EPA Registration Division contact: Andrew Ertman, 703-308-9367

Interregional Research Project Number 4 (IR-4)

PP No. 1E7959

EPA has received a pesticide petition (PP No. 1E7959) 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 Cosme tic Act (FFDCA), 21 U.S.C. 346a(d), to amend 40 CFR part 180 by establishi ng a tolerance for residues of fluazinam in or on the raw agricultural commodity fruiting vegetables group, pepper/eggplant subgroup

8-10B at 0.10 parts per million (ppm) and cucurbit vegetables, melon subgroup 9A at 0.08 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.

This notice includes information from a separate petition submitted by ISK Biosciences Corporation, 7470 Aubur n Road, Suite A, Concord, OH 44057.

A. Residue Chemistry

Plant metabolism. 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). 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 degrad ed to form CO2 and carbon fragments which are incorporated into natural products such as glucose, fr uctose, 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. Howe ver, 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, followi ng reduction of the nitro groups and replacement of chlorine with a sulfur cont aining 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 the peanut or potatoes metabo lism 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 ca rbon pool into natural products. Ruminant and poultry metabolism studi es demonstrated that the transmittal of residues from the feed of goats and hens th rough to meat, milk, and eggs was low. Total 14C residues were below 1 ppm in all tissues, milk and eggs. Identifiable2

residues were less than 2% of the admini stered dose in all matrices, except for chicken fat and liver. The major metaboli te found in animal ti ssues and milk were DAPA, AMPA, and their sulfamate conj ugates. 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 cantaloupe and pepper has been developed and validated. The method in volves solvent extraction followed by liquid-liquid partitioning and concentratio n prior to a final purification. The method has been successfully validated by an independent laboratory using peanut nutmeat as the matrix. The limit of qua ntitation (LOQ) of the method is 0.01 ppm for both fluazinam and AMGT in both crops.

3. Magnitude of residues. Cantaloupe. A total of 11 field trials were conducted on cantaloupe. Treatment 2 of each field si te consisted of six (6) foliar applications at the target rate of approximately 0.78 lb a.i./A, applied at 7 (+1) day intervals. Cantaloupes were harvested approximately 30 days after the last application. One

decline trial was included with sampling at 6, 12, 19, 28 and 34 days after the last application. The results show that the maximum Fluazinam residue in cantaloupes following a total application of approximate ly 4.68 lb a.i./A/season with an average PHI of 30 days, was 0.077 ppm. The mean fluazinam residue was 0.014 ppm.

Residues declined over time. No resi dues of AMGT above the LOQ of 0.01 ppm were observed in any sample.

Pepper. A total of 13 field trials were conduc ted on pepper (9 on bell and 4 on nonbell). Treatment 2 of each field site consiste d of six (6) applications at the target rate of approximately 0.78 lb a.i./A, applied in general at 7 day intervals but was extended between the third and fourth applicat ion at some sites to achieve the target PHI. Peppers were harvested approximately 30 days after the last application. One decline trial was included with samplin g at 7, 13, 20 and 35 days after the last application. The results show that th e maximum Fluazinam residue following a total application of approximately 4.68 lb a.i ./A/season with an average PHI of 30 days, was 0.038 ppm in bell peppers and 0.053 ppm in non-bell peppers. The mean fluazinam residue was 0.011 ppm in bell peppers and 0.017 ppm in non-bell peppers. Residues declined over time. 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 Ca tegory IV for acute oral LD50 and dermal irritation, and Category III for dermal LD 50, inhalation LC50 and eye irritation. Technical fluazinam showed potential for dermal sensitization. In an acute neurotoxicity study, the NOAEL for neurot oxicity was 2000 mg/kg bw (the highest dose tested) and the NOAEL for systemic effects was 50 mg/kg bw.3

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 reproduc tive effects was 100 ppm (10.6 mg/kg bw/day). The NOAEL for parental toxi city was 20 ppm (1.9 mg/kg bw/day). In a rat developmental study, there w ere no developmental effects observed at non-maternally toxic doses. The deve lopmental 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 in creased 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 a nd sinusoidal chronic inflammation in males, increased liver weights in males and increased lung weights in females. In a 13-week dog study, the NOA EL 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 li ver 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 high est dose tested, 3000 ppm (233 mg/kg bw/day in males, 280 mg/kg bw/day in females). The NOAEL for system ic 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 aft er 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 an y dose level (maximum dose of 50 mg/kg bw/day). The NOAEL for behavior and ner vous 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 th rough weaning. The NOAEL for general4

toxicity to offspring was 2 mg/kg bw/day. There was no evidence of developmental neurotoxicity in the offspring. The NOA EL 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 carcinogeni c 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 effe cts 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 stud y, 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 condu cted 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-relat ed 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, increa sed 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 inducti on of the adenoma and no increase in hepatocellular carcinomas. It was conclude d 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 generaliz ed, nonspecific toxicity. No ocular effects were observed ophthalmoscopally at any dose in this study.

6. Animal metabolism. After an oral dose of flua zinam the median peak time

for blood concentration of radiolabel activi ty 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 th e liver. There were no major differences related to sex or dose level in the finding s. It was concluded that fluazinam is metabolized by both reduction and glut athione and glucuronide conjugation and

further metabolism.

7. Metabolite toxicology

. The same metabolic processes occur in plants and animals but metabolism in plants is more ex tensive than in animal s. All of the major identified metabolites in both plants a nd animals retain the phenylpyridinylamine structure. Many of the metabolites resulti ng from fluazinam are similar in plants5

and animals and, therefore, have already been evaluated toxicologically. Because of the rapid and complete e limination (in animals) and re-incorporation (in plants) of fluazinam, the toxicity of me tabolites is expected to be similar to but lower than the toxicity of the parent co mpound. 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 characteri stic of the disruption of the hormone estrogen in mammalian chronic studies or in mammalian or avian reproduction

studies. It is therefore cons idered that there is an adequa te 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 (except apples as noted below), using the Dietary Exposure Evaluation Model-Food Consumption Intake Database (DEEMFCID TM ) and percent crop treated of 100% (with the exception of imported wine grapes as discussed below). The following raw agricultural commodities were included: 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, potato, pepper/eggplant subgroup 8-10B, melon subgroup 9A, soybea ns, imported wine grapes and sherry and resulting secondary residues in meat an d 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 st udy. 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 wa s 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 exposu re contribution of the above mentioned

commodities to the diet and to ascertain th e 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 (except wine grapes where it was assumed that 100% of the imported wine grapes would be

treated and 39% of wine consumed in th e US is imported); screening-level acute EDWC of 218 ppb for water; and consum ption data from USDA’s CSFII (1994 through 1998) continuing survey of food intake. Even using all of the worst-case ex posure scenarios listed above, the Tier 1 95th percentile acute dietary exposure (per capita ) from food and drinking water for the6

U.S. population was estimated to be 0.024402 mg/kg bw/day or 4.9% of the aPAD. The highest acute exposure estimate (95th percentile) was observed in the females 13-49 yrs. subpopulation: 0.018334 mg/kg bw /day. This corresponds to 26.1% of

the aPAD. Chronic Risk Tier 1 dietary exposu re analyses were conducted for fluazinam to determine the exposure contribution of the above mentioned commodities to the diet and to ascertain the chronic risk potentia l. The estimates were based on: proposed tolerance level residues for all the crops, accept apples where av erage field residues were used; apple, peanut and potato pro cessing studies; market share assumptions of 100% crop treated (except wine grapes where it was assumed that 100% of the imported wine grapes would be treated a nd 39% of wine consumed in the US is imported); screening-level ch ronic EDWC of 37 ppb; and consumption data from USDA’s CSFII (1994 through 1998) cont inuing 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 0.001924 mg/kg bw/day. This exposure corresponds to 17.5% of the cPAD of 0.011mg/kg bw/day. The highest exposure estimate was calculated for the all infants (<1 year) population subgroup. This exposure was determined to be 0.005609 mg/kg bw/day (51.0% 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, br assica (cole) leafy vegetables (Group 5) including turnip greens, bean (Groups 6A, 6B and 6C), bushberry (Group 13-07B), ginseng, head lettuce, leaf lettuce, bul b onion (Crop 3-07A), peanut, potato, pepper/eggplant subgroup 8-10B, melon subgroup 9A, soybeans, imported wine grapes and sherry 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 dr inking water. The potential estimated drinking water concentrations (EDWCs) associated with the application of fluazinam to pepper/eggplant subgroup 8-10B, melon subgroup 9A, or soybeans, 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 is pending. 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 turfon golf courses may occur. Thus, postapplication 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 th e adults (21,800) and the youth (14,300) were well above the acceptable MOE of 100.

D. Cumulative Effects7

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-applicatio n 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 17.5% of the cPAD for the overall U.S. population, using the conservative Tier I assumptions. A cancer dietary assessmen t was not conducted because this product has been classified as “suggestive eviden ce of carcinogenicity, but not sufficient to assess human carcinogenic potential.”

An acute aggregate risk assessment is re quired 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 fo r acute exposure and risk assessment. As noted above, the acute dietary exposure to fl uazinam is estimated to be 4.9% of the aPAD for the overall U.S. population, and 26.1% of the aPAD for the subgroup of females 13-49 years old. These values were presented above and are below the Agency’ level of concern.

Short-term and intermediate-term aggregat e 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-appl ication exposure to adults and youths from playing golf. The US general popul ation was chosen as the representative short-term population for adults, while th e dietary subpopulation youths aged 13 to 19 was chosen to match with the youth gol fer 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 go lfer MOE to yield th e following aggregate

short/intermediate-term MO Es: 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 concl uded that there is reasonable certainty that no harm will result from acute, short/in termediate 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 developm ental neurotoxicity8

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 multigenerati on 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 a nd 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 20.5% of the aPAD for the “Children (1-2 years)” subgroup, and 51.0% of the cPAD for the “All Infants (<1 year) “subg roup. (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 ch ildren 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 and Mexican MRLs for beans and potatoes are similar to the tolerances established for these crops in the USA.