Document ID: EPA-HQ-OW-2004-0001-0062
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2005-05-13T04:00Z

1
SCIENCE
AND
TECHNOLOGY
GROUP
c/
o
USEPA
AWBERC
Laboratory
26
West
Martin
Luther
King
Drive
Cincinnati,
Ohio
45219
MEMORANDUM
DATE:
November
30,
2004
TO:
David
Munch,
U.
S.
EPA,
Project
Officer
FROM:
Barry
V.
Pepich,
Ph.
D.,
Program
Manager,
Shaw
Environmental,
Inc.

SUBJECT:
Research
Summary
for
EPA
Method
527.0,
Final
Version
DOC.
NO.:
TSC­
02­
0380
WGF
NO.:
004
Attached
please
find
revised
the
research
summary
for
EPA
Method
527.0.
2
Table
of
Contents
1.
Target
Compound
Selection
and
Instrument
Optimization
3
1.1.
Instrument
Configuration
1.2.
Target
Compound
List
Evaluation
2.
Extraction
Optimization
4
2.1
Solid
Phase
Evaluations
3.
Preservation
6
3.1
pH
and
Antimicrobial
Studies
3.2
Dechlorination
3.3
Degradation/
Hydrolysis
Studies
4.
Ruggedness
14
4.2
Syringe
Issues
4.2
BDE
Precision
5.
Second
Source
Standard
Evaluation
16
5.1
Individual
Standard
Analysis
5.2
Thiazopyr
and
Ethofenprox
6.
Precision
and
Accuracy
17
7.
LCMRL
and
DL
Determination
19
7.1
DL
Calculation
7.2
LCMRL
Determination
8.
Storage
Stability
20
9.
Evaluation
of
Cartridge
Extraction
Procedure
24
3
1.
Target
Compound
Selection
and
Instrument
Optimization
1.1
Instrument
Configuration.
Before
beginning
the
analysis
of
the
potential
target
compounds
for
the
Unregulated
Contaminant
Monitoring
Rule
(
UCMR)
Gas
Chromatography/
Mass
Spectrometry
(
GC/
MS)
method,
the
instrumentation
needed
to
be
configured
properly.
The
method
was
to
be
developed
on
an
Hewlett
Packard
Model
6890
gas
chromatography
(
GC)
equipped
with
a
Model
5973
mass
selective
detector
(
MSD).
A
standard
split/
splitless
injection
port
was
configured
with
a
2­
mm
i.
d.
quartz,
straight
deactivated
liner.
A
DB­
5MS
capillary
column
(
30m
x
0.25
mm
i.
d.
x
0.25
um
film)
was
selected
for
analyte
separation.
The
system
was
configured
to
operate
in
the
splitless
mode,
with
a
split
delay
of
1
min.

1.2
Target
Compound
List
Evaluation.
The
initial
target
compound
list,
which
is
shown
in
Table
1
below,
was
proposed
by
EPA
for
the
survey.
Initial
analyses
performed
on
the
instrument
were
designed
to
evaluate
signal­
to­
noise
information
and
to
compile
the
mass
spectral
library
for
each
target
analyte.
At
this
time,
elution
order
of
the
targets
was
determined
and
a
retention
time
database
was
compiled
in
the
GC
method.
During
this
phase
of
the
analyses
it
was
determined
that
certain
compounds
would
have
to
be
excluded
from
the
method
due
to
solubility
difficulties
of
some
of
the
neat
materials
and
commercial
availability
of
pure
reference
materials.
Additionally,
the
presence
of
multiple
isomers
in
some
of
the
available
standard
mixtures
added
a
significant
layer
of
complexity
to
quantitation
and
chromatographic
separation,
and
as
a
result,
several
targets
were
eliminated
from
the
method
analyte
list.
These
compounds
were;
allethrin,
cyfluthrin,
cypermethrin,
amitrole,
sulfentrazone,
and
halofenozide.
BDE­
209,
having
a
molecular
weight
over
900
was
not
compatible
with
the
rest
of
the
target
analytes.
To
effectively
analyze
this
compound,
a
column
with
a
higher
temperature
range
would
have
been
required.
For
example
the
DB­
5MS
has
an
recommended
upper
limit
of
325
oC.
BDE­
209
did
not
elute
even
with
a
15­
minute
hold
at
320
oC.
Therefore,
BDE­
209
was
eliminated
from
the
list.
Table
1
depicts
the
results
of
the
single
component
analyses,
chromatographic
observances,
and
issues
encountered
with
the
reference
materials.

Table
1.
Initial
single
component
analysis.

Class
Mult.
Peaks
Good
Peak
2
Peaks
Status
&
Comments
Bromacil
uracil
x
No
Problems
Chlorpyrifos
organophosphate
x
No
Problems
Deltamethrin
pyrethroid
x
No
Problems
Desethyl
Atrazine
triazine
x
No
Problems
Dicofol
chlor.
hydrocarb
x
No
Problems
Esbiol
pyrethroid
x
No
Problems
Hexazinone
triazine
x
No
Problems
Kepone
chlor.
ketone
x
No
Problems
Fenamiphos
organophosphate
x
No
Problems
Malathion
organophosphate
x
No
Problems
Norflurazon
pyridazinone
x
No
Problems
Parathion
organophosphate
x
No
Problems
Prometryn
triazine
x
No
Problems
Terbufos
sulfone
organophosphate
x
No
Problems
Thiobencarb
chloroben
x
No
Problems
4
Class
Mult.
Peaks
Good
Peak
2
Peaks
Status
&
Comments
thiocarbamate
Dimethoate
organophosphate
x
No
Problems
Vinclozolin
dicarboximide
x
No
Problems
BDE­
47
BDE
flame
retardant
x
No
Problems
BDE­
99
BDE
flame
retardant
x
No
Problems
BDE­
100
BDE
flame
retardant
x
No
Problems
BDE­
153
BDE
flame
retardant
x
No
Problems
Mirex
chlorinated
hydrocarbon
x
No
Problems
Nitrophen
nitrophenyl
ether
x
No
Problems
TBBPA
brominated
flame
retard.
x
No
Problems
Allethrin
pyrethroid
x
Std
is
isomer
mixture
(
4­
5
isomers)

Cyfluthrin
pyrethroid
x
Std
is
isomer
mixture
(
4­
5
isomers)

Cypermethrin
pyrethroid
x
Std
is
isomer
mixture
(
4­
5
isomers)

Permethrin
pyrethroid
x
Std
is
isomer
mixture(
1
isomer),
Cis
&
Trans
Phenothrin
pyrethroid
x
Std
is
isomer
mixture(
1
isomer),
Cis
&
Trans
Fenvalerate
pyrethroid
x
Std
is
isomer
mixture(
1
isomer)

Resmethrin
pyrethroid
x
Std
is
isomer
mixture(
1
isomer)

Tetramethrin
pyrethroid
x
Std
is
isomer
mixture(
1
isomer)

Desisopropyl
Atrazine
triazine
Not
Soluble
in
Ethyl
Acetate,
aquired
std
from
Accustd
Propazine
(
Prozinex)
triazine
Not
Soluble
in
Ethyl
Acetate,
aquired
std
from
Accustd
BDE­
209
BDE
flame
retardant
Requires
high
temperature
column
 
did
not
elute
at
upper
practical
limit
for
column
Amitrole
aminotriazole
Aquired
std
from
Accustd
Bifenthrin
pyrethroid
x
No
Problems
Ethofenprox
pyrethroid
x
No
Problems
Sulfentrazone
aryl
triazolinone
x
Trouble
aquiring
standard
above
10ug/
ml
Thiazopyr
pyridinecarboxylate
x
No
Problems
Tralomethrin
pyrethroid
x
Coelutes
with
Deltamethrin,
also
has
same
spectra
Esfenvalerate
pyrethroid
x
Standard
contains
fenvalerate
also
Hexabromobip
henyl
PBB
flame
retard.
Standard
available
as
isomer
mix
Oxychlordane
chlor.
Phenyl
x
No
Problems
HBCD
brominated
flame
retard.
x
Standard
available
as
isomer
mix
Halofenozide
hydrazide
Having
trouble
aquiring
comercially
2.
Extraction
Optimization
2.1
Solid
Phase
Extraction
(
SPE)
Evaluation.
After
finalizing
the
preliminary
target
analyte
list,
an
evaluation
of
several
solid
phase
extraction
materials
was
investigated.
Initially
6
different
solid
phase
materials
were
evaluated,
three
of
which
were
disks
and
three
of
which
were
cartridges.
In
each
case,
five
replicate
reagent
waters
were
fortified
at
5
ug/
L
with
the
target
compounds
and
extracted
using
each
SPE.
Each
SPE
sorbent
was
conditioned
according
the
manufacturers'
suggestions.
The
extractions
were
performed
using
similar
elution
solvents
and
volumes,
and
treated
identically
following
elution
off
of
the
solid
phase.
5
The
solid
phase
that
yielded
the
best
results
was
the
47­
mm
polystyrenedivinylbenzene
(
SDVB)
disk.
All
the
compounds
exhibited
good
extraction
efficiencies
with
the
exception
of
the
atrazine
degradation
products
and
fenamiphos
(
Table
2.).
Fenamiphos
is
known
to
be
problematic
for
standard
stability,
injection
port
stability,
and
extraction
difficulties.
It
is
contained
in
other
EPA
methods
and
therefore
it
is
not
a
primary
concern.
The
atrazine
degradates
(
DIA
&
DEA)
results
were
best
when
extracted
using
the
Oasis­
HLB
cartridge,
but
this
phase
exhibited
poor
recoveries
for
many
other
targets.
It
was
decided
to
remove
the
atrazine
degradates
from
the
analyte
list
and
SDVB
was
selected
as
the
solid
phase.

Table
2.
The
evaluation
of
solid
phase
extraction
sorbents.

SDB
C­
18
SDB­
RPS
Oasis­
HLB
Varianabselut
Phenomenexstrata
Extraction
Media
47
mm
disk
47mm
disk
47mm
disk
cartridge
cartridge
cartridge
Cartridge
6cc/
200mg
10cc/
60mg
6cc/
200mg
Sample
Volume
1000
mL
1000
mL
1000
mL
1000
mL
300
mL
1000
mL
Elution
solvent
volume
15
mL
15
mL
13
mL
13
mL
13
mL
13
mL
Elution
Solvents
Ethyl
Acetate/
CH2Cl2
Ethyl
Acetate/
CH2Cl2
Ethyl
Acetate
Ethyl
Acetate/
CH2Cl2
Ethyl
Acetate/
CH2Cl2
Ethyl
Acetate/
CH2Cl2
Ave
%
Rec
Ave
%
Rec
Ave
%
Rec
Ave
%
Rec
Ave
%
Rec
Ave
%
Rec
SURR
1,3­
dimethyl­
2­
nitrobenzene
106%
76%
108%
112%
100%
128%

Triphenylphosphate
108%
94%
102%
115%
111%
113%
Perylene­
d12
132%
113%
40%
16%
11%
7%
TARGETS
Desisopropyl­
Atrazine
16%
12%
45%
87%
28%
65%
Desethyl­
Atrazine
50%
20%
50%
90%
70%
90%
Dimethoate
71%
19%
77%
96%
70%
97%
Atrazine
57%
100%
99%
99%
Propazine
94%
83%
67%
103%
103%
107%
Vinclozolin
99%
88%
102%
109%
108%
109%
Prometryn
91%
91%
17%
103%
100%
103%
Bromacil
100%
82%
89%
106%
95%
103%
Thiazopyr
99%
86%
93%
107%
105%
91%
Malathion
98%
84%
98%
108%
108%
107%
Chlorpyrifos
94%
83%
95%
101%
89%
99%
Thiobencarb
94%
83%
91%
100%
99%
99%
Parathion
94%
79%
93%
105%
104%
106%
Terbufos­
sulfone
98%
84%
96%
107%
107%
105%
Oxychlordane
97%
84%
93%
95%
57%
92%
Esbiol
96%
86%
93%
97%
92%
86%
Fenamiphos
38%
23%
10%
79%
45%
65%
TBBPA
99%
96%
78%
86%
85%
94%
Ntirophen
102%
88%
95%
112%
99%
106%
Kepone
105%
99%
96%
92%
107%
105%
Norflurazon
102%
95%
98%
113%
107%
114%
6
SDB
C­
18
SDB­
RPS
Oasis­
HLB
Varianabselut
Phenomenexstrata
Hexazinone
95%
95%
85%
98%
94%
102%
Resmethrin
isomer
72%
63%
65%
57%
18%
53%
Resmethrin
71%
65%
63%
51%
15%
45%
Tetramethrin
isomer
105%
90%
92%
95%
86%
89%
Bifenthrin
94%
79%
46%
14%
9%
9%
Tetramethrin
96%
81%
83%
85%
77%
75%
Dicofol
97%
79%
80%
85%
55%
75%
BDE­
47
84%
76%
88%
96%
66%
12%
Phenothrin
isomer
111%
103%
67%
55%
18%
38%
Phenothrin
95%
83%
58%
37%
16%
28%
Mirex
91%
76%
42%
10%
4%
7%
Permethrin
isomer
101%
89%
53%
29%
14%
20%
BDE­
100
90%
88%
74%
75%
65%
10%
Permethrin
105%
97%
57%
34%
14%
21%
BDE­
99
90%
88%
74%
75%
64%
10%
Ethofenprox
134%
125%
91%
99%
63%
49%
HBB
105%
93%
74%
72%
63%
5%
Fenvalerate
104%
92%
44%
22%
14%
14%
Esfenvalerate
100%
84%
41%
19%
11%
12%
BDE­
153
102%
94%
73%
71%
62%
6%
Tralo/
Deltamethrin
111%
94%
39%
17%
10%
10%
HBCD
128%
106%
84%
78%
55%
43%

 
Shaded
areas
indicate
the
highest
recovery
for
the
analyte
relative
to
the
data
set.

3.
Preservation
3.1
pH
and
Antimicrobial
Studies.
Studies
were
conducted
to
investigate
the
effects
of
pH
on
the
target
analyte
list.
Experiments
were
designed
to
evaluate
the
effects
of
acidic,
basic,
and
neutral
buffering
reagents.
Four
pH
levels
were
evaluated,
3.8,
5.0,
7.0,
and
9.0
by
fortifying
buffered
reagent
water
samples
with
target
analytes
at
a
concentration
of
5
ug/
L.
A
fortified
(
unbuffered)
reagent
water
was
prepared
and
analyzed
as
a
control.
The
samples
were
held
for
7
days
at
room
temperature.
The
data
indicated
that
our
analytes
were
most
stable
at
a
low
pH
(
Table
3).
The
results
also
revealed
that
several
of
the
pyrethroid
pesticides
degraded
almost
completely
after
7
days
regardless
of
the
pH.
Additional
literature
research
of
pyrethroid
molecules
researched
revealed
that
several
of
the
pesticides
are
known
to
degrade
when
exposed
to
ultraviolet
light.
To
test
this
theory,
an
experiment
was
designed
to
compare
samples
that
were
stored
for
7
days
in
amber
bottles
vs.
samples
that
were
stored
for
7
days
in
clear
bottles.
Regent
waters
were
again
buffered
and
fortified
with
targets
at
5
ug/
L.
All
samples
were
kept
at
room
temperature
and
buffered
to
a
pH
of
3.8
with
citric
acid
buffer.
The
results
were
conclusive
and
indicated
that
the
low
recovery
for
the
pyrethroids
was
due
to
photo­
degradation
(
Table
4).
Through
the
use
of
a
buffering
solution
to
obtain
a
pH
of
<
4
and
by
storing
the
samples
in
amber
bottles
we
achieved
acceptable
results
for
the
analyte
list.
Based
on
results
obtained
during
the
development
of
Method
531.2,
a
pH
of
<
4
should
also
provide
adequate
protection
against
microbial
degradation.
This
will
be
investigated
later
in
our
holding
time
study.
7
Table
3.
Sample
storage
stability
as
a
function
of
pH
for
samples
held
seven
days
at
room
temperature.

Control
Day
7
pH
3.8
day
7
pH
5
day
7
pH
7
Day
7
pH
9
day7
Ave
%
Rec
Ave
%
Rec
Ave
%
Rec
Ave
%
Rec
Ave
%
Rec
SURR
1,3­
dimethyl­
2­
nitrobenzene
77%
84%
76%
83%
82%

Triphenylphosphate
82%
97%
85%
79%
63%

Perylene­
d12
63%
77%
67%
61%
64%

TARGETS
Dimethoate
73%
87%
75%
74%
48%

Atrazine
100%
97%
94%
103%
106%

Propazine
97%
90%
96%
103%
105%

Vinclozolin
38%
100%
54%
14%
11%

Prometryn
80%
97%
88%
92%
96%

Bromacil
86%
114%
89%
90%
89%

Thiazopyr
84%
105%
94%
94%
80%

Malathion
84%
97%
86%
67%
7%

Chlorpyrifos
83%
93%
83%
84%
82%

Thiobencarb
87%
99%
89%
90%
91%

Parathion
82%
98%
84%
86%
88%

Terbufos­
sulfone
85%
101%
86%
87%
69%

Oxychlordane
90%
99%
88%
87%
87%

Esbiol
75%
96%
79%
75%
65%

Fenamiphos
49%
69%
78%
101%
135%

TBBPA
79%
112%
74%
73%
59%

Ntirophen
88%
107%
84%
85%
86%

Kepone
91%
98%
61%
52%
20%

Norflurazon
88%
105%
88%
84%
83%

Hexazinone
92%
96%
91%
91%
85%

Resmethrin
isomer
1%
0%
7%
7%
0%

Resmethrin
4%
0%
4%
0%
1%

Tetramethrin
isomer
42%
91%
58%
0%
0%

Bifenthrin
80%
93%
86%
84%
87%

Tetramethrin
31%
85%
53%
7%
8%

Dicofol
78%
118%
94%
24%
0%

BDE­
47
67%
73%
70%
69%
80%

Phenothrin
isomer
0%
17%
6%
2%
0%

Phenothrin
4%
12%
5%
6%
2%

Mirex
83%
90%
86%
85%
88%

Permethrin
isomer
86%
99%
83%
81%
81%

BDE­
100
75%
88%
71%
64%
65%

Permethrin
93%
104%
105%
104%
107%

BDE­
99
75%
88%
71%
64%
65%

Ethofenprox
84%
106%
81%
82%
79%

HBB
72%
84%
59%
55%
56%

Fenvalerate
83%
97%
66%
89%
85%

Esfenvalerate
76%
93%
62%
46%
39%

BDE­
153
74%
87%
56%
52%
52%

Tralo/
Deltamethrin
69%
85%
57%
41%
24%
8
Control
Day
7
pH
3.8
day
7
pH
5
day
7
pH
7
Day
7
pH
9
day7
Ave
%
Rec
Ave
%
Rec
Ave
%
Rec
Ave
%
Rec
Ave
%
Rec
HBCD
65%
88%
41%
38%
32%

Average
Recovery
70%
87%
70%
64%
60%

Ave.
CCC
std
recovery
89%
93%
95%
95%
94%

Table
4.
Investigation
of
pyrethroid
compound
stability
stored
in
amber
or
clear
bottles.

pH
3.8
day
0
Clear
pH
3.8
day
0
Amber
pH
3.8
day7
Clear
pH
3.8
day7
Amber
Ave
%
Rec
Ave
%
Rec
Ave
%
Rec
Ave
%
Rec
SURR
1,3­
dimethyl­
2­
nitrobenzene
73%
84%
84%
83%
Triphenylphosphate
98%
91%
89%
86%
Perylene­
d12
94%
87%
76%
78%
TARGETS
Dimethoate
85%
84%
81%
81%
Atrazine
101%
106%
91%
88%
Propazine
99%
102%
91%
87%
Vinclozolin
96%
88%
92%
90%
Prometryn
96%
96%
96%
91%
Bromacil
110%
104%
105%
102%
Thiazopyr
105%
99%
99%
97%
Malathion
99%
96%
94%
91%
Chlorpyrifos
97%
93%
91%
87%
Thiobencarb
98%
94%
95%
90%
Parathion
98%
94%
94%
92%
Terbufos­
sulfone
101%
96%
95%
92%
Oxychlordane
104%
102%
98%
93%
Esbiol
102%
99%
95%
96%
Fenamiphos
73%
87%
68%
60%
TBBPA
140%
127%
113%
105%
Ntirophen
108%
102%
102%
99%
Kepone
89%
86%
78%
70%
Norflurazon
105%
98%
99%
93%
Hexazinone
91%
93%
97%
88%
Resmethrin
isomer
76%
106%
0%
97%
Resmethrin
79%
110%
4%
99%
Tetramethrin
isomer
104%
103%
98%
103%
Bifenthrin
97%
96%
95%
94%
Tetramethrin
97%
97%
90%
93%
Dicofol
123%
118%
114%
114%
BDE­
47
104%
77%
89%
76%
Phenothrin
isomer
107%
100%
23%
104%
Phenothrin
101%
100%
26%
97%
Mirex
92%
92%
89%
85%
Permethrin
isomer
104%
100%
101%
97%
9
pH
3.8
day
0
Clear
pH
3.8
day
0
Amber
pH
3.8
day7
Clear
pH
3.8
day7
Amber
Ave
%
Rec
Ave
%
Rec
Ave
%
Rec
Ave
%
Rec
BDE­
100
118%
94%
95%
83%
Permethrin
103%
99%
108%
97%
BDE­
99
118%
94%
95%
83%
Ethofenprox
124%
117%
119%
118%
HBB
116%
90%
90%
78%
Fenvalerate
109%
103%
99%
96%
Esfenvalerate
99%
93%
91%
88%
BDE­
153
115%
88%
95%
80%
Tralo/
Deltamethrin
96%
85%
89%
82%
HBCD
102%
82%
77%
78%
Average
Recovery
102%
97%
87%
91%

3.1
Dechlorination.
Many
of
the
finished
drinking
waters
contain
residual
free
available
chlorine
(
FAC),
which
can
degrade
target
analytes.
This
was
investigated
next.
The
first
experiments
conducted
evaluated
sodium
sulfite,
sodium
thiosulfate,
and
glycine
as
potential
reagents
in
the
absence
of
free
available
chlorine.
These
data
indicate
that
sodium
thiosulfate
produced
the
best
results
for
the
target
analyte
list
(
Table
5).
It
was
also
noted
that
both
glycine
and
sodium
sulfite
caused
analyte
degradation,
most
notably
to
the
pyrethroids.
One
side
effect
of
sodium
thiosulfate
being
used
as
the
dechlorinating
reagent
was
it
produced
extraneous
elemental
sulfur
peaks
in
the
GC
chromatogram.
These
peaks
had
the
potential
to
cause
interferences
with
quantitation
and
identification.
Because
of
this,
the
next
study
evaluated
a
fourth
potential
microbial
inhibitor,
ascorbic
acid.
This
next
study
also
examined
the
effect
of
free
chlorine
on
the
analytes
in
the
presence
and
absence
of
a
dechlorinating
reagent.
Results
were
compared
from
samples
that
had
100
mg
of
sodium
thiosulfate
+
6mg/
L
of
free
chlorine,
and
samples
that
contained
100
mg/
L
of
ascorbic
acid
+
6mg/
L
of
free
chlorine
to
samples
that
just
contained
6
mg/
L
of
free
chlorine
(
Table
6).
Again
the
samples
were
held
at
room
temperature
for
a
period
of
7
days
before
extracting.
It
was
found
that
the
presence
of
free
chlorine
in
the
samples
began
to
degrade
some
of
the
analytes.
It
was
also
found
that
absorbic
acid
was
as
equally
effective
as
sodium
thiosulfate
as
a
dechlorinating
reagent
but
did
not
produce
the
sulfur
peaks
in
the
GC
chromatogram.
The
use
of
ascorbic
acid
led
to
an
improvement
in
the
ruggedness
of
the
method.
After
several
samples
were
analyzed
that
were
dechlorinated
with
sodium
thiosulfate,
degradation
in
the
injection
port
was
observed
sooner
than
expected
(
after
40­
50
samples).
We
noticed
that
the
gold
inlet
seal
was
turning
black
from
the
excess
sulfur
burning
off
in
the
injection
port.
Ascorbic
acid
did
not
produce
this
inlet
effect
and
injection
port
maintenance
was
not
necessary
as
often
(
150­
250
samples).
10
Table
5.
The
evaluations
of
dechlorinating
reagents
in
the
absence
of
free
available
chlorine.
All
targets
were
fortified
into
reagent
water
at
5
ug/
L.

Control
50
mg
sodium
sulfite
100
mg
sodium
thiosulfate
250
mg
glycine
Ave
%
Rec
Ave
%
Rec
Ave
%
Rec
Ave
%
Rec
SURR
1,3­
dimethyl­
2­
nitrobenzene
83%
109%
98%
83%
Triphenylphosphate
102%
112%
105%
97%
Perylene­
d12
86%
4%
91%
78%
TARGETS
Dimethoate
73%
87%
82%
10%
Atrazine
87%
94%
91%
87%
Propazine
86%
90%
92%
84%
Vinclozolin
114%
121%
122%
122%
Prometryn
95%
25%
36%
57%
Bromacil
106%
123%
109%
107%
Thiazopyr
101%
96%
105%
90%
Malathion
94%
102%
99%
31%
Chlorpyrifos
95%
99%
100%
48%
Thiobencarb
95%
104%
100%
86%
Parathion
107%
129%
112%
44%
Terbufos­
sulfone
98%
105%
101%
27%
Oxychlordane
98%
100%
104%
92%
Esbiol
99%
11%
103%
9%
Fenamiphos
22%
21%
99%
0%
TBBPA
97%
103%
105%
88%
Ntirophen
109%
132%
119%
116%
Kepone
99%
104%
105%
97%
Norflurazon
101%
110%
107%
98%
Hexazinone
95%
76%
98%
93%
Resmethrin
isomer
92%
10%
85%
6%
Resmethrin
95%
11%
89%
8%
Tetramethrin
isomer
95%
15%
92%
17%
Bifenthrin
90%
92%
91%
90%
Tetramethrin
90%
11%
91%
8%
BDE­
47
82%
96%
89%
83%
Phenothrin
isomer
93%
47%
88%
0%
Phenothrin
93%
53%
93%
13%
Mirex
91%
90%
92%
92%
Permethrin
isomer
95%
99%
95%
95%
BDE­
100
87%
93%
94%
86%
Permethrin
90%
98%
88%
91%
BDE­
99
88%
100%
95%
86%
Ethofenprox
94%
116%
101%
90%
HBB
86%
99%
99%
83%
Fenvalerate
99%
102%
103%
90%
Esfenvalerate
96%
103%
103%
87%
BDE­
153
86%
100%
101%
81%
11
Control
50
mg
sodium
sulfite
100
mg
sodium
thiosulfate
250
mg
glycine
Ave
%
Rec
Ave
%
Rec
Ave
%
Rec
Ave
%
Rec
Tralo/
Deltamethrin
94%
111%
105%
84%
HBCD
90%
97%
100%
69%
Average
Recovery
92%
84%
97%
66%

Table
6.
Comparison
of
ascorbic
acid
and
sodium
thiosulfate
as
dechlorinating
reagents
for
reagent
water
samples
fortified
at
5
ug/
L
and
then
held
at
room
temperature
for
7
days.

Control
FAC
@
6
mg/
L
100
mg/
L
Ascorbic
acid
+
6
mg/
L
FAC
100
mg/
L
Sodium
thiosulfate
+
6mg/
L
FAC
SURR
1,3­
dimethyl­
2­
nitrobenzene
93%
81%
83%
84%

Triphenylphosphate
94%
87%
94%
82%

Perylene­
d12
97%
89%
97%
90%
TARGETS
Dimethoate
74%
39%
81%
58%

Atrazine
76%
67%
81%
53%

Propazine
76%
62%
82%
55%

Vinclozolin
99%
93%
101%
102%

Prometryn
93%
57%
105%
64%
Bromacil
106%
99%
112%
110%

Thiazopyr
103%
87%
102%
97%

Malathion
99%
60%
102%
52%

Chlorpyrifos
87%
54%
90%
84%

Thiobencarb
96%
67%
97%
94%

Parathion
119%
59%
123%
136%
Terbufos­
sulfone
97%
59%
98%
78%

Oxychlordane
91%
87%
95%
88%

Esbiol
105%
49%
103%
91%

Fenamiphos
48%
0%
117%
49%

TBBPA
99%
95%
107%
99%

Ntirophen
108%
97%
103%
76%
Kepone
91%
98%
111%
85%

Norflurazon
104%
97%
108%
95%

Hexazinone
88%
78%
93%
66%

Resmethrin
isomer
91%
63%
96%
60%

Resmethrin
80%
64%
88%
56%

Tetramethrin
isomer
128%
57%
123%
62%
Bifenthrin
101%
90%
109%
58%

Tetramethrin
105%
47%
105%
61%

Dicofol
118%
107%
126%
92%
12
Control
FAC
@
6
mg/
L
100
mg/
L
Ascorbic
acid
+
6
mg/
L
FAC
100
mg/
L
Sodium
thiosulfate
+
6mg/
L
FAC
BDE­
47
78%
78%
105%
98%

Phenothrin
isomer
114%
80%
115%
105%

Phenothrin
100%
80%
108%
93%

Mirex
100%
91%
109%
91%
Permethrin
isomer
98%
85%
106%
83%

BDE­
100
83%
81%
105%
108%

Permethrin
115%
105%
126%
98%

BDE­
99
82%
79%
105%
106%

Ethofenprox
113%
98%
124%
113%

HBB
81%
80%
102%
98%
Fenvalerate
120%
100%
137%
63%

Esfenvalerate
99%
92%
111%
52%

BDE­
153
72%
71%
97%
84%

Tralo/
Deltamethrin
121%
109%
125%
70%

HBCD
107%
117%
124%
121%
Average
Recovery
97%
78%
106%
83%

3.3
Degradation/
Hydrolysis
Studies.
The
presence
of
transition
metal
ions
can
increase
the
rate
of
hydrolysis
for
some
organic
molecules.
The
presence
of
Cu++
ions
in
drinking
water
can
be
a
common
occurrence
due
to
the
use
of
copper
tubing
for
drinking
water
distribution
lines.
The
addition
of
ethylenediametetraacetic
acid
(
EDTA)
to
the
samples
can
eliminate
the
copper
ions
from
the
matrix
and
thereby
eliminate
the
potential
increased
rate
of
hydrolysis
of
the
organic
analytes.
To
determine
the
effects
of
Cu++
and
EDTA
on
the
target
analytes,
the
following
experiments
were
performed.
Four
sets
of
reagent
water
samples
were
fortified
at
5
ug/
L,
buffered
to
a
pH
of
3.8,
and
held
at
room
temperature
for
7
days
in
amber
bottles.
Each
sample
also
was
fortified
with
6
mg/
L
of
free
chlorine
that
had
been
dechlorinated
with
100
mg/
L
of
ascorbic
acid
prior
to
fortification
with
the
target
compounds.
In
one
set
of
samples
1.3
mg/
L
of
Cu++
was
added
as
copper
sulfate
(
1.3
mg/
L
of
Cu++
was
used
as
a
"
high"
level
because
it
is
above
the
regulatory
limit
of
Cu++
allowed
in
safe
drinking
water).
The
second
set
of
samples
contained
0.30
mg/
L
of
Cu++.
To
the
third
set
we
added
1.3
mg/
L
of
Cu++
and
350
mg/
L
of
EDTA.
In
addition
to
these
sets
we
ran
a
control
for
the
experiment.
Reagents
added
to
each
sample
can
be
found
at
the
bottom
of
the
data
table
(
Table
7.)
The
purpose
of
fortifying
two
extraction
sets
at
different
levels
of
Cu++
was
to
determine
the
effects
of
a
high
content
and
a
low
content
of
Cu++
on
the
degradation
of
the
analytes.
As
can
be
seen
from
Table
7,
EDTA
is
required
if
transition
metals
like
copper
are
present
in
drinking
water.
13
Table
7.
The
evaluation
of
the
need
for
protection
from
transition
metal­
catalyzed
hydrolysis
using
EDTA.

Control
EDTA
&
Cu++
(
1.3mg/
L)
Cu++
only
(
0.3mg/
L)
Cu++
only
(
1.3mg/
L)

Ave
%
Rec
Ave
%
Rec
Ave
%
Rec
Ave
%
Rec
SURR
1,3­
dimethyl­
2­
nitrobenzene
85%
93%
62%
65%
Triphenylphosphate
95%
108%
68%
61%
Perylene­
d12
96%
90%
81%
80%
TARGETS
Dimethoate
67%
76%
49%
43%
Atrazine
79%
87%
68%
65%
Propazine
73%
80%
62%
61%
Vinclozolin
88%
104%
63%
64%
Prometryn
92%
98%
80%
75%
Bromacil
98%
103%
80%
68%
Thiazopyr
89%
97%
43%
59%
Malathion
84%
92%
58%
52%
Chlorpyrifos
81%
93%
51%
55%
Thiobencarb
85%
99%
64%
59%
Parathion
92%
101%
60%
54%
Terbufos­
sulfone
89%
95%
62%
55%
Oxychlordane
97%
98%
91%
81%
Esbiol
86%
98%
49%
45%
Fenamiphos
102%
92%
76%
66%
TBBPA
111%
108%
103%
79%
Ntirophen
100%
104%
74%
72%
Kepone
94%
98%
82%
82%
Norflurazon
96%
106%
72%
69%
Hexazinone
97%
105%
80%
70%
Resmethrin
isomer
103%
108%
68%
56%
Resmethrin
105%
105%
76%
70%
Tetramethrin
isomer
93%
93%
13%
20%
Bifenthrin
89%
85%
84%
80%
Tetramethrin
81%
84%
11%
17%
BDE­
47
84%
88%
81%
83%
Phenothrin
isomer
103%
97%
93%
68%
Phenothrin
89%
89%
77%
70%
Mirex
84%
85%
80%
83%
Permethrin
isomer
94%
93%
87%
79%
BDE­
100
86%
86%
81%
82%
14
Control
EDTA
&
Cu++
(
1.3mg/
L)
Cu++
only
(
0.3mg/
L)
Cu++
only
(
1.3mg/
L)

Ave
%
Rec
Ave
%
Rec
Ave
%
Rec
Ave
%
Rec
Permethrin
93%
91%
85%
74%
BDE­
99
86%
94%
84%
84%
Ethofenprox
101%
99%
95%
82%
HBB
92%
92%
90%
89%
Fenvalerate
104%
91%
99%
82%
Esfenvalerate
97%
90%
94%
80%
BDE­
153
97%
89%
90%
85%
Tralo/
Deltamethrin
99%
93%
88%
83%
HBCD
118%
93%
118%
not
spiked
Average
Recovery
92%
95%
74%
68%

Sample
Conditions
6mg/
L
free
Chlorine
6mg/
L
free
Chlorine
6mg/
L
free
Chlorine
6mg/
L
free
Chlorine
Buffered
to
pH
3.8
Buffered
to
pH
3.8
Buffered
to
pH
3.8
Buffered
to
pH
3.8
100mg
Absorbic
Acid
100mg
Absorbic
Acid
100mg
Absorbic
Acid
100mg
Absorbic
Acid
7­
day
hold
@
room
temp.
7­
day
hold
@
room
temp.
7­
day
hold
@
room
temp.
7­
day
hold
@
room
temp.

1.3mg/
L
Cu++
0.3mg/
L
Cu++
1.3mg/
L
Cu++
350mg
EDTA
4.
Ruggedness.
Every
effort
was
made
to
produce
an
analytical
method
that
was
rugged.
Many
issues
related
to
the
robustness
of
the
method
were
solved
through
the
selection
and
optimization
of
the
correct
preservation
materials.
A
few
issues,
however,
required
additional
troubleshooting.

4.1
Syringe
Issues.
Problems
were
encountered
with
the
autosampler
syringe
erring
repeatedly
due
to
a
"
stickiness"
that
was
building
up
inside
the
syringe
barrel.
Ultimately
this
issue
was
resolved
by
adding
additional
autosampler
rinses
including
automated
methanol
rinses
of
the
syringe
in
between
injections.
We
found
that
rinsing
the
syringe
with
methylene
chloride
and
methanol
in
between
sample
injections
corrected
the
problem.

4.2
BDE
Precision.
While
acceptable
results
for
accuracy
and
extraction
efficiency
of
the
BDE
compounds
was
achieved,
less
than
desirable
precision
was
observed
on
several
occasions.
RSDs
were
approximately
10%
and
occasionally
reached
15­
20
%
RSD
(
n=
5)
during
the
precision
and
accuracy
studies.
Through
further
research
it
was
discovered
that
the
imprecision
was
likely
due
to
either
a
mass
defect
issue
in
the
MS,
or
too
low
of
an
injection
port
temperature.
The
isotopic
mass
for
the
tetra­,
penta­,
hexa­
and
decabrominated
PBBEs
are
481.7,
559.6,
637.5,
and
949.2,
respectively.
If
one
were
to
use
these
ions
for
quantitation
(
or
their
associated
M+
2,
M+
4
or
even
the
M+
6
ions)
mass
15
defect
could
result
in
precision
issues
if
nominal
masses
(
whole
numbers)
were
used
for
generating
extracted
ion
profiles
for
quantitation.
In
practice,
Method
527
uses
masses
assocaited
with
the
loss
of
two
bromines
for
all
but
BDE­
153.
Masses
in
the
method
for
the
BDEs
have
been
expressed
to
four
significant
figures
and
a
note
added
regarding
this
potential
issue.

An
injection
port
temperature
study
was
performed
to
determine
the
optimal
temperature
for
precision
(
Table
8).
We
found
that
an
increase
in
temperature
to
250
°
C
improved
precision
and
response
of
the
BDEs
but
did
not
negatively
effect
the
other
analytes
on
the
target
compound
list
for
the
Agilent
instrument.
Higher
injection
port
temperatures
did
case
some
loss
in
sensitivity
and
precision
for
the
early
eluting
compounds.
These
same
studies
were
repeated
on
the
Saturn
2000.
It
appears
that
differences
in
the
injection
port
geometry,
heating
profiles,
and
inertness
results
in
different
ideal
temperatures
for
the
port
on
each
instrument.
A
lower
injection
port
temperature
on
the
Varian
instrument
has
a
much
greater
effect
on
results
than
on
the
HP
injection
port.
(
Table
9).
A
temperature
of
250
oC
was
chosen
as
a
middle
ground
for
both
systems.

Table
8.
Injection
port
temperature
optimization
using
the
HP
GC/
MS.

HP
6890/
5972
Port
Temp.
BDE­
47
BDE­
100
BDE­
99
Hexabromobiphenyl
BDE­
153
210
Ave
Area
2944333
1432068
1327087
552816
236958
%
RSD
5.6
9.0
10.4
10.7
18.4
230
Ave
Area
3421740
1798830
1804146
790645
362769
%
RSD
2.8
4.3
3.4
6.5
4.2
250
Ave
Area
3335905
1847741
1934847
910335
429059
%
RSD
4.2
4.7
5.9
7.1
10.1
270
Ave
Area
3406209
1884238
2054908
970498
490742
%
RSD
2.5
4.8
5.2
4.2
5.1
290
Ave
Area
3204373
1903584
2009525
957052
469724
%
RSD
2.6
1.3
2.7
2.9
7.8
Table
9.
Injection
port
temperature
optimization
using
the
Saturn
2000.

Var
3800/
2000
Port
Temp.
BDE­
47
BDE­
100
BDE­
99
Hexabromobiphenyl
BDE­
153
210
Ave
Area
1257237
410270
463955
893328
438796
%
RSD
10.5
16.1
21.2
31.1
31.8
230
Ave
Area
1223050
400013
429228
889483
426906
%
RSD
5.6
9.9
13.2
22.9
23.0
250
Ave
Area
1300860
434173
484762
1038823
479284
%
RSD
4.9
3.7
5.3
5.3
4.4
270
Ave
Area
989719
337451
362994
783086
363382
%
RSD
12.9
10.7
12.3
13.8
13.4
290
Ave
Area
952840
331067
337484
695529
326140
%
RSD
10.0
8.5
10.1
12.1
9.0
16
5
Second
Source
Standard
Evaluation.

5.1
Individual
Standard
Analyses.
All
analytes
on
the
target
list
were
analyzed
from
reference
materials
procured
from
two
separate
manufacturers.
From
this
we
were
able
to
verify
that
our
mass
spectral
information
and
the
accuracy
of
the
standard
materials
were
valid.
First
the
instrument
was
calibrated
with
standards
obtained
from
Accustandard
as
ampulized
standard
mixtures.
Mass
spectra
were
obtained
for
each
component
and
an
MS
library
and
retention
time
database
was
compiled.
Next,
each
target
analyte
was
analyzed
as
a
single
component
standard
obtained
from
a
second
source.
Most
standards
were
purchased
as
individual
neat
compounds
from
Chem.
Service
or
from
Fluka.
Mass
spectra
from
the
single
component
analysis
were
then
compared
to
the
ampulized
solutions
for
qualitative
integrity,
and
retention
times
were
compared
against
the
calibration.
All
single
component
standards
were
then
quantitated
using
the
calibrations
obtained
from
the
ampulized
solutions.
Accuracy
was
validated
if
the
compound
results
were
within
70­
130%
recovery
(
Table
9)
where
recovery
is
used
to
refer
to
the
calculated
amount
for
the
second
source
standard
using
the
primary
source
calibration
curve.
For
example,
a
second
source
that
is
fortified
at
an
equivalent
extract
concentration
of
10
ug/
L
would
have
an
acceptance
range
from
7.0
to
13
ug/
L.

5.2
Thiazopyr
and
Ethofenprox.
Two
analytes,
when
analyzed
individually,
did
not
match
the
primary
standards
that
had
been
used
during
method
development.
A
third
source
of
acceptable
purity
was
not
commercially.
As
a
result,
no
determination
could
be
made
regarding
which
standard
was
accurate.
Due
to
this
issue,
both
compounds
were
eliminated
from
the
method
analyte
list.

Table
9.
Second
source
standard
evaluation.

Analyte
%
Rec.
(
n=
3)
Dimethoate
94
Atrazine
110
Propazine
103
Vinclozolin
122
Prometryn
121
Bromacil
96
Malathion
127
Thiazopyr
391
Chlorpyrifos
109
Thiobencarb
103
Parathion
107
Terbufos­
Sulfone
106
Oxychlordane
98
Esbiol
103
Fenamiphos
80
Nitrophen
105
Kepone
109
Norflurazon
95
Hexazinone
96
Bifenthrin
108
2,2',
4,4'­
Tetrabromodiphenyl
Ether
(
BDE­
47)
93
Mirex
119
2,2',
4,4',
6­
Pentabromodiphenyl
Ether
(
BDE­
100)
94
17
Analyte
%
Rec.
(
n=
3)
2,2',
4,4',
5­
Pentabromodiphenyl
Ether
(
BDE­
99)
82
Ethofenprox
268
Hexabromobiphenyl
108
Fenvalerate
84
Esfenvalerate
73
2,2',
4,4',
5,5'­
Hexabromodiphenyl
Ether
(
BDE­
153)
80
6
Precision
and
Accuracy.
Precision
and
accuracy
of
the
method
was
determined
in
three
different
matrices
at
two
fortification
levels.
A
concentration
of
1.0
ug/
L
was
chosen
because
it
was
near
the
MRL
and
5.0
ug/
L
was
chosen
because
it
was
equal
to
a
calibration
point
near
the
mid­
to
high­
end
of
the
established
calibration
range.
The
three
matrices
were
reagent
water,
surface
water,
and
ground
water.
All
samples
were
extracted
and
analyzed
according
to
the
conditions
of
the
method
and
were
extracted
by
solid
phase
extraction
using
a
polystyrenedivinylbenzene
extraction
disk.
Results
of
the
precision
and
accuracy
study
can
be
found
in
Tables
10,
11
and
12.
Method
527
met
all
UCMR
requirements
for
precision
and
accuracy.

Table
10.
Precision
and
accuracy
for
method
analytes
fortified
at
1.0
and
5.0
ug/
L
in
reagent
water
extracted
with
SDVB
disks.

Concentration
=
1.0
ug/
L,
(
n=
5)
Concentration
=
5.0
ug/
L,
(
n=
5)

Analyte
%
Recovery
%
RSD
%
Recovery
%
RSD
Dimethoate
114
5.3
86
4.5
Atrazine
109
12.8
86
8.7
Propazine
95
9.1
82
9.1
Vinclozolin
116
5.0
94
6.0
Prometryn
106
4.1
86
5.6
Bromacil
127
5.1
103
5.7
Malathion
107
2.1
86
4.5
Chlorpyrifos
98
3.3
85
5.4
Thiobencarb
102
1.8
87
4.7
Parathion
107
4.1
88
5.1
Terbufos­
Sulfone
111
4.2
93
4.3
Oxychlordane
106
3.9
84
6.0
Esbiol
119
4.1
97
5.2
Fenamiphos
90
12
73
16
Nitrophen
116
3.5
100
4.6
Kepone
103
4.3
83
4.3
Norflurazon
134
1.9
106
5.8
Hexazinone
120
7.0
94
6.8
Bifenthrin
102
1.8
79
4.3
2,2',
4,4'­
Tetrabromodiphenyl
Ether
(
BDE­
47)
93
5.0
81
4.1
Mirex
93
3.4
75
4.7
2,2',
4,4',
6­
Pentabromodiphenyl
Ether
(
BDE­
100)
93
8.1
84
3.9
2,2',
4,4',
5­
Pentabromodiphenyl
Ether
(
BDE­
99)
98
8.6
87
5.4
Hexabromobiphenyl1
102
5.8
82
3.6
Fenvalerate
120
3.6
96
4.4
Esfenvalerate
111
3.4
86
4.8
2,2',
4,4',
5,5'­
Hexabromodiphenyl
Ether
(
BDE­
153)
96
10
85
9.3
18
Concentration
=
1.0
ug/
L,
(
n=
5)
Concentration
=
5.0
ug/
L,
(
n=
5)

Analyte
%
Recovery
%
RSD
%
Recovery
%
RSD
1,3­
Dimethyl­
2­
Nitrobenzene
(
SUR)
93
3.2
88
9.1
Triphenylphosphate
(
SUR)
88
2.0
88
4.8
Perylene­
d12
(
SUR)
84
4.8
79
5.6
1Precision
and
accuracy
determinations
were
obtained
by
using
a
technical
mixture
of
Hexabromobiphenyl
(
Firemaster
BP­
6).
Actual
spiked
concentrations
for
the
isomer
2,2',
4,4',
5,5'­
Hexabromobiphenyl
are
0.74
ug/
L
and
3.70
ug/
L
respectively
Table
11.
Precision
and
accuracy
for
method
analytes
fortified
at
1.0
and
5.0
ug/
L
in
surface
water
extracted
with
SDVB
disks.

Concentration
=
1.0
ug/
L,
n=
5
Concentration
=
5.0
ug/
L,
n=
5
Analyte
Mean
%
Recovery
%
RSD
Mean
%
Recovery
%
RSD
Dimethoate
112
2.7
89
2.6
Atrazine
106
4.5
102
2.9
Propazine
92
6.3
90
4.1
Vinclozolin
115
4.7
103
6.0
Prometryn
101
2.9
91
4.9
Bromacil
119
2.1
104
3.2
Malathion
106
2.5
95
4.0
Chlorpyrifos
95
2.4
91
4.8
Thiobencarb
96
2.7
95
4.5
Parathion
105
2.1
94
4.7
Terbufos­
Sulfone
107
1.6
97
4.6
Oxychlordane
94
4.9
91
3.4
Esbiol
114
2.6
104
4.8
Fenamiphos
132
8.9
70
11
Nitrophen
111
2.7
99
3.7
Kepone
91
5.0
88
4.0
Norflurazon
122
4.8
105
3.2
Hexazinone
120
2.2
101
3.1
Bifenthrin
97
2.4
90
5.1
2,2',
4,4'­
Tetrabromodiphenyl
Ether
(
BDE­
47)
93
6.3
89
2.9
Mirex
80
5.4
87
5.6
2,2',
4,4',
6­
Pentabromodiphenyl
Ether
(
BDE­
100)
96
5.1
91
1.8
2,2',
4,4',
5­
Pentabromodiphenyl
Ether
(
BDE­
99)
108
4.6
93
2.7
Hexabromobiphenyl1
102
9.2
95
2.5
Fenvalerate
124
3.4
105
3.9
Esfenvalerate
113
2.9
97
3.3
2,2',
4,4',
5,5'­
Hexabromodiphenyl
Ether
(
BDE­
153)
108
6.7
91
5.3
1,3­
Dimethyl­
2­
Nitrobenzene
(
SUR)
80
5.2
97
14
Triphenylphosphate
(
SUR)
83
3.6
98
4.2
Perylene­
d12
(
SUR)
83
3.0
93
4.2
1Precision
and
accuracy
determinations
were
obtained
by
using
a
technical
mixture
of
Hexabromobiphenyl
(
Firemaster
BP­
6).
Actual
spiked
concentrations
for
the
isomer
2,2',
4,4',
5,5'­
Hexabromobiphenyl
are
0.74
ug/
L
and
3.70
ug/
L
respectively.
19
Table
12.
Precision
and
accuracy
for
method
analytes
fortified
at
1.0
and
5.0
ug/
L
in
groundwater
extracted
with
SDVB
disks.

Concentration
=
1.0
ug/
L,
n=
5
Concentration
=
5.0
ug/
L,
n=
4
Analyte
Mean
%
Recovery
%
RSD
Mean
%
Recovery
%
RSD
Dimethoate
114
7.1
78
4.3
Atrazine
120
7.2
90
9.6
Propazine
98
9.8
83
9.5
Vinclozolin
116
6.0
96
8.9
Prometryn
102
8.2
85
9.7
Bromacil
128
7.3
93
9.5
Malathion
106
6.1
85
10
Chlorpyrifos
100
3.5
85
10
Thiobencarb
100
6.4
86
9.6
Parathion
103
4.2
87
9.8
Terbufos­
Sulfone
110
8.1
87
11
Oxychlordane
102
12
86
10
Esbiol
109
8.3
95
8.6
Fenamiphos
150
13
85
6.4
Nitrophen
109
6.3
89
8.4
Kepone
84
6.7
75
7.4
Norflurazon
116
6.4
95
7.8
Hexazinone
114
7.7
93
7.5
Bifenthrin
98
6.0
77
11
2,2',
4,4'­
Tetrabromodiphenyl
Ether
(
BDE­
47)
87
6.8
78
5.6
Mirex
90
8.0
73
12
2,2',
4,4',
6­
Pentabromodiphenyl
Ether
(
BDE­
100)
86
5.8
80
3.8
2,2',
4,4',
5­
Pentabromodiphenyl
Ether
(
BDE­
99)
87
8.9
83
4.2
Hexabromobiphenyl1
88
5.1
80
6.1
Fenvalerate
116
8.4
94
7.2
Esfenvalerate
111
8.2
88
9.4
2,2',
4,4',
5,5'­
Hexabromodiphenyl
Ether
(
BDE­
153)
100
9.8
83
4.6
1,3­
Dimethyl­
2­
Nitrobenzene
(
SUR)
95
5.8
86
12
Triphenylphosphate
(
SUR)
90
4.6
86
9.6
Perylene­
d12
(
SUR)
86
3.3
83
8.5
1Precision
and
accuracy
determinations
were
obtained
by
using
a
technical
mixture
of
Hexabromobiphenyl
(
Firemaster
BP­
6).
Actual
spiked
concentrations
for
the
isomer
2,2',
4,4',
5,5'­
Hexabromobiphenyl
are
0.74
ug/
L
and
3.70
ug/
L
respectively.

7.0
LCMRL
and
MDL
Determinations.

7.1
Detection
Limit
(
DL).
DL
values
were
calculated
for
each
target
analyte
from
the
data
used
to
determine
the
Lowest
Concentration
Minimum
Reporting
Limit.
In
each
case,
the
DL
was
determined
by
extracting
and
analyzing
seven
laboratory
fortified
reagent
blanks.
The
DLs
were
calculated
with
the
following
equation:

DL
=
St(
n
­
1,
1
­
alpha
=
0.99)

where
20
t(
n
­
1,
1
­
alpha
=
0.99)
=
Students
t
value
for
the
99%
confidence
level
with
n­
1
degrees
of
freedom,
n
=
number
of
replicates,
and
S
=
standard
deviation
of
replicate
analyses.

Results
for
the
DL
study
can
be
found
in
Table
13.

7.2
Lowest
Concentration
Minimum
Reporting
Limit
(
LCMRL).
The
single­
laboratory
LCMRL
is
the
lowest
true
concentration
for
which
the
future
recovery
is
predicted
to
fall,
with
high
confidence
(
99
percent),
between
50
and
150
percent
recovery.
Calculating
these
values
involved
preparing,
extracting,
and
analyzing
seven
replicates
at
5
concentrations
(
0.1,
0.2,
0.35,
0.5,
and
1.0
ug/
L)
and
then
calculating
the
LCMRL
using
Sigma­
Plot
software.
LCMRLs,
DLs,
and
estimated
signal­
to­
noise
values
at
the
LCMRL
are
summarized
in
Table
13.

Table
13.
LCMRL
and
DL
values
determined
for
method
analytes.

Analyte
Spiking
Conc.
(
ug/
L)
1
DL
(
ug/
L)
LCMRL
(
ug/
L)
S/
N
@
LCMRL
Dimethoate
0.10
0.025
0.36
7.2
Atrazine
0.10
0.036
0.16
12
Propazine
0.10
0.039
0.18
15
Vinclozolin
0.20
0.084
0.29
15
Prometryn
0.10
0.028
0.20
17
Bromacil
0.20
0.093
0.45
8.6
Malathion
0.20
0.057
0.51
12
Chlorpyrifos
0.10
0.026
0.12
11
Thiobencarb
0.20
0.038
0.13
9.9
Parathion
0.20
0.062
0.29
5.4
Terbufos­
Sulfone
0.10
0.041
0.27
18
Oxychlordane
0.35
0.110
0.27
8.8
Esbiol
0.10
0.041
0.31
15
Fenamiphos
0.50
0.110
1.10
14
Nitrophen
0.20
0.071
0.51
13
Kepone
0.20
0.076
0.35
5.1
Norflurazon
0.20
0.076
0.53
14
Hexazinone
0.10
0.046
0.41
22
Bifenthrin
0.20
0.040
0.21
21
2,2',
4,4'­
Tetrabromodiphenyl
Ether
(
BDE­
47)
0.10
0.028
0.18
6.3
Mirex
0.10
0.022
0.31
13
2,2',
4,4',
6­
Pentabromodiphenyl
Ether
(
BDE­
100)
0.35
0.051
0.29
5.8
2,2',
4,4',
5­
Pentabromodiphenyl
Ether
(
BDE­
99)
0.20
0.097
0.39
6.5
Hexabromobiphenyl
0.37
0.110
0.44
10
Fenvalerate
0.35
0.079
0.67
16
Esfenvalerate
0.20
0.062
0.48
12
2,2',
4,4',
5,5'­
Hexabromodiphenyl
Ether
(
BDE­
153)
0.20
0.140
0.40
11
1Spiking
concentration
used
to
determine
MDL
8.0
Storage
Stability.
The
method
uses
citric
acid
(
9.4
g/
L)
to
acidify
samples
to
approximately
pH
3.8,
uses
ascorbic
acid
(
100
mg/
L)
to
dechlorinate
the
samples,
and
uses
EDTA
(
350
mg/
L)
to
remove
transition
metals
like
copper
that
otherwise
could
21
catalyze
hydrolysis.
The
effectiveness
of
this
preservation
scheme
was
tested
through
a
storage
stability
study
by
extracting
and
analyzing
fortified
surface
waters
preserved
and
held
according
to
the
method
requirements,
and
by
conducting
pour
plate
studies
to
enumerate
viable
microorganisms.
These
experiments,
which
were
conducted
in
triplicate
at
7­
day
increments
over
a
28­
day
period,
evaluated
recovery
of
the
target
analytes
in
the
presence
and
absence
of
the
antimicrobial
agent
(
citric
acid)
and
in
the
presence
and
absence
of
a
microbial
spike
(
4
mL
of
Ohio
River
water
added
to
each
1­
L
sample).
This
experimental
design
is
summarized
in
Table
14.
All
experiments
included
a
method
blank
each
day
for
each
matrix.
All
samples
were
stored
for
2
days
at
10
°
C
then
at
6
°
C
for
the
remainder
of
the
study.
Target
compound
recovery
data
are
presented
in
Table
15.
The
results
of
the
pour
plate
studies
are
summarized
in
Table
16.
Most
compounds
were
stable
for
the
entire
28­
day
period.
However
several
of
the
late
eluters,
including
some
of
the
BDEs
bagan
to
exhibit
reduced
recoveries
at
day
21.
This
trend
was
more
significant
at
day
28.
Sample
hold
times
were
therefore
set
at
14
days.

In
addition
to
investigating
sample­
holding
times,
we
concurrently
ran
an
extract
holding
time
study.
Day­
0
sample
extracts
from
the
holding
time
study
were
analyzed
over
the
same
35­
day
period
to
determine
the
integrity
of
the
extract
when
stored
@
6
°
C.
These
results
are
summarized
in
Table
17.
Extract
hold
times
were
set
at
28
days.

Table
14.
Storage
stability
experimental
design
Exp.
Conditions
Purpose
1
Cincinnati
tap
fortified
with
analytes
at
5
ug/
L
with
standard
method
preservation
and
no
Ohio
River
water
Typical
Cinci.
tap
conditions
that
are
expected
to
perform
well.
These
data
form
the
basis
for
the
method
HT.
2
Cincinnati
tap
fortified
with
analytes
at
5
ug/
L
with
standard
method
preservation
and
with
4
mL/
L
Ohio
River
water
Challenged
Cinci
tap.
Plate
counts
will
be
high
on
day
one
and
then
controlled.
Recoveries
should
be
good.
3
Cincinnati
tap
fortified
with
analytes
at
5
ug/
L
with
standard
method
preservation
but
omitting
the
citric
acid
and
no
Ohio
River
water
Some
compounds
are
more
stable
at
low
pH
and
may
show
some
loss
even
if
microbial
degradation
is
not
active.
Re­
growth
may
occur,
but
degradation
is
not
expected.
4
Cincinnati
tap
fortified
with
analytes
at
5
ug/
L
with
standard
method
preservation
but
omitting
the
citric
acid
and
no
Ohio
River
water
Microbes
will
be
strong
from
day
one
and
will
reach
TNTC
levels.
Recoveries
may
be
diminished,
but
may
be
influenced
by
both
pH
and
microbial
degradation.
22
Table
15.
Sample
holding
time
data
for
target
compounds
fortified
at
5.0
ug/
L
in
chlorinated
surface
water
(
Experiment
1
in
Table
14).

Analyte
Day
0
%
Rec.
Day
7
%
Rec.
Day
14
%
Rec.
Day
21
%
Rec.
Day
28
%
Rec.

Dimethoate
82.5
95.8
88.0
80.9
87.1
Atrazine
78.6
78.9
72.7
72.6
75.5
Propazine
79.1
78.7
74.3
72.1
72.1
Vinclozolin
86.3
83.1
81.7
80.6
78.9
Prometryn
84.7
85.5
84.7
82.9
85.3
Bromacil
96.8
98.5
101
95.4
109
Malathion
86.3
86.1
85.2
79.6
85.5
Chlorpyrifos
84.9
81.8
83.5
74.9
75.3
Thiobencarb
87.1
90.4
86.9
85.1
86.5
Parathion
89.6
87.3
88.1
85.4
92.7
Terbufos­
Sulfone
86.9
89.5
88.7
84.3
86.8
Oxychlordane
80.7
78.9
77.6
75.7
68.8
Esbiol
92.7
93.1
91.3
89.8
88.9
Nitrophen
97.7
94.8
96.4
90.8
96.3
Kepone
68.4
70.8
71.7
74.9
75.7
Norflurazon
103
102
101
99.4
114
Hexazinone
98.5
92.8
116
110
130
Bifenthrin
89.5
80.9
87.6
78.7
72.7
2,2',
4,4'­
Tetrabromodiphenyl
Ether
(
BDE­
47)
87.2
75.6
87.2
76.5
71.0
Mirex
83.8
75.7
84.3
71.8
64.5
2,2',
4,4',
6­
Pentabromodiphenyl
Ether
(
BDE­
100)
87.3
83.5
88.9
78.6
74.5
2,2',
4,4',
5­
Pentabromodiphenyl
Ether
(
BDE­
99)
92.6
77.7
94.1
84.5
78.9
Hexabromobiphenyl
92.5
76.9
92.4
80.5
74.9
Fenvalerate
116
97.7
104
106
97.3
Esfenvalerate
103
84.6
102
96.9
92.4
2,2',
4,4',
5,5'­
Hexabromodiphenyl
Ether
(
BDE­
153)
89.8
79.2
93.3
84.9
87.5
1,3­
Dimethyl­
2­
Nitrobenzene
(
SUR)
74.9
81.9
80.3
79.3
82.4
Triphenylphosphate
(
SUR)
79.9
84.2
87.7
90.8
96.6
Perylene­
d12
(
SUR)
96.9
89.5
92.1
99.1
104
23
Table
16.
Results
for
the
pour
plate
studies
conducted
in
conjunction
with
the
sample
storage
stability
studies.

Day
0
Day
7
Day
14
Day
21
Day
28
Day
35
Experiment
#
Average
Average
Average
Average
Average
Average
CFU/
mL
CFU/
mL
CFU/
mL
CFU/
mL
CFU/
mL
CFU/
mL
#
1
Surface
Water
3
3
0
1
0
0
Preserved/
no
seed
#
2
Surface
Water
44
5
5
2
1
2
Preserved
w/
seed
#
3
Surface
Water
47
11
6
5
4
1
Preserved/
No
citric
acid/
no
seed
#
4
Surface
Water
64
26
10
7
7
7
Preserved/
No
citric
acid/
seed
Table
17.
Results
of
the
extract
storage
stability
studies.

Analyte
Day
0
%
Rec.
Day
7
%
Rec.
Day
14
%
Rec.
Day
21
%
Rec.
Day
28
%
Rec.

Dimethoate
82.5
87.5
84.1
86.9
94.2
Atrazine
78.6
78.1
76.3
78.9
86.1
Propazine
79.1
76.0
77.1
78.7
83.3
Vinclozolin
86.3
85.3
88.8
90.3
87.5
Prometryn
84.7
89.6
86.1
87.1
87.0
Bromacil
96.8
104
98.6
102
112
Malathion
86.3
89.7
87.5
84.4
89.4
Chlorpyrifos
84.9
87.0
87.4
83.7
84.4
Thiobencarb
87.1
92.2
88.7
90.4
88.6
Parathion
89.6
90.4
89.7
90.7
96.2
Terbufos­
Sulfone
86.9
94.4
90.9
90.1
90.5
Oxychlordane
80.7
81.8
83.7
82.3
87.6
Esbiol
92.7
94.2
92.8
96.0
92.9
Nitrophen
97.7
95.8
98.2
97.9
100
Kepone
68.4
73.7
75.3
74.9
75.5
Norflurazon
103
105
106
117
115
Hexazinone
98.5
98.9
122
123
135
Bifenthrin
89.5
88.9
84.5
86.3
87.6
2,2',
4,4'­
Tetrabromodiphenyl
Ether
(
BDE­
47)
87.2
84.2
86.7
81.1
85.7
Mirex
83.8
86.1
81.3
80.5
82.5
2,2',
4,4',
6­
Pentabromodiphenyl
Ether
(
BDE­
100)
87.3
92.2
87.5
85.1
87.0
2,2',
4,4',
5­
Pentabromodiphenyl
Ether
(
BDE­
99)
92.6
90.0
88.2
95.2
95.1
Hexabromobiphenyl
92.5
85.1
89.0
89.7
94.5
Fenvalerate
116
107
101
120
119
Esfenvalerate
103
94.7
102
110
110
24
Analyte
Day
0
%
Rec.
Day
7
%
Rec.
Day
14
%
Rec.
Day
21
%
Rec.
Day
28
%
Rec.

2,2',
4,4',
5,5'­
Hexabromodiphenyl
Ether
(
BDE­
153)
89.8
90.5
84.9
92.3
104
1,3­
Dimethyl­
2­
Nitrobenzene
(
SUR)
74.9
75.1
76.1
77.6
79.9
Triphenylphosphate
(
SUR)
79.9
90.1
88.5
92.9
94.4
Perylene­
d12
(
SUR)
96.9
92.9
89.0
98.5
107
9.0
Evaluation
of
Cartridge
Extraction
Procedure.
Experiments
were
completed
to
determine
method
performance
when
extracting
samples
using
SDVB
cartridges
versus
SDVB
disks.
Precision
and
accuracy
of
extraction
sets
fortified
at
a
final
concentration
of
5
ug/
L
were
evaluated.
Initially
it
was
noticed
during
the
experiments
that
more
water
was
contained
in
the
extraction
eluents
of
the
cartridges
than
was
noticed
in
the
disk
eluents.
As
a
result,
the
drying
step
parameters
used
in
the
method
for
disk
extractions
had
to
be
evaluated
for
cartridges
before
a
proper
comparison
of
the
two
techniques
could
be
accomplished.
Water
in
the
final
extract
has
proven
to
cause
resolution
problems
on
the
GC
and
to
adversely
effect
column
life,
proper
quantitation
of
the
analyte,
and
in
some
cases
analyte
response.
The
first
experiment
assessed
drying
the
eluents
with
5
g
of
sodium
sulfate
in
the
drying
column
versus
drying
the
eluents
with
8.5
g
of
sodium
sulfate.
In
addition,
two
different
types
of
8.5
g
drying
tubes
were
evaluated
because
of
the
concern
that
the
polypropylene
drying
tubes
had
the
potential
to
adversely
effect
method
performance.
One
was
a
polypropylene
tube
with
a
frit
and
one
was
a
disposable
10­
mL
glass
pipette
with
a
glass
wool
plug.
In
each
case
the
experiment
involved
directly
fortifying
a
volume
of
extract
that
contained
a
small
amount
of
water
(
0.4
mL)
and
then
processing
the
extract
with
the
target
analytes
to
isolate
the
effect
of
the
drying
step
only.

Results
from
the
drying
tube
evaluation
can
be
found
in
Table
18.
As
can
be
seen
from
the
data,
there
seems
to
be
no
difference
between
the
two
drying
tubes.
Additionally
the
data
indicates
that
the
5
g
drying
tube
is
still
adequate
for
drying
the
eluents
for
cartridge
extractions.
One
concern
remains
however,
the
fact
that
water
amounts
in
the
eluents
will
vary
from
extraction
to
extraction.
It
will
be
up
to
the
discretion
of
the
analyst
to
determine
if
there
are
copious
amounts
of
water
in
the
eluents
(>
2
mL)
and
to
use
a
larger
drying
column
when
necessary.
This
study
proves
that
up
to
8.5
g
of
sodium
sulfate
can
be
used
to
dry
the
eluents
if
necessary
without
adversely
effecting
analyte
recovery.
It
should
be
noted
that
the
larger
drying
columns
were
eluted
with
two
times
the
amount
of
solvent
to
make
up
for
the
roughly
2­
fold
increase
in
the
sodium
sulfate
bed.

Next,
data
from
fortified
reagent
waters
(
containing
all
Method
527
reagents)
were
extracted
with
cartridges
and
with
disks
(
Table
19.).
Lower
percent
recoveries
were
observed
for
the
cartridge
data
than
from
samples
extracted
with
disks.
This
was
especially
true
for
the
heavier
molecular
weight
compounds.
Earlier
experiments
proved
that
drying
bed
size,
excess
water
in
the
eluents,
and
increased
solvent
elution
volumes
did
not
effect
analyte
recoveries
under
the
proper
conditions.
As
a
result
we
concluded
that
that
the
lower
recoveries
must
be
associated
with
incomplete
adsorption
by
the
cartridge
SDVB.
Therefore,
cartridges
are
not
allowed
to
be
used
in
the
method.
25
Table
18.
The
effect
of
the
drying
step
and
column
type
on
compound
recovery.

Analyte
Begin
check
Mid
Check
End
Check
5g
Control
8.5g
glass
8.5g
plastic
%
rec
%
rec
%
rec
Ave
%
Rec
Ave
%
Rec
Ave
%
Rec
1,3­
dimethyl­
2­
nitrobenzene
(
surr)
104
103
103
89
88
89
Dimethoate
96
92
92
90
86
86
Atrazine
99
101
99
95
92
91
Propazine
97
97
96
92
89
89
Vinclozolin
95
97
99
91
90
90
Prometryn
93
94
96
90
87
89
Bromacil
94
92
95
89
85
87
Malathion
91
96
98
90
87
89
Thiazopyr
90
92
97
89
87
91
Chlorpyrifos
95
96
99
91
92
92
Thiobencarb
94
99
96
92
89
90
Parathion
95
94
98
93
90
90
Terbufos­
sulfone
93
95
96
90
86
88
Oxychlordane
92
96
97
92
91
93
Esbiol
95
97
95
92
91
90
Fenamiphos
100
95
99
114
105
90
Nitrophen
100
97
100
98
94
96
Kepone
91
99
101
101
98
101
Norflurazon
96
98
98
91
88
88
Hexazinone
92
96
97
89
87
89
Triphenylphosphate
(
surr)
95
104
103
100
99
101
Bifenthrin
94
97
100
88
85
87
BDE­
47
90
91
96
87
86
88
Mirex
87
94
93
86
86
89
BDE­
100
91
95
96
86
84
87
BDE­
99
99
98
98
89
87
89
Perylene­
d12
(
surr)
94
98
102
86
82
85
Hexabromobiphenyl
90
93
101
88
82
86
Fenvalerate
85
91
95
75
72
73
Esfenvalerate
89
90
96
79
75
77
BDE­
153
91
96
104
91
83
93
Average
Recovery
94
96
98
91
88
89
26
Table
19.
Comparison
of
disk
and
cartridge
extraction
recoveries.

Disk
Cartridge
Disk
Cartridge
Analyte
5.0
ug/
L
5.0
ug/
L
5.0
ug/
L
5.0
ug/
L
%
rec
%
rec
%
rsd
%
rsd
1,3­
dimethyl­
2­
nitrobenzene
(
surr)
87
88
3.1
2.7
Dimethoate
60
106
3.0
2.3
Atrazine
93
96
3.8
2.5
Propazine
87
91
4.9
3.5
Vinclozolin
94
94
2.3
2.5
Prometryn
98
100
3.8
2.8
Bromacil
101
106
2.2
0.9
Malathion
90
95
2.2
2.4
Thiazopyr
87
88
2.9
2.2
Chlorpyrifos
89
80
2.3
3.6
Thiobencarb
89
91
2.0
3.0
Parathion
96
98
3.4
1.5
Terbufos­
sulfone
95
99
2.5
1.4
Oxychlordane
85
72
4.9
3.7
Esbiol
94
81
3.0
3.5
Fenamiphos
128
118
2.9
6.0
Nitrophen
106
105
3.4
2.2
Kepone
95
94
4.0
3.0
Norflurazon
113
115
3.7
0.6
Hexazinone
103
105
3.1
2.4
Triphenylphosphate(
surr)
104
105
2.7
2.0
Bifenthrin
86
71
3.3
4.6
BDE­
47
85
68
3.8
7.1
Mirex
80
63
3.2
6.0
BDE­
100
83
69
5.8
4.6
BDE­
99
84
67
3.0
6.5
Perylene­
d12
(
surr)
86
80
3.6
2.0
Hexabromobiphenyl
89
69
2.5
6.9
Fenvalerate
97
78
1.8
6.7
Esfenvalerate
91
73
1.8
5.9
BDE­
153
87
68
2.5
8.3