Document ID: EPA-HQ-OPP-2015-0197-0002
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2015-05-20T04:00Z

EPA Registration Division contact: Susan Lewis, (703) 305-7090

  SEQ CHAPTER \h \r 1 Interregional Research Project Number 4 (IR-4)

5E8349

	EPA has received a pesticide petition (5E8349) from IR-4, 500 College
Road East, Suite 201W, Princeton, New Jersey, 08540, proposing, pursuant
to section 408(d) of the Federal Food, Drug, and Cosmetic Act (FFDCA),
21 U.S.C. 346a(d), to amend 40 CFR part 180 by establishing a tolerance
for residues of fluazinam, including its metabolites and degradates, in
or on the raw agricultural commodity mayhaw at 2.0 parts per million
(ppm), cabbage at 3.0 ppm, cucurbit vegetables, squash/cucumber subgroup
9B at 0.05 ppm, vegetable, brassica leafy, group 5, except cabbage at
0.01 ppm, and vegetable, tuberous and corm, subgroup 1C at 0.02 ppm. 
Upon approval of the aforementioned tolerances, IR-4 requests to remove
the established tolerances for fluazinam on vegetable, brassica leafy,
group 5 of 0.01 ppm and potato at 0.02 ppm.  EPA has determined that the
petition contains data or information regarding the elements set forth
in section 408 (d)(2) of  FDDCA; however, EPA has not fully evaluated
the sufficiency of the submitted data at this time or whether the data
supports granting of the petition. Additional data may be needed before
EPA rules on the petition. 

A. Residue Chemistry

Plant metabolism.   SEQ CHAPTER \h \r 1 The residue of concern is best
defined as the parent, fluazinam in most crops and as the parent,
fluazinam and its metabolite, AMGT, in fruit crops (grape and
blueberry).    SEQ CHAPTER \h \r 1 The metabolism of fluazinam in plants
(potatoes, peanuts, apples and wine grapes) is adequately understood for
the purposes of these tolerances.  The metabolism of fluazinam in peanut
and potato involves initial reduction of the nitro groups, hydrolysis of
the trifluoromethyl group as well as replacement of chlorine by
glutathione with subsequent reactions along the glutathione pathway. 
Following replacement of the deactivating NO2 and Cl groups with
activating groups such as OH, NH2 and sulfhydryl, ring cleavage occurs. 
Parent fluazinam is then rapidly degraded to form CO2 and carbon
fragments which are incorporated into natural products such as glucose,
fructose, sucrose, oils and protein.  Thus, parent fluazinam is either
not found or barely detectable in peanuts and potatoes.  Fluazinam
parent was the major identifiable residue in both the grape and apple
metabolism studies.  However, minor levels of AMGT (less than 5% of the
total radioactive residue) were also formed in grapes and apples.  In
grape and apple metabolism studies, following reduction of the nitro
groups and replacement of chlorine with a sulfur containing side-chain
such as glutathione (as occurred in peanuts and potatoes), glucose is
attached to the thiolactic acid conjugate of AMPA to form the metabolite
known as AMGT.  This metabolite was not found in   SEQ CHAPTER \h \r 1
the peanut or potatoes metabolism studies.  It is analogous to the
cysteine conjugate of AMPA found in rats.  Identifiable residues in
plant metabolism studies either closely resemble fluazinam in structure
or are the result of re-incorporation of the fluazinam carbon pool into
natural products. 

    Ruminant and poultry metabolism studies demonstrated that the
transmittal of residues from the feed of goats and hens through to meat,
milk, and eggs was low.  Total 14C residues were below 1 ppm in all
tissues, milk and eggs.  Identifiable residues were less than 2% of the
administered dose in all matrices, except for chicken fat and liver. 
The major metabolite found in animal tissues and milk were DAPA, AMPA,
and their sulfamate conjugates.  Parent Fluazinam was only detected in
very small amounts.

	2. Analytical method. An analytical method using LC-MS/MS for the
determination of fluazinam and AMGT residues on cabbage, squash and
cucumbers has been developed and validated.  The method involves solvent
extraction followed by liquid-liquid partitioning and concentration
prior to a final purification.  The method has been successfully
validated by an independent laboratory using peanut nutmeat as the
matrix.  The limit of quantitation (LOQ) of the method is 0.01 ppm for
both fluazinam and AMGT in these crops.

	3. Magnitude of residues. Cabbage. A total of 8 field trials were
conducted on cabbage.  Treatment 2 of each field site consisted of one
application applied as 0.055 lb. a.i./1000 plants in a plant base drench
at the time of transplanting followed by six (6) foliar applications at
the target rate of approximately 0.5 lb. a.i./A, applied at 7 (+1) day
intervals.  Cabbages were harvested approximately 7 days after the last
application.   One decline trial was included with sampling at 0, 3, 7,
9, 14 and 21 days after the last application.  The results show that the
maximum Fluazinam residue in cabbage following a total foliar
application of approximately 3 lb. a.i./A/season with an average PHI of
7 days, was 1.7 ppm.  The mean fluazinam residue was 0.64 ppm. Residues
generally declined over time.  Only one residue of AMGT above the LOQ
(0.01 ppm) at 0.011 ppm was observed.  

Squash. A total of 6 field trials were conducted on summer squash. 
Treatment 2 of each field site consisted of one soil directed and four
(4) foliar applications at the target rate of approximately 0.78 lb.
a.i./A, applied at 7 (+1) day intervals.  Squash were harvested 6 to 7
days after the last application.  The results show that the maximum
Fluazinam residue in summer squash following a total application of
approximately 3.9 lb a.i./A/season with an average PHI of 7 days, was
0.016 ppm.  The mean fluazinam residue was 0.011 ppm.  No residues of
AMGT above the LOQ of 0.01 ppm were observed in any sample.  

Cucumber.  A total of 6 field trials were conducted on cucumbers. 
Treatment 2 of each field site consisted of one soil directed at
transplanting and four (4) foliar applications at the target rate of
approximately 0.78 lb. a.i./A, applied at 7 (+2) day intervals, for a
total of 3.9 lb. a.i./A.  (Two sites received more than the target
amount: 4.6 and 5.7 lb. a.i./A.)  Cucumbers were harvested 6 to 7 days
after the last application.  The results show that the maximum Fluazinam
residue in cucumbers following a total application of approximately 3.9
– 5.7 lb a.i./A/season with an average PHI of 7 days, was 0.032 ppm. 
The mean fluazinam residue was 0.013 ppm.  No residues of AMGT above the
LOQ of 0.01 ppm were observed in any sample.  

B. Toxicological Profile

	1. Acute toxicity.  A battery of acute toxicity studies was conducted
which placed technical fluazinam in Toxicity Category IV for acute oral
LD50 and dermal irritation, and Category III for dermal LD50, inhalation
LC50 and eye irritation.  Technical fluazinam showed potential for
dermal sensitization.  In an acute neurotoxicity study, the NOAEL for
neurotoxicity was 2000 mg/kg bw (the highest dose tested) and the NOAEL
for systemic effects was 50 mg/kg bw.

	2. Genotoxicty. A battery of tests has been conducted to assess the
genotoxic potential of technical fluazinam.  Assays conducted included
two gene mutation tests in bacteria, a chromosomal aberration test in
mammalian cells, a mouse micronucleus test and a DNA repair test in
bacteria.  Technical fluazinam did not elicit a genotoxic response in
any of the studies conducted.

	3. Reproductive and developmental toxicity. In a two-generation
reproductive toxicity study, the NOAEL for reproductive effects was 100
ppm (10.6 mg/kg bw/day).  The NOAEL for parental toxicity was 20 ppm
(1.9 mg/kg bw/day).

    In a rat developmental study, there were no developmental effects
observed at non-maternally toxic doses.  The developmental NOAEL was 50
mg/kg bw/day and the LOAEL was 250 mg/kg bw/day, based upon decreased
mean fetal body weight and other evidence suggestive of delayed fetal
development related to maternal toxicity. The maternal NOAEL was shown
to be 50 mg/kg bw/day. 

    In a rabbit developmental study, there were no developmental effects
observed at non-maternally toxic doses.  The developmental NOAEL was 7
mg/kg bw/day and the LOAEL was 12 mg/kg bw/day, based on increased
incidence of total litter loss and possible slightly increased
incidences of fetal findings at this dose.  It was concluded that the
maternal NOAEL was 4 mg/kg bw/day.

	4. Subchronic toxicity. The NOAEL for the 13-week feeding study in rats
was 50 ppm (3.8 mg/kg bw/day in males, 4.3 mg/kg bw/day in females). 
The LOAEL was 500 ppm (38 mg/kg bw/day in males, 44 mg/kg bw/day in
females), based on periacinar hepatocellular hypertrophy and sinusoidal
chronic inflammation in males, increased liver weights in males and
increased lung weights in females.

    In a 13-week dog study, the NOAEL was 10 mg/kg bw/day.  The LOAEL
was 100 mg/kg bw/day, based on ocular change observed
ophthalmoscopically and liver effects consisting of increased relative
liver to body weight, bile duct hyperplasia with or without
cholangiofibrosis and increased plasma phosphatase levels.

    In a 21-day dermal study, the NOAEL for systemic effects was 10
mg/kg bw/day.  The LOAEL was 100 mg/kg bw/day, based on hepatocelluar
hypertrophy and increases in AST and cholesterol levels. 

    In a subchronic neurotoxicity study, no effects considered to be
indicative of neurotoxicity were observed at the highest dose tested,
3000 ppm (233 mg/kg bw/day in males, 280 mg/kg bw/day in females).  The
NOAEL for systemic toxicity (body weight differences) was 1000 ppm (74
mg/kg bw/day). 

    In a developmental neurotoxicity study in rats, fluazinam was
administered by gavage to female rats from Day 6 after mating to Day 20
of lactation and additionally to their offspring from Day 7 of age to
Day 20 or 21 of age.  The author concluded that the maternal NOAEL was 2
mg/kg bw/day based on reduced body weights and lower food intake.  No
effects were seen on the microscopic structure of the nervous system of
the dams at any dose level (maximum dose of 50 mg/kg bw/day).  The NOAEL
for behavior and nervous system of the dams was >50 mg/kg bw/day, the
highest dose tested.  No adverse effect of treatment at any dose level
was seen on number of implantations, litter size or offspring survival. 
Signs of general toxicity in the offspring were evident based on lower
Day 1 body weights and lower weight gains at 10 and 50 mg/kg bw/day
through weaning.  The NOAEL for general toxicity to offspring was 2
mg/kg bw/day.  There was no evidence of developmental neurotoxicity in
the offspring.  The NOAEL for the functional and morphological
development of the nervous system in the offspring was >50 mg/kg bw/day,
the highest dose tested.  There was no increased sensitivity of the
fetus or young rat pups to fluazinam as compared to the dams.

	5. Chronic toxicity. Fluazinam was not carcinogenic in rats.  A NOAEL
of 10 ppm (Males: 0.38 mg/kg bw/day; Females: 0.47 mg/kg bw/day) of
fluazinam was established based on the following effects at 1000 and/or
100 ppm: lower food consumption and efficiency of food utilization,
slight anemia, elevated cholesterol, increased liver weights, an
increased number of macroscopic liver and testes lesions and an
increased incidence of microscopically observed lung, liver, pancreas,
lymph node and testes lesions.

    An additional study was conducted to further define the NOAEL for
long-term effects in the rat.  In the second study, a NOAEL of 50 ppm
(2.2 mg/kg bw/day) was established based on liver and testes effects.

    Two long-term feeding studies were conducted in mice.  In the first,
the NOAEL for all effects was 10 ppm (Males: 1.1 mg/kg bw/day; Females:
1.2 mg/kg bw/day) and the LOAEL was 100 ppm (Males: 10.7 mg/kg bw/day;
Females: 11.7 mg/kg bw/day) based on the treatment-related effects
observed in the liver.

    A second oncogenicity study in mice was conducted at 1000, 3000 and
7000 ppm to ensure that an MTD dose was studied.  Findings included
increased female mortality, reduced body weight gains, increased brain
weights and/or liver weights.  An impurity in the test material used in
this study resulted in vacuolation of the white matter of the brain and
cervical spinal cord in treated animals. A statistically significant
higher incidence of hepatocellular adenomas was observed in the 3000 ppm
dose males.  Hepatocellular adenomas are common tumors in male mice. 
There was no dose relationship in the induction of the adenoma and no
increase in hepatocellular carcinomas.  It was concluded that fluazinam
is not carcinogenic in the mouse. 

    In a chronic dog study, the NOAEL was determined to be 1 mg/kg
bw/day.  The LOAEL was 10 mg/kg bw/day based on generalized, nonspecific
toxicity.  No ocular effects were observed ophthalmoscopally at any dose
in this study.

	6. Animal metabolism. After an oral dose of fluazinam the median peak
time for blood concentration of radiolabel activity for both sexes was 6
hours.  The major route of excretion was the feces with urine
contributing as a minor route. Less than 1% of the administered dose was
found in the terminated animals.  The highest concentration in tissues
was found in the liver.  There were no major differences related to sex
or dose level in the findings.  It was concluded that fluazinam is
metabolized by both reduction and glutathione and glucuronide
conjugation and further metabolism.

	7. Metabolite toxicology. The same metabolic processes occur in plants
and animals but metabolism in plants is more extensive than in animals.
All of the major identified metabolites in both plants and animals
retain the phenylpyridinylamine structure.  Many of the metabolites
resulting from fluazinam are similar in plants and animals and,
therefore, have already been evaluated toxicologically.  

    Because of the rapid and complete elimination (in animals) and
re-incorporation (in plants) of fluazinam, the toxicity of metabolites
is expected to be similar to but lower than the toxicity of the parent
compound.  The residue of concern is parent fluazinam only in most crops
and parent fluazinam plus its minor metabolite AMGT on/in fruit crops
such as grape and blueberry.

	8. Endocrine disruption. The toxicological profile of fluazinam shows
no evidence of physiological effects characteristic of the disruption of
the hormone estrogen in mammalian chronic studies or in mammalian or
avian reproduction studies.   It is therefore considered that there is
an adequate level of safety over the reference dose for possible
endocrine effects and that an additional safety factor for possible
endocrine effects is not warranted.

C. Aggregate Exposure

	1. Dietary exposure. Potential dietary exposures from food were
estimated using the proposed tolerances for all crops, using the Dietary
Exposure Evaluation Model-Food Consumption Intake Database
(DEEM-FCIDTM)/Calendex Version 4.02/10.00 Release and percent crop
treated of 100%.  The following raw agricultural commodities were
included: apple; carrot; brassica (cole) leafy vegetables (Group 5)
including turnip greens and cabbage at the proposed increased tolerance
level; bean (Groups 6A, 6B and 6C); bushberry (Group 13-07B); ginseng;
head lettuce; leaf lettuce; bulb onion (Crop 3-07A); peanut; vegetable,
tuberous and corm, subgroup 1C; pepper/eggplant subgroup 8-10B; 
cucurbit vegetables, melon subgroup 9A and squash/cucumber subgroup 9B;
soybeans; imported wine grapes and sherry; and resulting secondary
residues in meat and milk.  For acute dietary exposure, the acute
population adjusted dose (aPAD) was based on the NOAEL of 50 mg/kg
bw/day from an acute rat neurotoxicity study.   For females 13-49 years
of age, the NOAEL of 7 mg/kg bw/day from the rabbit developmental
toxicity study was used for acute dietary exposure.  For chronic dietary
exposure, the chronic population adjusted dose (cPAD) was based on the
NOAEL from the mouse carcinogenicity study (1.1 mg/kg bw/day).  An
uncertainty factor of 100 was used in both cases since the results of a
DNT study confirmed there was no increased sensitivity of the fetus or
young rat pups to fluazinam as compared to the dams and the FQPA
uncertainty factor could be reduced to one (1).

	i. Food. Acute Risk Tier 1 acute dietary exposure analyses were
conducted for fluazinam to determine the exposure contribution of the
above mentioned commodities to the diet and to ascertain the acute risk
potential.  The estimates were based on: proposed tolerance level
residues for all the crops; apple, peanut and potato processing studies;
market share assumptions of 100% crop treated; screening-level acute
EDWC of 218 ppb for water; and consumption data from NHANES/WWEIA
(2005-6, 2007-8, and 2009-10 cycles) continuing survey of food intake.

    Even using all of the worst-case exposure scenarios listed above,
the Tier 1 95th percentile acute dietary exposure (per capita) from food
and drinking water for the U.S. population was estimated to be 0.026340
mg/kg bw/day or 5.27% of the aPAD. The highest acute exposure estimate
(95th percentile) was observed in the females 13-49 yrs. subpopulation:
0.019739 mg/kg bw/day.  This corresponds to 28.2% of the aPAD. 

    Chronic Risk Tier 1 dietary exposure analyses were conducted for
fluazinam to determine the exposure contribution of the above mentioned
commodities to the diet and to ascertain the chronic risk potential. The
estimates were based on: proposed tolerance level residues for all the
crops; apple, peanut and potato processing studies; market share
assumptions of 100% crop treated; screening-level chronic EDWC of 37
ppb; and consumption data from NHANES/WWEIA (2005-6, 2007-8, and 2009-10
cycles) continuing survey of food intake.

    Even using all of the worst-case exposure scenarios listed above,
the Tier 1 chronic dietary exposure estimates resulted in an estimated
exposure for the U.S. population of 0.002375 mg/kg bw/day.  This
exposure corresponds to 21.6% of the cPAD of 0.011mg/kg bw/day.  The
highest exposure estimate was calculated for the non-nursing infants
population subgroup.  This exposure was determined to be 0.007247 mg/kg
bw/day (65.9% of the cPAD) most of which was due to the water
contribution.

    It can be concluded that acute or long-term dietary exposure to
fluazinam through residues on treated apple, carrot, brassica (cole)
leafy vegetables (Group 5) including turnip greens, bean (Groups 6A, 6B
and 6C), bushberry (Group 13-07B), ginseng, head lettuce, leaf lettuce,
bulb onion (Crop 3-07A), peanut, tuberous and corm, subgroup 1C,
pepper/eggplant subgroup 8-10B,  cucurbit vegetables, melon subgroup 9A
and squash/cucumber subgroup 9B, soybeans, imported wine grapes and
sherry and resulting secondary residues in meat and milk should not be
of cause for concern.

	ii. Drinking water. Since fluazinam is intended for application
outdoors to field grown crops, the potential exists for parent and/or
metabolites to reach ground or surface water that may be used for
drinking water.  The potential estimated drinking water concentrations
(EDWCs) associated with the application of fluazinam to cabbage, mayhaw,
squash/cucumber subgroup 9B, or tuberous and corm, subgroup 1C, is less
than the maximum values estimated for turf.  Thus, the more conservative
screening-level acute EDWC (218 ppb) and chronic EDWC (37 ppb) for turf
were incorporated into the DEEM-FCID model to estimate dietary exposure.

	2. Non-dietary exposure. A registration for the use of fluazinam on
golf courses has been approved.  Applications are to be made by golf
course workers only, thus no residential handler exposure is expected. 
However, post-application dermal exposure from contact with the treated
turf on golf courses may occur.  Thus, post-application dermal exposure
was calculated for adults and for youth playing golf, using the general
US population subgroup and youth, 13-19 years subgroup.   The Margins of
Exposure (MOE) for both the adults (21,800) and the youth (14,300) were
well above the acceptable MOE of 100.  

D. Cumulative Effects

	Fluazinam is a phenylpyridinylamine fungicide.   Since there are no
other members of this class of fungicides, it is considered unlikely
that fluazinam would have a common mechanism of toxicity with any other
pesticide in use at this time.

E. Safety Determination

	1. U.S. population. Dietary and post-application dermal exposure from
recreational contact with treated turf on golf courses will be the
routes of exposure to the U.S. population.  Ample margins of safety have
been demonstrated for both situations. For the general U.S. population,
the chronic aggregate risk assessment is based on exposure from food and
drinking water only, since no residential uses exist.  As noted above,
the chronic dietary exposure to fluazinam represents 21.6% of the cPAD
for the overall U.S. population, using the conservative Tier I
assumptions. A cancer dietary assessment was not conducted because this
product has been classified as “suggestive evidence of
carcinogenicity, but not sufficient to assess human carcinogenic
potential.”

An acute aggregate risk assessment is required for the general
population and for the subgroup of females 13-49 years old.  For both of
these populations, the acute dietary (food and drinking water) risk
assessment represents acute aggregate risk since the dietary route alone
is relevant for acute exposure and risk assessment. As noted above, the
acute dietary exposure to fluazinam is estimated to be 5.27% of the aPAD
for the overall U.S. population, and 28.2% of the aPAD for the subgroup
of females 13-49 years old. These values were presented above and are
below the Agency’s level of concern. 

Short-term and intermediate-term aggregate risks include chronic dietary
and short-term residential sources of exposure. Since Fluazinam is
proposed for use on golf courses there is potential for post-application
exposure to adults and youths from playing golf.  The US general
population was chosen as the representative short-term population for
adults, while the dietary subpopulation youths aged 13 to 19 was chosen
to match with the youth golfer residential exposure.  The dietary (food
and drinking water) exposure was based on the Tier I chronic exposure
assessment as noted above but converted to an MOE.  This was aggregated
with the appropriate post-application residential golfer MOE to yield
the following aggregate short/intermediate-term MOEs: US general
population MOE = 1811; youths, 13-19 = 2779.  These aggregate risks are
well above 100 for all populations, indicating acceptable risks for both
adults and youth.  

Based on this information, it can be concluded that there is reasonable
certainty that no harm will result from acute, short/intermediate term
or chronic exposure to fluazinam.

	2. Infants and children. Data from developmental toxicity studies in
the rat and rabbit, a 2-generation reproduction study and a
developmental neurotoxicity study were considered.  These studies, that
were described earlier, demonstrated no increased sensitivity of rats or
rabbits to in utero or gavage exposure of pups to fluazinam.  In
addition, the multigeneration reproductive toxicity study did not
identify any increased sensitivity of rats to in utero or postnatal
exposure.  For all four studies, parental NOAELs were lower than or
equivalent to the developmental or offspring NOAELs.  It is concluded
that the standard margin of safety will protect the safety of infants
and children and that an additional FQPA safety factor is not warranted.

    The dietary exposure of fluazinam to infants and children is
estimated to be less than any level of concern. The proposed tolerances
plus drinking water input will utilize 20.3% of the aPAD for the
“Children (1-2 years)” subgroup, and 53.6% of the cPAD for the
“Children (1-2 years)” subgroup.  The non-nursing infants population
subgroup utilizes 65.9% of the cPAD.  (These were the most sensitive
infant/child subgroups.) Thus, it can be concluded that there is
reasonable certainty that no harm will result to infants and children
from acute or chronic exposure to fluazinam.

F. International Tolerances

	There are presently no Codex maximum residue levels (MRLs) established
for residues of fluazinam on any crop.  The Canadian established MRLs
for brassicas (crop group 5A & 5B), bushberries (crop group 13B), dry
beans, lima beans, snap beans, and the Mexican MRLs for beans and
potatoes are similar to the tolerances established for these crops in
the USA.

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