
Each
group
of
birds
Identified
by
pen
number
and
test
concentration.
Individuals
identified
by
wing
bands.
Number
of
replicates:
Six
for
controls,
two
for
each
treatment
group
Number
of
ducks
per
replicate:
five
Number
of
concentrations:
Eight
plus
a
negative
control
Feed
and
water:
Game
bird
ration
formulated
as
below,
water
from
the
town
of
Easton
public
water
supply.
Both
provided
ad
libitum
during
acclimation
and
testing.
ENV/
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2002)
17/
FINAL
129
Game
Bird
Ration
Ingredients
Percent
Fine
Corn
Meal
44.83
Soy
Bean
Meal,
48%
Protein
30.65
Wheat
Midds
6.50
Protein
Base
6.00
Agway
Special,
60%
Protein
4.00
Alfalfa
Meal,
20%
Protein
3.00
Dried
Whey
2.50
Ground
Limestone
0.90
Eastman
CalPhos
0.60
Methionine
Premix
+
Liquid
0.35
Vitamin
and
Mineral
Premix
(
see
below)
0.32
GL
Form
(
Fermatco)
1
0.25
Salt
iodized
0.10
1Fermentation
by­
products
(
source
of
unidentified
growth
factors)

Vitamin
and
Mineral
Premix
Vitamin
or
Mineral
Amount
Per
Ton
Vitamin
D3
2,000,000
I.
C.
U.
Vitamin
A
7,000,000
I.
U.
Riboflavin
6
g
Niacin
40
g
Pantothenic
Acid
10
g
Vitamin
B12
8
mg
Folic
Acid
600
mg
Biotin
64
mg
Pyridoxine
1.2
g
Thiamine
1.2
g
Vitamin
E
20,000
I.
U.
Vitamin
K
(
Menadione
dimethylpyrimidinol
bisulfite)
5.8
g
Manganese
102
g
Zinc
47
g
Copper
6.8
g
Iodine
1.5
g
Iron
51
g
Selenium
182
g
Prophylaxis:
None
Brooding
compartment
mean
temperature:
38
±
2oC
Ambient
room
mean
temperature:
25.2
±
0.7oC
Average
relative
humidity:
53
±
18%
Photoperiod:
Sixteen
hours
light
per
day
Lighting:
fluorescent
lights
which
closely
approximate
noon­
day
sunlight;
average
approximately
207
lux.
ENV/
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2002)
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130
Test
diet
preparation:
Test
substance
mixed
directly
into
the
ration
by
means
of
a
Hobart
mixer.
No
carrier
was
used.

Diet
sampling:
Homogeneity
of
the
test
substance
in
the
diet
evaluated
by
collecting
six
samples
from
the
9.1
ppm
and
six
from
the
lowest
and
highest
concentration.
Samples
collected
from
the
top,
middle,
and
bottom
of
the
left
and
right
sections
of
the
mixing
vessel.
These
samples
also
served
as
the
verification
samples
for
these
concentrations.
Two
verification
samples
from
the
remaining
concentrations
and
one
from
the
control
were
collected
at
preparation
on
Day
0.
Stability
samples
were
collected
at
the
end
of
the
exposure
period
(
Day
5)
from
the
control
(
one
sample)
and
each
treatment
group
(
two
samples
each).

RESULTS
Nominal
concentrations:
Negative
control,
9.1,
18.3,
36.6,
73.2,
146,
293,
586,
and
1171
ppm
Measured
concentrations:
<
LOQ,
9.8,
19.5,
40.2,
74.5,
174,
291,
537,
and
1196
ppm
Element
value:
Dietary
LC50
=
628
(
448
­
958)
ppm
No
mortality
concentration
146
ppm
NOEC
(
body
weight
gain)
36.6
ppm
All
element
values
based
on
nominal
concentrations
Analytical
Methodology:
Diet
samples
were
extracted
with
methanol.
Analyses
of
test
solutions
were
performed
at
Wildlife
International
Ltd.
using
high
performance
liquid
chromatography
with
mass
spectrometric
detection
(
HPLC/
MS).
When
determining
the
concentration
of
the
test
substance
in
the
test
solutions,
the
same
and
most
prominent
peak
response
for
perfluorooctanesulfonate
was
used.
No
attempt
was
made
to
quantify
on
the
basis
of
individual
isomeric
components.
The
LOQ
(
limit
of
quantitation)
was
1.15
ppm
in
this
study.
The
mean
percent
recovery
of
matrix
fortifications
analyzed
concurrently
during
sample
analysis
was
94.7.
Samples
collected
for
determination
of
homogeneity
in
diet
ranged
from
102
­
108%
of
nominal.
Samples
collected
for
verification
in
diet
had
measured
values
from
92
to
119%
of
nominal.
Measured
values
for
ambient
stability
samples
taken
at
Day
5
ranged
from
94
 
130%
of
nominal.

Summary
of
analytical
chemistry
data
Homogeneity
in
Avian
diet
Nominal
Test
Concentration,
ppm
Measured
Values
at
Day
0,
ppm
Mean
Measured
Concentration,
ppm
Percent
of
Nominal
9.1
9.52,
9.70,
9.79,
8.09,
10.9,
10.5
9.8
108
18.3
18.5,
23.4,
18.3,
17.3,
19.4,
19.9
19.6
107
1171
1239,
1221,
1118,
1301,
1163,
1133
1196
102
Verification
in
avian
diet
Nominal
Test
Concentration,
ppm
Measured
Duplicate
Concentrations
at
Day
0,
ppm
Mean
Measured
Concentration,
ppm
Percent
of
Nominal
Negative
Control
<
LOQ
­
­
36.6
46.7,
34.6
40.2
110
73.2
77.8,
71.2
74.5
102
146
176,
172
174
119
293­
274,
307
291
99
586
550,
523
537
92
ENV/
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2002)
17/
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131
Ambient
stability
in
avian
diet
Day
0
Mean
Measured
Concentration,
ppm
Measured
Duplicate
Concentrations
at
Day
5,
ppm
Mean
Measured
Concentration,
ppm
Mean
Percent
of
Day
0
Negative
Control
<
LOQ
­
9.8
12.3,
11.0
11.7
119
19.5
18.2,
19.7
19.0
97
40.2
47.9,
56.8
52.4
130
74.5
77.6,
77.9
77.8
104
174
167,
160
164
94.3
291
297,
293
295
101
537
552,
530
541
101
1196
1150,
1122
1136
95
Biological
observations
Survival
and
clinical
observations:
No
mortalities
occurred
in
the
control
group,
and
all
birds
were
normal
in
appearance
and
behavior
throughout
the
test.
The
first
deaths
occurred
on
day
4
in
the
1171
ppm
treatment.
Mortality
occurred
through
Day
8
in
all
dose
groups
>
293
ppm
with
some
of
the
deaths
being
during
the
post­
exposure
period.
There
were
no
treatment­
related
mortalities
or
overt
signs
of
toxicity
at
concentrations
<
146
ppm.
Birds
at
all
concentrations
>
293
ppm
displayed
signs
of
toxicity
including
reduced
reaction
to
stimuli
(
sound
and
motion),
loss
of
coordination,
ruffled
appearance,
lethargy
and
lower
limb
weakness.
Birds
at
the
1171
ppm
level
also
displayed
prostrate
posture,
depression
and
convulsions
through
Day
8.
Recovery
with
normal
appearance
and
behavior
was
noted
from
Day
9
through
test
termination
Body
weight
gain:
When
compared
to
the
control
group,
there
were
no
apparent
treatment
related
effects
on
body
weight
among
the
birds
in
concentrations
<
36.6
ppm.
During
the
Day
8­
15
and
Day
15­
22
post­
exposure
periods,
body
weight
gain
appeared
comparable
among
all
groups.
There
was
a
statistically
significant
(
p
<
0.05)
reduction
in
weight
gain
at
the
9.1
ppm
level
for
the
Day
0­
5
and
Day
5­
8
periods.
However,
differences
from
the
control
group
at
the
9.1
ppm
level
appear
to
be
due
to
a
lower
mean
Day
0
body
weight
for
the
9.1
ppm
level,
and
were
not
dose
responsive.
Therefore,
these
differences
were
not
considered
treatment
related.
Marked,
treatment­
related,
concentration
responsive
effects
on
body
weight
was
noted
in
concentrations
>
73.2
ppm
for
Days
0­
5;
Day
5­
8
post­
exposure
weight
gain
continued
to
be
reduced
at
concentrations
>
293
ppm.
Feed
Consumption:
When
compared
to
the
control
group,
there
was
a
marked
reduction
in
feed
consumption
in
the
treatment
groups
>
293
ppm
throughout
the
study.
Gross
Necropsy:
All
birds
that
died
during
the
study,
half
of
those
surviving
at
Day
8
and
the
rest
at
test
termination
were
subjected
to
a
gross
necropsy.
Necropsy
results
for
birds
found
dead
were
similar,
including
thin
condition,
loss
of
muscle
mass,
altered
spleen
color,
empty
crops,
and
empty
gastrointestinal
tracts.
These
necropsy
findings
were
considered
to
be
treatment
related.

Percent
Cumulative
Mortality
Nominal
Concentration,
ppm
Day
1
Day
2
Day
3
Day
4
Day
5
Day
6
Day
7
Day
8*

Negative
Control
0
0
0
0
0
0
0
0
9.1
0
0
0
0
0
0
0
0
18.3
0
0
0
0
0
0
0
0
36.6
o
0
0
0
0
0
0
0
ENV/
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2002)
17/
FINAL
132
73.2
0
0
0
0
0
0
0
0
146
0
0
0
0
0
0
0
0
293
0
0
0
0
0
0
10
20
586
0
0
0
0
10
20
30
30
1171
0
0
0
20
60
80
90
90
*
No
mortalities
occurred
in
any
of
the
treatment
levels
from
Day
8
to
Day
22
Bodyweight
(
grams)

Exposure
Period
Recovery
Period
Nominal
Concentration,
ppm
Mean
Body
Weight,
Day
0
Mean
Body
Weight
Change
Day
0­
5
Mean
Body
Weight
Change
Day
5­
8
Mean
Body
Weight
Change
Day
8­
15
Mean
Body
Weight
Change
Day
15­
22
Mean
Total
Body
Weight
Change
Day
8­
22
Mean
Body
Weight
Day
22
Negative
Control
135
144
101
230
183
413
823
9.1
119
108
91
230
198
427
773
18.3
146
131
100
241
186
427
811
36.6
147
128
100
243
208
451
828
73.2
143
117
82
216
203
418
782
146
143
100
89
232
124
356
688
293
129
32
57
256
234
490
701
586
144
­
6
36
221
219
439
613
1171
147
­
37
1531
198
251
449
634
Mean
average
feed
consumption
Nominal
Concentration,
ppm
Grams
food/
bird/
day
Days
0­
5
Grams
food/
bird/
day
Days
6­
8
Grams
food/
bird/
day
Days
8­
15
Grams
food/
bird/
day
Days
15­
22
Negative
Control
92
125
171
180
9.1
73
117
172
198
18.3
91
132
186
204
36.6
94
125
165
179
73.2
77
101
148
173
146
105
159
159
164
293
44
63
109
132
586
36
55
114
143
1171
22
25
106
154
Gross
pathological
observations
from
Birds
that
died
in
study
Finding
293ppm
N
=
2
586ppm
N
=
3
1171ppm
N
=
9
Crop
empty
0
2
7
Emaciated
1
1
4
G.
I.
Tract,
primarily
empty
1
1
1
Gizzard
contents
bile
stained
0
2
4
Gizzard,
empty
0
0
1
Intestinal
contents,
black
and
tar­
like
0
0
1
Keel
prominent
0
0
2
ENV/
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2002)
17/
FINAL
133
Kidneys,
pale
0
0
1
Loss
of
muscle
mass
2
2
5
Spleen,
grey
0
0
1
Spleen,
small
and
pale
1
1
2
Spleen,
pale
0
2
3
Thin
1
1
4
CONCLUSIONS
The
dietary
LC50
value
for
Mallard
Duck
exposed
to
PFOS
was
determined
to
be
628
ppm
with
a
95%
confidence
interval
of
448
to
958
ppm.
The
slope
of
the
concentration­
response
curve
was
3.67
and
the
chi­
square
value
was
2.13.
The
no
mortality
concentration
was
146
ppm.
Based
upon
reductions
in
body
weight
gain
at
the
73.2
ppm
test
concentration,
the
no
observed
effect
concentration
was
36.6
ppm.

Submitter:
3M
Company,
Environmental
Laboratory,
P.
O.
Box
33331,
St.
Paul,
Minnesota,
55133,
USA
DATA
QUALITY
Reliability:
Klimisch
ranking
1
REFERENCES
This
study
was
conducted
at
Wildlife
International
Ltd.,
Easton,
MD
at
the
request
of
the
3M
Company.

OTHER
Last
changed:
5/
1/
00
ENV/
JM/
RD(
2002)
17/
FINAL
134
Robust
Study
Report
Reference
No.
12
 
Dietary
LC50
Study
with
the
Northern
Bobwhite
quail
TEST
SUBSTANCE
Identity:
Potassium
perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks
field:
The
test
substance
is
a
white
powder.
Sample
was
taken
from
3M
lot
number
217.
Sample
was
stored
under
ambient
conditions
prior
to
testing.
Purity
determined
to
be
90.49%
by
LC/
MS,
1H­
HMR,
19F­
NMR
and
elemental
analyses
techniques.

METHOD
Method:
OPPTS
850.2200,
OECD
205,
and
FIFRA
Subdivision
E.
Section
71­
2
Type:
Dietary
acute
GLP:
Yes
Year
completed:
1999
(
Test),
2000
(
Report)
Species:
Colinus
virginianus
Supplier:
Wildlife
International
Ltd.
Production
Flock,
Easton,
Maryland,
USA
Analytical
monitoring:
Test
substance
concentration
in
standards
and
samples
were
determined
by
reversed­
phase
HPLC
and
mass
spectroscopy.
PFOS
measured
on
Day
0
for
homogeneity
in
feed
and
verification,
and
Day
5
for
stability.
Test
phases:
Acclimation
 
10
days
Exposure
 
5
days
Post­
exposure
observation
 
3
or
17
days
Statistical
methods:
LC50
values
calculated
by
probit
analysis
using
the
computer
software
of
C.
E.
Stephan.
Body
weight
data
were
compared
by
Dunnett's
test
using
TOXSTAT
software.
No
statistical
analyses
were
applied
to
feed
consumption
data.
Test
bird
age:
10
days
Pretreatment:
None
Test
conditions:
Housing
and
environmental
conditions:
Indoors
in
batteries
of
thermostatically
controlled
brooding
pens.
Each
pen's
floor
space
measured
approximately
72
x
90
cm.
Ceiling
height
was
approximately
23
cm.
External
walls,
ceilings
and
floors
were
constructed
of
galvanized
steel
wire
and
sheeting.
Identification:
Each
group
of
birds
Identified
by
pen
number
and
test
concentration.
Individuals
Identified
by
leg
bands.
Number
of
replicates:
Six
for
controls,
two
for
each
treatment
group
Number
of
bobwhite
per
replicate:
five
Number
of
concentrations:
seven
plus
a
negative
control
Feed
and
water:
Game
bird
ration
formulated
as
below,
water
from
the
town
of
Easton
public
water
supply.
Both
provided
ad
libitum
during
acclimation
and
testing.
ENV/
JM/
RD(
2002)
17/
FINAL
135
Game
Bird
Ration
Ingredients
Percent
Fine
Corn
Meal
44.83
Soy
Bean
Meal,
48%
Protein
30.65
Wheat
Midds
6.50
Protein
Base
6.00
Agway
Special,
60%
Protein
4.00
Alfalfa
Meal,
20%
Protein
3.00
Dried
Whey
2.50
Ground
Limestone
0.90
Eastman
CalPhos
0.60
Methionine
Premix
+
Liquid
0.35
Vitamin
and
Mineral
Premix
(
see
below)
0.32
GL
Form
(
Fermatco)
1
0.25
Salt
iodized
0.10
1Fermentation
by­
products
(
source
of
unidentified
growth
factors)

Vitamin
and
Mineral
Premix
Vitamin
or
Mineral
Amount
Per
Ton
Vitamin
D3
2,000,000
I.
C.
U.
Vitamin
A
7,000,000
I.
U.
Riboflavin
6
g
Niacin
40
g
Pantothenic
Acid
10
g
Vitamin
B12
8
mg
Folic
Acid
600
mg
Biotin
64
mg
Pyridoxine
1.2
g
Thiamine
1.2
g
Vitamin
E
20,000
I.
U.
Vitamin
K
(
Menadione
dimethylpyrimidinol
bisulfite)
5.8
g
Manganese
102
g
Zinc
47
g
Copper
6.8
g
Iodine
1.5
g
Iron
51
g
Selenium
182
g
Prophylaxis:
None
Brooding
compartment
mean
temperature:
38
±
2oC
Ambient
room
mean
temperature:
27.3
±
1.2oCAverage
relative
humidity:
31
±
14%
Photoperiod:
Sixteen
hours
light
per
day
Lighting:
fluorescent
lights
which
closely
approximate
noon­
day
sunlight;
average
of
approximately
139
lux
of
illumination
Test
diet
preparation:
Test
substance
mixed
directly
into
the
ration
by
means
of
a
Hobart
mixer.
No
ENV/
JM/
RD(
2002)
17/
FINAL
136
carrier
was
used.

Diet
sampling:
Homogeneity
of
the
test
substance
in
the
diet
evaluated
by
collecting
six
samples
from
the,
18.3
pprn
concentration
and
six
from
the
1171
ppm
concentration.
Samples
collected
from
the
top,
middle,
and
bottom
of
the
left
and
right
sections
of
the
mixing
vessel.
These
samples
also
served
as
the
verification
samples
for
these
concentrations.
Verification
samples
of
the
other
treatment
groups
(
two
samples
from
each)
and
the
control
(
one
sample)
were
collected
at
preparation
on
Day
0.
Stability
samples
were
collected
at
the
end
of
the
exposure
period
(
Day
5)
from
the
control
(
one
sample)
and
each
treatment
group
(
two
samples
each).

RESULTS
Nominal
concentrations:
Negative
control,
18.3,
36.6,
73.2,
146,
293,
586,
and
1171
ppm
Measured
concentrations:
<
LOQ,
19.5.
40.2,
74.5,
174,
291,
537,
and
1196
ppm
Element
value:
Dietary
LC50
=
220
(
164
­
289)
ppm
No
mortality
concentration
=
73.2
ppm
NOEC
(
body
weight
gain)
=
73.2
ppm
All
element
values
based
on
nominal
concentrations
Analytical
Methodology:
Diet
samples
were
extracted
with
methanol.
Analyses
of
test
solutions
were
performed
at
Wildlife
International
Ltd.
using
high
performance
liquid
chromatography
with
mass
spectrometric
detection
(
HPLC/
MS).
When
determining
the
concentration
of
the
test
substance
in
the
test
solutions,
the
same
and
most
prominent
peak
response
for
perfluorooctanesulfonate
was
used.
No
attempt
was
made
to
quantify
on
the
basis
of
individual
isomeric
components.
The
LOQ
(
limit
of
quantitation)
was
1.15
ppm
in
this
study.
The
mean
percent
recovery
of
matrix
fortifications
analyzed
concurrently
during
sample
analysis
was
94.7.
Samples
collected
for
determination
of
homogeneity
in
diet
ranged
from
102­
107%
of
nominal.
Samples
collected
for
verification
in
diet
had
measured
values
from
92
to
119%
of
nominal.
Measured
values
for
ambient
stability
samples
taken
at
Day
5
ranged
from
101
 
122%
of
nominal.

Summary
of
analytical
chemistry
data
Homogeneity
in
avian
diet
Nominal
Test
Concentration,
ppm
Measured
Values
at
Day
0,
ppm
Mean
Measured
Concentration,
ppm
Percent
of
Nominal
18.3
18.5,
23.4,
18.3,
17.3,
19.4,
19.9
19.5
107
1171
1239,
1221,
1118,
1301,
1163,
1133
1196
102
Verification
in
avian
diet
Nominal
Test
Concentration,
ppm
Measured
Duplicate
Concentrations
at
Day
0,
ppm
Mean
Measured
Concentration,
ppm
Percent
of
Nominal
Negative
Control
<
LOQ
­
­
36.6
46.7,
34.6
40.2
110
73.2
77.8,
71.2
74.5
102
146
176,
172
174
119
293
274,
307
291
99
586
550,
523
537
92
ENV/
JM/
RD(
2002)
17/
FINAL
137
Ambient
stability
in
avian
diet
Day
0
Mean
Measured
Concentration,
ppm
Measured
Duplicate
Concentrations
at
Day
5,
ppm
Mean
Measured
Concentration,
ppm
Mean
Percent
of
Day
0
Negative
Control
<
LOQ
­
19.5
19.2,
19.9
19.6
101
40.2
44.4,
53.8
49.1
122
74.5
76.4,
77.9
77.2
104
174
177,
174
176
101
291
318,
315
317
109
537
560,
665
613
114
1196
1260,
1187
1224
102
Biological
observations
Mortalities
and
clinical
observations:
One
incidental
mortality
occurred
in
the
control
group
as
a
result
of
a,
broken
leg
on
the
morning
of
Day
5.
It
was
subsequently
euthanized
on
Day
6.
Two
other
birds
in
the
control
group
were
intermittently
noted
with
foot
lesions
associated
with
cage
mate
aggression.
Otherwise,
all
control
birds
were
observed
to
be
normal
in
appearance
and
behavior
throughout
the
test.

The
first
treatment­
related
mortalities
occurred
on
Day
3
in
the
586
and
1171
ppm
treatment
groups.
Mortality
occurred
through
Day
8
in
all
dose
groups
>
146
ppm
with
some
of
the
deaths
being
during
the
post­
exposure
period.
There
were
no
treatment­
related
mortalities
or
overt
signs
of
toxicity
at
concentrations
<
73.2
ppm.

There
was
11
%
mortality
in
the
146
ppm
treatment
group,
and
two
additional
birds
displayed
clinical
signs
of
toxicity
(
wing
droop).
All
other
birds
in
this
test
group
displayed
normal
appearance
and
behavior
for
the
duration
of
the
test.
Recovery
with
normal
appearance
and
behavior
occurred
on
Day
9
to
test
termination.

There
was
80%
mortality
(
occurring
on
Days
5,
6,
and
7)
for
birds
in
the
293
ppm
treatment
group.
Signs
of
toxicity
observed
prior
to
death
included
a
ruffled
appearance,
reduced
reaction
to
stimuli
(
sound
and
motion),
lethargy,
wing
droop,
loss
of
coordination,
lower
limb
weakness
and
convulsions.
Recovery
with
normal
appearance
and
behavior
occurred
on
Day
9
to
test
termination.

There
was
100%
mortality
(
occurring
from
Day
3
through
Day
7)
for
birds
in
the
586
ppm
treatment
group.
Signs
of
toxicity
observed
prior
to
death
included
a
ruffled
appearance,
reduced
reaction
to
stimuli
(
sound
and
motion),
lethargy,
depression,
wing
droop,
loss
of
coordination,
lower
limb
weakness,
lower
limb
rigidity,
prostrate
posture,
and
convulsions.

There
was
100%
mortality
(
occurring
from
Day
2
(
noted
on
Day
3
for
Day
2
afternoon)
through
Day
4)
for
birds
In
the
1171
ppm
treatment
group.
Signs
of
toxicity
observed
prior
to
death
included
a
ruffled
appearance,
reduced
reaction
to
stimuli
(
sound
and
motion),
lethargy,
depression,
wing
droop,
loss
of
coordination,
lower
limb
weakness,
and
lower
limb
rigidity.

Body
weight
gain:
When
compared
to
the
control
group,
there
were
no
apparent
treatment
related
effects
on
body
weight
among
the
birds
in
concentrations
<
73.2
ppm.
During
Days
0­
5
statistically
significant
reductions
in
body
weight
gain
or
body
weight
loss
occurred
in
the
146,
293,
and
586
ppm
treatment
groups.
Body
weight
effects
could
not
be
determined
for
test
ENV/
JM/
RD(
2002)
17/
FINAL
138
organisms
in
the
1171
ppm
group
due
to
total
mortality.

Feed
Consumption:
No
apparent
treatment
related
effects
were
noted
for
feed
consumption
for
birds
in
concentrations
<
146
ppm.
Reduced
feed
consumption
was
noted
for
birds
In
treatment
groups
>
293
ppm
from
Days
0­
5.
No
treatment­
related
effects
on
feed
consumption
in
any
of
the
surviving
treatment
groups
during
the
Day
6­
8
post­
exposure
period
were
observed.

Gross
Necropsy:
All
birds
that
died
during
the
study,
half
of
those
surviving
at
Day
8
and
the
rest
at
test
termination
were
subjected
to
a
gross
necropsy.
Necropsy
results
for
birds
found
dead
were
similar,
including
thin
condition,
loss
of
muscle
mass,
altered
spleen
color,
autolysis
of
tissues
and
pale
organs.
These
necropsy
findings
were
considered
to
be
treatment
related.
The
single
bird
euthanized
from
the
293
ppm
treatment
was
found
to
have
treatment
related'
necropsy
findings.
Necropsy
results
for
all
other
birds
euthanized
on
Day
8
and
Day
22
were
unremarkable.

%
Cumulative
Mortality
Nominal
Concentration,
ppm
Day
1
Day
2
Day
3
Day
4
Day
5
Day
6
Day
7
Day
8*

Negative
Control
0
0
0
0
0
0
0
0
18.3
0
0
0
0
0
0
0
0
36.6
o
0
0
0
0
0
0
0
73.2
0
0
0
0
0
0
0
0
146
0
0
10**
0
0
0
10
10
293
0
0
0
0
20
40
80
80
586
0
0
10
20
50
80
100
100
1171
0
0
30
100
100
100
100
100
*
No
mortalities
occurred
in
any
of
the
treatment
levels
from
Day
8
to
Day
22
**
Bird
euthanized
on
day
3
after
sustaining
a
broken
log.

Body
Weight
(
grams)

Exposure
Period
Recovery
Period
Nominal
Concentration
(
ppm)
Mean
Body
Weight,
Day
0
Mean
Body
Weight
Change
Day
0­
5
Mean
Body
Weight
Change
Day
5­
8
Mean
Body
Weight
Change
Day
8­
15
Mean
Body
Weight
Change
Day
15­
22
Mean
Total
Body
Weight
Change,
Day
8­
22
Mean
Body
Weight,
Day
22
Negative
Control
20
+
10
+
8
+
23
+
22
45
82
18.3
21
+
11
+
9
+
24
+
23
47
87
36.6
20
+
11
+
8
+
26
+
24
50
89
73.2
20
+
9
+
7
+
24
+
20
44
79
146
20
+
7*
+
6**
+
24
+
21
45
79
293
20
­
2**
­
1**
+
14
+
20
34
55
586
20
­
4**
­
­
­
­­
­
1171
20
­
­
­
­
­

Note:
numbers
may
not
add
manually
due
to
rounding.
Values
for
293
ppm
treatment
group
are
impacted
by
the
fact
that
only
one
bird
remained
in
that
group
after
day
8.
ENV/
JM/
RD(
2002)
17/
FINAL
139
*
Statistically
different
from
the
control
group
at
p<
0.05
(
Dunnett's
t­
test)
**
Statistically
different
from
the
control
group
at
p=<
0.01
(
Dunnett's
t­
test)
(­)
=
no
data
available
due
to
mortality
Mean
average
feed
consumption
Nominal
Concentration,
ppm
Grams
Feed/
Bird/
Day
Days
0­
5
Grams
Feed/
Bird/
Day
Days
6­
8
Grams
Feed/
Bird/
Day
Days
8­
15
Grams
Feed/
Bird/
Day
Days
15­
22
Negative
Control
9
10
9
13
18.3
9
11
10
12
36.6
8
12
14
15
73.2
10
13
13
15
146
9
10
11
14
293
5
9
8
­
586
6
19
­
­
1171
4
­
­
­
(­)
=
No
data
available
due
to
mortality.

Gross
Pathological
Observations
from
Birds
that
Died
In
Study
Male,
Female,
and
Undetermined
(
ppm)
Finding
Control
N
=
l
146
N
=
2
293
N
=
8
586
N
=
10
1171
N
=
10
Abdominal
cavity,
some
autolysis
0
0
2
2
4
Abdominal
cavity,
autolysis
throughout
0
0
0
1
1
Crop,
empty
0
0
2
5
2
Emaciated
0
0
2
5
8
Fractured
leg
1
1
0
0
0
G.
I.
Tract,
empty
0
0
1
1
0
Gizzard
contents
bile
stained
0
0
2
5
1
Heart,
anterior
portion
mottled
white
color
0
0
1
0
0
Heart,
pale
.
0
0
0
2
1
Intestinal
contents
tar­
like
0
0
0
2
0
Keel,
prominent
0
0
1
3
10
Kidneys,
pale
0
0
0
2
0
Liver,
pale
and
mottled
0
1
0
0
0
Loss
of
muscle
mass
0
0
4
7
9
Muscular­
skeletal,
pale
0
1
0
0
0
Small
in
stature
0
0
3
0
0
Spleen,
black
0
0
0
1
0
Spleen,
dark
0
0
0
0
2
Spleen,
grey
0
0
0
1
0
Spleen,
grey­
brown
0
0
0
0
1
Spleen,
pale
0
0
1
0
1
Spleen,
small
0
0
0
0
1
Spleen,
small
and
pale
0
0
0
3
0
Thin
0
0
0
4
2
Not
remarkable
0
0
1
0
0
ENV/
JM/
RD(
2002)
17/
FINAL
140
CONCLUSIONS
The
dietary
LC50
value
for
Northern
Bobwhite
exposed
to
perfluorooctanesulfonate
was
determined
to
be
220
ppm
with
a
95%
confidence
Interval
of
164
to
289
ppm.
The
slope
of
the
concentration
response
curve
was
7.005
and
the
chi­
square
value
was
0.023.
The
no
mortality
concentration
was
73.2
ppm.
Based
upon
treatment
related
mortality,
signs
of
toxicity
and
effects
upon
body
weight
gain
at
the
146
ppm
test
concentration,
the
no
observed
effect
concentration
was
73.2
ppm.

Author
and/
or
submitter:
3M
Corporation,
Environmental
Laboratory,
P.
O.
Box
33331,
St.­
Paul,
Minnesota,
55133,
USA
DATA
QUALITY
Reliability:
Klimisch
ranking
=
1.

REFERENCES
This
study
was
conducted
at
Wildlife
International
Ltd.,
Easton,
MD
at
the
request
of
the
3M
company.

OTHER
Last
changed:
5/
3/
00
ENV/
JM/
RD(
2002)
17/
FINAL
141
Robust
Report
Reference
No.
13
­
Multi­
Phase
Exposure
/
Recovery
Algal
Assay
Test
TEST
SUBSTANCE
Identity:
Potassium
perfluorooetanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfbnic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks
field:
The
test
substance
is
a
white
powder
(
3M
Lot
583)
of
uncharacterized
purity.
The
following
summary
is
abbreviated
due
to
the
fact
that
this
study
has
been
superceded
by
a
more
recent
test.

METHOD
Method:
Test
protocol
utilized
was
modified
after
those
described
by
USEPA
­
600/
9­
78­
018,
1978;
ASTM­
E­
35.23,
1981;
OECD
201,1979.
ASTM
STP
#
667
Type:
Semi­
chronic
GLP:
No
Year
completed:
1981
Species:
Selenastrum
capricornutum
RESULTS
EC50
Values,
mg/
L
(
95%
confidence
interval
is
given
in
brackets)

Exposure
(
contact)
Days
Cell­
Dry
Weight
Cell­
Count
4
115
(
18
­
65)
82
(
No
Conf.
Limits)
7
122
(
41­
366)
99
(
19
­
398)
10
128
(
46
­
372)
98
(
16
­
431)
14
146
(
33
­
350)
95
(
12
­
455)

Remarks:
The
statistical
program
used
was
questionable.

DATA
QUALITY
Reliability:
Klimisch
ranking
=
2.
This
study
satisfied
criteria
for
quality
testing
at
the
time
performed,
but
actual
concentrations
were
not
measured.
Results
were
based
on
nominal
concentrations.
Additionally,
sample
purity
was
not
properly
characterized.

REFERENCES
This
study
was
conducted
by
the
3M
Company,
Environmental
Laboratory,
1981.

OTHER
Submitter:
3M
Company,
Environmental
Laboratory,
P.
O.
Box
33331,
St.
Paul,
Minnesota,
55133,
USA
Last
changed:
5/
3/
00
ENV/
JM/
RD(
2002)
17/
FINAL
142
Robust
Study
Report
Reference
No.
14
­
The
Effects
of
Continuous
Aqueous
Exposure
to
14C­
78.02
on
Hatchability
of
Eggs
and
Growth
and
Survival
of
Fry
of
Fathead
Minnow
(







	








	


/
Summary
of
histopathological
examinations
of
Fathead
Minnow
(







	








	


)
exposed
to
78.02
for
30
Days
TEST
SUBSTANCE
Identity:
Potassium
perfluorooctanesulfonate;
may
also
be
referred
to
as
14C­
78.02,
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
The
test
substance
is
a
white
powder.
Sample
was
radiolabeled.
Sample
purity
was
not
characterized.

The
following
summary
is
abbreviated
due
to
the
fact
that
this
study
has
been
superceded
by
a
more
recent
test.

METHOD
Method:
Method
was
developed
by
E
G
&
G,
Bionomic
and
closely
followed
those
presented
in
the
"
Proposed
recommended
bioassay
procedure
for
egg
and
fry
stages
of
freshwater
fish",
U.
S.
EPA,
1972.
Type:
Flow­
through
chronic
GLP:
No
Year
completed:
1978
Species:
Pimephales
promelas
RESULTS
30­
Day
NOEC:
1
mg/
L
30­
Day
LOEC:
1.9
mg/
L
30­
Day
MATC:
>
1
mg/
L
and
<
1.9
mg/
L
DATA
QUALITY
Reliability:
Klimisch
ranking
=
2.
This
study
satisfied
criteria
for
quality
testing
at
the
time
performed,
but
the
analytical
methodology
was
questionable.

REFERENCES
This
study
was
conducted
at
E
G
&
G,
Bionomics,
Aquatic
Toxicology
Laboratory
in
Wareham,
Massachusetts
at
the
request
of
the
3M
Company.

OTHER
Submitter:
3M
Company,
Environmental
Laboratory
P.
O.
Box
33331
St.
Paul,
MN
55133,
USA.

Last
changed:
5/
3/
00
ENV/
JM/
RD(
2002)
17/
FINAL
143
Robust
Study
Report
Reference
No.
15
­
Effect
of
Potassium
Perfluorooctanesulfonate
on
Survival,
etc.
(
Daphnid
reproduction)

TEST
SUBSTANCE
Identity:
Potassium
perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks
field:
The
test
substance
is
a
white
powder
of
uncharacterized
purity.

The
following
summary
is
abbreviated
due
to
the
fact
that
this
study
has
been
superceded
by
a
more
recent
test.

METHOD
Methods:
ASTM,
1981,
Proposed
Standard
Practice
for
Conducting
Renewal
Life­
Cycle
Toxicity
Tests
with
Daphnia
magna;
OECD,
1981,
Daphnia
sp.,
14­
day
Reproduction
Test.
Type:
Acute
static
and
Chronic
renewal
GLP:
No
Year
completed:
1984
Species:
Daphnia
magna
RESULTS
(
95%
confidence
interval
is
given
in
brackets)

48­
hour
EC50
(
immobilization):
27
(
25­
28)
mg/
L
14­
day
EC50
(
reproduction)*:
14.7(
12­
18)
mg/
L
21­
day
EC50
(
reproduction)*:
12.4(
11­
14)
mg/
L
28­
day
EC50
(
reproduction)*:
11.4(
10­
13)
mg/
L
28­
day
NOEC**:
7
mg/
L
14,
2
1
and
28­
day
MATC:
11.2
mg/
L
*
Cumulative
(
Young/
Adult)
**
This
concentration
applies
to
no
observed
effects
on
Cumulative
results
of
Young/
Adult
and
Broods/
Adult
and
on
the
total
number
of
Young/
Adult.

DATA
QUALITY
Reliability:
Klimisch
ranking
=
2.
This
study
satisfied
all
criteria
for
quality
testing
at
the
time
performed,
but
actual
concentrations
were
not
measured.
Results
were
based
on
nominal
concentrations.
Additionally,
sample
purity
was
not
adequately
characterized.

REFERENCES
This
study
was
conducted
by
the
3M
Company,
Environmental
Laboratory,
1984.

OTHER
Submitter:
3M
Company,
Environmental
Laboratory,
P.
O.
Box
33331,
St.
Paul,
Minnesota,
55133,
USA
Last
changed:
5/
3/
00
ENV/
JM/
RD(
2002)
17/
FINAL
144
Robust
Study
Report
Reference
No.
16
­







	








	


96­
hour
Toxicity
Test
Data
Summary.
Sample
FC­
94­
X
(
Li
salt
of
PFOS)

TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate,
Lithium
salt;
may
also
be
referred
to
as
PFOS
Li
salt,
FC­
94,
or
FC­
94­
X.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
lithium
salt,
CAS
#
29457­
72­
5)

Remarks:
Test
sample
was
taken
from
3M
production
lot
#
1.
The
test
sample
is
a
mixture
of
the
test
substance
in
water
(
approximately
24.5%
test
substance
and
75.5%
water).
No
calculations
were
made
to
adjust
for
the
actual
concentration
of
the
test
substance
in
the
test
sample.

METHOD
Method:
Not
noted.
Type:
Static
acute
GLP:
No
Year
completed:
1994
Species:
Pimephales
promelas
Supplier:
Aquatic
Biosystems
Inc.,
Fort
Collins,
CO,
USA
Analytical
monitoring:
pH
and
DO
content
Exposure
period:
96­
hours
Statistical
methods:
LC50
values
calculated
by
Trimmed
Spearman
­
Karber.
Test
fish
age:
79
days.
Length
and
weight:
Average
length
=
2.1
+
0.3cm
Average
weight
=
0.069
+
0.03
g
Loading:
0.69
g
fish
/
L
Pretreatment:
None
Test
Conditions
Dilution
water:
Carbon
filtered
well
water
Dilution
water
chemistry:
pH:
8.4
DO:
8.1
mg/
L
Stock
and
test
solution
preparation:
A
primary
stock
solution
was
prepared
in
dilution
water
to
yield
a
test
sample
concentration
of
400
mg/
L.
All
test
solutions
were
made
by
diluting
the
appropriate
amount
of
stock
solution
with
dilution
water
to
make
1
L
of
solution
per
concentration.
Stability
of
the
test
chemical
solutions:
Not
noted.
Exposure
vessels:
2
L
glass
beakers
Number
of
replicates:
two.
Number
of
fish
per
replicate:
ten
Number
of
concentrations:
six
plus
a
negative
control
Water
chemistry
during
the
study:
Dissolved
oxygen
range
(
0
 
96
hours):
6.0­
7.2
mg/
L
(
control
exposure)
4.8­
7.9
mg/
L
(
56.0
mg/
L
exposure)
pH
range
(
0
 
96
hours):
8.0­
8.4
(
control
exposure)
8.0­
8.4
(
56.0
mg/
L
exposure)
Test
temperature
range
(
0
 
96
hours):
19.2­
19.5oC
ENV/
JM/
RD(
2002)
17/
FINAL
145
RESULTS
Nominal
concentrations:
Negative
control,
3.2,
5.6,
10.0,
18.0,
32.0,
56.0
mg/
L
Element
value
(
95%
confidence
interval
is
given
in
brackets):
96­
hour
LC50
=
19
mg/
L
(
16­
24)
Mortality
of
controls:
None
Remarks:
Values
reported
are
for
the
test
sample.
No
calculations
were
made
to
adjust
for
the
concentration
of
the
test
substance
in
the
test
sample.

CONCLUSIONS
The
test
sample
containing
24.5%
Perfluorooctanesulfonate,
Lithium
salt
exhibited
a
96­
hour
LC50
for
fathead
minnow
of
19
mg/
L.
This
value
must
be
divided
by
4
in
order
to
express
the
result
in
terms
of
the
concentration
of
PFOS.

Submitter:
3M
Company,
Environmental
Laboratory
P.
O.
Box
33331
St.
Paul,
MN
55133,
USA.

DATA
QUALITY
Reliability:
Klimisch
ranking
2.
This
study,
while
well
conducted,
lacks
analytical
data
for:
determination
of
the
test
substance
concentration
in
the
test
solutions;
and
determination
of
the
sample
purity.

REFERENCES
This
study
was
conducted
by
the
3M
Company,
Environmental
Laboratory,
Lab
Request
number
M1018,
3/
25/
94.

OTHER
Last
changed:
7/
9/
01
ENV/
JM/
RD(
2002)
17/
FINAL
146
Robust
Study
Report
Reference
No.
17
­
48­
hour
Acute
Toxicity
to
Daphnia,













.
FC­
94­
X
(
Li
salt
of
PFOS)

TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate,
Lithium
salt;
may
also
be
referred
to
as
PFOS
Li
salt,
FC­
94,
or
FC­
94­
X.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
lithium
salt,
CAS
#
29457­
72­
5)

Remarks:
The
test
sample
is
a
mixture
of
the
test
substance
in
water
(
approximately
24.5%
test
substance
and
75.5%
water).
No
calculations
were
made
to
adjust
for
the
actual
concentration
of
the
test
substance
in
the
test
sample.

METHOD
Method:
Not
noted.
Test
type:
Static
acute
GLP:
No
Year
completed:
1994
Species:
Daphnia
magna
Analytical
monitoring:
pH
and
DO
content
Statistical
methods:
EC50
values
calculated
using
Trimmed
Spearman­
Karber
method
Test
daphnid
source:
Obtained
from
U.
S.
EPA­
NETAC,
Duluth,
Minnesota,
USA.
Test
daphnid
age
at
study
initiation:
<
24­
hours
Test
Conditions
Dilution
water:
Carbon­
filtered
well
water
Dilution
water
chemistry:
pH:
8.4
DO:
8.6
mg/
L
Stock
and
test
solutions
preparation:
A
primary
stock
solution
was
prepared
in
dilution
water
to
yield
a
test
sample
concentration
of
1000
mg/
L.
All
test
solutions
were
made
by
diluting
the
appropriate
amount
of
stock
solution
with
dilution
water
to
make
50
mL
of
solution
per
concentration.
Exposure
vessels:
100
mL
glass
beakers
containing
50
mL
of
test
solution.
Number
of
replicates:
4
Number
of
daphnids
per
replicate:
5
Number
of
concentrations:
five
plus
a
negative
control
Water
chemistry
during
the
study:
Dissolved
oxygen
at
test
termination:
7.0
mg/
L
(
control
exposure)
7.8
mg/
L
(
1000
mg/
L
exposure)
pH
at
test
termination:
8.6
(
control
exposure)
8.6
(
1000
mg/
L
exposure)
Test
temperature
range
(
0
 
48
hours)
20.1­
21.0
C
Element
basis:
mortality
and
immobilization
RESULTS
Nominal
concentrations:
Negative
control,
100,
180,
320,
560,
1000
mg/
L
ENV/
JM/
RD(
2002)
17/
FINAL
147
Element
value:
(
95%
confidence
interval
is
given
in
brackets)
24­
hour
EC50
=
330
(
290­
370)
mg/
L
48­
hour
EC50
=
210
(
190­
230)
mg/
L
48­
hour
NOEC
=
100
mg/
L
Statistical
Evaluation:
The
EC50
values
and
95%
confidence
intervals
were
calculated
using
the
Trimmed
Spearman­
Karber
method
with
trim
set
to
0%.
Mortality
of
controls:
None
Remarks:
Values
reported
are
for
the
test
sample.
No
calculations
were
made
to
adjust
for
the
concentration
of
the
test
substance
in
the
test
sample.

CONCLUSIONS
The
test
sample
containing
24.5%
Perfluorooctanesulfonate,
Lithium
salt
exhibited
a
48­
hour
EC50
for
Daphnia
magna
of
210
mg/
L.
This
value
must
be
divided
by
4
in
order
to
express
the
result
in
terms
of
the
concentration
of
PFOS.

Submitter:
3M
Company,
Environmental
Laboratory
P.
O.
Box
33331
St.
Paul,
MN
55133,
USA.

DATA
QUALITY
Reliability:
Klimisch
ranking
2.
This
study,
while
well
conducted,
lacks
analytical
data
for:
determination
of
the
test
substance
concentration
in
the
test
solutions;
and
determination
of
the
sample
purity.

REFERENCES
This
study
was
conducted
by
the
3M
Company,
Environmental
Laboratory,
Lab
Request
number
M1018,
2/
10/
94.

OTHER
Last
changed:
7/
9/
01
ENV/
JM/
RD(
2002)
17/
FINAL
148
Robust
Study
Report
Reference
No.
20
­
96­
hour
Acute
Toxicity
Test
on
Bluegill
Sunfish
(
FC­
99,
DEA
salt
of
PFOS)

TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate,
DEA
salt;
may
also
be
referred
to
as
PFOS
DEA
salt,
FC­
99,
or
3M
Sample
No.
2.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
compd.
with
2,2'­
iminobis[
ethanol]
(
1:
1),
CAS
#
70225­
14­
8)

Remarks:
Test
sample
is
a
mixture
of
the
test
substance
in
water
(
approximately
25%
test
substance
and
75%
water).
All
values
reported
relate
to
this
mixture.
No
calculations
were
made
to
adjust
for
the
actual
concentration
of
the
test
substance
in
the
test
sample.

METHOD
Method:
Environmental
Protection
Agency,
Ecological
Research
Series
EPA­
660/
3­
75­
009,
April,
1975.
Standard
Methods.
Type:
Static
acute
GLP:
Yes
Year
completed:
1979
Species:
Lepomis
macrochirus
Supplier:
Osage
Catfisheries,
Inc.
in
Osage
Beach,
Missouri,
USA.
Analytical
monitoring:
pH
and
DO
/
ammonia
content
Exposure
period:
96­
hours
Statistical
methods:
Probit
analysis.
Test
fish
age:
Not
noted.
Length
and
weight:
Average
length
=
28.6
+
2.17
mm.
Average
weight
=
0.60
+
0.15
g
Loading:
0.2
g
fish
/
L
Pretreatment:
None
Test
Conditions
Dilution
water:
Laboratory
well
water
Dilution
water
chemistry:
Dissolved
oxygen:
9.3
mg/
L
Hardness:
255
mg/
L
as
CaCO3
Alkalinity:
368
mg/
L
as
CaCO3
pH:
7.8
Conductivity:
50
µ
mhos/
cm
Stock
and
test
solution
preparation:
Primary
stock
prepared
in
deionized
water
at
a
concentration
of
150
mg/
mL.
The
test
concentrations
were
prepared
by
transferring
appropriate
aliquots
of
the
stock
standard
directly
to
the
test
chambers.
The
test
solutions
were
noted
to
foam
when
stirring
in
toxicant
aliquots.
Test
concentrations
were
prepared
based
on
total
sample,
not
on
percent
concentration
of
the
test
substance
in
the
test
sample.
Concentrations
dosing
rate:
Once
Stability
of
the
test
chemical
solutions:
Not
noted
Exposure
vessels:
40
liter
glass
aquaria
containing
30L
of
test
solution.
Number
of
replicates:
one
Number
of
fish
per
replicate:
ten
Number
of
concentrations:
six
plus
a
negative
control
ENV/
JM/
RD(
2002)
17/
FINAL
149
Water
chemistry
during
the
study:
Dissolved
oxygen
range
(
0
 
96
hours):
6.0
­
8.4
mg/
L
(
control
exposure)
5.8
­
8.3
mg/
L
(
18
mg/
L
exposure)
pH
range
(
0
 
96
hours):
8.2
­
8.3
(
control
exposure)
8.3
­
8.3
(
18
mg/
L
exposure)
Test
temperature:
Temperature
held
constant
at
22oC
through
use
of
a
water
bath
for
test
vessels.

RESULTS
Nominal
concentrations:
Negative
control,
18,
37,
75,
160,
320,
650
mg/
L
Element
value:
(
95%
confidence
interval
is
given
in
brackets)
24­
hour
LC50
=
460
(
370­
580)
mg/
L
48­
hour
LC50
=
370
(
290­
470)
mg/
L
96­
hour
LC50
=
31
(
22­
43)
mg/
L
96­
hour
NOEC
=
18
mg/
L
(
C.
I.
not
calculated)

All
element
values
based
on
nominal
concentrations
Statistical
evaluation
of
mortality:
Probit
analysis
was
used
to
calculate
LC50
values
and
the
corresponding
confidence
limits.

Quality
Check
for
Test
Organism
Health:
The
bluegill
sunfish
were
challenged
with
a
reference
compound,
Antimycin
A.
The
observed
96­
hour
LC50
and
95%
confidence
limits
(
C.
I.)
were
within
the
95%
confidence
limits
reported
in
the
literature,
indicating
that
the
fish
were
in
good
condition.

Cumulative
percent
mortality:

Nominal
Test
Concentration
mg/
L
24­
hours
48­
hours
72­
hours
96­
hours
Neg.
Control
0
0
0
0
18
0
0
0
0
37
0
0
10
80
75
0
0
30
90
160
0
0
70
100
320
0
20
100
100
650
100
100
100
100
CONCLUSIONS
The
test
sample
96­
hour
LC50
for
bluegill
sunfish
was
determined
to
be
31
mg/
L
with
a
95%
confidence
interval
of
22­
43
mg/
L.
The
96­
hour
no
observed
effect
concentration
was
18
mg/
L.
These
values
must
be
divided
by
4
in
order
to
express
the
results
in
terms
of
the
concentration
of
PFOS.
Submitter:
3M
Company,
Environmental
Laboratory
P.
O.
Box
33331
St.
Paul,
MN
55133,
USA.

DATA
QUALITY
Reliability:
Klimisch
ranking
2.
This
study,
while
well
conducted,
lacks
analytical
data
for
determination
of
the
test
substance
concentration
in
the
test
solutions
and
determination
of
the
sample
purity.
There
were
ENV/
JM/
RD(
2002)
17/
FINAL
150
also
gaps
in
the
measurement
of
water
quality
criteria
for
a
number
of
the
concentrations
at
given
time
intervals.

REFERENCES
This
study
was
conducted
by
Analytical
BioChemistry
Laboratories,
Inc.
of
Columbia,
Missouri
on
behalf
of
the
3M
Company.

OTHER
Last
changed:
5/
2/
00
ENV/
JM/
RD(
2002)
17/
FINAL
151
Robust
Study
Report
Reference
No.
23
 
Acute
toxicity
to
Aquatic
Invertebrates
(
e.
g.
Daphnia)

TEST
SUBSTANCE
Identity:
Potassium
perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95
or
as
part
of
the
mixed
product
FM­
3820
(
see
Remarks).
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
The
test
sample
is
FM­
3820,
a
mixture
of
the
test
substance
in
diethylene
glycol
butyl
ether
and
water
(
approximately
24­
28%
test
substance
in
diethylene
glycol
butyl
ether
and
water).
Calculations
were
made
to
adjust
test
values
using
the
upper
limit
concentration
of
the
test
substance
(
28%)
in
the
test
sample
and
no
adjustment
was
made
for
the
presence
of
the
diethylene
glycol
butyl
ether
or
water
when
noted
below.
These
calculations
assumed
that
all
toxicity
was
due
to
the
presence
of
the
Perfluorooctanesulfonate
substance.

METHOD
Method:
OECD
202
Test
type:
Static
acute
GLP:
Yes
Year
completed:
1991
Species:
Daphnia
magna
Analytical
monitoring:
DO,
pH,
Conductivity,
and
temperature
were
monitored
daily.
Statistical
methods:
EC50
values
calculated,
when
possible
by
standard
statistical
techniques
(
Stephan,
1983)
Test
daphnid
source:
Obtained
from
cultures
maintained
by
EnviroSystems
Division,
Resource
Analysts,
Inc.,
Hampton,
NH,
USA.
Test
daphnid
age
at
study
initiation:
<
24­
hours
Test
Conditions
Dilution
water:
Well
water
from
wells
at
EnviroSystems
in
Hampton,
New
Hampshire.
Dilution
water
chemistry:
pH:
7.8*
Conductivity:
1200
µ
mhos/
cm*
TOC:
<
2.0
mg/
L
*
Values
measured
at
time
of
test.

Lighting:
Cool
white
fluorescent
lights,
intensity
23
µ
E/
s/
m2.
Photoperiod
of
16­
hours
light,
8­
hours
dark.
No
transition
period
noted.
Stock
and
test
solutions
preparation:
A
primary
stock
solution
was
prepared
in
dilution
water
at
1000
mg/
L.
The
primary
stock
was
proportionally
diluted
with
dilution
water
to
prepare
the
five
test
concentrations.
Exposure
vessels:
250
mL
plastic
beakers
containing
200
mL
of
test
solution.
The
approximate
depth
of
test
solution
was
6
cm.
Number
of
replicates:
Four
Number
of
daphnids
per
replicate:
Five
Number
of
concentrations:
Five
plus
a
negative
control
Water
chemistry
during
the
study:
Dissolved
oxygen
range
(
0
 
48
hours):
8.2
 
8.5
mg/
L
(
control
exposure)
8.1
 
8.5
mg/
L
(
150
mg/
L
exposure)
ENV/
JM/
RD(
2002)
17/
FINAL
152
pH
range
(
0
 
48
hours):
7.8
 
8.6
(
control
exposure)
7.8
 
8.6
(
150
mg/
L
exposure)
Test
temperature
range
(
0
 
48
hours):
20.8
 
21.0oC
(
control
exposure)
20.7
 
20.9oC
(
150
mg/
L
exposure)

Conductivity
range
(
0
 
48
hours):
1200
 
1300
µ
mhos/
cm
(
control
exposure)
1200
 
1300
µ
mhos/
cm
(
150
mg/
L
exposure)

Element
basis:
mortality
RESULTS
Nominal
concentrations:
Negative
control,
25,
40,
60,
100,
150
mg/
L
Element
values:
24­
hour
EC50
=
>
150
mg/
L
(
C.
I.
not
calculable)
48­
hour
EC50
=
49
(
43­
56)
mg/
L
Perfluorooctanesulfonate
concentration
adjusted
element
value:
24­
hour
EC50
=
>
42
mg/
L
48­
hour
EC50
=
14
mg/
L
All
element
values
based
on
nominal
concentrations
Biological
observations:
Ninety
five
percent
survival
occurred
in
the
control
exposure.
The
number
of
surviving
organisms
and
the
occurrence
of
sub­
lethal
effects
and
immobilization
or
other
sublethal
effects
were
determined
visually
and
recorded
initially
and
after
24
and
48
hours.

Cumulative
percent
mortality:

Nominal
Test
Concentration
mg/
L
24­
hours
48­
hours
Neg.
Control
0
5
25
0
0
40
5
25
60
0
25
100
0
100
150
20
100
Control
response:
Satisfactory
CONCLUSIONS
The
test
substance
48­
hour
EC50
for
Daphnia
magna
was
determined
to
be
49
mg/
L
with
a
95%
confidence
interval
of
43­
56
mg/
L.
If
you
assume
all
toxicity
of
the
mixture
is
due
to
the
Perfluorooctanesulfonate,
the
adjusted
48­
hour
EC50
value
is
14
mg/
L
(
49
mg/
L
x
0.28).
Submitter:
3M
Company,
Environmental
Laboratory,
P.
O.
Box
33331
St.
Paul,
Minnesota
55133,
USA.

DATA
QUALITY
Reliability:
Klimisch
ranking
=
3.
The
study
lacks
analytical
measurement
of
test
substance
ENV/
JM/
RD(
2002)
17/
FINAL
153
concentrations
in
the
test
solutions
and
sample
purity
is
not
sufficiently
characterized.
Additionally,
data
are
for
a
mixture
and
toxicity
cannot
be
positively
attributed
to
PFOS
as
the
diethylene
glycol
butyl
ether
could
also
contribute
to
the
toxicity.
The
basic
water
quality
parameters
(
hardness,
alkalinity
and
calcium/
magnesium
ratio)
were
not
included
in
the
final
report.

REFERENCES
This
study
was
conducted
at
EnviroSystems
Division,
Resource
Analysts,
Incorporated,
Hampton,
NH,
USA
at
the
request
of
the
3M
Company.

OTHER
Last
changed:
5/
3/
00
ENV/
JM/
RD(
2002)
17/
FINAL
154
Robust
Study
Report
Reference
No.
26
 
Acute
toxicity
to
Fish
TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate,
Ammonium
salt;
may
also
be
referred
to
as
PFOS
NH4+
salt
or
FC­
93.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
ammonium
salt,
CAS
#
29081­
56­
9)

Remarks:
Test
sample
was
taken
from
3M
production
lot
#
1.
The
test
sample
is
a
mixture
of
the
test
substance
in
isopropanol
and
water
(
25%
test
substance,
20%
isopropanol,
55%
water).
No
calculations
were
made
to
adjust
for
the
actual
concentration
of
the
test
substance
in
the
test
sample.

METHOD
Method:
Not
noted.
Type:
Static
acute
GLP:
No
Year
completed:
1974
Species:
Pimephales
promelas
Supplier:
Not
noted.
Analytical
monitoring:
pH
and
DO
content
Exposure
period:
96­
hours
Statistical
methods:
Plotted
LC50
Test
fish
age:
Not
noted.
Length
and
weight:
Average
length
=
2
inches,
Average
weight
=
1.5
g
Loading:
Not
noted.
Pretreatment:
Not
noted
Test
Conditions
Dilution
water:
carbon
filtered
city
of
St.
Paul,
MN
water
Dilution
water
chemistry:
Not
noted.

Stock
and
test
solution
preparation:
Not
noted.
Concentrations
dosing
rate:
Once
Stability
of
the
test
chemical
solutions:
Not
noted.
Exposure
vessels:
Not
noted.
Number
of
replicates:
One.
Number
of
concentrations:
five
plus
a
negative
control
Water
chemistry
during
the
study:
Dissolved
oxygen
range
(
0
 
96
hours):
5.0­
5.9
mg/
L
(
control
exposure)
4.2­
5.0
mg/
L
(
100
mg/
L
exposure)
pH
range
(
0
 
96
hours):
7.0­
7.1
(
control
exposure)
7.0­
7.2
7.0­
7.1
(
100
mg/
L
exposure)
Test
temperature
range
(
0
 
96
hours):
21
 
22oC
(
70­
72oF)

RESULTS
Nominal
concentrations:
Negative
control,
10,
25,
50,
75,
100
mg/
L
ENV/
JM/
RD(
2002)
17/
FINAL
155
Element
value:
96­
hour
LC50
=
85
mg/
L
(
C.
I.
not
determined)
Mortality
of
controls:
None
Remarks:
95%
confidence
limits
were
not
calculated
for
this
material.
Additionally,
testing
was
conducted
on
the
mixture
of
the
test
substance
in
20%
isopropanol
and
55%
water.
The
value
reported
applies
to
that
mixture
and
not
the
test
substance.
No
attempt
was
made
to
determine
the
impact
of
the
presence
of
the
organic
solvent
or
what
portion
of
the
toxicity
can
be
contributed
to
the
Perfluorooctanesulfonate,
ammonium
salt.

CONCLUSIONS
The
test
sample
containing
25%
Perfluorooctanesulfonate,
ammonium
salt
exhibited
a
96­
hour
LC50
for
fathead
minnow
of
85
mg/
L.

Submitter:
3M
Company,
Environmental
Laboratory
P.
O.
Box
33331
St.
Paul,
MN
55133,
USA
DATA
QUALITY
Reliability:
Klimisch
ranking
3.

REFERENCES
This
study
was
conducted
by
the
3M
Company,
Environmental
Laboratory,
7/
29/
74
to
8/
2/
74.

OTHER
Last
changed:
5/
3/
00
ENV/
JM/
RD(
2002)
17/
FINAL
156
Robust
Study
Report
Reference
No.
27
 
Acute
toxicity
to
Fish
TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate,
Ammonium
salt;
may
also
be
referred
to
as
PFOS
NH4+
salt
or
FC­
93.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
ammonium
salt,
CAS
#
29081­
56­
9)

Remarks:
Test
sample
was
taken
from
3M
production
lot
#
1.
The
test
sample
is
a
mixture
of
the
test
substance
in
isopropanol
and
water
(
25%
test
substance,
20%
isopropanol,
55%
water).
No
calculations
were
made
to
adjust
for
the
actual
concentration
of
the
test
substance
in
the
test
sample.

METHOD
Method:
Not
noted.
Type:
Static
acute
GLP:
No
Year
completed:
1974
Species:
Pimephales
promelas
Supplier:
Not
noted.
Analytical
monitoring:
pH
and
DO
content
Exposure
period:
96­
hours
Statistical
methods:
Plotted
LC50.
Test
fish
age:
Not
noted.
Length
and
weight:
Average
length
=
2
inches,
Average
weight
=
1.5
g
Loading:
Not
noted.
Pretreatment:
Not
noted
Test
Conditions
Dilution
water:
Carbon
filtered
city
of
St.
Paul,
MN
water
Dilution
water
chemistry:
Not
noted.
Stock
and
test
solution
preparation:
Not
noted.
Concentrations
dosing
rate:
Once
Stability
of
the
test
chemical
solutions:
Not
noted.
Exposure
vessels:
Not
noted.
Number
of
replicates:
One.
Number
of
concentrations:
five
plus
a
negative
control
Water
chemistry
during
the
study:
Dissolved
oxygen
range
(
0
 
96
hours):
4.5­
5.7
mg/
L
(
control
exposure)
3.8­
5.0
mg/
L
(
125
mg/
L
exposure)
Not
recorded
at
highest
conc.
(
150
mg/
L)
due
to
100%
mortality.
pH
range
(
0
 
96
hours):
7.0­
7.0
(
control
exposure)
7.0­
7.0
(
125
mg/
L
exposure)
Not
recorded
at
highest
conc.
(
150
mg/
L)
due
to
100%
mortality.
Test
temperature
range
(
0
 
96
hours):
20
 
21oC
(
69­
70
oF)
ENV/
JM/
RD(
2002)
17/
FINAL
157
RESULTS
Nominal
concentrations:
Negative
control,
50,
75,
100,
125,
150
mg/
L
Element
value:
96­
hour
LC50
=
100
mg/
L
(
C.
I.
not
determined)
Mortality
of
controls:
None
Remarks:
95%
confidence
limits
were
not
calculated
for
this
material.
Additionally,
testing
was
conducted
on
the
mixture
of
the
test
substance
in
20%
isopropanol
and
55%
water.
The
value
reported
applies
to
that
mixture
and
not
the
test
substance.
No
attempt
was
made
to
determine
the
impact
of
the
presence
of
the
organic
solvent
or
what
portion
of
the
toxicity
can
be
contributed
to
the
Perfluorooctanesulfonate,
ammonium
salt.

CONCLUSIONS
The
test
sample
containing
25%
Perfluorooctanesulfonate,
ammonium
salt
exhibited
a
96­
hour
LC50
for
fathead
minnow
of
100
mg/
L
Submitter:
3M
Company,
Environmental
Laboratory
P.
O.
Box
33331
St.
Paul,
MN
55133,
USA
DATA
QUALITY
Reliability:
Klimisch
ranking
3.

REFERENCES
This
study
was
conducted
by
the
3M
Company,
Environmental
Laboratory,
10/
15/
74
to
10/
19/
74.

OTHER
Last
changed:
5/
3/
00
ENV/
JM/
RD(
2002)
17/
FINAL
158
Robust
Study
Report
Reference
No.
28
 
Acute
toxicity
of
P3025
Developmental
Material
to
Fathead
minnow
(







	








	


)

TEST
SUBSTANCE
Identity:
Perfluorooctylsulfonate,
didecyldimethylammonium
salt;
may
also
be
referred
to
as
Fluoroalkyl
ammonium
derivative.
[
1­
Decaminium,
N­
decyl­
N,
N­
dimethyl­,
salt
with
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­
1­
octanesulfonic
acid
(
1:
1),
CAS
#
251099­
16­
8]

Remarks:
The
3M
production
lot
number
was
Lot
1.
The
test
sample
is
L­
14394
referred
to
by
the
test
laboratory
as
P3025.
The
sample
was
labeled
F­
11615,
Lot
1.
The
test
sample
is
a
mixture
of
the
test
substance
in
water
(
approximately
30­
40%
test
substance,
60­
70%
water,
and
0­
5%
of
residual
perfluorochemicals).
All
values
reported
relate
to
this
mixture.
The
test
sample
appears
to
be
a
2­
phase
dispersion
(
clear
liquid
with
opaque
solid)
which
rapidly
separates
after
agitation.
No
calculations
were
made
to
adjust
for
the
actual
concentration
of
the
test
substance
in
the
test
sample.

METHOD
Method:
OECD
203
Type:
Static
acute
GLP:
No
Year
completed:
1996
Species:
Pimephales
promelas
Supplier:
Not
noted.
Analytical
monitoring:
DO,
pH,
temperature,
and
conductivity
were
monitored
daily.
Exposure
period:
96­
hours
Statistical
methods:
LL50
values
calculated
using
the
Trimmed
Spearman­
Karber
method.
The
NOEL
was
calculated
using
Fisher's
Exact
tests.
Test
fish
age:
Not
given.
Length
and
weight:
Average
length
=
11.3
mm,
Average
weight
=
7.8
mg
Loading:
0.26
g/
L
Pretreatment:
None
Test
Conditions
Dilution
water:
Dechlorinated
City
of
Duluth,
MN
tap
water.
Water
was
aerated
for
24­
hours
prior
to
use
in
the
test.
Dilution
water
chemistry:
Hardness:
48
mg/
L
as
CaCO3
pH:
8.08
Lighting:
Cool­
white
fluorescent
bulbs.
Photoperiod
of
16­
hours
light,
8­
hours
dark
used.
No
transition
period
noted.
Stock
and
test
solution
preparation:
Water
accommodated
fractions.
Test
solutions
were
prepared
individually
for
each
test
replicate
concentration
by
mass
addition
of
vigorously
shaken
test
substance
in
4
L
of
dilution
water.
The
solutions
were
vigorously
stirred
for
21­
hours
(
vortex
1/
2
to
1/
3
solution
depth).
The
aqueous
phase
was
siphoned
from
the
vessel
at
mid­
depth.

Concentrations
dosing
rate:
Once
Stability
of
the
test
chemical
solutions:
Not
noted.
Exposure
vessels:
4­
L
glass
jars
containing
3­
L
of
test
solution.
The
jars
were
sealed
with
Teflonlined
lids
fitted
with
stoppers
to
accommodate
oxygen
flushing
of
headspace.
Number
of
replicates:
two
ENV/
JM/
RD(
2002)
17/
FINAL
159
Number
of
fish
per
replicate:
ten
Number
of
concentrations:
three
plus
a
negative
control
Water
chemistry
during
the
study:
Dissolved
oxygen
range:
(
0
 
96
hours):
9.1
 
14.6
mg/
L
(
control
exposure)
8.7
 
18.2
mg/
L
(
700
mg/
L
exposure)
pH
range:
(
0
 
96
hours):
7.80
 
8.08
(
control
exposure)
7.78
 
7.99
(
700
mg/
L
exposure)
Test
temperature
range
(
0
 
96
hours):
20.8
 
20.9oC
Conductivity
range
(
0
 
96
hours):
128
 
142
µ
mhos/
cm
(
control
exposure)
118
 
154
µ
mhos/
cm
(
700
mg/
L
exposure)
Remarks:
Oxygen
was
added
to
the
headspace
in
the
jars
before
sealing
initially
and
at
each
observation
period.
The
dissolved
oxygen
concentrations
were
super­
saturated
in
the
test
vessels,
particularly
in
the
700
mg/
L
exposure
concentration.

RESULTS
Nominal
loading
concentrations:
Negative
control,
400,
700,
1,000
mg/
L.
Element
value:
(
95%
confidence
interval
is
given
in
brackets)
24­
hour
LL50
=
618
(
568
­
673)
mg/
L
48­
hour
LL50
=
607
(
554
­
664)
mg/
L
72­
hour
LL50
=
595
(
551
­
643)
mg/
L
96­
hour
LL50
=
562
(
523
­
604)
mg/
L
96­
hour
NOEL
=
<
490
mg/
L
All
element
values
based
on
nominal
concentrations.

Biological
observations
after
96­
hours:
No
mortality
or
abnormal
behavior
observed
in
the
negative
control
during
the
test.
Mortality
was
observed
in
the
remaining
exposure
concentrations.
Surfacing
was
observed
in
half
of
the
fish
at
the
700
mg/
L
exposure
concentration
at
24­
hours,
and
2
fish
were
quiescent
at
96­
hours.
No
abnormal
behavior
was
observed
in
the
400
mg/
L
exposure
concentration.

Cumulative
percent
mortality:

Nominal
Loading
Test
Concentration,
mg/
L
24­
hours
48­
hours
72­
hours
96­
hours
Neg.
Control
0
0
0
0
490
10
15
15
25
700
75
75
80
90
1,000
100
100
100
100
Lowest
concentration
causing
100%
mortality:
1,000
mg/
L
Mortality
of
controls:
None
Remarks:
Values
reported
are
for
the
test
sample.
No
calculations
were
made
to
adjust
for
the
concentration
of
the
test
substance
in
the
test
sample.

CONCLUSIONS
The
test
sample
96­
hour
LL50
for
fathead
minnow
was
determined
to
be
562
mg/
L
with
a
95%
confidence
interval
of
523
 
604
mg/
L.
The
96­
hour
no
observed
effects
level
(
NOEL)
was
<
490
mg/
L.
ENV/
JM/
RD(
2002)
17/
FINAL
160
Submitter:
3M
Company,
Environmental
Laboratory,
P.
O.
Box
33331,
St.
Paul,
Minnesota,
55133,
USA
DATA
QUALITY
Reliability:
Klimisch
ranking
2.
The
study
lacks
analytical
measurement
of
test
substance
concentrations
in
the
test
solutions
and
sample
purity
is
not
sufficiently
characterized.
Additionally,
data
is
for
a
mixture
and
toxicity
cannot
be
positively
attributed
to
didecyldimethylammonium
Perfluorooctylsulfonate
salt
alone.
Also,
supersaturation
of
the
test
solutions
with
oxygen
could
also
have
contributed
to
the
toxicity.

REFERENCES
This
study
was
conducted
at
AScI
Corporation,
Environmental
Testing
Division,
Duluth,
MN,
at
the
request
of
the
3M
Company.

OTHER
Last
changed:
5/
24/
00
ENV/
JM/
RD(
2002)
17/
FINAL
161
Robust
Study
Report
Reference
No.
29
 
Acute
toxicity
of
P3025
Developmental
Material












TEST
SUBSTANCE
Identity:
Perfluorooctylsulfonate,
didecyldimethylammonium
salt;
may
also
be
referred
to
as
Fluoroalkyl
ammonium
derivative.
[
1­
Decaminium,
N­
decyl­
N,
N­
dimethyl­,
salt
with
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­
1­
octanesulfonic
acid
(
1:
1),
CAS
#
251099­
16­
8]

Remarks:
The
3M
production
lot
number
was
Lot
1.
The
test
sample
is
L­
14394
referred
to
by
the
test
laboratory
as
P3025.
The
sample
was
labeled
F­
11615,
Lot
1.
The
test
sample
is
a
mixture
of
the
test
substance
in
water
(
approximately
30­
40%
test
substance,
60­
70%
water,
and
0­
5%
of
residual
perfluorochemicals).
All
values
reported
relate
to
this
mixture.
The
test
sample
appears
to
be
a
2­
phase
dispersion
(
clear
liquid
with
opaque
solid)
which
rapidly
separates
after
agitation.
No
calculations
were
made
to
adjust
for
the
actual
concentration
of
the
test
substance
in
the
test
sample.

METHOD
Method:
OECD
202
Test
type:
Static
acute
GLP:
No
Year
Completed:
1996
Species:
Daphnia
magna
Analytical
monitoring:
DO,
pH,
temperature
and
conductivity
were
monitored
daily.
Statistical
methods:
EL50
values
calculated
using
Trimmed
Spearman­
Karber
method.
NOEL
value
calculated
using
Steel's
Many­
One
Rank
test.
Test
daphnid
source:
Obtained
from
cultures
maintained
by
AScI
Corporation,
Duluth,
MN.
Test
daphnid
age
at
study
initiation:
<
24­
hours
Test
Conditions:
Dilution
water:
Dechlorinated
City
of
Duluth,
MN
tap
water.
Water
was
aerated
for
24­
hours
prior
to
use
in
the
test.
Dilution
water
chemistry:
Hardness:
44
mg/
L
as
CaCO3
pH:
8.04
Lighting:
Cool­
white
fluorescent
bulbs.
Photoperiod
of
16­
hours
light,
8­
hours
dark.
No
transition
period
noted.
Stock
and
test
solutions
preparation:
Water­
accommodated
fractions.
Test
solutions
were
prepared
individually
for
each
concentration
by
mass
addition
of
vigorously
shaken
test
substance
in
1
L
of
dilution
water.
The
solutions
were
vigorously
stirred
for
23­
hours
(
vortex
1/
2
to
1/
3
solution
depth).
The
aqueous
phase
was
siphoned
from
the
vessel
at
mid­
depth
after
settling
for
1­
hour.
Exposure
vessels:
250
mL
borosilicate
glass
beakers
containing
200
mL
of
test
solution.
The
solutions
were
kept
covered
during
the
test.
Number
of
replicates:
Four
Number
of
daphnids
per
replicate:
Five
Number
of
concentrations:
Five
plus
a
negative
control
Water
chemistry
during
the
study:
Dissolved
oxygen
range
(
0
 
48
hours):
8.6
 
9.1
mg/
L
(
control
exposure)
8.0
 
8.8
mg/
L
(
50
mg/
L
exposure)
ENV/
JM/
RD(
2002)
17/
FINAL
162
pH
range
(
0
 
48
hours):
8.04
 
8.11
(
control
exposure)
7.92
 
8.00
(
50
mg/
L
exposure)
Test
temperature
range
(
0
 
48
hours):
20.9
 
21.0oC
Conductivity
range
(
0
 
48
hours):
142
 
155
µ
mhos/
cm
(
control
exposure)
120
 
124
µ
mhos/
cm
(
50
mg/
L
exposure)
Element
basis:
mortality
and
immobilization
RESULTS
Nominal
loading
concentrations:
Negative
control,
3.13,
6.25,
12.5,
25,
50
mg/
L
Element
value:
(
95%
confidence
interval
is
given
in
brackets)
24­
hour
EL50
=
27.0
(
18.7­
39.0)
mg/
L
48­
hour
EL50
=
11.3
(
9.6­
13.2)
mg/
L
48­
hour
NOEL
=
6.25
mg/
L
All
element
values
based
on
nominal
concentrations.

Statistical
Evaluation:
The
EL50
values
and
95%
confidence
intervals
were
calculated
by
the
Trimmed
Spearman­
Karber
method.
The
NOEL
was
calculated
using
Steel's
Many­
One
Rank
test
using
the
TOXSTAT
statistical
software
Version
3.2,
University
of
Wyoming.

Biological
observations:
Daphnids
in
the
negative
control,
and
the
3.13
and
6.25
mg/
L
treatments
appeared
healthy
and
normal
throughout
the
test
with
no
mortality,
immobility
or
overt
clinical
signs
of
toxicity.
The
effects
noted
in
this
study
were
mortality;
no
immobilization
was
noted
at
any
test
concentration.
The
number
of
surviving
organisms
were
determined
visually
and
recorded
initially
and
after
24
and
48
hours.

Cumulative
percent
mortality:

Nominal
Loading
Test
Concentration
mg/
L
24­
hours
48­
hours
Negative
Control
0
0
3.13
0
0
6.25
0
0
12.5
5
70
25
50
95
50
75
100
Control
response:
satisfactory
Remarks:
Values
reported
are
for
the
test
sample.
No
calculations
were
made
to
adjust
for
the
concentration
of
the
test
substance
in
the
test
sample.

CONCLUSIONS
The
test
substance
48­
hour
EL50
for
Daphnia
magna
was
determined
to
be
11.3
mg/
L
with
a
95%
confidence
interval
of
9.6­
13.2
mg/
L.
The
48­
hour
no
observed
effect
level
(
NOEL)
was
6.25
mg/
L.

Submitter:
3M
Company,
Environmental
Laboratory
P.
O.
Box
33331
St.
Paul,
MN,
55133,
USA.
ENV/
JM/
RD(
2002)
17/
FINAL
163
DATA
QUALITY
Reliability:
Klimisch
ranking
2.
The
study
lacks
analytical
measurement
of
test
substance
concentrations
in
the
test
solutions
and
sample
purity
is
not
sufficiently
characterized.
Additionally,
data
is
for
a
mixture
and
toxicity
cannot
be
positively
attributed
to
didecyldimethylammonium
Perfluorooctylsulfonate
salt
alone.

REFERENCES
This
study
was
conducted
at
ASci
Corporation,
Environmental
Testing
Division,
Duluth,
MN,
at
the
request
of
the
3M
Company.

OTHER
Last
changed:
5/
24/
00
ENV/
JM/
RD(
2002)
17/
FINAL
164
Robust
Study
Report
Reference
No.
30
­
Acute
toxicity
of
PFOS
to
Rainbow
trout
in
saltwater
TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
The
test
substance
is
a
white
powder
of
uncharacterized
purity.

METHOD
Method:
Standard
procedures
for
Testing
Acute
Lethality
of
Liquid
Effluents
(
Environment
Canada,
1980)
Type:
Acute
static,
renewal
after
48­
hours
GLP:
No
Year
completed:
1985
Species:
Rainbow
Trout,
Salmo
gairdneri
Fish
source:
Rainbow
Springs,
Thamesford
Fish
age
at
test
initiation:
not
noted
Fish
acclimation
to
salt
water:
fish
gradually
acclimated
to
increasing
salinity;
held
at
30
parts
per
thousand
salinity
8
days
prior
to
test
initiation.
Exposure
period:
96­
hours
Analytical
monitoring:
Dissolved
oxygen,
pH,
conductivity
Statistical
methods:
Not
noted.
Element
values
were
calculated
for
each
replicate
series,
but
not
combined
for
the
whole
study.
A
cumulative
mortality­
concentration
plot
was
used
to
estimate
the
LC50.

Test
conditions:
Dilution
water:
Mississauga
dechlorinated
tap
water
amended
with
calcium,
magnesium,
sodium,
and
chloride
to
obtain
30
parts
per
thousand
salinity
Dilution
water
chemistry
(
initial):
pH:
7.4
­
8.0
D.
O.:
8.6
 
9.0
mg/
L
Conductivity:
>
20,000
µ
mhos/
cm
Stock
solution
preparation:
1000
mg/
L
Exposure
vessels:
Not
noted;
solution
volume
35
L
Number
of
replicates:
2
tests
 
run
4
days
apart,
not
replicated
Number
of
organisms/
vessel:
6
Loading:
0.75
g/
L
Number
of
concentrations:
4
plus
a
blank
control
Water
chemistry
during
the
studies:
Dissolved
oxygen
ranges
8.1
 
10.3
(
control)
8.8
 
10.1
(
30
mg/
L)
pH
ranges
7.6
 
8.2
(
control)
7.3
 
8.0
(
30
mg/
L)
Test
temperature
(
0
 
48
hours):
15
°
C
Photoperiod:
12­
hours
light,
12­
hours
dark
Element
basis:
mortality
ENV/
JM/
RD(
2002)
17/
FINAL
165
RESULTS
Nominal
concentrations:
5,
10,
20,
30
mg/
L
Element
values
(
95%
confidence
interval)
calculated
per
replicate:
96­
hour
LC50
=
13.7
(
10.7
 
17.7)
mg/
L
96­
hour
LC50
=
13.7
(
10.7
 
17.8)
mg/
L
Mortality
of
controls:
17%
(
1/
6
in
both
studies)

DATA
QUALITY
Reliability:
Klimisch
ranking
=
2.
This
study
satisfied
all
criteria
for
quality
testing
at
the
time
performed,
but
actual
concentrations
were
not
measured.
Results
were
based
on
nominal
concentrations.
Additionally,
sample
purity
was
not
adequately
characterized.

REFERENCES
This
study
was
conducted
by
Beak
Consultants
Limited,
Mississauga,
Ontario,
Canada
for
Panarctic
Oils
Ltd,
Calgary,
Alberta,
Canada.

OTHER
Submitter:
3M
Company,
Environmental
Laboratory,
P.
O.
Box
33331,
St.
Paul,
Minnesota,
55133
Last
changed:
7/
19/
01
ENV/
JM/
RD(
2002)
17/
FINAL
166
Robust
Study
Report
Reference
No.
31
­
Acute
toxicity
of
PFOS
to
Rainbow
trout
in
freshwater
TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
The
test
substance
is
a
white
powder
of
uncharacterized
purity.

METHOD
Method:
Standard
procedures
for
Testing
Acute
Lethality
of
Liquid
Effluents
(
Environment
Canada,
1980)
Type:
Acute,
renewal
after
48­
hours
GLP:
No
Year
completed:
1985
Species:
Rainbow
Trout
Fish
source:
Rainbow
Springs,
Thamesford
Fish
age
at
test
initiation:
not
noted
Exposure
period:
96­
hours
Analytical
monitoring:
Dissolved
oxygen,
pH,
conductivity
Statistical
methods:
Not
noted.
Element
values
were
calculated
for
each
replicate
series,
but
not
combined
for
the
whole
study.
A
cumulative
mortality­
concentration
plot
was
used
to
estimate
the
LC50.

Test
conditions:
Dilution
water:
Mississauga
dechlorinated
tap
water
Dilution
water
chemistry
(
initial):
pH:
7.5
­
8.5
D.
O.:
9.0
 
10.4
mg/
L
Stock
solution
preparation:
1000
mg/
L;
noted
as
cloudy
Exposure
vessels:
Not
noted;
solution
volume
35
L
Number
of
replicates:
2
tests
 
run
one
week
apart,
not
replicated
Number
of
organisms/
vessel:
6
Loading:
0.72
g/
L
Number
of
concentrations:
5
plus
a
blank
control,
4
plus
a
blank
control
Water
chemistry
during
the
studies:
Dissolved
oxygen
ranges
8.2
 
10.4
(
control)
8.0
 
9.5
(
30
mg/
L)
pH
ranges
7.4
 
8.3
(
control)
7.5
 
8.7
(
30
mg/
L)
Conductivity
range:
270
 
380
µ
mhos/
cm
Test
temperature
(
0
 
48
hours):
15
°
C
Element
basis:
mortality
RESULTS
Element
values
(
95%
confidence
interval)
calculated
per
replicate:
96­
hour
LC50
=
7.8
(
6.2
 
9.8)
mg/
L
ENV/
JM/
RD(
2002)
17/
FINAL
167
96­
hour
LC50
=
9.9
(
7.5
 
13.4)
mg/
L
Mortality
of
controls:
None
DATA
QUALITY
Reliability:
Klimisch
ranking
=
2.
This
study
satisfied
all
criteria
for
quality
testing
at
the
time
performed,
but
actual
concentrations
were
not
measured.
Results
were
based
on
nominal
concentrations.
Additionally,
sample
purity
was
not
adequately
characterized.

REFERENCES
This
study
was
conducted
by
Beak
Consultants
Limited,
Mississauga,
Ontario,
Canada
for
Panarctic
Oils
Ltd,
Calgary,
Alberta,
Canada.

OTHER
Submitter:
3M
Company,
Environmental
Laboratory,
P.
O.
Box
33331,
St.
Paul,
Minnesota,
55133
Last
changed:
7/
19/
01
ENV/
JM/
RD(
2002)
17/
FINAL
168
Robust
Study
Report
Reference
No.
32
­
Acute
toxicity
of
PFOS
to







sp.

TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
The
test
substance
is
a
white
powder
of
uncharacterized
purity.

METHOD
Method:
Draft
International
Standards
Organization
(
Vanhaecke
and
Persoone,
1981)
Type:
Acute
static
GLP:
No
Year
completed:
1985
Species:
Artemia
sp.
Artemia
source:
Salt
lake
Brine
Shrimp
Inc.
Artemia
age
at
test
initiation:
naupuli
<
24
hours
old.
Exposure
period:
48­
hours
Analytical
monitoring:
Dissolved
oxygen,
pH,
conductivity
Statistical
methods:
Not
noted.
Element
values
were
calculated
for
each
replicate
series,
but
not
combined
for
the
whole
study.
A
cumulative
mortality­
concentration
plot
was
used
to
estimate
the
LC50.
Test
conditions:
Dilution
water:
30
parts
per
thousand
NaCl
solution
Dilution
water
chemistry
(
initial):
pH:
8.0
 
8.2
D.
O.:
>
6
mg/
L
Stock
solution
preparation:
1000
mg/
L;
noted
as
cloudy
Exposure
vessels:
Not
noted;
solution
volume
10
mL
Number
of
replicates:
3
Number
of
organisms/
replicate:
10
Number
of
concentrations:
6
plus
a
blank
control
Water
chemistry
during
the
study:
Dissolved
oxygen
ranges
(
test
and
control):
>
6.0
mg/
L
pH
(
test
and
control)
8.0
 
8.2
Test
temperature
range
(
0
 
48
hours):
21
­
21
°
C
Element
basis:
mortality
RESULTS
Nominal
concentrations:
1,
2,
3,
5,
10,
20
mg/
L
Element
values
(
95%
confidence
interval)
calculated
per
replicate:
48­
hour
EC50
=
9.4
(
7.4
 
12.1)
mg/
L
48­
hour
EC50
=
9.4
(
7.3
 
12.2)
mg/
L
48­
hour
EC50
=
8.9
(
6.7
 
11.9)
mg/
L
Mortality
of
controls:
None
ENV/
JM/
RD(
2002)
17/
FINAL
169
DATA
QUALITY
Reliability:
Klimisch
ranking
=
2.
This
study
satisfied
all
criteria
for
quality
testing
at
the
time
performed,
but
actual
concentrations
were
not
measured.
Results
were
based
on
nominal
concentrations.
Additionally,
sample
purity
was
not
adequately
characterized.

REFERENCES
This
study
was
conducted
by
Beak
Consultants
Limited,
Mississauga,
Ontario,
Canada
for
Panarctic
Oils
Ltd,
Calgary,
Alberta,
Canada.

OTHER
Submitter:
3M
Company,
Environmental
Laboratory,
P.
O.
Box
33331,
St.
Paul,
Minnesota,
55133
Last
changed:
6/
12/
01
ENV/
JM/
RD(
2002)
17/
FINAL
170
Robust
Study
Report
Reference
No.
33
­
Acute
toxicity
of
PFOS
to













TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
The
test
substance
is
a
white
powder
of
uncharacterized
purity.
The
following
summary
is
abbreviated
due
to
the
fact
that
this
study
has
been
superceded
by
a
more
recent
test.

METHOD
Method:
International
Standards
Organization
(
1982)
Type:
Acute
static
GLP:
No
Year
completed:
1985
Species:
Daphnia
magna
RESULTS
Nominal
concentrations:
10,
20,
30,
50,
100
mg/
L
Number
of
replicates:
2
Element
values
(
95%
confidence
interval)
calculated
per
replicate
48­
hour
EC50
=
58
(
46
 
72)
mg/
L
48­
hour
EC50
=
67
(
48
 
92)
mg/
L
DATA
QUALITY
Reliability:
Klimisch
ranking
=
2.
This
study
satisfied
all
criteria
for
quality
testing
at
the
time
performed,
but
actual
concentrations
were
not
measured.
Results
were
based
on
nominal
concentrations.
Additionally,
sample
purity
was
not
adequately
characterized.

REFERENCES
This
study
was
conducted
by
Beak
Consultants
Limited,
Mississauga,
Ontario,
Canada
for
Panarctic
Oils
Ltd,
Calgary,
Alberta,
Canada.

OTHER
Submitter:
3M
Company,
Environmental
Laboratory,
P.
O.
Box
33331,
St.
Paul,
Minnesota,
55133
Last
changed:
5/
26/
01
ENV/
JM/
RD(
2002)
17/
FINAL
171
Robust
Study
Report
Reference
No.
34
­
Perfluorooctanesulfonate,
Potassium
salt
(
PFOS):
An
acute
oral
toxicity
study
with
the
Honey
bee
TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks
field:
The
3M
production
lot
number
was
217.
The
test
substance
is
a
white
powder.
Sample
was
stored
AT
16­
20oC
prior
to
testing.
Purity
determined
to
be
86.9%
by
LC/
MS,
1H­
HMR,
19F­
NMR
and
elemental
analyses
techniques.

METHOD
Method:
OECD
Guideline
213,
EPPO
Guideline
170
Test
type:
Acute
Oral
GLP:
Yes
Year
Completed:
2001
Species:
Apis
mellifera
L.
Analytical
monitoring:
None
 
nominal
concentrations
Test
honey
bee
source:
Obtained
from
colony
number
32
belonging
to
the
Central
Science
Laboratory
(
CSL),
Sand
Hutton,
York,
UK,
National
Bee
Unit.
Test
honey
bee
age
at
study
initiation:
Young
adult
Test
honey
bee
type:
Worker
honey
bees,
free
of
acarine,
nosema
and
amoeba.
Varroacide
treatment:
None
within
the
4
weeks
prior
to
test
initiation.

Test
conditions
Humidity:
65%
+
5%
Temperature:
25
+
2oC
Lighting:
Conducted
in
darkness
Stock
and
test
solutions
preparation:
Test
substance:
Initial
stock
solution
prepared
in
analytical
grade
acetone
to
a
final
concentration
of
47.8
µ
g
PFOS/
µ
L
(
nominal
concentration).
Final
test
concentrations
prepared
from
dilutions
of
this
solution
with
50%
w/
v
sucrose.
Resulting
acetone
concentration
was
5%.
Reference
toxicant:
Primary
stock
solution
of
dimethoate
was
prepared
in
deionized
water
containing
1
g/
L
Triton
X­
100
to
a
final
concentration
of
3.0
µ
g/
µ
L.
Secondary
stock
solutions
were
made
by
diluting
the
primary
stock
solution
in
deionized
water
containing
1
g/
L
Triton
X­
100.
Final
test
concentrations
prepared
from
dilutions
of
these
solutions
with
50%
w/
v
sucrose.
Stability
of
the
test
chemical
solution:
A
dispersion
test
was
carried
out
on
an
86
µ
g
PFOS/
µ
L
acetone
solution
before
the
toxicity
study
was
performed.
The
homogeneity
of
the
mixture
was
assessed
after
2
hours.
The
test
item
formed
a
clear
solution
on
mixing;
after
2
hours
at
room
temperature,
slight
sediment
was
noted.
For
the
toxicity
test,
all
solutions
were
re­
mixed
prior
to
use.
The
contract
laboratory
considered
the
solutions
of
the
test
doses
to
be
homogenous
for
the
purpose
of
administration.
Exposure
vessels:
Clean,
well­
ventilated,
inverted
petri
dishes,
measuring
approximately
9
cm
in
diameter.
Feeding:
During
the
first
four
hours
of
the
test,
bees
provided
with
50%
w/
v
aqueous
sucrose
solutions
containi
ng
the
appropriate
PFOS
dose.
After
4­
hours,
dosed
sucrose
removed,
and
bees
provided
with
50%
w/
v
aqueous
sucrose
solutions,
continuously
available
through
the
end
ENV/
JM/
RD(
2002)
17/
FINAL
172
of
the
exposure
period.
Number
of
replicates:
Three
Number
of
bees
per
replicate:
Ten
Negative
control:
50%
w/
v
sucrose
Solvent
control:
50%
w/
v
sucrose
plus
5%
acetone
Reference
substance:
Dimethoate
Reference
substance
control:
Triton
X­
100
Number
of
concentrations:
five
plus
a
negative
and
a
solvent
control
Dose
administration:
The
bees
were
anaesthetized
with
carbon
dioxide
immediately
before
dosing
and
gently
tipped
out
onto
filter
paper
and
counted
into
the
petri
dish
cage
(
drones
were
discarded).
Each
group
of
10
bees
was
offered
0.2
mL
of
a
given
test
concentration
or
control
solution.
The
dose
was
measured
into
a
small,
pre­
weighed,
glass
feeder
within
the
cage
using
a
variable
volume
pipette.
This
volume
of
solution
is
equivalent
to
20
µ
L
per
bee.
Dose
frequency:
Once,
for
4
hours
of
exposure
Dose
calculation:
Feeders
were
weighed
after
removal
from
the
cages
to
determine
the
dose
consumed
per
bee.
Element
basis:
Mortality
RESULTS
Nominal
concentrations:
Negative
control
(
sucrose
only),
acetone
+
sucrose
control,
0.205,
0.450,
0.991,
2.17,
4.78
µ
g/
bee
Element
value
and
95%
confidence
interval:
24­
hour
LD50
=
0.72
(
0.60
 
0.85)
µ
g/
bee
48­
hour
LD50
=
0.46
(
0.32
 
0.55)
µ
g/
bee
72­
hour
LD50
=
0.40
(
0.33
 
0.48)
µ
g/
bee
72­
hour
NOEL
=
0.21
µ
g/
bee
All
element
values
based
on
nominal
concentrations
Statistical
Evaluation:
Probit
mortality
plotted
against
the
logarithm
of
dose
using
the
contract
laboratory
Probit
1
package.
A
least­
squares
regression
(
Finney
1971)
was
fitted
to
these.
The
NOELs
were
estimated
using
Student's
t­
test
(
p<
0.05)

Biological
observations:
There
was
significant
mortality
at
all
doses
above
a
mean
intake
of
0.21
µ
g/
bee
with
a
steep
dose
response
between
mean
intakes
of
0.45
and
2.2
µ
g/
bee.

Cumulative
percent
mortality:
Nominal
Test
Conc.,
µ
g/
bee
4­
hours
24­
hours
48­
hours
72­
hours
Negative
Control
0
0
0
0
Solvent
Control
0
3.3
3.3
3.3
0.205
0
0
6.7
10
0.450
0
20
50
60
0.991
0
70
93
97
2.17
6.7
100
100
100
4.78
30
100
100
100
ENV/
JM/
RD(
2002)
17/
FINAL
173
Sub­
lethal
Effects
 
Percent
Knockdown
(
K)
or
Stumbling
(
S):
Nominal
Test
Conc.,
µ
g/
bee
4­
hours
24­
hours
48­
hours
72­
hours
Negative
Control
0
0
0
0
Solvent
Control
0
0
0
0
0.205
0
0
0
0
0.450
0
3.3
(
S)
3.3
(
K)
3.3
(
K)
0.991
0
0
0
0
2.17
0
0
0
0
4.78
10
(
K)
0
0
0
Control
response:
satisfactory
Reference
toxicant
response:
satisfactory
 
dimethoate
72­
hour
LD50
=
0.11
µ
g/
bee
CONCLUSIONS
The
potassium
perfluorooctanesulfonate
72­
hour
oral
LD50
for
the
honey
bee
was
determined
to
be
0.40
µ
g/
bee
with
a
95%
confidence
interval
of
0.33
 
0.48.
The
72­
hour
no
observed
effect
level
was
0.21
µ
g/
bee.
The
dose
response
was
steep
between
a
mean
uptake
of
0.45
and
2.2
µ
g/
bee.

Submitter:
3M
Company,
Environmental
Laboratory,
P.
O.
Box
33331,
St.
Paul,
Minnesota,
55133
DATA
QUALITY
Reliability:
Klimisch
ranking
1
REFERENCES
This
study
was
conducted
at
Central
Science
Laboratory,
Sand
Hutton,
York,
UK,
under
contract
by
Wildlife
International,
Ltd,
Easton,
MD
at
the
request
of
the
3M
Company,
Lab
Request
Number
U2723,
2001.

OTHER
Last
changed:
5/
1/
01
ENV/
JM/
RD(
2002)
17/
FINAL
174
Robust
Study
Report
Reference
No.
35
­
Perfluorooctanesulfonate,
Potassium
salt
(
PFOS):
An
acute
contact
toxicity
study
with
the
Honey
bee
TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks
field:
The
3M
production
lot
number
was
217.
The
test
substance
is
a
white
powder.
Sample
was
stored
AT
16­
20oC
prior
to
testing.
Purity
determined
to
be
86.9%
by
LC/
MS,
1H­
HMR,
19F­
NMR
and
elemental
analyses
techniques.

METHOD
Method:
USEPA
OPPTS
850.3020
(
draft),
OECD
Guideline
214,
EPPO
Guideline
170
Test
type:
Acute
Contact
GLP:
Yes
Year
Completed:
2001
Species:
Apis
mellifera
L.
Analytical
monitoring:
None
 
nominal
concentrations
Test
honey
bee
source:
Obtained
from
colony
number
32
belonging
to
the
Central
Science
Laboratory
(
CSL),
Sand
Hutton,
York,
UK,
National
Bee
Unit.
Test
honey
bee
age
at
study
initiation:
Young
adult
Test
honey
bee
type:
Worker
honey
bees,
free
of
acarine,
nosema
and
amoeba.
Varroacide
treatment:
None
within
the
4
weeks
prior
to
test
initiation.

Test
conditions
Humidity:
65%
+
5%
Temperature:
25
+
2oC
Lighting:
Conducted
in
darkness
Stock
and
test
solutions
preparation:
Test
substance:
Stock
solution
prepared
in
analytical
grade
acetone
to
a
final
concentration
of
90.4
µ
g
PFOS/
µ
L
(
nominal
concentration).
Final
test
concentrations
prepared
in
acetone
from
dilutions
of
this
solution.
Reference
toxicant:
Stock
solution
of
dimethoate
was
prepared
in
deionized
water
containing
1
g/
L
Triton
X­
100
to
a
final
concentration
of
3.0
µ
g/
µ
L.
Final
test
concentrations
prepared
from
dilutions
of
this
solution.
Stability
of
the
test
chemical
solution:
A
dispersion
test
was
carried
out
on
an
86
µ
g
PFOS/
µ
L
acetone
solution
before
the
toxicity
study
was
performed.
The
homogeneity
of
the
mixture
was
assessed
after
2
hours.
The
test
item
formed
a
clear
solution
on
mixing;
after
2
hours
at
room
temperature,
slight
sediment
was
noted.
For
the
toxicity
test,
all
solutions
were
re­
mixed
prior
to
use.
The
contract
laboratory
considered
the
solutions
of
the
test
doses
to
be
homogenous
for
the
purpose
of
administration.
Exposure
vessels:
Clean,
well­
ventilated,
inverted
petri
dishes,
measuring
approximately
9
cm
in
diameter.
Feeding:
50%
w/
v
aqueous
sucrose
solution,
continuously
available
Number
of
replicates:
Three
Number
of
bees
per
replicate:
Ten
Negative
control:
Undosed
Solvent
control:
Acetone
Reference
substance:
Dimethoate
ENV/
JM/
RD(
2002)
17/
FINAL
175
Reference
substance
control:
Triton
X­
100
Number
of
concentrations:
five
plus
a
negative
and
a
solvent
control
Dose
administration:
The
bees
were
anaesthetized
with
carbon
dioxide
immediately
before
dosing
and
gently
tipped
out
onto
filter
paper
and
counted
into
the
petri
dish
cage
(
drones
were
discarded).
Each
bee
was
dosed
on
the
thorax
with
a
1
µ
L
drop
of
a
given
test
item
concentration
or
1
µ
L
acetone
before
being
placed
into
the
test
chamber.
Dose
frequency:
Once
Element
basis:
Mortality
RESULTS
Nominal
concentrations:
Negative
control,
acetone
control
(
1.0
µ
/
bee),
1.93,
4.24,
9.30,
20.5,
45,
µ
g
PFOS/
bee
Element
value
and
95%
confidence
interval:
24­
hour
LD50
=
38.9
(
28.2
 
71.2)
µ
g/
bee
48­
hour
LD50
=
10.4
(
8.2
 
13.0)
µ
g/
bee
72­
hour
LD50
=
6.0
(
4.7
 
7.6)
µ
g/
bee
96­
hour
LD50
=
4.78
(
3.8
 
5.8)
µ
g/
bee
96­
hour
NOEL
=
1.93
µ
g/
bee
All
element
values
based
on
nominal
concentrations
Statistical
Evaluation:
Probit
mortality
plotted
against
the
logarithm
of
dose
using
the
contract
laboratory
Probit
1
package.
A
least­
squares
regression
(
Finney
1971)
was
fitted
to
these.
The
NOELs
were
estimated
using
Student's
t­
test
(
p<
0.05)

Biological
observations:
There
was
significant
mortality
at
all
doses
above
1.93
µ
g/
bee
with
a
steep
dose
response
between
4.24
and
9.30
µ
g/
bee
Cumulative
percent
mortality:
Nominal
Test
Conc.,
µ
g/
bee
4­
hours
24­
hours
48­
hours
72­
hours
96­
hours
Negative
Control
0
0
3.3
3.3
3.3
Solvent
Control
3.3
3.3
3.3
3.3
3.3
1.93
0
6.7
6.7
13
13
4.24
0
0
13
37
37
9.30
0
6.7
40
63
90
20.5
0
40
93
97
100
45.0
0
50
93
100
100
Sub­
lethal
Effects
 
Percent
Knockdown
(
K)
or
Stumbling
(
S):
Nominal
Test
Conc.,
µ
g/
bee
4­
hours
24­
hours
48­
hours
72­
hours
96­
hours
Negative
Control
0
0
0
0
0
Solvent
Control
0
0
0
0
0
1.93
3.3
(
K)
0
0
0
0
4.24
0
0
0
0
0
9.30
0
0
0
0
3.3
(
K)
20.5
0
3.3
(
K)
0
3.3
(
K)
0
45.0
0
3.3
(
K)
3.3
(
K)
0
0
ENV/
JM/
RD(
2002)
17/
FINAL
176
Control
response:
satisfactory
Reference
toxicant
response:
satisfactory
 
dimethoate
96­
hour
LD50
=
0.19
µ
g/
bee
CONCLUSIONS
The
potassium
perfluorooctanesulfonate
96­
hour
contact
LD50
for
the
honey
bee
was
determined
to
be
4.78
µ
g/
bee
with
a
95%
confidence
interval
of
3.8
 
5.8.
The
96­
hour
no
observed
effect
level
was
1.93
µ
g/
bee.
The
dose
response
was
steep
between
4.24
and
9.30
µ
g/
bee.

Submitter:
3M
Company,
Environmental
Laboratory,
P.
O.
Box
33331,
St.
Paul,
Minnesota,
55133
DATA
QUALITY
Reliability:
Klimisch
ranking
1
REFERENCES
This
study
was
cond
ucted
at
Central
Science
Laboratory,
Sand
Hutton,
York,
UK,
under
contract
by
Wildlife
International,
Ltd,
Easton,
MD
at
the
request
of
the
3M
Company,
Lab
Request
Number
U2723,
2001.

OTHER
Last
changed:
5/
1/
01
ENV/
JM/
RD(
2002)
17/
FINAL
177
Robust
Study
Report
Reference
No.
36
­
PFOS:
A
96­
hour
toxicity
test
with
the
freshwater
alga
(










	








)

TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
Sample
from
3M
production
lot
number
217.
The
test
substance
is
a
white
powder.
Purity
determined
to
be
86.9%
by
LC/
MS,
1H­
HMR,
19F­
NMR
and
elemental
analyses
techniques.

METHOD
Method:
OPPTS
850.5400
Test:
Acute
static
GLP:
Yes
Year
completed:
2001
Species:
Anabaena
flos­
aquae
Source:
Originally
from
UTEX
 
The
Culture
Collection
of
Algae
at
the
university
of
Texas
at
Austin,
and
maintained
in
culture
medium
at
Wildlife
International
Ltd.,
Easton,
MD
Analytical
monitoring:
PFOS
measured
at
0,
72,
96­
hours
Element
basis:
Reported
three
ways:
number
of
cells/
ml,
area
under
the
growth
curve
and
growth
rate
Exposure
period:
96­
hours
Start
date:
1/
28/
00
End
date:
6/
5/
00
Test
organisms
laboratory
culture:
Algae
cultures
had
been
actively
growing
in
algal
culture
medium
for
at
least
two
weeks
prior
to
test
initiation.
Stock
nutrient
solutions
were
prepared
by
adding
reagent­
grade
chemicals
to
reverse
osmosis­
purified
well
water.
Solutions
were
then
diluted
in
purified
well
water
to
prepare
final
growth
media.
Test
Conditions:
Freshwater
Algal
medium
Compound
Nominal
Concentration
Units
MgCl2.6H2O
12.16
mg/
L
CaCl2.2H2O
4.40
mg/
L
H3BO3
0.1856
mg/
L
MnCl2.4H2O
0.416
mg/
L
ZnCl2
3.28
µ
g/
L
FeCl3.6H20
0.1598
mg/
L
CoCl2.6H20
1.428
µ
g/
L
Na2MoO4.2H20
7.26
µ
g/
L
CuCl2.2H2O
0.012
µ
g/
L
Na2EDTA.
2H20
0.300
mg/
L
NaNO3
25.5
mg/
L
MgSO4.7H2O
14.7
mg/
L
K2HPO4
1.044
mg/
L
NaHCO3
15.0
mg/
L
Dilution
water
source:
The
pH
of
the
medium
was
adjusted
to
7.5
+
0.1
and
it
was
sterilized
by
ENV/
JM/
RD(
2002)
17/
FINAL
178
filtration
(
0.22
µ
m)
prior
to
use.
Test
solution
preparations:
Individual
test
solutions
were
prepared
in
algal
medium
at
each
of
the
six
nominal
concentrations.
The
solutions
were
stirred
with
magnetic
stir
plates
for
approximately
18
hours.
The
final
test
solutions
appeared
clear
and
colorless.
Exposure
vessels:
Sterile
250
mL
glass
Erlenmeyer
flasks
plugged
with
foam
stoppers
containing
100
mL
of
test
solution.
Agitation:
Shaken
continuously
at
100
rpm
Number
of
replicates:
six
(
including
3
for
analysis
of
exposure
concentration).
Initial
algal
cell
loading:
1.0
X
104
cells/
mL
Cell
counts
method:
hemacytometer
and
microscope
Number
of
concentrations:
six
plus
a
negative
control
plus
an
abiotic
control
at
the
highest
concentration
tested
Water
chemistry:
pH
range
(
0
 
96
hours)
7.4
 
7.6
(
control
exposure)
7.4
 
7.4
(
329
mg/
L
exposure)
Test
temperature
range
(
0
 
96
hours)
22.8
 
23.8
º
C
Light
levels:
(
0
 
96
hours)
1990
 
2310
lux
from
cool­
white
fluorescent
lighting
Photoperiod:
24­
hours
light
Method
of
calculating
mean
measured
concentrations:
arithmetic
mean
obtained
using
results
obtained
at
0­
hours,
72­
hours
and
96­
hours
RESULTS
Nominal
concentrations:
Negative
control,
37.9,
58.6,
88.8,
139,
216,
331
mg/
L
plus
331
abiotic
replicate
Measured
concentrations:
<
LOQ,
37.9,
63.9,
93.8,
143,
235,
329
mg/
L;
abiotic
replicate
=
349
mg/
L
Element
values
(
95%
confidence
interval):
24­
hour
EC50
(
cell
density)
=
105
mg/
L
(
C.
I.
not
calculable)
24­
hour
EbC50
(
area
under
curve)
=
90
(
40
­
150)
mg/
L
24­
hour
ErC50
(
growth
rate)
=
94
(
33
­
145)
mg/
L
48­
hour
EC50
(
cell
density)
=
117
mg/
L
(
C.
I.
not
calculable)
48­
hour
EbC50
(
area
under
curve)
=
103
mg/
L
(
C.
I.
not
calculable)
48­
hour
ErC50
(
growth
rate)
=
128
mg/
L
(
C.
I.
not
calculable)
72­
hour
EC10
(
cell
density)
=
43
(
34
 
84)
mg/
L
72­
hour
EbC10
(
area
under
curve)
=
<
38
mg/
L
(
C.
I.
not
calculable)
72­
hour
ErC10
(
growth
rate)
=
82
(
49
­
116)
mg/
L
72­
hour
EC50
(
cell
density)
=
120
(
92
­
139)
mg/
L
72­
hour
EbC50
(
area
under
curve)
=
116
(
49
­
142)
mg/
L
72­
hour
ErC50
(
growth
rate)
=
174
(
146
­
208)
mg/
L
72­
hour
EC90
(
cell
density)
=
224
(
193
­
275)
mg/
L
72­
hour
EbC90
(
area
under
curve)
=
204
(
134
­
226)
mg/
L
72­
hour
ErC90
(
growth
rate)
=
275
(
162
­
330)
mg/
L
96­
hour
EC10
(
cell
density)
=
82
(
29
­
123)
mg/
L
96­
hour
EbC10
(
area
under
curve)
=
56
(
26
­
107)
mg/
L
96­
hour
ErC10
(
growth
rate)
=
109
(
84
­
125)
mg/
L
96­
hour
EC50
(
cell
density)
=
131
(
106
­
142)
mg/
L
96­
hour
EbC50
(
area
under
curve)
=
124
(
104
­
138)
mg/
L
96­
hour
ErC50
(
growth
rate)
=
176
(
169
­
181)
mg/
L
96­
hour
EC90
(
cell
density)
=
213
(
203
­
219)
mg/
L
ENV/
JM/
RD(
2002)
17/
FINAL
179
96­
hour
EbC90
(
area
under
curve)
=
209
(
197
­
218)
mg/
L
96­
hour
ErC90
(
growth
rate)
=
225
(
220
­
235)
mg/
L
72­
hour
NOAEC
(
cell
density,
area
under
curve):
37.9
mg/
L
72­
hour
NOAEC
(
growth
rate):
93.8
mg/
L
96­
hour
NOAEC
(
cell
density,
growth
rate):
93.8
mg/
L
96­
hour
NOAEC
(
area
under
curve):
63.9
mg/
L
All
element
values
based
on
mean
measured
concentrations
Statistical
methods:
Cell
densities,
area
under
the
growth
curve
values,
growth
rates
and
percent
inhibition
values
were
calculated
using
"
The
SAS
System
for
Windows",
Release
6.12.
The
EC10,
EC50,
and
EC90
values
and
95%
confidence
limits
were
calculated
by
linear
interpolation
with
treatment
response
and
exposure
concentration
data
using
TOXSTAT
Version
3.5.
Cell
densities,
areas
under
the
growth
curve
and
growth
rates
at
72
and
96
hours
were
evaluated
for
normality
and
homogeneity
of
variances
using
the
Shapiro­
Wilk's
test
and
Levene's
test,
respectively.
Where
the
data
were
normally
distributed
with
equal
variances,
the
treatment
groups
were
compared
to
the
control
using
Dunnett's
test.
In
the
one
instance
where
data
were
not
normally
distributed,
the
non­
parametric
Kruskal­
Wallis
test
was
used.
Results
of
the
statistical
analyses
were
used
to
determine
the
NOAEC
values.

Analytical
Methodology:
Analyses
of
test
solutions
were
performed
at
Wildlife
International
Ltd.
using
high
performance
liquid
chromatography
with
mass
spectrometric
detection
(
HPLC/
MS).
When
determining
the
concentration
of
the
test
substance
in
the
test
solutions,
the
same
and
most
prominent
peak
response
for
perfluorooctanesulfonate
was
used.
No
attempt
was
made
to
quantify
on
the
basis
of
individual
isomeric
components.
The
LOQ
(
limit
of
quantitation)
was
4.80
mg/
L
in
this
study.
The
mean
percent
recovery
of
matrix
fortifications
analyzed
concurrently
during
sample
analysis
was
103%.
Samples
collected
at
test
initiation
had
measured
values
from
100
to
112%
of
nominal.
The
measured
values
for
the
samples
taken
at
72­
hours
were
99.0
­
110%
of
nominal.
The
measured
values
for
the
samples
taken
at
96­
hours
were
99.0
­
109%
of
nominal.
For
the
abiotic
replicate,
the
measured
value
for
the
sample
taken
at
72­
hours
was
103%
of
nominal
and
for
the
sample
taken
at
96­
hours,
107%
of
nominal.

Summary
of
analytical
chemistry
data:
Nominal
Test
Concentration,
mg/
L
Measured
Values
at
0,
72,
and
96­
hours,
Respectively,
mg/
L
Mean
Measured
Concentration,
mg/
L
Percent
of
Nominal
Negative
Control
All
<
LOQ
<
LOQ
­
37.9
37.9,
38.2,
37.6
37.9
100
58.6
65.6,
62.7,
63.4
63.9
109
88.8
94.3,
97.4,
89.8
93.8
106
139
142,
146,
142
143
103
216
230,
238,
236
235
109
331
331,
328,
329
329
99.4
331
(
abiotic)
Not
analyzed,
342,
356
349
105
ENV/
JM/
RD(
2002)
17/
FINAL
180
Biological
observations
after
96­
hours:
Mean
Measured
Concentration,
mg/
L
Mean
Number
of
Cells
per
mL
Percent
Inhibition
via
Density
Percent
Inhibition
via
Area
Under
the
Curve
Percent
Inhibition
via
Growth
Rate
Negative
Control
569,167
­
­
­
37.9
605,000
­
6.3
3.0
­
1.3
63.9
585,833
­
2.9
13
­
0.97
93.8
492,500
13
24*
3.6
143
228,833
60*
66*
22*
235
2,333
100*
99*
100*
329
8,500
99*
100*
96*
*
Indicates
a
significant
difference
from
the
negative
control
using
the
appropriate
statistical
test
(
p
<
0.05)

Control
response:
satisfactory
Observations:
After
96
hours
of
exposure,
there
were
no
signs
of
aggregation
or
adherence
of
the
algae
to
the
flasks
in
the
negative
control
or
any
treatment
group.
In
addition,
there
were
no
noticeable
changes
in
cell
morphology
when
compared
to
the
negative
control.

Reversibility
of
Growth
Inhibition:
Aliquots
of
the
235
and
329
mg/
L
test
solutions
were
diluted
with
algal
medium
and
cultured
for
nine
days
after
the
exposure
phase
of
the
study
concluded.
Based
on
the
increase
in
growth
observed
by
Day
9
of
the
recovery
phase,
the
effect
on
algal
growth
was
algistatic
at
a
concentration
of
235
mg/
L.
However,
no
algal
cells
were
detected
during
the
recovery
phase
in
the
329
mg/
L
treatment,
indicating
that
PFOS
was
algicidal
at
that
concentration.

CONCLUSIONS
The
potassium
perfluorooctanesulfonate
96­
hour
EC50
and
95%
confidence
interval
for
Anabaena
flosaquae
was
determined
using
three
calculation
methods.
By
cell
density,
it
was
131
(
106
 
142)
mg/
L,
by
area
under
the
growth
curve
it
was
124
(
104
 
138)
mg/
L
and
by
growth
rate
176
(
169
 
181)
mg/
L.
The
96­
hour
NOAEC
values
were
determined
to
be
63.9
mg/
L
using
the
area
under
the
growth
curve,
and
93.8
mg/
L
with
the
cell
density
and
growth
rate
calculation
method.
No
signs
of
cell
aggregation
or
adherence
were
noted
in
any
of
the
test
solutions
or
the
controls.
PFOS
was
determined
to
be
algistatic
at
a
concentration
of
235
mg/
L
and
algicidal
at
329
mg/
L.

Submitter:
3M
Company,
Environmental
Laboratory,
P.
O.
Box
33331,
St.
Paul,
Minnesota,
55133
DATA
QUALITY
Reliability:
Klimisch
ranking
=
1
REFERENCES
This
study
was
conducted
at
Wildlife
International
Ltd.,
Easton,
MD
at
the
request
of
the
3M
Company,
Lab
Request
number
U2723.

OTHER
Last
changed:
7/
19/
01
ENV/
JM/
RD(
2002)
17/
FINAL
181
Robust
Study
Report
Reference
No.
37
­
PFOS:
A
7­
day
toxicity
test
with
Duckweed
(
 










G3)

TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
Sample
obtained
from
3M
production
lot
number
217.
The
test
substance
is
a
white
powder.
Sample
was
stored
under
ambient
conditions
prior
to
testing.
Purity
determined
to
be
86.9%
by
LC/
MS,
1H­
HMR,
19F­
NMR
and
elemental
analyses
techniques.

METHOD
Method:
OPPTS
850.4400
Test:
Static
acute
GLP:
Yes
Year
completed:
2001
Species:
Lemna
gibba
G3
Source:
Originally
from
The
United
States
Department
of
Agriculture.
Maintained
in
culture
medium
at
Wildlife
International
Ltd.,
Easton,
MD
Analytical
monitoring:
Test
concentrations
measured
at
0,
3,
5,
and
7­
days
Element
basis:
Number
of
fronds
Exposure
period:
7­
days
Start
date:
3/
3/
00
End
date:
3/
10/
00
Test
organisms
laboratory
culture:
Duckweed
cultures
had
been
actively
growing
in
freshwater
medium
(
20X
AAP)
for
at
least
two
weeks
prior
to
test
initiation.
Stock
nutrient
solutions
were
prepared
by
adding
reagent­
grade
chemicals
to
reverse
osmosis­
purified
well
water.
Test
Conditions:
Test
temperature
range:
24.2
 
25.2
º
C
Light
levels:
5000
+
750
lux
from
continuous
warm­
white
fluorescent
lighting
Growth
medium:
USEPA
OPPTS
850.4400
20X
AAP,
1996
Compound
Nominal
Concentration
Units
MgCl2
.6H20
243.2
mg/
L
CaCl2
.2H20
88.0
mg/
L
H3BO3
3.712
mg/
L
MnCl2
.4H20
8.32
mg/
L
ZnCl2
65.6
µ
g/
L
FeCl3
.6H20
3.196
mg/
L
CoCl2
.6H20
28.56
µ
g/
L
Na2Mo04.2H20
145.2
µ
g/
L
CuCl2
.2H20
0.240
µ
g/
L
Na2EDTA.
2H20
6.00
mg/
L
NaNO3
510
mg/
L
MgSO4
.7H20
294
mg/
L
K2HPO4
20.88
mg/
L
NaHCO3
300
mg/
L
ENV/
JM/
RD(
2002)
17/
FINAL
182
The
pH
of
the
medium
was
adjusted
to
7.5
+
0.1
using
10%
HCl.
Dilution
water
source:
Wildlife
International
Ltd.
well
water
purified
by
reverse
osmosis.
The
test
medium
was
prepared
by
adding
the
appropriate
volumes
of
stock
nutrient
solutions
to
purified
well
water.
The
pH
of
the
medium
was
adjusted
to
7.5
+
0.1
using
10%
HCl
and
the
medium
was
sterilized
by
filtration
(
0.22
µ
m)
prior
to
use.
Stock
and
test
solution
preparation:
A
primary
stock
solution
was
prepared
in
duckweed
medium
at
a
concentration
of
351
mg/
L.
The
primary
stock
solution
was
stirred
with
a
magnetic
stir
plate
for
approximately
24
hours.
After
mixing,
the
primary
stock
solution
was
proportionally
diluted
with
duckweed
medium
to
prepare
the
five
additional
test
concentrations.
All
final
test
solutions
appeared
clear
and
colorless.
Exposure
vessels:
250
mL
plastic
beakers
containing
100
mL
test
solution,
each
covered
with
a
disposable
petri
dish
lid.
Agitation:
None
Number
of
replicates:
three
plus
2
additional
replicates
for
analytical
sampling
on
Days
3
and
5
Initial
loading:
5
plants/
replicate,
15
fronds/
replicate
Number
of
concentrations:
six
plus
a
negative
control
plus
abiotic
controls
at
the
highest
concentration
tested
Water
chemistry:
pH
range
(
0
 
96
hours)
7.9
 
8.9
(
control
exposure)
8.4
 
8.7
(
230
mg/
L
exposure)
Method
of
calculating
mean
measured
concentrations:
arithmetic
mean
obtained
using
results
obtained
at
Days
0,
3,
5,
and
7.

RESULTS
Nominal
concentrations:
Negative
control,
11,
22,
43.9,
87.9,
176,
and
351
mg/
L
plus
351
mg/
L
abiotic
control.
Measured
concentrations:
<
LOQ,
7.74,
15.1,
31.9,
62.5,
147,
230
mg/
L;
abiotic
control
=
231
mg/
L
Element
value
and
95%
confidence
interval
(
based
on
frond
number):
3­
day
IC
10:
101
mg/
L
(
C.
I.
not
calculable)
3­
day
IC
50:
>
230
mg/
L
(
C.
I.
not
calculable)
3­
day
IC
90:
>
230
mg/
L
(
C.
I.
not
calculable)
5­
day
IC
10:
30.7
mg/
L
(
13.3
 
142
mg/
L)
5­
day
IC
50:
182
mg/
L
(
89.1
 
240
mg/
L)
5­
day
IC
90:
>
230
mg/
L
(
C.
I.
not
calculable)
7­
day
IC10:
22.1
mg/
L
(
13.3
 
26.0
mg/
L)
7­
day
IC
50:
108
mg/
L
(
45.7
 
144
mg/
L)
7­
day
IC
90:
>
230
mg/
L
(
C.
I.
not
calculable)
7­
day
NOAEC
(
number
of
fronds):
15.1
mg/
L
All
element
values
based
on
mean
measured
concentrations
Statistical
methods:
Mean
plant
and
frond
numbers,
percent
inhibition
values
and
the
percentages
of
necrotic,
chlorotic
and
dead
fronds
were
calculated
using
"
Microsoft
Excel
Version
5.0",
while
statistical
analyses
were
conducted
using
"
TOXSTAT
Version
3.5".
Percent
inhibition
values
were
calculated
for
each
treatment
group
as
the
percent
reduction
in
mean
frond
number
relative
to
mean
frond
number
in
the
control
replicates.
The
IC
10,
IC50,
and
IC
90
values
and
95%
confidence
intervals
were
determined,
when
possible,
using
linear
interpolation
with
frond
number
and
exposure
concentration
data.
The
percentages
of
dead,
chlorotic
and
necrotic
fronds
also
were
calculated
relative
to
the
total
number
of
fronds
in
each
test
chamber.
The
frond
number
data
was
evaluated
for
normality
and
homogeneity
of
variances
(
p
=
0.05)
ENV/
JM/
RD(
2002)
17/
FINAL
183
using
the
Shapiro­
Wilks'
and
Levene's
tests,
respectively.
The
data
were
normally
distributed
and
the
variances
were
homogeneous,
thus
statistically
significant
differences
between
the
control
and
treatment
groups
were
identified
using
ANOVA
and
Dunnett's
test.
Results
of
the
statistical
analyses,
as
well
as
an
evaluation
of
the
concentration­
response
pattern
and
other
observations
of
effects
were
used
in
the
determination
of
the
no
­
observed­
adverse­
effect­
concentration
(
NOAEC).

Analytical
Methodology:
Analyses
of
test
solutions
were
performed
at
Wildlife
International
Ltd.
using
high
performance
liquid
chromatography
with
mass
spectrometric
detection
(
HPLC/
MS).
Samples
were
centrifuged
as
necessary
prior
to
analysis.
When
determining
the
concentration
of
the
test
substance
in
the
test
solutions,
the
same
and
most
prominent
peak
response
for
perfluorooctanesulfonate
was
used.
No
attempt
was
made
to
quantify
on
the
basis
of
individual
isomeric
components.
The
LOQ
(
limit
of
quantitation)
was
4.39
mg/
L
in
this
study.
The
mean
percent
recovery
of
matrix
fortifications
analyzed
concurrently
during
sample
analysis
was
104%.
Samples
collected
at
test
initiation
had
measured
values
from
64.2
to
82.6%
of
nominal.
Measured
values
for
samples
taken
at
Day
3
ranged
from
67.3
to
83.3%
of
nominal.
Measured
values
for
samples
taken
at
Day
5
ranged
from
65.4
to
85.4%
of
nominal.
Samples
collected
at
test
termination
(
Day
7)
ranged
from
63.9
to
83.8%
of
nominal.
For
the
abiotic
controls,
measured
values
for
samples
taken
at
Day
3,
Day
5,
and
Day
7
ranged
from
64.2
 
66.9%
of
nominal.

Summary
of
analytical
chemistry
data:
Nominal
Test
Concentration,
mg/
L
Measured
Values
at
Days
0,
3,
5,
and
7,
Respectively,
mg/
L
Mean
Measured
Concentration,
mg/
L
Percent
of
Nominal
Negative
Control
All
<
LOQ
<
LOQ
11
7.57,
8.35,
7.47,
7.55
7.74
70
22
15.2,
15.4,
14.6,
15.2
15.1
69
43.9
32.2,
31.9,
31.8,
31.7
31.9
73
87.9
63.5,
63.1,
61.5,
61.8
62.5
71
176
145,
146,
150,
147
147
84
351
226,
237,
232,
224
230
66
351
(
abiotic)
not
analyzed,
225,
235,
232
231
66
Biological
observations
after
7­
Days:
Counts
Mean
Measured
Concentration,
mg/
L
Mean
Number
of
Plants
Mean
Number
of
Fronds
Percent
Inhibition
via
Frond
Number
Negative
Control
19
197
­
7.74
18
177
10
15.1
20
219
­
11
31.9
14
151*
24
62.5
11
134*
32
147
15
69*
65
230
17
37*
81
*
Statistically
significant
difference
(
p
<
0.05)
from
the
negative
control
using
ANOVA
and
Dunnett's
Test.
ENV/
JM/
RD(
2002)
17/
FINAL
184
Effects
Mean
Measured
Concentration,
mg/
L
Mean
Dead
Fronds,
%
Mean
Chlorotic
Fronds,
%
Mean
Necrotic
Fronds,
%
Negative
Control
0
0
0
7.74
0
0
0
15.1
0
1.1
0
31.9
0
0
0.23
62.5
0
0.9
0.61
147
1.0
11
4.5
230
3.8
9.4
19
Control
response:
satisfactory.
Plants
appeared
healthy
and
exhibited
normal
growth
throughout
the
test
with
the
exception
of
one
necrotic
frond
observed
on
Day
3
and
Day
5
of
the
test.

Observations:
Duckweed
exposed
to
147
and
230
mg
PFOS/
L
exhibited
a
dose­
responsive
increase
in
the
incidence
of
dead,
chlorotic
or
necrotic
fronds
during
the
test.
By
Day
7,
all
treatment
groups
>
31.9
gm/
L
showed
evidence
of
sublethal
effects,
including
root
destruction
and/
or
a
cupping
of
the
plant
downward
on
the
water
surface.

CONCLUSIONS
The
potassium
perfluorooctanesulfonate
7­
Day
IC
50
and
95%
confidence
interval
for
duckweed
was
determined
to
be
108
(
45.7
 
144)
mg/
L.
The
7­
Day
NOAEC,
based
on
the
inhibition
of
frond
production
and
evidence
of
sub­
lethal
effects,
was
15.1
mg/
L.

Submitter:
3M
Company,
Environmental
Laboratory,
P.
O.
Box
33331,
St.
Paul,
Minnesota,
55133
DATA
QUALITY
Reliability:
Klimisch
ranking
=
1
REFERENCES
This
study
was
conducted
at
Wildlife
International
Ltd.,
Easton,
MD
at
the
request
of
the
3M
Company.

OTHER
Last
changed:
7/
19/
01
ENV/
JM/
RD(
2002)
17/
FINAL
185
Robust
Study
Report
Reference
No.
38
­
PFOS:
A
96­
hour
toxicity
test
with
freshwater
diatom
(
!





	




	
	



	



)

TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
Sample
obtained
from
3M
production
lot
number
217.
The
test
substance
is
a
white
powder.
Sample
was
stored
under
ambient
conditions
prior
to
testing.
Purity
determined
to
be
86.9%
by
LC/
MS,
1H­
HMR,
19F­
NMR
and
elemental
analyses
techniques.

METHOD
Method:
OPPTS
850.5400
Test:
Acute
static
GLP:
Yes
Year
completed:
2001
Species:
Navicula
pelliculosa
Source:
Originally
from
The
Culture
Collection
of
Algae
at
the
University
of
Texas
at
Austin,
maintained
in
culture
medium
at
Wildlife
International
Ltd.,
Easton,
MD
Analytical
monitoring:
PFOS
measured
at
0,
72,
96­
hours
Element
basis:
Reported
three
ways:
number
of
cells/
ml,
area
under
the
growth
curve
and
growth
rate
Exposure
period:
96­
hours
Start
date:
2/
25/
00
End
date:
2/
29/
00
Test
organisms
laboratory
culture:
Algae
cultures
had
been
actively
growing
in
freshwater
algal
culture
medium
with
silica
and
selenium
for
at
least
two
weeks
prior
to
test
initiation.
Stock
nutrient
solutions
were
prepared
by
adding
reagent­
grade
chemicals
to
reverse
osmosis­
purified
well
water.
Test
Conditions:
Growth
medium
Compound
Nominal
Concentration
Units
MgCl2
.6H20
12.16
mg/
L
CaCl2
.2H20
4.40
mg/
L
H3BO3
0.1856
mg/
L
MnCl2
.4H20
0.416
mg/
L
ZnCl2
3.28
µ
g/
L
FeCl3
.6H20
0.1598
mg/
L
CoCl2
.6H20
1.428
µ
g/
L
Na2Mo04.2H20
7.26
µ
g/
L
CuCl2
.2H20
0.012
µ
g/
L
Na2EDTA.
2H20
0.300
mg/
L
NaNO3
25.50
mg/
L
MgSO4
.7H20
14.70
mg/
L
K2HPO4
1.044
mg/
L
NaHCO3
15.0
mg/
L
Na2SiO3
.9H20
20.0
mg/
L
Na2SeO3
.5H20
0.010
mg/
L
ENV/
JM/
RD(
2002)
17/
FINAL
186
Dilution
water
source:
Wildlife
International
Ltd.
well
water
purified
by
reverse
osmosis.
The
test
medium
was
prepared
by
adding
the
appropriate
volumes
of
stock
nutrient
solutions
to
purified
well
water.
The
pH
of
the
medium
was
adjusted
to
7.5
+
0.1
using
10%
HCl
and
0.1
N
NaOH.
The
medium
was
sterilized
by
filtration
(
0.22
µ
m)
prior
to
use.
Test
solution
preparation:
A
primary
stock
solution
was
not
prepared
for
this
study.
Individual
test
solutions
were
prepared
in
algal
medium
at
each
of
the
seven
nominal
concentrations.
The
individual
test
solutions
were
stirred
with
a
magnetic
stir
plate
for
approximately
24
hours.
All
final
test
solutions
appeared
clear
and
colorless.
Exposure
vessels:
Sterile
250
mL
plastic
Erlenmeyer
flasks
plugged
with
foam
stoppers
containing
100
mL
of
test
solution.
Agitation:
Shaken
continuously
at
~
100
rpm
Number
of
replicates:
three.
Initial
algal
cell
loading:
1.0
X
104
cells/
mL
Number
of
concentrations:
seven
plus
a
negative
control
plus
an
abiotic
control
at
the
highest
concentration
tested
Water
chemistry:
pH
range
(
0
 
96
hours)
7.5
 
8.6
(
control
exposure)
7.5
 
7.7
(
335
mg/
L
exposure)
Test
temperature
range
(
0
 
96
hours)
23.1
 
24.6
º
C
Light
levels:
(
0
 
96
hours)
3910
 
4510
lux
from
continuous
cool­
white
fluorescent
lighting
Method
of
calculating
mean
measured
concentrations:
arithmetic
mean
obtained
using
results
obtained
at
0­
hours,
72­
hours
and
96­
hours
RESULTS
Nominal
concentrations:
Negative
control,
61.5,
81.3,
110,
147,
198,
264,
347
mg/
L
plus
347
mg/
L
abiotic
control.
Measured
concentrations:
<
LOQ,
62.3,
83.2,
111,
150,
206,
266,
335
mg/
L;
abiotic
control
=
339
mg/
L
Element
value
(
95%
confidence
interval):
24­
hour
EC50
(
cell
density)
=
281
(
214
­
312)
mg/
L
24­
hour
EbC50
(
area
under
curve)
=
262
(
205
­
308)
mg/
L
24­
hour
ErC50
(
growth
rate)
=
279
(
212
­
306)
mg/
L
48­
hour
EC50
(
cell
density)
=
261
(
219
­
306)
mg/
L
48­
hour
EbC50
(
area
under
curve)
=
259
(
227
­
303)
mg/
L
48­
hour
ErC50
(
growth
rate)
=
294
(
271
­
307)
mg/
L
72­
hour
EC10
(
cell
density)
=
<
62.3
(
C.
I.
not
calculable)
mg/
L
72­
hour
EbC10
(
area
under
curve)
=
<
62.3
(
C.
I.
not
calculable)
mg/
L
72­
hour
ErC10
(
growth
rate)
=
221
(
190
­
252)
mg/
L
72­
hour
EC50
(
cell
density)
=
242
(
200
­
276)
mg/
L
72­
hour
EbC50
(
area
under
curve)
=
246
(
210
­
277)
mg/
L
72­
hour
ErC50
(
growth
rate)
=
295
(
288
­
305)
mg/
L
72­
hour
EC90
(
cell
density)
=
317
(
306
­
326)
mg/
L
72­
hour
EbC90
(
area
under
curve)
=
318
(
307
­
325)
mg/
L
72­
hour
ErC90
(
growth
rate)
=
335
(
323
­
335)
mg/
L
96­
hour
EC10
(
cell
density)
=
<
62.3
(
C.
I.
not
calculable)
mg/
L
96­
hour
EbC10
(
area
under
curve)
=
<
62.3
(
C.
I.
not
calculable)
mg/
L
96­
hour
ErC10
(
growth
rate)
=
243
(
209
­
295)
mg/
L
96­
hour
EC50
(
cell
density)
=
263
(
217
­
299)
mg/
L
ENV/
JM/
RD(
2002)
17/
FINAL
187
96­
hour
EbC50
(
area
under
curve)
=
252
(
220
­
285)
mg/
L
96­
hour
ErC50
(
growth
rate)
=
305
(
295
­
316)
mg/
L
96­
hour
EC90
(
cell
density)
=
322
(
310
­
328)
mg/
L
96­
hour
EbC90
(
area
under
curve)
=
319
(
308
­
326)
mg/
L
96­
hour
ErC90
(
growth
rate)
=
>
335
mg/
L
(
C.
I.
not
calculable)
72­
hour
NOAEC
(
cell
density,
area
under
the
curve):
<
62.3
mg/
L
72­
hour
NOAEC
(
growth
rate):
206
mg/
L
96­
hour
NOAEC
(
cell
density):
150
mg/
L
96­
hour
NOAEC
(
area
under
the
curve):
<
62.3
mg/
L
96­
hour
NOAEC
(
growth
rate):
206
mg/
L
All
element
values
based
on
mean
measured
concentrations
Statistical
methods:
Cell
densities,
area
under
the
growth
curve
values,
growth
rates
and
percent
inhibition
values
were
calculated
using
"
The
SAS
System
for
Windows",
Release
6.12.
These
values
were
then
analyzed
by
linear
interpolation
using
TOXSTAT
Version
3.5
to
estimate
the
EC10,
EC50,
and
EC90
values
and
95%
confidence
limits.
Cell
densities,
areas
under
the
growth
curve
and
growth
rates
at
72
and
96
hours
were
also
evaluated
fo
r
normality
and
homogeneity
of
variances
using
the
Shapiro­
Wilkes's
test
and
Levene's
test,
respectively.
The
treatment
groups
were
then
compared
to
the
control
using
Dunnett's
test.
Results
of
the
statistical
analyses
were
used
to
determine
the
NOAEC
values.

Analytical
Methodology:
Analyses
of
test
solutions
were
performed
at
Wildlife
International
Ltd.
using
high
performance
liquid
chromatography
with
mass
spectrometric
detection
(
HPLC/
MS).
When
determining
the
concentration
of
the
test
substance
in
the
test
solutions,
the
same
and
most
prominent
peak
response
for
perfluorooctanesulfonate
was
used.
No
attempt
was
made
to
quantify
on
the
basis
of
individual
isomeric
components.
The
LOQ
(
limit
of
quantitation)
was
4.39
mg/
L
in
this
study.
The
mean
percent
recovery
of
matrix
fortifications
analyzed
concurrently
during
sample
analysis
was
108%.
Samples
collected
at
test
initiation
had
measured
values
from
96.2
to
106%
of
nominal.
Measured
values
for
samples
taken
at
72
hours
ranged
from
98.5
to
106%
of
nominal.
Measured
values
for
samples
taken
at
96
hours
ranged
from
94.8
to
101%
of
nominal.
For
the
abiotic
controls,
the
measured
value
for
the
sample
taken
at
72
hours
was
98.2%
of
nominal
and
for
the
sample
taken
at
96
hours,
96.8%
of
nominal.

Summary
of
analytical
chemistry
data:
Nominal
Test
Concentration,
mg/
L
Measured
Values
at
0,
72,
and
96­
hours,
Respectively,
mg/
L
Mean
Measured
Concentration,
mg/
L
Percent
of
Nominal
Negative
Control
All
<
LOQ
<
LOQ
61.5
62.3,
63.6,
61.1
62.3
101
81.3
83.8,
84.7,
81.0
83.2
102
110
109,
113,
110
111
101
147
147,
154,
149
150
102
198
209,
209,
199
206
104
264
271,
268,
258
266
101
347
334,
342,
329
335
96.5
347
(
abiotic)
Not
analyzed,
341,
336
339
97.7
ENV/
JM/
RD(
2002)
17/
FINAL
188
Biological
observations
after
96­
hours:
Mean
Measured
Concentration,
mg/
L
Mean
Number
of
Cells
per
mL
Percent
Inhibition
via
Density
Percent
Inhibition
via
Area
Under
the
Curve
Percent
Inhibition
via
Growth
Rate
Negative
Control
2,726,667
­
­
­
62.3
2,366,667
13
24*
2.5
83.2
2,366,667
13
22*
2.7
111
2,373,333
13
24*
2.5
150
2,473,333
9.3
16
1.7
206
2,093,333
23*
24*
4.7
266
1,330,000
51*
58*
13*
335
35,333
99*
99*
79*

*
Indicates
a
significant
difference
from
the
negative
control
using
Dunnett's
test
(
p
<
0.05)

Control
response:
satisfactory
Observations:
After
96
hours
of
exposure,
there
were
no
signs
of
aggregation
or
adherence
of
the
algae
to
the
flasks
in
the
negative
control
or
any
test
treatment
group.
In
addition,
there
were
no
noticeable
changes
in
cell
color
or
morphology
when
compared
to
the
negative
control,
although
at
72
and
96
hours
of
exposure
a
few
cells
in
the
335
mg/
L
treatment
group
appeared
small
in
comparison
to
the
control.

Reversibility
of
Growth
Inhibition:
The
335
mg/
L
treatment
group
was
maximally
inhibited
after
96­
hours.
The
treatment
group
was
diluted
to
a
concentration
of
the
test
substance
that
would
not
inhibit
growth
and
exposed
for
7
days.
Based
on
the
growth
observed
in
the
recovery
phase,
the
effect
on
algal
growth
was
found
to
be
algistatic.

CONCLUSIONS
The
potassium
perfluorooctanesulfonate
96­
hour
EC50
and
95%
confidence
interval
for
Navicula
pelliculosa
was
determined
using
three
calculation
methods.
B
y
cell
density,
it
was
263
(
217
­
299)
mg/
L,
by
area
under
the
growth
curve
it
was
252
(
220
­
285)
mg/
L
and
by
growth
rate
305
(
295
­
316)
mg/
L.
The
96­
hour
NOAEC
was
determined
by
Dunnett's
procedure
(
p
<
0.05)
to
be
150
mg/
L
using
cell
density,
<
62.3
mg/
L
when
using
area
under
the
curve
and
206
mg/
L
by
growth
rate.
No
signs
of
cell
aggregation
or
adherence
were
noted
in
any
of
the
test
solutions
or
the
controls.
This
test
substance
was
determined
to
be
algistatic.

Submitter:
3M
Company,
Environmental
Laboratory,
P.
O.
Box
33331,
St.
Paul,
Minnesota,
55133
DATA
QUALITY
Reliability:
Klimisch
ranking
=
1
REFERENCES
This
study
was
conducted
at
Wildlife
International
Ltd.,
Easton,
MD
at
the
request
of
the
3M
Company,
Lab
Request
number
U2723.

OTHER
Last
changed:
6/
19/
01
ENV/
JM/
RD(
2002)
17/
FINAL
189
Robust
Study
Report
Reference
No.
39
­
PFOS:
A
96­
hour
toxicity
test
with
the
marine
diatom
(

"

	
















)

TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
Sample
from
3M
production
lot
number
217.
The
test
substance
is
a
white
powder.
Purity
determined
to
be
86.9%
by
LC/
MS,
1H­
HMR,
19F­
NMR
and
elemental
analyses
techniques.

METHOD
Method:
OPPTS
850.5400
Test:
Acute
static
GLP:
Yes
Year
completed:
2001
Species:
Skeletonema
costatum
Source:
Originally
from
The
Culture
Collection
of
Algae
and
Protozoa,
Dunstaffnage
marine
Laboratory
at
Oban
Argyll,
Scotland,
and
maintained
in
culture
medium
at
Wildlife
International
Ltd.,
Easton,
MD
Analytical
monitoring:
PFOS
measured
at
0,
72,
96­
hours
Element
basis:
Reported
three
ways:
number
of
cells/
ml,
area
under
the
growth
curve
and
growth
rate
Exposure
period:
96­
hours
Start
date:
5/
19/
00
End
date:
5/
23/
00
Analytical
monitoring:
Test
concentrations
measured
at
0,
72,
and
96­
hours.
Test
organisms
laboratory
culture:
Algae
cultures
had
been
actively
growing
in
saltwater
algal
culture
medium
for
at
least
two
weeks
prior
to
test
initiation.
Stock
nutrient
solutions
were
prepared
by
adding
reagent­
grade
chemicals
to
reverse
osmosis­
purified
well
water.
Solutions
were
then
diluted
in
artificial
saltwater
to
prepare
final
growth
media.

Test
Conditions:
Algal
saltwater
medium
Compound
Nominal
Concentration
Units
FeCl3
.6H20
0.72
mg/
L
MnCl2
.4H2O
2.16
mg/
L
ZnSO4
.7H2O
0.675
mg/
L
CuSO4
.5H2O
2.36
µ
g/
L
CoCl2
.6H20
6.06
µ
g/
L
H3BO3
17.1
mg/
L
Na2EDTA.
2H20
15.0
mg/
L
K3PO4
3.0
mg/
L
NaNO3
50.0
mg/
L
Na2SiO3
.9H20
20.0
mg/
L
Thiamine
Hydrochloride
0.25
mg/
L
Biotin
0.05
µ
g/
L
B12
0.5
µ
g/
L
ENV/
JM/
RD(
2002)
17/
FINAL
190
Dilution
water
source:
The
stock
nutrient
solutions
were
prepared
by
adding
the
appropriate
volumes
reagent­
grade
chemicals
to
Wildlife
International
Ltd.
well
water
purified
by
reverse
osmosis.
The
algal
medium
was
prepared
by
adding
appropriate
volumes
of
the
stock
nutrient
solutions
to
artificial
saltwater
at
30
ppt
salinity.
The
pH
of
the
medium
was
8.1
and
it
was
sterilized
by
filtration
(
0.22
µ
m)
prior
to
use.
Test
solution
preparation:
A
single
test
solution
(
3.46
mg/
L)
was
prepared
for
this
study
in
algal
saltwater
medium.
The
solution
was
sonicated
for
approximately
30
minutes
and
was
stirred
with
a
magnetic
stir
plate
for
approximately
43
hours.
The
final
test
solution
appeared
clear
and
colorless.
Exposure
vessels:
Sterile
250
mL
glass
Erlenmeyer
flasks
plugged
with
foam
stoppers
containing
100
mL
of
test
solution.
Agitation:
Shaken
continuously
at
100
rpm
Number
of
replicates:
six.
Initial
algal
cell
loading:
7.7
X
104
cells/
mL
Number
of
concentrations:
one
plus
a
negative
control
plus
an
abiotic
control
at
the
highest
concentration
tested
Water
chemistry:
pH
range
(
0
 
96
hours)
8.0
 
8.4
(
control
exposure)
8.0
 
8.4
(
3.20
mg/
L
exposure)
Test
temperature
range
(
0
 
96
hours)
20.2
 
21.4
º
C
Light
levels:
(
0
 
96
hours)
3880
 
4710
lux
from
cool­
white
fluorescent
lighting
Photoperiod:
14­
hours
light
and
10
hours
dark
Method
of
calculating
mean
measured
concentrations:
arithmetic
mean
obtained
using
results
obtained
at
0­
hours,
72­
hours
and
96­
hours
RESULTS
Nominal
concentrations:
Negative
control,
3.46
mg/
L
plus
3.46
mg/
L
abiotic
control.
This
is
apparently
the
highest
concentration
of
PFOS
attainable
in
this
saltwater
algal
media.
Measured
concentrations:
<
LOQ,
3.20
mg/
L;
abiotic
control
=
3.18
mg/
L
Element
value
(
95%
confidence
interval):
72
and
96­
hour
EC10
via
cell
density,
area
under
the
curve
and
growth
rate:
>
3.20
mg/
L
(
C.
I.
not
calculable)
24,
48,
72,
and
96­
hour
EC50
via
cell
density,
area
under
the
curve
and
growth
rate:
>
3.20
mg/
L
(
C.
I.
not
calculable)
72
and
96­
hour
EC90
via
cell
density,
area
under
the
curve
and
growth
rate:
>
3.20
mg/
L
(
C.
I.
not
calculable)
72­
hour
NOAEC
(
cell
density,
area
under
the
curve,
growth
rate):
3.20
mg/
L
96­
hour
NOAEC
(
cell
density,
area
under
the
curve,
growth
rate):
3.20
mg/
L
All
element
values
based
on
mean
measured
concentrations
Statistical
methods:
Cell
densities,
area
under
the
growth
curve
values,
growth
rates
and
percent
inhibition
values
were
calculated
using
"
The
SAS
System
for
Windows",
Release
6.12.
The
EC10,
EC50,
and
EC90
values
and
95%
confidence
limits
could
not
be
calculated
using
statistical
methods.
Cell
densities,
areas
under
the
growth
curve
and
growth
rates
at
72
and
96
hours
were
evaluated
for
normality
using
the
Shapiro­
Wilk's
test
and
for
equality
of
variance
using
an
F­
test.
The
treatment
groups
were
then
compared
to
the
control
using
ANOVA
and
a
2­
sample
t­
test.
ENV/
JM/
RD(
2002)
17/
FINAL
191
Results
of
the
statistical
analyses
were
used
to
determine
the
NOAEC
values.

Analytical
Methodology:
Analyses
of
test
solutions
were
performed
at
Wildlife
International
Ltd.
using
high
performance
liquid
chromatography
with
mass
spectrometric
detection
(
HPLC/
MS).
The
72
and
96­
hour
samples
were
centrifuged
approximately
10
minutes
at
approximately
2000
rpm
prior
to
analysis.
When
determining
the
concentration
of
the
test
substance
in
the
test
solutions,
the
same
and
most
prominent
peak
response
for
perfluorooctanesulfonate
was
used.
No
attempt
was
made
to
quantify
on
the
basis
of
individual
isomeric
components.
The
LOQ
(
limit
of
quantitation)
was
0.480
mg/
L
in
this
study.
The
mean
percent
recovery
of
matrix
fortifications
analyzed
concurrently
during
sample
analysis
was
108%.
Samples
collected
at
test
initiation
had
measured
values
from
96.2
to
88.5%
of
nominal.
The
measured
value
for
the
sample
taken
at
72
hours
was
92.2%
of
nominal.
The
measured
value
for
the
sample
taken
at
96
hours
was
91.2%
of
nominal.
For
the
abiotic
control,
the
measured
value
for
the
sample
taken
at
72
hours
was
97.1%
of
nominal
and
for
the
sample
taken
at
96
hours,
86.8%
of
nominal.

Summary
of
analytical
chemistry
data:
Nominal
Test
Concentration,
mg/
L
Measured
Values
at
0,
72,
and
96­
hours,
Respectively,
mg/
L
Mean
Measured
Concentration,
mg/
L
Percent
of
Nominal
Negative
Control
All
<
LOQ
<
LOQ
3.46
3.26,
3.19,
3.15
3.20
92.5
3.46
(
abiotic)
Not
analyzed,
3.36,
3.00
3.18
91.9
Biological
observations
after
96­
hours:
Mean
Measured
Concentration,
mg/
L
Mean
Number
of
Cells
per
mL
Percent
Inhibition
via
Density
Percent
Inhibition
via
Area
Under
the
Curve
Percent
Inhibition
via
Growth
Rate
Negative
Control
2,481,667
­
­
­
3.20
2,601,667
­
4.8
­
7.3
­
1.3
Control
response:
satisfactory
Observations:
After
96
hours
of
exposure,
there
were
no
signs
of
aggregation
or
adherence
of
the
algae
to
the
flasks
in
the
treatment
group.
However
there
were
signs
of
adherence
to
the
test
chamber
in
the
negative
control
group.
There
were
no
noticeable
changes
in
cell
morphology
when
compared
to
the
negative
control.

Reversibility
of
Growth
Inhibition:
After
96­
hours
of
exposure,
there
was
no
significant
inhibition
of
growth
in
the
highest
concentration
tested
(
3.20
mg/
L).
Therefore,
a
recovery
phase
was
not
conducted.

CONCLUSIONS
A
single
concentration
of
potassium
perfluorooctanesulfonate
was
evaluated
for
toxicity
to
Skeletonema
costatum.
This
mean
measured
concentration,
3.20
mg/
L,
was
the
highest
concentration
attainable
in
this
algal
media.
The
96­
hour
EC50
and
95%
confidence
interval
for
Skeletonema
costatum,
as
determined
by
cell
density,
area
under
the
growth
curve,
and
by
growth
rate
was
found
to
be
>
3.20
mg/
L.
The
96­
hour
NOAEC
was
determined
by
ANOVA
and
a
2­
sample
t­
test
to
be
3.20
mg/
L
calculated
using
cell
density,
area
under
the
curve
and
growth
rate.
No
signs
of
cell
aggregation
or
adherence
were
noted
in
any
of
the
test
solutions
or
the
controls.
ENV/
JM/
RD(
2002)
17/
FINAL
192
Submitter:
3M
Company,
Environmental
Laboratory,
P.
O.
Box
33331,
St.
Paul,
Minnesota,
55133
DATA
QUALITY
Reliability:
Klimisch
ranking
=
1
REFERENCES
This
study
was
conducted
at
Wildlife
International
Ltd.,
Easton,
MD
at
the
request
of
the
3M
Company,
Lab
Request
number
U2723.

OTHER
Last
changed:
7/
19/
01
ENV/
JM/
RD(
2002)
17/
FINAL
193
Robust
Study
Report
Reference
No.
40
­
PFOS:
A
frog
embryo
teratogenesis
assay
 







(
FETAX)

TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS,
U2723
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
Sample
obtained
from
3M
production
lot
number
217.
The
test
substance
is
a
white
powder.
Purity
determined
to
be
86.9%
by
LC/
MS,
1H­
HMR,
19F­
NMR
and
elemental
analyses
techniques.

METHOD
Method:
ASTM
E1439­
91
Test
type:
Static
renewal
GLP:
In­
life
phase
 
no;
stock
solution
preparation
and
measurement
of
test
concentrations
­
yes
Year
completed:
2001
Number
of
studies:
3
Study
1
Study
2
Study
3
Start
date:
5/
15/
00
5/
22/
00
5/
22/
00
End
date:
5/
19/
00
5/
26/
00
5/
26/
00
(
Study
2
and
3
set
up
concurrently
with
common
stock
solutions)
Analytical
monitoring:
PFOS
measured
at
0
and
96­
hours
Species:
Xenopus
laevis
Source:
Breeding
colonies
at
the
University
of
Maryland
Wye
Research
and
Education
Center
(
UMD/
WREC),
Queenstown,
Maryland.
Test
organisms
laboratory
culture:
Mating
pairs
were
bred
in
the
dark
in
23.5
+
0.5
º
C
UMD/
WREC
non­
chlorinated
well
water
at
~
70
day
intervals
by
injecting
400
and
800
I.
U.
of
human
chorionic
gonadotropin
(
HCG)
in
the
dorsal
lymph
sac
of
the
males
and
females,
respectively.
Amplexus
occurred
4
­
6
hours
after
injecting
HCG;
egg
deposition
occurred
9­
12
hours
following
HCG
injection.
Age
at
test
initiation:
Embryos;
normal
stage
8
blastula
to
normal
stage
11
gastrula
Loading:
25
embryos/
10
mL
Pretreatment:
Embryos
de­
jelled
in
a
2%
L
­
cysteine
solution,
then
rinsed
and
re­
suspended
in
FETAX
solution
prior
to
introduction
to
test
chambers.
Element
basis:
mortality,
malformations
(
via
the
atlas
of
Bantle
et
al.,
1991),
growth
Exposure
period:
96­
hours
Test
Conditions
(
all
3
studies):
Dilution
water:
ASTM
(
1998)
FETAX
solution
Test
temperature:
24.0
+
0.2
º
C
Light
levels:
60­
85
foot
candle
fluorescent
lights
Photoperiod:
12­
hour
light:
12­
hour
dark
Stock
and
test
solution
preparation:
A
primary
stock
solution
was
prepared
in
FETAX
medium
(
supplied
by
UM­
WREC)
by
Wildlife
International,
Ltd.
at
48
mg
PFOS/
L.
The
primary
stock
solution
was
mixed
by
sonication
and
stirring.
After
mixing,
the
primary
stock
solution
was
proportionally
diluted
with
FETAX
medium
to
prepare
the
six
test
concentrations.
The
six
test
concentration
solutions
were
delivered
to
UM­
WREC
prior
to
the
start
of
each
study.
Reference
substance:
6­
aminonicotinamide
Stock
and
reference
substance
solution
preparation:
as
outlined
in
the
ASTM
(
1998)
protocol
Exposure
vessels:
Covered
60
mm
glass
Petri
dishes
containing
10
mL
test
solution
ENV/
JM/
RD(
2002)
17/
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194
Number
of
replicates:
controls
 
4,
treatments
 
2
Number
of
embryos
per
replicate:
25
Number
of
concentrations:
six
plus
a
negative
control
plus
an
abiotic
control
at
the
highest
concentration
tested,
plus
two
reference
substance
concentrations.
Renewal
frequency:
every
24
hours
Stability
of
the
test
chemical
solutions:
Extremely
stable
Water
chemistry
during
all
3
studies:
pH
range
(
0
 
96
hours)
7.1
 
7.7
(
control
exposure)
7.0
 
7.6
(
24
mg/
L
nominal
exposure)
Dissolved
oxygen
range
(
0­
96
hours)
7.3
 
8.4
mg/
L
(
control
exposure)
7.0
 
8.5
mg/
L
(
24
mg/
L
nominal
exposure)
Method
of
calculating
mean
measured
concentrations:
arithmetic
mean
RESULTS
Nominal
PFOS
concentrations:
Negative
control,
1.82,
3.07,
5.19,
8.64,
14.4
and
24.0
mg/
L
plus
24.0
mg/
L
abiotic
control.
Nominal
6­
aminonicotinamide
concentrations:
5.5
and
2500
mg/
L
Mean
measured
PFOS
concentrations:
Study
1:
<
LOQ,
2.00,
2.83,
4.73,
7.90,
14.7,
24.6
mg/
L;
abiotic
control
=
23.7
mg/
L
Study
2:
<
LOQ,
1.91,
3.04,
4.82,
7.97,
13.3,
23.1
mg/
L;
abiotic
control
=
23.9
mg/
L
Study
3:
<
LOQ,
1.93,
3.27,
5.25,
8.26,
14.0,
23.9
mg/
L;
abiotic
control
=
24.1
mg/
L
PFOS
element
values
and
95%
confidence
intervals,
mg/
L
Study
Number
96­
Hr
LC50
96­
Hr
EC50
Minimum
conc.
to
Inhibit
Growth
(
MCIG)
Teratogenic
Index
(
TI)

1
13.8
(
12.4
­
15.3)
12.1
(
10.0
 
14.6)
Not
calculable
1.1
2
17.6
(
15.5
 
20.0)
17.6
(
13.5
 
22.9)
7.97
1.0
3
15.3
(
13.1
 
17.8)
16.8
(
12.4
 
22.8)
8.26
0.9
All
element
values
based
on
mean
measured
concentrations
Statistical
methods:
The
Trimmed
Spearman­
Karber
statistical
procedure
was
used
to
determine
the
96­
hour
LC50
for
mortality
and
96­
hour
EC50
for
malformations.
The
MCIG
was
determined
by
Bonferroni's
T­
Test.
All
statistical
tests
were
performed
using
Toxstat
(
WEST
and
Gulley,
1994).
A
minimum
probability
level
of
0.05
was
used.
The
teratogenic
index
(
TI)
was
calculated
by
dividing
the
LC50
by
the
EC50.

Analytical
Methodology:
Analyses
of
test
solutions
were
performed
at
Wildlife
International
Ltd.,
Easton,
MD
using
high
performance
liquid
chromatography
with
mass
spectrometric
detection
(
HPLC/
MS).
When
determining
the
concentration
of
the
test
substance
in
the
test
solutions,
the
same
and
most
prominent
peak
response
for
perfluorooctanesulfonate
was
used.
No
attempt
was
made
to
quantify
on
the
ENV/
JM/
RD(
2002)
17/
FINAL
195
basis
of
individual
isomeric
components.
The
LOQ
(
limit
of
quantitation)
was
0.240
mg/
L
in
these
studies.
The
mean
percent
recovery
of
matrix
fortifications
analyzed
concurrently
during
sample
analysis
was
97.2%.
Samples
collected
at
test
initiation
had
measured
values
from
112
to
141%
of
nominal
in
the
first
study,
and
in
the
second
and
third
studies,
from
95.8
to
117%
of
nominal.
Measured
values
for
samples
taken
at
96­
hours
ranged
from
54.7
to
98.6%
of
nominal
in
the
first
study
and
80.7
to
112%
of
nominal
in
the
second
and
third
studies.
The
samples
from
the
abiotic
24.0
mg/
L
treatment
group
was
comparable
to
samples
from
the
24.0
mg/
L
treatment
group
with
the
embryos
present.

Summary
of
analytical
chemistry
data:
Study
1
Nominal
Test
Concentration,
mg/
L
Measured
Values
at
0
and
96­
hours
Respectively,
mg/
L
Mean
Measured
Concentration,
mg/
L
Percent
of
Nominal
Negative
Control
All
<
LOQ
<
LOQ
1.82
2.58,
1.42
2.00
110
3.07
3.94,
1.72
2.83
92.2
5.19
6.62,
2.84
4.73
91.1
8.64
10.7,
5.09
7.90
91.4
14.4
18.5,
10.8
14.7
102
24.0
26.9,
22.3
24.6
103
24.0
(
abiotic)
not
analyzed,
23.7
23.7
98.6
Study
2
Nominal
Test
Concentration,
mg/
L
Measured
Values
at
0
and
96­
hours
Respectively,
mg/
L
Mean
Measured
Concentration,
mg/
L
Percent
of
Nominal
Negative
Control
All
<
LOQ
<
LOQ
1.82
1.77,
2.04
1.91
105
3.07
3.59,
2.49
3.04
99.0
5.19
5.45,
4.18
4.82
92.9
8.64
8.43,
7.51
7.97
92.2
14.4
14.5,
12.1
13.3
92.4
24.0
23.0,
23.1
23.1
96.3
24.0
(
abiotic)
not
analyzed,
23.9
23.9
99.6
Study
3
Nominal
Test
Concentration,
mg/
L
Measured
Values
at
0
and
96­
hours
Respectively,
mg/
L
Mean
Measured
Concentration,
mg/
L
Percent
of
Nominal
Negative
Control
All
<
LOQ
<
LOQ
1.82
1.77,
2.08
1.93
106
3.07
3.59,
2.94
3.27
107
5.19
5.45,
5.05
5.25
101
8.64
8.43,
8.09
8.26
95.6
14.4
14.5,
13.5
14.0
97.2
24.0
23.0,
24.7
23.9
99.6
24.0
(
abiotic)
not
analyzed,
24.1
24.1
100
ENV/
JM/
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2002)
17/
FINAL
196
Biological
observations
after
96­
hours:

Mortality
and
Malformations
Test
1
Test
2
Test
3
Nominal
Concentration*,
mg/
L
Percent
Mortality
Percent
Malformations
Percent
Mortality
Percent
Malformations
Percent
Mortality
Percent
Malformations
Negative
Control
1.0
4.0
1.0
4.0
0
2.0
1.82
2.0
8.2
0
8.0
0
6.0
3.07
4.0
15
10
4.4
0
4.0
5.19
10
22
8.0
11
0
6.0
8.64
12
25
10
20
14
14
14.4
38
65
30
37
44
39
24.0
100
­
70
67
78
73
*
Nominal
concentrations
used
for
ease
of
comparison
table
Malformations:
The
most
common
types
of
malformations
noted
were
improper
gut
coiling,
edema,
notochord
abnormalities
and
facial
abnormalities.

Growth
 
Mean
length
(
mm)
after
96­
hours
Exposure
Nominal
Concentration*,
mg/
L
Test
1
Test
2
Test
3
Negative
Control
8.59
8.88
9.47
1.82
8.29
8.45
9.10
3.07
8.80
8.57
9.28
5.19
8.51
8.72
9.28
8.64
8.71
7.93**
8.51**
14.4
8.08
7.51**
8.11**
24.0
­
(
total
mortality)
7.39**
7.80**
*
Nominal
concentrations
used
for
ease
of
comparison
table
**
Significantly
different
at
alpha
=
0.05
(
Bonferroni
T­
Test)

Control
response:
satisfactory.

Reference
substance
response:
satisfactory
at
low
concentration
(
5.5
mg/
L).
Did
not
meet
ASTM
(
1998)
criteria
for
high
concentration
(
2,500
mg/
L).
However,
results
obtained
at
high
concentration
were
consistent
and
not
at
variance
with
previous
experience
in
this
testing
laboratory.

Observations:
Majority
of
embryo
mortality
appeared
to
be
caused
by
the
gut
coiling
through
the
body
wall
at
the
two
highest
test
concentrations.

CONCLUSIONS
The
potassium
perfluorooctanesulfonate
96­
hour
LC50
range
for
FETAX
was
determined
to
be
13.8
 
17.6
mg/
L.
The
96­
hour
EC50
range
was
12.1
 
17.6
mg/
L.
The
range
for
Minimum
Concentration
to
Inhibit
Growth
(
MCIG)
was
7.97
to
>
14.7
mg/
L.
The
Teratogenic
Index
(
TI)
was
found
to
be
0.9
 
1.1.
This
TI
range
indicates
that
potassium
perfluorooctanesulfonate
has
a
low
potential
to
be
a
developmental
hazard.
ENV/
JM/
RD(
2002)
17/
FINAL
197
Submitter:
3M
Company,
Environmental
Laboratory,
P.
O.
Box
33331,
St.
Paul,
Minnesota,
55133
DATA
QUALITY
Reliability:
Klimisch
ranking
=
2
Although
these
were
well­
conducted
studies,
the
in­
life
phases
were
not
conducted
in
accordance
with
Good
Laboratory
Practices.

REFERENCES
These
studies
were
conducted
at
the
University
of
Maryland
Wye
Research
and
Education
Center
(
UMWREC
in
Queenstown,
Maryland
and
at
Wildlife
International
Ltd.,
Easton,
MD
at
the
request
of
the
3M
Company.
Lab
Request
number
U2723
OTHER
Last
changed:
7/
19/
01
ENV/
JM/
RD(
2002)
17/
FINAL
198
Robust
Study
Report
Reference
No.
41
­
Perfluorooctanesulfonate,
Potassium
salt
(
PFOS):
A
flowthrough
bioconcentration
test
with
the
Bluegill
(
 

















)

TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
Sample
from
3M
production
lot
number
217.
The
test
substance
is
a
white
powder.
Purity
determined
to
be
86.9%
by
LC/
MS,
1H­
HMR,
19F­
NMR
and
elemental
analyses
techniques.

METHOD:

Method/
guideline
followed:
US
EPA
OPPTS
850.1730
and
OECD
305
Type:
Flow­
through
exposure
with
flow­
through
depuration
phase.
GLP
(
Y/
N):
Yes
Year:
2001
Species:
Bluegill
(
Lepomis
macrochirus)
Supplier:
Osage
Catfisheries,
Inc.,
Osage
Beach,
Missouri
Length
and
weight
at
test
termination:
Mean
length
=
62
mm,
range
56­
66
mm
Mean
weight
=
2.70
g,
range
2.03
 
3.32
g
Loading:
0.48
g
fish/
L/
day
(
based
on
initial
loading
of
90
fish
per
tank,
using
mean
fish
weight
at
the
end
of
the
study
and
volume
of
water
that
passed
through
test
chamber
in
24­
hours).
Fish
age:
Approximately
7
months
at
test
initiation
Analytical
monitoring:
Concentration
of
PFOS
in
water
and
fish.
Pretreatment:
None
Number
of
concentrations:
Two
plus
a
negative
control
Test
concentrations
(
mean
measured):
Negative
control,
0.086
and
0.87
mg/
L
Uptake
period:
62­
days
(
0.086
mg/
L
exposure)
35­
days
(
0.87
mg/
L
exposure
 
this
exposure
ended
after
35­
days
due
to
fish
mortality)
Depuration
period:
56­
days
(
0.086
mg/
L
exposure
None
(
0.087
mg/
L
exposure)
Test
conditions:
Dilution
water:
Moderately
 
hard
well
water
Dilution
water
chemistry:
Specific
conductance:
313
(
310
 
315
µ
mhos/
cm)
Hardness:
130
(
128
 
132
mg/
L)
Alkalinity:
178
(
176
 
178)
pH:
8.1
(
8.0
 
8.2)
Measured
during
the
4
­
week
period
immediately
preceding
the
test.

Stock
and
test
solution
preparation:
Two
stock
solutions
were
prepared
at
10
and
100
mg
a.
i./
L.
Stock
solutions
stirred
with
an
electric
top­
down
mixer
to
aid
in
the
solubilization
of
the
test
substance.
After
mixing,
the
stocks
appeared
clear
and
colorless.
Stocks
were
prepared
at
approximately
weekly
intervals
during
the
uptake
phase.
Stocks
injected
into
the
diluter
mixing
chambers
at
a
rate
of
3.5
mL/
minute
where
they
were
mixed
with
dilution
water
at
a
rate
of
350
mL/
minute
to
achieve
the
desired
test
concentrations.
All
final
test
solutions
appeared
clear
and
colorless.
Diluter
flow
rate:
Approx.
6.3
volume
additions
per
24­
hours
ENV/
JM/
RD(
2002)
17/
FINAL
199
Exposure
vessels:
104
L
stainless
steel
aquaria
filled
with
approximately
80
L
solution.
Number
of
replicates:
None
 
one
vessel
per
concentration
Number
of
fish
per
vessel:
90
Diet:
Flake
food,
Ziegler
Brothers,
Inc.,
Gardners,
PA
Water
chemistry
ranges
during
the
study:
Neg.
Control
0.086
mg/
L
0.87
mg/
L
Dissolved
oxygen,
mg/
L:
6.8
 
8.6
6.8
 
8.6
6.4
 
8.2
Temperature,
°
C:
21.8
 
22.0
21.7
 
22.0
21.7
 
21.9
pH:
7.9
 
8.2
7.9
 
8.2
7.9
 
8.2
Photoperiod:
16
hours
light
and
8
hours
dark
with
a
30­
minute
transition
period.
Light
intensity:
278
lux
at
surface
of
the
negative
control
vessel
at
test
initiation
Collection
of
tissue
samples:
Fish
were
collected
from
test
chambers
by
random
selection
at
12
time
points
during
the
62­
day
uptake
phase.
They
were
euthanized,
blotted
dry,
weighed
and
measured.
Fish
then
rinsed
with
dilution
water,
blotted
dry
again
and
dissected
into
edible
and
non­
edible
tissue
fractions.
The
fractions
were
individually
weighed.
The
head,
fins
and
viscera
were
considered
to
be
non­
edible
tissue.
The
remaining
tissue,
including
skin
was
considered
to
be
edible
tissue.

Statistical
methods:
Whole
fish
concentrations
were
calculated
based
on
the
sum
of
the
edible
and
nonedible
parts.
Steady­
state
bioconcentration
BCF
values
calculated
from
the
tissue
concentrations
at
apparent
steady­
state
divided
by
the
mean
water
concentration.
Tissue
concentrations
were
considered
to
be
at
apparent
steady­
state
if
3
or
more
consecutive
sets
of
tissue
concentrations
were
not
significantly
different
(
p
>
0.05).
Tissue
concentrations
were
evaluated
for
normality
and
homogeneity
of
variance
suing
the
Shapiro­
Wilk's
test
and
Bartlett's
test,
respectively.
If
the
data
did
not
meet
the
assumptions,
data
was
transformed
in
an
attempt
to
correct
the
data.
Mean
tissue
concentrations
were
then
compared
using
ANOVA
and
Dunnett's
test.

The
kinetic
bioconcentration
factor
(
BCFK),
uptake
rate
(
k1)
and
depuration
rate
(
k2)
were
calculated
for
the
edible,
nonedible
and
whole
fish
exposed
to
0.086
mg/
L
PFOS
using
BIOFAC
computer
software.
BIOFAC
is
a
nonlinear
parameter
estimate
routine
which
estimates
rate
constants
from
a
set
of
sequential
time­
concentration
data.
These
rate
constants
were
then
used
to
calculate
a
BCFK
(
BCFK
=
K
/
K
1
2).

RESULTS
Nominal
concentrations:
Negative
control,
0.1
and
1.0
mg/
L
Mean
measured
concentrations:
<
0.05,
0.086
and
0.87
mg/
L
Bioconcentration
factors
(
BCF):

0.086
mg/
L
apparent
steady­
state
BCF
Edible
Non­
edible
Whole
Fish
484
1124
856
0.87
mg/
L
(
study
ended
prior
to
achieving
steady­
state)
BCF:
Edible
Non­
edible
Whole
Fish
136
386
278
ENV/
JM/
RD(
2002)
17/
FINAL
200
BIOFAC
Estimates
(
using
0.086
mg/
L
exposure)
Edible
Non­
edible
Whole
Fish
BCFK:
1866
4312
3614
Time
to
reach
50%
clearance:
146
days
133
days
152
days
PFOS
Concentrations
in
Tissues
of
Bluegill
Exposed
to
0.086
mg/
L
Values
are
from
4
individual
fish
at
each
sample
period.
Uptake
Day
Edible
Tissue,
mg/
kg
Non­
edible
Tissue,
mg/
kg
Whole
Fish
Conc.,
mg/
kg
0
(
4­
hours)
0.167,
0.155,
0.144,
0.182
0.415,
0.519,
0.417,
0.497
0.293,
0.351,
0.286,
0.363
1
0.734,
0.726,
0.631,
0.806
1.68,
1.85,
1.72,
2.07
1.26,
1.34,
1.29,
1.53
3
1.73,
2.07,
2.03,
2.11
4.59,
5.50,
5.47,
5.97
3.21,
4.04,
4.18,
4.38
7
3.73,
4.25,
4.73,
6.25
10.2,
10.6,
11.9,
15.2
7.33,
7.66,
8.73,
11.4
14
11.4,
9.07,
13.7,
12.6
27.3,
23.2,
35.3,
32.6
20.2,
16.9,
26.0,
24.6
21
11.7,
12.0,
12.9,
10.6
33.3,
22.7,
24.6,
24.4
23.3,
18.4,
19.8,
18.5
28
18.3,
13.7,
23.9,
23.1
49.4,
40.7,
65.3,
57.9
35.3,
29.2,
45.4,
44.1
35
22.6,
27.7,
23.8,
20.6
67.1,
73.3,
62.0,
59.1
46.3,
53.8,
46.6,
40.9
42
27.6,
25.3,
21.2,
27.6
64.0,
68.1,
54.4,
79.6
50.1,
49.4,
40.9,
56.3
49
33.3,
36.2,
39.0,
30.6
85.0,
95.1,
93.1,
77.7
62.8,
69.6,
70.8,
57.4
56
48.3,
38.9,
44.1,
38.3
122,
94.2,
73.2,
106
90.6,
71.6,
63.3,
74.8
62
42.4,
66.2,
42.2,
39.2
101,
112,
105,
96.4
77.0,
92.7,
79.6,
73.1
Depuration
Day
14
48.5,
31.8,
31.6,
42.0
124,
79.4,
81.8,
113
90.3,
60.4,
61.6,
85.3
28
26.0,
33.3,
38.7,
55.8
85.7,
95.1,
85.7,
94.8
58.2,
70.1,
68.1,
81.1
42
24.1,
31.2,
30.0,
33.0
71.7,
80.6,
78.3,
82.1
51.4,
61.4,
61.0,
62.2
56
21.1,
37.6,
32.9,
31.2
57.7,
80.3,
85.4,
84.4
41.6,
66.5,
65.8,
62.1
PFOS
Concentrations
in
Tissues
of
Bluegill
Exposed
to
0.87
mg/
L
Values
are
from
4
individual
fish
at
each
sample
period.
Uptake
Day
Edible
Tissue,
mg/
kg
Non­
edible
Tissue,
mg/
kg
Whole
Fish
Conc.,
mg/
kg
0
(
4­
hours)
1.46,
1.48,
1.19,
1.39
3.52,
4.37,
4.22,
4.06
2.71,
3.08,
2.84,
2.89
1
4.68,
6.59,
5.56,
5.64
11.1,
14.2,
13.3,
12.1
8.00,
10.9,
10.2,
9.47
3
17.3,
15.8,
19.0,
20.8
39.3,
42.0,
43.8,
51.8
30.5,
30.7,
34.5,
39.1
7
42.0,
44.0,
57.7,
46.8
100,
102,
102,
120
74.9,
77.0,
85.3,
89.8
14
87.1,
81.6,
90.7,
73.3
177,
207,
245,
214
141,
157,
180,
158
21
79.4,
117,
104,
102
201,
278,
246,
229
146,
210,
185,
172
28(
1)
102,
131,
107,
133
289,
372,
320,
361
205,
267,
232,
263
(
1)
Sampling
of
fish
stopped
after
Uptake
Day
28
due
to
mortality.

Test
organism
mortality:
Negative
control:
None
during
the
uptake
phase
(
62
days)
or
depuration
phase
(
35
days)
0.086
mg/
L
exposure:
One
fish
died
after
49
days
and
one
after
59
days
of
exposure
in
the
uptake
phase,
none
during
the
depuration
phase.
0.87
mg/
L
exposure:
Mortality
first
noted
on
Day
9
and
continued
through
Day
35
of
the
uptake
phase
at
which
time
all
of
the
fish
had
either
died
or
had
been
sampled
Analytical
methodology:
Analyses
of
test
solutions
and
fish
tissues
were
performed
at
Wildlife
ENV/
JM/
RD(
2002)
17/
FINAL
201
International,
Ltd.
Water
samples
were
diluted
and
analyzed
by
HPLC
with
single
quadrupole
mass
spectrometric
detection.
Tissue
samples
were
homogenized,
extracted,
diluted
and
analyzed
by
HPLC
with
triple
quadrupole
mass
spectrometric
detection.
When
determining
the
concentration
of
the
test
substance
in
the
samples,
the
same
and
most
prominent
peak
response
for
perfluorooctanesulfonate
was
used.
No
attempt
was
made
to
quantify
on
the
basis
of
individual
isomeric
components.
The
LOQ
was
0.05
mg/
L
for
water
in
this
study.
For
tissue
samples,
the
LOQ
was
calculated
on
an
individual
basis
for
each
sample
since
each
entire
submitted
sample,
of
differing
weight,
was
extracted
without
an
adjustment
to
constant
weight.

Recovery
was
excellent
in
both
water
and
fish
tissues,
ranging
from
84.9
to
122%
of
fortification
levels.
Analytical
results
were
not
corrected
for
procedural
recovery.

CONCLUSIONS
PFOS
bioconcentrated
in
the
tissues
of
bluegill
sunfish
during
this
study.
Apparent
steady­
state
was
attained
on
Day
49
for
the
fish
exposed
to
0.086
mg
a.
i./
L.
Although
Day
49,
56
and
62
tissue
residues
were
not
statistically
significantly
different,
PFOS
concentrations
appeared
to
be
still
increasing
during
this
time.
Apparent
steady­
state
BCF
values
for
edible,
non­
edible
and
whole
fish
tissues
were
calculated
to
be
484,
1124,
and
859,
respectively.

PFOS
depurated
slowly.
The
BIOFAC
estimates
for
the
time
to
reach
50%
clearance
for
edible,
nonedible
and
whole
fish
tissues
were
146,
133
and
152
days,
respectively.

DATA
QUALITY
Reliability:
Klimisch
ranking
=
1
REFERENCES
This
study
was
conducted
at
Wildlife
International,
Ltd.,
Easton,
MD
at
the
request
of
the
3M
Company,
Lab
Request
number
U2723.

OTHER
Last
changed:
7/
19/
01
ENV/
JM/
RD(
2002)
17/
FINAL
202
Robust
Study
Report
Reference
No.
42
­
Perfluorooctanesulfonate,
Potassium
salt
(
PFOS):
96­
Hour
Static
Acute
Toxicity
Test
with
the
Rainbow
Trout
(
#














"



)
in
freshwater
TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)
Remarks:
The
test
substance
is
a
white
powder.
Sample
was
taken
from
3M
lot
number
217.
Sample
was
stored
under
ambient
conditions
prior
to
testing.
Purity
determined
to
be
86.9%
by
LC/
MS,
1
H­
HMR,
19
F­
NMR
and
elemental
analyses
techniques.

METHOD
Method:
OPPTS
850.1075
and
OECD
203
Type:
Static
acute
GLP:
Yes
Date
completed:
Study
completed
2001,
report
completed
2002
Species:
Oncorhynchus
mykiss
Supplier:
Thomas
Fish
Company,
Anderson,
CA
Analytical
monitoring:
PFOS
measured
at
0,
48,
96­
hours
Exposure
period:
96­
hours
Statistical
methods:
LC50
values
calculated,
when
possible,
by
probit
analysis,
moving
average
method
or
binomial
probability
with
non­
linear
interpolation
using
the
computer
software
of
C.
E.
Stephan.
Test
fish
age:
juveniles
Average
Total
Length
and
weight:
3.6
(
3.4
 
4.0)
cm,
0.34
(
0.25­
0.47)
g
Loading:
0.23
g
fish/
L
Pretreatment:
None
Test
conditions:
Dilution
water:
0.45
µ
m
filtered
moderately
hard
well
water
Dilution
water
chemistry
(
during
the
4­
week
period
immediately
preceding
the
test):
hardness:
130
(
128­
132)
mg/
L
as
CaCO3
alkalinity:
177
(
176­
178)
mg/
L
as
CaCO3
pH:
8.3
(
8.2
 
8.4)
TOC:
Not
given
Conductivity:
311
(
310­
315)
µ
mhos/
cm
Stock
and
test
solution
preparation:
Primary
stock
prepared
in
dilution
water
at
150
mg/
L
and
mixed
for
~
23
hours
prior
to
use.
After
mixing,
primary
stock
solution
was
proportionally
diluted
with
dilution
water
to
prepare
the
five
test
concentrations.

Concentrations
dosing
rate:
Once
Stability
of
the
test
chemical
solutions:
Extremely
stable
Exposure
vessels:
25L
polyethylene
aquaria
containing
approximately
15L
of
test
solution;
water
depth
approx
17.5
cm.
Number
of
replicates:
two
Number
of
fish
per
replicate:
ten
ENV/
JM/
RD(
2002)
17/
FINAL
203
Number
of
concentrations:
five
plus
a
negative
control
Water
chemistry
during
the
study:
Dissolved
oxygen
range
(
0
 
96
hours)
:
9.4
 
10.7
mg/
L
(
control
exposure)
9.2
 
10.8
mg/
L
(
50
mg/
L
exposure)
pH
range
(
0
 
96
hours)
8.1
 
8.4
(
control
exposure)
8.2
 
8.4
(
50
mg/
L
exposure)
Test
temperature
range
(
0
 
96
hours)
12.1
 
12.6
º
C
(
control
exposure)
11.8
 
12.9
º
C
(
50
mg/
L
exposure)
­
Method
of
calculating
mean
measured
concentrations:
arithmetic
mean
RESULTS
Nominal
concentrations:
Bk
control,
3.1,
6.3,
13,
25,
50
(
exposure),
50
(
abiotic)
mg/
L
Measured
concentrations:
<
LOQ,
3.0,
6.3,
13,
25,
50,
52
mg/
L
Element
value:
24­
hour
LC50
=
>
50
mg/
L
(
C.
I.
not
calculable)
48­
hour
LC50
=
>
50
mg/
L
(
C.
I.
not
calculable)
72­
hour
LC50
=
>
50
mg/
L
(
C.
I.
not
calculable)
96­
hour
LC50
=
22
(
18
­
27)
mg/
L
All
element
values
based
on
mean
measured
concentrations
Statistical
Evaluation
of
Mortality:
Element
values
and
confidence
limits
for
24,
48,
and
72­
hours
could
not
be
calculated
due
to
lack
of
mortality.
Probit
Analysis
was
used
to
calculate
the
96­
hour
LC50.

Analytical
Methodology:
Analyses
of
test
solutions
were
performed
at
Wildlife
International
Ltd.
using
high
performance
liquid
chromatography
with
mass
spectrometric
detection
(
HPLC/
MS).
When
determining
the
concentration
of
the
test
substance
in
the
test
solutions,
the
same
and
most
prominent
peak
response
for
perfluorooctanesulfonate
was
used.

No
attempt
was
made
to
quantify
on
the
basis
of
individual
isomeric
components.
The
LOQ
(
limit
of
quantitation)
was
0.200
mg/
L
in
this
study.
The
mean
percent
recovery
of
matrix
fortifications
analyzed
concurrently
during
sample
analysis
was
101.
Samples
collected
at
test
initiation
had
measured
values
from
93.2
to
103%
of
nominal.
Measured
values
for
the
biotic
samples
taken
at
48­
hours
ranged
from
93.6
to
103%
of
nominal,
while
abiotic
samples
ranged
from
105
to
106%
of
nominal.
Measured
values
for
biotic
samples
taken
at
96­
hours
ranged
from
91.4
to
105%
of
nominal,
while
the
abiotic
samples
were
102%
of
nominal.
ENV/
JM/
RD(
2002)
17/
FINAL
204
Summary
of
analytical
chemistry
data:

Nominal
Test
Concentration,
Mg/
L
Measured
Duplicate
Values
at
0,
48,
and
96­
hours,
Respectively,
mg/
L
Mean
Measured
Concentration,
mg/
L
Percent
of
Nominal
Negative
Control
All
<
LOQ
<
LOQ
­
3.1
3.15,
3.02,
2.90,
3.01,2.83,
2.97
3.0
97
6.3
6.22,
6.21,
6.16,
6.43,
6.15,
6.60
6.3
100
13
13.2,
12.1,
12.7,
12.3,
13.1,
12.6
13
100
25
25.0,
25.7,
24.3,
25.7,
25.7,
26.2
25
100
50
49.7,
49.8,
51.1,
51.5,
49.6,
50.8
50
100
50
(
abiotic)
(
1)
53.1,
52.6,
50.9,
51.0
52
104
(
1)
Samples
taken
at
48
and
96­
hours
only
Biological
observations
after
96­
hours:

Fish
in
the
negative
control
and
the
3.0
and
6.3
mg/
L
exposure
concentration
appeared
normal
with
no
mortalities
or
overt
signs
of
toxicity.
All
surviving
fish
in
the
13
and
25
mg/
L
exposures
appeared
normal
with
no
overt
signs
of
toxicity
after
96­
hours.

Cumulative
percent
mortality:

Mean
Measured
Test
Conc.,
mg/
L
24­
hours
48­
hours
72­
hours
96­
hours
Neg.
Control
0
0
0
0
3.0
0
0
0
0
6.3
0
0
0
0
13
0
0
0
20
25
0
0
0
50
50
0
5
35
100
Lowest
concentration
causing
100%
mortality:
50
mg/
L
Mortality
of
controls:
None
CONCLUSIONS
The
potassium
perfluorooctanesulfonate
96­
hour
LC50
for
rainbow
trout
was
determined
to
be
22
mg/
L
with
a
95%
confidence
interval
of
18
 
27
mg/
L.
The
96­
hour
no
mortality
and
no
effects
concentration
was
6.3
mg/
L.

Submitter:
3M,
Environmental
Laboratory,
P.
O.
Box
33331,
St.
Paul,
Minnesota,
55133
ENV/
JM/
RD(
2002)
17/
FINAL
205
DATA
QUALITY
Reliability:
Klimisch
ranking
=
1
REFERENCES
This
study
was
conducted
at
Wildlife
International
Ltd.,
Easton,
MD
at
the
request
of
3M.

OTHER
Last
changed:
1/
24/
02
ENV/
JM/
RD(
2002)
17/
FINAL
206
Robust
Study
Report
Reference
No.
43
­
Perfluorooctanesulfonate,
Potassium
salt
(
PFOS):
96­
Hour
Semi­
Static
Acute
Toxicity
Test
with
the
Sheepshead
Minnow
(





















)
in
saltwater
TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
The
test
substance
is
a
white
powder.
Sample
was
taken
from
3M
lot
number
217.
Sample
was
stored
under
ambient
conditions
prior
to
testing.
Purity
determined
to
be
86.9%
by
LC/
MS,
1
H­
HMR,
19
F­
NMR
and
elemental
analyses
techniques.

METHOD
Method:
OPPTS
850.1075
Type:
Static
renewal
GLP:
Yes
Date
completed:
Study
completed
2001,
report
completed
2002
Species:
Cyprinodon
variegatus
Supplier:
Aquatic
BioSystems,
Inc.,
Fort
Collins,
CO
Analytical
monitoring:
PFOS
measured
at
initiation,
prior
to
and
after
renewal
at
24,
48,
and
72
hours
and
at
test
termination
(
96­
hours)
Exposure
period:
96­
hours
Statistical
methods:
The
use
of
a
single
test
concentration
(
at
water
solubility)
precluded
the
statistical
calculation
of
LC50
values.
Test
fish
age:
Juveniles
Average
Total
Length
and
weight:
3.0
(
2.4
 
3.5)
cm,
0.44
(
0.21­
0.66)
g
Loading:
0.29
g
fish/
L
Pretreatment:
None
Test
conditions:

Dilution
water:
Natural
seawater,
filtered
and
diluted
to
a
salinity
of
approximately
20
parts
per
thousand
with
well
water
Dilution
water
chemistry
(
during
the
4­
week
period
immediately
preceding
the
test):

Salinity:
20
(
20
 
20)
parts
per
thousand
pH:
8.2
(
8.1
 
8.3)

Stock
and
test
solution
preparation:
Primary
stock
prepared
in
methanol
at
40
mg/
L,
sonicated
for
approximately
20
minutes
and
inverted
to
mix
prior
to
use.
After
mixing,
primary
stock
solution
was
proportionally
diluted
with
dilution
water
to
prepare
the
one
test
concentration.
Each
solution
was
stirred
with
a
stainless
steel
whisk
for
approximately
one
minute.
All
test
solutions
appeared
clear
and
colorless.

Solvent:
Methanol
Solvent
concentration
(
treatment
and
solvent
control
groups):
0.5
mL/
L
ENV/
JM/
RD(
2002)
17/
FINAL
207
Concentrations
dosing
rate:
Daily
static
renewal
Stability
of
the
test
chemical
solutions:
Extremely
stable
Exposure
vessels:
25L
polyethylene
aquaria
containing
approximately
15L
of
test
solution;
water
depth
approximately
17.1
cm.

Number
of
replicates:
three
(
biotic),
two
(
abiotic)
Number
of
fish
per
replicate:
ten
Number
of
concentrations:
One
plus
a
negative
and
a
solvent
control,
and
an
abiotic
solution.

Water
chemistry
during
the
study:
Dissolved
oxygen
range
(
0
 
96
hours):
2.8
 
7.4
mg/
L
(
negative
control
exposure)
1.7
 
7.6
mg/
L
(
solvent
control
exposure)
1.6
 
7.6
mg/
L
(
15
mg/
L
exposure)
pH
range
(
0
 
96
hours)
7.9
 
8.3
(
negative
control
exposure)
7.9
 
8.3
(
solvent
control
exposure)
7.9
 
8.3
(
15
mg/
L
exposure)
Test
temperature
range
(
0
 
96
hours)
21.9
 
22.6
º
C
(
negative
control
exposure)
22.1
 
22.9
º
C
(
solvent
control
exposure)
22.2
 
23.1
º
C
(
15
mg/
L
exposure)
­
Method
of
calculating
mean
measured
concentrations:
arithmetic
mean
RESULTS
Nominal
concentrations:
Bk
control,
solvent
control,
20
mg/
L
(
biotic),
20
mg/
L
(
abiotic)

Measured
concentrations:
<
LOQ,
<
LOQ,
15,
13
mg/
L
Element
value:
24­
hour
LC50
=
>
15
mg/
L
(
C.
I.
not
calculable)
48­
hour
LC50
=
>
15
mg/
L
(
C.
I.
not
calculable)
72­
hour
LC50
=
>
15
mg/
L
(
C.
I.
not
calculable)
96­
hour
LC50
=
>
15
mg/
L
(
C.
I.
not
calculable)
All
element
values
based
on
mean
measured
concentrations
Statistical
Evaluation
of
Mortality:
Element
values
and
confidence
interval
could
not
be
calculated
due
to
lack
of
mortality.

Analytical
Methodology:
Analyses
of
test
solutions
were
performed
at
Wildlife
International
Ltd.
using
high
performance
liquid
chromatography
with
mass
spectrometric
detection
(
HPLC/
MS).
When
determining
the
concentration
of
the
test
substance
in
the
test
solutions,
the
same
and
most
prominent
peak
response
for
perfluorooctanesulfonate
was
used.
No
attempt
was
made
to
quantify
on
the
basis
of
individual
isomeric
components.
The
LOQ
(
limit
of
quantitation)
was
5.00
mg/
L
in
this
study.
The
mean
percent
recovery
of
matrix
fortifications
analyzed
concurrently
during
sample
analysis
was
99.2.
Samples
collected
in
exposure
vessels
at
test
initiation
had
measured
values
from
78.6
to
82.0%
of
nominal.
New
samples
collected
at
Hours
24,
48
and
72
had
measured
concentrations
from
74.9
to
81.2
and
86.9
to
92.9,
and
82.9
to
87.1%
of
ENV/
JM/
RD(
2002)
17/
FINAL
208
nominal,
respectively.
Old
samples
collected
at
Hours
24,
48,
and
72
had
measured
concentrations
ranging
from
66.6
to
76.2,
56.4
to
66.9,
and
75.8
to
89.9%
of
nominal,
respectively.
Mean
measured
concentrations
of
PFOS
in
samples
collected
at
test
termination
were
55.6
to
68.4%
of
nominal.
The
measured
concentrations
of
PFOS
from
the
abiotic
treatment
group
were
slightly
lower
than
those
from
the
exposure
treatment
group.
This
may
have
been
due
to
increased
deposition
of
test
substance
at
the
limit
of
solubility,
which
could
have
resulted
from
the
absence
of
the
natural
mixing
action
that
was
provided
by
the
movement
of
the
fish
in
the
exposure
treatment
group.

Summary
of
analytical
chemistry
data:

Nominal
Test
Concentration,
mg/
L
Measured
Replicate
Values
of
old
and
new
Test
Solutions
Respectively,
mg/
L
(
1)
Mean
Measured
Concentration,
mg/
L
Percent
of
Nominal
Negative
Control
All
<
LOQ
<
LOQ
­

Solvent
Control
All
<
LOQ
<
LOQ
­

20
(
2)
16.4,
15.7,
16.0;
15.2,
13.3,
15.2;
16.2,
15.7,
15.0;
13.4,
11.3,
12.7;
17.4,
18.2,
18.6;
15.2,
18.0,
16.2;
17.0,
16.6,
17.4;
13.7,
11.1,
13.4
15
75
20
(
3)

(
abiotic)
10.9,
9.22;
16.1,
16.6;
9.62,
9.69;
16.6,
17.5;
9.01,
9.25;
17.7,
15.9;
9.84,
9.22
13
65
(
1)
Replicate
samples
are
listed
in
this
order:
Day
0
(
new),
24­
hours
(
old),
24­
hours
(
new),
48­
hours
(
old),
48­
hours
(
new),
72­
hours
(
old),
72­
hours
(
new),
96­
hours
(
old)
(
2)
Triplicate
samples
(
3)
Day
0
not
measured,
Duplicate
samples
Biological
observations
after
96­
hours:
Fish
in
the
negative
control
and
solvent
control
appeared
healthy
and
normal
throughout
the
exposure
period.
No
mortalities
appeared
in
the
15
mg/
L
treatment
group
during
the
study.
However,
upon
transfer
to
the
new
test
solution
at
approximately
48
and
72
hours,
some
fish
were
observed
swimming
erratically
and
turning
a
dark
color.
The
fish
appeared
normal
within
approximately
two
hours
after
transfer,
although
one
fish
still
appeared
to
be
discolored
at
test
termination.

Cumulative
percent
mortality:

Mean
Measured
Test
Conc.,
mg/
L
24­
hours
48­
hours
72­
hours
96­
hours
Negative
Control
0
0
0
0
Solvent
control
0
0
0
0
15
0
0
0
0
Lowest
concentration
causing
100%
mortality:
none
 
mortality
limit
apparently
greater
than
solubility
limit.
Mortality
of
controls:
None
ENV/
JM/
RD(
2002)
17/
FINAL
209
CONCLUSIONS
The
potassium
perfluorooctanesulfonate
96­
hour
LC50
for
sheepshead
minnow
was
determined
to
be
>
15
mg/
L,
the
limit
of
solubility
in
this
study.
The
96­
hour
no
mortality
concentration
was
15
mg/
L
and
no
observed
effects
concentration
was
<
15
mg/
L
mg/
L
(
1
fish
out
of
30
was
discolored
at
96­
hours).

Submitter:
3M,
Environmental
Laboratory,
P.
O.
Box
33331,
St.
Paul,
Minnesota,
55133
DATA
QUALITY
Reliability:
Klimisch
ranking
=
1
REFERENCES
This
study
was
conducted
at
Wildlife
International
Ltd.,
Easton,
MD
at
the
request
of
3M.

OTHER
Last
changed:
1/
24/
02
ENV/
JM/
RD(
2002)
17/
FINAL
210
Robust
Study
Report
Number
44.
Bioconcentration
test
of
Salt
(
Na,
K,
Li)
of
perfluoroalkyl
(
C=
4­
12)
sulfonic
acid
[
This
test
was
performed
using
Perfluorooctane
sulfonic
acid,
potassium
salt
(
Test
substance
number
K­
1520)]
in
carp
TEST
SUBSTANCE
Identity:
K­
1520.
Perfluorooctane
sulfonic
acid,
potassium
salt.
Lot
number
A37626B
Supplier:
Kishida
Chemical
Co.,
Ltd.
Remarks:
Test
substance
number
K­
1520.
The
test
substance
is
a
white
powder.
Purity
determined
to
be
100%

METHOD
Method/
guideline
followed:
Method
for
Testing
the
Degree
of
Accumulation
of
Chemical
Substances
in
Fish
Body"
stipulated
in
the
"
Test
Method
for
New
Chemical
Substance"
July
13,
1974,
Revised
October
8,
1998,
No.
5,
Planning
and
Coordination
Bureau,
Environmental
Agency;
No.
615,
Pharmaceutical
Affairs
Bureau,
Ministry
of
Health
and
Welfare:
and
No.
392,
Basic
Industries
Bureau,
Ministry
of
International
Trade
and
Industry,
Japan),
and
Bioconcentration
:
Flow­
through
Fish
test
(
Guideline
305,
June
14,
1996)"
in
the
OECD
Guidelines
for
Testing
of
Chemicals.
Type:
Flow­
through
system
GLP
(
Y/
N):
Yes
Year:
October
13,
2000
 
February
16,
2001
Species:
Carp
(
Cyprinus
carpio)
Supplier:
Fukuokaken
yabegawa
fisherman's
cooperative
association
(
Address:
193­
1
Yamauchi,
Yameshi
Fukuoka
834­
0012,
Japan)
Length
and
weight
at
test
termination:
Mean
length
=
6.4
 
9.6
Mean
weight
=
No
weight
recorded
Loading:
2
and
20
ug/
L
respectively
Fish
Age:
Yearling
fish
Analytical
monitoring:
High­
performance
liquid
chromatography­
Mass
spectrometry
Pretreatment:
The
fish
were
checked
visually
in
the
receiving
and
those
demonstrating
any
abnormality
were
removed.
The
fish
were
reared
for
8
days
in
a
flow
thorough
system
following
an
external
disinfection.
After
rearing,
the
fish
were
medicated
to
eliminate
parasites
and
transferred
to
an
acclimatizing
aquarium.
After
the
second
external
disinfection,
they
were
acclimatized.
The
fish
demonstrating
any
abnormality
during
this
period
were
removed
and
the
remainder
of
the
fish
were
reared
for
15
days
in
a
flow
through
system
at
temperatures
of
25
±
2o
C.
The
fish
were
transferred
to
test
tanks
and
reared
at
the
same
temperature
in
the
flow
through
system
for
another
27
days.
The
fish
were
starved
for
24
hours
before
sampling.
Number
of
concentrations:
Two
plus
a
negative
control
Test
concentration:
(
mean
measured):
Negative
control,
2
and
20
ug/
L
Uptake
period:
58
days
Depuration
period:
37
days
Test
conditions:
Dilution
water:
Groundwater
from
the
premises
of
Kurume
Laboratory
Dilution
water
chemistry:
Specific
conductance:
Not
recorded
Hardness:
111
mg/
L
Alkalinity:
96.1
pH:
7.6
to
7.8
Dissolved
Oxygen:
7.9
to
8.1
ENV/
JM/
RD(
2002)
17/
FINAL
211
Temperature:
25.0
to
25.8
C
Stock
and
test
solution
preparation:
Based
on
preliminary
test
results
for
the
96
hour
LC50
value
and
analytical
detection
limits,
test
concentrations
of
the
test
substance
were
decided
as
follows.
The
control
was
set
as
a
blank
test.
Level
1
was
20
ug/
L
and
Level
2
was
2
ug/
L.
The
test
substance
was
dissolved
with
ion­
exchanged
water
to
prepare
16
and
1.6
mg/
L
stock
solutions.
Diluter
flow
rate:
2
mL/
min
for
stock
solution
and
1600
mL/
min
for
dilution
water;
2307
liters/
day
for
test
water
were
supplied.
Exposure
Vessels:
100
liter
tank
Number
of
replicates:
None
Number
of
fish
per
vessel:
40
Diet:
Nippon
Formula
Feed
Mfg.
Co.,
Ltd.

Water
chemistry
ranges
during
the
study:
Neg.
Control
2
ug/
L
20
ug/
L
Dissolved
Oxygen:
8.0
 
8.1
mg/
L
8.0
 
8.1
mg/
L
7.9
 
8.1
mg/
L
Temperature
C:
25.1
 
25.4
C
25.5
 
25.8
C
25.0
 
25.4
C
pH:
7.6
 
7.8
7.6
 
7.8
7.6
 
7.8
Photoperiod:
Artificial
light
of
white
fluorescent
lamp
(
14
hrs./
day)
Light
intensity:
Artificial
light
of
white
fluorescent
lamp
Collection
of
tissue
samples:
Analysis
of
test
fish
was
performed
six
times
at
each
level
in
duration
of
exposure.
Four
fish
were
taken
out
at
each
sampling
time
and
divided
into
two
groups,
and
then
both
were
analyzed
individually.
Analysis
of
control
fish
was
performed
before
the
experimental
starting
and
after
the
experimental
completion.
Six
fish
were
taken
out
at
each
sampling
time
and
divided
into
three
groups,
and
then
both
were
analyzed
individually.
Because
the
stored
sample
taken
out
from
one
fish
was
too
small
for
the
measurement
of
lipid
content,
a
group
of
two
fish
was
employed.
The
fish
were
separated
into
parts;
tegument,
head
viscera
except
liver,
liver
and
remaining
matter
were
weighted
separately.
The
tegument
consisted
of
the
skin
except
head,
scales,
fin,
alimentary
canal
or
gills.
The
viscera
consisted
of
internal
organs
except
alimentary
canal.
Statistical
methods:
Steady­
state
bioconcentation
BCF
values
calculated
from
the
tissue
concentrations
at
apparent
steady­
state
divided
by
the
mean
water
concentration.
Tissue
concentrations
were
considered
to
be
at
apparent
steady­
state
if
3
or
more
consecutive
sets
of
tissue
concentrations
were
not
significantly
different.

RESULTS
Nominal
concentrations:
Negative
control,
Mean
measured
concentrations:
2
and
20
ug/
L
Bioconcentration
factors
(
BCF):

PFOS
Concentration
in
test
water:
Conc.
After
1
Day
After
7
Days
After14
Days
After
21
Days
After
28
Days
After
43
Days
After
58
Days
Average
(
STD)
20
ug/
L
15.1
14.5
15.7
16.5
17.7
15.4
16.8
16
+
1.12
2
ug/
L
1.78
1.76
1.87
1.93
1.89
1.92
2.01
1.88
+
0.087
PFOS
BCFs
of
Carp
Exposed
to
2
and
20
ug/
L:
Conc.
After
7
Days
After14
Days
After
21
Days
After
28
Days
After
43
Days
After
58
Days
20
ug/
L
260
440
300
690
750
720
ENV/
JM/
RD(
2002)
17/
FINAL
212
2
ug/
L
240
520
410
860
890
1300
Analysis
in
parts
of
test
fish
Level
1
(
20
ug/
L)
BCF
Parts
Tegument
Head
Viscera
Liver
Remainder
Parts
T1
T2
T1
T2
T1
T2
T1
T2
T1
T2
Concentration
(
ng/
g)
22800
1670
0
23400
17500
36000
45300
37800
32000
6260
5380
BCF
1400
1000
1400
1100
2200
2700
2300
1900
380
320
Level
2
(
2
ug/
L)
BCF
Parts
Tegument
Head
Viscera
Liver
Remainder
Parts
T1
T2
T1
T2
T1
T2
T1
T2
T1
T2
Concentration
(
ng/
g)
5490
4750
5600
4730
9900
7410
9190
7650
1810
1390
BCF
2800
2400
2900
2400
5100
3800
4700
3900
930
720
Time
to
reach
50%
clearance:
49
days
at
level
1
and
152
days
at
level
2
Test
organism
mortality:
Negative
control:
None
documented.
Level
1
(
20
ug/
L):
None
documented.
Level
2
(
2
ug/
L):
None
documented.

Analytical
methodology:
Analysis
of
PFOS
in
the
test
water
and
carp
was
performed
using
high­
performance
liquid
chromatography­
mass
spectrometry
(
HPLC­
MS)
analysis.
The
test
water
of
each
level
was
analyzed
once
before
first
analysis
of
test
fish
and
at
the
same
time
as
the
analysis
of
the
test
fish.
Steady
state
was
reached
when
three
successive
analyses
of
BCFs
made
on
samples
taken
at
intervals
of
at
least
48
hours
were
within
+
20%
of
each
other.
When
BCFs
were
less
that
100,
it
was
evaluated
that
a
stead­
state
had
been
reached
after
28
days.

Recoveries
in
water
and
fish
tissues
were
89.3%
and
90.6%
respectively.
Analytical
results
were
corrected
for
procedural
recovery.

CONCLUSIONS
In
this
study,
PFOS
bioconcentrated
in
the
tissues
of
carp.
Test
concentrations
of
2
and
20
ug/
L
were
used.
The
fish
were
exposed
for
58
days
to
makeup
for
excessive
mortality.
Bioconcentration
factors
were
calculated
to
be
720
for
20
ug/
L
and
200
­
1500
for
2
ug/
L.

REFERENCES
Kurume
Laboratory
(
2001).
Chemicals
Evaluation
and
Research
Institute,
Japan.
Test
number:
51520
ENV/
JM/
RD(
2002)
17/
FINAL
213
Annex
3.
Application
of
Equilibrium
Partitioning
Models
to
Determining
Effect
Concentrations
for
PFOS
Salts
in
Soil
and
Sediment
The
review
of
the
complete
set
of
ecotoxicity
test
reports
presented
in
section
2.2.1,
2.3.1
and
2.3.2
has
highlighted
the
absence
of
data
describing
the
toxicity
of
PFOS
salts
to
sediment
and
soil­
dwelling
organisms
and
terrestrial
plants.
Since
tests
with
aquatic
invertebrates
and
plants
have
demonstrated
toxicity
of
PFOS
salts,
and
there
is
generally
a
correlation
between
toxicity
to
aquatic
organisms
living
in
the
water
column
and
toxicity
to
sediment­
and
soil­
dwelling
organisms.
There
is
as
a
consequence
a
need
to
consider
the
possibility
of
deriving
effect
concentrations
for
PFOS
in
soil
and
sediment
to
fill
the
data
gaps
based
on
data
that
already
exist
for
aquatic
organisms.
One
approach
to
deriving
such
data
is
to
use
equilibrium
partitioning
models.

Equilibrium
partitioning
models
are
used
in
two
ways:

 
To
calculate
soil/
sediment
pore
water
concentration
from
overall
soil/
sediment
concentration
 
To
calculate
PNECsoil/
sediment
from
PNECwater.

In
both
cases,
the
equilibrium
constant
for
distribution
between
water
and
soil/
sediment,
Kpsoil/
sediment,
is
needed.
For
many
substances,
Kpsoil/
sediment
is
replaced
by
Koc,
which
is
equivalent
to
assuming
that
only
the
organic
components
of
soil
or
sediment
are
important
in
affecting
the
equilibrium.
However,
for
an
anionic
surfactant,
such
as
PFOS,
it
is
very
likely
that
interaction
with
the
inorganic
substrate
will
also
be
important
(
Salloum
et
al,
2000),
as
has
been
demonstrated
in
a
recent
study
with
PFOS
(
Ellefson).
Therefore
Kp
values
for
an
appropriate
soil
or
sediment
would
have
to
be
used.
It
should
also
be
noted
that
salinity
affects
the
solubility
of
PFOS;
the
high
ionic
content
of
interstitial
water
could
also
affect
the
adsorption
behaviour
of
PFOS.
Furthermore,
it
is
uncertain
over
what
time­
scale
equilibrium
would
be
achieved
(
Ellefson),
and
as
a
consequence
the
extent
of
adsorption
may
be
dependent
upon
the
concentration
of
PFOS
in
the
aqueous
phase.

Whilst
for
all
substances
extrapolation
of
PNEC
from
aquatic
data
to
the
terrestrial
or
sediment
compartment
is
subject
to
uncertainty,
that
uncertainty
is
compounded
when
the
mode
of
uptake
is
different
for
organisms
present
in
different
environmental
compartments.
The
mode
of
toxic
action
and
the
mechanism
of
uptake
of
surfactants
are
complex.
Therefore
the
use
of
equilibrium
partition
models
to
obtain
PNECsoil
or
PNECsediment
is
subject
to
considerable
doubt.

Should
any
laboratory
soil
or
sediment
organism
test
results
become
available
then
there
is
a
further
complication
in
extrapolating
from
these
results
to
PNEC
values
appropriate
to
these
compartments.
The
normal
method
of
extrapolation
is
to
apply
a
correction
that
takes
into
account
the
organic
matter
content
of
the
matrix.
For
substances
that
adsorb
preferentially
to
inorganic
matter
it
would
be
necessary
to
correct
for
both
the
type
of
inorganic
matrix,
the
composition
of
the
aqueous
phase
and
the
concentration
of
the
substance.
This
is
unlikely
to
be
straightforward.

A
search
of
the
open
literature
was
carried
out
in
order
to
assess
the
environmental
fate
and
behaviour
properties
of
substances
with
similar
chemistry
and
properties
to
PFOS.
Published
papers
are
relevant
to
the
consideration
of
equilibrium
partitioning
as
a
relevant
model
for
PFOS
environmental
fate.
Several
papers
were
located
and
the
abstracts
are
summarised
below.
It
is
beyond
the
scope
of
this
work
to
investigate
the
papers
in
detail
but
it
can
be
noted
that
the
technical
quality
of
the
work,
as
with
all
open
literature,
has
already
been
peer
reviewed
(
although
not
subject
to
GLP
audit).
The
findings
of
these
papers
do
not
preclude
the
possibility
of
applying
equilibrium
partitioning
to
PFOS
but
do
highlight
the
technical
difficulties
that
might
be
encountered.
ENV/
JM/
RD(
2002)
17/
FINAL
214
Conclusion
It
is
concluded
that
on
the
basis
of
the
presently
available
data
for
PFOS
that
equilibrium
partitioning
theory
cannot
be
applied
to
determine
either
concentrations
in
interstitial
water
of
soil
and
sediments
or
PNEC
values
for
soil
and
sediments
for
the
following
reasons:

 
the
nature
of
the
adsorption
process
cannot
be
assumed
to
be
linearly
dependent
upon
concentration;
 
the
adsorption
is
likely
to
be
highly
dependent
upon
soil
composition,
particularly
the
inorganic
component;
and
 
the
rate
at
which
equilibrium
might
be
achieved
is
unknown
REFERENCES
Salloum,
M.
J.,
Dudas,
M.
J.,
McGill,
W.
B.
and
Murphy,
S.
M.
(
2000).
Surfactant
sorption
to
soil
and
geologic
samples
with
varying
mineralogical
and
chemical
properties.
Environ.
Toxicol.
Chem.
19(
10),
2436­
2442.

Soil
Adsorption/
Desorption
Study
of
Potassium
Perfluorooctanesulfonate
(
PFOS),
Mark
E.
Ellefson,
3M
Laboratory
Report
No:
E00­
13
11
ENV/
JM/
RD(
2002)
17/
FINAL
215
Review
of
open
literature
Report
title
Summary
of
abstract
Author(
s)
Journal
Terrestrial
risk
assessments
for
linear
alkylbenzenesulfonate
(
LAS)
in
sludgeamended
soils
Reviews
investigations
of
factors
affecting
fate
of
LAS
in
the
terrestrial
environment.
Refers
to
precipitating
effect
of
calcium/
magnesium
ions
in
addition
to
primary
sorption.
Sorbed
form
is
noted
as
being
different
to
the
commercially
available
form
(
for
which
data
were
available).
DeWolfe,
Watze;
Feijtel,
Tom
Chemosphere,
vol
36,
no.
6,
pp
1319­
1343,
1998
Effect
of
sediment
organic­
carbon
on
the
toxicity
of
a
surfactant
to
Hyalella
azteca
Investigation
of
the
effects
of
sediment
organic
carbon
levels
on
the
sorption
and
toxicity
to
Hyalella
azteca
of
anionic
surfactants,
specifically
ABS
(
alkylbenzene
sulfonate).
Studies
indicated
that
higher
levels
of
organic
carbon
led
to
higher
apparent
sorption
coefficients.
A
higher
sediment
concentration
was
required
to
elicit
the
toxic
response
at
higher
organic
carbon
levels.
Cano,
M.
L.;
Dyer,
S.
C.;
Decarvalho,
A.
J.
Environ.
Toxicol.
Chem.,
vol
15,
no.
8,
pp
1411­
1417,
1996
Anionic
surfactant
transport
characteristics
in
unsaturated
soil
Investigations
showed
that
for
an
alkyl
ether
sulfate
and
a
linear
alkylbenzenesulfonate,
soilsurfactant
interactions
were
reversible
and
that
equilibrium
conditions
were
quickly
achieved.
Allred,
Barry;
Brown,
Glenn
O.
Soil
Sci.,
vol
161,
no.
7,
pp
415­
425.
1996
ENV/
JM/
RD(
2002)
17/
FINAL
216
Annex
4.
Summary
of
the
Lowest
Acceptable
Effect
Concentrations
Endpoint
Species
Protocol
Result
(
mg/
l)

Freshwater
fish
­
acute
toxicity
Pimephales
promelas
(
Fathead
minnow)
Not
given
96­
hour
LC50
4.7
Freshwater
fish
­
chronic
toxicity
Pimephales
promelas
(
Fathead
minnow)
OECD
210
&
OPPTS
850.1400
42­
day
NOECsurv/
growth
0.30
Invertebrate
­
acute
toxicity
(
Freshwater)
Daphnia
magna
(
Water
flea)
ASTM
1981
&
OECD
1981
48­
hour
EC50
27
Invertebrate
­
acute
toxicity
(
Salt
water)
Mysidopsis
bahia
(
Mysid
shrimp)
OPPTS
850.1035
96­
hour
LC50
3.6
Invertebrate
­
chronic
toxicity
(
Freshwater)
Daphnia
magna
(
Water
flea)
ASTM
1981
&
OECD
1981
(
Semi­
static)
28­
day
NOECrepro
7
Invertebrate
­
chronic
toxicity
(
Salt
water)
Mysidopsis

bahia
(
Mysid
shrimp)
OPPTS
850.1350
35­
day
NOECrepro/
growth
0.25
Aquatic
plants
 
growth
inhibition
of
freshwater
algae
­
Short­
term
exposure
Selenastrum
capricornutum
(
now







"









	
	












)
OECD
201,
OPPTS
850.5400
&
ASTM
1218­
90E
96­
hour
EC50
cell
density
96­
hour
EbC50
area
under
the
curve
96­
hour
ErC50
growth
rate
96­
hour
NOECgrowth
rate,
cell
density,

area
under
the
growth
curve
72­
hour
EC50
cell
density
72­
hour
EbC50
area
under
the
curve
72­
hour
ErC50
growth
rate
72­
hour
NOECgrowth
rate,
cell
density,

area
under
the
growth
curve
71
71
126
44
70
74
120
70
Aquatic
plants
 
growth
inhibition
of
Saltwater
algae
 
Short­
term
exposure
Skeletonema
costatum
OPPTS
850.5400
96­
hour
EC50
growth
rate
96­
hour
NOECgrowth
rate
>
3.2
>
3.2
Aquatic
plants
 
growth
inhibition
of
freshwater
algae
­
Longer­
term
exposure
Selenastrum
capricornutum
(
now







"









	
	












)
OECD
201,
US
EPA
600/
9­
78­
018
&
ASTM­
E­
35.23
14­
day
EC50
cell
density
14­
day
NOECcell
density
14­
day
EC10
cell
density
95
<
26
16
Freshwater
higher
plants
 
growth
inhibition
Lemna
gibba
OPPTS
850.4400
7­
day
IC50
108
Amphibians
 
embryo
survival,
growth
and
development
Xenopus
laevis
(
African
clawed
frog)
ASTM
E1439­
91
96­
hour
LC50
96­
hour
EC50
malformations
Minimum
concentration
to
inhibit
growth
13.8
12.1
7.97
Sewage
treatment
organisms
Activated
sludge
OECD
209
3­
hour
IC50
respiration
inhibition
>
905
ENV/
JM/
RD(
2002)
17/
FINAL
217
Endpoint
Species
Protocol
Result
(
mg/
kg
of
food)

Dietary
toxicity
to
birds
Anas
platyrhynchos
(
Mallard
duck)
OECD
205,
OPPTS
850.2200
&
FIFRA
E
71­
2
LC50
NOECmortality
NOECbody
weight
628
146
37
Colinus
virginianus
(
Northern
Bobwhite
quail)
OECD
205,
OPPTS
850.2200
&
FIFRA
E
71­
2
LC50
NOECmortality
NOECbody
weight
220
73
73
Endpoint
Species
Protocol
Result
(
µ
g/
bee)

Oral
toxicity
to
bees
Apis
mellifera
(
Honey
bee)
OECD
213,
EPPO
170
72­
hour
LD50
72­
hour
NOEL
0.40
0.21
Contact
toxicity
to
bees
OECD
214,
EPPO
170,
OPPTS
850.3020
(
Contact)
96­
hour
LD50
96­
hour
NOEL
4.78
1.93
ENV/
JM/
RD(
2002)
17/
FINAL
218
Annex
5.
Robust
Summaries
for
Physical
Chemical
Properties
and
Environmental
Fate
Studies
VAPOR
PRESSURE
Title:
Impinger
Studies
of
Volatility
of
FC­
95
and
FC­
143
TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,
1,
2,
2,
3,
3,
4,
4,
5,
5,
6,
6,
7,
7,
8,
8,
8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
The
3M
production
lot
number
was
not
noted.
The
test
sample
is
a
white
powder
of
uncharacterized
purity.

METHOD
Method:
Internal
GLP:
No
Year
study
performed:
1993
Remarks:
Details
outlined
in
the
results
section.
As
described
below,
there
are
a
number
of
methodological
concerns
with
this
study.
In
addition,
the
document
submitted
is
a
combination
of
two
reports.
The
first
report
contained
Parts
I
and
II,
and
a
second
report
contained
Experiment
#
3.
In
addition,
the
first
report
contains
portions
of
two
versions.
As
neither
of
the
first
report
versions
is
dated,
it
is
unknown
which
is
the
final
version.
Part
I
Procedure:
A
mixture
of
10
ppm
FC­
95
(
PFOS)
and
10
ppm
FC­
143
(
PFOA)
was
prepared
in
water
or
water/
isopropanol
solutions
of
the
polar­
organic
compounds
listed
in
the
table
below.
Aliquots
of
each
solution
were
analyzed
in
triplicate
for
fluorochemical
content
by
LC­
thermospray
mass
spectrometry
both
before
and
after
bubbling
280
liters
of
air
through
them.
The
flow
rate
of
the
air
passing
through
the
stock
solutions
was
1
L/
min.
The
solutions
were
kept
in
an
ice
bath
during
bubbling.
The
concentrations
before
and
after
bubbling
were
compared
after
adjusting
for
volume
lost
during
bubbling.
Part
II
Procedure:
Air
was
passed
through
an
apparatus
containing
dry
test
material
and
then
through
glass
wool
at
room
temperature
to
a
chain
of
impingers.
A
50:
50
propanol:
water
solution
containing
500
ppm
ammonium
acetate
was
used
in
the
impingers
to
catch
any
volatilized
PFOS.
All
impingers
were
in
ice
water.
Experiment
#
3
Procedure:
The
same
study
as
in
Part
II
was
conducted
in
duplicate
at
90
°
C.

RESULTS
Vapor
Pressure
Value:
Part
II­
8.7
x
10­
8
torr,
Experiment
#
3­
1.2
x
10­
7
torr
Temperature
°
C:
90
°
C
in
experiment
#
3
Decomposition:
not
stated
ENV/
JM/
RD(
2002)
17/
FINAL
219
Part
I­
Analysis
before
and
after
passing
280
liters
of
air
through
various
stock
solutions.

Results:
Solution
Original
PFOS
Conc.,
ppm
%
PFOS
retained
500
ppm
Tetrabutylammonium
hydroxide
10.0
90
500
ppm
Ammonium
acetate
10.0
71
503
ppm
Laurylpyridinium
chloride
10.0
90
500
ppm
N­
Alkyldimethylbenzylammonium
chloride
10.0
100
500
ppm
Cetyltrimethylammonium
bromide
10.0
95
505
ppm
Tallowtrimethylammonium
chloride
10.0
93
500
ppm
Dicocodimethylammonium
chloride
10.0
84
Water/
1­
Propanol
(
50:
50)
10.0
89
Water/
1­
Propanol
(
50:
50)
0
0
500
ppm
Ammonium
acetate
in
water/
1­
propanol
(
50:
50)
0.10
96
500
ppm
Ammonium
acetate
in
water/
1­
propanol
(
50:
50)
0.20
101
500
ppm
Ammonium
acetate
in
water/
1­
propanol
(
50:
50)
0.40
90
500
ppm
Ammonium
acetate
in
water/
1­
propanol
(
50:
50)
0.80
95
500
ppm
Ammonium
acetate
in
water/
1­
propanol
(
50:
50)
2.0
96
Remarks:
These
findings
appear
to
suggest
a
small
loss
of
PFOS.
However,
in
comments
dated
12/
7/
93,
Dr.
Edwin
Tucker
of
the
Chemistry
Department
at
the
University
of
Oklahoma
indicates
that
it
is
very
unlikely
that
these
flourochemicals
were
removed
by
bubbling
air
through
water
due
to
their
vapor
pressures,
which
are
very
low.
Tucker
thought
more
likely
mechanisms
for
loss
from
the
solution
phase
were
concentration
of
the
surfactants
in
foam
and
loss
from
the
bubbled
solutions
as
foam
or
microdroplets

Part
II­
Analysis
of
impinger
ammonium
acetate
solutions.

Results:
No
test
material
was
found
to
be
present
in
either
the
first
or
second
impinger.
This
indicates
that
any
test
material
transported
from
the
solids
to
air
and
then
into
the
ammonium
acetate
solutions
in
the
impingers
is
below
the
detection
limit.
The
calculated
maximum
pressure
was
8.7
x
10­
8
torr.

Remarks:
In
the
report,
the
maximum
vapor
pressure
calculations
section
contains
errors.
Equation
1
should
use
the
value
0.625:
g,
not
0.625:
g/
mL.
In
addition,
the
"
maximum"
vapor
pressure
calculated
was
erroneously
called
"
minimum"
vapor
pressure
in
the
text
below
both
equations
5
and
7.
Dr.
Edwin
Tucker
of
the
Chemistry
Department
at
the
University
of
Oklahoma
states
that
the
experimental
conditions
do
not
provide
firm
evidence
that
the
number
is
reasonable.
There
is
no
evidence
that
vapor
pressure
equilibrium
was
attained
between
the
solid
and
the
flowing
gas.
ENV/
JM/
RD(
2002)
17/
FINAL
220
Experiment
#
3­
Measuring
Vapor
Pressure
at
90
°
C
Results:
The
report
concludes
that
PFOS
has
a
measurable
vapor
pressure
at
this
temperature.
The
minimum
vapor
pressure
for
the
test
material
was
purported
to
be
1.2
x
10­
7
torr
at
90
°
C.
It
was
considered
a
minimum
because
the
impinger
trains
may
not
have
caught
all
of
the
fluorochemical
that
had
been
volatilized.

Remarks:
There
are
reasons
to
consider
this
a
questionable
result.
No
notation
was
made
about
the
sensitivity
of
the
analytical
measurements
in
this
study,
but
the
quantification
limit
in
the
analysis
in
Part
II
was
0.625
µ
g.
The
concentration
range
from
impinger
train
1
in
Experiment
#
3
was
0.07­
0.075
µ
g.
Impinger
concentrations
ranged
from
0­
0.18
µ
g
in
impinger
train
2.
These
concentrations
are
near
or
below
the
quantification
limit
reported
in
Part
II.

CONCLUSIONS
No
reliable
conclusions
can
be
made
based
on
this
study.
The
general
observation
is
that
this
compound
has
very
low
volatility
or
a
very
low
vapor
pressure
under
ambient
conditions.

Remarks:
none
DATA
QUALITY
Reliabilities:
Klimisch
ranking
3.
There
is
no
information
on
the
validity
of
the
test
method
for
determining
volatility
of
the
test
substance.
This
study
lacks
characterization
of
the
purity
of
the
test
substance.
There
is
no
information
on
the
validity
of
the
analysis
method.

REFERENCE
3M
Environmental
Laboratory.
1993.
3M
Lab
Request
Number
L3306,
3M
Company,
St.
Paul,
MN.
ENV/
JM/
RD(
2002)
17/
FINAL
221
WATER
SOLUBILITY
STUDY
Title:
Solubility
of
PFOS
in
water
TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,
1,
2,
2,
3,
3,
4,
4,
5,
5,
6,
6,
7,
7,
8,
8,
8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
The
test
substance
is
a
white
powder.
The
sample
was
recrystallized
from
a
production
lot
of
FC­
95,
and
assigned
a
test,
control,
and
reference
number
TCR
00017­
046.
The
purity
was
determined
to
be
97.9%
by
LC/
MS,
1H­
NMR,
19F­
NMR
and
elemental
analysis
techniques.

METHOD
Method:
Based
on
OECD
105,
OPPTS
830.7840.

GLP:
Yes
Year
study
performed:
2001
Remarks:
Water
source
is
ASTM
Type
I
water,
Millipore.
The
definitive
test
consisted
of
placing
an
excess
amount
of
test
substance
with
the
appropriate
water
in
centrifuge
tubes.
The
tubes
were
vortexed
and
shaken
at
225
rpm
at
30
°
C
for
24,
48,
or
72
hours
followed
by
24­
hours
of
equilibration
at
24­
25
°
C.
Following
equilibration,
samples
were
centrifuged
and
the
supernatant
was
analyzed
by
high
performance
liquid
chromatography
with
mass
spectrometric
detection
(
LCMS).

RESULTS
Value
(
mg/
L)
at
temperature
°
C:
680
mg/
L
at
24­
25
°
C
Description
of
solubility:
Slightly
soluble
pH
value
and
concentration
at
temperature
°
C:
not
stated
pKa
value
at
25
°
C:
not
stated
Remarks:
The
24­,
48­,
and
72­
hour
solubility
concentrations
were
averaged
to
obtain
the
overall
mean
solubility
concentrations.

CONCLUSIONS
The
overall
mean
solubility
concentration
of
the
test
substance
in
pure
water
was
680
mg/
L.

Remarks:
none
DATA
QUALITY
Reliabilities:
Klimisch
ranking
1
ENV/
JM/
RD(
2002)
17/
FINAL
222
REFERENCE
Ellefson,
M.
2001c.
Solubility
of
PFOS
in
Water.
3M
Company,
3M
Environmental
Laboratory,
Project
Number
E00­
1716.
ENV/
JM/
RD(
2002)
17/
FINAL
223
WATER
SOLUBILITY
STUDY
Title:
Solubility
of
PFOS
in
Natural
Seawater
and
an
Aqueous
Solution
of
3.5%
Sodium
Chloride
TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,
1,
2,
2,
3,
3,
4,
4,
5,
5,
6,
6,
7,
7,
8,
8,
8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
The
test
substance
is
a
white
powder.
The
sample
was
recrystallized
from
a
production
lot
of
FC­
95,
and
assigned
a
test,
control,
and
reference
number
TCR
00017­
046.
The
purity
was
determined
to
be
97.9%
by
LC/
MS,
1H­
NMR,
19F­
NMR
and
elemental
analysis
techniques.

METHOD
Method:
Based
on
OECD
105,
OPPTS
830.7840
GLP:
Yes
Year
study
performed:
2001
Remarks:
The
definitive
test
consisted
of
placing
an
excess
amount
of
test
substance
with
the
appropriate
water
in
centrifuge
tubes.
The
tubes
were
vortexed
and
shaken
at
150
rpm
at
30
°
C
for
24,
48,
and
72
hours
followed
by
24
hours
of
equilibration
at
22­
24
°
C.
Following
equilibration,
samples
were
centrifuged
and
the
supernatant
was
analyzed
by
high
performance
liquid
chromatography
with
mass
spectrometric
detection
(
LCMS).

Water
Sources:
Natural
Seawater
=
Ocean
Scientific,
lot
#
LN
58,
salinity
=
3.5%
Sodium
Chloride
=
EM
Science,
99%
pure,
mixed
with
ASTM
Type
1
water
to
achieve
salinity
of
3.5%

RESULTS
Value
(
mg/
L)
at
temperature
°
C:
Natural
Seawater:
12.4
mg/
L
at
22­
23
°
C;
3.5%
NaCl
Solution:
20.0
mg/
l
at
22­
24
°
C
Description
of
solubility:
Slightly
soluble
pH
value
and
concentration
at
temperature
°
C:
not
stated
pKa
value
at
25
°
C:
not
stated
Remarks:
The
24­,
48­,
and
72­
hour
solubility
concentrations
were
averaged
to
obtain
the
overall
mean
solubility
concentrations
for
the
natural
seawater.
The
24­
hour
values
were
not
included
in
the
mean
solubility
of
the
sodium
chloride
calculation
because
when
the
coefficient
of
variation
was
calculated
for
all
of
the
replicate
analyses
for
that
day,
it
was
>
15%.

CONCLUSIONS
The
overall
mean
solubility
concentration
of
the
test
substance
in
natural
seawater
was
12.4
mg/
L.
In
a
3.5%
NaCl
solution
it
was
20.0
mg/
L.
PFOS
solubility
decreases
with
increasing
ionic
strength
of
the
medium.
ENV/
JM/
RD(
2002)
17/
FINAL
224
Remarks:
none
DATA
QUALITY
Reliabilities:
Klimisch
ranking
1
Remarks:
none
REFERENCE
Ellefson,
M.
2001a.
Solubility
of
PFOS
in
Natural
Seawater
and
an
Aqueous
Solution
of
3.5%
Sodium
Chloride.
3M
Company,
3M
Environmental
Laboratory,
Lab
Project
Number
E00­
1716.
ENV/
JM/
RD(
2002)
17/
FINAL
225
SOLUBILITY
STUDY
Title:
Solubility
of
PFOS
in
Octanol
TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,
1,
2,
2,
3,
3,
4,
4,
5,
5,
6,
6,
7,
7,
8,
8,
8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
The
test
substance
is
a
white
powder.
The
sample
was
recrystallized
from
a
production
lot
of
FC­
95,
and
assigned
a
test,
control,
and
reference
number
TCR
00017­
046.
The
purity
was
determined
to
be
97.9%
by
LC/
MS,
1H­
NMR,
19F­
NMR
and
elemental
analysis
techniques.

METHOD
Method:
Based
on
OECD
105,
OPPTS
830.7840
GLP:
Yes
Year
study
performed:
2001
Remarks:
Octanol
Source:
Aldrich
Screen
Test:
Lot
JU0873804,
99+%
Definitive
Test:
Lot
06238CI,
99.9+%
HPLC
Grade
The
definitive
test
consisted
of
placing
~
0.010
g
test
substance
with
~
10
mL
octanol
in
centrifuge
tubes.
The
tubes
were
shaken
at
~
150
rpm
at
~
30
°
C
for
24,
48,
or
72
hours
followed
by
24
hours
of
equilibration
at
22­
23
°
C.
Following
equilibration,
samples
were
centrifuged
and
the
supernatant
was
analyzed
by
high
performance
liquid
chromatography
with
mass
spectrometric
detection
(
LCMS).

RESULTS
Value
(
mg/
L)
at
temperature
°
C:

24
hours:
56.9
mg/
L
48
hours:
55.7
mg/
L
72
hours:
55.4
mg/
L
Mean
solubility
of
PFOS
in
octanol
=
56.0
mg/
L
Description
of
solubility:
not
stated
pH
value
and
concentration
at
temperature
°
C:
not
stated
pKa
value
at
25
°
C:
not
stated
Remarks:
The
24,
48,
and
72­
hour
solubility
concentrations
were
averaged
to
obtain
the
overall
mean
solubility
concentration.
ENV/
JM/
RD(
2002)
17/
FINAL
226
CONCLUSIONS
The
overall
mean
solubility
concentration
of
the
test
substance
in
pure
octanol
was
56.0
mg/
L.

Submitters'
Remarks:
Typically,
the
Column
Elution
Method
is
recommended
for
use
with
substances
with
solubility
screening
results
of
<
10
mg/
L.
However,
the
shake
flask
method
was
utilized
in
this
study
because
of
the
difficulty
in
obtaining
tubing
compatible
with
octanol
and
the
possible
explosion
hazard
posed
by
possible
leaks
of
a
flammable
solvent
in
an
incubator.

DATA
QUALITY
Reliabilities:
Klimisch
ranking
1
Remarks:
none
REFERENCE
Ellefson,
M.
2001b.
Solubility
of
PFOS
in
Octanol.
3M
Company,
3M
Environmental
Laboratory,
Laboratory
Project
Number
E00­
1716.
ENV/
JM/
RD(
2002)
17/
FINAL
227
SOIL
ADSORPTION
Title:
Pilot
Study
on
Soil
Adsorption
TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
The
sample
was
radiolabeled
(
isotope
and
labeling
position
not
specified).
The
3M
production
lot
number
was
not
noted.
The
test
substance
was
a
white
powder
of
uncharacterized
purity.

METHOD
Method/
guideline
followed:
The
study
explored
the
use
of
autoradiography
of
Thin
Layer
Chromatography
Plates
for
determining
the
soil
mobility
of
PFOS.
This
method
was
developed
by
3M
while
looking
for
alternatives
for
studying
the
adsorption/
desorption
properties
of
the
test
substance.

GLP
(
Y/
N):
No
Year
study
performed:
1978
Statistical
methods:
None
Temperature:
Not
indicated
Remarks:
Little
detail
was
available
regarding
the
study
method.

RESULTS
Results:
The
study
used
TLC
greenhouse
soil
plates.
The
radiolabeled
spot
due
to
PFOS
was
too
faint
to
be
visualized.

CONCLUSIONS
No
conclusion
could
be
reached
in
this
study.

DATA
QUALITY
Reliability:
Klimisch
ranking
3.
These
studies
lacked
sufficient
detail
regarding
methodology.
Isotope
specific
activity
and
chemical/
radiochemical
purity
of
the
test
substance
were
not
provided.
The
method
for
radiosynthesis
was
also
not
provided.
The
analytical
methodology
lacked
validation
and
a
means
of
identifying
and
quantifying
potential
degradation
products.

REFERENCES
Boyd,
S.
A.
1993.
Review
of
Technical
Notebook.
Soil
Thin
Layer
Chromotography.
Number
48277,
p30.
Michigan
State
University.
Mendel,
A.
1978.
Soil
Thin
Layer
Chromatography
 
FC­
95,
FC­
143,
FM­
3422.
Excerpt
from
3M
Technical
Notebook.
October
13,
1978.
Number
48277,
p30.
Project
Number
9970612600.
ENV/
JM/
RD(
2002)
17/
FINAL
228
BIODEGRADATION
STUDY
Title:
Determination
of
Methylene
Blue
Active
Substance
­
FC­
95
TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
The
3M
production
lot
was
158.
The
test
substance
was
a
white
powder
of
uncharacterized
purity.

METHOD
Method/
guideline
followed:
EPA
Method
425.1
Test
Type:
Methylene
Blue
Active
Substance
GLP
(
Y/
N):
No
Year
study
performed:
1989
Contact
time
units:
Not
indicated
Inoculum:
Not
indicated
RESULTS
Degradation
%
after
time:
Not
indicated
Results:

Kinetic
(
for
sample,
positive
and
negative
controls):
Not
indicated
Breakdown
products
(
yes/
no):
Not
indicated
Remarks:
None
CONCLUSIONS
Reviewer's
remarks:
The
test
indicated
that
the
test
sample
exhibited
high
activity
in
this
MBAS
study.

DATA
QUALITY
Reliability:
Klimisch
ranking
3.
This
study
lacked
information
regarding
methodology
and
testing
3M
Lab
Request
Number
MBAS
Result
G1512­
1
835,000
mg/
kg
G1827­
2
831,000
mg/
kg
G1828­
2
730,000
mg/
kg
ENV/
JM/
RD(
2002)
17/
FINAL
229
parameters.
Test
substance
purity
within
the
test
sample
was
not
sufficiently
characterized.
The
sample
description
from
Twin
City
Testing
was
"
liquid",
but
FC­
95
was
a
solid
material.
This
may
indicate
that
the
FC­
95
was
sent
in
a
solution
and
that
the
actual
MBAS
may
be
higher
than
the
values
cited.

REFERENCE
Determination
of
Methylene
Blue
Active
Substance
­
FC­
95.
1989.
Pace
Analytical
and
Twin
City
Testing.
Minneapolis,
Minnesota
 
at
the
request
of
the
3M
Company,
3M
Lab
Request
numbers:
G1512,
G1827,
and
G1828.
ENV/
JM/
RD(
2002)
17/
FINAL
230
SOIL
ADSORPTION
Title:
Soil
Adsorption/
Desorption
Study
of
Potassium
Perfluorooctane
Sulfonate
(
PFOS)

TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
The
test
substance
was
a
white
powder.
The
sample
was
recrystallized
from
a
production
lot
of
FC­
95.
The
purity
was
determined
to
be
97.9%
by
LC/
MS,
1H­
NMR,
19F­
NMR,
and
elemental
analysis
techniques.

METHOD
Method/
guideline
followed:
OECD
106
GLP
(
Y/
N):
Yes
Year
study
performed:
2001
Statistical
methods:
Statistical
analysis
and
plotting
of
the
data
was
done
according
to
OECD
Method
106
using
Microsoft
Excel.

Temperature:
Room
temperature
(
19­
30
°
C)

Stock
and
test
solution
preparation:
The
test
concentrations
and
conditions
were
determined
in
a
preliminary
experiment.
For
the
definitive
experiment,
test
solutions
were
made
by
diluting
a
stock
solution
of
unradiolabeled
perfluorooctanesulfonate
to
a
final
test
substance
concentration
of
approximately
0.5
mg/
L
in
aqueous
0.01
M
CaCl2.
ENV/
JM/
RD(
2002)
17/
FINAL
231
Soil
Characteristics
1Value
is
for
pH
in
water,
not
pH
in
0.1
M
CaCl2
Test
Conditions:

Adsorption
kinetics:
Replicate
study
samples
containing
the
soils
(
or
sediments
or
sludges)
were
equilibrated
by
shaking
for
at
least
12
hours
at
room
temperature
with
0.01M
CaCl2.
Study
samples
were
dosed
with
the
test
substance
at
approximately
0.5
mg/
L
and
placed
on
an
orbital
shaker.
Replicate
sets
of
these
study
samples
were
removed
at
designated
time
points
throughout
a
48­
hour
time
period.
Study
samples
were
then
prepared
and
analyzed
for
the
target
analyte.
The
adsorption
kinetics
were
determined
using
this
data.
The
last
set
of
study
samples
(
48­
hour)
were
saved
and
used
for
the
desorption
kinetics
portion
of
the
method.

Desorption
kinetics
(
one
concentration):
After
the
adsorption
kinetics
experiment,
the
48­
hour
study
samples
were
centrifuged
and
the
aqueous
phase
removed.
The
volume
of
solution
removed
was
replaced
by
an
equal
volume
of
0.01
M
CaCl2
without
test
substance.
The
new
mixture
was
agitated
until
the
desorption
equlibrium
was
reached.
During
a
48­
hour
period,
at
defined
time
intervals,
small
aliquots
of
the
aqueous
phase
were
removed
and
analyzed
for
the
target
analyte.
The
desorption
kinetics
were
determined
using
this
data.
Soil
Class
Clay
Loam
Clay
Loam
Sandy
Loam
River
Sediment
Domestic
Sludge
Source
Agvise
Laboratories,
Northwood,
ND
Agvise
Laboratories,
Northwood,
ND
Agvise
Laboratories,
Northwood,
ND
Agvise
Laboratories,
Northwood,
ND
Agvise
Laboratories,
Northwood,
ND
NIST,
from
Denver,
CO
POTW
Lot
Number
00­
2407
00­
2404
00­
2405
99­
2564
00­
2046
2781
Physical
Description
1.00
mm
airdried
0­
6"
deep
1.00
mm
airdried
0­
6"
deep
1.00
mm
airdried
0­
6"
deep
1.00
mm
airdried
0­
6"
deep
1.00
mm
airdried
0­
6"
deep
200
mesh,
oven­
dried,
sterilized
%
Organic
Carbon
2.6%
4.9%
2.6%
2.8%
1.3%
Not
analyzed
%
Sand
16%
39%
21%
58%
39%
Not
analyzed
%
Silt
22%
50%
46%
22%
42%
Not
analyzed
%
Clay
62%
11%
33%
20%
19%
Not
analyzed
CEC
(
meq/
100g)
54.5
23.9
24.7
23.3
17.5
Not
analyzed
pH
in
0.01
M
CaCl2
7.2
7.4
6.0
7.81
7.7
Not
analyzed
ENV/
JM/
RD(
2002)
17/
FINAL
232
RESULTS
Adsorption
Kinetics
of
PFOS,
1:
5
Soil:
Solution
Ratio,
48­
hour
Time
Point
Soil
Type
Average
Distribution
Coefficient,
Kd,
L/
g
Percentage
of
Organic
Carbon
in
Soil
Average
Organic
Carbon
normalized
Adsorption
Coefficient,
Koc,
L/
g
Clay
0.0183
2.6
70.4
Clay
Loam
0.00972
2.6
37.4
Sandy
Loam
0.0353
2.8
126
River
Sediment
0.00742
1.3
57.1
Domestic
Sludge
<
0.120
Not
available
Not
calculable
All
matrices
adsorbed
the
test
substance
strongly.
The
sludge
demonstrated
very
strong
adsorption
(>
96%)
and
PFOS
was
not
detected
in
the
extracts.
The
data
indicated
that
adsorption
occurred
within
the
first
few
hours
of
exposure
and
the
test
substance
concentration
did
not
vary
significantly
after
16
hours.

Apparent
Desorption
Kinetics
of
PFOS,
1:
5
Soil:
Solution
Ratio,
48­
hour
Time
Point
Soil
Type
Desorption
Coefficient,
Kdes,
L/
g
Barnes
Loam
0.0000471
Clay
Loam
0.0000158
Clay
0.0000349
River
Sediment
0.0000100
Domestic
Sludge
<
0.000237
The
test
substance
was
poorly
desorbed
from
the
soil/
sediment/
sludge
matrices
during
the
48­
hour
study
period.
The
river
sediment
displayed
the
most
desorption
at
39%
after
48
hours.
The
sludge
samples
did
not
desorb
a
detectable
amount
of
test
substance.
Desorption
that
did
occur
was
accomplished
rather
quickly;
after
the
8­
hour
time
point
the
test
substance
concentration
did
not
vary
significantly.

Adsorption
Isotherms
Soil
Type
Log
Kads
F
Kads
F
(
1)
Regression
constant,
1/
n
Regression
Constant,
n
Clay
­
1.2515
0.0560
0.884
1.13
Clay
Loam
­
1.3762
0.0421
0.841
1.19
Sandy
Loam
­
1.0369
0.0919
0.829
1.21
River
Sediment
­
2.0261
0.0094
0.989
1.01
Domestic
Sludge
­
1.246
0.0568
1.2581
0.795
(
1)
Freundlich
adsorption
coefficient
ENV/
JM/
RD(
2002)
17/
FINAL
233
Desorption
Isotherms
Soil
Type
Log
Kdes
F
Kdes
F
(
1)
Regression
Constant,
1/
n
Regression
Constant,
n
Clay
­
0.653
0.222
0.935
1.07
Clay
Loam
­
1.084
0.082
0.954
1.05
Sandy
Loam
­
0.981
0.104
1.01
0.988
River
Sediment
­
1.41
0.039
1.02
0.984
Domestic
Sludge
1.47
29.5
0.327
3.06
(
1)
Freundlich
desorption
coefficient
Freundlich
adsorption
isotherms
were
used
to
relate
the
amount
of
test
substance
adsorbed
on
the
soil
to
the
amount
present
in
the
aqueous
solution
at
equilibrium.
The
values
calculated
for
the
regression
constant
indicate
that
the
data
obtained
for
the
test
substance
over
two
orders
of
magnitude
were
slightly
non­
linear.

CONCLUSIONS
Perfluorooctanesulfonate
(
PFOS)
appeared
to
adsorb
strongly
to
all
of
the
soil/
sediment/
sludge
matrices
tested.
PFOS
would
not
be
considered
to
be
qualitatively
mobile
as
per
OECD
Guideline
106
(
1/
21/
00)
as
the
Kd
values
are
>
1
mL/
g.
The
test
substance,
once
adsorbed,
does
not
desorb
readily,
even
when
extracted
with
an
organic
solvent.
In
either
case,
adsorption
or
desorption,
an
equilibrium
is
achieved
in
less
than
24
hours,
with
substantial
adsorption
(>
50%)
occurring
in
some
of
the
time
0
samples
after
approximately
1
minute
of
contact.
The
test
substance
exhibited
low
mobility
in
all
of
the
adsorbants
tested.
The
shape
of
the
PFOS
adsorption
isotherm
(
H­
type)
indicated
a
very
strong
chemical/
adsorbent
interaction.
Because
PFOS
is
a
strong
acid,
it
likely
forms
strong
bonds
with
soils,
sludge,
and
sediment
via
the
mechanism
of
chemisorption.

DATA
QUALITY
Reliability:
Klimisch
ranking
1.

REFERENCES
Ellefson,
M.
E.
2001d.
Soil
Adsorption/
Desorption
Study
of
Potassium
Perfluorooctanesulfonate
(
PFOS).
3M
Technical
Report.
Project
Number
E00­
1311,
Completion
date
June
4,
2001.
ENV/
JM/
RD(
2002)
17/
FINAL
234
BIOCONCENTRATION
(
PARTITION
COEFFICIENT)

Title:
Kow
(
Solubility
in
Water,
Natural
Seawater,
An
Aqueous
Solution
of
3.5%
Sodium
Chloride,
and
n­
Octanol
with
Subsequent
Calculation
of
the
n­
Octanol
Water
Partition
Coefficient
(
Kow)
of
PFOS
for
each
of
the
Aqueous
Matrices)

TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
The
test
substance
was
a
white
powder.
The
sample
was
recrystallized
from
a
production
lot
of
FC­
95,
and
assigned
the
internal
reference
number
of
TCR­
00017­
046.
Purity
was
determined
to
be
97.9%
by
LC/
MS,
1H­
NMR,
19F­
NMR,
and
elemental
analysis
techniques.

METHOD
Method/
guideline
followed:
Calculated
from
n­
octanol
solubility
and
water
solubility
according
to
OPPTS
830.7550
and
OECD
107.

Test
Type:
n­
Octanol/
Water
partition
coefficient
(
at
saturation)

GLP
(
Y/
N):
Yes
Year
study
performed:
2001
Test
temperature:
n­
Octanol
solubility
value
at
23­
24
°
C
Water
solubility
value
at
24­
25
°
C
Remarks:
The
physical
properties
of
PFOS
did
not
allow
a
determination
of
the
partition
coefficient
by
the
shake
flask
method
per
guidance
provided
in
the
OPPTS
and
OECD
guidelines.
Therefore,
this
study
did
not
bring
the
two
phases
(
n­
octanol
and
water)
into
contact
with
PFOS
at
the
same
time,
and
this
testing
reflected
the
partition
coefficient
for
the
subject
material
at
saturation
only.
The
n­
Octanol/
Water
partition
coefficient
was
calculated
by
dividing
the
solubility
of
PFOS
in
n­
octanol
by
the
solubility
in
water
and
expressing
it
as
the
logarithmic
value.

RESULTS
The
calculated
log
Kow
for
PFOS
was
determined
to
be
 
1.08
at
saturation
(
log(
56
mg/
L
in
n­
octanol/
680
mg/
L
in
water)).

CONCLUSIONS
Remarks:
No
conclusions
could
be
derived
from
this
information.
It
applied
only
to
a
saturated
system,
which
would
not
likely
exist
in
the
environment.

DATA
QUALITY
Reliability:
Klimisch
ranking
1
ENV/
JM/
RD(
2002)
17/
FINAL
235
Remarks:
The
solubility
studies
were
conducted
properly.
However,
the
applicability
of
this
data
point
is
limited.
Application
of
this
value
in
a
risk
assessment
has
limited
or
no
value
as
it
only
applies
to
saturated
systems.

REFERENCES
Kow
(
Solubility
in
Water,
Natural
Seawater,
An
Aqueous
Solution
of
3.5%
Sodium
Chloride,
and
n­
Octanol
with
Subsequent
Calculation
of
the
n­
Octanol
Water
Partition
Coefficient
(
Kow)
of
PFOS
for
each
of
the
Aqueous
Matrices).
2001.
3M
Company.
St.
Paul,
Minnesota.
Environmental
Laboratory
Project
Number
E00­
1716.
ENV/
JM/
RD(
2002)
17/
FINAL
236
BIODEGRADATION
STUDY
Title:
Microbial
Metabolism
(
Biodegradation)
studies
of
Perfluorooctane
Sulfonate
(
PFOS)
II.
Aerobic
Soil
Biodegradation
TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt)
CAS
#
2795­
39­
3
Remarks:
White
powder,
86.9%
purity,
also
called
PFOS
or
FC­
95
METHOD
Method/
guideline
followed
(
experimental/
calculated):
Method
was
designed
by
Springborn
Laboratories,
Inc.

Test
Type
(
aerobic/
anaerobic):
Aerobic
GLP
(
Y/
N):
No
Year
study
performed:
2000
Contact
time/
units:
20
weeks
Inoculum:
The
inoculum
sources
were
soils
collected
from
a
hardwood
forest
in
Hanson,
MA,
a
pine
forest
in
Onset,
MA,
and
a
river
bank
in
Bridgewater,
MA;
and
sediments
collected
from
brackish
sites
below
the
Wareham,
MA
wastewater
treatment
plant
outfall
and
from
the
Narrows
area
in
Wareham,
MA.
Biomass
was
determined
on
day
83
by
both
the
fumigation/
extraction
and
standard
plate
count
methods
and
was
reported
as
17.4
mg
C/
100
g
soil
and
6
x
105
cells/
g,
respectively.

Remarks:
Soil
and
sediment
samples
were
air­
dried,
2.0
mm­
sieved,
and
mixed
together
in
equal
dry
weight
portions.
A
nutrient
mixture
was
prepared
by
combining
a
sterile
potting
soil
extract,
a
trace
mineral
solution,
a
yeast
extract,
and
reagent
water.
The
soil/
sediment
mixture
was
adjusted
to
75%
of
the
water
holding
capacity
using
the
above
nutrient
mixture.
Soil
moisture
was
monitored
weekly
during
the
study,
and
adjusted
using
reagent
water
as
needed.
The
nominal
test
concentration
was
approximately
21.2
mg/
kg.
The
incubation
temperature
was
22
!
3
 
C,
and
the
test
was
conducted
in
the
dark.
Information
on
test
solution
agitation
was
not
provided.
Test
vessels
were
40­
mL
I­
Chem
glass
vials
with
silicone/
Teflonlined
septum
screw
caps
containing
10
g
(
dry
weight)
soil/
sediment
mixture.
Each
test
flask
received
all
components
at
test
initiation.
Samples
were
taken
at
days
7,
14,
21,
28,
35,
42,
49,
56,
and
63.
Entire
samples
were
extracted
with
methanol
via
accelerated
solvent
extraction.
Extracts
passed
through
a
0.2
:
m
nylon
filter
prior
to
analysis.
Samples
were
diluted
as
necessary
in
methanol
and
analyzed
via
LC/
MS.
The
stock
solution
used
to
dose
the
biodegradation
test
systems
was
prepared
at
a
concentration
of
1,060
mg/
L,
which
is
approximately
twice
the
water
solubility
of
PFOS.
No
day
0
samples
were
taken
to
determine
starting
concentrations.

RESULTS
Degradation
%
after
time:
Not
specified
Results:
Essentially
no
PFOS
metabolism
occurred
during
the
study.
ENV/
JM/
RD(
2002)
17/
FINAL
237
Kinetic
(
for
sample,
positive
and
negative
controls):
Not
specified
Breakdown
products
(
yes/
no):
None
indicated
Remarks:
None
CONCLUSIONS
Remarks:
PFOS
is
recalcitrant
in
the
activated
soil/
sediment
system.

DATA
QUALITY
Reliability:
Klimisch
ranking
=
2.
Study
not
conducted
according
to
Good
Laboratory
Practices.
The
stock
solution
used
to
dose
the
systems
was
prepared
at
twice
the
water
solubility
of
PFOS.
No
Day
0
samples
were
taken
to
determine
starting
concentrations.

REFERENCE
The
study
was
conducted
at
Springborn
Laboratories,
Inc.,
Wareham,
Massachusetts,
at
the
request
of
the
3M
Company.
Report
completed
10/
31/
00.
Lab
Project
number
E01­
0434.
ENV/
JM/
RD(
2002)
17/
FINAL
238
BIODEGRADATION
STUDY
Title:
Microbial
Metabolism
(
Biodegradation)
studies
of
Perfluorooctane
Sulfonate
(
PFOS)
III.
Anaerobic
Sludge
Biodegradation
TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt)
CAS
#
2795­
39­
3
Remarks:
White
powder,
86.9%
purity,
also
called
PFOS
or
FC­
95
METHOD
Method/
guideline
followed
(
experimental/
calculated):
Method
was
designed
by
Springborn
Laboratories,
Inc.

Test
Type
(
aerobic/
anaerobic):
Anaerobic
GLP
(
Y/
N):
No
Year
study
performed:
2000
Contact
time/
units:
56
days
Inoculum:
The
inoculum
source
was
an
anaerobic
digestor
at
the
Rockland,
MD
wastewater
treatment
plant.

Remarks:
A
mixture
containing
dried
sludge
extract
(
from
rotating
biological
contacter
wastewater
treatment,
Bridgewater,
MA),
unspecified
OECD
mineral
media,
and
resazurin
indicator
served
as
the
test
medium.
The
media
were
prepared
under
nitrogen
purge
to
exclude
oxygen.
The
nominal
test
concentration
was
approximately
20.8
mg/
L,
and
loading
in
test
vessels
was
300
mL
anaerobic
sludge
per
liter
of
medium.
The
test
temperature
was
35
 
C,
and
the
test
was
conducted
in
the
dark.
Information
on
test
solution
agitation
was
not
provided.
Twenty
160­
mL
serum
bottles
containing
100
mL
of
test
solution
(
purged
with
nitrogen
after
filling)
with
crimped
butyl
rubber
tops
served
as
test
flasks.
Each
flask
received
all
components
at
test
initiation.
Samples
were
taken
from
the
test
bottle
on
days
7,
14,
21,
28,
35,
42,
49,
and
56,
and
from
inoculum
control
bottles
on
days
7
and
56.
For
analysis,
an
aliquot
was
removed
from
each
bottle
and
centrifuged.
Both
the
supernatant
and
the
solid
biomass
portion
were
analyzed
via
LC/
MS.
The
stock
solution
used
to
dose
the
biodegradation
test
systems
was
prepared
at
a
concentration
of
1,060
mg/
L,
approximately
twice
the
water
solubility
of
PFOS.
No
day
0
samples
were
taken
to
determine
starting
concentrations.

RESULTS
Degradation
%
after
time:
Not
specified
Results:
No
apparent
PFOS
biodegradation
occurred
over
the
56­
day
period.
ENV/
JM/
RD(
2002)
17/
FINAL
239
Kinetic
(
for
sample,
positive
and
negative
controls):
Not
specified
Breakdown
products
(
yes/
no):
None
indicated
Remarks:
None
CONCLUSIONS
Remarks:
PFOS
is
recalcitrant
in
the
anaerobic
system.

DATA
QUALITY
Reliability:
Klimisch
ranking
=
2.
Study
not
conducted
according
to
Good
Laboratory
Practices.
The
stock
solution
used
to
dose
the
systems
was
prepared
at
twice
the
water
solubility.
No
Day
0
samples
were
taken
to
determine
starting
concentrations.

REFERENCE
The
study
was
conducted
at
Springborn
Laboratories,
Inc.,
Wareham,
Massachusetts,
at
the
request
of
the
3M
Company.
Report
completed
10/
31/
00.
Lab
Project
number
E01­
0434.
ENV/
JM/
RD(
2002)
17/
FINAL
240
BIODEGRADATION
STUDY
Title:
Microbial
Metabolism
(
Biodegradation)
studies
of
Perfluorooctane
Sulfonate
(
PFOS)
IV.
Pure
Culture
Study
TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt)
CAS
#
2795­
39­
3
Remarks:
White
powder,
86.9%
purity,
also
called
PFOS
or
FC­
95
METHOD
Method/
guideline
followed
(
experimental/
calculated):
Two
study
types
were
used
(
Pure
Culture
and
Closed
Vial
Headspace)
and
Springborn
Laboratories,
Inc./
Betts
et
al.,
1974
is
cited.

Test
Type
(
aerobic/
anaerobic):
Aerobic
GLP
(
Y/
N):
No
Year
study
performed:
2000
Pure
Culture
Studies
Contact
time/
units:
7
days
Inoculum:
Four
separate
pure
cultures
from
the
American
Type
Culture
Collection
(
ATCC)
were
tested:
Cunninghamella
echinulata
var.
echinulata
(
fungi,
ATCC
#
9244)
Mucor
circinelloides
f.
griseocyanus
(
fungi,
ATCC#
1207a)
Phanerochaete
chrysosporium
(
fungi,
ATCC
#
24725)
Streptomyces
griseus
(
actinomycete,
ATCC
#
13273)

Remarks:
The
test
medium
was
soybean
grits­
glucose
(
SGG)
and
test
vessel
loading
was
6
mL
of
Stage
II
cultures
into
60
mL
media,
with
24
hours
of
agitation
on
a
shaker
table
at
250
rpm
prior
to
the
addition
of
PFOS.
The
nominal
test
concentration
was
approximately
20.9
mg/
L.
The
test
temperature
was
26
 
C.
The
test
vessel
type
was
not
noted.
Each
test
flask
received
all
necessary
components
at
test
initiation.
All
work
utilized
strict
aseptic
technique
until
harvest
at
day
7.
Samples
were
taken
on
days
0
and
7.
Analysis
of
the
broth
and
cells
collected
via
centrifugation
was
performed
by
LC/
MS.
The
stock
solution
used
to
dose
the
biodegradation
test
systems
was
prepared
at
a
concentration
of
1,060
mg/
L,
approximately
twice
the
water
solubility
of
PFOS.

Degradation
%
after
time:
Not
specified
Results:
The
studies
with
Cunninghamella,
Mucor,
and
Streptomyces
did
not
provide
any
indication
of
biotransformation
of
PFOS.
Possible
biotransformation
of
PFOS
(
90%
mass
balance)
by
Phanaerochaete
was
noted,
and
closed
vial
studies
were
performed
to
confirm
this.

Kinetic
(
for
sample,
positive
and
negative
controls):
Not
specified
ENV/
JM/
RD(
2002)
17/
FINAL
241
Breakdown
products
(
yes/
no):
None
indicated
Remarks:
None
Closed
Vial
Headspace
Study
Contact
time/
units:
3
days
Inoculum:
Phanerochaete
chrysosporium
(
fungi,
ATCC
#
24725)

Remarks:
The
test
medium
was
soybean
grits­
glucose
(
SGG)
medium
at
1/
10
and
1/
100
strength
plus
resazurin.
The
test
concentration
was
0.2
mg/
L.
The
incubation
temperature
was
26
 
C,
and
a
shaker
table
in
an
environmental
chamber
at
250
rpm
was
used
in
incubation.
Sterile
22­
mL
vials
served
as
test
vessels.
Initial
inoculum
test
vessel
loading
details
were
not
noted.
Each
test
flask
received
all
necessary
components
at
test
initiation.
Headspace
was
purged
with
oxygen
and
vials
were
immediately
crimped.
Samples
were
taken
once
on
day
3.
Analysis
of
the
broth
and
cells
collected
via
centrifugation
was
performed
by
LC/
MS.
The
stock
solution
used
to
dose
the
biodegradation
test
systems
was
prepared
at
a
concentration
of
1,011
mg/
L,
approximately
twice
the
water
solubility
of
PFOS.
No
initial
measured
concentrations
were
taken
in
this
closed
vial
study.
Difficulties
were
encountered
in
maintaining
aerobicity
and
only
3
days
of
exposure
were
maintained.

RESULTS
Degradation
%
after
time:
Not
specified
Results:
The
results
indicated
no
significant
biotransformation
of
PFOS
by
Phanaerochaete
fungi.

Kinetic
(
for
sample,
positive
and
negative
controls):
Not
specified
Breakdown
products
(
yes/
no):
None
indicated
Remarks:
None
CONCLUSIONS
Remarks:
It
did
not
appear
that
the
four
species
were
capable
of
metabolizing
PFOS.

DATA
QUALITY
Reliability:
Klimisch
ranking
=
2.
The
stock
solution
used
to
dose
the
systems
was
prepared
at
twice
the
water
solubility.
There
were
no
initial
measured
concentrations
taken
in
the
closed
vial
study.
These
studies
were
not
conducted
in
accordance
with
Good
Laboratory
Practices.

REFERENCE
The
study
was
conducted
at
Springborn
Laboratories,
Inc.,
Wareham,
Massachusetts,
at
the
request
of
the
3M
Company.
Report
completed
11/
3/
00.
Lab
Project
number
E01­
0434.
ENV/
JM/
RD(
2002)
17/
FINAL
242
BIODEGRADATION
STUDY
Title:
The
18­
day
aerobic
biodegradation
study
of
perfluorooctanesulfonyl­
based
chemistries.

TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
C8F17SO3
­
K+.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS#
2795­
39­
3)

Remarks:
PFOS
is
a
white
powder.
The
original
3M
product
lot
number
was
not
noted.
The
PFOS
was
HPLC
purified
and
assigned
3M
standard
identification
#
TCR­
00017­
046.
Upon
receipt
at
the
testing
laboratory,
the
test
article
was
given
a
test,
control,
and
reference
(
TCR)
number
CA­
TCR02­
014.
The
submitter
suggests
that
an
Interim
Certificate
of
Analysis
reports
the
purity
to
be
97.9%.
All
results
in
the
study
were
calculated
assuming
100%
purity.
However,
in
the
study
report,
the
purity
is
noted
as
86.4%.
No
explanation
of
this
discrepancy
was
given.

METHOD
Method/
guideline
followed:
Based
on
EPA
guidelines
OPPTS
835.3200
Test
Type:
Aerobic
GLP:
No
Year
study
performed:
2001
Contact
time:
18
days
Inoculum:
Activated
sludge
collected
7/
31/
00
from
the
aeration
basin
at
the
Metro
Wastewater
Treatment
Plant,
St.
Paul,
MN.
The
MLSS
was
determined
to
be
2,280
mg/
L
when
first
collected.
The
MLSS
was
stored
at
4
º
C
for
approximately
5
weeks
prior
to
being
used
for
this
study.
The
sludge
was
allowed
to
settle
and
the
solids
used
for
inoculum.
The
settled
sludge
constituted
approximately
20%
of
the
volume
(~
200
mL)
of
the
MLSS
used.

Test
Medium:
Test
flasks
were
prepared
using
a
mineral
salts
medium
defined
in
EPA
Guideline
OPPTS
835.3200.
Methanol
(
1
mL
per
liter)
was
added
per
liter
of
mineral
medium.
Fifty
mL
of
settled
sludge
was
added
per
liter
of
mineral
salts
medium.
Mineral
medium
plus
sludge
was
prepared
9/
7/
00,
while
fresh
medium
without
sludge
(
abiotic
controls)
was
prepared
8/
10/
00.

Remarks:
Three
types
of
samples
were
prepared
for
this
study:
blank
sludge
controls
(
mineral
medium,
inoculum),
abiotic
controls
(
mineral
medium,
PFOS),
and
test
substance
samples
(
mineral
medium,
inoculum,
PFOS).
The
test
vessels,
sterile
125
mL
Nalgene
polycarbonate
culture
flasks
containing
25
mL
of
media,
were
set
in
duplicate.
Additional
quality
control
samples
(
blanks)
were
prepared
and
analyzed
as
appropriate.

The
test
concentration
used
was
2.455
mg/
L.
The
samples
were
agitated
at
~
200
rpm
at
a
temperature
of
25
±
3
º
C.
The
test
vessels
were
spiked
with
6
µ
L
of
a
10,230
mg/
L
solution
of
PFOS
in
methanol
yielding
2.455
mg/
L.
Sampling
was
done
on
days
0
and
18.
The
day
0
test
vessels
were
prepared
and
immediately
placed
in
a
freezer
that
was
maintained
at
 
20
º
C
until
analyzed.
After
18
days,
the
test
vessels
were
removed
from
the
incubator
and
frozen
until
final
sample
preparation
by
solid
phase
extraction
(
SPE).
Following
thawing,
test
vessel
contents
were
adjusted
to
1%
acetic
acid
and
then
passed
through
a
ENV/
JM/
RD(
2002)
17/
FINAL
243
conditioned
SEP­
VAC
C18
6cc
SPE
cartridge.
Methanol
was
then
added
to
the
emptied
culture
flask,
shaken
vigorously
and
then
passed
through
the
SPE
cartridge
to
extract
adsorbed
analytes.
A
second
methanol
wash
was
collected
separately
for
analysis
to
ensure
quantitative
extraction.

Quantitative
analysis
was
conducted
on
an
HP1100
high
performance
liquid
chromatograph
with
mass
spectrometer
detector
(
HPLC/
MSD)
system.
The
MSD
was
operated
in
electrospray
ionization
in
negative­
ion
mode
using
selected­
ion
monitoring
(
SIM)
for
quantitation.

In
addition
to
PFOS,
the
additional
compounds
quantified
are
specified
below.
In
the
case
of
the
compounds
that
are
potassium
or
ammonia
salts,
only
the
concentration
of
the
fluorochemical
anion
was
quantified
and
reported.

Compound
Name
Acronym
Chemical
Formula
2­(
N­
ethyl
Perfluorooctane
sulfonamido)
ethyl
alcohol
N­
EtFOSE
Alcohol
C8F17SO2N(
C2H5)
CH2CH2OH
2­(
N­
ethyl
Perfluorooctane
sulfonamido)
acetic
acid
N­
EtFOSAA
C8F17SO2N(
C2H5)(
CH2COOH)

2­(
Perfluorooctane
sulfonamido)
acetic
acid
M556
C8F17SO2NH(
CH2COOH)

N­
Ethyl
perfluorooctane
sulfonamide
N­
EtFOSA
C8F17SO2NH(
C2H5)
Perfluorooctane
sulfinate,
potassium
salt
PFOSulfinate
C8F17SO2
­
K+

Perfluorooctanoate,
ammonium
salt
PFOA
C8F15COO­
NH4
+

Perfluorooctane
sulfonamide
FOSA
C8F17SO2NH2
No
reference
substance
was
used.
However,
when
the
results
from
an
EtFOSE
alcohol
study
conducted
at
the
same
time
are
compared
to
the
previous
EtFOSE
alcohol
35­
day
study,
the
viability
of
the
microbial
inoculum
is
confirmed.

RESULTS
Degradation
%
after
time:
0
Results:
After
18
days,
the
analytical
results
demonstrate
that
after
exposure
to
municipal
wastewater
treatment
sludge,
2.455
mg/
L
PFOS
was
not
measurably
degraded
biotically
or
abiotically.
Mass
balance
for
PFOS
test
vessels
was
excellent
and
ranged
from
104­
108%.

Breakdown
products:
No
Remarks:
None
CONCLUSIONS
No
loss
of
PFOS
was
demonstrated.
Mass
balance
was
104­
108%.
The
results
from
this
study
confirm
the
results
from
other
aerobic
biodegradation
studies
of
PFOS.
Submitters'
Remarks:
The
submitters
suggested
a
Klimisch
data
quality
ranking
of
2
because
the
study
was
conducted
as
a
non­
GLP
study,
but
with
the
understanding
that
good
data
quality
objectives
be
met.
ENV/
JM/
RD(
2002)
17/
FINAL
244
REFERENCE
Lange,
C.
2001a.
The
18­
day
aerobic
biodegradation
study
of
perfluorooctanesulfonyl­
based
chemistries.
Pace
Analytical
Services,
Inc.
Minneapolis,
MN.
3M
Company.
Minneapolis,
MN.
ENV/
JM/
RD(
2002)
17/
FINAL
245
STABILITY
IN
WATER
STUDY
Title:
Hydrolysis
Reactions
of
Perfluorooctane
Sulfonate
(
PFOS)

TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate­
potassium
salt.
May
also
be
referred
to
as:
PFOS,
PFOS­
potassium
salt,
1­
perfluorooctanesulfonic
acid­
potassium
salt,
or
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluorosulfonate­
potassium
salt
(
CAS
#
2795­
39­
3).

Remarks:
3M
production
lot
number
was
171.
Test
substance
is
a
light­
colored
powder
at
25
º
C.

METHOD
Method/
guideline
followed:
Based
on
OPPTS:
835.2110.

Test
Type:
Hydrolysis
as
a
function
of
pH
GLP
(
Y/
N):
No
Year
study
performed:
2001
Test
sample
preparation:
Test
solutions
consisted
of
1.0
mL
buffered
aqueous
solutions
at
6
pH
levels
(
1.5,
3.0,
5.0,
7.0,
9.0,
11.0).
The
resulting
PFOS
concentration
in
all
test
samples
(
sample
triplicates
and
matrix
spike
samples)
was
approximately
500
µ
g/
L.
Samples
were
shielded
from
light
during
incubation
at
50
º
C
for
periods
of
0
to
49
days.
Control
samples
and
blanks
addressed
potential
non­
hydrolytic
degradation
routes.

Analytical
Procedures:
Samples
were
analyzed
by
quantitative
HPLC/
MS.

Remarks:
This
study
was
conducted
at
50
º
C
in
order
to
facilitate
hydrolysis.
Rates
derived
at
50
º
C
were
extrapolated
to
25
º
C
by
dividing
by
a
factor
of
10,
which
is
valid
for
reactions
with
Arrhenius
heats
of
activation
near
18
kcal/
mole.

RESULTS
Degradation
%:
Not
applicable;
no
degradation
was
reported.

Half­
life
(
t1/
2):









	
Breakdown
products
(
yes/
no):
No
Remarks:
The
analytical
results
indicate
no
degradation
of
PFOS
or
dependence
on
pH.
The
mean
and
standard
deviation
of
all
observed
PFOS
concentrations,
pooled
over
the
6
observed
pH
levels,
indicate
that
the
pseudo­
first
order
hydrolytic
half­
life
of
PFOS
is
greater
than
41
years.

CONCLUSIONS
The
analytical
results
indicate
no
degradation
of
PFOS
or
dependence
on
pH.
The
study
indicates
that
the
hydrolytic
half­
life
of
PFOS
in
water
is
greater
than
41
years.
ENV/
JM/
RD(
2002)
17/
FINAL
246
Submitters'
Remarks:
The
authors
assigned
a
Klimisch
ranking
of
2
for
the
reliability
of
this
study.
The
authors
also
noted
that
the
study
was
well­
conducted,
but
not
under
GLP.

Reviewer's
Remarks:
None
REFERENCE
Hatfield,
T.
2001a.
Hydrolysis
Reactions
of
Perfluorooctane
Sulfonate
(
PFOS).
3M
Environmental
Laboratory
and
Pace
Analytical
Services.
Lab
request
number
W1878.
Minneapolis,
MN.
ENV/
JM/
RD(
2002)
17/
FINAL
247
BIODEGRADATION
STUDY
Title:
The
35­
day
aerobic
biodegradation
study
of
PFOS
TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
C8F17SO3
­
K+.
(
1­
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS#
2795­
39­
3)

Remarks:
PFOS
is
a
white
powder.
The
original
3M
production
lot
number
was
not
noted.
The
PFOS
was
HPLC
purified.
An
Interim
Certificate
of
Analysis
reports
the
purity
to
be
97.9%.
All
results
in
this
report
are
based
on
a
purity
of
86.4%,
however.
The
lower
purity
value
was
associated
with
a
standard
the
laboratory
was
using
during
this
study.

METHOD
Method/
guideline
followed:
Based
on
EPA
Guidelines
OPPTS
835.3200,
OPPTS
835.3210,
OPPTS
835.5045.

Test
Type:
Aerobic
GLP:
No
Year
study
performed:
2001
Contact
time:
35
days
Inoculum:
Activated
sludge
collected
9/
18/
00
from
the
aeration
basin
at
the
Metro
Wastewater
Treatment
Plant,
St.
Paul,
MN.
The
suspended
solids
were
allowed
to
settle
for
approximately
2
days
at
4 C
and
the
settled
sludge,
approximately
20%
of
the
volume,
was
used
to
prepare
cultures
for
the
biodegradation
study.
The
mixed
liquor
suspended
solids,
MLSS,
was
not
noted.

Test
medium:
Test
flasks
were
prepared
using
a
mineral
salts
medium
defined
in
EPA
Guideline
OPPTS
835.3200.
Fifty
mL
of
settled
sludge
was
added
per
liter
of
mineral
salts
medium.

Remarks:
Three
types
of
samples
were
prepared
for
this
test:
blank
sludge
controls
(
mineral
medium,
inoculum),
abiotic
controls
(
mineral
medium,
PFOS),
and
test
substance
samples
(
mineral
medium,
inoculum,
PFOS).

Test
vessels,
sterile
125
mL
Nalgene
polycarbonate
culture
flasks
containing
25
mL
media,
were
set
in
duplicate.
Additional
quality
control
samples
(
blanks)
were
prepared
and
analyzed
as
appropriate.

The
test
concentration
used
was
2.582
mg/
L.
The
samples
were
agitated
at
200
rpm
at
25
±
3
º
C.
Test
vessels
were
spiked
6
µ
L
of
10,760
mg/
L
solution
of
PFOS
in
methanol
yielding
a
2.582
mg/
L
PFOS
solution.
Samples
were
taken
on
days
0,
2,
5,
7,
14,
and
35.

The
day
zero
test
vessels
were
prepared
and
immediately
placed
in
a
freezer
that
was
maintained
at
 
20
±
7
º
C.
Upon
removal
from
the
incubator,
test
vessels
on
other
days
were
either
immediately
frozen,
or
prepared
by
solid
phase
extraction
(
SPE).
Following
thawing,
if
needed,
test
vessel
contents
were
adjusted
to
1%
acetic
acid
and
then
passed
through
a
conditioned
SEP­
VAC
C18
6cc
SPE
cartridge
containing
a
plug
of
quartz
wool
to
deter
plugging.
Methanol
was
then
added
to
the
emptied
culture
flask,
shaken
ENV/
JM/
RD(
2002)
17/
FINAL
248
vigorously
and
then
passed
through
the
SPE
cartridge
to
extract
adsorbed
analytes.
A
second
methanol
wash
was
then
collected
separately
for
analysis
to
ensure
quantitative
extraction.

Quantitative
analysis
was
conducted
on
an
HP1100
high
performance
liquid
chromatograph
with
mass
spectrometer
detector
(
HPLC/
MSD)
system.
The
MSD
was
operated
in
electrospray
ionization
in
negative­
ion
mode
using
selected­
ion
monitoring
(
SIM)
for
quantitation.

In
addition
to
the
parent,
PFOS,
the
compounds
below
were
quantified.
In
the
case
of
the
compounds
that
are
potassium
or
ammonia
salts,
only
the
concentration
of
the
fluorochemical
anion
was
quantified
and
reported.

Compound
Name
Acronym
Chemical
Formula
2­(
N­
ethyl
Perfluorooctane
sulfonamido)
ethyl
alcohol
N­
EtFOSE
Alcohol
C8F17SO2N(
C2H5)
CH2CH2OH
2­(
N­
ethyl
Perfluorooctane
sulfonamido)
acetic
acid
N­
EtFOSAA
C8F17SO2N(
C2H5)(
CH2COOH)

2­(
Perfluorooctane
sulfonamido)
acetic
acid
M556
C8F17SO2NH(
CH2COOH)

N­
Ethyl
perfluorooctane
sulfonamide
N­
EtFOSA
C8F17SO2NH(
C2H5)
Perfluorooctane
sulfinate,
potassium
salt
PFOSulfinate
C8F17SO2
­
K+

Perfluorooctanoate,
ammonium
salt
PFOA
C8F15COO­
NH4
+

Perfluorooctane
sulfonamide
FOSA
C8F17SO2NH2
RESULTS
Degradation
%
after
time:
0
Results:
The
analytical
results
demonstrate
that
when
exposed
to
municipal
wastewater
treatment
sludge
for
35
days,
the
2.582
mg/
L
PFOS
samples
generated
no
quantifiable
degradation
products.
PFOS
was
recovered
at
2.553
±
0.102
mg/
L
in
the
pooled
study
samples
and
at
2.653
±
0.083
mg/
L
in
the
pooled
abiotic
samples.
The
measured
concentration
of
PFOS
was
always
100
±
7%
of
the
expected
concentration.

Breakdown
products:
No
Remarks:
None
CONCLUSIONS
The
six­
sample
point
screening
study
established
that
PFOS
is
not
biodegraded
by
the
microbial
populations
of
the
municipal
waste
treatment
inoculum
used
under
the
conditions
tested.

Submitters'
remarks:
The
submitters'
assigned
a
data
quality
rating
of
2
for
this
study
since
it
was
conducted
as
a
non­
GLP
study,
but
with
the
understanding
that
good
data
quality
objectives
be
met.

No
sample
matrix
spikes
were
included
in
this
study.
However,
PFOS
was
recovered
at
expected
concentrations,
and
previous
duplicated
results
obtained
during
a
related
project
showed
excellent
PFOS
recoveries
from
sludge
using
the
same
extraction
method.
ENV/
JM/
RD(
2002)
17/
FINAL
249
A
series
of
positive
controls
were
not
run
with
this
study.
However,
the
sludge
used
in
this
study
was
also
used
for
the
preparation
of
samples
in
other
biodegradation
studies,
and
in
some
of
those
studies,
preliminary
results
demonstrated
positive
biological
activity
for
degradation
of
test
samples.

REFERENCE
Lange,
C.
2001b.
The
35­
day
aerobic
biodegradation
study
of
PFOS.
Pace
Analytical
Services,
Inc.
Minneapolis,
MN.
3M
Company.
Minneapolis,
MN.
ENV/
JM/
RD(
2002)
17/
FINAL
250
PHOTODEGRADATION
STUDY
Title:
Screening
studies
on
the
aqueous
photolytic
degradation
of
potassium
perfluorooctane
sulfonate
(
PFOS)

TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate­
potassium
salt.
May
also
be
referred
to
as:
PFOS,
PFOS­
potassium
salt,
1­
perfluorooctanesulfonic­
potassium
salt,
or
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoropotassium
salt.

Remarks:
3M
production
lot
number
171.
The
test
substance
is
a
light­
colored
powder
at
25
º
C.
The
purity
was
determined
to
be
86.4%
by
LC/
MS,
1H­
HMR,
19F­
NMR,
and
elemental
techniques.

METHOD
Method/
guideline
followed:
Based
on
OPPTS:
835.5270
and
OECD
Draft
Document
"
Phototransformation
of
Chemicals
in
Water
 
Direct
and
Indirect
Photolysis,"
August
2000.

Test
Type:
Direct
and
indirect
photolysis
GLP
(
Y/
N):
No
Year
study
performed:
2001
Light
Source:
Suntest
CPS+
or
Suntest
XLS+
lamp
Light
Spectrum
(
nm):
290­
800
nm
Relative
Intensity
based
on
Intensity
of
Sunlight:
680
w/
m2
Spectrum
of
substance
(
max
lambda,
max
epsilon
and
epsilon
295):
A
UV/
Vis
spectrum
of
a
saturated
aqueous
solution
of
PFOS
was
recorded
between
190
and
110
nm.

Test
sample
preparation:
Aliquots
of
PFOS
were
added
to
3
separate
sets
of
VOA
screw
cap
vials
(
exposed,
unexposed,
and
control
vials)
containing
5
mL
of
appropriate
matrix.
Test
vials
were
placed
in
the
photoreactor.
Control
vials
were
wrapped
in
aluminum
foil,
sealed
in
a
plastic
bag,
and
placed
in
the
photoreactor.

Analytical
Procedures:
Samples
were
analyzed
by
quantitative
LC/
MS
and
GC/
MS
techniques.

Remarks:
The
duration
of
the
study
was
67­
167
hours.
The
test
media
used
were
water,
H2O2/
water
(
1:
1
molar
equivalent),
Fe2O3/
water
(
Fe3+
at
24X
molar
excess),
Fe2O3/
water
with
H2O2,
and
commercial
(
Aldrich)
humic
material
prepared
as
in
OPPTS
835.5270.
All
tests
included
a
series
of
unexposed
controls
(
kept
in
the
dark)
for
the
evaluation
of
any
degradation
reactions
occurring
without
the
presence
of
light.
Solvent,
matrix,
and
control
blanks
and
spikes
were
tested
under
each
condition.

RESULTS
Concentration
of
Substance:
Not
stated
ENV/
JM/
RD(
2002)
17/
FINAL
251
Temperature
 C:
25
±
3
Direct
photolysis:
No
decomposition
was
observed.

Indirect
photolysis:
Data
obtained
from
the
Fe2O3
matrix
samples
(
with
and
without
H2O2)
were
pooled
to
provide
sufficient
data
to
estimate
the
minimum
half­
life.
The
mean
standard
deviation
of
these
data
indicate
that
the
minimum
environmental
half­
life
of
PFOS
due
to
indirect
photolysis
at
25
º
C
is
greater
than
3.7
years.

Breakdown
products
(
yes/
no):
No
Remarks:
No
evidence
of
direct
or
indirect
photolysis
of
PFOS
was
observed
under
any
of
the
conditions
tested.
Direct
photolytic
decomposition
of
PFOS
was
not
observed
based
on
loss
of
starting
material,
nor
were
any
of
the
predicted
degradation
products
detected
above
their
limits
of
quantitation.

CONCLUSIONS
No
evidence
of
direct
or
indirect
photolysis
of
PFOS
was
observed
under
any
of
the
conditions
tested.
The
mean
and
standard
deviation
of
the
observed
PFOS
concentrations
in
an
aqueous
Fe2O3/
H2O2
matrix
indicate
that
the
indirect
photolytic
half­
life
of
PFOS
at
25
º
C
is
greater
than
3.7
years.

Submitters'
Remarks:
The
authors
assigned
a
Klimisch
ranking
of
2
for
the
data
quality
of
this
study.

Reviewer's
Remarks:
None
REFERENCE
Hatfield,
T.
2001b.
Screening
studies
on
the
aqueous
photolytic
degradation
of
potassium
perfluorooctane
sulfonate
(
PFOS).
3M
Environmental
Laboratory.
3M
Company,
St.
Paul,
MN.
Report
number
W2775.
ENV/
JM/
RD(
2002)
17/
FINAL
252
BIOCONCENTRATION
IN
FISH
Title:
Perfluorooctanesulfonate,
potassium
salt
(
PFOS):
A
flow­
through
bioconcentration
test
with
bluegill
(
Lepomis
macrochirus)

TEST
SUBSTANCE
Identity:
Perfluorooctanesulfonate;
may
also
be
referred
to
as
PFOS
or
FC­
95.
(
Octanesulfonic
acid,
1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8­
heptadecafluoro­,
potassium
salt,
CAS
#
2795­
39­
3)

Remarks:
Sample
from
3M
production
lot
number
217.
The
test
substance
was
a
white
powder.
The
purity
was
determined
to
be
86.9%
by
LC/
MS,
1H­
NMR,
19F­
NMR,
and
elemental
analysis
techniques.

METHODS
Method/
guideline
followed:
USEPA
OPPTS
850.1730
and
OECD
305
Test
type:
Flow­
through
exposure
with
flow­
through
depuration
phase
(
flow
rate:
approximately
6
volume
additions
per
24
hours)

GLP:
Yes
Year
study
performed:
2001
Species:
Bluegill
sunfish
(
Lepomis
macrochirus);
mean
length
=
62
mm
(
56­
66
mm);
mean
weight:
2.70
g
(
2.03­
3.32
g);
age
~
7
months
old
Supplier:
Osage
Catfisheries,
Inc.
Osage
Beach,
Missouri
Concentrations
tested:
0,
0.086,
and
0.87
mg/
L
Uptake
period:
62
days
(
0.086
mg/
L
exposure);
35
days
(
0.087
mg/
L
exposure
 
this
exposure
ended
after
35
days
due
to
fish
mortality)

Depuration
period:
56
days
(
0.086
mg/
L
exposure);
None
(
0.87
mg/
L
exposure)

Analytical
monitoring:
Analyses
of
test
solutions
and
fish
tissues
were
performed
at
Wildlife
International,
Ltd.
Water
samples
were
diluted
and
analyzed
by
HPLC
with
a
single
quadruple
mass
spectrometric
detection.
Tissue
samples
were
homogenized,
extracted,
diluted,
and
analyzed
by
HPLC
with
triple
quadruple
mass
spectrometric
detection.
When
determining
the
concentration
of
the
test
substance
in
the
samples,
the
same
and
most
prominent
peak
response
for
perfluorooctanesulfonate
was
used.
No
attempt
was
made
to
quantify
on
the
basis
of
individual
isomeric
components.
The
LOQ
was
0.05
mg/
L
for
water
in
this
study.
For
tissue
samples,
the
LOQ
was
calculated
on
an
individual
basis
for
each
sample
since
each
entire
submitted
sample,
of
differing
weight,
was
extracted
without
an
adjustment
to
constant
weight.

Recovery
was
excellent
in
both
water
and
fish
tissues,
ranging
from
84.9
to
122%
of
fortification
levels.
Analytical
results
were
not
corrected
for
procedural
recovery.

Collection
of
tissue
samples:
Fish
were
collected
from
test
chambers
by
random
selection
at
12
time
points
during
the
62­
day
uptake
phase.
They
were
euthanized,
blotted
dry,
weighed,
and
measured.
Fish
ENV/
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RD(
2002)
17/
FINAL
253
were
then
rinsed
with
dilution
water,
blotted
dry
again,
and
dissected
into
edible
and
nonedible
fractions.
The
fractions
were
individually
weighed.
The
head,
fins,
and
viscera
were
considered
to
be
nonedible
tissue.
The
remaining
tissue
including
skin
was
considered
to
be
edible
tissue.

Statistical
methods:
Whole
fish
concentrations
were
calculated
based
on
the
sum
of
the
edible
and
nonedible
parts.
Steady­
state
bioconcentration
BCF
values
calculated
from
the
tissue
concentrations
at
apparent
steady­
state
divided
by
the
mean
water
concentration.
Tissue
concentrations
were
considered
to
be
at
apparent
steady­
state
if
3
or
more
consecutive
sets
of
tissue
concentrations
were
not
significantly
different
(
p>
0.05).
Tissue
concentrations
were
evaluated
for
normality
and
homogeneity
of
variance
using
the
Shapiro­
Wilk's
test
and
Bartlett's
test,
respectively.
If
the
data
did
not
meet
the
assumptions,
data
were
transformed
in
an
attempt
to
correct
the
data.
Mean
tissue
concentrations
were
then
compared
using
ANOVA
and
Dunnett's
test.

The
kinetic
bioconcentration
factor
(
BCFK),
uptake
rate
(
k1),
and
depuration
rate
(
k2)
were
calculated
for
the
edible,
nonedible,
and
whole
fish
exposed
to
0.086
mg/
L
PFOS
using
BIOFAC
computer
software.
BIOFAC
is
a
nonlinear
parameter
estimate
routine,
which
estimates
rate
constants
from
a
set
of
sequential
time­
concentration
data.
These
rate
constants
were
then
used
to
calculate
a
BCFK
(
BCFK
=
k1/
k2).

Test
conditions:
Stainless
steel
aquaria
(
104
L)
filled
with
approximately
80
L
solution
were
used
as
exposure
vessels.
One
vessel
per
concentration
was
used.
The
loading
rate
was
0.48
g
fish/
L/
day
(
90
fish
per
vessel).
The
fish
were
fed
flake
food
(
Ziegler
Brothers,
Inc.,
Gardners,
PA)
and
given
16
hours
of
light
and
8
hours
of
dark
with
a
30
minute
transition
period.
The
light
intensity
was
278
lux
at
the
surface
of
the
negative
control
vessel
at
test
initiation.

Two
stock
solutions
were
prepared
at
10
and
100
mg
a.
i./
L.
Stock
solutions
were
stirred
with
an
electric
top­
down
mixer
to
aid
in
the
solubilization
of
the
test
substance.
After
mixing,
the
stocks
appeared
clear
and
colorless.
Stocks
were
prepared
at
approximately
weekly
intervals
during
the
uptake
phase.
Stocks
were
injected
into
the
diluter
mixing
chambers
at
a
rate
of
3.5
mL/
minute
where
they
were
mixed
with
dilution
water
at
a
rate
of
350
mL/
minute
to
achieve
the
desired
test
concentrations.
All
final
test
solutions
appeared
clear
and
colorless.
The
dilution
water
was
moderately­
hard
well
water
with
specific
conductance
of
313
umhos/
cm,
hardness
of
130
mg/
L,
alkalinity
of
178,
and
pH
of
8.1.
During
the
test,
the
dissolved
oxygen
levels
ranged
from
6.4­
8.6
mg/
L,
the
temperature
ranged
from
21.7
to
22.0
º
C,
and
the
pH
ranged
from
7.9
to
8.2.

Remarks:
None
RESULTS
Bioconcentration
factors
(
BCF):

0.086
mg/
L
apparent
steady­
state
BCF:

Edible:
484
Nonedible:
1124
Whole
fish:
856
0.87
mg/
L(
study
ended
prior
to
achieving
steady­
state)
BCF:

Edible:
136
ENV/
JM/
RD(
2002)
17/
FINAL
254
Nonedible:
386
Whole
fish:
278
BIOFAC
Estimates
(
using
0.086
mg/
L
exposure):

Edible
Nonedible
Whole
fish
BCFK:
1866
4312
3614
Time
to
reach
50%
clearance:
146
days
133
days
152
days
PFOS
Concentrations
in
Tissues
of
Bluegill
Exposed
to
0.086
mg/
L:
(
Values
are
from
4
individual
fish
at
each
sample
period.)

Uptake
day
Edible
tissue,
mg/
kg
Nonedible
tissue,
mg/
kg
Whole
fish
conc.,
mg/
kg
0
(
4
hours)
0.167,
0.155,
0.144,
0.182
0.415,
0.519,
0.417,
0.497
0.293,
0.351,
0.286,
0.363
1
0.734,
0.726,
0.631,
0.806
1.68,
1.85,
1.72,
2.07
1.26,
1.34,
1.29,
1.53
3
1.73,
2.07,
2.03,
2.11
4.59,
5.50,
5.47,
5.97
3.21,
4.04,
4.18,
4.38
7
3.73,
4.25,
4.73,
6.25
10.2,
10.6,
11.9,
15.2
7.33,
7.66,
8.73,
11.4
14
11.4,
9.07,
13.7,
12.6
27.3,
23.2,
35.3,
32.6
20.2,
16.9,
26.0,
24.6
21
11.7,
12.0,
12.9,
10.6
33.3,
22.7
24.6,
24.4
23.3,
18.4,
19.8,
18.5
28
18.3,
13.7,
23.9,
23.1
49.4,
40.7,
65.3,
57.9
35.3,
29.2,
45.4,
44.1
35
22.6,
27.7,
23.8,
20.6
67.1,
73.3,
62.0,
59.1
46.3,
53.8,
46.6,
40.9
42
27.6,
25.3,
21.2,
27.6
64.0,
68.1,
54.4,
79.6
50.1,
49.4,
40.9,
56.3
49
33.3,
36.2,
39.0,
30.6
85.0,
95.1,
93.1,
77.7
62.8,
69.6,
70.8,
57.4
56
48.3,
38.9,
44.1,
38.3
122,
94.2,
73.2,
106
90.6,
71.6,
63.3,
74.8
62
42.4,
66.2,
42.2,
39.2
101,
112,
105,
96.4
77.0,
92.7,
79.6,
73.1
Depuration
day
14
48.5,
31.8,
31.6,
42.0
124,
79.4,
81.8,
113
90.3,
60.4,
61.6,
85.3
28
26.0,
33.3,
38.7,
55.8
85.7,
95.1,
85.7,
94.8
58.2,
70.1,
68.1,
81.1
42
24.1,
31.2,
30.0,
33.0
71.7,
80.6,
78.3,
82.1
51.4,
61.4,
61.0,
62.2
56
21.1,
37.6,
32.9,
31.2
57.7,
80.3,
85.4,
84.4
41.6,
66.5,
65.8,
62.1
ENV/
JM/
RD(
2002)
17/
FINAL
255
PFOS
Concentrations
in
Tissues
of
Bluegill
Exposed
to
0.87
mg/
L:
(
Values
are
from
4
individual
fish
at
each
sample
period.)

Uptake
day
Edible
tissue,
mg/
kg
Nonedible
tissue,
mg/
kg
Whole
fish
conc.,
mg/
kg
0
(
4
hours)
1.46,
1.48,
1.19,
1.39
3.52,
4.37,
4.22,
4.06
2.71,
3.08,
2.84,
2.89
1
4.68,
6.59,
5.56,
5.64
11.1,
14.2,
13.3,
12.1
8.00,
10.9,
10.2,
9.47
3
17.3,
15.8,
19.0,
20.8
39.3,
42.0,
43.8,
51.8
30.5,
30.7,
34.5,
39.1
7
42.0,
44.0,
57.7,
46.8
100,
102,
102,
120
74.9,
77.0,
85.3,
89.8
14
87.1,
81.6,
90.7,
73.3
177,
207,
245,
214
141,
157,
180,
158
21
79.4,
117,
104,
102
201,
278,
246,
229
146,
210,
185,
172
281
102,
131,
107,
133
289,
372,
320,
361
205,
267,
232,
263
1Sampling
of
fish
stopped
after
Uptake
day
28
due
to
mortality.

Remarks:
Test
organism
mortality
was
none
in
the
negative
control
(
during
both
the
uptake
and
depuration
phases).
At
0.086
mg/
L,
one
fish
died
after
49
days
and
one
died
after
59
days.
None
died
during
the
depuration
phase.
At
0.87
mg/
L,
mortality
was
first
noted
on
day
9
and
continued
through
day
35
of
the
uptake
phase,
at
which
time
all
fish
either
died
or
had
been
sampled.

Was
control
response
satisfactory:
Yes
Statistical
results:
None
CONCLUSIONS
PFOS
bioconcentrated
in
the
tissues
of
bluegill
sunfish
during
this
study.
Apparent
steady­
state
was
attained
on
Day
49
for
the
fish
exposed
to
0.086
mg
a.
i./
L.
Although
Day
49,
56,
and
62
tissue
residues
were
not
statistically
significantly
different,
PFOS
concentrations
appeared
to
be
still
increasing
during
this
time.
Apparent
steady­
state
BCF
values
for
edible,
nonedible,
and
whole
fish
tissues
were
calculated
to
be
484,
1124,
and
859,
respectively.

PFOS
depurated
slowly.
The
BIOFAC
estimates
for
the
time
to
reach
50%
clearance
for
edible,
nonedible,
and
whole
fish
tissues
were
146,
133,
and
152
days,
respectively.

Submitters'
remarks:
The
authors
assigned
this
study
a
Klimisch
data
reliability
ranking
of
1.

Reviewers'
remarks:
None
REFERENCE
Drottar,
K.,
VanHoven,
R.,
and
H.
Krueger.
2001.
Perfluorooctanesulfonate,
potassium
salt
(
PFOS):
A
flow­
through
bioconcentration
test
with
bluegill
(
Lepomis
macrochirus).
Wildlife
International,
Limited.
Project
number
454A­
134.
3M
Company.
St.
Paul,
MN.
ENV/
JM/
RD(
2002)
17/
FINAL
256
Annex
6.
Robust
Summaries
of
Toxicology
and
Human
Biomonitoring
Studies
PHARMACOKINETIC
STUDIES
Title:
Absorption
of
FC­
95­
14C
in
Rats
after
a
Single
Oral
Dose
(
1979)

TEST
SUBSTANCE
Identity:
Potassium
perfluorooctylsulfonate,
CAS
2795­
39­
3
Remarks:
FC­
95­
14C
(
carbon­
14
label
alpha
to
sulfur
atom,
Riker
Isotope
Inventory
Number
442).
The
specific
activity
is
0.459
+­
0.008
uCi/
mg.
Thin­
layer
and
column
chromatography
showed
the
FC­
95­
14C
to
be
at
least
99%
radiochemically
pure.
The
FC­
95­
14C
was
found
to
be
suitable
for
metabolism
studies.
(
Synthesis
described
in
Johnson
and
Behr,
1979).

METHOD
Method/
guideline
followed:
NA
Test
type:
in
vivo
Species/
strain/
cell
type
or
line:
rat,
Charles
River
CD
Sex:
male
Age
and
body
weight
range
of
animals
used:
8
weeks,
bw
mean
285
g
(
range
243­
315)

Number
of
animals/
sex/
dose:
24
Route
of
administration:
oral
Vehicle:
0.9%
NaCL
solution
containing
1.2
mg
FC­
95­
14C/
2.0
ml
Doses:
4.2
mg/
kg
average,
single
dose
Excretion
routes,
body
fluids,
and
tissues
monitored
and/
or
sampled
during
study:
red
blood
cells,
plasma,
urine,
feces,
spleen,
digestive
tract
plus
contents
(
esophagus,
stomach,
small
intestine,
large
intestine,
and
colon),
and
carcass
Statistical
methods
used:
mean,
log
mean
concentration
versus
time
least
squares
line
Method
remarks:
Rats
were
conditioned
to
individual
metal
metabolism
cages
for
24
hours
prior
to
dosing.
Rats
were
allowed
free
access
to
Purina
Ground
Chow
and
water
before
and
after
dosing.
Each
non­
fasted
rat
was
weighed
immediately
before
being
given
a
single
oral
dose
of
FC­
95­
14C.
The
dosing
solution
was
prepared
by
adding
~
200
mg
of
FC­
95­
14C
to
0.9%
NaCl,
shaking
for
one
half
hour
at
moderate
speed
in
a
mechanical
shaker,
and
centrifuging.
The
supernatant
was
removed
and
used
for
dosing
solution.
The
carbon­
14
content
of
the
dosing
solution
was
determined
by
direct
counting.
The
dose
was
delivered
with
a
2.0
cc
glass
syringe
(
Trylon)
fitted
with
a
stainless
steel
intubation
tube.
Recovery
of
total
carbon­
14
from
series
of
FC­
95­
14C
spiked,
blank
biological
samples
was
used
to
adjust
the
recovery
of
the
test
samples
to
account
for
label
loss
during
the
experimental
manipulations.
ENV/
JM/
RD(
2002)
17/
FINAL
257
Groups
of
three
rats
were
sacrificed
by
exsanguination
at
1,
2,
6,
12,
24,
48,
96,
and
144
hours
post
dose.
Rats
were
anesthetized
with
diethyl
ether
and
blood
was
drawn
from
the
descending
aorta
of
each
rat
and
immediately
transferred
to
a
heparinized
tube.
Plasma
was
prepared
promptly
by
centrifugation.
In
addition
to
plasma
and
red
blood
cells,
total
urine,
total
feces,
spleen,
digestive
tract
plus
contents
(
esophagus,
stomach,
small
intestine,
large
intestine,
and
colon),
and
remainder
of
carcass
were
saved
from
each
of
the
three
rats
in
the
24
and
48
hours
post
dose
groups
for
carbon­
14
analysis.

RESULTS
Detailed
results:
After
a
single
oral
dose
of
FC­
95­
14C
(
mean
dose,
4.2
mg/
kg)
in
solution
to
groups
of
three
male
rats,
at
least
95%
of
the
total
carbon­
14
is
systemically
absorbed
at
24
hours.
The
half­
life
for
elimination
of
total
carbon­
14
from
plasma
is
7.5
days.

The
digestive
tract
and
contents
contained
on
the
average,
3.45%
of
the
dose.
The
mean
fecal
excretion
is
1.55%
of
the
dose
at
24
hours
and
3.24%
at
48
hours.
At
24
hours,
the
mean
sum
of
total
carbon­
14
in
feces
and
digestive
tract
plus
contents
is
5%
of
the
dose.
Some
of
this
5%
likely
represents
systemically
absorbed
carbon­
14
present
either
in
the
digestive
tract
tissues
or
in
the
digestive
tract
contents
as
a
result
of
excretion.
The
data
from
the
48
hour
post
dose
group
of
rats
are
consistent
with
the
24
hour
post
dose
data.
Thus,
at
least
95%
of
the
FC­
95­
14C
dose
was
absorbed
from
solution
after
administration
to
nonfasted
rats.
The
major
portion
of
the
radioactivity
recovered
was
found
in
the
carcass.
The
carcass
data
are
not
as
reliable
as
the
other
tissue
data
since
large
volume
homogenates
were
necessary
and
homogeneity
of
sample
aliquots
was
difficult
to
assure.
There
is
some
excretion
of
total
carbon­
14
in
urine
(
1­
2%/
day).
The
spleens
from
the
24
hour
and
48
hour
post
dose
rats
were
analyzed
for
total
carbon­
14
content,
and
the
percent
of
the
dose
in
the
whole
organ
was
~
0.2%.
The
concentrations
of
total
carbon­
14
in
red
blood
cells
and
plasma
were
compared.
The
mean
ratio
of
red
blood
cell
to
plasma
concentration
at
24
and
48
hours
is
0.25
and
0.39,
respectively.
Thus,
at
24
and
48
hours
after
a
single
oral
dose
of
FC­
95­
14C,
there
is
no
selective
retention
of
carbon­
14
in
red
blood
cells.

The
half­
life
of
elimination
from
plasma
was
determined
by
analysis
of
plasma
samples
from
groups
of
three
rats
at
1,
2,
6,
12,
24,
48,
96,
and
144
hours
after
a
single
oral
dose
of
FC­
95­
14C.
The
log
of
mean
concentration
versus
time
for
these
data
was
plotted.
The
least
squares
line
through
the
individual
points
from
24
to
144
hours
for
these
data
fits
the
equation:
Cp
=
15.65e^(­
0.00387t)
where
Cp
is
plasma
concentration.
The
half­
life
of
elimination
from
plasma
is
179
hours
(
7.5
days).
Thus,
elimination
from
plasma
of
total
carbon­
14
after
a
single
oral
dose
of
FC­
95­
14C
is
slow.

Metabolites
measured:
none
CONCLUSIONS
agree
REFERENCE
Absorption
of
FC­
95­
14C
in
Rats
after
a
Single
Oral
Dose.
Riker
Laboratories,
Inc.,
Subsidiary
of
3M,
St.
Paul,
MN.
Project
No.
890310200.
Johnson,
JD,
Gibson,
SJ,
and
Ober,
RF,
October
26,
1979.
ENV/
JM/
RD(
2002)
17/
FINAL
258
Title:
Extent
and
Route
of
Excretion
and
Tissue
Distribution
of
Total
Carbon­
14
in
Rats
after
a
Single
Intravenous
Dose
of
FC­
95­
14C
(
1979)

TEST
SUBSTANCE
Identity:
Potassium
perfluorooctylsulfonate,
CAS
2795­
39­
3
Remarks:
FC­
95­
14C
(
carbon­
14
label
alpha
to
sulfur
atom,
Riker
Isotope
Inventory
Number
442).
The
specific
activity
is
0.459
+­
0.008
uCi/
mg.
Thin­
layer
and
column
chromatography
showed
the
FC­
95­
14C
to
be
at
least
99%
radiochemically
pure.
The
FC­
95­
14C
was
found
to
be
suitable
for
metabolism
studies.
(
Synthesis
described
in
Johnson
and
Behr,
1979).

METHOD
Method/
guideline
followed:
NA
Test
type:
in
vivo
Species/
strain/
cell
type
or
line:
rat,
Charles
River
CD
Sex:
male
Age
and
body
weight
range
of
animals
used:
8
weeks,
bw
mean
288
g
(
range
262­
303)

Number
of
animals/
sex/
dose:
6
Route
of
administration:
iv,
via
tail
vein
Vehicle:
0.9%
NaCL
solution
containing
1.2
mg
FC­
95­
14C/
2.0
ml
Doses:
4.2
mg/
kg
average,
single
dose
Excretion
routes,
body
fluids,
and
tissues
monitored
and/
or
sampled
during
study:
urine,
feces,
liver,
plasma,
kidney,
lung,
spleen,
bone
marrow,
adrenals,
skin,
testes,
muscle,
fat,
eye,
brain
Statistical
methods
used:
mean,
standard
deviation
Method
remarks:
Rats
were
conditioned
to
individual
metal
metabolism
cages
for
24
hours
prior
to
dosing.
The
rats
were
allowed
free
access
to
Purina
Ground
Chow
and
water
before
and
after
dosing.
Each
rat
was
weighed,
anesthetized
with
diethyl
ether,
then
given
a
single
iv
dose
using
a
3.0
cc
disposable
plastic
syringe
fitted
with
a
26
gauge
1/
2"
needle.
Urine
and
feces
were
collected
at
intervals
for
each
of
the
six
rats
for
89
days.
At
89
days
post
dose,
the
rats
were
anesthetized
with
diethyl
ether;
blood
was
drawn
from
the
descending
aorta,
animals
were
sacrificed
by
exsangination,
and
tissue
samples
were
collected.

RESULTS
Detailed
results:
By
89
days
post
dose,
mean
urinary
excretion
was
30.2+­
1.5%
of
total
C­
14
administered.
Mean
cumulative
fecal
excretion
was
12.6+­
1.2%.
The
authors
note
that
radioactive
content
in
feces
was
too
low
to
measure
after
64
days.
At
day
89,
mean
tissue
C­
14
concentrations
above
one
ug
FC­
95­
14C
ENV/
JM/
RD(
2002)
17/
FINAL
259
equivalents/
g
were
as
follows:
liver,
20.6;
plasma,
2.2;
kidney,
1.1;
and
lung,
1.1.
Other
tissues
such
as
muscle,
skin,
bone
marrow,
and
spleen
had
concentrations
ranging
from
0.2
to
0.6
ug/
g.
There
was
a
difference
in
C­
14
content
of
subcutaneous
fat
(
0.2
ug/
g)
and
abdominal
fat
(<=
0.08
ug/
g).
Very
little
C­
14
was
found
in
whole
eye
(
0.16
ug/
g)
and
no
detectable
C­
14
was
found
in
brain.
Only
liver
and
plasma
contained
a
substantial
percentage
of
dose
at
89
days
post
dose,
25.21%
and
2.81%,
respectively.
The
low
levels
of
radioactivity
found
for
kidney,
lung,
testes,
and
spleen
are
due
in
part
to
blood
still
contained
in
these
organs
when
homogenized.

Mean
Excretion
of
Total
Carbon­
14
in
Urine
Over
Time
Collection
Period
Percent
Dose
(
Days)
During
Period
0­
0.5
0.91
0.5­
1
0.77
1­
2
1.21
2­
3
1.03
3­
4
0.93
4­
5
0.83
5­
6
0.71
6­
7
0.76
7­
8
0.75
8­
9
0.68
9­
10
0.68
10­
11
0.59
11­
12
0.58
12­
13
0.59
13­
14
0.55
14­
15
0.54
15­
16
0.51
16­
17
0.48
17­
18
0.43
18­
19
0.39
19­
21
0.84
21­
23
0.78
23­
25
0.66
25­
27
0.68
27­
29
0.68
29­
32
0.86
32­
36
1.05
36­
40
0.99
40­
43
0.75
43­
47
0.92
47­
50
0.68
50­
54
0.78
54­
57
0.61
57­
61
0.79
61­
69
1.50
69­
78
1.64
78­
89
2.08
Total
30.2
Mean
Excretion
of
Total
Carbon­
14
in
Feces
Over
Time
ENV/
JM/
RD(
2002)
17/
FINAL
260
Collection
Period
Percent
Dose
(
Days)
During
Period
0­
0.5
0.049
0.5­
1
0.842
1­
2
0.795
2­
3
0.649
3­
4
0.656
4­
5
0.577
5­
6
0.510
6­
7
0.588
7­
8
0.482
8­
9
0.421
9­
10
0.387
10­
11
0.370
11­
12
0.296
12­
13
0.310
13­
14
0.281
14­
15
0.276
15­
16
0.272
16­
17
0.187
17­
18
0.163
18­
19
0.129
19­
21
0.311
21­
23
0.302
23­
25
0.262
25­
27
0.208
27­
29
0.202
29­
32
0.223
32­
36
0.526
36­
50
1.530
50­
64*
0.833
Total
12.6
*
The
radioactive
content
of
the
feces
was
too
low
to
measure
after
64
days.

Metabolites
measured:
none.

CONCLUSIONS
agree
REFERENCE
Extent
and
Route
of
Excretion
and
Tissue
Distribution
of
Total
Carbon­
14
in
Rats
after
a
Single
Intravenous
Dose
of
FC­
95­
14
C.
Riker
Laboratories,
Inc.,
Subsidiary
of
3M,
St.
Paul,
MN.
Johnson,
JD,
Gibson,
SJ,
and
Ober,
RE
,
December
28,
1979.
ENV/
JM/
RD(
2002)
17/
FINAL
261
Title:
Cholestyramine­
Enhanced
Fecal
Elimination
of
Carbon­
14
in
Rats
after
Administration
of
Ammonium
[
14C]
Perfluorooctanoate
or
Potassium
[
14C]
Perfluorooctanesulfonate
(
1984)

TEST
SUBSTANCE
Identity:
Potassium
perfluorooctanesulfonate
(
14C­
PFOS)
Ammonium
perfluorooctanoate
(
14C­
PFO)

Remarks:
14C­
PFOS:
sp
act
0.46
uCI/
mg,
radiochemical
purity
>
99%,
14C
label
in
PFOS
is
adjacent
to
sulfur
14C­
PFO:
sp
act
0.51
uCI/
mg,
radiochemical
purity
>
98%

METHOD
Method/
guideline
followed:
NA
Test
type:
in
vivo
Species/
strain/
cell
type
or
line:
rat,
Charles
River
CD
Sex:
male
Age
and
body
weight
range
of
animals
used:
12
weeks,
300­
342
g
Number
of
animals/
sex/
dose:
5
Route
of
administration:
iv
Vehicle:
0.9%
NaCl,
2
ml/
rat
Doses:
Potassium
[
14C]
Perfluorooctanesulfonate
(
PFOS):
3.4
mg/
kg
mean,
single
dose,
0.56
mg/
ml
PFOS
control
animals:
3.5
mg/
kg
mean
Ammonium
[
14C]
Perfluorooctanoate
(
PFO):
13.3
mg/
kg
mean,
single
dose,
2.1
mg/
ml
PFO
control
animals:
13.5
mg/
kg
mean
Excretion
routes,
body
fluids,
and
tissues
monitored
and/
or
sampled
during
study:
Urine,
plasma,
red
blood
cells,
liver
Statistical
methods
used:
mean,
standard
deviation,
Student's
t
test
Method
remarks:
Rats
were
housed
in
individual
stainless­
steel
metabolism
cages
and
fasted
with
free
access
to
water
for
24
hrs
prior
to
receiving
the
fluorochemicals.
The
radiolabeled
compounds
were
administered
as
single
intravenous
doses
(
lateral
tail
vein).
Two
ml
of
dosing
solution
was
administered
to
each
rat.
Ten
rats
were
dosed
with
each
compound.
Five
rats
from
each
group
were
fed
cholestyramine
(
dried
and
ground
resin
Z­
620),
4%
in
feed
(
Purina
Lab
Chow),
for
14
days
after
administration
of
PFO
and
for
21
days
after
administration
of
PFOS.
Control
rats
were
administered
radiolabeled
fluorochemical
but
were
not
treated
with
cholestyramine.
In
order
to
allow
comparison
of
the
radiometric
results
on
an
absolute
basis,
the
ENV/
JM/
RD(
2002)
17/
FINAL
262
radiolabel
doses
were
not
adjusted
for
individual
body
weights.
Urine
and
feces
samples
were
collected
at
intervals
for
individual
rats
in
each
group
until
14
days
after
14C­
PFO
administration
and
21
days
after
14C­
PFOS
administration.
At
these
times,
rats
were
anesthetized
with
diethyl
ether
and
exsanguinated
by
drawing
blood
from
the
descending
aorta.
Plasma
and
red
blood
cells
were
prepared
promptly
by
centrifugation.
Liver
was
collected
as
the
whole
organ
and
stored
frozen
until
analysis.

RESULTS
Detailed
results:
After
21
days
of
cholestyramine
treatment,
the
mean
percentage
of
14C­
PFOS
dose
eliminated
via
feces
(
75.8
+­
5.0)
was
9.5­
fold
the
mean
percentage
of
dose
eliminated
via
feces
by
control
rats
(
8.0
+­
0.8).
After
adjustment
for
the
amount
of
carbon­
14
excreted
in
urine
(
18%
for
controls
and
5%
for
cholestyramine­
treated),
the
amounts
of
carbon­
14
remaining
to
be
excreted
are
19%
for
cholestyraminetreated
rats
and
74%
for
control
rats.
After
14C­
PFOS
administration,
the
mean
liver
carbon­
14
content
at
21
days
represents
11%
and
40%
of
the
dose
for
cholestyramine­
treated
and
control
rats,
respectively.
Mean
plasma
and
red
blood
cell
carbon­
14
concentrations
are
significantly
lower
after
21
days
of
cholestyramine
treatment.

After
14
days
of
cholestyramine
treatment,
the
mean
percentage
of
14C­
PFO
dose
eliminated
via
feces
(
43.2
+­
5.5)
was
9.8­
fold
the
mean
percentage
of
dose
eliminated
via
feces
by
control
rats
(
4.4
+­
1.0).
After
adjustment
for
the
amount
of
carbon­
14
excreted
in
urine
(
67%
for
controls
and
41%
for
cholestyramine­
treated),
the
amounts
of
carbon­
14
remaining
to
be
excreted
are
16%
for
cholestyraminetreated
rats
and
28%
for
control
rats.
After
14C­
PFO
administration,
the
mean
liver
carbon­
14
content
at
14
days
represents
4%
and
8%
of
the
dose
for
cholestyramine­
treated
and
control
rats,
respectively.
Mean
plasma
and
red
blood
cell
carbon­
14
concentrations
are
significantly
lower
after
14
days
of
cholestyramine
treatment.

Carbon­
14
Concentration
(
expressed
as
ug
eq/
g
tissue
or
ml
fluid)
Treatment
Group
Liver
Plasma
Red
Blood
Cells
14C­
PFOS
Cholestyramine
9.4+­
1.6*
0.9+­
0.1*
0.3+­
0.1*
Control
35.6+­
5.6
6.9+­
0.6
1.8+­
0.4
14C­
PFO
Cholestyramine
12.1+­
2.1*
5.1+­
1.7*
1.8+­
0.7*
Control
22.3+­
6.2
14.7+­
6.8
4.2+­
2.4
*
Significantly
different
from
control
values
(
p<
0.05)

The
authors
conclude
that
the
high
concentration
of
14C­
PFOS
or
14C­
PFO
in
liver
at
2
to
3
weeks
after
dosing
and
the
fact
that
cholestyramine
treatment
enhances
fecal
elimination
of
carbon­
14
by
nearly
10­
fold
suggest
that
there
is
a
considerable
enterohepatic
circulation
of
14C­
PFOS
and
14C­
PFO.

Metabolites
measured:
none
CONCLUSIONS
agree
ENV/
JM/
RD(
2002)
17/
FINAL
263
REFERENCE
Johnson,
J.
D.,
Gibson,
SJ,
and
Ober,
RE
(
1984).
Cholestyramine­
Enhanced
Fecal
Elimination
of
Carbon­
14
in
Rats
after
Administration
of
Ammonium
[
14C]
Perfluorooctanoate
or
Potassium
[
14C]
Perfluorooctanesulfonate.
Fundamental
and
Applied
Toxicology
4,
pages
972­
976.

See
also
Johnson,
J.
D.,
Gibson,
SJ,
and
Ober
RE
(
1984).
Enhanced
elimination
of
FC­
95­
14C
and
FC­
143­
14C
in
rats
with
cholestyramine
treatment.
Project
No.
8900310200,
Riker
Laboratories,
Inc.
St.
Paul,
MN.
ENV/
JM/
RD(
2002)
17/
FINAL
264
Title:
Oral
(
Gavage)
Pharmacokinetic
Study
of
PFOS
in
Rats,
Analytical
Laboratory
Report,
Determination
of
the
Presence
and
Concentration
of
Perfluorooctanesulfonate
(
PFOS)
in
Serum,
Liver,
Urine,
and
Feces
Samples
TEST
SUBSTANCE
Identity:
Perfluorooctylsulfonate,
potassium
salt,
CAS
2795­
39­
3
Remarks:
Purity
86.9%,
Lot
#
217
METHOD
Method/
guideline
followed:
This
study
was
conducted
in
compliance
with
United
States
Food
and
Drug
Administration
(
FDA)
Good
Laboratory
Practice
(
GLP)
Regulations
21
CFR
Part
58,
with
the
exceptions
noted
on
page
3
of
report.

Test
type:
in
vivo
Species/
strain/
cell
type
or
line:
rat/
Sprague­
Dawley/
pregnant
Crl:
CD(
R)
BR
VAF/
Plus(
R)

Sex:
F0:
female,
F1:
both
Age
and
body
weight
range
of
animals
used:
60
days,
200­
225
g
Number
of
animals/
sex/
dose:
F0:
16,
F1:
5
male
and
5
female
pups/
litter
Route
of
administration:
oral
Vehicle:
0.5%
Tween(
R)
80
in
R.
O.
deionized
water,
dosage
volume
5
ml/
kg
Doses:
0
(
vehicle),
0.1,
0.4,
1.6,
and
3.2
mg/
kg/
day
in
volume
of
5
ml/
kg,
once
daily
beginning
42
days
prior
to
cohabitation,
and
continued
through
day
14
or
day
20
of
presumed
gestation.
Only
the
F0
females
were
dosed.

Excretion
routes,
body
fluids,
and
tissues
monitored
and/
or
sampled
during
study:
F0
urine,
feces,
serum,
liver.
F1
liver
and
serum.

Statistical
methods
used:
mean
and
standard
deviation.

Method
remarks:
Serum,
urine,
and
feces
specimens
were
collected
from
adult
female
rats
(
F0
dams)
before
mating
and
at
gestation
day
(
GD)
7,
GD
15
and
GD
21.
Liver
specimens
were
collected
from
F0
dams
at
termination
of
the
study
(
GD
21).
A
total
of
54
pooled
serum
and
liver
specimens
were
collected
from
fetuses
on
GD
21.
Specimens
were
sent
to
the
3M
Environmental
Laboratory
and
the
contract
labs
to
be
analyzed
for
PFOS.
ENV/
JM/
RD(
2002)
17/
FINAL
265
RESULTS
Detailed
results:
Average
Results
for
the
Analysis
of
Serum
Samples
(
ug/
ml)
0
mg/
kg
0.1
mg/
kg
0.4
mg/
kg
1.6
mg/
kg
3.2
mg/
kg
Day
0
dam
0.0723
8.89
40.7
160
318
Day
7
dam
0.126
7.82
40.9
154
105
Day
15
dam
0.0926
8.80
41.4
156
275
Day
21
dam
0.0714
4.24
26.2
136
155
Day
21
Fetal
0.125
9.07
34.3
101
165
Average
Results
for
the
Analysis
of
Liver
Samples
(
GD
21)
Dose
Group
PFOS
Conc.
(
ug/
g)
PFOS
Conc.
(
ug/
g)
(
mg/
kg/
day)
Female
Adult
(
F0)
Fetal
Liver
(
F1)
0
0.288
0.169
0.1
29.2
7.93
0.4
107
30.6
1.6
347
86.7
3.2
610
230
Average
Results
for
the
Analysis
of
Urine
Samples
from
F0
Dams
0
mg/
kg
0.1
mg/
kg
0.4
mg/
kg
1.6
mg/
kg
3.2
mg/
kg
Day
0
<
LOQ
0.0497
0.302
0.959
1.53
Day
7
<
LOQ
0.0620
0.308
1.10
1.60
Day
15
0.00905
0.0685
0.526
0.622
0.563
Day
21
0.0194
0.0574
0.555
2.71
1.61
Average
Results
for
the
Analysis
of
Feces
Samples
from
F0
Dams
0
mg/
kg
0.1
mg/
kg
0.4
mg/
kg
1.6
mg/
kg
3.2
mg/
kg
Day
0
0.0380
0.499
2.42
10.3
23.9
Day
7
0.0155
0.490
2.16
9.19
33.0
Day
15
0.0322
0.662
2.93
11.1
29.5
Day
21
0.0342
0.416
2.39
9.94
20.1
In
general,
there
was
a
dose­
related
increase
in
the
levels
of
PFOS
in
the
liver
and
serum
of
the
dams
and
the
fetuses.
PFOS
was
also
observed
in
the
control
dams,
as
well
as
the
control
fetuses.
On
sacrifice
on
GD21,
the
levels
of
PFOS
were
much
higher
in
the
liver
than
in
the
serum
for
the
dams.
The
levels
of
PFOS
remained
fairly
steady
in
the
serum
of
the
dams
from
GD0
 
GD
15,
but
the
levels
dropped
at
GD21.
In
the
GD21
fetuses,
the
level
of
PFOS
in
the
serum
was
generally
comparable
to
the
level
observed
in
the
dams,
whereas
the
level
of
PFOS
in
the
fetal
livers
was
well
below
that
seen
in
the
dams.

Results
Remarks:
As
stated
in
the
report.
"
It
is
not
possible
to
verify
true
recovery
of
endogenous
analyte
from
tissues
without
radio­
labeled
reference
material.
The
only
measurement
of
accuracy
available
at
this
time,
matrix
spike
studies,
indicate
that
the
data
are
quantitative
to
50%
or
greater."

Metabolites
measured:
none
ENV/
JM/
RD(
2002)
17/
FINAL
266
CONCLUSIONS
Conclusions:
Under
the
conditions
of
the
present
studies,
PFOS
was
observed
in
the
livers,
urine,
feces
and
sera
of
all
female
rats
dosed
with
PFOS
during
the
in­
life
phase
of
the
study.
Additionally,
PFOS
was
observed
in
fetal
liver
and
serum
taken
during
gestation
from
the
same
group
of
female
rats.

Agree
REFERENCE
Study
Title:
Oral
(
Gavage)
Pharmacokinetic
Study
of
PFOS
in
Rats
Analytical
Laboratory
Report
Title:
Determination
of
the
Presence
and
Concentration
of
Perfluorooctanesulfonate
(
PFOS)
in
Serum,
Liver,
Urine,
and
Feces
Samples
3M
Medical
Department
Study:
T­
6295.12,
Argus
In­
Life
Study:
#
418­
013,
FACT
TOX­
110,
3M
Laboratory
Request
No.
U2849,
3M
Environmental
Laboratory,
May
4,
2001.
ENV/
JM/
RD(
2002)
17/
FINAL
267
Title:
Oral
(
Gavage)
Pharmacokinetic
Recovery
Study
of
PFOS
in
Rats,
Analytical
Laboratory
Report,
Determination
of
the
Concentration
of
Perfluorooctanesulfonate
(
PFOS)
in
the
Serum,
Liver,
Urine,
and
Feces
of
Crl:
CDBR
VAF/
Plus
®
Rats
Exposed
to
PFOS
via
Gavage
TEST
SUBSTANCE
Identity:
Perfluorooctylsulfonate,
potassium
salt
(
FC­
95),
CAS
2795­
39­
3
Remarks:
Purity
86.9%,
Lot
#
217
METHOD
Method/
guideline
followed:
This
study
was
conducted
in
compliance
with
United
States
Food
and
Drug
Administration
(
FDA)
Good
Laboratory
Practice
(
GLP)
Regulations
21
CFR
Part
58,
with
the
exceptions
noted
on
page
3
of
report.
It
does
not
appear
these
exceptions
would
significantly
impact
the
results
or
conclusions.
The
analytical
phase
completed
at
the
3M
Environmental
Laboratory
was
performed
in
accordance
with
3M
Environmental
Technology
and
Safety
Services
Standard
Operating
Procedures.

Test
type:
in
vivo
Species/
strain/
cell
type
or
line:
rat/
Sprague­
Dawley/
pregnant
Crl:
CD(
R)
BR
VAF/
Plus
®
Sex:
F0:
female,
F1:
both
Age
and
body
weight
range
of
animals
used:
65
days,
192­
231
g
Number
of
animals/
sex/
dose:
F0:
8,
F1:
5
male
and
5
female
pups/
litter
Route
of
administration:
oral
Vehicle:
0.5%
Tween(
R)
80
in
R.
O.
deionized
water,
dosage
volume
5
ml/
kg
Doses:
0
(
vehicle),
0.1
and
1.6
mg/
kg/
day
in
volume
of
5
ml/
kg
(
0.00,
0.02,
and
0.32
mg/
mL),
once
daily
beginning
43
days
prior
to
cohabitation
until
confirmed
evidence
of
mating.
Only
the
F0
females
were
dosed.

Excretion
routes,
body
fluids,
and
tissues
monitored
and/
or
sampled
during
study:
F0
urine,
feces,
serum,
liver.
F1
liver
and
serum.

Statistical
methods
used:
means,
standard
deviations,
and
percentages
Method
remarks:
Rat
dams
were
exposed
to
PFOS
via
gavage
prior
to
and
during
mating.
Exposure
to
PFOS
was
halted
on
the
first
day
of
presumed
gestation.
Pups
were
not
directly
exposed
to
PFOS,
but
may
have
been
exposed
in
utero
and
during
lactation.
Male
rats
of
the
same
source
and
strain
were
used
only
as
breeders
and
were
not
administered
the
test
article
or
considered
part
of
the
test
system.

Urine
and
fecal
samples
were
collected
from
F0
female
rats
for
the
following
intervals:
one
day
prior
to
initiation
of
cohabitation
to
the
following
morning,
days
6
to
7,
14
to
15,
and
20
to
21
of
presumed
ENV/
JM/
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2002)
17/
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268
gestation
(
DG
6
to
7,
14
to
15,
and
20
to
21),
and
days
of
lactation
(
DL)
21
to
22.
Blood
samples
were
collected
from
each
of
the
maternal
rats
on
the
day
cohabitation
was
initiated
(
prior
to
cohabitation),
DG
7,
15
and
21,
and
DL
14
and
22.
Day
1
of
lactation
was
defined
as
the
day
of
birth.
On
DL
4,
litters
were
culled
to
five
male
pups
and
five
female
pups
per
litter,
where
possible.
Sera
specimens
were
collected
from
pooled
litter
samples
on
DL
21.
On
DL
22,
all
surviving
Generation
F0
and
Generation
F1
animals
assigned
to
the
study
were
sacrificed,
and
a
liver
specimen
was
collected
from
each
animal.
The
liver
from
each
pup
was
collected
and
pooled
per
litter.
Blood
samples
were
collected
and
pooled
per
liter.
Urine,
fecal,
serum
and
liver
samples
were
shipped
to
the
Sponsor
for
analysis.
On
days
1
to
4
of
the
43­
day
premating
period,
F0
female
rats
received
25%
greater
dose
due
to
an
incorrect
calculated
amount
of
test
substance
in
vehicle.

RESULTS
Detailed
results:

Average
Results
for
the
Analysis
of
Sera
Samples
(
PFOS
Conc.
ug/
ml)
DG0
DG7
DG15
DG21
DL14
DL21
DL22
0.0
mg/
kg
F0
0.100
0.0796
0.0742
<
LLQ
0.0542
NS
0.0492
0.0
mg/
kg
F1
NS
NS
NS
NS
NS
0.0531
NS
0.1
mg/
kg
F0
9.21
7.24
5.68
2.58
1.63
NS
0.979
0.1
mg/
kg
F1
NS
NS
NS
NS
NS
1.80
NS
1.6
mg/
kg
F0
161
129
90.6
39.5
20.6
NS
14.1
1.6
mg/
kg
F1
NS
NS
NS
NS
NS
27.1
NS
Average
Results
for
the
Analysis
of
Liver
Samples
(
PFOS
Conc.
ug/
g)
DL22
0.0
mg/
kg
F0
0.243
0.0
mg/
kg
F1
0.174
0.1
mg/
kg
F0
6.15
0.1
mg/
kg
F1
5.00
1.6
mg/
kg
F0
59.7
1.6
mg/
kg
F1
56.2
Average
Results
for
the
Analysis
of
Urine
Samples
(
PFOS
Conc.
ug/
ml)
DG0
DG6/
7
DG14/
15
DG20/
21
DL21/
22
0.0
mg/
kg
F0
0.00819
0.0100
0.00685
0.00614
<
LOQ
0.1
mg/
kg
F0
0.0905
0.0307
0.0327
0.0231
0.00555
1.6
mg/
kg
F0
2.11
0.888
0.613
0.340
0.0334
Average
Results
for
the
Analysis
of
Feces
Samples
(
PFOS
Conc.
ug/
g)
DG0
DG6/
7
DG14/
15
DG20/
21
DL21/
22
0.0
mg/
kg
F0
ND
ND
ND
ND
ND
0.1
mg/
kg
F0
0.601
0.399
0.294
0.119
0.0522
1.6
mg/
kg
F0
10.9
8.39
4.83
2.06
0.387
In
general,
there
was
a
dose­
related
increase
in
the
levels
of
PFOS
in
the
liver
and
serum
of
the
dams
and
the
levels
in
the
serum
decreased
with
time.
The
levels
of
PFOS
were
much
higher
in
the
liver
than
in
the
serum
of
the
dams
and
the
pups.
The
levels
of
PFOS
were
similar
in
the
liver
of
the
dams
and
pups,
while
the
levels
in
the
serum
were
slightly
higher
in
the
pups
than
in
the
dams.
ENV/
JM/
RD(
2002)
17/
FINAL
269
Results
Remarks:
As
stated
in
the
report.
"
It
is
not
possible
to
verify
true
recovery
of
endogenous
analyte
from
tissues
without
radio­
labeled
reference
material.
The
only
measurement
of
accuracy
available
at
this
time,
matrix
spike
studies,
indicate
that
the
data
are
quantitative
to
50%
or
greater."

Metabolites
measured:
none
CONCLUSIONS
Statement
of
Conclusion
Under
the
conditions
of
the
present
study,
perfluorooctanesulfonate
was
observed
in
all
sample
types
of
all
Generation
0
test
system
animals
dosed
with
the
test
substance
during
the
in­
life
phase
of
the
study,
and
in
all
sample
types
of
their
offspring
(
Generation
F1).

Agree
REFERENCE
Study
Title:
Oral
(
Gavage)
Pharmacokinetic
Recovery
Study
of
PFOS
in
Rats
Analytical
Laboratory
Report
Title:
Determination
of
the
Concentration
of
Perfluorooctanesulfonate
(
PFOS)
in
the
Serum,
Liver,
Urine,
and
Feces
of
Crl:
CDBR
VAF/
Plus
®
Rats
Exposed
to
PFOS
via
Gavage,
3M
Environmental
Laboratory
Report
No.
FACT
TOX­
111,
Laboratory
Request
No.
U2994,
3M
Ref.
No.
T­
6295.14,
Argus
In­
Life
Study
418­
015,
May
4,
2001.
ENV/
JM/
RD(
2002)
17/
FINAL
270
Title:
Half­
Life
Study
of
PFOS
in
Serum,
2000
TEST
SUBSTANCE
Identity:
PFOS
METHOD
Study
design:
PFOS
half­
life
study
on
retired
workers
from
the
Decatur,
Alabama
plant
Manufacturing/
Processing/
Use:
N/
A
Hypothesis
tested:
To
determine
the
half­
life
of
PFOS
in
retired
perfluorochemical
production
workers.

Study
period:
Nov.
1998
to
2003
Setting:
N/
A
Total
population:
27
retirees
from
the
Decatur,
Alabama
and
Cottage
Grove,
Minnesota
plants
Subject
selection
criteria:
The
participants
volunteered
for
this
study
Total
#
of
subjects
in
study:
18
males
Comparison
population:
N/
A
Participation
rate:
Unknown
Subject
description:
Retirees
ranged
in
age
from
55­
74
years,
worked
in
the
plant
for
an
average
of
28
years,
average
time
from
retirement
to
start
of
study
was
30
months
(
range:
5­
130
months).

Health
effects
studied:
N/
A
Data
collection
methods:
Blood
sera
samples
collected
every
6
months
Details
on
data
collection:
No
information
was
provided
as
to
how
the
blood
was
drawn,
stored,
etc.

Exposure
period:
Unknown.

Description/
delineation
of
exposure
groups/
categories:
N/
A
Measured
or
estimated
exposure:
N/
A
Exposure
levels:
N/
A
Statistical
methods:
Medians
and
ranges
calculated.

Other
methodological
information:
Half­
lives
were
calculated
assuming
a
one­
compartment
model.
A
log­
linear
relationship
was
used
to
estimate
the
serum
fluorochemical
elimination
half­
life
in
participating
ENV/
JM/
RD(
2002)
17/
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271
retirees.
In
this
log
linear
relationship,
the
slope
of
the
line
is
related
to
the
elimination
constant
via
the
equation
slope
=
­
kel(
2.303).
Once
the
elimination
constant
is
calculated,
the
half­
life
is
determined
using
the
relationship
t1/
2
=
0.693/
kel.

If
3
data
points
were
not
available
for
any
of
the
subjects
and
if
there
was
a
lack
of
fit
to
the
model,
that
retiree
was
not
included
in
the
analyses.
Eighteen
participants
met
these
requirements.

RESULTS
Describe
results:
The
median
serum
half­
life
of
PFOS
was
270
days,
with
a
range
of
139
to
640
days.
It
should
be
noted
that
the
difference
in
serum
PFOS
levels
between
retirees
was
quite
large
(
0.2
­
2.0
ppm).

Study
strengths
and
weaknesses:
For
most
of
the
participants
not
included
in
the
analysis,
the
second
measurement
was
higher
than
the
first.
Therefore,
the
data
did
not
fit
the
model
and
they
were
excluded.
Although
this
may
justify
not
including
those
participants
in
the
analysis,
it
is
an
indication
of
the
many
limitations
of
the
data.
It
is
stated
in
the
report
that
neither
age
nor
number
of
months
retired
was
associated
with
the
serum
PFOS
half­
life
calculations;
however,
this
statement
is
not
supported
with
any
data
in
the
report.
In
addition,
no
individual
data
were
provided
in
the
report
and
the
relationship
between
number
of
years
exposed
in
the
workplace
and
PFOS
levels
and
half­
life
were
excluded.
Retirees
were
excluded
from
these
calculations
if
their
second
measurement
was
higher
than
the
first.
It
is
unknown
why
this
occurred,
but
the
exclusion
of
those
retirees
introduces
bias
to
the
results.
Also,
elimination
of
PFOS
occurs
via
urine
and
feces;
however,
these
measurements
were
not
taken.
Therefore,
it
cannot
be
determined
that
the
half­
life
suggested
by
the
preliminary
results
reported
here
represents
a
true
elimination
half­
life
from
the
body.
Finally,
the
effect
of
continued
non­
occupational,
low­
level
exposure
on
the
halflife
is
unknown.

Research
sponsors:
3M
Environmental
Lab
Consistency
of
results:
These
results
are
not
consistent
with
the
first
study
in
which
3
retirees
were
followed
over
a
5­
year
period.
In
that
study,
the
estimated
half­
life
was
3
to
4
years.
There
is
no
explanation
in
this
current
report
as
to
why
there
is
such
a
large
difference
in
the
results
or
why
none
of
the
retirees
in
the
current
study
didn't
fall
within
the
range
of
the
first
study.

CONCLUSIONS
N/
A
REFERENCE
Determination
of
Serum
Half­
Lives
of
Several
Fluorochemicals,
June
8,
2000,
3M
Company.
FYI­
0700­
1378,
8(
e)
Supplemental
Submission,
8EHQ­
0373/
0374.
ENV/
JM/
RD(
2002)
17/
FINAL
272
ACUTE
TOXICITY
STUDIES
Title:
An
Acute
Inhalation
Toxicity
Study
of
T­
2306
CoC
in
the
Rat.
1979
TEST
SUBSTANCE
Identity:
Potassium
perfluorooctylsulfonate,
CAS
No.:
2795­
39­
3
Remarks:
Dust,
PFOS
(
T­
2306
CoC).
Purity
not
specified.

METHOD
Method/
guideline
followed:
Similar
to
OECD
403
GLP:
N,
no
QA/
QC
indicated
Year
study
performed:
1979
Species/
Strain:
Rat/
Sprague­
Dawley
Sex
(
Males/
females/
both):
Both
No.
of
animals/
sex/
dose:
5/
sex/
group
Route
of
Administration:
Inhalation
Remarks:
Concentrations
of
1.89,
2.86,
4.88,
6.49,
7.05,
13.9,
24.09,
45.97
mg/
l
PFOS
were
administered
to
eight
test
groups.
A
Wright
dust­
feed
mechanism
with
dry
air
at
a
flow
rate
of
12
to
16
liters
per
minute
was
used
to
administer
the
PFOS
dust.
Rats
were
exposed
for
1
hour.
The
test
group
rats
weighed
201­
299
g
at
study
initiation.
The
control
group
rats
weighed
203­
263
g
at
study
initiation.
The
control
rats
were
exposed
to
dry
air
at
a
flow
rate
of
12
liters
per
minute.
All
other
protocols
were
the
same
as
the
test
group
rats.
The
rats
were
observed
for
abnormal
signs
prior
to
exposure,
at
15­
minute
intervals
during
the
1­
hour
exposure,
at
removal
from
the
exposure
chamber,
hourly
for
four
hours
after
exposure,
and
daily
thereafter
for
14
days.
Individual
bodyweights
were
recorded
on
Day
0
(
prior
to
exposure),
Day
1,
Day
2,
Day
4,
Day
7,
and
Day
14.
It
is
reported
that
all
animals
dying
spontaneously
were
necropsied
as
soon
as
possible
after
death.
Blood
samples
were
collected
on
Day
14
from
all
surviving
animals,
but
analyses
were
not
provided.

RESULTS
LC50
=
5.2
(
4.4
 
6.4)
mg/
l,
(
95%
confidence
limits);
referenced
method
of
Litchfield
and
Wilcoxon
Number
of
deaths
at
each
dose
level
(
by
sex):

0.0
mg/
l:
0/
10;
1.89
mg/
l:
0/
10;
2.06
mg/
l:
1/
10;
4.88
mg/
l:
2/
10;
6.49
mg/
l:
8/
10;
7.05
mg/
l:
8/
10;
24.09
mg/
l:
10/
10
(
authors
did
not
provide
summary
by
sex)

Remarks:
The
highest
dose
group,
45.97
mg/
l,
was
not
used
in
the
LC50
calculations
and
terminated
on
Day
2.
At
that
point,
only
5
animals
survived
and
blood
samples
were
taken
at
termination.
The
13.9
mg/
l
group
was
also
terminated
early
(
Day
1)
because
of
a
mechanical
problem
during
exposure.
These
animals
ENV/
JM/
RD(
2002)
17/
FINAL
273
were
also
not
used
in
the
LC50
determination.

In
the
24.09
mg/
l
exposure
group,
all
animals
died
by
Day
6.
At
7.05
and
6.49
mg/
l
there
was
80%
mortality
with
last
deaths
at
Day
10.
At
4.88,
2.86,
and
1.89
mg/
l
there
was
20%,
10%,
and
0%
mortality,
respectively.
At
2.86
mg/
l,
deaths
occurred
on
Day
7
and
10.
At
1.89
mg/
l,
one
death
occurred
on
Day
12.
The
rats
in
all
these
groups
showed
signs
of
toxicity
including
emaciation,
red
material
around
the
nose
or
other
nasal
discharge,
yellow
material
around
the
anogenital
region,
dry
rales
or
other
breathing
disturbances,
and
general
poor
condition.
Abnormal
in­
life
observations
were
reported
to
be
less
frequent
in
the
lower
exposure
groups.

The
most
common
abnormality
was
discoloration
of
the
liver
and
lung.
Discoloration
of
the
lung
was
also
observed
in
control
rats
and
therefore
may
not
be
treatment
related.
Therefore,
the
most
significant
treatment­
related
abnormality
was
varying
degrees
of
discoloration
of
the
liver.
Among
animals
that
died
prematurely,
decreased
body
weight,
discoloration
of
the
lung,
and
discoloration
and
distention
of
the
small
intestine
were
also
observed.

CONCLUSIONS
LC50
=
5.2
(
4.4
 
6.4)
mg/
l,
(
95%
confidence
limits).
Only
conclusion
provided;
seems
reasonable
with
available
data
REFERENCE
Rusch,
G.
M.,
W.
E.
Rinehart
and
C.
A.
Bozak.
1979.
An
Acute
Inhalation
Toxicity
Study
of
T­
2306
CoC
in
the
Rat.
Project
No.
78­
7185,
Bio/
dynamics
Inc.
ENV/
JM/
RD(
2002)
17/
FINAL
274
Title:
Fluorad
Fluorochemical
Surfactant
FC­
95
Acute
Oral
Toxicity
(
LD50)
Study
in
Rats.
1978.

TEST
SUBSTANCE
Identity:
Potassium
perfluorooctylsulfonate,
CAS
No.:
2795­
39­
3
Remarks:
FC­
95.
Purity
not
specified.

METHOD
Method/
guideline
followed:
Similar
to
OECD
401
GLP
(
Y/
N):
N,
no
QA/
QC
indicated
Year
study
performed:
1978
Species/
Strain:
Rat/
Charles
River
CD
Sex
(
Males/
females/
both):
both
Number
of
animals/
sex/
dose:
5/
sex/
dose
Vehicle:
20%
acetone/
80%
corn
oil
Route
of
Administration:
gavage
Remarks:
Levels
of
100,
215,
464,
and
1000
mg/
kg
PFOS
were
tested.
All
dose
levels
were
administered
as
volumes
of
10ml/
kg
body
weight.
The
rats
weighed
172­
212
g
at
the
beginning
of
the
study
immediately
prior
to
dosing
and
weights
were
recorded
at
Day
7
and
Day
14.
The
rats
were
observed
for
abnormal
signs
during
the
four
hours
after
exposure,
and
daily
thereafter
for
14
days.
It
is
reported
that
all
animals
dying
spontaneously
were
grossly
necropsied,
as
well
as
all
rats
that
survived
to
the
end
of
the
14­
day
study.

RESULTS
LD50:
251
(
199­
318)
mg/
kg,
(
95%
confidence
limits);
3
references
for
statistical
tables
are
given.

Number
of
deaths
at
each
dose
level
(
by
sex):
100
mg/
kg:
0/
5
males,
0/
5
females;
215
mg/
kg:
2/
5
males,
1/
5
females;
464
mg/
kg:
5/
5
males,
5/
5
females;
1000
mg/
kg:
5/
5
males,
5/
5
females
Remarks:
All
rats
in
the
464
and
1000
mg/
kg
dose
groups
died
before
the
end
of
the
study.
Three
animals
in
the
215
mg/
kg
group
died
prematurely.
It
appears
signs
of
toxicity
most
frequently
observed
included:
hypoactivity,
decreased
limb
tone,
and
ataxia.
At
necropsy
observations
included:
yellowstained
urogenital
region,
stomach
distention
and
signs
of
irritation
of
the
glandular
mucosa,
and
lung
congestion.
No
differences
between
sexes
were
noted.

LD50
male
rats:
233
(
160­
339)
mg/
kg
(
95%
confidence
limits)
LD50
female
rats:
271
(
200­
369)
mg/
kg
(
95%
confidence
limits)
ENV/
JM/
RD(
2002)
17/
FINAL
275
CONCLUSIONS
None
specified
beyond
LD50
REFERENCE
Dean,
W.
P.,
D.
C.
Jessup,
G.
Thompson,
G.
Romig,
and
D.
Powell.
1978.
Fluorad
Fluorochemical
Surfactant
FC­
95
Acute
Oral
Toxicity
(
LD50)
Study
in
Rats.
Study
No.
137­
083,
International
Research
and
Development
Corporation.
(
Includes
Acute
Oral
Toxicity
Study
in
Rats
with
T­
2297
CoC.
Project
No.
78­
1433A,
Biosearch,
Inc.)
.
ENV/
JM/
RD(
2002)
17/
FINAL
276
SKIN
IRRITATION
Title:
Eye
and
Skin
Irritation
Report
on
Sample
T­
1117.
1974
TEST
SUBSTANCE
Identity:
Potassium
perfluorooctylsulfonate,
CAS
No.:
2795­
39­
3
Remarks:
FC­
95,
Sample
T­
1117.
Purity
not
specified.

METHOD
Note
pH
of
test
material:
Not
specified
Method/
Guideline
followed:
Not
specified
Test
Type:







Species/
strain/
cell
type:
Rabbits/
albino
Sex
(
males/
females/
both):
Not
specified
Number
of
animals/
sex/
dose:
6
total
Total
dose:
appears
to
be
1.0
gram,
0.5g
placed
on
each
of
2
prepared
test
sites
(
intact­
wet,
abradedwet
total
dose
not
specified
Vehicle:
None?
Not
specified
Length
of
time
test
material
is
in
contact
with
animal/
cell:
72
hr
Grading
scale:
Separate
scores
for
erythema
formation
and
edema
formation
are
summed.
Reference
source
not
provided.

Remarks:
Six
albino
rabbits
had
their
hair
clipped
from
their
backs
and
flanks,
and
five
tenths
of
one
gram
(
0.5
g)
of
test
material
was
placed
on
abraded­
wet
or
intact­
wet
prepared
test
sites,
then
covered
with
gauze
patches.
After
24
hours
and
72
hours
the
coverings
were
removed
and
the
degree
of
erythema
and
edema
was
recorded
according
to
a
standardized
scale.

RESULTS
Results:
In
all
cases
it
is
reported
the
primary
skin
irritation
scores
were
0;
which
indicates
no
reddening
or
swelling
detected.
Primary
irritation
score:
zero
Remarks:
No
indication
of
reliability.
No
QA/
QC.
No
effects
reported.

CONCLUSIONS
No
irritation.
Inadequate
information
is
presented
in
report
to
evaluate
quality
of
study
and
validity
of
conclusion.

REFERENCE
J.
A.
Biesemeier
and
D.
L.
Harris.
1974.
Eye
and
Skin
Iritation
Report
on
Sample
T­
1117.
Project
No.
4102871,
WARF
Institute
Inc.
ENV/
JM/
RD(
2002)
17/
FINAL
277
EYE
IRRITATION
Title:
Eye
and
Skin
Irritation
Report
on
Sample
T­
1117.
1974
TEST
SUBSTANCE
Identity:
Potassium
perfluorooctylsulfonate,
CAS
No.:
2795­
39­
3
Remarks:
T­
1117.
Purity
not
specified.

METHOD
Note
pH
of
test
material:
Not
specified
Method/
Guideline
followed:
Not
specified
Test
Type:
in
vivo
Species/
strain/
cell
type
or
line:
Rabbit/
New
Zealand
White
Sex
(
males/
females/
both):
Not
specified
Number
of
animals/
sex/
dose:
6/
single
dose
Total
dose:
it
appears
0.1
gram,
(
protocol
states
either
0.1
ml
or
0.1g
and
T­
1117
is
reported
to
be
a
solid
{
FC­
95}
by
3M),
total
dose
not
specified
Length
of
time
test
material
is
in
contact
with
animal/
cell:
could
be
72
hours,
But
at
1
hr
observation
and
at
each
scoring
after
"
any
accumulated
discharge
or
residue
of
test
material
was
flushed
from
the
eye."

Observation
period:
1hr,
24
hr,
48
hr,
72
hr
Scoring
method
used:
Not
specified
or
referenced
­
"
The
reaction
to
the
test
material
was
read
according
to
the
scale
of
scoring
for
damage
to
the
cornea,
iris,
and
the
bulbar
and
palpebral
conjunctivae "

Remarks:
Rabbits
were
placed
in
collars
so
they
could
not
rub
their
eyes.
One
tenth
of
a
gram
(
0.1
g)
of
the
test
substance
was
instilled
in
one
eye,
the
other
eye
was
left
untreated
as
a
control.
It
is
reported
that
the
reaction
to
the
test
material
was
read
against
a
scale
of
damage
to
the
cornea,
iris,
and
the
bulbar
and
palpebral
conjunctivae
at
1,
24,
48,
and
72
hours
after
treatment.
The
scale
criteria
are
not
presented
or
referenced.
Each
time
the
eyes
were
scored,
any
accumulated
discharge
or
residue
of
test
material
was
flushed
from
the
eye.
It
appears
that
scores
were
maximal
at
1
hour
and
24
hours
after
treatment
then
decreased
over
the
rest
of
the
study.

RESULTS
Corrosive:
no
Irritation
score:
Only
total
scores
provided.
1
hr:
8.00;
24
hr:
9.33;
48
hr:
3.33;
72
hr:
zero
Tool
used
to
assess
score:
Not
specified
Description
of
lesions:
none
Remarks:
Inadequate
description
and
discussion
in
report.
Scores
appear
reduced
in
all
rabbits
over
time.
Decreases
were
noted
at
48hrs
and
at
72hrs
values
were
zero.
ENV/
JM/
RD(
2002)
17/
FINAL
278
CONCLUSIONS
Only
conclusion
provided
in
study
is
that
test
substance
is
irritating
to
eyes.
Inadequate
information
is
presented
in
report
to
evaluate
quality
of
study
and
validity
of
conclusion.

REFERENCE
J.
A.
Biesemeier
and
D.
L.
Harris.
1974.
Eye
and
Skin
Iritation
Report
on
Sample
T­
1117.
Project
No.
4102871,
WARF
Institute
Inc.
ENV/
JM/
RD(
2002)
17/
FINAL
279
GENETIC
TOXICITY
STUDIES
Title:
MUTAGENICITY
EVALUATION
OF
T­
2O14
CoC
IN
THE
AMES
SALMONELLA/
MICROSOME
PLATE
TEST,
1978
TEST
SUBSTANCE
Identity:
Potassium
perfluorooctylsulfonate,
CAS
#
2795­
39­
3
Remarks:
T­
2014
CoC,
FC­
95,
Purity
not
specified.

METHOD
Method/
Guideline
followed:
Ames,
1975
Test
type:
Reverse
mutation
Test
system:
Bacteria;
Yeast
GLP:
N
Year
study
performed:
1977
Species/
Strain/
cell­
type/
cell
line:
Salmonella
typhimurium
TA100,
TA1535,
TA1537,
TA1538,
TA09,
Saccharomyces
cerevisiae
D4
Metabolic
activation:
0.1
±
.05
ml
S9
homogenate
of
Aroclor
1254
induced
Sprague
Dawley
rat
liver
Concentrations
tested:
0.01
µ
g/
plate,
1.0
µ
g/
plate,
10.0
µ
g/
plate,
100
µ
g/
plate,
500
µ
g/
plate
nonactivated;
0.1
µ
g/
plate,
1.0
µ
g/
plate,
10.0
µ
g/
plate,
100
µ
g/
plate,
500
µ
g/
plate
activated
Statistical
methods
used:
None
Remarks:
There
were
no
significant
protocol
variations.
(
1)
For
the
time
when
the
test
was
done
(
1977)
a
single
plate
per
concentration
was
routine;
(
2)
the
negative
control
was
the
solvent
DMSO;
the
positive
controls
were
chosen
according
to
strain
being
tested
and
activation
condition
and
included
ethyl
methanesulfonate,
quinoline
hydroxide,
nitroflourene
,
2­
anthramine;
and
2­
dimethylnitrosamine
(
3)
a
limited
repeat
study
was
done
with
strain
TA100
both
with
and
without
activation
because
the
testing
laboratory
believed
that
there
was
some
evidence
of
mutagenicity
with
this
strain.
The
doses
tested
without
activation
were
100
µ
g/
plate,
500
µ
g/
plate,
and
1000
µ
g/
plate
and
500
µ
g/
plate,
1000
µ
g/
plate,
and
2000
µ
g/
plate
with
activation.
However,
a
review
of
the
data
shows
that
was
originally
thought
to
be
mutagenicity
was
within
the
normal
variation
of
the
assay.
The
repeat
was
inadequate
because
the
doses
tested
were
too
high
and
too
toxic
to
shed
any
light
on
possible
mutagenic
activity;
there
were
no
signs
of
mutagenicity
in
any
of
the
other
strains
tested.
The
test
with
Saccharomyces
was
also
negative.
(
4)
criteria
to
evaluate
results
were
as
follows:
dose­
response
over
3
concentrations
with
lowest
increase
equal
to
3X
the
solvent
control
for
TA1535,
TA1537
and
TA1538.
Dose­
response
over
3
concentrations
with
lowest
increase
equal
to
3X
background
for
TA100
and
2x­
3X
background
for
TA98
and
D4.
ENV/
JM/
RD(
2002)
17/
FINAL
280
RESULTS
Overall
results:
positive,
negative,
ambiguous:
Negative
Genotoxic
effects
(
unconfirmed,
dose­
response,
equivocal
 
with/
without
activation):
Negative
with
and
without
activation
Cytotoxic
concentration:
1000
µ
g/
ml
both
with
and
without
activation.

Statistical
results:
No
statistics
performed.

Remarks:
None
CONCLUSIONS
Author's
conclusions
are
accurate
but
for
the
wrong
reasons.
The
test
chemical
is
negative
not
because
it
was
negative
on
repeat
testing
but
because
what
was
taken
as
mutagenicity
in
the
first
test
was
within
normal
variation
of
the
assay.

REFERENCE
Litton
Bionetics,
Inc.
Kensington,
Maryland
20795
1978.
Mutagenicity
Evaluation
of
T­
2014
CoC
in
the
Ames
Salmonella/
Microsome
Plate
Test.
Final
Report.
Submitted
to:
3M
Company,
Saint
Paul,
Minnesota
55101
ENV/
JM/
RD(
2002)
17/
FINAL
281
Title:
SALMONELLA
 
ESCHERICHIA
COLI/
MAMMALIAN­
MICROSOME
REVERSE
MUTATION
ASSAY
WITH
PFOS,
1999
TEST
SUBSTANCE
Identity:
Potassium
perfluorooctylsulfonate,
CAS
#
2795­
39­
3
Remarks:
FC­
95,
T­
6295,
purity
not
specified.

METHOD
Method/
Guideline
followed:
Ames
et
al.,
1975;
Green
and
Muriel,
1976;
Maron
and
Ames,
1983
Test
type:
Reverse
mutation
Test
system:
Bacterial
GLP:
Y
Year
study
performed:
1999
Species/
Strain/
cell­
type/
cell
line:
Salmonella
typhimurium
TA1535,
TA100,
TA98,
TA1537
Escherichia
coli
WP2uvrA
Metabolic
activation:
0.1
ml
S9
liver
homogenate
from
Aroclor
1254
induced
Sprague­
Dawley
rats
Concentrations
tested:
S.
typhimurium:
33.3
µ
g/
plate,
100
µ
g/
plate,
333
µ
g/
plate,
1,000
µ
g/
plate,
3,330
µ
g/
plate,
and
5,000
µ
g/
plate
µ
g/
plate
with
activation
and
0.333
µ
g/
plate,
1.00
µ
g/
plate,
3.33
µ
g/
plate,
10.0
µ
g/
plate,
33.3
µ
g/
plate
plate,
100
µ
g/
plate,
3333
µ
g/
plate,
1,000
µ
g/
plate
and
5,000
µ
g/
plate
without
activation.
E.
coli:
33.3
µ
g/
plate,
100
µ
g/
plate,
3333
µ
g/
plate,
1,000
µ
g/
plate,
3,330
µ
g/
plate,
and
5,000
µ
g/
plate
both
with
and
without
activation.

Statistical
methods
used:
None
Remarks:
There
were
no
significant
protocol
deviations.
(
1)
There
were
3
plates
per
test
concentration
and
control;
the
positive
controls
were
strain
and
activation
condition
specific
and
included
benzo[
a]
pyrene,
2­
nitrofluorene,
2­
aminoanthracene,
sodium
azide,
ICR­
191
and
4­
nitroquinoline­
N­
oxide.
The
vehicle
control
was
DMSO;
(
2)
the
solvent
was
DMSO;
(
3)
the
assay
was
not
repeated.
(
4)
For
the
test
article
to
be
considered
positive
in
strains
TA98,
TA100
and
WP2uvrA,
there
had
to
be
at
least
a
2­
fold
increase
in
the
mean
revertants
per
plate
over
that
of
the
appropriate
vehicle
control.
The
increase
had
to
be
accompanied
by
a
dose
response
to
increasing
concentrations
of
the
test
article.
For
strains
TA1535
and
TA1537
there
had
be
at
least
a
3­
fold
increase
in
the
mean
revertants
per
plate
over
that
of
the
appropriate
vehicle
control.
The
increase
had
to
be
accompanied
by
a
dose
response
to
increasing
concentrations
of
the
test
article.

RESULTS
Overall
results:
positive,
negative,
ambiguous:
Negative
Genotoxic
effects
(
unconfirmed,
dose­
response,
equivocal
 
with/
without
activation):
PFOS
was
not
ENV/
JM/
RD(
2002)
17/
FINAL
282
genotoxic
when
tested
either
with
or
without
metabolic
activation.

Cytotoxic
concentration:
Cytotoxicity
was
noted
at
5000
µ
g/
plate
without
metabolic
activation.
This
cytotoxicity
was
evidenced
by
a
slight
reduction
in
the
bacterial
lawn.

Statistical
results:
Results
were
not
evaluated
statistically.

Remarks:
There
were
no
test­
specific
confounding
factors.
Mutation
frequencies
were
within
the
range
of
the
vehicle
controls.

CONCLUSIONS
Author's
conclusions
are
that
PFOS
is
negative
in
this
assay.
This
is
accurate.

REFERENCE
Mecchi,
M.
S.
1999.
Salmonella
 
Escherichia
Coli/
Mammalian­
Microsome
Reverse
Mutation
Assay
with
PFOS.
Covance
Laboratories
Inc.
(
Covance)
Vienna,
Virginia
22182
Final
Report
Covance
Study
No.:
20784­
0­
409.
Submitted
to:
3M
Corporate
Toxicology
St.
Paul,
Minnesota
55144­
1000
ENV/
JM/
RD(
2002)
17/
FINAL
283
Title:
CHROMOSOMAL
ABERRATIONS
IN
HUMAN
WHOLE
BLOOD
LYMPHOCYTES
WITH
PFOS,
1999
TEST
SUBSTANCE
Identity:
Potassium
perfluorooctylsulfonate,
CAS
#
2795­
39­
3
Remarks:
FC­
95,
purity
not
specified.

METHOD
Method/
Guideline
followed:
Galloway,
1994
Test
type:
In
vitro
cytogenetics
Test
system:
Human
cells
in
culture
GLP:
Y
Year
study
performed:
1999
Species/
Strain/
cell­
type/
cell
line:
Human
lymphocytes
Metabolic
activation:
Aroclor
1254
induced
rat
liver
S9
homogenate,
15.0
µ
L/
ml,
plus
NADP
and
isocitric
acid.

Concentrations
tested:
12.5
µ
g/
ml,
24.9
µ
g/
ml,
49.7
µ
g/
ml,
99.3
µ
g/
ml,
149
µ
g/
ml,
199
µ
g/
ml,
249
µ
g/
ml,
299
µ
g/
ml,
349
µ
g/
ml,
449
µ
g/
ml,
599
µ
g/
ml
without
activation.

12.5
µ
g/
ml,
24.9
µ
g/
ml,
49.7
µ
g/
ml,
99.3
µ
g/
ml,
149
µ
g/
ml,
199
µ
g/
ml,
249
µ
g/
ml,
349
µ
g/
ml,
449
µ
g/
ml
with
activation
Statistical
methods
used:
Cochran­
Armitage
test
for
linear
trend;
Fisher's
Exact
Test
Remarks:
There
were
no
significant
protocol
deviations.
(
1)
Each
concentration
was
tested
in
replicate;
each
replicate
was
considered
an
independent
unit.
The
negative
control
for
the
nonactivation
assay
was
DMSO
at
10
µ
l/
ml,
which
was
the
highest
concentration
used
in
the
test
cultures;
in
the
activation
assay
it
was
DMSO
plus
the
S9
mix;
the
positive
control
was
mitomycin
C
for
the
nonactivation
assay
and
cyclophosphamide
for
the
activation
assay.
Three
concentrations
of
each
positive
control
were
tested.
Cultures
were
exposed
to
chemical
for
3
hours
and
harvested
22
hours
later.
One
hundred
metaphases
from
each
replicate
of
the
useable
treatment
cultures
and
the
solvent
and
one
dose
of
the
positive
control
were
used;
mitotic
index
was
evaluated
by
analysing
the
number
of
mitotic
cells
in
at
least
1000
cells
per
culture;
(
2)
the
solvent
for
the
chemical
was
DMSO;
(
3)
there
was
no
follow
up
study
done
although
in
a
study
such
as
this
where
there
are
negative
results
after
3
hours
incubation
with
a
22
hour
harvest
time
a
second
study
with
a
continuous
exposure
of
22
hours
for
the
nonactivated
portion
of
the
assay
is
recommended.
(
4)
The
test
article
would
have
been
considered
positive
if
there
had
been
a
significant
increase
(
p<
0.01)
in
the
number
of
cells
with
chromosomal
aberrations
at
one
or
more
concentrations.
The
test
article
was
considered
negative
because
there
was
no
significant
increase
observed
in
the
number
of
cells
with
chromosomal
aberrations
at
any
concentration
tested.
ENV/
JM/
RD(
2002)
17/
FINAL
284
RESULTS
Overall
results:
positive,
negative,
ambiguous:
Negative
Genotoxic
effects
(
unconfirmed,
dose­
response,
equivocal
 
with/
without
activation):
Negative
both
with
and
without
activation.

Cytotoxic
concentration:
299
µ
g/
ml
without
metabolic
activation
and
199
µ
g/
ml
with
activation
were
the
first
cytotoxic
concentrations
tested
as
evidenced
by
a
reduction
in
mitotic
index.

Statistical
results:
Negative
Remarks:
Mitotic
index
was
reduced
38%,
8%
15%,
15%,
12%,
19%,
24%,
69%
and
92%
in
cultures
treated
with
12.5
µ
g/
ml,
24.9
µ
g/
ml,
49.7
µ
g/
ml,
99.3
µ
g/
ml,
149
µ
g/
ml,
249
µ
g/
ml,
299
µ
g/
ml,
149
µ
g/
ml
and
449
µ
g/
ml
without
activation.
Aberrations
were
analysed
from
cultures
treated
199
µ
g/
ml,
249
µ
g/
ml,
299
µ
g/
ml,
and
349
µ
g/
ml.

With
metabolic
activation,
mitotic
index
was
reduced
by
12%,
41%,
71%,
and
53%
in
cultures
treated
with
49.7
µ
g/
ml,
199
µ
g/
ml,
249
µ
g/
ml,
and
299
µ
g/
ml.
Aberrations
were
analysed
from
cultures
treated
with
99.3
µ
g/
ml,
149
µ
g/
ml,
199
µ
g/
ml,
and
299
µ
g/
ml.
Only
27
and
4
metaphases
were
available
for
analysis
from
cultures
treated
with
299
µ
g/
ml.

CONCLUSIONS
Author's
conclusions
are
that
PFOS
does
not
cause
mutation
in
human
lymphocytes;
this
is
correct
as
stated.

REFERENCE
Murli,
H.
1999.
Chromosomal
Aberrations
in
Human
Whole
Blood
Lymphocytes
with
PFOS.
Covance
Laboratories
Inc.
(
Covance)
Final
Report.
Covance
Study
No.:
20784­
0­
449.
Submitted
to:
3M
Corporate
Toxicology,
St.
Paul
Minnesota
55144­
1000.
ENV/
JM/
RD(
2002)
17/
FINAL
285
Title:
Unscheduled
DNA
Synthesis
in
Rat
Liver
Primary
Cell
Cultures
with
PFOS,
1999
TEST
SUBSTANCE
Identity:
Potassium
perfluorooctylsulfonate,
CAS
#
2795­
39­
3
Remarks:
T­
6295,
purity
not
specified.

METHOD
Method/
Guideline
followed:
Williams,
1977;
Williams,
1980;
Butterworth
et
al.,
1987
Test
type:
Unscheduled
DNA
Synthesis
in
Mammalian
Cells
in
Culture
Test
system:
Primary
cells
in
culture
GLP:
Y
Year
study
performed:
1999
Species/
Strain/
cell­
type/
cell
line:
Primary
hepatocytes
from
a
Fischer
344
rat
male
rat.

Metabolic
activation:
None
Concentrations
tested:
15
concentrations
between
0.025
µ
g/
ml
and
4000
µ
g/
ml.
Six,
0.5
µ
g/
ml,
1.0
µ
g/
ml,
2.5
µ
g/
ml,
5
µ
g/
ml,
10.0
µ
g/
ml
and
25.0
µ
g/
ml,
chosen
for
evaluation
based
upon
cytotoxicity.

Statistical
methods
used:
None
Remarks:
There
were
no
significant
protocol
deviations.
(
1)
Triplicate
cultures
on
coverslips
were
incubated
for
19.6­
20.0
hours,
then
the
assay
was
terminated
and
3H­
thymidine
added
to
the
cultures
for
30
minutes
after
which
the
cells
were
fixed,
dried
over
night,
coverslips
were
mounted
on
slides,
dipped
in
emulsion
and
stored
for
6
days
at
2­
8o
C
after
which
the
emulsions
were
developed,
fixed
and
stained.
150
cells
per
dose
were
read
(
50
from
each
coverslip)
and
the
mean
net
nuclear
grain
count
determined.
(
2)
The
solvent
for
the
assay
was
DMSO;
(
3)
there
was
no
follow­
up
repeat
study;
(
4)
the
positive
control
was
2­
AAF;
(
5)
for
a
treatment
to
be
considered
positive,
there
must
be
an
increase
in
the
mean
net
nuclear
grain
count
to
at
least
5
grains
per
nucleus
above
the
concurrent
vehicle
control
value,
and/
or
an
increase
in
the
number
of
nuclei
with
five
or
more
net
grains
such
that
the
percentage
of
these
nuclei
in
test
cultures
is
10%
above
the
percentage
seen
in
the
vehicle
control
cultures.
The
positive
control
satisfied
both
of
these
criteria.

RESULTS
Overall
results:
positive,
negative,
ambiguous:
Negative
Genotoxic
effects
(
unconfirmed,
dose­
response,
equivocal
 
with/
without
activation):
Negative
Cytotoxic
concentration:
Excessive
cytotoxicity
at
and
at
50.0
µ
g/
ml;
weak
cytotoxicity
at
25.0
µ
g/
ml.
Cell
morphology
was
suitable
for
analysis
at
and
below
25.0
µ
g/
ml.
ENV/
JM/
RD(
2002)
17/
FINAL
286
Statistical
results:
The
results
were
not
evaluated
statistically.

Remarks:
There
were
no
test­
specific
confounding
factors.

CONCLUSIONS
The
author
concludes
that
PFOS
is
negative
in
this
assay.
This
is
accurate.

REFERENCE
Cifone
M.
A.
1999.
Unscheduled
DNA
Synthesis
in
Rat
Liver
Primary
Cell
Cultures
with
PFOS.
Covance
Laboratories
Inc.
Vienna,
VA
22182
Final
Report.
Covance
Study
No.:
20780­
0­
447.
Submitted
to
3M
Corporate
Toxicology
St.
Paul,
MN
55144­
1000
ENV/
JM/
RD(
2002)
17/
FINAL
287
Title:
MUTAGENICITY
TEST
ON
T­
6295
IN
AN
IN
VIVO
MOUSE
MICRONUCLEUS
ASSAY,
1996
TEST
SUBSTANCE
Identity:
Potassium
perfluorooctylsulfonate,
CAS
#
2795­
39­
3
Remarks:
T­
6295,
FC­
95,
purity
not
specified.

METHOD
Method/
Guideline
followed:
Heddle,
1983
Test
type:
Micronucleus
GLP:
Y
Year
study
performed:
1996
Species/
Strain:
Mouse;
Crl:
CD­
1
®
(
ICR)
BR
Sex:
Males
&
Females
No.
animals/
sex/
dose:
5/
sex/
dose
Vehicle
(
if
used):
Deionized
water
Route
of
administration:
Oral
Doses:
237.5
mg/
kg,
450
mg/
kg,
950
mg/
kg
Frequency
of
treatment:
Single
dose
Statistical
methods
used:
Analysis
of
variance;
Dunnet's
t­
test
Remarks:
There
were
no
significant
protocol
deviations.
(
1)
Animals
were
9
weeks
and
1
day
old
at
start
of
dosing
males;
weight
range
for
the
males
was
29.9
 
37.0
g;
for
females
it
was
23.1­
29.2
g;
(
2)
the
vehicle
was
deionized
water;
(
3)
the
test
lasted
72
hours;
(
4)
the
test
material
was
administered
as
a
single
oral
dose;
(
5)
all
treatment
groups
were
sampled
at
24,
48
and
72
hours;
(
6)
the
vehicle
control
was
H2O;
the
positive
control
was
80
mg/
kg
cyclophosphamide
dissolved
in
water
and
administered
by
gavage.
Controls
were
sampled
at
24
hours
only.
Control
groups
consisted
of
5
males
and
5
females
each.
(
7)
No
clinical
examinations
were
made.
(
8)
No
necropsies
or
other
gross
examinations
were
made
on
these
animals.
(
9)
Micronuclei
were
evaluated
in
the
bone
marrow
of
treated
animals.
Frequency
of
PCEs
vs.
NCEs
was
determined
by
scoring
the
number
of
PCEs
and
NCEs
in
the
optic
fields
while
scoring
the
first
1000
erythrocytes.
A
positive
was
judged
by
an
increase
in
micronucleated
polychromatic
erythrocytes
over
levels
observed
in
the
vehicle
controls
in
either
sex
or
at
any
harvest
time.
Bone
marrow
toxicity
was
judged
by
a
significant
reduction
in
PCE/
NCE
ratios
in
either
sex
at
any
harvest
time.
(
10)
The
M.
T.
D.
was
chosen
on
the
basis
of
2
preliminary
dose
selection
assays
both
of
which
showed
significant
toxicity
at
the
highest
dose
tested.
ENV/
JM/
RD(
2002)
17/
FINAL
288
RESULTS
Effect
on
mitotic
index
or
PCE/
NCE
ratio
by
dose
level
and
sex:
PCE:
NCE
Ratio
237.5
mg/
kg
24
hours:
males
0.57
±
0.11;
females
0.52
±
0.10
48
hours:
males
0.48
±
0.04;
females
0.80
±
0.10
72
hours:
males
0.39
±
0.11;
females
0.42
±
0.14
450
mg/
kg
24
hours:
males
0.75
±
0.11;
females
0.59
±
0.08
48
hours:
males
0.71
±
0.05;
females
0.37
±
0.07
72
hours:
males
0.29
±
0.06;
females
0.40
±
0.12
950
mg/
kg
24
hours:
males
0.56
±
0.13;
females
0.59
±
0.08
48
hours:
males
0.54
±
0.08;
females
0.44
±
0.11
72
hours:
males
0.17
±
0.05;
females
0.17
±
0.05
Genotoxic
effects
(
unconfirmed,
dose­
response,
equivocal):
Negative
Statistical
results:
The
PCE:
NCE
ratio
was
reduced
in
237.5
mg/
kg
males
at
48
and
72
hours;
in
450
mg/
kg
males
at
72
hours
and
in
450
mg/
kg
females
at
48
hours
and
in
950
mg/
kg
males
at
48
and
72
hours
and
in
950
mg/
kg
females
at
72
hours.
There
was
no
statistically
significant
increase
in
the
number
of
micronucleated
PCEs
over
the
controls
in
any
treatment
group.
The
positive
control
induced
a
significant
increase
in
the
number
of
mPCE
in
both
males
and
females
and
reduced
the
PCE:
NCE
ratio
in
females
only
at
24
hours.

Remarks:
(
1)
Animals
were
examined
approximately
1­
2
hours
before
sampling
for
signs
of
toxicity
and
mortality.
Animals
in
the
237.5
mg/
kg
group
remained
healthy
throughout
the
treatment
period.
(
2)
Both
males
and
females
in
the
950
mg/
kg
dose
group
began
dying
about
22
hours
after
treatment.
Also
at
22
hours
2
males
in
the
950
mg/
kg
dose
group
went
into
convulsions
when
their
cage
was
opened
but
recovered
in
a
few
minutes.
At
about
46
hours
after
treatment
1
female
from
the
450
mg/
kg
dose
group
and
more
males
and
females
from
the
950
mg/
kg
dose
group
were
found
dead
and
at
about
71
hours
after
treatment,
one
male
from
the
950
mg/
kg
dose
group
was
found
dead.
All
surviving
animals
appeared
normal
at
that
point.
(
3)
No
other
clinical
signs
were
noted
or
reported.
(
4)
Body
weight
changes
were
not
reported.
(
5)
Food
and
water
consumption
were
not
reported.
(
6)
There
was
no
increase
in
the
percent
of
micronucleated
PCEs
at
any
dose
level
tested
or
at
any
time
period
sampled.

CONCLUSIONS
The
author
concludes
that
PFOS
is
negative
in
the
mouse
bone
marrow
micronucleus
assay.
This
is
an
accurate
assessment.

REFERENCE
Murli,
H.
1996.
Mutagenicity
Test
on
T­
6295
in
an
In
Vivo
Mouse
Micronucleus
Assay.
Corning
Hazelton
Inc.
(
CHV),
Vienna,
Virginia
22182.
Final
Report.
CHV
Study
No.:
17403­
0­
455.
Submitted
to
3M
St.
Paul,
Minnesota
55144­
1000.
ENV/
JM/
RD(
2002)
17/
FINAL
289
Title:
IN
VITRO
MICROBIOLOGICAL
MUTAGENICITY
ASSAYS
OF
3M
COMPANY
COMPOUNDS
T­
2247
CoC
AND
T­
2248
CoC,
1978
TEST
SUBSTANCE
Identity:
T­
2247
CoC;
L­
4299,
a
50%
by
weight
solution
of
the
diethanolammonium
salt
of
perfluorooctanesulfonate
in
water
T­
2248
CoC;
22.5%
of
a
reaction
product
of
ethyl
and
methyl
methacrylates
and
22.5%
of
the
pyridinium
chloride
salt
of
an:
N­
methylperfluorooctanesulfonamidoethanol­
based
glutaryl
amide.

Remarks:
T­
2247
CoC,
T­
2248
CoC,
purity
not
specified
METHOD
Method/
Guideline
followed:
Ames
et
al.,
1975;
Zimmermann
and
Schwaier,
1967;
Brusick
and
Mayer,
1973
Test
type:
Reverse
Mutation;
Recombination
Test
system:
Salmonella
typhimurium;
Saccharomyces
cerevisiae
GLP:
N
Year
study
performed:
1978
Species/
Strain/
cell­
type/
cell
line:
Salmonella
typhimurium
TA1535,
TA1537,
TA1538,
TA98,
TA100;
Saccharomyces
cerevisiae
D3
Metabolic
activation:
0.5
ml
of
10%
S9
liver
homogenate
from
Aroclor
1254
induced
rats.

Concentrations
tested:
Plate
incorporation
assay:
10
µ
g/
plate,
50
µ
g/
plate,
100
µ
g/
plate,
500
µ
g/
plate,
1000
µ
g/
plate,
5000
µ
g/
plate
Dessicator
method:
0.1
ml/
dessicator,
0.5
ml/
dessicator,
1.0
ml/
dessicator,
5.0
ml/
dessicator
Yeast
recombination:
0.1%,
0.5%,
1.0%,
5.0%
Yeast
repeat
assay
at
1.0%,
2.0%,
4.0%,
5.0%

Statistical
methods
used:
None
Remarks:
There
were
no
significant
protocol
deviations.
(
1).
The
plate
incorporation
assay
and
the
S.
cerevisiae
assay
were
performed
with
both
chemicals
and
with
one
plate
per
test
concentration;
the
dessicator
assay
was
performed
with
T­
2247
CoC
using
two
plates
per
concentration
but
used
only
strains
TA
98
and
TA100
for
the
test.
However,
given
the
complexity
of
the
dessicator
assay
and
the
limitations
involved
in
setting
it
up,
this
is
acceptable;
(
2)
the
positive
controls
were
chosen
according
to
the
strain
and
activation
conditions
and
included
sodium
azide,
9­
aminoacridine,
2­
nitrofluorene
and
2­
anthramine
for
the
plate
incorporation
assay;
1,1­
dichloroethylene
for
the
desiccator
assay
with
T­
2247
CoC
and
1,2,3,4­
diepoxybutane
for
the
S.
cerevisiae
assay.
The
negative
control
group
for
all
assays
was
water.
(
3)
The
plate
incorporation
assay
with
both
agents
and
the
yeast
assay
with
T­
2248
were
repeated;
the
desiccator
assay
was
run
only
once.
(
4)
For
the
desiccator
assay,
plates
were
prepared
as
for
the
standard
assay
but
no
test
chemical
was
added
to
the
agar.
The
strains
tested
were
S.
typhimurium
TA98
and
TA
100.
The
test
was
performed
both
with
and
without
metabolic
activation.
Plates
without
lids
were
placed
side
by
side
in
ENV/
JM/
RD(
2002)
17/
FINAL
290
a
perforated
shelf
in
a
9­
liter
desiccator.
A
known
volume
of
T­
2247
was
added
to
a
glass
Petri
dish
that
was
placed
in
the
center
of
and
attached
to
the
bottom
of
the
shelf.
In
decreasing
order,
5.0
ml,
1.0
ml,
0.5
ml
and
0.1
ml
of
test
chemical
were
added
to
the
desiccator.
The
negative
control
chemical
was
water;
the
positive
control
chemical
was
1,1­
dichloroethylene.
Both
were
treated
in
the
same
manner
as
T­
2247.
The
desiccator
was
sealed
and
placed
on
a
magnetic
stirrer
plate
in
a
room
maintained
at
37o
C.
A
magnetic
stirrer
with
vanes
was
placed
in
the
base
of
each
desiccator
to
ensure
adequate
dispersion
of
the
chemical.
Plates
were
incubated
for
8
hours,
removed
from
the
desiccators,
their
lids
replaced
and
they
were
incubated
at
37o
C
for
an
additional
42
hours
before
revertants
were
counted.

RESULTS
Overall
results:
positive,
negative,
ambiguous:
All
tests
were
negative.

Genotoxic
effects
(
unconfirmed,
dose­
response,
equivocal
 
with/
without
activation):
Negative
both
with
and
without
activation.

Cytotoxic
concentration:
T­
2247
was
not
cytotoxic.
In
the
plate
incorporation
assay,
T­
2248
was
toxic
to
strain
TA1538
at
1000
µ
g/
plate
and
to
all
other
strains
at
5000
µ
g/
plate
when
tested
without
activation.
It
was
toxic
at
1000
µ
g/
plate
to
strain
TA1537
and
at
5000
µ
g/
plate
for
all
other
strains
when
tested
with
metabolic
activation.
T­
2248
was
slightly
toxic
to
S.
cerevisiae
D3
at
5%
concentration
without
metabolic
activation.

Statistical
results:
No
statistical
results
were
determined.

Remarks:
In
the
first
assay
with
T­
2248
and
S.
cerevisiae
D3
without
metabolic
activation
there
seemed
to
be
some
slight
indication
of
mutagenicity
at
the
highest
concentration
tested,
5%.
The
assay
was
repeated
at
1%,
2%,
4%,
and
5%
concentrations
with
and
without
activation.
There
was
no
indication
of
a
mutagenic
dose
response
and
the
testing
laboratory
concluded
that
T­
2248
did
not
cause
recombination
in
S.
cerevisiae
D3.
There
were
no
test­
specific
confounding
factors
in
any
aspect
of
the
test.

CONCLUSIONS
The
testing
laboratory
concluded
that
T­
2247
and
T­
2248
were
nonmutagenic
for
S.
typhimurium
TA1535,
TA100,
TA1537,
TA1538,
and
TA98
when
tested
in
a
plate
incorporation
assay
with
and
without
metabolic
activation;
that
T2247
did
not
induce
mutation
in
S.
typhimurium
TA98
and
TA100
when
tested
in
a
dessicator
assay
for
volatile
chemical
and
that
neither
chemical
induced
recombination
in
S.
cerevisiae
D3.
These
conclusions
are
accurate.

REFERENCE
Simmon,
V.
F.
1978.
IN
VITRO
MICROBIOLOGICAL
MUTAGENICITY
ASSAYS
OF
3M
COMPANY
COMPOUNDS
T­
2247
CoC
AND
T­
2248
CoC.
SRI
International,
Final
Report.
Prepared
for
3M
Company,
St.
Paul,
Minnesota
55101.
ENV/
JM/
RD(
2002)
17/
FINAL
291
REPEAT
DOSE
STUDIES
Title:
First
ninety­
day
rhesus
monkey
toxicity
study,
1979
TEST
SUBSTANCE
Identity:
Potassium
perfluorooctylsulfonate,
CAS
#
2795­
39­
3
Remarks:
FC­
95,
purity
not
specified.

METHOD
Method/
guideline
followed:
None
Study
duration:
90
days
GLP
(
Y/
N):
No
Year
study
performed:
1978
Species/
strain:
Rhesus
monkey
Sex:
Males
and
females
Number
of
animals
per
dose
group:
2/
sex/
group
Route
of
administration:
Gavage
Doses
tested
and
frequency:
0,
10,
30,
100,
300
mg/
kg/
day
Post­
observation
period:
None
Statistical
methods
used:
None
Remarks:
Distilled
water
was
used
for
the
vehicle
control.
The
males
weighed
3.05­
3.80
kg
at
study
initiation
and
the
females
weighed
2.75­
4.10
kg.
The
monkeys
were
observed
daily
for
general
clinical
signs
and
body
weights
were
recorded
weekly.
Hematological
and
clinical
chemistry
analyses
and
urinalysis
were
conducted
at
the
beginning
of
the
study.
The
study
was
terminated
after
20
days
due
to
the
death
of
the
monkeys.
At
necropsy
the
heart,
liver,
adrenals,
spleen,
pituitary,
kidneys,
testes/
ovaries
and
brain
were
weighed.
The
thyroid/
parathyroid
were
weighed
after
fixation.
Tissues
were
preserved
in
buffered
neutral
10%
formalin;
the
eyes
were
preserved
in
Russell's
fixative.
The
following
organs
from
control
and
all
treated
groups
were
examined
microscopically:
adrenals,
aorta,
brain,
esophagus,
eyes,
gallbladder,
heart
(
with
coronary
vessels),
duodenum,
ileum,
jejunum,
cecum,
colon,
rectum,
kidneys,
liver,
lung,
skin,
mesenteric
lymph
node,
retropharyngeal
lymph
node,
mammary
gland,
nerve
(
with
muscle),
spleen,
pancreas,
prostate/
uterus,
bone/
bone
marrow
(
rib
junction),
salivary
gland,
lumbar
spinal
cord,
pituitary,
stomach,
testes/
ovaries,
thyroid,
parathyroid,
thymus,
trachea,
tonsil,
tongue,
urinary
bladder
and
vagina.
ENV/
JM/
RD(
2002)
17/
FINAL
292
RESULTS
NOAEL
(
dose
and
effect):
None
LOAEL
(
dose
and
effect):
None
Toxic
response/
effects
by
dose
level:
All
of
the
monkeys
in
the
treated
groups
died.

Statistical
results:
None
Remarks:
The
monkeys
in
the
300
mg/
kg/
day
group
died
between
days
2­
4,
the
monkeys
in
the
100
mg/
kg/
day
group
died
between
days
3­
5,
the
monkeys
in
the
30
mg/
kg/
day
group
died
between
days
7­
10,
and
the
monkeys
in
the
10
mg/
kg/
day
group
died
between
days
11­
20
of
treatment.
The
monkeys
from
all
the
groups
showed
similar
signs
of
toxicity
including
decreased
activity,
emesis
with
some
diarrhea,
body
stiffening,
general
body
trembling,
twitching,
weakness,
convulsions
and
prostration.
At
necropsy,
several
of
the
monkeys
in
the
100
and
300
mg/
kg/
day
groups
had
a
yellowish­
brown
discoloration
of
the
liver;
histologic
examination
showed
no
microscopic
lesions.
Congestion,
hemorrhage
and
lipid
depletion
of
the
adrenal
cortex
was
noted
in
all
treated
groups.
No
other
lesions
were
noted.

CONCLUSIONS
Remarks:
Authors
conclusions
stated
above
in
results.
Reviewer
agrees.

REFERENCE
Goldenthal,
E.
I.,
D.
C.
Jessup,
R.
G.
Geil
and
J.
S.
Mehring.
1979.
Ninety­
day
subacute
rhesus
monkey
toxicity
study.
Study
No.
137­
087,
International
Research
and
Development
Corporation,
Mattawan,
MI.
ENV/
JM/
RD(
2002)
17/
FINAL
293
Title:
Second
ninety­
day
rhesus
monkey
toxicity
study,
1978
TEST
SUBSTANCE
Identity:
Potassium
perfluorooctylsulfonate,
CAS
#
2795­
39­
3
Remarks:
FC­
95,
purity
not
specified.

METHOD
Method/
guideline
followed:
None
Study
duration:
90
days
GLP
(
Y/
N):
No
Year
study
performed:
1978
Species/
strain:
Rhesus
monkey
Sex:
Males
and
females
Number
of
animals
per
dose
group:
2/
sex/
group
Route
of
administration:
Gavage
Doses
tested
and
frequency:
0,
0.5,
1.5,
4.5
mg/
kg/
day
Post­
observation
period:
None
Statistical
methods
used:
Body
wts,
hematological,
biochemical
and
urinalysis
and
organ
wts
were
compared
by
analysis
of
variance
(
one­
way
classification),
Bartlett's
test
and
the
appropriate
t­
test
using
Dunnett's
multiple
comparison
tables
to
judge
significance
of
differences.

Remarks:
Distilled
water
was
used
for
the
vehicle
control.
The
males
weighed
2.55­
3.55
kg
at
study
initiation
and
the
females
weighed
2.7­
3.75
kg.
The
monkeys
were
observed
daily
for
general
clinical
signs
and
body
weights
were
recorded
weekly.
Hematological
and
clinical
chemistry
analyses
and
urinalysis
were
conducted
at
the
beginning
of
the
study
and
after
30
and
90
days
of
treatment.
At
necropsy
the
heart,
liver,
adrenals,
spleen,
pituitary,
kidneys,
testes/
ovaries
and
brain
were
weighed.
The
thyroid/
parathyroid
were
weighed
after
fixation.
Tissues
were
preserved
in
buffered
neutral
10%
formalin;
the
eyes
were
preserved
in
Russell's
fixative.
The
following
organs
from
control
and
all
treated
groups
were
examined
microscopically:
adrenals,
aorta,
brain,
esophagus,
eyes,
gallbladder,
heart
(
with
coronary
vessels),
duodenum,
ileum,
jejunum,
cecum,
colon,
rectum,
kidneys,
liver,
lung,
skin,
mesenteric
lymph
node,
retropharyngeal
lymph
node,
mammary
gland,
nerve
(
with
muscle),
spleen,
pancreas,
prostate/
uterus,
bone/
bone
marrow
(
rib
junction),
salivary
gland,
lumbar
spinal
cord,
pituitary,
stomach,
testes/
ovaries,
thyroid,
parathyroid,
thymus,
trachea,
tonsil,
tongue,
urinary
bladder
and
vagina.

RESULTS
NOAEL
(
dose
and
effect):
None
ENV/
JM/
RD(
2002)
17/
FINAL
294
LOAEL
(
dose
and
effect):
0.5
mg/
kg/
day:
Soft
stools,
diarrhea,
anorexia,
emesis,
occasional
decreases
in
activity;
slight
reduction
in
serum
alkaline
phosphatase.

Toxic
response/
effects
by
dose
level:
4.5
mg/
kg/
day
­
4/
4
monkeys
died
between
weeks
5­
7,
clinical
signs
(
anorexia,
emesis,
black
stool,
dehydration),
significant
reduction
in
serum
cholesterol,
marked
diffuse
lipid
depletion
in
the
adrenals,
moderate
diffuse
atrophy
of
pancreatic
acinar
cells,
moderate
diffuse
atrophy
of
serous
alveolar
cells.
1.5
mg/
gk/
day
 
clinical
signs
(
soft
stools,
diarrhea),
reduced
body
weight,
reduced
serum
alkaline
phosphatase
activity
and
serum
potassium
(
females),
reduced
serum
cholesterol
(
1/
2
females),
reduced
inorganic
phosphate
(
1/
2
females).
0.5
mg/
kg/
day
 
clinical
signs,
(
soft
stools,
diarrhea),
soft
stools,
diarrhea,
anorexia,
emesis,
occasional
decreases
in
activity;
slight
reduction
in
serum
alkaline
phosphatase.

Statistical
results:
The
statistical
results
are
presented,
but
should
be
viewed
with
caution
due
to
the
small
number
of
animals.
4.5
mg/
kg/
day
­
significant
reduction
in
serum
cholesterol
1.5
mg/
kg/
day
 
significant
reduction
in
serum
alkaline
phosphatase
activity
and
serum
potassium
(
females)

Remarks:
All
monkeys
in
the
4.5
mg/
kg/
day
group
died
or
were
sacrificed
in
extremis
between
week
5
and
7
of
the
study.
Beginning
on
the
first
or
second
day
of
the
study,
these
monkeys
exhibited
signs
of
gastrointestinal
tract
toxicity
including
anorexia,
emesis,
black
stool
and
dehydration.
All
of
the
monkeys
had
decreased
activity
and
just
prior
to
death
showed
marked
to
severe
rigidity,
convulsions,
generalized
body
trembling
and
prostration.
The
mean
body
weight
decreased
from
3.44
kg
at
the
beginning
of
the
study
to
2.7
kg
at
week
5.
After
30
days
of
treatment,
there
was
a
significant
reduction
in
serum
cholesterol
and
a
50%
reduction
in
serum
alkaline
phosphatase
activity.
At
necropsy,
mean
organ
weights
were
comparable
among
the
control
and
treated
monkeys.
Histologic
examination
showed
several
treatment
related
lesions.
All
the
male
and
females
had
marked
diffuse
lipid
depletion
in
the
adrenals.
One
male
and
two
females
had
moderate
diffuse
atrophy
of
the
pancreatic
exocrine
cells
with
decreased
cell
size
and
loss
of
zymogen
granules.
Two
males
and
one
female
had
moderate
diffuse
atrophy
of
the
serous
alveolar
cells
characterized
by
decreased
cell
size
and
loss
of
cytoplasmic
granules.

All
monkeys
in
the
1.5
mg/
kg/
day
group
survived
until
the
end
of
the
study.
During
the
first
week
of
the
study,
the
monkeys
had
decreased
activity.
Signs
of
gastrointestinal
tract
toxicity
were
noted
occasionally
during
the
study
and
included
black
stool,
diarrhea,
mucous
in
the
stool
and
bloody
stool;
at
the
end
of
the
study,
anorexia,
dehydration
or
general
body
trembling
were
noted.
Although
statistical
significance
was
not
achieved,
the
mean
body
weight
of
the
males
dropped
from
3.15
kg
at
the
beginning
of
the
study
to
2.93
kg
at
the
end
of
the
study,
and
the
mean
body
weight
of
the
females
dropped
from
3.22
kg
to
2.75
kg.
One
of
the
females
had
very
low
serum
cholesterol
and
another
had
a
reduction
in
inorganic
phosphate.
Necropsy
revealed
no
treatment
related
lesions.

All
monkeys
in
the
0.5
mg/
kg/
day
group
survived
until
the
end
of
the
study.
Signs
of
gastrointestinal
tract
toxicity
were
noted
occasionally
during
the
study
and
included
diarrhea,
soft
stools,
anorexia
and
emesis.
Occasionally,
decreased
activity
was
noted
in
three
of
the
monkeys.
Necropsy
revealed
no
treatment
related
lesions.

CONCLUSIONS
Remarks:
Authors
conclusions
stated
above
in
results.
Reviewer
agrees.
ENV/
JM/
RD(
2002)
17/
FINAL
295
REFERENCE
Goldenthal,
E.
I.,
D.
C.
Jessup,
R.
G.
Geil
and
J.
S.
Mehring.
1978.
Ninety­
day
subacute
rhesus
monkey
toxicity
study.
Study
No.
137­
092,
International
Research
and
Development
Corporation,
Mattawan,
MI.
ENV/
JM/
RD(
2002)
17/
FINAL
296
Title:
Ninety
day
study
in
rats,
1978
TEST
SUBSTANCE
Identity:
Potassium
perfluorooctylsulfonate,
CAS
#
2795­
39­
3
Remarks:
FC­
95,
purity
not
specified.

METHOD
Method/
guideline
followed:
None
Study
duration:
90
days
GLP
(
Y/
N):
No
Year
study
performed:
1978
Species/
strain:
CD
rat
Sex:
Males
and
females
Number
of
animals
per
dose
group:
5/
sex/
group
Route
of
administration:
Diet
Doses
tested
and
frequency:
0,
30,
100,
300,
1000,
3000
ppm
Equivalent
to
0,
2,
6,
18,
60,
200
mg/
kg/
day
Post­
observation
period:
None
Statistical
methods
used:
Body
wts,
hematological,
biochemical
and
urinalysis
and
organ
wts
were
compared
by
analysis
of
variance
(
one­
way
classification),
Bartlett's
test
and
the
appropriate
t­
test
using
Dunnett's
multiple
comparison
tables
to
judge
significance
of
differences.

Remarks:
The
males
weighed
196­
232
g
and
the
females
weighed
165­
206
g
at
study
initiation.
The
animals
were
observed
daily
for
general
clinical
signs
and
body
weights
were
recorded
weekly.
Hematological
and
clinical
chemistry
analyses
and
urinalysis
were
conducted
at
the
beginning
of
the
study
and
after
30
and
90
days
of
treatment.
At
necropsy
the
heart,
liver,
adrenals,
spleen,
pituitary,
kidneys,
testes/
ovaries
and
brain
were
weighed.
The
thyroid/
parathyroid
were
weighed
after
fixation.
Tissues
were
preserved
in
buffered
neutral
10%
formalin;
the
eyes
were
preserved
in
Russell's
fixative.
The
following
organs
from
control
and
all
treated
groups
were
examined
microscopically:
adrenals,
aorta,
brain,
esophagus,
eyes,
gallbladder,
heart
(
with
coronary
vessels),
duodenum,
ileum,
jejunum,
cecum,
colon,
rectum,
kidneys,
liver,
lung,
skin,
mesenteric
lymph
node,
retropharyngeal
lymph
node,
mammary
gland,
nerve
(
with
muscle),
spleen,
pancreas,
prostate/
uterus,
bone/
bone
marrow
(
rib
junction),
salivary
gland,
lumbar
spinal
cord,
pituitary,
stomach,
testes/
ovaries,
thyroid,
parathyroid,
thymus,
trachea,
tonsil,
tongue,
urinary
bladder
and
vagina.
ENV/
JM/
RD(
2002)
17/
FINAL
297
RESULTS
NOAEL
(
dose
and
effect):
None
LOAEL
(
dose
and
effect):
30
ppm
(
2
mg/
kg/
day)
based
on
a
significant
increase
in
relative
and
absolute
liver
weights.

Toxic
response/
effects
by
dose
level:
3000
ppm
 
10/
10
rats
died
between
days
7­
8.
1000
ppm
 
10/
10
rats
died
between
days
8­
14.
300
ppm
 
5/
5
male
rats
died
between
days
13­
25;
5/
5
female
rats
died
between
days
18­
28.
At
300,
1000
and
3000
ppm
 
histologic
lesions
in
the
primary
(
thymus,
bone
marrow)
and
secondary
(
spleen,
mesenteric
lymph
nodes)
lymphoid
organs,
stomach,
intestines,
muscle
and
skin.
100
ppm
 
2/
5
males
and
2/
5
females
died
during
week
5
and
a
third
male
died
during
week
11,
mean
body
weights
were
reduced
by
16.7%
(
males)
and
16.3%
(
females)
at
study
termination,
food
consumption
significantly
reduced,
significant
reduction
in
hematocrit
(
males),
erythrocyte
(
males),
hemoglobin
(
males
&
females),
leukocyte
(
males),
and
reticulocyte
(
females)
counts,
significant
increase
in
absolute
(
females)
and
relative
(
males
&
females)
liver
weight
and
relative
kidney
weight.
At
100,
300,
1000
and
3000
ppm
 
slight
to
marked
focal
necrosis
of
hepatocytes.
30
ppm
 
Significant
reduction
in
food
consumption
(
males),
significant
increase
in
absolute
and
relative
liver
weight
(
females).
At
all
dose
levels
 
very
slight
to
slight
cytoplasmic
hypertrophy
of
hepatocytes
in
the
centrilobular
ro
midzonal
regions,
especially
in
males.

Statistical
results:
100
ppm
 
significant
reduction
in
food
consumption
Remarks:
All
of
the
rats
in
the
300,
1000
and
3000
ppm
groups
died.
Death
occurred
between
days
13­
25
and
days
18­
28
for
the
males
and
females,
respectively,
in
the
300
ppm
group.
At
1000
ppm,
death
occurred
between
days
8­
14,
and
at
3000
ppm,
the
rats
died
between
days
7­
8
of
treatment.
The
rats
in
all
groups
showed
signs
of
toxicity
including
emaciation,
convulsions
following
handling,
hunched
back,
red
material
around
the
eyes,
yellow
material
around
the
anogenital
region,
increased
sensitivity
to
external
stimuli,
reduced
activity
and
moist
red
material
around
the
mouth
or
nose.

Three
males
and
two
females
in
the
100
ppm
group
died
prior
to
scheduled
sacrifice.
Two
of
the
males
and
the
two
females
died
during
week
5
and
the
third
male
died
during
week
11
of
the
study.
At
study
termination,
mean
body
weights
were
reduced
by
16.7%
and
16.3%
in
the
male
and
female
groups,
respectively.
Average
food
consumption
during
the
entire
study
period
(
g/
rat/
day)
was
significantly
reduced
for
males
and
females
at
100
ppm.
After
30
days
of
treatment,
hematologic
values
were
comparable
among
the
control
and
100
ppm
groups.
Clinical
chemistry
analyses
at
one
month
showed
a
significant
increase
in
mean
glucose
in
males,
blood
urea
nitrogen
values
in
males
and
females,
and
creatinine
phosphokinase
and
alkaline
phosphatase
values
for
females.
After
90
days
of
treatment
at
100
ppm,
the
two
surviving
males
had
significantly
reduced
erythrocyte,
hemoglobin,
hematocrit
and
leukocyte
counts;
the
three
surviving
females
had
significantly
reduced
hemoglobin
and
reticulocyte
counts,
as
well
as
slightly
lower
erythrocyte,
hematocrit
and
leukocyte
counts.
Two
of
the
surviving
females
showed
slight
to
moderate
increases
in
plasma
glutamic
oxalacetic
and
pyruvic
transaminase
activities.
Urinalysis
results
were
comparable
among
treated
and
control
groups
at
30
and
90
days.
Relative
liver
weight
was
significantly
increased
in
the
males
and
absolute
and
relative
liver
weights
were
significantly
increased
in
the
females..
Relative
kidney
weights
were
significantly
increased
in
both
sexes.

All
rats
in
the
30
ppm
group
survived
until
the
end
of
the
study.
At
study
termination,
mean
body
weights
were
reduced
by
8.7
and
8%
in
the
males
and
females,
respectively.
Average
food
consumption
during
the
ENV/
JM/
RD(
2002)
17/
FINAL
298
entire
study
period
(
g/
rat/
day)
was
significantly
reduced
for
the
males
at
30
ppm.
Hematologic
values
were
comparable
among
the
control
and
30
ppm
group
at
30
and
90
days.
One
female
showed
a
slightly
elevated
glucose
level
and
one
male
showed
a
slightly
increased
alkaline
phosphatase
level
at
30
days.
At
90
days,
one
male
showed
moderate
increases
in
glucose,
blood
urea
nitrogen
and
y­
glutamyl
transpeptidase
activity.
The
females
had
significant
increases
in
absolute
and
relative
liver
weights.
The
males
had
significant
decreases
in
absolute
and
relative
adrenal
weights,
absolute
thyroid/
parathyroid
weight
and
absolute
pituitary
weight.
The
biological
significance
of
the
changes
in
male
organ
weights
is
unclear
since
similar
changes
were
not
noted
in
higher
dose
groups.

At
necropsy,
treatment
related
gross
lesions
were
present
in
all
treated
groups
and
included
varying
degrees
of
discoloration
and/
or
enlargement
of
the
liver
and
discoloration
of
the
glandular
mucosa
of
the
stomach.
Histologic
examination
also
showed
lesions
in
all
treated
groups.
Centrilobular
to
midzonal
cytoplasmic
hypertrophy
of
hepatocytes
and
focal
necrosis
was
observed
in
the
liver;
the
incidence
and
relative
severity
were
greater
in
the
males.
In
addition,
especially
among
rats
in
the
300,
1000
and
3000
ppm
groups,
treatment
related
histologic
lesions
were
noted
in
the
primary
(
thymus,
bone
marrow)
and
secondary
(
spleen,
mesenteric
lymph
nodes)
lymphoid
organs,
stomach,
intestines,
muscle
and
skin.
In
the
thymus,
this
consisted
of
depletion
in
the
number
and
size
of
the
lymphoid
follicles
and
in
the
bone
marrow
hypocellularity
was
noted.
The
spleen
was
slightly
atrophied
with
a
corresponding
decrease
in
the
size
and
number
of
lymphoid
follicles
and
cells
and
a
similar
depletion
was
noted
in
the
mesenteric
lymph
nodes.
Mucosal
hyperkeratosis
and/
or
acanthosis
was
observed
in
the
forestomach
and
mucosal
hemorrhages
were
noted
in
the
glandular
portion
of
the
stomach.
Decrease
atrophy
in
the
height
and
thickness
of
the
villi
were
noted
in
the
small
intestine.
Atrophy
of
the
skeletal
muscle
was
noted,
as
well
as
epidermal
hyperkeratosis
and/
or
acanthosis
was
noted
in
the
skin.

CONCLUSIONS
Remarks:
Authors
conclusions
stated
above
in
results.
Reviewer
agrees.

REFERENCE
Goldenthal,
E.
I.,
D.
C.
Jessup,
R.
G.
Geil
and
J.
S.
Mehring.
1978.
Ninety­
day
subacute
rat
toxicity
study.
Study
No.
137­
085,
International
Research
and
Development
Corporation,
Mattawan,
MI.
ENV/
JM/
RD(
2002)
17/
FINAL
299
Title:
4­
Week
Capsule
Toxicity
Study
with
Perfluoroctane
Sulfonic
Acid
Potassium
Salt
(
PFOS;
T­
6295)
in
Cynomolgus
Monkeys.
Unaudited
Draft.
1998.

TEST
SUBSTANCE
Identity:
Potassium
Perfluorooctylsulfonate,
CAS
No.:
2795­
39­
3
Remarks:
T­
6295,
purity
not
specified.

METHOD
Method/
guideline
followed:
Range
finding
 
Unaudited
Draft
report,
sections
missing
Study
duration:
28
days
GLP
(
Y/
N):
Y,
report
contains
GLP
statement,
but
as
submitted,
unaudited
draft
report
with
sections
missing
Year
study
performed:
1998
Species/
Strain:
Monkey/
Cynomolgus
Sex:
both
Number
of
animals
per
dose:
2
per
sex,
0
mg/
kg/
day;
3
per
sex,
0.02
mg/
kg/
day;
1
per
sex,
2.0
mg/
kg/
day
Route
of
administration:
Capsule
in
stomach
Doses
tested
and
frequency:
0
mg/
kg/
day;
0.02
mg/
kg/
day;
2.0
mg/
kg/
day
Post­
observation
period:
none
Statistical
methods
used:
none
Remarks:
Monkeys
were
observed
at
least
daily
for
general
clinical
signs
and
body
weights
were
recorded
twice
weekly.
Hematological
and
clinical
chemistry
analyses
were
conducted
on
samples
collected
before
the
beginning
of
the
study
at
day
 
7
(
baseline
values)
and
day
29.
Additional
blood
samples
for
clinical
chemistry
were
collected
on
study
days
2,
7,
and
14.
Blood
samples
for
serum
PFOS
concentrations
were
taken
on
days
­
7,
2,
3,
7,
14,
and
29.
In
addition,
samples
from
day
­
7
and
day
29
were
analyzed
for
levels
of
estradiol,
estrone,
estriol,
thyroid
stimulating
hormone,
triiodothyronine,
and
thyroxin.
The
study
animals
were
terminated
as
scheduled
at
30
days.
At
necropsy
a
sample
of
liver
was
collected
from
each
animal
for
palmitoyl
CoA
oxidase
activity
analyses.
Samples
of
liver,
testes,
and
pancreas
were
collected
for
proliferation
cell
nuclear
antigen
evaluation.
A
sample
of
liver
was
also
collected
from
each
animal
for
PFOS
concentration
analysis.
The
following
organs
from
control
and
all
treated
groups
were
examined
microscopically:
adrenals,
eye,
kidney,
liver,
lung,
spleen,
pancreas,
femoral
bone
marrow,
testes,
and
thymus.
ENV/
JM/
RD(
2002)
17/
FINAL
300
RESULTS
NOAEL
(
dose
and
effect):
None
determined.

LOAEL
(
dose
and
effect):
None
determined
Toxic
response/
effects
by
dose
level:
None
determined
Statistical
results:
None,
high­
dose
too
few
animals
(
1
male,
1
female
Summary
Hormone
Analyses
Data
in
Males
on
Females
on
Days
 
7
and
29
Dose
Level
Sex
Day
Estradiol
pg/
mL
Estrone
pg/
mL
Estriol
pg/
mL
TSH
ØU/
mL
Triiodothyronine
ng/
dL
Thyroxin
Øg/
dL
0
Males
­
7
32.03
±
4.002
28.23
±
14.701
0.00
±
0.000
2.82
±
0.686
139.74
±
29.232
5.43
±
2.001
0
Males
29
32.09
±
0.205
21.93
±
4.30
0.00
±
0.000
2.15
±
2.008
186.35
±
6.935
4.32
±
1.407
0.02
Males
­
7
28.61
±
4.874
17.32
±
1.373
0.00
±
0.000
2.11
±
0.799
122.49
±
48.610
4.28
±
0.335
0.02
Males
29
34.25
±
7.998
14.72
±
4.767
0.00
±
0.000
3.10
±
0.584
188.34
±
26.229
4.19
±
0.612
2.0
Males
­
7
28.71
13.98
0.00
1.71
97.61
3.86
2.0
Males
29
18.16
20.12
0.00
0.95
90.73
3.18
0
Females
­
7
45.28
±
0.212
28.77
±
5.162
0.00
±
0.000
4.37
±
4.398
132.65
±
56.434
4.88
±
0.948
0
Females
29
53.47
±
20.082
25.97
±
2.128
0.00
±
0.000
2.95
±
0.530
162.70
±
29.366
4.26
±
0.438
0.02
Females
­
7
48.12
±
21.124
30.40
±
10.852
0.00
±
0.000
2.51
±
1.506
131.09
±
56.434
4.76
±
1.057
0.02
Females
29
58.72
±
27.628
31.61
±
0.921
0.00
±
0.000
2.76
±
1.764
180.58
±
15.203
4.05
±
1.108
2.0
Females
­
7
37.37
29.50
0.00
3.92
122.38
3.60
2.0
Females
29
19.82
25.85
0.00
90.17
3.38
CONCLUSIONS
Estradiol,
estrone,
thyroid
stimulating
hormone
(
TSH),
thyroxin,
and
triiodothyronine
levels
were
lower
in
the
high
dose
animals
at
the
end
of
the
study.
Since
the
numbers
of
tested
animals
are
small
(
one
male
and
one
female
in
the
high­
dose
group)
and
baseline
levels
are
variable,
it
is
not
clear
if
these
hormone
level
changes
are
treatment­
related.

Remarks:
None
REFERENCE
Thomford,
P.
J.
1998.
4­
Week
Capsule
Toxicity
Study
with
Perfluoroctane
Sulfonic
Acid
Potassium
Salt
(
PFOS;
T­
6295)
in
Cynomolgus
Monkeys.
Unaudited
Draft.
Study
No.
T­
6295.6,
for
3M,
St.
Paul,
MN,
by
Covance
Laboratories
Inc.,
Madison,
WI.
ENV/
JM/
RD(
2002)
17/
FINAL
301
Title:
26­
Week
Capsule
Toxicity
Study
with
Perfluorooctane
Sulfonic
Acid
Potassium
Salt
(
PFOS;
T6295)
in
Cynomolgus
Monkeys,
2002
TEST
SUBSTANCE
Identity:
Potassium
perfluorooctylsulfonate,
CAS
#
2795­
39­
3
Remarks:
FC­
95,
T­
6295,
purity
not
specified.

METHOD
Method/
guideline
followed:
No
guideline
followed.

Study
duration:
78
weeks,
26­
week
treatment
period,
followed
by
52
weeks
recovery.

GLP
(
Y/
N):
Y
Year
study
performed:
2000
Species/
strain:
Cynomolgus
monkeys
Sex:
Males
and
females
Number
of
animals
per
dose
group:
6
animals/
sex/
dose
group
for
groups
1,
3,
and
4;
4
animals/
sex
for
group
2.

Route
of
administration:
Oral
capsule
Doses
tested
and
frequency:
0
mg/
kg/
day,
0.03
mg/
kg/
day,
0.15
mg/
kg/
day,
or
0.75
mg/
kg/
day
Post­
observation
period:
52
weeks
Statistical
methods
used:
Levene's
test
for
variance
homogeneity:
ANOVA,
Dunnett's
t­
test
ANCOVA,
covariate­
adjusted
means,
5%
two­
tailed
probability
level.

Remarks:
Animals
were
observed
twice
daily
for
mortality
and
moribundity
and
were
examined
at
least
once
daily
for
abnormalities
and
signs
of
toxicity;
food
consumption
was
assessed
qualitatively.
Ophthalmic
examinations
were
done
before
initiation
of
treatment
and
during
weeks
26
and
52.
Body
weight
data
were
recorded
weekly
before
the
start
of
treatment,
on
Days
 
1
and
1
and
weekly
thereafter.
Blood
and
urine
samples
were
collected
for
clinical
hematology,
clinical
chemistry,
and
urinalysis
before
the
start
of
treatment
and
at
specified
intervals
during
treatment
and
recovery.
Blood
samples
were
also
taken
for
hormone
determinations.
Samples
of
serum
were
collected
at
various
time
points
during
the
study
and
sent
to
3M
for
analysis
of
PFOS
levels.
The
following
organs
were
weighed
at
scheduled
and
unscheduled
sacrifices;
paired
organs
were
weighed
separately:
adrenal
(
2),
brain,
epididymis
(
2),
kidney
(
2),
liver,
ovary
(
2),
pancreas,
testis
(
2),
and
thyroid
(
2)
with
parathyroid.
The
following
tissues
were
collected
for
histopathology:
adrenals
(
2),
aorta,
brain,
cecum,
cervix,
colon,
duodenum,
epididymis
(
2),
esophagus,
eyes
(
2),
femur
with
bone
marrow,
gallbladder,
heart,
ileum,
jejunum,
kidneys
(
2),
lesions,
liver,
lung,
mammary
gland,
mesenteric
lymph
node,
ovary
(
2),
pancreas,
pituitary,
prostate,
rectum,
salivary
gland
[
mandibular
(
2)],
sciatic
nerve,
seminal
vesicle
(
2),
skeletal
muscle
(
thigh),
skin,
spinal
cord
(
cervical,
thoracic,
and
lumbar),
spleen,
sternum
with
bone
marrow,
stomach,
testis
(
2),
thymus,
thyroid
(
2)
with
parathyroid,
trachea,
urinary
bladder,
uterus,
and
vagina.
Liver
specimens
from
the
0.15
and
0.75
ENV/
JM/
RD(
2002)
17/
FINAL
302
mg/
kg/
day
recovery
animals
were
collected
via
biopsy
and
analyzed
for
PFOS
levels.

Serum
and
liver
specimens
collected
from
test
animals
were
sent
to
the
3M
Laboratory
and
analyzed
for
the
presence
of
PFOS.
Serum
was
harvested
from
blood
that
was
centrifuged
within
one
hour
of
collection.
Liver
specimens
were
flash
frozen
in
liquid
nitrogen.
Both
liver
and
serum
samples
were
stored
in
a
freezer
set
to
maintain
specimens
at
­
60
to
 
80oC
until
shipped
to
the
3M
Lab.
Samples
were
shipped
frozen
and
on
dry
ice
from
Covance
Laboratories
to
3M
periodically
from
August
1998
through
March
2000
which
covered
the
in­
life
phase
of
the
study.
Once
received
at
3M
specimens
were
stored
in
freezer
at
either
 
55oC
±
10­
20oC
or
 
20oC
±
10oC.

During
the
first
26
weeks
of
the
study
a
total
of
550
serum
specimens
and
30
liver
specimens
were
collected.
Of
the
serum
specimens,
151
were
from
Group
1,
99
from
Group
2,
152
from
Group
3
and
148
from
Group
4.
Eight
liver
samples
were
collected
from
Group
1,
8
from
Group
2,
12
from
Group
3
and
14
from
Group
4.
In
the
recovery
Groups,
72
serum
and
4
liver
samples
were
collected
from
Group
1;
72
serum
and
4
liver
samples
from
Group
3
and
80
serum
and
4
liver
samples
from
Group
4.

Liver
and
serum
samples
were
extracted
using
an
ion­
pairing
reagent
and
methyl­
tert­
butyl
ether
(
MtBE).
Liver
samples
were
homogenized
prior
to
extraction.
Sample
extracts
were
analyzed
using
high­
pressure
liquid
chromatography­
electrospray/
tandem
mass
spectrometry
(
HPLC­
ES/
MS/
MS)
in
the
multiple
response
mode.
PFOS
levels
were
quantitated
by
external
standard
calibration.

Liver
samples
were
homogenized
in
water.
An
aliquot
of
each
liver
homogenate
and
all
serum
samples
were
spiked
with
THPFOS
and
extracted
using
an
ion­
pairing
extraction
procedure.
An
ion­
pairing
reagent
was
added
to
the
samples
and
the
ion
pairs
were
partitioned
into
MtBE.
The
extracts
were
evaporated
until
dry
on
a
nitrogen
evaporator
and
then
were
reconstituted
in
1.0
mL
of
methanol
and
passed
through
a
0.2
µ
m
nylon
filter.

The
analyses
were
performed
by
monitoring
one
or
more
product
ions
selected
from
a
single
primary
ion
characteristic
of
the
fluorochemical
of
interest
using
HPLC/
ES/
MS/
MS.
Molecular
ion
499,
the
primary
ion
for
PFOS
(
C8F17SO3
­)
analysis,
was
fragmented
to
produce
ion
99
(
FSO3
­).
Ion
99
was
monitored
for
quantitative
analysis.

RESULTS
NOAEL
(
dose
and
effect):
0.15
mg/
kg/
day.

LOAEL
(
dose
and
effect):
0.75
mg/
kg/
day.
Death,
liver
effects,
effect
on
cholesterol
Toxic
response/
effects
by
dose
level:
Death
at
0.75
mg/
kg/
day;
increased
absolute
liver
weight,
liver
to
body
weight
percentages,
liver
to
brain
weight
ratios
in
females
at
0.75
mg/
kg/
day;
absolute
and
relative
liver
weight;
Males
and
females
in
the
0.75
mg/
kg/
day
dose­
group
had
lower
total
cholesterol
and
lower
high
density
lipoprotein
cholesterol,
liver
organ
weights.

Statistical
results:
The
difference
in
weight
at
the
end
of
treatment
between
the
control
and
the
0.75
mg/
kg/
day
female
treatment
groups
was
statistically
significant;
the
effect
on
total
cholesterol
and
high
density
lipoprotein
cholesterol
in
the
0.75
mg/
kg/
dose
group
was
statistically
significant;
in
males
in
the
0.75
mg/
kg/
day
dose
group
the
liver
organ
weights
and
the
organ­
to
body
weight
percentages
were
statistically
significant
and
in
females
the
liver
weights,
the
organ­
to­
body
weight
percentage
and
the
organ­
brain
weight
ratio
were
all
significant.
ENV/
JM/
RD(
2002)
17/
FINAL
303
Remarks:

Males
weighed
3.3­
3.4
kg
and
females
weighed
2.8­
2.9
kg
at
the
beginning
of
the
study.
At
the
end
of
26
weeks
of
treatment,
males
weighed
3.7,
3.8,
3.5,
and
3.3
kg
for
the
0,
0.03,
0.15
and
0.75
mg/
kg/
day
treatment
groups
respectively.
Females
weighed
3.1,
3.1,
3.1
and
2.8
kg
for
the
0,
0.03,
0.15
and
0.75
mg/
kg/
day
treatment
groups
respectively.
The
difference
between
the
control
and
the
0.75
mg/
kg/
day
female
treatment
groups
was
statistically
significant.
At
the
end
of
the
recovery
period,
differences
in
weight
between
the
control
and
treated
animals
were
no
longer
obvious.

Two
males
from
the
0.75
mg/
kg/
day
group
did
not
survive
to
the
scheduled
sacrifice.
One
animal
died
after
dosing
on
Day
155
(
Week
23).
Clinical
signs
noted
in
this
animal
included:
constricted
pupils,
pale
gums,
few,
mucoid,
liquid
and
black­
colored
feces,
low
food
consumption,
hypoactivity,
labored
respiration,
dehydration,
and
recumbent
position.
In
addition,
the
animal
was
cold
to
the
touch.
An
enlarged
liver
was
detected
by
palpation.
Cause
of
death
was
determined
to
be
pulmonary
necrosis
with
severe
acute
inflammation.
On
day
179,
the
second
male
was
sacrificed
in
a
moribund
condition.
Clinical
signs
noted
included
low
food
consumption,
excessive
salivation,
labored
respiration,
hypoactivity
and
ataxia.
Cause
of
death
was
not
determined.

Males
and
females
in
the
0.75
mg/
kg/
day
dose­
group
had
lower
total
cholesterol
and
males
and
females
in
the
0.15
and
0.75
mg/
kg/
day
groups
had
lower
high
density
lipoprotein
cholesterol
during
treatment.
However,
only
the
effect
in
the
0.75
mg/
kg
group
was
statistically
significant.
The
effect
on
total
cholesterol
worsened
with
time.
By
day
182,
mean
total
cholesterol
for
males
and
females
in
the
high
dose
group
were
68%
and
49%
lower,
respectively,
that
than
levels
in
the
control
animals.
The
effect
on
high
density
lipoprotein
was
greater
than
that
seen
with
cholesterol.
On
day
182,
the
mean
high
density
lipoprotein
levels
were
79%
and
62%
lower
in
males
and
females,
respectively,
from
the
high
dose
group
than
they
were
in
male
and
female
control
animals.
Males
in
the
high
dose
group
also
had
lower
total
bilirubin
concentrations
and
higher
serum
bile
acid
concentrations
than
males
in
either
the
control
or
other
treatment
groups.
The
effect
on
total
cholesterol
was
reversed
within
5
weeks
of
recovery
and
the
effect
on
high
density
lipoprotein
cholesterol
was
reversed
within
9
weeks
of
recovery.

Estradiol
values
were
lower
in
males
given
0.75
mg/
kg/
day
on
days
62,
91,
and
182
by
because
of
variation
only
the
day
182
value
was
significant.
Estrone
values
were
generally
higher
in
the
treated
females
on
days
37
62
and
91
by
again
because
of
variation
in
the
data
none
of
these
values
were
significantly
different.
Triiodothyronine
values
were
notably
lower
on
days
91
and
182
in
males
and
females
given
0.15
and
0.75
mg/
kg/
day.
There
were
other
instances
in
which
hormone
values
in
treated
groups
were
different
from
those
of
controls
but
these
differences
were
not
consistent
over
time
or
between
sexes,
were
not
clearly
dose­
related
and
did
not
appear
to
be
related
to
the
administration
of
the
test
material.
Apparent
differences
in
the
sexual
maturity
of
both
males
and
females
used
in
the
study
complicates
the
interpretation
of
the
hormone
data.

At
terminal
sacrifice,
females
in
the
0.75
mg/
kg/
day
dose­
group
had
increased
absolute
liver
weight,
liverto
body
weight
percentages,
and
liver­
to­
brain
weight
ratios.
In
males,
liver­
to
body
weight
percentages
were
increased
in
the
high­
dose
group
compared
to
the
controls.
"
Mottled"
livers
were
observed
in
two
high­
dose
males
and
in
one
high­
dose
female.
Of
the
two
males
not
surviving
until
the
scheduled
terminal
sacrifice,
one
had
a
"
mottled"
and
large
liver.
Three
of
4
high­
dose
males
(
including
those
that
did
not
survive
to
scheduled
sacrifice)
had
centrilobular
or
diffuse
hepatocellular
hypertrophy
that
was
also
observed
in
all
high­
dose
females.
Centrilobular
or
diffuse
hepatocellular
vacuolation
occurred
in
2
of
4
females
and
2
of
4
males
in
the
high­
dose
group.

No
PFOS
related
lesions
were
observed
at
recovery
sacrifice
indicating
that
the
effects
seen
at
terminal
sacrifice
may
be
reversible.
ENV/
JM/
RD(
2002)
17/
FINAL
304
Although
low
levels
of
PFOS
were
often
detected
in
the
sera
and
liver
of
the
control
animals,
these
levels
were
significantly
lower
than
those
found
in
the
low
dose
test
animals.
PFOS
levels
in
the
sera
of
test
animals
increased
with
dose
during
treatment
from
21.0
±
1.57
and
20.4
±
2.71
µ
g/
ml
in
the
Group
4
males
and
females
respectively
at
the
end
of
Week
1
to
194
±
8.94
and
160
±
23.1
µ
g/
ml
in
males
and
females
respectively
in
Group
4
at
the
end
of
Week
27.
During
recovery,
PFOS
levels
in
serum
samples
decreased
over
time
until
they
reached
41.1
±
25.9
µ
g/
ml
in
males
and
41.4
±
1.15
µ
g/
ml
in
females
from
Group
4
at
79
weeks
post­
treatment.
Control
values
were
<
LOQ
(
the
limit
of
quantitation)
at
Week
4
in
both
males
and
females
and
0.0215
±
0.00296
and
0.0243
±
0.00355
µ
g/
ml
in
males
and
females
respectively
at
the
end
of
Week
79.
The
serum
values
for
selected
weeks
of
treatment
and
recovery
are
shown
in
the
table
below.
There
were
no
significant
differences
between
PFOS
levels
in
the
sera
of
treated
males
and
females.

Average
PFOS
Concentrations
(
µ
g/
ml)
in
Serum
of
Monkeys
for
Selected
Weeks
During
Treatment
and
Recovery
Group
1
0.0
mg/
kg/
day
Group
2
0.03
mg/
kg/
day
Group
3
0.15
mg/
kg/
day
Group
4
0.75
mg/
kg/
day
Males
Females
Males
Females
Males
Females
Males
Females
Week
1
<
LOQ
<
LOQ
0.869
±
0.147
0.947
±
0.110
4.60
±
0.782
3.71
±
0.455
21.0
±
1.57
20.4
±
2.71
Week
4
<
LOQ
<
LOQ
3.20
±
0.577
3.40
±
0.291
17.8
±
1.68
16.5
±
1.87
95.3
±
70.4
92.7
±
39.6
Week
16
0.0407
±
0.0110
0.0432
±
0.0081
11.2
±
2.44
10.5
±
1.90
56.2
±
5.84
42.1
±
4.04
189
±
15.9
162
±
19.3
Week
27
0.0529
±
0.0145
0.0416
±
0.0148
15.9
±
5.54
11.1
±
1.52
68.1
±
5.75
58.5
±
4.67
194
±
8.93
160
±
23.9
Week
35
0.0459
±
0.00303
0.0723
±
0.00352
Not
Determined
Not
Determined
84.5
±
12.0
74.7
±
9.53
181
±
19.5
171
±
10.1
Week
47
0.0355
±
0.00221
0.0459
±
0.00323
Not
Determined
Not
Determined
48.3
±
3.69
42.6
±
6.70
124
±
25.9
98.3
±
8.32
Week
57
0.0327
±
000526
0.0445
±
0.00385
Not
Determined
Not
Determined
30.2
±
2.36
32.3
±
1.34
78.0
±
16.3
106
±
3.84
Week
69
0.0406
±
0.00313
0.0400
±
0.00301
Not
Determined
Not
Determined
26.4
±
2.59
34.5
±
3.46
84.0
±
52.4
75.0
±
5.25
Week
79
0.0215
±
0.00296
0.0243
±
0.00355
Not
Determined
Not
Determined
19.1
±
0.805
21.4
±
2.01
41.1
±
25.9
41.4
±
1.15
LOQ
=
Lowest
Observable
Concentration
Liver
values
behaved
in
a
manner
similar
to
serum
values
and
increased
over
time.
At
Week
27
mean
ENV/
JM/
RD(
2002)
17/
FINAL
305
PFOS
values
on
an
RSD
basis
were
22.2
±
0.0269
in
Group
1
males
and
16.8
±
0.0178
in
females
in
Group
1;
27.0
±
4.66
and
9.73
±
2.15
in
males
and
females
in
Group
2;
33.1
±
19.5
in
males
and
21.4
±
14.9
in
females
in
Group
3,
and
6.03
±
23.9
in
males
and
5.00
±
13.6
in
females
in
Group
4.
At
Week
79
values
in
the
liver
were
71.0
±
33.4
in
males
and
21.4
±
10.8
in
females
in
Group
4.
At
Week
80,
values
were
14.9
±
1.38
in
Group
3
males
and
23.5
±
4.98
in
Group
3
females.

CONCLUSIONS
PFOS
is
toxic
to
cynamolgous
monkeys
at
0.75
mg/
kg/
day
causing
death,
alterations
in
total
cholesterol,
and
effecting
liver
weight
and
causing
hepatocellular
hypertrophy
and
vacuolation
in
both
treated
males
and
females.
However,
the
effects
on
cholesterol
and
the
liver
appear
to
be
reversible
after
a
52
week
recovery
period.

REFERENCE
Thomford,
PJ.
(
2000).
26­
Week
Capsule
Toxicity
Study
with
Perfluorooctane
Sulfonic
Acid
Potassium
Salt
(
PFOS;
T6295)
in
Cynomolgus
Monkeys.
Unaudited
Draft
Final
Report
Prepared
for
3M,
St
Paul,
Minnesota
by
Covance
Laboratories,
Inc.,
Madison
Wisconsin
53704­
2595.
April
12,
2000.
502
pp.

Seacat,
AM.
Analytical
Laboratory
Report
from
the
26­
Week
Capsule
Toxicity
Study
with
Perfluoroctanesulfonic
Acid
Potassium
Salt
(
T­
6295)
in
Cynomolgus
Monkeys
on
the
Determination
of
the
Presence
and
Concentration
of
Perfluorooctanesulfonate
(
PFOS)
in
Liver
and
Serum
Samples.
3M
Medical
Department
Study:
T­
6295.7;
Covance
In­
Life
Study:#
6329­
223.
Analytical
Study;
FACT
TOX­
030;
3M
Laboratory
Request
No.
U2279.
ENV/
JM/
RD(
2002)
17/
FINAL
306
DEVELOPMENTAL
TOXICITY
STUDIES
Title:
Oral
Teratology
Study
of
FC­
95
in
Rats
­
Experiment
No.
0680TR0008
TEST
SUBSTANCE
Identity:
Potassium
Perfluorooctylsulfonate,
CAS
No.
2795­
39­
3
Remarks:
FC­
95,
Lot
640.

METHOD
Method/
Guideline
followed
(
i.
e.,
OECD
414,
etc.):
Actual
guideline
followed
was
not
specified
but
appears
to
be
similar
in
design
to
OECD
414.

GLP
(
Y/
N):
The
procedure
complies
with
the
general
recommendations
of
the
FDA
issued
in
January,
1966
("
Guidelines
for
Reproduction
Studies
for
Safety
Evaluation
of
Drugs
for
Human
Use").
The
study
was
conducted
according
to
the
1978
Good
Laboratory
Practice
regulations
and
Safety
Evaluation
Laboratory's
Standard
Operating
Procedures.

Year
study
performed:
1980
Species/
Strain:
Sprague­
Dawley
rats
Number
of
animals
per
dose:
22
Route
of
administration:
Gavage
Dosing
regimen
(
list
all
with
units):
Four
groups
of
22
time­
mated
Sprague­
Dawley
rats
were
administered
Potassium
Perfluorooctylsulfonate
in
corn
oil
by
gavage
on
gestation
days
6­
15.
Doses
were
adjusted
according
to
the
most
recent
recorded
body
weight.

Doses:
0,
1,
5,
and
10
mg/
kg/
day
Statistical
methods
used:
The
animals
will
be
assigned
cages
according
to
a
computer­
generated
random
numbers
table.
The
statistical
methods
to
be
used
for
analysis
of
the
data
are:
Dunnett's
t
test
for
dam
and
pup
weights,
number
of
fetuses,
number
of
resorption
sites,
number
of
implantation
sites
and
number
of
corpora
lutea;
Chi
square
for
percent
abnormalities.

Remarks
 
Detail
and
discuss
any
significant
protocol
parameters
and
deviations:
Potassium
Perfluorooctylsulfonate
was
administered
in
corn
oil
by
gavage
to
four
groups
of
22
time­
mated
Sprague­
Dawley
rats
weighing
175­
261g,
at
doses
of
0,
1,
5,
and
10
mg/
kg/
day
PFOS
on
days
6­
15
of
gestation
(
Gortner,
1980).
Purina
Laboratory
Chow
and
water
were
available
ad
libitum.
The
animals
were
dosed
according
to
a
constant
dose
volume
of
5
ml/
kg
of
body
weight
and
observed
daily
from
day
3
through
day
20
of
gestation
for
abnormal
clinical
signs.
Body
weights
were
recorded
on
days
3,
6,
9,
12,
15,
and
20
of
gestation
and
the
rats.
All
animals
were
sacrificed
on
day
20
by
cervical
dislocation
and
the
ovaries,
uteri
and
contents
were
examined
for
the
number
of
corpora
lutea,
number
of
viable
and
non­
viable
fetuses,
number
of
resorption
sites,
and
number
of
implantation
sites.
Fetuses
were
weighed
and
sexed
and
subjected
to
external
gross
necropsy.
Approximately
one­
third
of
the
fetuses
were
fixed
in
Bouin's
solution
and
examined
for
visceral
abnormalities
by
free­
hand
sectioning
by
the
Wilson
technique.
The
remaining
fetuses
were
subjected
to
a
ENV/
JM/
RD(
2002)
17/
FINAL
307
skeletal
examination
using
alizarin
red.

RESULTS
NOAEL
(
dose
and
effect)
 
maternal
and
developmental:
A
NOAEL
of
5
mg/
kg/
day
for
maternal
toxicity
was
indicated.
No
signs
of
maternal
toxicity
were
reported
at
doses
of
5
mg/
kg/
day
and
below.
A
NOAEL
for
developmental
toxicity
could
not
be
established;
signs
of
developmental
toxicity
were
evident
at
all
doses.

LOAEL
(
dose
and
effect)
 
maternal
and
developmental:
A
LOAEL
of
10
mg/
kg/
day
for
maternal
toxicity
was
indicated
based
on
significant
reductions
in
mean
body
weights
during
gestation
day
12­
20
at
the
high­
dose
group
of
10
mg/
kg/
day.
A
LOAEL
of
1
mg/
kg/
day
for
developmental
toxicity
was
indicated
based
on
abnormalities
of
the
lens
of
the
eye.

Toxic
response/
effects
by
dose
level
­
maternal:
Significant
reductions
in
mean
body
weights
during
GD
12­
20
at
the
high­
dose
group
of
10
mg/
kg/
day.

Toxic
response/
effects
by
dose
level
­
developmental:
Unusually
high
incidences
of
developmental
variations
and
abnormalities
of
the
lens
of
the
eye
were
observed
in
all
dose
groups.

Statistical
results:
At
10
mg/
kg/
day,
mean
maternal
body
weights
were
statistically
significantly
lower
than
controls
(
Dunnett's
test
p
<
0.05).
Mean
litter
data
and
pup
weights
were
not
significantly
different
from
controls
(
Dunnett's
t
test
p<
0.05).
Number
of
fetuses
with
gross
findings
were
not
significantly
different
from
controls
(
Chi­
square
p<
0.05).
Number
and
percent
of
fetuses
with
skeleton
findings
were
not
significantly
different
from
controls
(
Chisquare
p<
0.05).
Number
and
percent
of
fetuses
with
internal
findings
­
­
developmental
lens
abnormalities
with
secondary
lens
aberrations
were
significantly
higher
than
controls
(
Chi­
square
p<
0.05).

Remarks
 
Additional
information
to
adequately
assess
the
data:
Signs
of
maternal
toxicity
consisted
of
significant
reductions
in
mean
body
weights
during
GD
12­
20
at
the
high­
dose
group
of
10
mg/
kg/
day.
No
other
signs
of
maternal
toxicity
were
reported.
Developmental
toxicity
evident
at
doses
of
10
mg/
kg/
day
consisted
of
reductions
in
the
mean
number
of
implantation
sites,
corpora
lutea,
resorption
sites
and
the
mean
numbers
of
viable
male,
female,
and
total
fetuses,
but
the
differences
were
not
statistically
significant.
In
addition,
unusually
high
incidences
of
unossified,
assymetrical,
bipartite,
and
missing
sternebrae
were
observed
in
all
dose
groups;
however,
these
skeletal
variations
were
also
observed
in
control
fetuses
at
the
same
rate
and
therefore
were
not
considered
to
be
treatment­
related.
The
most
notable
sign
of
developmental
toxicity
observed
in
all
dose
groups
consisted
of
abnormalities
of
the
lens
of
the
eye,
which
was
not
seen
in
controls.
The
proportion
of
fetuses
with
the
lens
abnormality
in
one
or
both
lenses
was
significantly
higher
in
the
high
dose
group.
All
eye
abnormalities
appeared
to
be
localized
to
the
area
of
the
embryonal
lens
nucleus,
although
a
variety
of
morphological
appearances
were
present
within
that
location.
According
to
the
authors,
this
abnormality
appeared
to
be
an
arrest
in
development
of
the
primary
lens
fibers
forming
the
embryonal
lens
nucleus.
Secondary
lens
fiber
development
progressed
normally
except
immediately
surrounding
the
abnormal
embryonal
nucleus.
An
amendment
to
the
results
and
discussion
section
concludes
that
the
gross
finding
of
a
lens
cleft
was
an
artifact
created
by
freehand
sectioning
and
the
range
of
gross
lens
observations
and
the
differences
among
the
dose
group
incidences
were
due
to
the
maner
and
frequency
in
which
the
lens
cleft
artifact
was
created
by
free­
hand
sectioning
and
the
limitations
inherent
in
visualizing
the
embryonal
nucleus.
Additionally,
a
subsequent
study
(
Wetzel,
1893)
of
similar
design
was
not
able
to
repeat
this
finding.
ENV/
JM/
RD(
2002)
17/
FINAL
308
CONCLUSIONS
Given
the
explanation
that
the
eye
abnormalities
reported
at
doses
as
low
as
1
mg/
kg/
day
in
this
study
may
have
been
due
to
an
artifact
of
sectioning
and
that
these
findings
could
not
be
repeated
in
a
second
study
of
similar
design,
it
is
entirely
plausible
that
the
lens
defect
observed
in
this
study
is
not
treatment­
related.

REFERENCE
Gortner,
E.
G.
1980.
Safety
Evaluation
Laboratory
and
Riker
Laboratories,
Inc.
Experiment
Number:
0680TR0008,
December,
1980.
"
Oral
Teratology
Study
of
FC­
95
in
Rats".
ENV/
JM/
RD(
2002)
17/
FINAL
309
Title:
Rat
Teratology
Study
T­
3351
Final
Report
 
Project
No.
154­
160
TEST
SUBSTANCE
Identity:
Potassium
Perfluorooctylsulfonate,
CAS
No.
2795­
39­
3
Remarks:
T­
3351,
Lot
No.
80275
METHOD
Method/
Guideline
followed
(
i.
e.,
OECD
414,
etc.):
Actual
guideline
followed
was
not
specified
but
appears
to
be
similar
in
design
to
OECD
414.

GLP
(
Y/
N):
Quality
Assurance
inspections
of
the
study
and
review
of
the
final
report
were
conducted
according
to
the
standard
operating
procedures
of
the
Office
of
Quality
Assurance
and
according
to
the
general
requirements
of
the
Good
Laboratory
Practice
regulations
that
were
issued
on
December
22,
1978,
by
the
Food
and
Drug
Administration
for
compliance
on
and
after
June
20,
1979.

Year
study
performed:
1983
Species/
Strain:
Sprague­
Dawley
rats
Number
of
animals
per
dose:
25
Route
of
administration:
Gavage
Dosing
regimen
(
list
all
with
units):
Four
groups
of
25
pregnant
Sprague­
Dawley
rats
were
administered
Potassium
Perfluorooctylsulfonate
in
corn
oil
by
gavage
on
gestation
days
6­
15.
Doses
were
adjusted
according
to
the
most
recent
recorded
body
weight.

Doses:
0,
1,
5,
and
10
mg/
kg/
day
Statistical
methods
used:
Statistical
methods
used
for
analysis
of
the
data
:
Dunnett's
t
test
for
control
vs.
compound­
treated
group
mean
comparisons.
If
the
variances
were
proved
to
be
homogeneous,
the
data
were
analysed
by
one­
way
classification
analysis
of
variance
(
ANOVA).
Mean
fetal
body
weights
per
litter
were
statistically
analysed
as
follows:
Bartlett's
test
for
homogeneity
of
variances
was
performed
by
oneway
classification
of
covariance
(
ANCOVA).

If
ANCOVA
was
significant,
control
vs.
treatment
group
comparisons
were
analysed
using
the
Games
and
Howell
modification
of
the
Tukey­
Kramer
honestly
significant
difference
test.
Tests
for
homogeneity
of
variances,
ANOVA,
and
ANCOVA
were
evaluated
at
the
5%
one­
tailed
probability
level.
Control
vs.
compound­
treated
group
mean
comparisons
of
the
above
data
were
evaluated
at
the
5%
two­
tailed
probability
level.
Percent
fetal
viability,
percent
fetal
loss
(
dead
and
resorbing
fetuses),
percent
early,
late,
and
total
resorptions,
and
the
number
of
dead
fetuses
were
analysed
by
nonparametric
one­
way
ANOVA
and
the
Terpstra­
Jonckheere
test
for
trend.
The
litter
was
used
as
the
experimental
unit.
Teratology
data
were
analysed
using
the
Cochran­
Armitage
test
for
linear
trend
in
proportions.
If
a
significant
trend
was
noted,
the
results
of
Fisher's
"
exact"
test
were
evaluated
at
the
one­
tailed,
5%
level.
If
a
significant
trend
was
not
observed,
or
if
there
was
a
significant
trend
with
severe
departure
from
it,
the
results
of
Fisher's
"
exact"
test
were
evaluated
at
the
two­
tailed,
5%
level.
ENV/
JM/
RD(
2002)
17/
FINAL
310
Remarks
 
Detail
and
discuss
any
significant
protocol
parameters
and
deviations:
Potassium
perfluorooctylsulfonate
was
administered
in
corn
oil
by
gavage
to
four
groups
of
25
pregnant
Sprague­
Dawley
rats
at
doses
of
0,
1,
5,
and
10
mg/
kg/
day
PFOS
on
gestation
days
(
GD)
6­
15
(
Wetzel,
1983).
Sexually
mature
Sprague­
Dawley
rats,
one
per
sex
per
cage,
were
paired
until
confirmation
of
mating
or
until
two
weeks
had
elapsed.
Mating
was
confirmed
by
daily
vaginal
examinations
for
the
presence
and
viability
of
sperm
or
the
presence
of
a
copulatory
plug.
The
day
of
confirmation
of
mating
was
designated
as
day
0
of
gestation.
Purina
Rodent
Laboratory
Chow
5001
and
tap
water
were
available
ad
libitum.
A
dose
volume
of
3
ml/
kg
of
body
weight
was
administered
and
doses
were
adjusted
according
to
the
most
recently
recorded
body
weight
measurements.
Dams
were
observed
twice
daily
for
signs
of
mortality
and
moribundity
and
once
daily
for
clinical
signs
of
toxicity.
Individual
body
weights
and
food
consumption
were
recorded
on
GD
6,
8,
12,
16,
and
20.
Animals
were
sacrificed
on
GD
20
by
CO2
asphyxiation
and
the
fetuses
were
delivered
by
cesarean
section
on
GD
20.
A
gross
necropsy
was
performed
on
all
dams.
The
uterus
from
each
female
was
excised,
weighed
and
examined
for
the
number
and
placement
of
implantation
sites,
number
and
of
live
and
dead
fetuses,
number
of
early
and
late
resorptions,
and
any
abnormalities
and
then
weighed
again
after
the
contents
were
removed.
The
ovaries
were
examined
for
the
number
of
corpora
lutea.
Each
female
was
examined
by
gross
necropsy.
Each
fetus
was
sexed,
weighed,
and
examined
externally.
Approximately
one­
third
of
the
fetuses
were
fixed
in
Bouin's
solution
and
examined
for
visceral
abnormalities
by
the
Wilson
technique,
with
particular
attention
to
the
eyes,
palate,
and
brain.
The
remaining
fetuses
were
subjected
to
a
skeletal
examination
that
included
evaluation
of
the
skull,
long
bones,
vertebral
column,
rib
cage,
extremities,
and
pectoral
and
pelvic
girdles
using
alizarin
red;
bone
alignment
and
degree
of
ossification
were
also
evaluated.

RESULTS
NOAEL
(
dose
and
effect)
 
maternal
and
developmental:
The
NOAEL
for
maternal
toxicity
is
1
mg/
kg/
day.
The
NOAEL
for
developmental
toxicity
is
1
mg/
kg/
day.

LOAEL
(
dose
and
effect)
 
maternal
and
developmental:
The
LOAEL
for
maternal
toxicity
is
5
mg/
kg/
day,
based
on
clinical
signs
of
toxicity,
decreases
in
body
weight
and
food
consumption,
decreases
in
uterine
weights,
and
an
increased
incidence
in
gastrointestinal
lesions.

The
LOAEL
for
developmental
toxicity
is
5
mg/
kg/
day,
based
on
decreased
fetal
body
weight
and
increases
in
external
and
visceral
anomalies
and
variations.

Toxic
response/
effects
by
dose
level
­
maternal:
Clinical
signs
of
toxicity,
decreases
in
body
weights
and
food
consumption
at
5
and
10
mg/
kg/
day;
decreases
in
uterine
weights,
increased
incidence
in
gastrointestinal
lesions,
and
two
deaths
at
10
mg/
kg/
day.

Toxic
response/
effects
by
dose
level
­
developmental:
Decreased
fetal
weight
at
5
and
10
mg/
kg/
day;
external
and
visceral
anomalies
and
skeletal
variations
at
10
mg/
kg/
day.

Statistical
results:
Statistically
significant
differences
between
controls
and
treated
were
noted
for
the
following
maternal
endpoints:
mean
body
weight
gain,
mean
total
food
consumption,
and
mean
gravid
uterine
weight.
Nonparametric
analysis
of
the
mean
incidence
of
late
resorptions,
total
resorptions,
number
of
dead
fetuses,
and
fetal
loss
did
not
indicate
statistical
significance;
however,
there
was
a
significant
linear
trend
towards
an
increased
incidence
in
these
data
with
respect
to
control.
The
primary
trend
component
was
contributed
by
the
high­
dose
group.
Statistically
significant
treatment­
related
increases
in
the
incidences
of
visceral
anomalies
and
skeletal
variants
were
also
observed.
ENV/
JM/
RD(
2002)
17/
FINAL
311
Remarks
 
Additional
information
to
adequately
assess
the
data:
Evidence
of
maternal
toxicity,
that
was
observed
at
the
5
and
10
mg/
kg/
day
dose
groups
both
during
and
following
treatment
and
considered
to
be
treatment­
related,
consisted
of
hunched
posture,
anorexia,
bloody
vaginal
discharge,
uterine
stains,
alopecia,
rough
haircoat,
and
bloody
crust.
Significant
decreases
in
mean
body
weight
gains
during
GD
6­
8,
6­
16,
and
0­
20
were
also
observed
at
the
5
and
10
mg/
kg/
day
dose
groups.
These
reductions
were
considered
to
be
treatment­
related
since
mean
body
weight
gains
were
greater
than
controls
during
the
post­
exposure
period
(
GD
16­
20).
Significant
decreases
in
mean
total
food
consumption
were
observed
on
GD
17­
20
in
the10
mg/
kg/
day
dose
group,
and
on
GD
7­
16
and
0­
20
in
both
the
5
and
10
mg/
kg/
day
dose
groups.
The
mean
gravid
uterine
weight
in
the
10
mg/
kg/
day
dose
group
was
significantly
lower
when
compared
with
controls.
The
mean
terminal
body
weights
minus
the
gravid
uterine
weights
were
lower
in
all
treated
groups,
with
significant
decreases
at
5
and
10
mg/
kg/
day.
High­
dose
animals
also
exhibited
an
increased
incidence
in
gastrointestinal
lesions.
No
significant
differences
were
observed
in
pregnancy
rates,
number
of
corpora
lutea,
and
number
and
placement
of
implantation
sites
among
treated
and
control
groups.
Two
dams
in
the
10
mg/
kg/
day
dose
group
were
found
dead
on
GD
17.

Signs
of
developmental
toxicity
included
a
dose­
related
trend
toward
an
increased
incidence
of
late
resorptions,
total
resorptions,
number
of
dead
fetuses,
and
fetal
loss,
although,
none
of
these
effects
were
statistically
significantly
different
from
controls.
Significant
decreases
in
mean
fetal
weights
for
both
males
and
females
were
observed
in
the
5
and
10
mg/
kg/
day
dose
groups.
The
percent
of
male
fetuses
was
52%,
54%,
and
60%
for
1,
5,
and
10
mg/
kg/
day,
respectively,
compared
to
44%
in
controls.
Statistically
significant
increases
in
incomplete
closure
of
the
skull
were
observed
in
the
low­
and
high­
dose
groups
but
not
in
the
mid­
dose
group.
Statistically
significant
increases
in
the
incidences
in
the
number
of
litters
containing
fetuses
with
visceral
anomalies,
delayed
ossification,
and
skeletal
variations
were
observed
in
the
high
dose
group
of
10
mg/
kg/
day.
These
included
external
and
visceral
anomalies
of
the
cleft
palate,
subcutaneous
edema,
and
cryptorchsm
as
well
as
delays
in
skeletal
ossification
of
the
skull,
pectoral
girdle,
rib
cage,
vertebral
column,
pelvic
girdle
and
limbs.
Skeletal
variations
in
the
ribs
and
sternebrae
were
also
observed
at
this
dose
level.

CONCLUSIONS
The
developmental
eye
abnormalities
that
were
seen
in
Gortner
(
1980)
were
not
observed
in
the
present
developmental
toxicity
study
even
though
the
study
design
and
doses
were
the
same.
Findings
of
abnormalities
in
eye
development
were
initially
thought
to
be
treatment­
related
but
then
later
suggested
as
being
artifacts
of
sectioning.

REFERENCE
Wetzel,
L.
T.
1983.
Hazelton
Laboratories
America,
Inc.
Project
Number:
154­
160,
December
19,
1983.
"
Rat
Teratology
Study,
T­
3351,
Final
Report".
ENV/
JM/
RD(
2002)
17/
FINAL
312
Title:
ORAL
(
STOMACH
TUBE)
DEVELOPMENTAL
TOXICITY
STUDY
OF
PFOS
IN
RABBITS
 
3M
T­
6295.10,
ARGUS
RESEARCH
LABORATORIES
STUDY
NUMBER:
6295.10,
1999.

TEST
SUBSTANCE
Identity:
Potassium
Perfluorooctylsulfonate
(
PFOS),
CAS
No.
2795­
39­
3
Remarks:
PFOS
 
Lot
217,
98.4%
pure
(
SMD
Analytical
Request
53030)
Analytical
Documentation
filed
along
with
final
report.
Note:
Same
lot
as
used
in
two­
year
rat
PFOS
carcinogenicity
study
(
T­
6295,
Covance
6329­
183).

METHOD
Method/
Guideline
followed
(
i.
e.,
OECD
414,
etc.):
The
requirements
of
the
International
Conference
on
Harmonization
(
ICH)
Harmonized
Tripartite
Guideline
on
Detection
of
Toxicity
to
Reproduction
for
Medicinal
products,
stages
C
and
D
of
the
reproductive
process
in
a
non­
rodent
species
were
used
as
the
basis
for
study
design
(
U.
S.
Food
and
Drug
Administration,
1994.
Federal
Register,
September
22,
1194,
Vol.
59,
No.
183).

GLP
(
Y/
N):
The
study
was
conducted
in
compliance
with
the
Good
Laboratory
Practice
(
GLP)
regulations
of
the
U.
S.
Food
and
Drug
Administration
(
FDA),
the
Japanese
Ministry
of
Health
and
Welfare
(
MHW)
and
the
European
Economic
Community
(
EEC).
There
were
no
significant
deviations
from
the
GLP
regulations
that
affected
the
quality
or
integrity
of
the
study.

Year
study
performed:
1999
Species/
Strain:
New
Zealand
White
rabbits
Number
of
animals
per
dose:
22
(
additional
total
of
19
rabbits,
3­
5
per
dose,
were
included
in
a
satellite
study
to
determine
the
concentrations
of
PFOS
in
maternal
liver
and
serum.)

Route
of
administration:
Gavage
Dosing
regimen
(
list
all
with
units):
Four
groups
of
22
pregnant
New
Zealand
White
rabbits
were
administered
Potassium
Perfluorooctylsulfonate
(
PFOS)
in
0.5%
Tween­
80
by
gavage
on
gestation
days
7­
20.
A
dose
volume
of
5
ml/
kg
was
administered,
adjusted
daily
on
the
basis
of
individual
body
weights.

Doses:
0,
0.1,
1.0,
2.5,
and
3.75
mg/
kg/
day
(
In
the
satellite
study
the
number
of
does
was
3,
5,
3,
3
and
5
in
the
control,
0.1,
1.0,
2.5
and
3.75
mg/
kg/
day
groups,
respectively).

Statistical
methods
used:
The
animals
will
be
assigned
to
individual
housing
on
the
basis
of
computer­
generated
random
units.
The
litter
was
the
unit
of
measurement.
Clinical
observation
and
other
proportion
data
were
analysed
using
the
Variance
Test
for
Homogeneity
of
the
Binomial
Distribution.
Continuous
data
(
e.
g.,
maternal
body
weights,
body
weight
changes,
feed
consumption
values
and
litter
averages
for
percent
male
fetuses,
percent
resorbed
conceptuses,
fetal
body
weights,
fetal
anomaly
data
and
fetal
ossification
site
data)
were
analyzed
using
Bartlett's
Test
of
Homogeneity
of
Variances
and
the
Analysis
of
Variance.
If
the
Analysis
of
Variance
was
significant,
Dunnett's
Test
was
used
to
identify
the
statistical
significance
of
the
individual
groups.
If
the
Analysis
of
Variance
was
not
appropriate,
theKruskal­
Wallis
Test
was
used.
In
cases,
in
which
Kruskal­
Wallis
Test
was
statistically
significant
(
p<
0.05),
Dunn's
Method
of
Multiple
Comparisons
was
used
to
identify
the
statistical
significance
of
the
individual
groups.
Count
data
obtained
ENV/
JM/
RD(
2002)
17/
FINAL
313
at
Caesarean­
sectioning
were
evaluated
using
the
procedures
described
for
the
Kruskal­
Wallis
Test.

Remarks
 
Detail
and
discuss
any
significant
protocol
parameters
and
deviations:
Timed­
pregnant
New
Zealand
White
rabbits
(
obtained
from
Covance
Research
Products,
Inc.),
22
per
group,
were
given
doses
of
0,
0.1,
1.0,
2.5
or
3.75
mg/
kg/
day
PFOS
in
0.5%
Tween­
80
by
gavage
on
gestation
days
7­
20.
A
dose
volume
of
5
mL/
kg
was
administered,
adjusted
daily
on
the
basis
of
individual
body
weights.
The
does
were
observed
twice
daily
for
viability,
and
clinical
observations
were
recorded
1
hour
prior
to
and
after
dosing
during
the
treatment
period
and
once
daily
during
the
post­
treatment
period
(
i.
e.
gestation
days
20­
29).
Maternal
body
weights
were
recorded
on
gestation
days
0
and
6­
29;
food
consumption
was
recorded
daily
throughout
the
study.
On
gestation
day
29,
the
does
were
euthanized;
a
gross
necropsy
of
the
thoracic,
abdominal
and
pelvic
viscera
was
conducted
and
the
number
of
corpora
lutea
in
each
ovary
was
recorded.
The
uteri
were
examined
for
number
and
distribution
of
implantations,
live
and
dead
fetuses,
and
early
and
late
esorptions.
The
fetuses
were
weighed,
sexed
and
examined
for
external
abnormalities.
All
fetuses
were
examined
for
visceral
and
skeletal
abnormalities
and
the
brain
of
one­
half
of
the
fetuses
were
free­
hand
cross­
sectioned
and
examined
in
situ.
In
the
satellite
study,
the
does
were
euthanized
on
gestation
day
21,
blood
samples
were
collected,
and
the
liver
was
weighed
and
sectioned.
The
fetuses
were
removed
and
examined
for
external
abnormalities.
Fetuses
and
placentae
were
pooled
per
litter.
All
samples
were
sent
to
the
Sponsor
(
3M)
for
analysis.
At
this
time,
only
the
liver
and
serum
analyses
have
been
Reported
(
3M
Environmental
Laboratory,
2001).

RESULTS
NOAEL
(
dose
and
effect)
 
maternal
and
developmental:
The
NOAEL
for
maternal
toxicity
is
0.1
mg/
kg/
day.
The
NOAEL
for
developmental
toxicity
is
1.0
mg/
kg/
day.

LOAEL
(
dose
and
effect)
 
maternal
and
developmental:
The
LOAEL
for
maternal
toxicity
is
1.0
mg/
kg/
day,
based
on
abortions,
incidences
of
scant
feces,
and
decreases
in
body
weight
gains
and
food
consumption.
The
LOAEL
for
developmental
toxicity
is
2.5
mg/
kg/
day,
based
on
reductions
in
body
weight
and
increased
incidences
in
fetal
alterations.

Toxic
response/
effects
by
dose
level
­
maternal:
Maternal
toxicity
was
evident
at
dose
levels
of
1.0
mg/
kg/
day
and
above
and
consisted
of
the
following:
abortions
at
2.5
mg/
kg/
day
and
above
occurring
on
GD
22­
28;
increased
incidence
of
scant
feces
at
1.0
mgkgday
and
above;
reductions
in
mean
body
weight
and
body
weight
gain
at
doses
of
1.0
mg/
kg/
day
and
above;
reductions
in
absolute
and
relative
food
consumption
at
2.5
mg/
kg/
day
and
above.

Toxic
response/
effects
by
dose
level
 
developmental:
Developmental
toxicity
was
evident
at
doses
of
2.5
mg/
kg/
day
and
above
and
consisted
of
the
following:
reductions
in
mean
fetal
body
weight
at
2.5
mg/
kg/
day
and
above;
delayed
ossification
at
2.5
mg/
kg/
day
and
above.

Statistical
results:

Maternal
data:
Statistically
significant
increases
in
abortions
were
observed
at
3.75
mg/
kg/
day.
Incidences
of
scant
feces
at
3.75
mg/
kg/
day
reached
statistical
significance
(
p<
0.01).
Maternal
body
weight
gains
were
reduced
or
body
weight
losses
occurred
in
the
1.0,
2.5,
and
3.75
mg/
kg/
day
dosage
groups
at
most
measured
intervals
during
dosing;
these
reductions
were
significant
(
p<
0.05
or
0.01)
in
the
3.75
mg/
kg/
day
dose
group
on
GDs
10­
13,
13­
16,
and
16­
19.
Dosage­
dependent,
significant
body
weight
reductions
or
body
weight
losses
(
p<
0.05
or
0.01)
occurred
in
the
1.0,
2.5,
and
3.75
mg/
kg/
day
dosage
groups
for
the
entire
dosage
period
(
calculated
as
GD
7­
21).
Dosage­
dependent
reductions
in
body
weight
gains
occurred
in
the
2.5
and
3.75
mg/
kg/
day
dosage
groups
for
the
entire
period
of
gestation
(
GD
0­
29)
ENV/
JM/
RD(
2002)
17/
FINAL
314
and
for
the
gestation
period
after
the
initiation
of
dosing
(
GD
7­
29;
significant
at
p<
0.01
in
the
2.5
mg/
kg/
day
dosage
group).
Average
body
weights
were
significantly
reduced
(
p<
0.05
or
0.01)
on
GD
17­
24
in
the
3.75
mg/
kg/
day
dosage
group.
Absolute
and
relative
food
consumption
values
were
significantly
reduced
(
p<
0.05
or
0.01)
in
the
2.5
and
3.75
mg/
kg/
day
dosage
groups
for
the
entire
dosage
period
(
GD
7­
21),
and
the
entire
period
after
the
initiation
of
dosage
(
GD
7­
29).

Fetal
data:
Fetal
body
weights
(
total,
male
and
female)
were
significantly
reduced
(
p<
0.05
and
p<
0.01,
respectively)
in
the
2.75
and
3.75
dosage
groups.
Significant
delays
(
p<
0.05
and
0.01)
in
litter
and
fetal
averages
for
ossification
were
seen
at
both
2.5
and
3.75
mg/
kg/
day
dosage
groups.

Remarks:
Maternal
toxicity
was
evident
at
doses
of
1.0
mg/
kg/
day
and
above.
One
doe
in
the
2.5
mg/
kg/
day
group
and
nine
does
in
the
3.75
mg/
kg/
day
aborted.
All
abortions
occurred
on
gestation
days
22­
28
and
were
considered
treatment­
related
by
the
study
authors.
There
was
a
significant
increase
in
the
incidence
of
scant
feces
in
the
3.75
mg/
kg/
day
group.
Scant
feces
were
also
noted
in
one
and
three
does
in
the
1.0
and
2.5
mg/
kg/
day
groups,
respectively.
Mean
maternal
body
weight
gains
were
significantly
reduced
in
the
3.75
mg/
kg/
day
group
on
gestation
days
10­
13,
13­
16,
16­
19
and
21­
24.
Mean
body
weight
gains
were
also
calculated
for
the
treatment
period
(
days
7­
21),
post­
treatment
period
(
days
21­
29)
and
duration
of
the
study
(
days
7­
29).
There
was
a
significant
reduction
in
mean
maternal
body
weight
gain
during
the
treatment
period
in
the
1.0,
2.5
and
3.75
mg/
kg/
day
groups.
Mean
body
weight
gain
for
the
entire
study
period
was
also
significantly
reduced
in
the
2.5
mg/
kg/
day
group.
Mean
food
consumption
(
g/
kg/
day)
was
significantly
reduced
in
the
2.5
mg/
kg/
day
group
on
gestation
days
16­
19,
19­
21
and
21­
24,
as
well
as
for
the
entire
study
period
(
days
7­
29).
Mean
food
consumption
was
significantly
reduced
in
the
3.75
mg/
kg/
day
group
on
gestation
days
13­
16,
16­
19,
19­
21
and
21­
24,
as
well
as
the
entire
treatment
period
(
GD
7­
21)
and
the
entire
period
after
the
initiation
of
dosage
(
GD
7­
29).
Pregnancy
occurred
in
20
(
90%),
19
(
86.4%),
19
(
86.4%),
17
(
77.3%),
and
21
(
95.4%)
rabbits
in
each
dosage
group.
Ceasarean­
sectioning
observations
on
GD
29
were
based
on
20,
18,
19,
16,
and
12
pregnant
rabbits
in
each
of
the
five
respective
dosage
groups.

In
the
satellite
study
of
does
euthanized
on
gestation
day
21,
the
liver
and
serum
analyses
were
reported
by
3M
Environmental
Laboratory
(
2001).
All
serum
and
liver
samples
(
including
those
from
untreated
controls)
had
detectable
levels
of
PFOS;
the
values
are
presented
below.
Data
from
analysis
of
fetal
and
placenta
tissues
were
not
presented
in
the
report.

Average
Concentration
of
PFOS
in
Rabbit
Liver
and
Serum
by
Dose
Group:
Dose
group
PFOS
conc.
PFOS
conc.
(
mg/
kg/
day)
liver
(
ug/
g)
serum
(
ug/
ml)

0.0
0.239
0.0690
0.1
13.1
2.73
1.0
133
23.8
2.5
317
45.8
3.75
416
88.9
Qualitatively,
increasing
concentrations
of
PFOS
were
found
in
samples
of
liver
and
serum
as
doses
of
PFOS
increased.
The
levels
of
PFOS
are
much
higher
in
the
liver
than
in
the
serum.

These
values
should
be
viewed
with
caution.
It
was
stated
that
because
radio­
labeled
reference
material
was
not
available,
"
it
is
not
possible
to
verify
true
recovery
of
endogenous
analyte
from
tissues."
Matrix
spike
recovery
indicates
the
accuracy
of
quantitation
to
be
+/­
30%.
It
is
also
noted
that
liver
concentrations
may
be
biased
high.
The
only
conclusion
presented
in
the
laboratory
report
is
that
"
PFOS
ENV/
JM/
RD(
2002)
17/
FINAL
315
was
observed
in
the
liver
and
serum
of
all
rabbits
dosed
with
the
test
article."

Developmental
toxicity
was
evident
at
doses
of
2.5
mg/
kg/
day
and
above.
The
number
of
corpora
lutea,
resorptions,
live/
dead
fetuses,
litter
size
and
sex
ratio
were
comparable
among
treated
and
control
groups.
Mean
fetal
body
weight
(
male,
female
and
sexes
combined)
was
significantly
reduced
in
the
2.5
and
3.75
mg/
kg/
day
groups.
There
was
also
a
significant
reduction
in
the
ossification
of
the
sternum
(
litter
averages)
in
the
2.5
and
3.75
mg/
kg/
day
groups,
and
a
significant
reduction
in
the
ossification
of
the
hyoid
(
litter
averages),
metacarpals
(
litter
averages)
and
pubis
(
litter
and
fetal
averages)
in
the
3.75
mg/
kg/
day
group.
Other
fetal
gross
external,
soft
tissue
and
skeletal
alterations
(
malformations
and
variations)
were
considered
unrelated
to
treatment
because
the
incidences
were
not
dosage­
dependent
and/
or
were
within
historical
control
range.

CONCLUSIONS
Conclusions
are
summarized
above
and
this
reviewer
agrees.

REFERENCE
Christian,
M.
S.,
Hoberman,
A.
M.,
and
York,
R.
G.
1999.
Argus
Research
Laboratories,
Inc.
Protocol
Number:
418­
012,
January
1999.
"
Oral
(
Stomach
Tube)
Developmental
Toxicity
Study
of
PFOS
in
Rabbits".

3M
Environmental
Laboratory.
2001.
Analytical
Laboratory
Report,
FACT
TOX­
099,
February
9,
2001.
"
Determination
of
the
Concentration
of
Potassium
Perfluorooctanesulfonate
(
PFOS)
in
Rabbit
Liver
and
Serum
Samples."
ENV/
JM/
RD(
2002)
17/
FINAL
316
REPRODUCTIVE
TOXICITY
STUDIES
Title:
COMBINED
ORAL
(
GAVAGE)
FERTILITY,
DEVELOPMENTAL
AND
PERINATAL/
POSTNATAL
REPRODUCTION
TOXICITY
STUDY
OF
PFOS
IN
RATS
 
ARGUS
RESEARCH
LABORATORIES
STUDY
NUMBER:
6295.9,
1999.

TEST
SUBSTANCE
Identity:
Potassium
Perflurooctylsulfonate,
CAS
No.
2795­
39­
3.

Remarks:
The
test
article,
FC­
95
(
lot
217),
was
received
on
May
20,
1998,
and
stored
at
room
temperature.
Prepared
suspensions
were
stored
at
room
temperature
overnight.
Information
regarding
the
purity,
identity,
strength
and
composition
of
the
test
article
is
on
file
with
the
Sponsor.

METHOD
Method/
Guideline
followed
(
i.
e.,
OECD
414,
etc.):
This
study
was
designed
to
evaluate
ICH
Harmonized
Tripartite
Guideline
stages
A­
F.
A
modification
of
the
requirements
of
the
U.
S.
Food
and
Drug
Administration
(
FDA)
were
used
as
a
basis
for
the
study
design.

Type
of
study
(
one­
generation,
two­
generation,
etc.):
Two­
generation
reproductive
toxicity
GLP
(
Y/
N):
The
study
was
conducted
in
compliance
with
the
Good
Laboratory
Practice
(
GLP)
regulations
of
the
U.
S.
Food
and
Drug
Administration
(
FDA),
the
Japanese
Ministry
of
Health
and
Welfare
(
MHW)
and
the
European
Economic
Community
(
EEC).
There
were
no
significant
deviations
from
the
GLP
regulations
that
affected
the
quality
or
integrity
of
the
study.
Quality
Assurance
Unit
findings
derived
from
the
inspections
during
the
conduct
of
this
study
have
been
documented.

Year
study
performed:
1999
Species/
Strain:
Sprague
Dawley
rats
Sex
(
males/
females/
both):
Both
Number
of
animals
per
dose:
35
Route
of
administration:
Gavage
Dosing
regimen
(
list
all
with
units):
Five
groups
of
35
rats
per
sex
per
dose
group
were
administered
PFOS
by
gavage
for
six
weeks
prior
to
and
during
mating.
Treatment
in
male
rats
continued
until
one
day
before
sacrifice
(
approximately
22
days
total);
female
rats
were
treated
throughout
gestation,
parturition,
and
lactation.

Doses:
0,
0.1,
0.4,
1.6,
and
3.2
mg/
kg/
day
Premating
exposure
period
for
males/
females
(
P
and
F1,
if
appropriate):
Six
weeks
for
P;
Nine
and
a
half
weeks
for
F1.

Statistical
methods
used:
Proportion
data
were
analyzed
using
the
Variance
Test
for
Homogeneity
of
the
Binomial
Distribution.
Continuous
data
(
body
weights,
body
weight
changes,
and
feed
consumption)
were
analyzed
using
Bartlett's
Test
of
Homogeneity
of
Variance
and
Analysis
of
Variance
(
ANOVA).
If
the
ENV/
JM/
RD(
2002)
17/
FINAL
317
ANOVA
was
significant
(
p
<
0.05),
Dunnett's
Test
was
used
to
identify
the
statistical
significance
of
the
individual
groups.
If
the
ANOVA
was
not
appropriate,
the
Kruskal­
Wallis
Test
was
used.
In
cases
where
the
Kruskal­
Wallis
Test
was
statistically
significant
(
p
<
0.05),
Dunn's
Method
of
Multiple
Comparisons
was
used
to
identify
the
statistical
significance
of
the
individual
groups.

If
there
were
greater
than
75%
ties,
Fisher's
Exact
Test
was
used.
Fisher's
Exact
Test
was
also
used
to
evaluate
necropsy
data
for
the
pups
which
were
stillborn
or
found
dead.
Data
obtained
at
Ceasareansectioning
natural
delivery,
preweaning
reflex/
physical
developmental
data
and
postweaning
behavorial
data
involving
discrete
data
(
number
of
corpora
lutea,
number
of
pups
per
litter,
trials
to
a
criterion)
were
evaluated
by
the
Kruskal­
Wallis
Test.

Remarks
 
Detail
and
discuss
any
significant
protocol
parameters
and
deviations:

F0
Generation:

Parental
animals
(
F0)
were
observed
twice
daily
for
clinical
signs.
Body
weights
and
food
consumption
values
were
recorded
weekly
during
the
treatment
period
in
male
rats;
and
weekly
during
mating
and
then
daily
during
gestation,
and
on
lactation
days
1,
4,
7,
10,
14,
and
at
sacrifice
in
female
rats.
Each
dosage
group
consisted
of
two
sets
of
female
rats.
One
set
consisted
of
the
first
ten
female
rats
with
confirmation
of
mating;
this
group
was
dosed
until
gestation
day
(
GD)
10
and
delivered
via
Caesarean­
sectioning.
The
remaining
females
comprised
the
second
set
which
delivered
naturally.
During
the
21­
day
lactation
period,
the
dams
were
evaluated
for
clinical
signs
during
parturition
and
length
of
gestation,
and
then
each
litter
was
evaluated
at
least
twice
daily
for
size
and
pup
viability
at
birth.
Pup
observations
during
the
21­
day
lactation
period
included
physical
signs,
body
weights,
nursing
behavior,
surface
righting
reflex,
pinna
unfolding,
eye
opening,
acoustic
startle
response
and
air
righting
reflex.
Pupil
constriction
was
evaluated
only
on
lactation
day
21.
On
lactation
day
4,
litters
were
randomly
culled
to
four
male
and
four
female
pups.
The
remaining
pups
were
sacrificed
and
necropsied.
The
F0
male
rats
were
sacrificed
and
necropsied
after
the
end
of
dosing
at
the
time
of
parturition
(
lactation
day
1).
The
testes,
epididymides,
prostate,
and
seminal
vesicles
were
weighed.
Evaluations
of
sperm
number,
motility,
and
morphology
were
not
included
in
the
protocol.
The
F0
generation
females
that
delivered
by
Caesarean­
sectioning
were
sacrificed
on
GD
10
and
necropsied.
Pregnancy
status
was
confirmed,
the
ovaries
were
examined
for
the
number
and
distribution
of
corpora
lutea,
implantation
sites
were
determined,
and
embryos
were
examined
for
viability.
The
F0
generation
females
that
delivered
naturally
were
sacrificed
on
lactation
day
(
LD)
21
and
necropsied.
Ovaries
were
examined
as
above
and
the
number
and
distribution
of
implantation
sites
was
recorded.
The
liver
from
each
parental
rat
was
removed,
weighed
and
analyzed.
Blood
samples
were
collected
from
5
male
rats
that
had
mated
and
from
5
female
rats
on
LD
21
for
pharmacokinetic
analysis;
livers
from
the
pups
from
the
litters
of
these
five
dams
were
also
collected
for
analysis.

F1
Generation:

Since
F1
generation
pup
viability
was
significantly
reduced
in
the
1.6
and
3.2
mg/
kg/
day
dosages
groups,
only
the
0.1
and
0.4
mg/
kg/
day
dosage
groups
were
carried
into
the
second
generation.
Twenty­
five
F1
generation
rats
per
sex
per
dose
group
were
administered
PFOS
by
gavage
at
doses
of
0,
0.1,
and
0.4
mg/
kg/
day
beginning
on
LD
22
and
continuing
through
the
day
before
sacrifice.
At
24
days
of
age,
one
rat
per
sex
per
litter
in
each
dosage
group
was
tested
in
a
passive
avoidance
paradigm.
On
LD
28,
females
were
evaluated
for
the
age
of
vaginal
patency
and
on
LD
34,
male
rats
were
evaluated
for
the
age
of
preputial
separation.
One
rat
per
sex
per
litter
were
evaluated
in
a
water­
filled
M­
maze
on
LD
70.
Assignment
to
cohabitation
within
each
dosage
group
began
on
LD
90.
Females
with
evidence
of
mating
were
considered
to
be
at
GD
0
and
assigned
to
individual
housing
for
the
remainder
of
the
dosing
period.
The
F1
generation
male
rats
were
sacrificed
after
mating,
necropsied
and
evaluated
as
described
in
the
F0
generation.
All
F1
generation
females
were
allowed
to
deliver
naturally.
Dams
that
delivered
litters
were
ENV/
JM/
RD(
2002)
17/
FINAL
318
sacrificed
and
necropsied
on
LD
21.
All
F2
generation
pups
were
sacrificed,
necropsied,
and
examined
on
LD
21
as
previously
described
for
the
F1
generation
pups.

RESULTS
NOAEL
(
dose
and
effect)
 
for
F0,
F1,
and
F2
(
as
appropriate):
The
NOAEL
for
the
F0
generation
male
and
female
parental
animals
=
0.1
mg/
kg/
day,
the
lowest
dose
tested.
The
NOAEL
for
the
F1
generation
male
parental
animals
could
not
be
determined
since
treatment
related
signs
of
toxicity
were
observed
at
0.1
and
0.4
mg/
kg/
day;
the
NOAEL
for
the
F1
generation
female
parental
animals
=
0.1
mg/
kg/
day.
The
NOAEL
for
the
F1
generation
offspring
=
0.4
mg/
kg/
day.
The
NOAEL
for
the
F2
generation
offspring
=
0.1
mg/
kg/
day.

LOAEL
(
dose
and
effect)
 
for
F0,
F1,
and
F2
(
as
appropriate):
The
LOAEL
for
the
F0
generation
male
and
female
parental
animals
=
0.4
mg/
kg/
day,
based
on
reductions
in
body
weight
gain
and
food
consumption.
The
LOAEL
for
the
F1
generation
male
animals
=
0.1
mg/
kg/
day
based
on
significant
reductions
in
absolute
food
consumption;
the
NOAEL
for
the
F1
generation
female
animals
=
0.4
mg/
kg/
day
based
on
reductions
in
body
weight
and
food
consumption.
The
LOAEL
for
the
F1
generation
offspring
=
1.6
mg/
kg/
day,
based
on
significant
reductions
in
the
number
of
implantation
sites,
litter
size,
pup
viability,
growth
and
survival.
The
LOAEL
for
the
F2
generation
offspring
=
0.4
mg/
kg/
day,
based
on
significant
reductions
in
pup
growth.

Toxic
response/
effects
by
dose
level
 
parental/
F1:
Toxic
effects
in
F0
generation
animals:
reductions
in
both
body
weight
gains
and
in
absolute
and
relative
food
consumption
at
the
1.6
and
3.2
mg/
kg/
day
dosage
groups
during
the
pre­
mating
period.
Following
mating,
food
consumption
was
significantly
reduced
in
the
0.4.
and
1.6
mg/
kg/
day
dosage
groups.
Terminal
body
weights
were
also
significantly
reduced
in
the
1.6
and
3.2
mg/
kg/
day
dose
groups.
Signs
of
reproductive
toxicity
in
the
F0
generation
males
were
seen
at
the
highest
dose
group
of
3.2
mg/
kg/
day
and
included
significant
reductions
in
the
absolute
weights
of
the
seminal
vesicles
(
with
fluid)
and
the
prostate.
A
significant
increase
in
the
number
of
males
with
brown
liver
at
3.2
mg/
kg/
day
dose
group
was
also
reported.
The
only
findings
reported
in
the
F0
dams
occurred
in
the
0.4,
1.6,
and
3.2
mg/
kg/
day
dosage
groups
and
included
localized
alopecia
during
pre­
mating,
gestation,
and
lactation;
and
reductions
in
body
weight
and
food
consumption
values
observed
during
the
pre­
mating
period
and
continuing
throughout
gestation
and
lactation.

Toxic
effects
in
F1
generation
animals:
F1
males;
the
only
reported
effects
were
significant
reductions
in
absolute
food
consumption
on
postweaning
days
1­
8
occurring
at
the
0.1
and
0.4
mg/
kg/
day
dose
levels.
F1
females;
observations
at
the
0.4
mg/
kg/
day
dosage
group
included,
reductions
in
body
weights
on
day
1
postweaning,
significant
losses
in
body
weight
on
LDs
1­
4,
and
significant
reductions
in
food
consumption
on
days
1­
8
postweaning
and
during
lactation.

Toxic
response/
effects
by
dose
level
 
offspring
(
F1/
F2):

Toxic
effects
in
the
F1
generation
pups
consisted
of
reduced
pup
viability
at
the
two
highest
dosegroups
(
1.6
and
3.2
mg/
kg/
day).
The
reductions
in
pup
viability
began
to
appear
on
LD
4
postculling
in
the
1.6
mg/
kg/
day
dose
group,
with
over
26%
of
the
pups
found
dead
between
LD
2­
4.
In
the
3.2
mg/
kg/
day
dose
group
45%
of
the
pups
were
found
dead
on
LD1;
no
pups
survived
beyond
LD
1.
Statistically
significant
increases
were
observed
in
the
number
of
dams
with
stillborn
pups
at
the
3.2
mg/
kg/
day
dose
group.
Viability
and
lactation
indices
were
significantly
reduced
in
these
same
dosage
groups
(
viability
index
=
0%
at
3.2
mg/
kg/
day
and
66%
at
1.6
mg/
kg/
day;
lactation
index
=
94.6%
at
1.6
mg/
kg/
day).
Statistically
significant
reductions
in
pup
body
weights
were
also
observed
at
the
two
highest
dosage
groups.
Gestation
ENV/
JM/
RD(
2002)
17/
FINAL
319
length
was
significantly
reduced
at
3.2
mg/
kg/
day.
Significant
reduction
in
the
number
of
implantation
sites
followed
by
a
concomitant
reduction
in
litter
size
was
observed
at
3.2
mg/
kg/
say.
Other
adverse
signs
in
the
3.2
mg/
kg/
day
dose
level
associated
with
reductions
in
pup
viability
and
maternal
care
included
litters
with
pups
that
were
not
nursing
or
who
had
no
evidence
of
milk
in
the
stomach,
as
well
as
maternal
cannibalization
of
pups
that
were
stillborn
or
found
dead.

Toxic
effects
in
the
F2
generation
pups
consisted
of
reductions
in
mean
pup
body
weights
(
on
a
per
litter
basis)
observed
at
0.1
mg/
kg/
day
on
LD
4
and
7.
At
0.4
mg/
kg/
day,
statistically
significant
reductions
in
mean
pup
body
weights
were
observed
on
LDs
7­
14.

Statistical
results:
F0
generation
male
animals:
Significant
reductions
(
p
<
0.05
or
p<
0.01)
in
body
weight
gains
at
0.4
mg/
kg/
day
and
higher.
Absolute
and
relative
food
consumption
values
were
significantly
reduced
(
p
<
0.05
or
p
<
0.01)
in
the
1.6
and
3.2
mg/
kg/
day
dosage
groups.
A
significant
increase
(
p
<
0.01)
in
the
number
of
male
rats
in
the
3.2
mg/
kg/
day
dosage
group
with
brown
liver.
The
gross
lesions
of
the
liver
were
considered
to
be
treatment
related
because
the
incidences
were
dosage­
dependent.
Significant
reductions
(
p
<
0.05
and
p
<
0.01)
in
terminal
body
weights
were
observed
in
the
1.6
and
3.2
mg/
kg/
day
dosage
groups.
Significant
reductions
(
p
<
0.05
or
p
<
0.01)
in
the
absolute
weights
of
the
seminal
vesicles
with
fluid
and
the
prostate
were
observed
in
the
3.2
mg/
kg/
day
dosage
group.
F0
generation
female
animals:
Significant
increases
(
p
<
0.05
or
p
<
0.01)
in
localized
alopecia
were
observed
in
the
0.4,
1.6,
and
3.2
mg/
kg/
day
dosages
groups.
Significant
reductions
in
body
weight
and
body
weight
gains
and
food
consumption
(
p
<
0.05
or
p
<
0.01)
were
observed
in
the
1.6
and
3.2
mg/
kg/
day
dosage
during
premating
and
gestation
and
then
in
0.4
mg/
kg/
day
dosage
group
and
above
during
lactation.
Significant
reductions
(
p
<
0.01)
in
gestation
length,
implantation
sites,
and
litter
size
were
observed
at
3.2
mg/
kg/
day.
F1
generation
offspring:
Pup
viability
was
significantly
reduced
(
p
<
0.05
or
p
<
0.01)
in
the
1.6
and
3.2
mg/
kg/
day
dosage
groups.
Significant
increases
(
p
<
0.05
or
p
<
0.01)
were
observed
in
the
number
of
dams
with
stillborn
pups,
while
significant
reductions
(
p
<
0.05
or
p
<
0.01)
were
observed
in
the
viability
index,
lactation
index,
and
averages
for
surviving
pups
at
3.2
mg/
kg/
day.
A
dosage­
dependent
pattern
of
reduced
pup
body
weight
was
evident
in
each
dosage
group,
with
statistical
significance
(
p
<
0.01)
in
the
1.6
and
3.2
mg/
kg/
day
dosage
groups.
F1
generation
adult
animals:
Males
­
Significant
reductions
(
p
<
0.05
or
p
<
0.01)
in
absolute
food
consumption
at
0.1
and
0.4
mg/
kg/
day;
females
­
significant
(
p
<
0.05)
body
weight
loss
on
lactation
days
1­
4
at
0.4
mg/
kg/
day;
and
significant
reductions
(
p
<
0.05)
in
food
consumption
at
0.4
mg/
kg/
day
on
days
1­
8
postweaning.
F2
generation
offspring:
Pup
body
weights
tended
to
be
reduced
in
the
0.1
mg/
kg/
day
dosage
group
on
lactation
day
4
and
7,
but
were
comparable
to
controls
by
lactation
day
14.
Pup
body
weights
in
the
0.4
mg/
kg/
day
dosage
group
tended
to
be
reduced,
though
not
significantly,
on
lactation
days
4­
21,
with
significant
reductions
(
p
<
0.05
and
p
<
0.01,
respectively)
on
lactation
days
7
and
14,
as
compared
to
controls.

Remarks:
In
the
F0
generation
male
rats,
there
were
no
treatment­
related
clinical
signs
of
toxicity,
no
mortality,
and
no
effects
on
mating
or
on
any
of
the
fertility
parameters
evaluated
in
any
dose
group
tested.
Reported
effects
included
reductions
in
both
body
weight
gains
and
in
absolute
and
relative
food
consumption
at
the
1.6
and
3.2
mg/
kg/
day
dosage
groups
during
the
pre­
mating
period.
Following
mating,
food
consumption
was
significantly
reduced
in
the
0.4
and
1.6
mg/
kg/
day
dosage
groups.
Terminal
body
weights
were
also
significantly
reduced
in
the
1.6
and
3.2
mg/
kg/
day
dose
groups.
Body
weights,
body
weight
gains,
absolute
and
relative
food
consumption
were
unaffected
by
treatment
at
the
0.1
mg/
kg/
day
dosage
group.
Signs
of
reproductive
toxicity
in
the
F0
generation
males
were
seen
at
the
highest
dose
group
of
3.2
mg/
kg/
day
and
included
significant
reductions
in
the
absolute
weights
of
the
seminal
vesicles
ENV/
JM/
RD(
2002)
17/
FINAL
320
(
with
fluid)
and
the
prostate.
A
significant
increase
in
the
number
of
males
with
brown
liver
at
3.2
mg/
kg/
day
dose
group
was
also
reported.

In
the
F0
generation
female
rats,
no
deaths
were
reported
at
any
dose
level.
In
dams
sacrificed
on
GD
10
for
Caesarean­
sectioning,
there
did
not
appear
to
be
any
effects
on
estrous
cycling,
mating
and
fertility
parameters,
the
numbers
of
corpora
lutea
and
implantations,
or
in
the
number
of
viable
or
non­
viable
embryos.
The
only
findings
reported
in
the
F0
dams
occurred
in
the
0.4,
1.6,
and
3.2
mg/
kg/
day
dosage
groups
and
included
localized
alopecia
during
pre­
mating,
gestation,
and
lactation;
and
reductions
in
body
weight
and
body
weight
gain
and
food
consumption
values
observed
during
the
pre­
mating
period
and
continuing
throughout
gestation
and
lactation.

Reversible
delays
in
reflex
and
physical
development
were
observed
in
the
F1
generation
offspring.
The
ability
to
surface
right
was
significantly
delayed
in
the
1.6
and
3.2
mg/
kg/
day
dosage
groups
on
LDs
3­
10
(
delays
in
the
3.2
mg/
kg/
day
dose
group
were
observed
on
LD
1,
after
which
there
were
no
surviving
pups
remaining
for
further
observation).
By
the
end
of
the
observation
period,
however,
all
surviving
pups
in
the
1.6
mg/
kg/
day
dosage
group
had
the
ability
to
surface
right.
There
were
no
delays
observed
in
the
ability
to
surface
right
in
dose
groups
<
0.4
mg/
kg/
day.
Similar
responses
were
seen
for
pinna
unfolding
and
eye
opening.
Although
there
were
transient
delays
seen
with
these
signs
of
physical
development
across
all
dose
groups,
by
the
end
of
the
observation
period
responses
in
pups
were
similar
to
controls.
The
time
of
development
of
the
acoustic
startle
reflex
and
the
ability
to
air
right
were
both
significantly
reduced
in
the
1.6
mg/
kg/
day
dosage
group.
No
effects
on
these
reflexes
were
observed
in
the
low
dose
group
of
0.1
mg/
kg/
day
and
only
a
transient
delay
(
on
LD
16
only)
in
the
ability
to
air
right
was
seen
in
the
0.4
mg/
kg/
day
group.
At
the
end
of
lactation
(
LD
21),
all
live
pups
in
all
dose
groups
(
0,
0.1,
0.4,
and
1.6
mg/
kg/
day)
had
pupil
constriction
response.

The
most
significant
finding
reported
in
the
offspring
was
that
of
reduced
pup
viability
at
the
two
highest
dose
groups.
The
reductions
in
pup
viability
began
to
appear
on
LD
4
postculling
in
the
1.6
mg/
kg/
day
dose
group,
with
over
26%
of
the
pups
found
dead
between
LD
2­
4.
In
the
3.2
mg/
kg/
day
dose
group
45%
of
the
pups
were
found
dead
on
LD1;
no
pups
survived
beyond
LD
1.
As
a
result,
the
viability
index
was
greatly
reduced
in
these
dosage
groups
(
0%
at
3.2
mg/
kg/
day
and
66%
at
1.6
mg/
kg/
day).
The
lactation
index
was
also
significantly
reduced
(
94.6%)
in
the
1.6
dosage
group.
In
addition,
gestation
length
was
significantly
reduced
in
the
high­
dose
group
and
there
also
was
a
significant
reduction
in
the
number
of
implantation
sites
followed
by
a
concomitant
reduction
in
litter
size.
Statistically
significant
reductions
in
pup
body
weights
were
also
observed
at
the
two
highest
dosage
groups.
Other
adverse
signs
in
the
3.2
mg/
kg/
day
dose
level
associated
with
reductions
in
pup
viability
and
maternal
care
included
litters
with
pups
that
were
not
nursing
or
who
had
no
evidence
of
milk
in
the
stomach,
as
well
as
maternal
cannibalization
of
pups
that
were
stillborn
or
found
dead.
The
percentage
of
male
pups
was
comparable
across
all
dosage
groups.

Since
F1
generation
pup
viability
was
significantly
reduced
in
the
1.6
and
3.2
mg/
kg/
day
dosages
groups,
only
the
0.1
and
0.4
mg/
kg/
day
dosage
groups
were
carried
into
the
second
generation.

Clinical
observations
in
the
F1
generation
male
rats
appeared
unremarkable.
No
treatment­
related
deaths
were
reported
and
no
statistically
significant
differences
were
reported
for
any
of
the
following
parameters:
body
weights/
body
weight
gains,
average
day
of
preputial
separation;
values
for
learning,
short­
term
retention,
long­
term
retention
or
response
inhibition
as
evaluated
by
performance
in
a
passive
avoidance
or
watermaze
performance
paradigm;
mating
or
fertility
parameters;
necroscopic
examinations;
absolute
or
relative
weights
for
the
right
or
left
testis,
seminal
vesicles,
right
epididymis,
or
prostate;
and
terminal
body
weights.
The
only
reported
effects
were
significant
reductions
in
absolute
food
consumption
on
postweaning
days
1­
8
occurring
at
the
0.1
and
0.4
mg/
kg/
day
dose
levels.
ENV/
JM/
RD(
2002)
17/
FINAL
321
Clinical
observations
for
the
F1
generation
females
were
likewise
unremarkable.
Observations
at
the
0.4
mg/
kg/
day
dosage
group
included,
reductions
in
body
weights
on
day
1
postweaning,
significant
losses
in
body
weight
on
LDs
1­
4,
and
significant
reductions
in
food
consumption
on
days
1­
8
postweaning
and
during
lactation.
There
were
no
statistically
significant
differences
reported
for
any
of
the
following
parameters:
values
for
learning,
short­
term
retention,
long­
term
retention
or
response
inhibition
as
evaluated
by
performance
in
a
passive
avoidance
or
watermaze
performance
paradigm;
mating
and
fertility
parameters;
gestation
index;
pregnancy
rates;
and
necroscopic
examinations.

Evidence
of
treatment­
related
effects
in
the
F2
generation
pups
consisted
of
reductions
in
mean
pup
body
weights
(
on
a
per
litter
basis)
observed
at
0.1
mg/
kg/
day
on
LD
4
and
7.
Body
weights
were
comparable
to
control
levels
by
LD
14.
At
0.4
mg/
kg/
day,
statistically
significant
reductions
in
mean
pup
body
weights
were
observed
on
LDs
7­
14.
Mean
body
weights
on
LD21
continued
to
remain
lower
than
controls,
although
the
difference
was
not
statistically
significant
(
46.5
g
in
0.4
mg/
kg/
day
dose
group
vs.
50
g
in
controls).
Clinical
and
necroscopic
observationsof
the
F2
generation
pups
were
unremarkable.
No
other
toxicologically
significant
effects
were
reported.

Liver
and
sera
samples
collected
from
the
initial
population
of
dosed
animals
(
F0)
and
their
offspring
(
F1)
were
analyzed
for
the
presence
of
PFOS.
The
F0
results
for
the
F0
animals
were:

Dose
group
Average
PFOS
conc.
Average
PFOS
conc.
(
mg/
kg/
day)
in
serum
(
ug/
ml)
in
liver
(
ug/
g)

0.0:
female
0.0307
female
0.171
male
0.0244
male
0.665
0.1:
female
5.28
female
14.8
male
10.5
male
84.9
0.4:
female
18.9
female
58.0
male
45.4
male
176
1.6:
female
82
female
184
male
152
male
323
3.2:
female
NR*
female
NR*
male
273
male
1360
*
samples
not
received
Qualitatively,
the
F0
results
indicate
all
rats
(
including
controls)
had
detectable
levels
of
PFOS
in
serum
and
livers.
PFOS
concentration
increased
with
dose.
PFOS
concentrations
were
higher
in
the
liver
than
in
the
serum,
and
males
had
greatly
increased
PFOS
concentrations
in
serum
and
liver
when
compared
with
females
of
the
same
dose
group.
Pooled
liver
samples
from
the
F1animals
sacrificed
shortly
after
birth
had
lower
PFOS
concentrations
than
adults
of
the
F0
generation
of
the
same
dose
group.
The
average
PFOS
concentrations
in
pooled
liver
samples
from
F1
animals
shortly
after
birth
were
0.0511,
6.19,
57.6,
and
70.4
ug/
g
in
the
0.0,
0.1,
0.4,
and
1.6
mg/
kg/
day
dose
groups,
respectively.
These
quantitative
values
for
the
PFOS
concentration
in
the
liver
and
serum
should
be
viewed
with
caution.
The
accuracy
of
quantitation
is
+
30%,
the
purity
of
the
analytical
reference
substance
is
unknown,
and
there
were
several
uncorrected
dilution
errors.

CONCLUSIONS
Conclusions
stated
above
and
this
reviewer
agrees.
ENV/
JM/
RD(
2002)
17/
FINAL
322
REFERENCE
Christian,
M.
S.,
Hoberman,
A.
M.,
and
York,
R.
G.
1999b.
Argus
Research
Laboratories,
Inc.
Protocol
Number:
418­
008,
Sponsor
Study
Number:
6295.9,
June
10,
1999.
Combined
Oral
(
Gavage)
Fertility,
Developmental
and
Perinatal/
Postnatal
Reproduction
Toxicity
Study
of
PFOS
in
Rats.

3M
Environmental
Laboratory.
1999b.
Analytical
Laboratory
Report
on
the
Determination
of
the
Presence
and
Concentration
of
Potassium
Perfluorooctanesulfonate
(
CAS
Number:
2795­
39­
3)
in
the
Serum
and
Liver
of
Sprague­
Dawley
Rats
Exposed
to
PFOS
via
Gavage.
As
amended
April
19,
2000.
ENV/
JM/
RD(
2002)
17/
FINAL
323
CROSS­
FOSTERING/
DEVELOPMENTAL
TOXICITY
STUDY
Title:
ORAL
(
GAVAGE)
CROSS­
FOSTERING
STUDY
OF
PFOS
IN
RATS
 
ARGUS
RESEARCH
LABORATORIES
STUDY
NUMBER
418­
014,
3M
T­
6295.13,
JULY
1999.

TEST
SUBSTANCE
Identity:
Potassium
Perfluorooctylsulfonate,
CAS
No.
2795­
39­
3.

Remarks:
The
test
article,
FC­
95
(
Lot
217)
was
received
on
October
21,
1998,
and
stored
at
room
temperature.
Prepared
suspensions
were
stored
at
room
temperature.
Information
regarding
the
identity,
strength,
composition,
and
purity
of
the
test
article
is
on
file
with
the
sponsor.

METHOD
Method/
Guideline
followed
(
i.
e.,
OECD
414,
etc.):
For
the
cross­
fostering
study:
The
requirements
of
the
U.
S.
Food
and
Drug
Administration
(
FDA)
were
used
as
the
basis
of
the
study
design,
i.
e.,
the
International
Conference
on
Harmonization:
Guideline
on
detection
of
toxicity
to
reproduction
for
medicinal
products.
Federal
Register,
September,
22,
1994,
Vol.
59.
No.
183.
For
the
PFOS
bioanalysis:
US
FDA
GLP
Final
Rule
21
CFR
58,
with
exceptions:
Two
separate
study
directors
were
assigned
to
the
in­
life
phase
and
the
analytical
phase
of
this
study.
The
ABS
final
report
does
not
have
a
Statement
of
Compliance.
The
QAU
statement
in
the
ABS
final
report
indicates
compliance
with
EPA
40
CFR
Part
792,
rather
than
FDA
21
CFR
Part
58.
Dose
confirmation
analyses
were
not
conducted
according
to
the
GLP
regulations;
analytical
method
was
not
fully
validated.
Not
all
raw
data
were
verified
by
the
group
leader
or
designee.

GLP
(
Y/
N):
The
study
was
conducted
in
compliance
with
the
Good
Laboratory
Practice
(
GLP)
regulations
of
the
U.
S.
FDA,
the
Japanese
Ministry
of
Health
and
Welfare,
and
the
European
Economic
Community.
There
were
no
deviations
from
the
GLP
regulations
that
affected
the
quality
or
integrity
of
the
study.
Quality
assurance
Unit
findings
derived
from
the
inspections
during
the
conduct
of
this
study
are
documented.

Year
study
performed:
1999
Species/
Strain:
Sprague­
Dawley
rats
Number
of
animals
per
dose:
25
Route
of
administration:
Gavage
Dosing
regimen
(
list
all
with
units):
Two
groups
of
25
female
rats
were
administered
PFOS
by
gavage
beginning
42
days
prior
to
mating
to
untreated
(
breeder)
males,
and
continuing
throughout
gestation
and
into
day
21
of
lactation.

Doses:
0,
1.6
mg/
kg/
day
Statistical
methods
used:
Averages
and
percentages
were
calculated.
Litter
values
were
used
where
appropriate.

Remarks
 
Detail
and
discuss
any
significant
protocol
parameters
and
deviations:
Two
groups
of
25
ENV/
JM/
RD(
2002)
17/
FINAL
324
female
Sprague­
Dawley
rats
were
administered
0
and
1.6
mg/
kg/
day
PFOS
in
0.5%
Tween­
80
by
gavage,
beginning
42
days
prior
to
mating
to
untreated
(
breeder)
males,
and
continuing
throughout
gestation
and
into
day
21
of
lactation.
A
dose
volume
of
5
mL/
kg
was
administered,
adjusted
daily
on
the
basis
of
individual
body
weight.
Parental
females
were
observed
twice
daily
for
viability
and
clinical
observations
were
recorded
1
hour
prior
to
and
after
dosing
during
the
treatment
period.
Maternal
body
weights
were
recorded
once
during
the
acclimation
period
and
then
daily
during
the
treatment
period
and
at
sacrifice;
food
consumption
was
also
recorded
once
during
the
acclimation
period
and
then
daily
during
gestation
and
on
days
1,
4,
7,
10,
and
14
of
lactation.
During
parturition,
females
were
continually
evaluated
for
clinical
signs
and
also
for
duration
of
gestation,
length
of
parturition,
litter
sizes,
and
pup
viability
at
birth.
Maternal
behavior
was
recorded
daily
throughout
lactation.
All
maternal
rats
were
sacrificed
by
carbon
dioxide
asphyxiation
on
day
22
of
lactation
and
a
gross
necropsy
of
the
thoracic,
abdominal,
and
pelvic
viscera
was
performed;
any
gross
lesions
were
preserved
for
future
analysis.
In
addition,
the
number
and
distribution
of
implantation
sites
were
recorded.
Rats
that
did
not
deliver
a
litter
were
not
included
in
the
cross­
fostering
procedure
and
were
sacrificed
on
lactation
day
25,
examined
for
gross
lesions,
and
the
uteri
examined
to
confirm
the
presence/
absence
of
implantation
sites.
Dams
with
no
surviving
pups
were
sacrificed
after
the
last
pup
was
found
dead,
missing,
or
presumed
cannibalized.

Following
completion
of
parturition,
litters
were
immediately
removed
from
their
respective
dams
and
placed
with
either
a
control­
or
PFOS­
treated
dam
for
rearing.
This
cross­
fostering
procedure
resulted
in
four
groups
of
12­
13
dams
or
pups
as
follows:
A)
control
dams
with
litters
from
PFOS­
treated
dams,
i.
e.,
in
utero
exposure
only;
B)
control
dams
with
litters
from
control
dams,
i.
e.,
negative
control;
C)
PFOStreated
dams
with
litters
from
PFOS­
treated
dams,
i.
e.,
both
in
utero
and
post­
natal
exposure;
and
D)
PFOS­
treated
dams
with
litters
from
control
dams,
i.
e.,
post­
natal
exposure
only.

On
day
1
of
lactation
(
birth),
each
pup
was
individually
weighed
and
each
litter
was
evaluated
twice
daily
during
lactation
for
viability.
Pups
were
observed
once
daily
for
clinical
signs
and
gross
external
physical
anomalies.
Pup
body
weights
were
recorded
on
days
1,
4,
7,
14,
and
21of
lactation,
and
then
at
sacrifice.
On
day
4
of
lactation,
each
cross­
fostered
litter
was
culled
to
5
males
and
5
females.
On
day
21
of
lactation,
all
pups
were
sacrificed
via
decapitation
and
examined
for
gross
lesions.
Pups
found
dead
or
sacrificed
because
of
moribundity
were
examined
for
gross
lesions
and
for
the
cause
of
death
or
the
moribund
condition.
The
lungs,
liver,
and
any
gross
lesions
were
collected
from
selected
pups
at
various
timepoints
(
for
the
F0
generation,
blood,
milk,
and
liver
samples
were
collected
on
LD
14;
milk
was
collected
2­
6
hours
post­
dosing;
for
the
F1
generation
offspring,
blood
was
collected
on
LD
14)
and
preserved
for
possible
future
analysis.
Liver
samples
were
evaluated
via
electron
microscopy.
In
addition,
samples
of
blood,
milk
(
including
the
milk­
secreting
glands),
and
liver
were
collected
from
selected
maternal
rats
and
pups
(
blood
and
liver
samples
were
pooled
per
litter)
at
various
timepoints
for
analysis
of
PFOS
concentration.
Only
the
analysis
of
the
sera
samples
are
available
at
this
time.

RESULTS
Toxic
response/
effects
by
dose
level
­
maternal:
Mean
maternal
body
weight
and
body
weight
gains
at
1.6
mg/
kg/
day
were
reduced
compared
to
controls
during
premating
and
continuing
throughout
gestation.
Mean
maternal
body
weight
changes
in
the
treated
group
were
comparable
to
those
seen
in
control
animals
during
lactation.
During
the
premating
period,
and
on
into
gestation
and
lactation,
food
consumption
was
reduced
in
treated
animals
as
compared
to
controls.
Reductions
in
gestation
length,
the
average
number
of
implantation
sites,
delivered
sizes,
and
live
litter
size
were
observed
in
treated
animals.

Toxic
response/
effects
by
dose
level
 
developmental:
Pup
mortality
was
observed
in
two
of
the
crossfostered
groups.
On
lactation
days
2­
4,
approximately
19%
of
the
pups
in
group
C
were
either
found
dead
or
presumed
cannibalized.
Pup
mortality
was
also
observed
in
group
A
at
a
rate
of
9%.
In
addition,
the
ENV/
JM/
RD(
2002)
17/
FINAL
325
number
of
live
pups
on
day
4
of
lactation,
numbers
of
surviving
pups
per
litter,
and
live
litter
sizes
were
also
reduced
in
these
two
groups.
Pup
mortality
in
groups
B
and
D
during
lactation
days
2­
4
were
at
1.6%
and
1.1%,
respectively.
Reductions
in
pup
body
weights
were
observed
in
groups
A
and
C
on
day
1
of
lactation.
Pup
body
weights
in
group
D
were
comparable
to
controls
during
that
same
period.
From
lactation
day
4­
21,
pup
body
weights
in
groups
A,
C,
and
D
were
reduced
when
compared
to
group
B
(
negative
control),
with
the
reductions
greatest
in
group
C.
Two
litters
in
group
A
and
one
litter
on
group
C
did
not
nurse.
Milk
analysis
of
the
stomachs
of
pups
found
no
milk
in
the
stomachs
of
57%,
100%,
and
87%
of
the
pups
found
dead
and
necropsied
in
groups
A,
C,
and
D,
respectively.

Electron
microscopic
examination
of
the
liver
revealed
an
increase
in
the
number
of
peroxisomes
in
pups
from
dams
treated
with
1.6
mg/
kg/
day
PFOS.

Statistical
results:
The
method
of
statistical
analysis
consisted
of
calculation
of
averages
(
mean
+
S.
D.)
and
percentages.
Statistical
significance
was
not
assessed.
The
data
were
presented
according
to
day
of
gestation/
lactation
and
compared
by
dose
and
by
cross­
fostered
group.
Litter
values
were
used
where
appropriate.
The
data
are
too
cumbersome
to
cite
here.

Remarks
 
Additional
information
to
adequately
assess
the
data:
All
maternal
rats
survived
to
schedule
sacrifice.
Signs
of
clinical
toxicity
observed
in
the
dams
during
the
study
period
(
e.
g.,
chromorhinorrhea,
scaly
tail,
abrasion
on
the
head,
neck,
tail
and/
or
forelimb,
missing,
broken
and/
or
misaligned
incisors,
and
localized
alopecia,
among
others)
were
not
considered
to
be
treatment­
related
since
they
also
occurred
in
the
control
animals.
All
pregnant
animals
delivered
live
offspring.

Following
cross­
fostering
on
LD
1,
live
litter
sizes
were
comparable
between
treated
and
control.
Sex
ratios
and
the
lactation
index
were
comparable
among
all
groups.
Signs
of
clinical
toxicity
were
observed
in
pups,
but
were
not
considered
to
be
treatment­
related
since
they
also
occurred
in
group
B
(
negative
control)
at
the
same
rate.
No
significant
differences
were
observed
between
group
B
and
the
other
groups
following
examination
of
pup
lungs.

PFOS
concentrations
in
the
serum
of
untreated
dams
ranged
from
below
the
limit
of
detection
(
0.05
ug/
ml)
to
5.34
ug/
ml.
Serum
PFOS
concentrations
in
the
pups
from
untreated
dams,
fostered
with
untreated
dams,
were
below
the
limit
of
detection.
Serum
PFOS
concentrations
in
the
pups
from
treated
dams,
fostered
with
untreated
dams,
ranged
from
47.6
ug/
ml
to
59.2
ug/
ml.
PFOS
concentrations
in
the
serum
of
treated
dams
ranged
from
59.2
ug/
ml
to
157
ug/
ml.
Serum
PFOS
concentrations
in
the
pups
from
untreated
dams,
fostered
with
treated
dams,
ranged
from
below
the
limit
of
detection
to
35.7
ug/
ml.
Serum
PFOS
concentrations
in
the
pups
from
treated
dams,
fostered
with
treated
dams,
ranged
from
79.5
ug/
ml
to
96.9
ug/
ml.
These
data
indicate
that
exposure
to
PFOS
can
occur
both
in
utero
and
via
milk
from
treated
dams.

CONCLUSIONS
Pups
from
control
dams
that
were
cross­
fostered
with
PFOS­
treated
dams
(
post­
natal
exposure
only)
had
the
same
low
mortality
rate
(
1.1%)
as
pups
from
control
dams
cross­
fostered
with
control
dams
(
1.6%;
negative
control).
Mortality
rates
in
the
remaining
two
groups,
however
(
i.
e.
control
dams
with
litters
from
PFOS­
treated
dams,
i.
e.,
in
utero
exposure
only;
and
PFOS­
treated
dams
with
litters
from
PFOS­
treated
dams,
i.
e.,
both
in
utero
and
post­
natal
exposure),
had
higher
mortality
rates
at
9.6
%
and
19.2%,
respectively.
Under
the
conditions
of
the
study,
this
limited
data
appear
to
indicate
that
reduced
pup
survival
is
mainly
a
result
of
in
utero
exposure
to
PFOS
and
that
post­
natal
exposure
via
milk
in
conjunction
with
in
utero
exposure
may
also
contribute
to
reduced
pup
survival.
In
contrast,
exposure
during
lactation
alone,
through
milk
from
exposed
dams,
does
not
appear
to
have
any
adverse
affect
on
pup
viability.
Additionally,
analysis
of
PFOS
concentration
showed
that
PFOS
was
observed
in
the
sera
of
F0
female
rats
exposed
during
the
in­
life
phase
of
the
study.
Additionally,
PFOS
was
observed
in
sera
ENV/
JM/
RD(
2002)
17/
FINAL
326
samples
taken
from
F1
generation
pups
from
female
rats
exposed
to
the
test
substance,
and
in
F1
generation
pups
exposed
via
lactation,
but
not
exposed
in
utero.
These
are
the
conclusions
of
the
study
authors
and
this
reviewer
agrees.

REFERENCE
Christian,
M.
S.,
Hoberman,
A.
M.,
and
York,
R.
G.
1999c.
Testing
Facility:
Argus
Research
Laboratories,
Inc.,
Protocol
Number
418­
014,
Sponsor:
3M
Corporate
Toxicology,
Study
Number
T­
6295.13,
July
23,
1999.
Oral
(
Gavage)
Cross­
Fostering
Study
of
PFOS
in
Rats.

3M
Environmental
Laboratory.
1999.
Analytical
Laboratory
Report
on
the
Determination
of
the
Presence
and
Concentration
of
Perfluorooctanesulfonate
(
PFOS)
(
CAS
Number:
2759­
39­
3)
in
the
Serum
of
Sprague­
Dawley
Rats
Exposed
to
Potassium
Perfluorooctanesulfonate
via
Gavage,
Laboratory
Report
No.
U2779,
Requestor
Project
No.
3M
Tox
6295.13.
Study
initiation
date:
June
10,
1999.
Completion
at
signing
(
2/
11/
00).
Sample
analysis
completion
June
28,
1999.
Study
Identification
Number:
FACT
Tox­
108
ENV/
JM/
RD(
2002)
17/
FINAL
327
REPEAT
DOSE
DATA
Title:
104­
Week
Dietary
Chronic
Toxicity
and
Carcinogenicity
Study
with
Perfluorooctane
Sulfonic
Acid
Potassium
Salt
(
PFOS;
T­
6295)
in
Rats
TEST
SUBSTANCE
Identity:
Perfluorooctane
Sulfonic
Acid
Potassium
Salt
(
PFOS;
T­
6295).

Remarks:
The
test
substance,
Lot
No.
217,
is
a
light
colored,
free
flowing
powder,
and
is
86.9%
pure.
The
impurities
of
the
test
substance
were
not
indicated
in
the
main
body
of
the
study
report.

METHOD
Method/
guideline
followed:
Guideline
number
not
stated
Study
duration:
Two
years
GLP
(
Y/
N):
Yes
Year
study
performed:
1998­
2000
Species/
strain:
Sprague­
Dawley
rat
[
Crl:
COBSR
CD(
SD)
BR]

Sex:
Male
and
female
Number
of
animals
per
dose
group:
The
control
and
high­
dose
groups
contained
70
rats/
sex
and
the
low­,
mid­,
and
mid­
high
dose
groups
contained
60
rats/
sex.
A
high­
dose
recovery
group
contained
40
rats/
sex.

Route
of
administration:
Diet
Doses
tested
and
frequency:
Control
(
Group
1):
0
ppm
;
low­
dose
(
Group
2):
0.5
ppm
;
mid
dose
(
Group
3):
2
ppm;
mid­
high
dose
(
Group
4):
5
ppm;
high­
dose
(
Group
5)
:
20
ppm.

Post­
treatment
observation
period:
A
high­
dose
recovery
group
(
Group
6)
was
observed
for
52
weeks
after
52
weeks
of
treatment.

Statistical
methods
used:
Levene's
test
was
done
to
test
for
variance
homogeneity.
In
the
case
of
heterogeneity
of
variance
at
P<
0.05,
transformations
were
used
to
stabilize
the
variance.
One­
way
analysis
of
variance
[
ANOVA]
was
used
to
analyze
body
weights,
body
weight
changes,
food
consumption,
continuous
clinical
pathology
values,
palmitoyl
CoA
oxidase
activities,
and
organ
weight
data.
ANOVA
was
done
on
the
homogeneous
or
transformed
data.
If
the
ANOVA
was
significant,
Dunnett's
t­
test
was
used
for
pairwise
comparisons
between
treated
and
control
groups.
Group
comparisions
were
evaluated
at
the
5.0%,
two­
tailed
probability
level.

Remarks:
The
animals
were
observed
twice
daily
(
a.
m.
and
p.
m.)
for
mortality
and
moribundity;
findings
were
recorded
as
they
were
observed.
At
least
once
prior
to
treatment
and
weekly
thereafter,
each
animal
was
removed
from
its
cage
and
examined;
abnormal
findings
or
an
indication
of
normal
was
recorded.
Body
weight
data
were
collected
weekly
through
Week
17,
once
every
4
weeks
thereafter,
and
at
Week
ENV/
JM/
RD(
2002)
17/
FINAL
328
105.
Food
consumption
data
were
collected
weekly
for
the
first
16
weeks
and
once
every
4
weeks
thereafter.
During
Weeks
4,
14,
27,
and
53,
blood
and
urine
were
collected
for
hematology,
clinical
chemistry,
urinalysis,
and
urine
chemistry
tests
from
10
animals/
sex
in
Groups
1
through
5.
Blood
was
collected
for
cholesterol
and
triglyceride
determinations
from
all
animals
prior
to
the
terminal
sacrifice
during
Week
105
(
Week
103
for
females
in
Group
3)
and
the
recovery
sacrifice
(
Group
6)
during
Week
106.
Blood
films
were
also
prepared
for
animals
at
the
terminal
and
recovery
sacrifices.
Five
animals/
sex
in
Groups
1
through
5
were
sacrificed
during
Week
4;
livers
were
collected
and
weighed.
[
The
liver
samples
were
collected
for
PFOS
analysis,
mitochondrial
activity,
hepatocellular
proliferation
rate
measurements
by
proliferation
cell
nuclear
antigen
(
PCNA),
and
determination
of
palmitoyl­
CoA
oxidase
activity.
In
addition,
liver
samples
were
collected
for
PFOS
analysis
(
Weeks
14,
53,
103,
105,
and
106),
hepatocellular
proliferation
rate
measurement
[
by
PCNA
at
Week
14
and
by
bromodeoxyuridine
(
BrdU)
immunohistochemistry
at
Week
53]
and
palmitoyl­
CoA
oxidase
determination
(
Week
4
and
14).]

RESULTS
Survival
rates:
There
was
a
significant
increased
trend
in
survival
that
occurred
in
the
males
that
was
due
to
significant
increases
in
survival
in
mid­
high
(
5.0
ppm)
and
high­
dose
(
20.0
ppm)
groups
as
compared
to
that
of
the
control
group.
None
of
the
other
treated
groups
in
the
males
revealed
any
significant
differences
in
survival.
No
significant
trend
was
noted
in
survival
in
females.
There
was
a
significant
decrease
in
survival
in
the
mid­
dose
(
2.0
ppm)
group
and
not
in
the
mid­
high
(
5.0
ppm)
and
high­
dose
(
20.0
ppm)
groups
as
compared
to
that
of
the
control.

Neoplastic
effects:

The
results
of
the
study
show
that
PFOS
is
carcinogenic,
inducing
tumors
of
the
liver,
and
of
the
thyroid
and
mammary
glands.
ENV/
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2002)
17/
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329
Tumor
incidence
(%)
Tumors
Male
0
0.5
ppm
2
ppm
5
ppm
20
ppm
20
ppm
recovery#
Liver
Hepatocellular
adenoma*
0
(
0/
60)
6.0
(
3/
50)
6.0
(
3/
50)
2.0
(
1/
50)
11.7**
(
7/
60)
Thyroid
Follicular
cell
adenoma
Follicular
cell
carcinoma
Combined
5.0
(
3/
60)
5.0
(
3/
60)
10.0
(
6/
60)
10.2
(
5/
49)
2.0
(
1/
49)
12.2
(
26/
49)
8.0
(
4/
50)
2.0
(
1/
50)
10.0
(
5/
50)
8.2
(
4/
49)
4.1
(
2/
49)
10.2
(
5/
49)
6.8
(
4/
59)
1.7
(
1/
59)
8.5
(
5/
59)
23.1**
(
9/
39)
2.6
(
1/
39)
25.6
(
10/
39)

Female
Liver
Hepatocellular
adenoma*
Hepatocellular
carcinoma
Combined*
0
(
0/
60)
0
(
0/
60)
0
(
0/
60)
2.0
(
1/
50)
0
(
0/
50)
2.0
(
1/
50)
2.0
(
1/
49)
0
(
0/
49)
2.0
(
1/
49)
2.0
(
1/
50)
0
(
1/
50)
2.0
(
1/
50)
8.3**
(
5/
60)
1.7
(
1/
60)
10.0**
(
6/
60)
Thyroid
Follicular
cell
adenoma
Follicular
cell
carcinoma
Combined
0
(
0/
60)
0
(
0/
60)
0
(
0/
60)
0
(
0/
50)
0
(
0/
50)
0
(
0/
50)
0
(
0/
49)
0
(
0/
49)
0
(
0/
49)
4.0
(
2/
50)
2.0
(
1/
50)
6.0**
(
3/
50)
1.7
(
1/
60)
0
(
0/
60)
1.7
(
1/
60)
Mammary
Fibroadenoma/
adenoma
Carcinoma
Combined
38.3
(
23/
60)
18.3
(
11/
60)
48.3
(
29/
60)
60.0**
(
30/
50)
24.0
(
12/
50)
72.0**
(
36/
50)
45.8
(
22/
48)
31.2
(
15/
48)
64.6**
(
31/
48)
52.04
(
26/
50)
22.0
(
11/
50)
58,0
(
29/
50)
25
(
15/
60)
23.3
(
14/
60)
40.0
(
24/
60)

*
Significant
positive
trend
(
P
<
0.03).
**
Significantly
increased
over
the
control
(
P
<
0.05).
#
Recovery
group;
after
52
weeks
of
treatment.

Remarks:
At
terminal
of
the
studies,
mean
body
weights
of
the
surviving
males
and
females
were
not
significantly
different
from
the
controls.
Food
consumption
for
males
and
females
was
similar
in
all
treated
groups
compared
to
animals
given
the
control
material
except
for
the
high­
dose
females
which
had
statistically
significantly
lower
food
consumption
during
weeks
2
through
44.
Except
for
the
mid­
dose
(
2.0
ppm)
female
group,
there
were
no
significant
decreases
in
survival
in
any
other
male
and
female
treatment
groups
as
compared
to
that
of
the
control.
Therefore,
it
appears
that
higher
dose
levels
could
have
been
tolerated
by
the
high­
dose
groups.

Nonneoplastic
effects:
NOAEL
(
dose
and
effect):
0.5
ppm
(
Male)
2
ppm
(
Female)
LOAEL
(
dose
and
effect):
2
ppm
(
Male)
5
ppm
(
Female)

Hepatotoxicity,
characterized
by
significant
increases
in
centrilobular
hypertrophy,
centrilobular
eosinophilic
hepatocytic
granules,
centrilobular
hepatocytic
pigment,
or
centrilobular
hepatocytic
ENV/
JM/
RD(
2002)
17/
FINAL
330
vacuolation
was
noted
in
male
and/
or
female
rats
given
5
or
20
ppm.
A
significant
increase
in
hepatocellular
centrilobular
hypertrophy
was
also
observed
in
mid­
dose
(
2
ppm)
male
rats.
Significant
increases
in
the
incidence
of
cystic
hepatocellular
degeneration
was
found
in
all
the
male
treated
groups
(
0.5,
2,
5,
or
20
ppm);
however,
this
liver
lesion
is
believed
to
be
due
to
old
age
of
the
animals
and
is
not
considered
to
be
treatment­
related.
Therefore,
based
on
the
pathological
findings
in
the
liver,
the
noobserved
adverse­
effect
level
(
NOAEL)
for
PFOS
is
considered
to
be
0.5
ppm
in
male
rats
and
2
ppm
in
female
rats;
the
low
observed­
adverse­
effect
level
(
LOAEL)
is
considered
to
be
2
ppm
in
male
rats
and
5
ppm
in
female
rats.

Results
of
Statistical
Analyses
of
Nonneoplastic
Lesions
in
Male
Rats.

Group
1
Control
2
Low
3
Mid
4
Mid­
High
5
High
6
High
Recovery
Groups
5
vs.
6
Liver
 
Vacuolation,
Hepatocellular
Midzonal/
Centrilobular
0/
65
0/
55
6/
55
10/
55
19/
55
9/
40
p­
values
.0000+**
NA
.1690
.0024+**
.0001+**
.4152+
0.0060+**

Liver
 
Hypertrophy,
Hepatocellular
Centrilobular
0/
65
2/
55
4/
55
22/
55
42/
565
3/
40
p­
values
.0000+**
.2080+
.0415+*
.0000+**
.0000+**
.0527+
.0000­**

Liver
 
Granular
Cytoplasm,
Eosinophilic,
Centrilobular
0/
65
0/
55
0/
55
0/
55
14/
65
0/
40
p­
values
.0000+**
NA
NA
BA
.0139+*
NA
.0007­**

Liver
 
Pigment
Hepatocellular,
Centrilobular
0/
65
0/
55
0/
55
0/
55
6/
65
0/
40
­
values
.006+**
NA
NA
NA
.0139+*
NA
.0513
Liver
 
Necrosis,
Individual
Hepatocyte
5/
65
4/
55
6/
55
5/
55
14/
65
4/
40
p­
values
.0106+*
NA
NA
NA
.0224+*
.0699+
.1024
Liver
 
Degeneration,
Cystic
5/
65
15/
55
19/
55
17/
55
22/
65
15/
40
NA
p­
values
.0007+**
.0041+**
.0003+**
.0011+**
.002+**
.0002+**
.4306+
ENV/
JM/
RD(
2002)
17/
FINAL
331
Results
of
Statistical
Analyses
of
Nonneoplastic
Lesions
in
the
Female
Rats.

1
2
3
4
Group
Control
Low
Mid
Mid­
High
5
High
6
High
Recovery
Groups
5
vs.
6
Liver
­
Infiltrate,
Lymphohistiocytic
42/
65
42/
55
38/
55
41/
55
p­
values
.0080
+**
NA
.3738+
NA
56/
65
.0038
+**
32/
40
.0709+
.2852
Liver
 
Hypertrophy,
Hepatocellular,
Centrilobular
2/
65
1/
55
4/
55
16/
55
p­
values
.0000
+**
NA
.2641+
.0001
+**
52/
65
.0000
+**
2/
40
NA
.0000
­**

Liver
­
Granular
Cytoplasm,
Eosinophilic,
Centrilobular
0/
65
0/
55
0/
55
7/
55
p­
values
.0000
+**
NA
NA
.0034
+**
36/
65
.0000
+**
1/
40
.3810+
.0000
­**

Liver
­
Pigment,
Hepatocellular,
Centrilobular
0/
65
0/
55
0/
55
1/
55
p­
values
.0000
+**
NA
NA
NA
36/
65
.0000
+**
0/
40
NA
.0000
­**

Liver
­
Necrosis,
Individual
Hepatocyte
7/
65
6/
55
6/
55
6/
55
p­
values
.0359
+*
NA
NA
NA
15/
65
.0500
+*
3/
40
.4254­
.0329
­*

Liver
 
Degeneration,
Cystic
0/
65
1/
55
1/
55
2/
55
p­
values
.0187
+*
NA
NA
.2080+
4/
65
.0596+
1/
40
.3810+
.3660
Liver
­
Hypertrophy,
Hepatocellular,
Periportal
12/
65
10/
55
9/
55
4/
55
p­
values
.0026
­**
.5796­
.4778­
.0614­
3/
65
.0127
­*
7/
40
NA
.0344
+*

Liver
­
Infiltrate,
Macrophage,
Pigmented
2/
65
3/
55
5/
55
6/
55
p­
values
.0000
+**
NA
.1567+
.0889
23/
65
.0000
+**
7/
40
.0147
+*
.0383
­*
ENV/
JM/
RD(
2002)
17/
FINAL
332
List
of
statistically
different
non­
neoplastic
effects
(
increased
compared
with
controls,
p
<
0.05):

Males
(
0.5ppm):
Liver
­
Degeneration,
Cystic
Males
(
2
ppm):
Liver
­
Degeneration,
Cystic
Liver
­
Hypertrophy,
Hepatocellular
Centrilobular
Males
(
5
ppm):
Liver
­
Degeneration,
Cystic
Liver
­
Hypertrophy,
Hepatocellular
Centrilobular
Liver
­
Vacuolation,
Hepatocellular
Midzonal
/
Centrilobular
Males
(
20
ppm):
Liver
­
Degeneration,
Cystic
Liver
­
Hypertrophy,
Hepatocellular
Centrilobular
Liver
­
Vacuolation,
Hepatocellular
Midzonal
/
Centrilobular
Liver
­
Granular
Cytoplasm,
Eosinophilic,
Centrilobular
Liver
­
Pigment
Hepatocellular,
Centrilobular
Liver
­
Necrosis,
Individual
Hepatocyte
Females
(
0.5
ppm):
None
Females
(
2
ppm):
None
Females
(
5
ppm):
Liver
­
Hypertrophy,
Hepatocellular
Centrilobular
Liver
­
Granular
Cytoplasm,
Eosinophilic,
Centrilobular
Liver
 
Infiltrate,
Macrophage,
Pigmented
Females
(
20
ppm):
Liver
­
Hypertrophy,
Hepatocellular
Centrilobular
Liver
­
Granular
Cytoplasm,
Eosinophilic,
Centrilobular
Liver
­
Infiltrate,
Macrophage,
Pigmented
Liver
­
Pigment
Hepatocellular,
Centrilobular
Liver
­
Necrosis,
Individual
Hepatocyte
Liver
­
Infiltrate,
Lymphohistiocytic
Liver
­
Hypertrophy,
Hepatocellular,
Periportal
Remarks:

The
cyctic
degeneration
of
the
liver
is
believed
to
be
due
to
old
age
of
the
animals
and
is
not
considered
to
be
treatment­
related.
However,
A
significant
increase
in
hepatocellular
centrilobular
hypertrophy
was
also
observed
in
mid­
dose
(
2
ppm)
male
rats.
Therefore,
the
NOAEL
for
the
male
rat
is
considered
to
be
0.5
ppm.
The
author
concluded
that
the
NOAEL
for
both
male
and
female
rats
are
2
ppm
(
which
we
do
not
agree).

There
was
no
effect
on
hepatic
palmitoyl­
CoA
oxidase
activity.
There
were
also
no
statistically
significant
increases
in
cell
proliferation
as
measured
by
proliferative
cell
nuclear
antigen
(
PCNA)
at
weeks
4
and
14,
or
by
bromodeoxyuridine
(
BrdU)
at
week
53.
ENV/
JM/
RD(
2002)
17/
FINAL
333
Serum
and
Liver
level
of
PFOS
Under
the
conditions
of
the
studies,
PFOS
was
observed
in
the
serum
and
liver
of
rats
dosed
with
perfluorooctane
sulfonic
acid
potassium
salt
(
PFOS
T­
6295).
Trace
levels
of
PFOS
were
often
detected
in
the
serum
and
liver
of
the
control
animals.

Summary
of
PFOS
Concentration­
Serum
(
ug/
mL)

a
Not
corrected
for
purity
of
the
standard
material.
b
LOQ­
Limit
of
Quanfitation
=
0.00910
pg/
mL
c
LOQ­
Limit
of
Quanfitation
=
0.0457
pg/
mL
d
C92987F
sample
spilled
during
extraction,
no
sample
remaining
for
analysis.
It
is
not
possible
to
verify
true
recovery
of
endogenous
analyte
from
tissues
without
radio­
labeled
reference
material.
The
only
measurement
of
accuracy
available
at
this
time,
matrix
spike
studies,
indicated
(
that
the
sera
data
are
accurate
to
±
30%;
liver
data
are
accurate
to
±
50%.
Group
6
Timepoint
Sex
0
ppm
0.5
ppm
2
ppm
5
ppm
20
ppm
High
Recovery
Average
±
SD
Average
±
SD
Average
±
SD
Average
±
SD
Average
±
SD
Average
±
SD
Male
<
LOQb
(
n=
5)
0.907
±
0.0619
(
n=
5)
4.33
±
1.16
(
n=
5)
7.57
±
2.17
(
n=
5)
41.8
±
7.92
(
n=
5)
Week
0
Female
0.0259
±
0.00663
(
n
=
5)
1.61
±
0.207
(
n
=
5)
6.62
±
0.499
(
n
=
5)
12.6
±
1.73
(
n
=
5)
54.0
±
7.34
(
n
=
5)

Male
<
LOQc
(
n
=
5)
4.04
±
0.801
(
n
=
5)
17.1
±
1.22
(
n
=
5)
43.9
±
4.90
(
n
=
5)
148
±
13.8
(
n
=
5)
Week
14a
Female
2.67
±
4.58
(
n
=
5)
6.96
±
0.993
(
n
=
4d)
27.3
±
2.34
(
n
=
5)
64.4
±
5.48
(
n
=
5)
223
±
22.4
(
n
=
5)

Male
0.0249
±
0.0182
(
n
=
5)
146
±
33.5
(
n
=
4)
Week
53
Female
0.395
±
0.777
(
n
=
5)
220
±
44.0
(
n
=
5)

Male
Day
719
Female
20.2
±
13.3
(
n
=
9)

Male
0.0118
±
0.0104
(
n
=
11)
1.31
±
1.30
(
n
=
10)
7.60
±
8.60
(
n
=
17)
22.5
±
23.5
(
n
=
25)
69.3
±
57.9
(
n
=
22)
Week
105
Female
0.0836
±
0.134
(
n
=
24)
4.35
±
2.78
(
n
=
15)
75.0
±
45.7
(
n
=
15)
233
±
124
(
n
=
25)

Male
2.42
±
5.09
(
n
=
10)
Week
106
Female
9.51
±
8.70
(
n
=
17)
ENV/
JM/
RD(
2002)
17/
FINAL
334
Summary
of
PFOS
Concentration­
Liver
(
ug/
g)

Group
1
Group
2
Group
3
Group
4
Group
5
Group
6
Timepoint
Sex
Control
Low
Mid
Mid­
High
High
High
Recovery
Average
±
SD
Average
±
SD
Average
±
SD
Average
±
SD
Average
±
SD
Average
±
SD
Male
0.104
±
0.0673
(
n
=
5)
11.0
±
2.31
(
n
=
5)
31.3
±
5.84
(
n
=
5)
47.6
±
12.5
(
n
=
5)
282
±
45.3
(
n
=
5)
Week
0
Female
0.107
±
0.0486
(
n
=
5)
8.71
±
0.552
(
n
=
5)
25.0
±
6.11
(
n
=
5)
83.0
±
14.1
(
n
=
5)
373
±
44.1
(
n
=
5)

Male
0.459
±
0.0573
(
n
=
5)
23.8
±
3.45
(
n
=
5)
74.0
±
6.16
(
n
=
5)
358
±
28.8
(
n
=
5)
568
±
107
(
n
=
5)
Week
10
Female
12.0
±
22.4
(
n
=
5)
19.2
±
3.77
(
n
=
5)
69.2
±
3.46
(
n
=
5)
370
±
22.3
(
n
=
5)
635
±
49.0
(
n
=
5)

Male
0.635
±
1.04
(
n
=
10)
435
±
96.9
(
n
=
9)
Week
53
Female
0.923
±
1.77
(
n
=
10)
560
±
180
(
n
=
10)

Male
Day
719
Female
55.1
±
31.5
(
n
=
9)

Male
0.114
±
0.148
(
n
=
11)
7.83
±
7.34
(
n
=
10)
26.4
±
20.4
(
n
=
17)
70.5
±
63.1
(
n
=
25)
189
±
141
(
n
=
22)
Week
105
Female
0.185
±
0.184
(
n
=
24)
12.9
±
6.81
(
n
=
15)
131
±
61.4
(
n
=
15)
381
±
176
(
n
=
25)

Male
3.12
±
5.97
(
n
=
10)
Week
106
Female
12.9
±
10.4
(
n
=
17)



It
is
not
possible
to
verify
true
recovery
of
endogenous
analyte
from
tissues
without
radio­
labeled
reference
material.
The
only
measurement
of
accuracy
available
at
this
time,
matrix
spike
studies,
indicated
that
the
sera
data
are
accurate
to
±
30%;
liver
data
are
accurate
to
±
50%.

CONCLUSIONS
The
study
results
are
summarized
as
follows:
1.
Treatment­
related
changes
were
found
more
commonly
in
males
than
in
females
of
each
of
the
treatment
groups,
which
were
supported
by
earlier
pharmacokinetic
studies
demonstrating
a
higher
retention
of
the
compound
by
males
than
females.
2.
The
test
material
was
considered
to
be
carcinogenic
in
the
rat,
inducing
tumors
of
the
liver
and
the
thyroid
gland
in
the
males
and
tumors
of
the
liver
and
of
the
thyroid
and
mammary
gland
in
females.
3.
Based
on
the
pathological
findings
in
the
liver,
the
no­
observed­
adverse­
effect
level
(
NOAEL)
for
PFOS
is
considered
to
be
0.5
ppm
in
male
rats
and
2
ppm
in
female
rats;
the
low
observedadverse
effect
level
(
LOAEL)
is
considered
to
be
2
ppm
in
male
rats
and
5
ppm
in
female
rats.
ENV/
JM/
RD(
2002)
17/
FINAL
335
4.
There
was
no
effect
on
hepatic
palmitoyl­
CoA
oxidase
activity.
There
were
also
no
statistically
significant
increases
in
cell
proliferation
as
measured
by
proliferative
cell
nuclear
antigen
(
PCNA)
or
by
bromodeoxyuridine
(
BrdU).
5.
PFOS
was
observed
in
the
serum
and
liver
of
rats
dosed
with
perfluorooctane
sulfonic
acid
potassium
salt
(
PFOS
T­
6295).
Trace
levels
of
PFOS
were
often
detected
in
the
serum
and
liver
of
the
control
animals.

REFERENCE
3M,
(
2002).
104­
Week
Dietary
Chronic
Toxicity
and
Carcinogenicity
Study
with
Perfluorooctane
Sulfonic
Acid
Potassium
Salt
(
PFOS;
T­
6295)
in
Rats.
Final
Report,
3M
T­
6295
(
Covance
study
no.:
6329­
183),
Volumes
I­
IX
,
4068
pages,
January
2,
2002.
3M,
St.
Paul,
Minnesota.

Seacar,
A.
M.,
Thomford,
P.
J.,
and
Butenhoff,
J.
L.
Terminal
observations
in
Sprague­
Dawley
rats
after
lifetime
dietary
exposure
to
potassium
perfluorooctanesulfonate.
Toxicol.
Sci./
Toxicologist,
66
(
1­
S):
185,
2002.
ENV/
JM/
RD(
2002)
17/
FINAL
336
EPIDEMIOLOGIC
DATA
Title:
Identification
of
Fluorochemicals
in
Sera
of
American
Red
Cross
Adult
Blood
Donors
TEST
SUBSTANCE
Identity:
PFOS
and
6
other
fluorochemicals
Remarks:
The
results
reported
are
preliminary.
The
final
report
is
expected
November
2001.

METHOD
Study
design:
Cross­
sectional
Manufacturing/
Processing/
Use:
N/
A
Hypothesis
tested:
To
determine
the
levels
of
PFOS
in
the
serum
of
American
Red
Cross
blood
banks
in
6
regions
of
the
U.
S.

Study
period:
2000
Setting:
N/
A
Total
population:
Serum
pooled
from
6
ARC
blood
banks
in
various
geographic
regions
in
the
US:
Los
Angeles,
CA;
Minneapolis/
St.
Paul,
MN;
Charlotte,
NC;
Boston,
MA;
Portland,
OR,
and
Hagerstown,
MD.

Subject
selection
criteria:
Unknown
Total
#
of
subjects
in
study:
652
donors,
age
20­
69
years
Comparison
population:
N/
A
Participation
rate:
N/
A
Subject
description:
No
information
was
provided
on
the
individuals
from
whom
the
sera
samples
were
taken.

Health
effects
studied:
PFOS
levels
in
blood
Data
collection
methods:
Blood
sera
samples
were
analyzed
using
high­
pressure
liquid
chromatography/
electrospray
tandem
mass
spectrometry
(
HPLC/
ESMSMS).

Details
on
data
collection:
No
information
was
provided
as
to
how
the
blood
was
drawn,
stored,
etc.

Exposure
period:
Unknown­­
PFOS
serum
levels
used
as
surrogate
for
exposure.

Description/
delineation
of
exposure
groups/
categories:
N/
A
Measured
or
estimated
exposure:
N/
A
ENV/
JM/
RD(
2002)
17/
FINAL
337
Exposure
levels:
N/
A
Statistical
methods:
Arithmetic
means,
ranges,
geometric
means
and
95%
confidence
intervals
were
calculated.
Central
tendency
and
distribution
of
the
data
by
age,
gender,
location
and
their
respective
interaction
terms
will
be
done
in
the
final
report.
A
reliability
assessment
is
also
being
analyzed.

Other
methodological
information:
N/
A
RESULTS
Describe
results:
The
mean
serum
PFOS
level
was
43.7
ppb.
The
range
was
4.27
to
1656
ppb.
Analyses
stratified
by
age,
gender,
and
geographic
location
will
be
forthcoming
in
the
final
report.

Study
strengths
and
weaknesses:
These
data
are
cross­
sectional
data
used
to
determine
PFOS
levels
in
the
general
population.
No
other
descriptive
information
about
the
subjects
is
available
in
this
preliminary
report.
The
sample
size
is
relatively
small.
Blood
donors
cannot
be
considered
representative
of
the
general
population
of
the
US.

Research
sponsors:
3M
Medical
Department,
Corporate
Occupational
Medicine
Consistency
of
results:
Mean
PFOS
levels
reported
in
this
study
are
similar
to
those
reported
earlier
in
pooled
blood
samples,
although
they
are
on
the
higher
end
of
the
range.

CONCLUSIONS
N/
A
REFERENCE
Olsen,
GW,
Burris,
JM,
Lundberg,
JK,
Hansen,
KJ,
Mandel,
JH,
Zobel,
LR.
Identification
of
fluorochemicals
in
sera
of
American
Red
Cross
adult
blood
donors.
Interim
report.
June
25,
2001.
ENV/
JM/
RD(
2002)
17/
FINAL
338
EPIDEMIOLOGIC
DATA
Title:
An
Epidemiologic
Analysis
of
Episodes
of
Care
of
3M
Decatur
Chemical
and
Film
Plant
Employees,
1993­
1998
TEST
SUBSTANCE
Identity:
POSF­
based
chemicals
used
at
the
Decatur
plant
Remarks:
Episodes
of
care
analyses
are
not
often
used
in
occupational
epidemiologic
studies.

METHOD
Study
design:
Episode
of
care
comparison
Manufacturing/
Processing/
Use:
The
3M
Decatur,
Alabama
plant
began
production
in
1961.
It
is
made
up
of
the
film
plant
and
the
chemical
plant.
The
3
major
product
groups
in
the
chemical
plant
are
protective
chemicals,
performance
chemicals,
and
fluoroelastomers.
Perfluorooctanesulfonyl
fluoride
(
POSF)
is
the
major
sulfonate
fluorochemical
manufactured
at
Decatur
and
is
used
as
the
precursor
to
the
production
of
a
variety
of
perfluorinated
amides,
alcohols,
acrylates,
and
other
fluorochemical
polymers.

Hypothesis
tested:
To
use
episodes
of
care
methodology
as
a
screen
for
morbidity
outcomes
associated
with
long­
term,
high
exposure
to
POSF­
based
production
at
the
3M
facility
in
Decatur,
Alabama.

Study
period:
Episodes
of
care
experience
of
652
chemical
employees
and
659
film
plant
employees
were
analyzed
for
workers
at
the
plant
who
were
employed
for
at
least
1
year
between
January
1,
1993
and
December
31,
1998.

Setting:
3M
plant
in
Decatur,
Alabama.

Total
population:
1311
workers
were
eligible
for
the
cohort
(
at
least
1
year
of
employment
at
the
plant).
The
total
worker
population
was
not
reported.

Subject
selection
criteria:
All
workers
employed
at
the
Decatur
plant
for
at
least
1
year
between
Jan.
1,
1993
and
Dec.
31,
1998.
Episodes
of
care
were
limited
to
their
Decatur
time
of
employment
for
employees
hired,
terminated,
or
died
during
the
study
period.
However,
records
of
employees
on
Medicare,
long­
term
disability
or
who
chose
HMO
coverage
were
not
in
the
database
and
would
not
be
included
in
the
episodes
of
care
for
that
employee.

Comparison
population:
Chemical
and
film
plant
employees
were
analyzed
separately
and
then
compared
to
each
other.
Employee
comparison
groups
were
defined
according
to
their
potential
workplace
exposure
to
POSF
fluorochemical
production.
Group
A:
all
chemical
plant
employees
and
all
film
plant
employees
eligible
for
the
cohort.
Group
B:
all
chemical
plant
employees
who
worked
solely
in
the
chemical
plant
and
all
film
plant
employees
who
worked
exclusively
in
the
film
plant.
Group
C:
all
chemical
plant
employees
with
high
fluorochemical
exposures
compared
to
their
job
counterparts
in
the
film
plant.
Group
D:
all
plant
workers
with
high
fluorochemical
exposure
for
at
least
10
years
prior
to
the
study
onset
compared
to
their
job
counterparts
in
the
film
plant.

Participation
rate:
97%
of
Decatur
employees
were
eligible
for
participation
in
the
study.
ENV/
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2002)
17/
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339
Subject
description:
82%
of
the
employees
in
the
cohort
were
male
(
530
in
the
chemical
plant
and
558
in
the
film
plant).
The
mean
age
was
45.1
in
the
chemical
plant
and
48.6
in
the
film
plant.
Sixty
percent
of
the
chemical
plant
employees
had
worked
only
in
the
chemical
plant
and
a
similar
percentage
of
film
plant
workers
had
worked
exclusively
in
the
film
plant.
Seventy­
six
percent
of
the
chemical
plant
workers
had
high
exposure
jobs.

Health
effects
studied:
Morbidity.
Based
on
animal
data
and
epidemiologic
studies
on
PFOA
and
PFOS,
certain
episodes
of
care
were
considered
a
priori.
They
included:
liver
and
bladder
cancer,
endocrine
disorders
involving
the
thyroid
gland
and
lipid
metabolism,
gastrointestinal
disorders
of
the
liver
and
biliary
tract,
and
reproductive
disorders.

Data
collection
methods:
The
Clinical
Care
Groups
episode
of
care
software
developed
by
Ingenix,
Inc.
was
used
to
provide
a
comprehensive
grouping
of
all
visits
(
inpatient
and
outpatient),
procedures,
ancillary
services,
and
prescription
drugs
used
in
the
diagnosis,
treatment
and
management
of
more
than
400
diseases
or
conditions.
The
software
code
constructs
an
episode
of
care
around
the
index­
eligible
record
by
searching
backward
and
forward
in
time
for
the
health
claims
records
that
are
related
to
the
disease
or
condition
on
the
index
record.
The
index
record
consists
of
either
procedure
codes
indicative
of
a
face­
toface
encounter
or
a
pharmacy
record
for
a
delineating
drug.

Exposure
period:
The
episodes
of
care
that
were
included
in
the
study
were
those
experienced
between
Jan.
1,
1993
to
Dec.
31,
1998.

Description/
delineation
of
exposure
groups/
categories:
Workers
were
placed
into
groups
according
to
potential
workplace
exposures:
workers
who
were
employed
solely
in
the
chemical
or
film
plants,
those
who
had
high
exposure
jobs,
and
those
who
worked
at
least
10
years
in
jobs
with
high
potential
for
fluorochemical
exposure.

Measured
or
estimated
exposure:
estimated
based
on
job
history
information.

Exposure
levels:
Not
measured.
Employees
were
placed
into
exposure
categories
based
on
job
description.

Statistical
methods:
A
risk
ratio
episode
of
care
(
RREpC)
provided
the
estimate
of
risk
between
the
observed
to
expected
episodes
of
care
for
chemical
plant
employees
compared
to
the
observed
to
expected
episodes
of
care
among
film
plant
employees.
The
expected
number
of
episodes
of
care
for
both
the
film
and
chemical
plant
employees
was
calculated
from
health
claims
data
of
the
3M
manufacturing
population
in
the
U.
S.
Because
the
chemical
and
film
plant
cohorts
had
slightly
different
age
and
gender
structures,
an
adjusted
ratio
was
calculated
and
compared
to
the
unadjusted
risk
ratio.
In
most
cases,
the
risk
ratios
were
comparable.
Therefore,
95%
confidence
intervals
were
only
calculated
for
the
unadjusted
risk
ratios.

Other
methodological
information:
It
should
be
noted
that
from
an
epidemiologic
perspective,
an
episode
of
care
could
represent
any
and
all
incident
cases,
prevalent
cases,
and/
or
misclassified
cases
(
both
false
positive
and
false
negative).
In
addition,
types
and
counts
of
episodes
of
care
may
differ
by
the
software
used,
and
it
is
possible
that
2
different
diagnoses
may
be
assigned
to
the
same
episode.
Certain
services,
such
as
lab
procedures
and
prescriptions
may
not
be
reported
for
the
episode.
Also,
the
endpoint
of
an
episode
may
vary
among
software
programs.
The
clinical
flexibility
of
the
algorithm
may
differ
depending
on
the
software
program.

RESULTS
Describe
results:
The
only
increased
risk
of
episodes
for
the
conditions
of
a
priori
interest
were
for
ENV/
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2002)
17/
FINAL
340
neoplasms
of
the
male
reproductive
system
and
for
the
overall
category
of
cancers
and
benign
growths
(
which
included
cancer
of
the
male
reproductive
system).
There
was
an
increased
risk
of
episodes
for
the
overall
cancer
category
for
all
4
comparison
groups.
The
risk
ratio
was
greatest
in
the
group
of
employees
with
the
highest
and
longest
exposures
to
fluorochemicals
(
RREpC
=
1.6,
95%
CI
=
1.2
 
2.1).
Increased
risk
of
episodes
in
long­
time,
high­
exposure
employees
also
was
reported
for
male
reproductive
cancers
(
RREpC
=
9.7,
95%
CI
=
1.1
­
458).
It
should
be
noted
that
the
confidence
interval
is
very
wide
for
male
reproductive
cancers
and
the
sub­
category
of
prostate
cancer.
Five
episodes
of
care
were
observed
for
reproductive
cancers
in
chemical
plant
employees
(
1.8
expected),
of
which
4
were
prostate
cancers.
One
episode
of
prostate
cancer
was
observed
in
film
plant
employees
(
3.4
expected).
This
finding
is
important
because
an
excess
in
prostate
cancer
mortality
was
observed
in
the
Cottage
Grove
plant
mortality
study.
However,
the
update
of
the
study
did
not
confirm
this
finding.

There
was
an
increased
risk
of
episodes
for
neoplasms
of
the
gastrointestinal
tract
in
the
high
exposure
group
(
RREpC
=
1.8,
95%
CI
=
1.2­
3.0)
and
the
long­
term
employment,
high
exposure
group
(
RREpC
=
2.9,
95%
CI
=
1.7
 
5.2).
Most
of
the
episodes
were
attributable
to
benign
colonic
polyps.
Similar
numbers
of
episodes
were
reported
in
film
and
chemical
plant
employees.

In
the
entire
cohort,
only
1
episode
of
care
was
reported
for
liver
cancer
(
0.6
expected)
and
1
for
bladder
cancer
(
1.5
expected).
Both
occurred
in
film
plant
employees.
Only
2
cases
of
cirrhosis
of
the
liver
were
observed
(
0.9
expected),
both
in
the
chemical
plant.
There
was
a
greater
risk
of
lower
urinary
tract
infections
in
chemical
plant
employees,
but
they
were
mostly
due
to
recurring
episodes
of
care
by
the
same
employees.
It
is
difficult
to
draw
any
conclusions
about
these
observations,
given
the
small
number
of
episodes
reported.

Chemical
plant
employees
in
the
high
exposure,
long­
term
employment
group
were
2
½
times
more
likely
to
seek
care
for
disorders
of
the
biliary
tract
than
their
counterparts
in
the
film
plant
(
RREpC
=
2.6,
95%
CI
=
1.2
­
5.5).
Eighteen
episodes
of
care
were
observed
in
chemical
plant
employees
and
14
in
film
plant
workers.
The
sub­
categories
that
influenced
this
observation
were
episodes
of
cholelithiasis
with
acute
cholecystitis
and
cholelithiasis
with
chronic
or
unspecified
cholecystitis.
Most
of
the
observed
cases
occurred
in
chemical
plant
employees.

Risk
ratios
of
episodes
of
care
for
endocrine
disorders,
which
included
sub­
categories
of
thyroid
disease,
diabetes,
hyperlipidemia,
and
other
endocrine
or
nutritional
disorders,
were
not
elevated
in
the
comparison
groups.
Conditions
which
were
not
identified
a
priori
but
which
excluded
the
null
hypothesis
in
the
95%
confidence
interval
for
the
high
exposure,
long­
term
employment
group
included:
disorders
of
the
pancreas,
cystitis,
and
lower
urinary
tract
infections.

Study
strengths
and
weaknesses:
See
"
other
methodological
information"
section
for
limitations
of
episodes
of
care
software.
The
results
of
this
study
should
only
be
used
for
hypothesis
generation.
Although
the
episode
of
care
design
allowed
for
a
direct
comparison
of
workers
with
similar
demographics
but
different
exposures,
there
are
many
limitations
to
this
design.
Episodes
of
care
are
reported,
not
disease
incidence;
therefore,
this
parameter
cannot
be
interpreted
in
any
other
manner.
The
data
are
difficult
to
interpret
because
a
large
RREpC
may
not
necessarily
indicate
high
risk
of
incidence
of
disease.
In
addition,
many
of
the
risk
ratios
for
episodes
of
care
had
very
wide
confidence
intervals.
The
analysis
was
limited
to
6
years.
Also,
the
utilization
of
health
care
services
may
reflect
local
medical
practice
patterns.
Individuals
may
be
counted
more
than
once
in
the
database
because
they
can
be
categorized
under
larger
or
smaller
disease
classifications.
Episodes
of
care
may
include
the
same
individual
several
times.
Not
all
employees
were
included
in
the
database,
such
as
those
on
long­
term
disability.

Research
sponsors:
3M
Company
ENV/
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RD(
2002)
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341
Consistency
of
results:
No
other
morbidity
studies
have
been
conducted
on
fluorochemicals.

CONCLUSIONS
This
study
should
only
be
used
for
hypothesis
generation
regarding
workers
employed
at
the
Decatur
plant
who
are
employed
in
jobs
with
high
exposure
to
POSF­
based
fluorochemicals.

REFERENCE
Olsen,
GW,
Burlew,
MM,
Hocking,
BB,
Skratt,
JC,
Burris,
JM,
Mandel,
JH.
An
epidemiologic
analysis
of
episodes
of
care
of
3M
Decatur
chemical
and
film
plant
employees,
1993­
1998.
Final
Report.
May
18,
2001.
ENV/
JM/
RD(
2002)
17/
FINAL
342
EPIDEMIOLOGIC
DATA
Title:
Identification
of
Fluorochemicals
in
Sera
of
Children
in
the
United
States
TEST
SUBSTANCE
Identity:
PFOS
and
6
other
fluorochemicals
Remarks:
The
results
reported
are
preliminary.
The
final
report
is
expected
November
2001.

METHOD
Study
design:
Cross­
sectional.

Manufacturing/
Processing/
Use:
N/
A
Hypothesis
tested:
To
determine
the
serum
concentrations
of
selected
fluorochemicals
in
a
sample
of
children
to
provide
a
more
specific
understanding
of
the
distribution
of
these
compounds
in
children.

Study
period:
Child
sera
samples
were
collected
from
January
1994
to
March
1995.
The
sera
samples
were
analyzed
in
Spring
1999.

Setting:
N/
A
Total
population:
Not
reported
Subject
selection
criteria:
The
sera
samples
were
provided
to
3M
by
the
University
of
Minnesota
Department
of
Pediatrics.
They
were
obtained
from
a
large
clinical
trial
on
Group
A
streptococcal
infections
in
children.
The
children
were
residents
of
23
states
in
the
US.
These
children
presented
with
signs
and
symptoms
of
acute­
onset
pharyngitis.
All
of
the
children
had
positive
throat
cultures
at
the
initial
visit.

Total
#
of
subjects
in
study:
n
=
599
children,
age
2­
12
years
Comparison
population:
N/
A
Participation
rate:
N/
A
Subject
description:
No
information
was
provided
on
the
children
from
whom
the
sera
samples
were
taken.

Health
effects
studied:
PFOS
serum
levels
in
blood,
as
well
as
6
other
fluorochemicals.

Data
collection
methods:
Blood
sera
samples
were
collected
using
high­
pressure
liquid
chromatography/
electrospray
tandem
mass
spectrometry
(
HPLC/
ESMSMS).
The
samples
were
collected
from
equal
numbers
of
male
and
female
children
residing
in
23
states.

Details
on
data
collection:
No
information
was
provided
as
to
how
the
blood
was
drawn,
stored,
etc.

Exposure
period:
N/
A
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Description/
delineation
of
exposure
groups/
categories:
Blood
sera
samples
were
collected
from
children
2
 
12
years
old.

Measured
or
estimated
exposure:
N/
A
Exposure
levels:
N/
A
Statistical
methods:
Arithmetic
means,
ranges,
geometric
means
and
95%
confidence
intervals
were
calculated.
Central
tendency
and
distribution
of
the
data
by
age,
gender,
location
and
their
respective
interaction
terms
will
be
done
in
the
final
report.
A
reliability
assessment
is
also
being
analyzed.

Other
methodological
information:
N/
A
RESULTS
Describe
results:
The
mean
PFOS
serum
level
was
43.5
ppb.
The
range
was
6.7
­
515
ppb.
Analyses
stratified
by
age,
gender,
and
geographic
location
will
be
forthcoming
in
the
final
report.

Study
strengths
and
weaknesses:
These
data
are
cross­
sectional
data
used
to
determine
PFOS
levels
in
U.
S.
children.
No
other
descriptive
information
about
the
subjects
is
available
in
this
preliminary
report.
The
sample
size
is
relatively
small.

Research
sponsors:
3M
Medical
Department,
Corporate
Occupational
Medicine
Consistency
of
results:
To
date,
no
other
data
have
been
collected
on
PFOS
serum
levels
in
children.

CONCLUSIONS
N/
A
REFERENCE
Olsen,
GW,
Burris,
JM,
Lundberg,
JK,
Hansen,
KJ,
Mandel,
JH,
Zobel,
LR.
Identification
of
fluorochemicals
in
sera
of
children
in
the
United
States.
Interim
Report.
June
25,
2001.
ENV/
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2002)
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EPIDEMIOLOGIC
DATA
Title:
Mortality
Study
of
Workers
Employed
at
the
3M
Decatur
Facility
TEST
SUBSTANCE
Identity:
Several
perfluorooctanesulfonyl
fluoride­
based
fluorochemicals,
including
PFOS
Remarks:
This
study
is
an
update
of
the
study
published
by
Mandel
and
Johnson,
1995.

METHOD
Study
design:
Retrospective
cohort
mortality
study.

Manufacturing/
Processing/
Use:
The
3M
Decatur,
Alabama
plant
began
production
in
1961.
It
is
made
up
of
the
film
plant
and
the
chemical
plant.
The
3
major
product
groups
in
the
chemical
plant
are
protective
chemicals,
performance
chemicals,
and
fluoroelastomers.
Perfluorooctanesulfonyl
fluoride
(
POSF)
is
the
major
sulfonate
fluorochemical
manufactured
at
Decatur
and
is
used
as
the
precursor
to
the
production
of
a
variety
of
perfluorinated
amides,
alcohols,
acrylates,
and
other
fluorochemical
polymers.

Hypothesis
tested:
To
determine
whether
occupational
exposure
to
fluorochemicals
is
related
to
mortality
of
employees
of
the
3M
facility
in
Decatur,
Alabama
Study
period:
The
study
population
worked
at
the
plant
for
at
least
1
year
since
it
began
production
in
1961.
The
cohort
was
followed
through
Dec.
31,
1997.
Currently
employed
workers
were
assigned
Dec
31,
1997
as
their
last
date
of
employment.

Setting:
3M
plant
in
Decatur,
Alabama.

Total
population:
3512
workers
were
identified.
Of
these
workers,
2083
worked
at
the
plant
for
at
least
one
year.
Eighty­
four
percent
of
the
cohort
was
male.

Subject
selection
criteria:
All
workers
employed
at
the
Decatur
plant
for
at
least
1
year.
The
cohort
was
followed
through
Dec.
31,
1997.
Currently
employed
workers
were
assigned
Dec
31,
1997
as
their
last
date
of
employment.

Comparison
population:
In
SMR
analyses,
Minnesota
population
death
rates
for
whites
were
used.
Mortality
reference
rates
from
7
regional
counties
were
also
used
to
rule
out
large
variations
based
on
regional
mortality
reporting
differences.

Participation
rate:
Death
certificates
were
obtained
for
96%
(
n
=
139)
of
the
cohort
who
were
deceased.

Subject
description:
84%
of
the
employees
in
the
cohort
were
male.
The
mean
age
at
follow­
up
was
51.1
years,
and
the
mean
number
of
years
worked
at
the
plant
was
14.9.
The
number
of
person­
years
at
followup
was
50970.
There
were
145
deaths
identified
in
the
cohort.
The
high
exposure
group
was
slightly
younger
than
the
other
2
groups
but
worked
3
years
longer
at
the
plant
than
the
other
2
exposure
groups.

Health
effects
studied:
Mortality
Data
collection
methods:
A
review
of
employee
work
history
records
of
any
employee
with
at
least
1
year
employment
were
abstracted
to
record
the
workers'
name,
SSN,
3M
identification
number,
date
of
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birth,
and
dates
of
work
history.
This
cohort
was
linked
to
records
from
the
original
cohort
to
update
the
employment
information
and
verify
other
data.
The
National
Death
Index
was
searched
for
all
of
the
workers.
Discrepancies
with
the
original
cohort
were
resolved
and
deaths
before
1979
were
verified
in
the
Social
Security
Death
Index.
A
licensed
nosologist
coded
the
death
certificates
to
ICD
8.

Exposure
period:
The
potential
exposure
period
was
from
the
plant
opening
in
1961
to
Dec.
31,
1997.

Description/
delineation
of
exposure
groups/
categories:
Workers
were
placed
into
3
exposure
groups
based
on
job
history
information:
high
exposure,
low
exposure,
and
no
exposure.
There
were
145
deaths
in
the
cohort:
65
in
the
high
exposure
group,
27
in
the
low
exposure
group
and
53
in
the
non­
exposed
group.

Measured
or
estimated
exposure:
estimated
based
on
job
history
information.

Exposure
levels:
Not
measured.
Employees
were
placed
into
3
exposure
categories
based
on
job
description:
low
exposure
(
n
=
289),
high
exposure
(
n
=
782),
non­
exposed
(
n
=
812).

Statistical
methods:
Standardized
Mortality
Ratios
(
SMRs)
and
95%
confidence
intervals
were
derived
using
the
PC
Life
Table
Analysis
System
software
developed
by
NIOSH.
This
program
computes
age,
gender,
and
race­
specific
SMRs
using
standard
life
table
methods.
The
expected
number
of
deaths
is
estimated
by
multiplying
the
age,
gender,
race,
and
calendar
period
tabulated
person­
years
of
follow
up
to
the
corresponding
cause­
specific
mortality
reference
rates.
Mortality
rates
for
white
Minnesotans
were
used
as
reference
data.

Other
methodological
information:

RESULTS
Describe
results:
145
deaths
were
identified
in
the
cohort:
65
of
these
deaths
were
in
the
high
exposure
group.
When
the
entire
cohort
was
analyzed,
SMRs
were
not
elevated
for
most
of
the
cancer
types
and
for
non­
malignant
causes.
SMRs
that
were
above
1
(
cancer
of
the
esophagus,
liver,
breast,
urinary
organs,
bladder,
and
skin)
were
also
elevated
when
the
cohort
was
limited
to
any
employee
ever
employed
in
a
high
exposure
job
(
except
breast
cancer).
Only
2
or
3
deaths
were
reported
for
each
of
these
cause­
specific
categories
and
were
not
statistically
significant,
except
for
bladder
cancer.

Workers
who
were
employed
in
high
exposure
jobs
were
greater
than
12
times
more
likely
to
die
of
bladder
cancer
than
the
general
population
of
Alabama
(
SMR
=
12.77,
95%
CI
=
2.63
­
37.35).
This
effect
remained
when
the
data
were
analyzed
using
county
death
rates.
Three
male
employees
in
the
cohort
died
of
bladder
cancer,
and
all
of
the
deaths
occurred
in
employees
who
had
worked
in
high
exposure
jobs
for
at
least
5
years
(
SMR
=
24.49,
3
observed
deaths,
0.12
expected).
All
of
them
had
worked
at
the
Decatur
plant
for
more
than
20
years.
In
the
previous
cohort
mortality
study,
1
bladder
cancer
death
was
reported.

Two
deaths
were
reported
for
liver
cancer.
One
was
in
the
low
exposure
group
and
one
in
the
high
exposure
group.
The
SMR
for
workers
who
were
employed
in
either
high
or
low
exposure
jobs
was
3.08
(
95%
CI
=
0.37
 
11.10).
Five
cases
of
cirrhosis
of
the
liver
were
reported
in
this
cohort,
2
in
the
high
exposure
group,
1
in
the
low
exposure
group,
and
2
in
the
non­
exposed.
The
observed
did
not
exceed
the
expected
mortality
experience
in
any
of
these
groups.

Study
strengths
and
weaknesses:
A
larger
cohort
was
followed
in
this
update
of
the
original
study,
thus
reporting
twice
as
many
deaths;
however,
the
cohort
is
fairly
young
and
the
number
of
deaths
is
still
small.
There
were
small
numbers
of
deaths
in
many
of
the
categories
for
males
and
especially
for
females
in
all
categories,
thus
limiting
the
power
of
the
study.
Death
certificates
were
located
for
96%
of
the
cohort,
but
ENV/
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the
6
not
obtained
could
greatly
impact
the
results
of
the
analyses
since
the
number
of
deaths
in
most
of
the
sub­
cohorts
was
very
small.
Biological
measurements
of
fluorochemicals
were
not
available;
therefore,
exposure
categories
based
on
job
descriptions
were
used
as
a
surrogate
for
exposure.
The
categories
are
more
specific
in
this
update
of
the
study;
however,
there
is
still
a
potential
for
misclassification
of
exposure.
Based
on
the
results
of
the
biomonitoring
conducted
on
employees
in
a
random
sample
of
Decatur
employees
in
1998,
the
geometric
mean
of
PFOS
in
film
plant
employees
was
0.136
ppm.
However,
in
this
study,
film
plant
employees
are
considered
non­
exposed.
In
addition
to
fluorochemicals,
workers
were
exposed
to
other
chemicals
in
the
workplace.

Research
sponsors:
University
of
Minnesota
Consistency
of
results:
In
the
first
study,
there
was
an
excess
of
bladder
cancer;
however,
this
was
based
on
only
1
death.
Three
additional
deaths
have
now
been
reported.

CONCLUSIONS
Workers
employed
at
the
Decatur
plant
who
are
employed
in
jobs
with
high
exposure
to
POSF­
based
fluorochemicals
are
at
increased
risk
of
death
from
bladder
cancer.

REFERENCE
Alexander,
B.
H.
April
26,
2001.
Mortality
study
of
workers
employed
at
the
3M
Decatur
facility.
Final
Report.
Division
of
Environmental
and
Occupational
Health,
School
of
Public
Health,
University
of
Minnesota.

OTHER
This
study
is
the
second
update
of
the
mortality
study.
ENV/
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2002)
17/
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EPIDEMIOLOGIC
DATA
Title:
Identification
of
Fluorochemicals
in
Human
Sera.
I.
American
Red
Cross
Adult
Blood
Donors
TEST
SUBSTANCE
Identity:
PFOS
and
6
other
fluorochemicals
Remarks:
This
is
a
final
report.
The
preliminary
report
was
dated
June
25,
2001.

METHOD
Study
design:
Cross­
sectional
Manufacturing/
Processing/
Use:
N/
A
Hypothesis
tested:
To
determine
the
levels
of
PFOS
in
the
serum
of
American
Red
Cross
blood
banks
in
6
regions
of
the
U.
S.

Study
period:
2000
Setting:
N/
A
Total
population:
6
ARC
blood
banks
in
various
geographic
regions
in
the
US
provided
645
serum
samples
from
adult
donors.
The
6
regions
included:
Los
Angeles,
CA;
Minneapolis/
St.
Paul,
MN;
Charlotte,
NC;
Boston,
MA;
Portland,
OR,
and
Hagerstown,
MD.

Subject
selection
criteria:
Unknown
Total
#
of
subjects
in
study:
645
donors,
age
20­
69
years
Comparison
population:
N/
A
Participation
rate:
N/
A
Subject
description:
The
only
demographic
factors
known
were
age,
gender,
and
location.

Health
effects
studied:
Levels
of
7
fluorochemicals
in
human
blood
serum,
including
PFOS.
The
other
chemicals
were:
PFOA,
PFOSAA,
M570,
M556,
PFOSA,
PFHS.

Data
collection
methods:
Each
blood
bank
was
requested
to
provide
approximately
10
samples
per
10­
year
age
intervals
(
20­
29,
30­
39,
etc.)
for
each
sex.

Details
on
data
collection:
Blood
sera
samples
were
analyzed
using
high­
pressure
liquid
chromatography/
electrospray
tandem
mass
spectrometry
(
HPLC/
ESMSMS).
24
samples
were
split
and
analyzed
to
provide
an
estimate
of
the
reliability
of
the
analyses
conducted.
The
analytical
lab
was
blind
to
the
identity
of
these
split
samples.
These
analyses
were
performed
concurrently
with
all
other
analyses
of
the
study
to
minimize
experimental
error.

Exposure
period:
Unknown­­
PFOS
serum
levels
used
as
surrogate
for
exposure.
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Description/
delineation
of
exposure
groups/
categories:
332
male
donors,
313
female
donors.
There
were
10
or
more
subjects
for
both
males
and
females
in
each
age
category
(
20­
29,
30­
39,
etc.)
except
the
60­
69
age
group
where
there
were
fewer.

Measured
or
estimated
exposure:
N/
A
Exposure
levels:
N/
A
Statistical
methods:
A
reliability
assessment
was
done.
Geometric
means,
95%
confidence
intervals
of
the
geometric
means,
range,
interquartile
range,
raw
cumulative
90th
percentile,
and
frequency
distributions,
were
calculated.
Central
tendency
and
distribution
of
the
data
by
age,
gender,
and
location
were
presented.
Bootstrap
analysis
was
done
to
calculate
mean
serum
PFOS
values
for
each
6
locations
adjusted
for
10­
year
age
intervals,
gender,
and
their
interaction
terms.
Multivariable
regression
analyses.
A
reliability
assessment
was
also
done
for
the
chemical
analyses.

Other
methodological
information:
N/
A
RESULTS
Describe
results:
The
results
of
the
reliability
analysis
indicate
that
there
was
a
strong
correlation
between
the
split
samples
(
r
=
.9)
for
PFOS
(
as
well
as
PFOA
and
PFHS).

PFOS
is
the
only
chemical
that
met
the
criteria
for
a
log­
normal
distribution
based
on
the
Shapiro­
Wilk
test.

The
geometric
mean
serum
PFOS
level
for
all
locations
and
gender
was
34.9
ppb
(
95%
CI,
33.3­
36.5
ppb).
The
range
was
<
LLOQ
(
4.3
ppb)
to
1656
ppb.
Males
had
significantly
higher
(
p
<
.05)
geometric
mean
PFOS
levels
than
females.
The
geometric
mean
for
all
males
was
37.8
ppb
(
95%
CI,
35.5­
40.3)
and
was
31.3
ppb
for
all
females
(
95%
CI,
30.0
 
34.3).
Age
was
not
an
important
predictor
of
adult
serum
fluorochemical
concentrations.

When
stratified
by
geographic
location,
the
highest
geometric
mean
for
PFOS
was
in
the
samples
from
Charlotte,
NC
(
51.5
ppb,
range:
19.3
 
166.0)
and
the
lowest
from
Boston
(
28.0
ppb,
range:
4.3
 
87.2).
The
other
PFOS
geometric
means
and
ranges
by
location
were:
Los
Angeles,
40.4
ppb
(
6.6
 
205.0);
Minneapolis/
St.
Paul,
33.1
ppb
(
7.7
 
207.0);
Portland,
27.0
ppb
(
6.0
 
1656);
Hagerstown,
35.3
ppb
(
7.6
 
226.0).
The
cumulative
90%
were
highest
for
Charlotte
(
105.3),
Minneapolis/
St.
Paul
(
71.7),
Los
Angeles
(
70.1),
and
Hagerstown
(
69.8).
Portland
and
Boston
were
much
lower
(
49.4
and
48.7,
respectively).

The
results
from
a
bootstrap
analysis,
done
to
calculate
mean
serum
PFOS
values
for
each
6
locations
adjusted
for
10­
year
age
intervals,
gender,
and
their
interaction
terms,
resulted
in
similar
means
for
Boston
(
29.0
ppb),
Los
Angeles
(
35.0
ppb),
Minneapolis/
St.
Paul
(
34.8
ppb),
and
Hagerstown
(
34.9
ppb).
However
the
mean
for
Charlotte
was
much
lower
(
39.0
ppb)
and
the
mean
for
Portland
was
slightly
higher
(
32.8
ppb).

Bootstrap
analyses
for
PFOS
calculated
a
mean
of
the
95%
tolerance
limit
of
88.5
ppb
with
an
upper
95%
confidence
limit
of
100.0
ppb.
The
mean
of
the
99%
tolerance
limit
was
157.3
ppb
with
an
upper
95%
confidence
limit
of
207.0
ppb.
At
the
lowest
tolerance
limit
analyzed
(
90%),
the
mean
for
PFOS
was
70.7
ppb
with
an
upper
95%
confidence
limit
of
74.3
ppb.

The
highest
serum
PFOS
measurement
in
this
sample
was
1656
ppb
from
a
male
blood
donor,
67
years
old
ENV/
JM/
RD(
2002)
17/
FINAL
349
from
Portland.
The
next
highest
donor
level
was
329
ppb
from
a
male
donor,
62
years
old
also
from
Portland.
The
next
8
highest
serum
PFOS
values
(
range
139
 
226
ppb)
were
measured
in
4
females
and
4
males
representing
Charlotte
(
n=
4),
Hagerstown
(
n=
2),
Los
Angeles
(
n=
1)
and
Minneapolis/
St.
Paul
(
n=
1).

PFOS
and
PFOA
were
strongly
correlated
(
r
=
.63)
PFOS
had
a
lower
correlation
with
PFOSAA
(
r
=
.42)
and
M570
(
r
=
.20).
Both
PFOSAA
and
M570,
adjusted
for
age,
gender,
and
their
interaction,
were
significant
predictors
of
PFOS
in
a
multivariable
model.
PFOSAA
was
the
stronger
of
the
2
independent
variables.
Age
and
gender
were
not
significant
predictors
in
models
that
examined
the
significant
association
between
PFOS
and
PFOA.

Study
strengths
and
weaknesses:
Blood
donors
cannot
be
considered
representative
of
the
general
population
of
the
US.

Research
sponsors:
3M
Medical
Department,
Corporate
Occupational
Medicine
Consistency
of
results:
Mean
PFOS
levels
reported
in
this
study
are
similar
to
those
reported
in
other
pooled
blood
samples.

CONCLUSIONS
N/
A
REFERENCE
Olsen,
GW,
Burris,
JM,
Lundberg,
JK,
Hansen,
KJ,
Mandel,
JH,
Zobel,
LR.
Identification
of
fluorochemicals
in
human
sera.
I.
American
Red
Cross
adult
blood
donors.
Final
report.
February
25,
2002.
ENV/
JM/
RD(
2002)
17/
FINAL
350
EPIDEMIOLOGIC
DATA
Title:
Identification
of
Fluorochemicals
in
Human
Sera.
II.
Elderly
Participants
of
the
Adult
Changes
in
Thought
Study,
Seattle,
Washington
TEST
SUBSTANCE
Identity:
PFOS
and
6
other
fluorochemicals
Remarks:
The
results
reported
are
final.

METHOD
Study
design:
Cross­
sectional.

Manufacturing/
Processing/
Use:
N/
A
Hypothesis
tested:
To
determine
the
serum
concentrations
of
selected
fluorochemicals
in
a
sample
of
elderly
persons
to
provide
a
more
specific
understanding
of
the
distribution
of
these
compounds
in
this
age
group.

Study
period:
9/
29/
2000
Setting:
N/
A
Total
population:
238
serum
samples
from
elderly
adult
donors
from
the
Adult
Changes
in
Thought
study.

Subject
selection
criteria:
Donors
were
65­
96
years
old.
Subjects
were
identified
during
an
enrollment
phase
of
this
community­
based
prospective
cohort
study
of
dementia
and
normal
aging
conducted
collaboratively
between
the
U.
of
Washington
and
Group
Health
Cooperative
(
HMO).
Eligible
individuals
were
those
with
no
known
history
of
neuropsychiatric
disease
or
dementia.

Total
#
of
subjects
in
study:
238
Comparison
population:
N/
A
Participation
rate:
N/
A
Subject
description:
238
adults­­
118
males,
120
females.
The
mean
age
was
76
years.
Female
subjects
had
resided
in
the
Seattle
area
for
53.3
years,
males
50.2
years.

Health
effects
studied:
PFOS
serum
levels
in
blood,
as
well
as
6
other
fluorochemicals.

Data
collection
methods:
Blood
sera
samples
were
collected
using
high­
pressure
liquid
chromatography/
electrospray
tandem
mass
spectrometry
(
HPLC/
ESMSMS).

Exposure
period:
N/
A
ENV/
JM/
RD(
2002)
17/
FINAL
351
Description/
delineation
of
exposure
groups/
categories:
65­
75
years
(
n=
121),
75+
to
85
years
(
n
=
93),
85+
to
96
years
(
n
=
24).

Measured
or
estimated
exposure:
N/
A
Exposure
levels:
N/
A
Statistical
methods:
Arithmetic
means,
ranges,
geometric
means
and
95%
confidence
intervals
were
calculated.
Central
tendency
and
distribution
of
the
data
by
age,
gender,
location
and
their
respective
interaction
terms.
A
reliability
assessment
was
also
done.

Other
methodological
information:
N/
A
RESULTS
Describe
results:
The
geometric
mean
of
PFOS
for
all
samples
was
31.0
ppb
(
95%
CI,
28.8­
33.4).
The
range
was
3.4
 
175
ppb.
There
was
no
significant
(
p
<
.05)
difference
in
geometric
means
for
males
and
females.
In
simple
linear
regression
analyses,
age
was
negatively
(
p
<
.05)
associated
with
PFOS
in
men
but
not
in
women.
The
mean
of
the
95%
tolerance
limit
for
PFOS
was
84.1
ppb
with
an
upper
95%
confidence
limit
of
104.0
ppb.

Study
strengths
and
weaknesses:
These
data
are
cross­
sectional
data
used
to
determine
PFOS
levels
in
elderly.
Very
little
descriptive
information
about
the
subjects
is
available.
The
subjects
only
characterize
PFOS
levels
in
the
Seattle
region.
To
date,
they
are
the
only
data
available
characterizing
serum
PFOS
levels
in
the
elderly.

Research
sponsors:
3M
Medical
Department,
Corporate
Occupational
Medicine
Consistency
of
results:
To
date,
no
other
data
have
been
collected
on
PFOS
serum
levels
in
elderly.

CONCLUSIONS
PFOS
levels
in
these
Seattle
residents
are
similar
to
those
found
in
adults
and
children
in
the
U.
S.

REFERENCE
Olsen,
GW,
Burris,
JM,
Lundberg,
JK,
Hansen,
KJ,
Mandel,
JH,
Zobel,
LR.
February
25,
2002.
Identification
of
fluorochemicals
in
human
sera.
II.
Elderly
participants
of
the
Adult
Changes
in
Thought
study,
Seattle,
Washington.
Final
Report.
3M
Company,
Medical
Department.
ENV/
JM/
RD(
2002)
17/
FINAL
352
EPIDEMIOLOGIC
DATA
Title:
Identification
of
Fluorochemicals
in
Human
Sera.
III.
Pediatric
Participants
in
a
Group
A
Streptococci
Clinical
Trial
Investigation
TEST
SUBSTANCE
Identity:
PFOS
and
6
other
fluorochemicals
Remarks:
The
results
reported
are
final.

METHOD
Study
design:
Cross­
sectional.

Manufacturing/
Processing/
Use:
N/
A
Hypothesis
tested:
To
determine
the
serum
concentrations
of
selected
fluorochemicals
in
a
sample
of
children
to
provide
a
more
specific
understanding
of
the
distribution
of
these
compounds
in
children.

Study
period:
Child
sera
samples
were
collected
from
January
1994
to
March
1995.
The
sera
samples
were
analyzed
in
Spring
1999.

Setting:
N/
A
Total
population:
Not
reported
Subject
selection
criteria:
The
sera
samples
were
provided
to
3M
by
the
University
of
Minnesota
Department
of
Pediatrics.
They
were
obtained
from
a
large
clinical
trial
on
Group
A
streptococcal
infections
in
children.
The
children
were
residents
of
23
states
in
the
US.
These
children
presented
with
signs
and
symptoms
of
acute­
onset
pharyngitis.
All
of
the
children
had
positive
throat
cultures
at
the
initial
visit.

Total
#
of
subjects
in
study:
n
=
599
children,
age
2­
12
years
Comparison
population:
N/
A
Participation
rate:
N/
A
Subject
description:
299
male
children,
300
female
children
from
23
states
and
the
District
of
Columbia.

Health
effects
studied:
PFOS
serum
levels
in
blood,
as
well
as
6
other
fluorochemicals.

Data
collection
methods:
Blood
sera
samples
were
collected
using
high­
pressure
liquid
chromatography/
electrospray
tandem
mass
spectrometry
(
HPLC/
ESMSMS).
Sera
were
frozen
at
 
20
degrees
C
prior
to
the
request
for
analysis.

Exposure
period:
N/
A
ENV/
JM/
RD(
2002)
17/
FINAL
353
Description/
delineation
of
exposure
groups/
categories:
Blood
sera
samples
were
collected
from
children
2
 
12
years
old.

Measured
or
estimated
exposure:
N/
A
Exposure
levels:
N/
A
Statistical
methods:
Arithmetic
means,
ranges,
geometric
means
and
95%
confidence
intervals
were
calculated.
Central
tendency
and
distribution
of
the
data
by
age,
gender,
location
and
their
respective
interaction
terms.
A
reliability
assessment
was
also
done.

Other
methodological
information:
N/
A
RESULTS
Describe
results:
The
geometric
mean
of
PFOS
for
all
of
the
participants
was
37.5
ppb
(
95%
CI,
33.3­
36.5).
The
range
was
6.7
to
515.0
ppb.
Male
children
had
significantly
(
p<.
01)
higher
geometric
mean
PFOS
levels
than
females:
40.1
ppb
and
35.2
ppb,
respectively.
In
bootstrap
analyses,
the
mean
of
the
95%
tolerance
limit
for
PFOS
was
88.5
ppb
with
an
upper
95%
confidence
limit
of
97.0
ppb.
When
stratified
by
age,
the
geometric
mean
tended
to
rise
for
each
age
group
from
age
2
(
28.6
ppb)
through
age
9
(
42.8
ppb)
where
it
was
highest,
and
then
started
to
decrease
gradually
to
32.8
ppb
at
12
years.
In
simple
linear
regression
analyses,
age
was
not
significantly
(
p
<
.05)
associated
with
PFOS.
Although
the
data
were
not
provided,
a
graphical
presentation
of
log
PFOS
levels
for
each
state
by
gender
were
similar
across
the
states,
however,
it
is
difficult
to
interpret
these
data
given
the
limited
sample
size
for
each
gender/
location
subgroup.

Study
strengths
and
weaknesses:
These
data
are
cross­
sectional
data
used
to
determine
PFOS
levels
in
U.
S.
children.
Very
little
descriptive
information
about
the
subjects
is
available.
To
date,
they
are
the
only
data
available
characterizing
serum
PFOS
levels
in
children.

Research
sponsors:
3M
Medical
Department,
Corporate
Occupational
Medicine
Consistency
of
results:
To
date,
no
other
data
have
been
collected
on
PFOS
serum
levels
in
children.

CONCLUSIONS
N/
A
REFERENCE
Olsen,
GW,
Burris,
JM,
Lundberg,
JK,
Hansen,
KJ,
Mandel,
JH,
Zobel,
LR.
Identification
of
fluorochemicals
in
sera
of
children
in
the
United
States.
Interim
Report.
June
25,
2001.
ENV/
JM/
RD(
2002)
17/
FINAL
354
EPIDEMIOLOGIC
DATA
Title:
A
Cross­
sectional
analysis
of
serum
perfluorooctanesulfonate
(
PFOS)
and
Perfluorooctanoate
(
PFOA)
in
relation
to
clinical
chemistry,
thyroid
hormone,
hematology,
and
urinalysis
results
from
male
and
female
employee
participants
of
the
2000
Antwerp
and
Decatur
fluorochemical
medical
surveillance
program
TEST
SUBSTANCE
Identity:
PFOS,
PFOA
Remarks:

METHOD
Study
design:
cross­
sectional
Manufacturing/
Processing/
Use:
Facilities
in
Decatur,
Alabama
and
Antwerp,
Belgium
which
manufacture
perfluorooctanesulfonyl
fluoride
products.
These
fluorochemicals
can
metabolize
in
the
body
to
PFOS.

Hypothesis
tested:
To
provide
an
aggregate
analysis
of
the
hematology,
clinical
chemistries,
and
hormonal
parameters
of
volunteer
employees
in
relation
to
serum
PFOS
and
PFOA
levels
as
measured
in
the
medical
surveillance
examinations
of
Antwerp
and
Decatur
employees
in
2000.

Study
period:
March
1,
2000.
End
date
was
not
reported.

Setting:
Occupational.
3M
plants
located
in
Antwerp,
Belgium
and
Decatur,
Alabama.

Total
population:
340
Antwerp
employees
and
500
Decatur
employees
working
in
the
chemical
plant
area
were
eligible
for
inclusion
in
the
surveillance.

Subject
selection
criteria:
Voluntary
participation
in
medical
surveillance
program
in
Y2000.

Total
#
of
subjects
in
study:
255
Antwerp
employees
(
206
male
and
49
female)
and
263
Decatur
employees
(
215
male
and
48
female).

Comparison
population:
N/
A
Participation
rate:
75%
of
employees
at
the
Antwerp
plant
and
50%
of
the
employees
at
the
Decatur
plant
who
were
eligible
participated.
73%
of
the
participating
Antwerp
male
employees
and
75%
of
the
Decatur
employees
were
engaged
in
production
activities.
Only
12%
of
the
participating
Antwerp
female
employees
were
engaged
in
production
activities
compared
to
63%
of
the
Decatur
female
employees.

Subject
description:
Male
Antwerp
employees
had
lower
PFOS
and
PFOA
levels,
were
significantly
younger
than
Decatur
male
employees,
had
lower
BMIs,
worked
fewer
years,
had
higher
self­
reported
daily
consumption
of
alcohol,
had
lower
mean
alkaline
phosphatase,
GGT,
AST,
ALT
and
triglyceride
values
and
higher
total
bilirubin
and
HDL
values.
Comparable
results
were
observed
for
Antwerp
female
employees
vs.
Decatur
females.

Health
effects
studied:
To
determine
if
there
were
differences
in
the
following
parameters
based
on
ENV/
JM/
RD(
2002)
17/
FINAL
355
PFOS/
PFOA
levels:
hematology
(
hematocrit,
hemoglobin,
RBCs,
WBCs,
platelet
count),
clinical
chemistries
(
alkaline
phosphatase,
gamma
glutamyl
transferase,
aspartate
aminotransferase,
alanine
aminotransferase,
total
and
direct
bilirubin,
blood
urea
nitrogen,
creatinine,
glucose,
cholesterol,
low
density
lipoproteins,
high
density
lipoproteins,
and
triglycerides),
and
thyroid
hormones
(
thyroid
stimulating
hormone,
serum
thyroxine,
free
thyroxine,
serum
triiodothyronine,
thyroid
hormone
binding
ratio,
and
free
thyroxine).

Data
collection
methods:
Medical
questionnaire,
work
history
questionnaire,
blood
sera
samples,
measurements
of
height,
weight,
and
blood
pressure,
urinalysis
(
Decatur
only),
and
standard
clinical
chemistry
and
hematology
tests,
thyroid
hormone
measurement,
and
pulmonary
function
tests.
Values
used
for
reference
ranges
were
not
provided.

Details
on
data
collection:
The
site­
specific
work
history
questionnaire
was
administered
to
all
participants.
The
data
were
self­
reported.
Questionnaire
content,
design,
administration,
etc.
were
not
provided
in
this
report.
Data
on
blood
collection
(
amount,
etc.)
not
provided.
Urinalysis
was
only
assessed
for
Decatur
employees
via
standard
urine
microstick
analysis
which
tested
for
urine
glucose,
albumin,
and
RBCs.

TSH,
free
T4
and
T3
were
determined
by
immunochemiluminometric
assay.
T4
and
THBR
were
determined
by
a
cloned
enzyme
donor
immunoassay.
FTI
was
calculated
by
multiplying
T4
and
THBR.

Sera
samples
were
extracted
using
an
ion­
pairing
extraction
procedure.
In
addition
to
PFOA
and
PFOS,
the
extracts
were
also
analyzed
for
PFHS,
PFOSAA,
PFOSA,
and
M556
(
perfluorooctanesulfonamidoacetate)
using
high­
pressure
liquid
chromatography
electrospray
tandem
mass
spectrometry
and
evaluated
versus
an
extracted
curve
from
a
human
serum
matrix.
All
serum
values
for
PFOS
and
PFOA
were
above
the
LLOQ.

Exposure
period:
Unknown.
PFOS/
PFOA
serum
levels
indicate
exposure.

Description/
delineation
of
exposure
groups/
categories:
Workers
were
stratified
by
plant
location
as
well
as
by
serum
PFOS
distribution,
production
status
(
production
vs.
non­
production
workers),
and
gender.

Mean
serum
PFOS
levels
for
all
employees
participating
in
this
study
at
Antwerp
(
n
=
206)
and
Decatur
(
n
=
215)
were
0.96
and
1.40
ppm,
respectively.
Levels
among
production
employees
were
higher.
At
Antwerp,
the
mean
PFOS
level
of
male
production
employees
was
1.16
ppm
and
1.63
ppm
at
Decatur.

Mean
PFOA
levels
for
all
employees
were
1.03
and
1.90
ppm
at
Antwerp
and
Decatur,
respectively.
Levels
among
production
employees
were
higher.
At
Antwerp,
the
mean
PFOA
level
of
male
production
employees
was
1.28
ppm
and
2.34
ppm
at
Decatur.

Measured
or
estimated
exposure:
Serum
PFOS
and
PFOA
levels
were
used
to
estimate
exposure.

Statistical
methods:
Descriptive
simple
and
stratified
analyses,
Pearson
correlation
coefficients,
analysis
of
variance,
and
multivariable
regression
were
used
to
evaluate
associations
between
PFOS
and
PFOA
and
each
hematological
and
clinical
chemistry
test
and
thyroid
hormone
assay.
For
stratified
analyses,
employees
were
divided
into
quartiles
of
their
serum
PFOS
distribution.
Potential
confounding
factors
considered
in
the
analyses
included:
age,
BMI,
alcohol
consumption,
cigarette
use,
years
worked
at
either
plant,
and
type
of
job.

Multivariable
regression
models
were
fitted
with
PFOS/
PFOA
analyzed
as
continuous
variables.
Natural
ENV/
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2002)
17/
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356
log
transformations
of
the
dependent
variables
were
performed,
when
necessary,
to
normalize
variables
and
to
enhance
model
fit.
SAS
was
used
to
analyze
the
data.

Other
methodological
information:

RESULTS
Describe
results:
Antwerp
and
Decatur
employees
were
different
in
several
ways
(
see
"
Subject
description"
above).
Therefore,
univariate
analyses
were
initially
stratified
by
location
and
then
those
analyses
were
stratified
by
gender
and
production
status.
They
were
placed
into
quartiles
depending
on
production
status.
Therefore,
the
PFOS
values
in
the
quartiles
are
different
for
male
production,
non­
production,
and
female
employees.

Antwerp
The
mean
PFOS
level
for
all
employees
at
this
plant
was
0.96
ppm
(
range
0.04
­
6.24
ppm).
When
stratified
by
production
status,
the
mean
was
1.16
ppm
for
production
employees
and
0.42
ppm
for
nonproduction
employees.
The
mean
for
female
employees
was
0.13
ppm.

Male
production
employees
were
placed
into
the
following
quartiles
based
on
PFOS
levels:
Q1
(
mean,
0.29
ppm;
range,
0.04
 
0.41
ppm),
Q2
(
mean,
0.58
ppm;
range
0.41
 
0.78
ppm),
Q3
(
mean
1.18
ppm;
range
0.79
 
1.66
ppm),
Q4
(
mean,
2.61
ppm;
range,
1.67
 
6.24
ppm).

In
male
production
employees
(
n
=
150),
the
highest
quartile
mean
serum
PFOS
level
was
2.61
ppm
(
range
1.76
 
6.24
ppm)
and
the
lowest
was
0.29
ppm
(
range
0.04
 
0.41
ppm).
Production
employees
in
the
highest
quartile
were
significantly
(
p
<
.05)
older
and
worked
more
years
at
Antwerp
than
employees
in
the
lowest
quartile.
The
only
difference
in
clinical
chemistries
for
production
workers
was
in
BUN.
When
compared
by
quartile
of
serum
PFOS
distribution,
no
significant
(
p
<.
05)
differences
among
male
production
employees
were
observed
for
thyroid
(
TSH,
T4,
free
T4,
T3,
THBR,
FTI)
or
for
hematology
(
HCT,
HGB,
RBC,
WBC,
platelets).
The
same
held
true
for
non­
production
employees
(
n
=
56).

For
all
female
employees,
BUN
was
significantly
different
(
higher)
between
1st
quartile
and
3rd
and
4th
quartile.
Thyroid
and
hematology
results
were
not
significantly
different
between
any
of
the
quartiles
for
females.

Decatur
The
mean
PFOS
level
for
all
employees
at
this
plant
was
1.40
ppm
(
range
0.11
 
10.06
ppm).
When
stratified
by
production
status,
the
mean
was
1.63
ppm
for
production
employees
and
0.73
ppm
for
nonproduction
employees.
The
mean
for
female
employees
was
0.93
ppm.
75%
of
male
employees
worked
in
production
jobs
(
n
=
161)
and
63%
of
female
employees
worked
in
production
jobs.

Male
production
employees
were
placed
into
the
following
quartiles
based
on
PFOS
levels:
Q1
(
mean,
0.55
ppm;
range,
0.11
 
0.75
ppm),
Q2
(
mean,
1.01
ppm;
range
0.76
 
1.30
ppm),
Q3
(
mean
1.74
ppm;
range
1.32
 
2.29
ppm),
Q4
(
mean,
3.22
ppm;
range,
2.31
 
10.06
ppm).

When
male
production
workers
were
placed
into
quartiles,
the
only
significant
(
p
<
.05)
difference
between
the
quartiles
was
in
ALT
(
highest
quartile
different
from
all
3
others).
No
significant
differences
between
quartiles
were
observed
for
thyroid
(
TSH,
T4,
free
T4,
T3,
THBR,
FTI),
hematology
(
HCT,
HGB,
RBC,
WBC,
platelets),
or
urinalysis
(
albumin,
blood,
sugar).
ENV/
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For
female
employees
(
both
production
and
non­
production
employees),
there
were
no
significant
differences
among
quartiles
for
demographics,
thyroid,
hematology,
or
urinalysis
except
for
mean
platelet
count
where
the
third
quartile
was
significantly
lower
than
the
1st
quartile,
but
the
4th
was
not.

When
results
were
analyzed
by
number
of
employees
who
had
values
above
the
reference
range
for
hepatic
clinical
chemistry
tests
and
liver
enzyme
and
bilirubin
tests,
there
was
a
higher
percentage
of
male
Decatur
production
workers
in
the
highest
PFOS
quartile
for
ALT,
GGT,
and
total
liver
panel
than
the
other
quartiles.
Most
notable
were
the
results
for
ALT
where
8%
of
employees
in
the
lowest
exposure
group
(
Q1)
and
28%
in
the
highest
exposure
group
(
Q4)
had
values
above
the
reference
range,
while
the
percentages
for
total
liver
panel
(
which
includes
alkaline
phosphatase,
AST,
ALT,
GGT,
and
total
and
direct
bilirubin)
were
18%
and
35%,
respectively.
This
trend
was
not
evident
in
Decatur
non­
production
employees
(
although
the
n
was
only
54),
in
Decatur
females,
or
in
any
of
the
Antwerp
employees.
However,
each
sub­
population
had
a
different
serum
PFOS
quartile
distribution.
Therefore,
they
cannot
be
directly
compared.

Analyses
combining
employees
from
both
plants
When
clinical
chemistry
results
of
all
male
employees
from
both
plants
were
combined
(
both
production
and
non­
production)
(
n
=
421)
and
placed
into
quartiles
(
n
=
105
per
quartile),
mean
values
for
triglycerides,
alkaline
phosphatase,
total
bilirubin,
and
ALT
were
significantly
(
p
<
.05)
higher
in
the
4th
quartile
(
mean
PFOS
level
2.69,
range
1.69
 
10.06
ppm)
than
in
the
first
(
mean
PFOS
level
0.27
ppm,
range
0.04
 
0.42
ppm).
It
should
be
noted
that
the
number
of
Antwerp
production
employees
were
evenly
distributed
among
the
quartiles
while
this
was
not
the
case
for
Decatur
employees.
The
highest
number
of
Decatur
employees
was
in
the
4th
quartile.
In
addition,
overall
the
employees
in
the
1st
quartile
were
slightly
younger,
had
a
lower
BMI,
and
worked
fewer
years
than
employees
in
the
other
quartiles.

Thyroid
results
for
this
same
group
indicated
that
T3
was
significantly
higher
(
p
<
.05)
and
THBR
was
significantly
lower
(
p
<
.05)
in
Q4
than
Q1.
In
female
employees
combined
for
both
plants
(
n
=
97),
alkaline
phosphatase
and
GGT
were
significantly
higher
(
p
<
.05)
and
total
bilirubin
significantly
lower
in
Q4
than
in
Q1.
Most
of
the
Decatur
female
employees
worked
in
production
jobs
while
most
of
the
Antwerp
females
worked
in
non­
production
jobs.
Therefore,
Q4
was
92%
female
production
workers
and
all
of
them
worked
at
the
Decatur
plant.

The
combined
plant
data
were
analyzed
for
employees
who
had
values
above
the
reference
range
for
alkaline
phosphatase,
AST,
ALT,
GGT,
and
total
liver
panel.
For
male
employees
for
all
of
these
measures,
the
levels
increased
from
Q1
to
Q4.
In
Q1,
4%
of
the
employees
had
values
above
the
reference
range
for
ALT
and
6%
for
GGT,
while
12%
was
reported
for
Q4
for
both
of
these
tests.
For
total
liver
panel,
14%
of
the
employees
had
values
above
the
reference
range
in
Q1
as
compared
to
23%
in
Q4.
The
numbers
of
female
employees
with
values
above
the
reference
range
was
very
small
(
n
=
8).

The
above
data
were
not
adjusted
for
potential
confounders;
therefore,
multivariable
regression
analyses
were
conducted.
There
was
a
positive
significant
(
p
=
.04)
association
between
PFOS
and
cholesterol
and
also
a
positive
significant
(
p
=
.05)
association
between
PFOA
and
cholesterol.
When
both
PFOS
and
PFOA
were
included
in
the
model,
neither
were
statistically
significant
at
p=.
05.
PFOS
was
not
significant
with
HDL
although
PFOA
was
negatively
associated
with
HDL
(
p
=
.04).
Triglycerides
were
positively
associated
with
PFOS
(
p
=
.01)
and
PFOA
(
p
=
.002).
When
both
were
left
in
the
model,
PFOA
remained
significant
(
p
=
.02).
Total
organic
fluorine
(
TOF)
was
highly
significant
for
triglycerides
(
p
=
.0009).
No
significant
associations
were
observed
with
PFOS,
PFOA,
or
TOF
in
relation
to
alkaline
phosphatase,
GGT,
AST
or
total
bilirubin.
A
significant
(
p
=
.02)
positive
association
was
observed
for
TOF
and
ALT.
A
positive
significant
(
p
=
.04)
association
between
T3
and
PFOS
was
observed.
Plant
location
was
highly
significant
(
p
<
.0001)
in
the
model.
BMI,
cigarettes/
day,
alcohol/
day
were
also
significant.
[
In
the
ENV/
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2002)
17/
FINAL
358
univariate
analyses,
Antwerp
employees
had
higher
mean
T3
levels
than
Decatur
employees
overall.
However,
for
each
plant
(
individually)
T3
values
increased
by
quartile
as
PFOS
serum
levels
increased,
although
the
differences
were
not
statistically
significant.]
THBR,
as
well
as
the
other
thyroid
hormones,
were
not
significant
in
the
regression
analyses.
Most
of
the
thyroid
hormones,
including
TSH,
T4,
THBR,
and
FTI,
were
not
significantly
associated
with
PFOS,
PFOA,
or
TOF.
However,
PFOS,
PFOA
and
TOF
were
positively
associated
(
p
=
.04,
.01,
and
.004,
respectively)
with
T3.

Study
strengths
and
weaknesses:
Cross­
sectional
design,
voluntary
participation,
the
Decatur
and
Antwerp
populations
were
significantly
different
in
certain
demographic
and
clinical
chemistry
results
as
well
as
in
PFOS
serum
levels,
PFOS
serum
levels
are
declining
thereby
making
it
harder
to
detect
an
effect
if
one
is
present,
plant
populations
cannot
be
compared
because
quartiles
are
different
for
each
subgroup,
only
one
measurement
at
a
certain
point
in
time
was
collected
for
each
test,
other
perfluorinated
chemicals,
such
as
PFOA,
are
present
in
the
plants.

Research
sponsors:
3M
Consistency
of
results:
These
results
are
somewhat
consistent
with
those
of
the
1995
and
1997
crosssectional
medical
surveillance
data,
in
that
all
of
them
conclude
that
there
were
no
significant
abnormalities
in
hematological
and
clinical
chemistry
parameters
of
the
Antwerp
or
Decatur
workers.
No
decline
in
cholesterol
levels
was
observed
as
PFOS
serum
levels
increased.
The
hormone
data
collected
in
1995
was
different
from
that
collected
in
2000
and
therefore
cannot
be
compared.
A
longitudinal
analysis
of
these
data
did
not
reveal
any
significant
changes
in
hepatic
or
lipid
clinical
chemistry
values;
however,
there
were
many
limitations
to
the
study.

CONCLUSIONS
The
authors
concluded
that
these
data
continue
to
suggest
that
Antwerp
and
Decatur
fluorochemical
production
and
non­
production
employees
do
not
have
significant
changes
in
serum
cholesterol,
lipoproteins,
or
hepatic
enzymes
that
are
consistent
with
toxicological
findings
in
laboratory
animals.

REFERENCE
Olsen
GW,
Burlew
MM,
Burris
JM,
Mandel
JH.
October
11,
2001.
A
cross­
sectional
analysis
of
serum
perfluorooctanesulfonate
(
PFOS)
and
perfluorooctanoate
(
PFOA)
in
relation
to
clinical
chemistry,
thyroid
hormone,
hematology
and
urinalysis
results
from
male
and
female
employee
participants
of
the
2000
Antwerp
and
Decatur
fluorochemical
medical
surveillance
program.
Final
report.
3M
Medical
Department.
ENV/
JM/
RD(
2002)
17/
FINAL
359
EPIDEMIOLOGIC
DATA
Title:
A
Longitudinal
Analysis
of
Serum
Perfluorooctanesulfonate
(
PFOS)
and
Perfluorooctanoate
(
PFOA)
Levels
in
Relation
to
Lipid
and
Hepatic
Clinical
Chemistry
Test
Results
from
Male
Employee
Participants
of
the
1994/
95,
1997,
and
2000
Fluorochemical
Medical
Surveillance
Program
TEST
SUBSTANCE
Identity:
PFOS,
PFOA
Remarks:

METHOD
Study
design:
longitudinal
Manufacturing/
Processing/
Use:
3M
Decatur,
Alabama
plant
and
Antwerp,
Belgium
plant.

Hypothesis
tested:
To
determine
whether
occupational
exposure
to
fluorochemicals
over
time
is
related
to
changes
in
clinical
chemistry
and
lipid
results
in
employees
of
2
3M
facilities.

Study
period:
There
were
3
time
periods
during
which
medical
surveillance
took
place
at
the
plants
 
1994/
95,
1997,
and
2000.

Setting:
3M
plants
in
Decatur,
Alabama
and
Antwerp,
Belgium.

Total
population:
175
male
employees
participated
in
2000
and
at
least
one
of
the
other
sampling
periods.

Subject
selection
criteria:
Employees
participated
voluntarily.

Comparison
population:
n/
a
Participation
rate:
106/
175
(
61%)
participated
in
1994/
95,
110/
175
(
63%)
participated
in
1997,
and
175
participated
in
2000.
24%
participated
in
all
3
sampling
periods
(
n
=
41,
Antwerp
20,
Decatur
20),
37%
in
1994/
95
and
2000
(
n
=
65,
Antwerp
45,
Decatur
20),
and
39%
(
n
=
69,
Antwerp
34,
Decatur
35)
in
1997
and
2000.

Subject
description:
Male
employees
volunteered
to
participate
in
biomonitoring
offered
at
the
plants.
In
general,
Antwerp
male
employees
were
significantly
younger,
had
lower
BMIs
and
a
higher
daily
consumption
of
alcohol
reported
than
Decatur
male
employees.
Antwerp
male
employees
also
had
lower
mean
alkaline
phosphatase
and
triglyceride
values
and
higher
total
bilirubin
and
HDL
values
than
the
Decatur
male
employees.

Health
effects
studied:
To
determine
whether
workers'
lipid
and
hepatic
clinical
chemistry
results
are
affected
by
PFOS
and
PFOA
levels.

Data
collection
methods:
Clinical
chemistries
and
hematology
collected
 
cholesterol
(
mg/
dl),
high
density
lipoproteins
(
HDL,
mg/
dl),
triglycerides
(
mg/
dl),
alkaline
phosphatase
(
IU/
L),
gamma
glutamyl
transferase
(
GGT,
IU/
L),
aspartate
aminotransferase
(
AST,
IU/
L),
alanine
aminotransferase
(
ALT,
IU/
L),
total
and
direct
bilirubin
(
mg/
dl).
Demographic
data
collected
via
questionnaire.
ENV/
JM/
RD(
2002)
17/
FINAL
360
Details
on
data
collection:
Details
on
data
collection
methods
including
questionnaire
content,
design,
administration,
etc.
and
blood
collection
methods
were
not
provided.

PFOS
and
PFOA
methods
of
analysis
differed
slightly
each
year.
In
1994/
95,
the
method
used
tetrabutylammonium
to
ion­
pair
with
PFOS
and
PFOA
in
the
serum.
The
ion­
pairs
were
then
extracted
with
ethyl
acetate
and
the
abstraction
product
was
then
analyzed
using
high­
performance
liquid
chromatograph­
thermospray
mass
spectrometry.
In
1997,
the
serum
samples
were
analyzed
by
liquid
chromatorgraphy/
mass
spectrometry,
using
selected
ion
monitoring
in
the
negative­
ion
mode.
In
2000,
sera
samples
were
extracted
using
an
ion­
pairing
extraction
procedure.
High­
performance
liquid
chromatography/
electrospray
tandem
mass
spectrometry
was
used.
The
samples
were
evaluated
versus
an
extracted
curve
from
a
human
serum
matrix.

Exposure
period:
Unknown.
PFOS,
PFOA
levels
measured
in
blood
serum.

Description/
delineation
of
exposure
groups/
categories:
The
groups
of
employees
were
broken
into
subpopulations
A,
B,
and
C.
A
was
comprised
of
the
employees
who
participated
in
all
3
years
of
surveillance,
B
contained
those
employees
who
participated
in
1994/
95
and
2000,
and
C
contained
those
who
participated
in
1997
and
2000.

Measured
or
estimated
exposure:
PFOS
and
PFOA
levels
were
measured
in
workers'
blood
serum.
No
ambient
exposure
data
are
available.

Exposure
levels:
Mean
PFOS
levels
Antwerp
Decatur
1994/
95
1.87
ppm
2.62
ppm
1997
1.42
ppm
1.85
ppm
2000
1.16
ppm
1.67
ppm
Mean
PFOA
levels
Antwerp
Decatur
1994/
95
1.08
ppm
1.90
ppm
1997
1.54
ppm
1.41
ppm
2000
1.43
ppm
1.83
ppm
Statistical
methods:
repeated
measures
incorporating
the
random
subject
effect
fitted
to
a
mixed
model
using
SAS.
Restricted
maximum
likelihood
estimates
of
variance
parameters
were
computed.
Adjusted
regression
models
were
built
by
introducing
all
covariates
and
testing
the
covariance
structure.
Covariates
included
in
the
model
were
age,
BMI,
number
of
alcoholic
drinks
per
day,
and
cigarettes
smoked
per
day.

Other
methodological
information:
A
total
of
175
male
employees
(
100
Antwerp
and
75
Decatur)
who
participated
in
the
2000
surveillance
year
also
participated
in
at
least
one
previous
fluorochemical
medical
surveillance
exam
since
1994/
95.
Therefore,
this
provided
an
opportunity
to
undertake
a
longitudinal
assessment.
ENV/
JM/
RD(
2002)
17/
FINAL
361
RESULTS
PFOS
results
When
mean
serum
PFOS
levels
were
compared
by
surveillance
year,
PFOS
levels
have
been
decreasing
in
the
participants
in
medical
surveillance
in
both
plants.
When
the
data
were
analyzed
by
the
3
subcohorts
(
those
who
participated
in
2
or
more
medical
exams
between
1995
and
2000),
Antwerp
and
Decatur
employees
in
each
of
the
3
subcohorts
had
lower
mean
serum
PFOS
levels
in
2000
than
at
their
year
of
entry.

When
analyzed
using
mixed
model
multivariable
regression
and
combining
Antwerp
and
Decatur
employees,
there
was
no
association
between
PFOS
and
serum
cholesterol
or
triglycerides
in
male
participants
over
time.
There
were
also
no
significant
associations
between
PFOS
and
changes
over
time
in
HDL,
alkaline
phosphatase,
GGT,
AST,
ALT,
total
bilirubin,
and
direct
bilirubin.

PFOA
results
When
mean
serum
PFOA
levels
were
compared
by
surveillance
year,
PFOA
levels
in
the
employees
participating
in
medical
surveillance
at
the
Antwerp
plant
increased
between
1994/
95
and
1997
and
then
decreased
slightly
between
1997
and
2000.
At
the
Decatur
plant,
PFOA
serum
levels
decreased
between
1994/
95
and
1997
and
then
increased
between
1997
and
2000.
When
the
data
were
analyzed
by
plant
and
the
3
subcohorts
(
those
who
participated
in
2
or
more
medical
exams
between
1995
and
2000),
there
were
no
consistent
changes
across
subcohorts
at
the
Antwerp
plant.
However,
among
the
3
Decatur
subcohorts,
mean
PFOA
levels
tended
to
increase.

When
analyzed
using
mixed
model
multivariable
regression
and
combining
Antwerp
and
Decatur
employees,
there
was
a
statistically
significant
positive
association
between
PFOA
and
serum
cholesterol
(
p
=
.0008)
and
triglycerides
(
p
=
.0002)
over
time.
When
analyzed
by
plant
and
also
by
subcohort,
these
associations
were
limited
to
the
Antwerp
employees
(
p
=
.005)
and,
in
particular,
the
21
Antwerp
employees
who
participated
in
all
3
surveillance
years
(
p
=
.001).
However,
the
association
between
PFOA
and
triglycerides
was
also
statistically
significant
(
p
=
.02)
for
subgroup
B
(
employees
who
participated
in
biomonitoring
in
1994/
95
and
2000).
There
was
not
a
significant
association
between
PFOA
and
triglycerides
among
Decatur
workers.

There
were
no
significant
associations
between
PFOA
and
changes
over
time
in
HDL,
alkaline
phosphatase,
GGT,
AST,
ALT,
total
bilirubin,
and
direct
bilirubin.

Total
Organic
Fluorine
(
TOF)
results
When
analyzed
using
mixed
model
multivariable
regression
and
combining
Antwerp
and
Decatur
employees,
there
was
a
statistically
significant
positive
association
between
TOF
and
serum
cholesterol
(
p
=
.007)
and
triglycerides
(
p
=
.008)
over
time.
However,
the
interaction
term
with
time
(
years)
was
not
significant.
This
association
was
more
consistent
for
Antwerp
employees
than
Decatur
employees.

Study
strengths
and
weaknesses:
Study
limitations
include
the
following:
1.
A
very
small
number
of
employees
participated
in
all
3
study
periods
(
only
24%,
n
=
41)
2.
different
labs
were
used
each
year
for
analysis
and
different
analytical
techniques
for
PFOS
3.
could
not
analyze
female
employees
due
to
small
numbers
4.
PFOS
levels
in
employees
are
decreasing
over
time
and
are
below
those
levels
causing
effects
in
laboratory
animals
5.
serum
PFOS
levels
were
approximately
0.5
ppm
lower
in
Antwerp
employees
than
Decatur
6.
more
Antwerp
employees
than
Decatur
participating
in
this
study
(
57%
vs.
43%)
7.
PFOA
levels
fluctuating
(
direction
depending
on
the
plant)
8.
there
are
several
consistent
differences
between
the
Antwerp
and
Decatur
male
populations
(
eg.,
ENV/
JM/
RD(
2002)
17/
FINAL
362
statistically
significant
differences
in
BMI,
age,
consumption
of
alcohol,
and
differences
in
chemistry
profiles)
9.
low
levels
of
PFOS
and
PFOA
measured
in
each
program
year
among
these
employees
as
compared
with
those
that
cause
effects
in
laboratory
animals
10.
blood
sampling
was
conducted
only
once
per
sampling
period
at
a
certain
point
in
time.

Research
sponsors:
3M
Consistency
of
results:
This
is
the
first
longitudinal
analysis
of
the
surveillance
data
and
will
probably
be
the
last
since
PFOS
is
being
phased
out.

The
positive
association
between
PFOA
and
serum
cholesterol
and
triglycerides
is
not
consistent
with
the
hypolipidemia
effect
observed
in
rodents
(
and
not
observed
in
primates).
In
addition,
this
effect
has
not
been
observed
at
3M's
Cottage
Grove
facility
where
PFOA
serum
levels
in
workers
are
much
higher
than
at
the
Decatur
or
Antwerp
plant.

CONCLUSIONS
A
longitudinal
analysis
over
a
six­
year
period
of
175
Antwerp
and
Decatur
male
employees
did
not
show
significant
changes,
consistent
with
toxicological
data,
of
lipid
or
hepatic
clinical
chemistry
values
associated
with
PFOS.
A
positive
statistically
significant
association
was
observed
between
PFOA
and
cholesterol
and
triglycerides.
When
analyzed
by
plant
and
also
by
subcohort,
these
associations
were
limited
to
the
Antwerp
employees
and,
in
particular,
the
21
Antwerp
employees
who
participated
in
all
3
surveillance
years.

REFERENCE
Olsen,
G.
W.,
Burlew,
M.
M,
Burris,
J.
M.,
Mandel,
J.
H.
A
Longitudinal
Analysis
of
Serum
Perfluorooctanesulfonate
(
PFOS)
and
Perfluorooctanoate
(
PFOA)
Levels
in
Relation
to
Lipid
and
Hepatic
Clinical
Chemistry
Test
Results
from
Male
Employee
Participants
of
the
1994/
95,
1997,
and
2000
Fluorochemical
Medical
Surveillance
Program.
3M
Final
Report.
October
11,
2001.