EPA Registration Division contact: Andrew Ertman; 703 - 308 - 9367 Interregional Research Project No. 4 PP No. 2E8119 EPA has received a pesticide petition (2E8119) from Interregional Research Project Number 4 (IR - 4) , Princeton, NJ 08540 proposing, pur suant 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. 476 by establishing a tolerance for residues of Triflumizole [ 1 - [1 - ((4 - chloro - 2 - (trifluoromethyl) phenyl)imino) - 2propoxyethyl] - 1H - imid azole] ] in or on the raw agricultural commodity tomato at 1.5 parts per million (ppm); F ruit , pome, group 11 - 10 at 0.5 ppm ; F ruit , small, vine climbing , except fuzzy kiwi fruit, subgroup 13 - 07F at 2.5 ppm; and Berry, l ow growing , subgroup , 13 - 07G at 2.0 ppm . EPA has determined that the petition contains data or information regarding the elements set forth in section 408 (d)(2) of the FDDCA; however, EPA has not fully evaluated the sufficiency of the submitted data at this time or whether the data supports g ranting of the petition. Additional data may be needed before EPA rules on the petition. A. Residue Chemistry 1. Plant metabolism. In crops, the metabolism of [14C]

- Phenyl] triflumizole was investigated in cucumber, pears, grapes and apples. The major metabolites were: N

- (4chloro

- 2trifluoromethylphenyl)

- n - propoxyacetamidine (FM

- 6

- 1), N

- (4

- chloro

- 2

- trifluoromethylphenyl)

- n - propoxyacetanilide (FD

- 1

- 1) and the free or conjugated products of N

- (4

- chloro

- 2

- trifluoromethylphenyl)

- hydroxyacetamidine (the 0

- dealkylation product of FM

- 6

- 1), N

- (4

- chloro

- 2

- trifluoromethylphenyl)

- hydroxyacetanilide (FD

- 2

- 1) and the triflumizole aniline (FA

- 1

- 1). 2. Animal metabolism. In livestock, oral administration of [ 14 C]

- triflumizole was conducted twice daily for 5 days a t 14 mg/kg/day (equivalent to 280 ppm in feed for the cow) and 144 mg/kg/day (equivalent to 50 ppm in feed for chicken). On the sixth day the animals were sacrificed and meat, milk and egg samples were analyzed. Cow’s liver, kidney, fat and muscle contain ed 11.26, 3.36, 0.66, and 0.30 ppm of [ 14 C]

- triflumizole equivalent, respectively. Residues in milk reached a plateau of about 0.7 ppm after 24 hours. Chicken liver, kidney, fat, and muscle contained 1.10, 0.74, 0.17 and 0.05 ppm of [ 14 C]

- triflumizole equi valent, respectively. Residues in eggs reach a maximum of 0.30 ppm level. Metabolites identified in these tissues were mainly triflumizole aniline (FA

- 1

- 1) and the sulfate/glucuronide conjugates of the 6

- hydroxy derivative of triflumizole aniline (FA

- 1

- 5) . The latter was the major metabolite in milk. Unchanged triflumizole was present in trace amounts in the whole egg. 3. Analytical method. The analytical method is suitable for analyzing crops for residues of triflumizole and its aniline containing meta bolites at the proposed tolerance levels. The analytical method has been independently validated. Residue levels of triflumizole are converted to FA

- 1

- 1 by acidic and alkaline reflux, followed by distillation. Residues are then extracted and subjected to S PE purification. Detection and quantitation are conducted by gas chromatograph equipped with nitrogen phosphorus detector, electron capture detector or mass spectrometry detection. The limit of quantitation of the method has been determined in the range of 0.01 ppm to 0.05 ppm for the combined residues of triflumizole and FA

- 1

- 1. The enforcement methodology has been submitted to the Food & Drug Administration for publication in the PesticideAnalytical Manual, Vol. II (PAM II). 4 . Magnitude of residues. Magnitude of residue data were collected from four greenhouse trials located in the EPA Regions 2, 3, 6 and 9 to support the minor use of triflumizole on greenhouse cucumber for the control of powdery mildew. Foliar directed applications were used at all g reenhouse sites at a targeted 0.25 lb a.i./A. Foliar directed applications for all trials were made at 6 to 8 day intervals. The last application was on the day of harvest (0 day PHI) at all trials. A decline study was conducted at one location, with the l ast application 0, 3, 7, 12 and 15 days before harvest (0

- 15 day PHI). The results from these trials show that the maximum residue of triflumizole (total) in greenhouse cucumber at 0 day PHI was 0.27 ppm. Residues of triflumizole (total) in the decline tr ial generally declined from the 0 day sampling (maximum residue of 0.10 ppm) to the 15 day sampling (maximum residue of <0.10 ppm). Residues detected in all control samples were analyzed and found to be <0.10 ppm in triflumizole (total). Magnitude of resi due data have been collected from four greenhouse trials located in NAFTA Regions to support the minor use of triflumizole on greenhouse tomato for the control of powdery mildew. At the three of the trials, four foliar directed applications of the test sub stance 6

- 8 days apart were made to the treated plots. At one trial, five applications of the test substance were made 6

- 14 days apart. The application rates ranged from 0.230 to 0.258 lb a . i
. /A per application for a total rate range of 0.921 to 1.284 lb a. i. /A per season. At all the field trials, samples were harvested 0 days after the last application. Additionally, at one trial, duplicate samples were collected 1, 4, 8, 10 and 15 days after the last application. The total residues of triflumizole range d from <0.50 to 0.88 ppm in samples taken at 0 days PHI. There were no residues above the lowest level of method validation of 0.50 ppm for samples taken at 1, 4, 8, 10 or 15 days PHI. The nature of the residues of triflumizole is adequately understood in greenhouse cucumber and greenhouse tomatoes, and an acceptable analytical method is available for enforcement purposes. B. Toxicological Profile 1. Acute toxicity. The database includes the following studies for triflumizole technical: a rat acut e oral study with a LD 50 of 1.42 g/kg; a rabbit acute dermal study with a LD 50 >5 g/kg; a rat acute inhalation study with LC 50 >3.2 mg/l; a rabbit primary ocular irritation study which showed mild irritation; a rabbit primary dermal irritation study which showed no irritation; a guinea pig dermal sensitization study which showed slight dermal sensitization potential. The database also includes the following studies for Procure 480SC/Terraguard 480SC: a rat acute oral study with a LD 50 >2 g/kg; a rabbit acu te dermal study with a LD 50 >5 g/kg; a rat acute inhalation study with a LC 50 >3.3 mg/l; a rabbit primary ocular irritation study which showed slight irritation; a rabbit primary dermal irritation study which showed no irritation; a guinea pig dermal sensi tization study which was negative for skin sensitization. 2. Genotoxicty. Triflumizole was negative in all genotoxicity assays including: Ames assay in S. typhimurium, gene conversion assay in yeast strain D4, REC assay in B. subtilis, unscheduled DNA synthesis (UDS) assay in rat hepatocytes, chromosome aberration assay in Chinese hamster lung (CHL) cells and a mouse micronucleus assay. 3. Reproductive and developmental toxicity. In a developmental toxicity study inrats, triflumizole was administer ed by oral gavage to pregnant, female Sprague Dawley rats at dosage levels of 0, 10, 35 or 120 mg/kg/day. Maternal toxicity, as evidenced by a substantial reduction in body weight gain, was seen at 35 and 120 mg/kg/day. At these dosage levels there was a d ecrease in fetal viability in the form of late resorptions. There were no teratogenic effects. The no observed effect level (NOEL) for maternal and developmental toxicity was 10 mg/kg/day. In another study Triflumizole was administered by oral gavage to p regnant, female New Zealand White rabbits at dosage levels of 0, 5, 25, or 50 mg/kg/day. Maternal toxicity was observed at a dose level of 50 mg/kg/day as demonstrated by a reduction in body weight gain. There were no developmental or teratogenic effects. The NOEL for maternal toxicity was 25/mg/kg/day and the NOEL for developmental toxicity was greater than 50 mg/kg/day. The reproductive toxicity of triflumizole was evaluated in rat reproduction study, conducted on three generations, at dietary concentrat ions of 0, 30, 70 and 170 ppm. Fertility was not affected by treatment. There was an increase in placental weight in the Flb, F2b and F3b litters and a statistically significant increase in gestation length in the high dose group at the F1a interval and at all three dose levels at the F3a interval. Critical review of the gestation length data demonstrated that the NOEL for reproductive effects in this study is 70 ppm (MRID # 47136402), which was the original conclusion of the study director at Life Science Research. This is based on the increase in gestation length seen at 170 ppm in two of six matings. In addition, the NOEL for systemic parental toxicity was greater than 170 ppm and the NOEL for developmental toxicity was 70 ppm (3.5 mg/kg/day) based upon i ncreased incidences of hydroureter and space between the body wall and organs. 4. Subchronic toxicity. To assess sub

- acute dermal toxicity, triflumizole was applied to the backs of male and female Sprague Dawley rats for 6 hours per day for three weeks at dose levels of 10, 100 and 1000 mg/kg/day. Female rats exposed to 1000 mg/kg/day exhibited mild fatty vacuolation in the liver, which was within the range of normal biological variation. Therefore, the no

- observed effect level (NOEL) for subchronic d ermal toxicity in rats was considered to be greater than 1000 mg/kg/day. In a subchronic feeding study in rats 80 male and 80 female SD rats were administered triflumizole at dose levels of 20,200, and 2000 ppm for 13

- weeks. No remarkable clinical signs we re seen. Histopathological findings indicated accumulation of fat droplets in the peripheral zone of the liver at dose levels of 2000 ppm. From this data, it is concluded that the target organ is the liver and the NOEL is considered to be 200 ppm. In anoth er subchronic study in mice, 80 male and 80 female ICR mice were given diets containing 0, 20, 200, and 2000 ppm of triflumizole for 13

- weeks. No marked clinical signs were seen in the dosed groups except for growth retardation in males of 20 and 2000 ppm groups. A significant increase in liver weight was found in both sexes of the 2000 ppm groups. Histopathological findings showed that the dose

- related change was confined to the males in the 2000 ppm group. The lesion consisted of swollen liver cells in th e central zone. The NOEL was determined to be 200 ppm. Four

- week cumulative toxicity study was conducted in rats and mice at dose levels of 0, 20, 200, 2000 ppm. No marked clinical signs of toxicity were seen. Liver weights and body weight ratios were elev ated in both sexes at 2000 ppm dose levels. Histopathological findings were confined to hepatic lesions in males of the 2000 ppm group, with swollen cells in the central zone. The target organ was liver and the NOEL was determined to be 200 ppm. Neu rotoxicity testing battery included acute and subchronic neurotoxicity studies. In theacute neurotoxicity study male and female rats were administered a single dose of triflumizole by oral gavage at levels of 0, 25, 100, 200 (female) and 400 (male) mg/k g and observed for 15 days. Decreased motor activity was noted 3 to 6 hours post exposure in male and female rats at 100 mg/kg and above. Functional Observational Battery (FOB) effects included a decrease rearing counts in females, decreases in forelimb an d hind limb grip strength in males and forelimb grip strength in females at 100 mg/kg and above. The NOAEL was considered to be 25 mg/kg based on clinical findings and functional observational battery evaluations at 100 mg/kg. In a 90 day neurotoxicity Stu dy male and female rats were administered triflumizole in the diet at dose levels of 70, 700 or 2000 ppm (4.1, 40.9, 117.3 mg/kg/day male and 4.9, 47.8, and 133.3 mg/kg/day females) for a period of 90 days. The NOAEL for general toxicity was 70 ppm based o n body weight (significantly higher in males at 700 ppm and significantly lower in females at 2000 ppm), food consumption (lower in females at 2000 ppm), hematology (values were slightly lower in males and females at 2000 ppm and at week 4 in the 700 ppm d ose group) and pathology evaluations (higher in mean and relative liver weights in males and females at 700 and 2000 ppm, hepatocellular hypertrophy in males and females at 700 and 2000 ppm). The NOAEL for neurotoxicity was 2000 ppm. An immunotoxicity stud y was conducted to determine if triflumizole was immunotoxic by measuring antibody production to sheep red blood cells (SRBC) and natural killer (NK) cell activity in BALB/c female mice treated for 28

- days with 0, 20, 200, and 2000 ppm triflumizole. There were no effects of triflumizole, at any dose on thymus weight, spleen weight, or NK cytolytic activity, as a measure of innate immunotoxicity. Triflumizole at 20 and 200 ppm had no effect on terminal body weight or SRBC IgM production. Treatment with 2000 ppm triflumizole caused a small reduction in terminal body weight, and reduced anti

- SRBC IgM production (0.55x of vehicle). In conclusion, the triflumizole NOAEL in this study was 200 ppm, and the LEL was 2000 ppm. 5. Chronic toxicity. In one year dog f eeding study, triflumizole was fed to male and female beagle dogs for one year at dietary concentrations of 0, 100, 300 and 1,000 ppm to assess chronic toxicity. At a dosage le v el of 1,000 ppm there was an increase in serum liver enzymes and a decrease in RBC concentration. The NOEL for chronic toxicity in dogs was 300 ppm (7.5 mg/kg/day). In a two

- year rat feeding study, triflumizole was fed to male and female Sprague Dawley rats for two years at dietary concentrations of 0, 100, 400 and 1,600 ppm to as sess chronic toxicity. At the high dose level of 1600 ppm there was a substantial reduction in body weight gain in males and females. At the mid and high dose levels of 400 and 1600 ppm there was an increase in liver weight. Fatty vacuolization of the live r was seen at all dose levels and hepatocytic hypertrophy was seen in high and mid

- dose males and females. Female rats given 400 or 1,600 ppm were confined to hepatocytic fatty vacuolation and hypertrophy in females. These changes were less severe than tho se seen in rats given 400 or 1,600 ppm and were considered by the laboratory to be indicative of adaptive metabolic change. No evidence of oncogenic effects was demonstrated in rats. The dietary level of 100 ppm (5 mg/kg/day) is considered to be a NOEL. In two

- year mouse oncogenicity study, triflumizole was fed to male and female B6C3F1 mice for two years at dietary concentrations of 100, 400 and 1,600 ppm. At a dosage level of 1,600 ppm there was a substantial effect on body weight gain in males and a n increase in liver weight in males and females. There was no evidence of oncogenic effects in mice as a result of triflumizole administration. 6. Animal metabolism. Triflumizole, [14C

- Phenyl] 1

- (1

- ((4

- chloro

- 2

-trifluoromethylphenyl)imino)

- 2

- propoxyethy l)

- 1H

- imidazole, was found to be rapidly absorbed and excreted in rats. Two days after oral dosing, 78% was found to be excreted in the urine and 20% in the feces. No sex difference was noted. It appears that the loss of the imidazole ring was the basic st ep in the metabolic pathway of the fungicide in mammals. The elimination of the imidazole ring yielded initially N

- (4

- chloro

- 2

- trifluoromethylphenyl)

- n - propoxyacetamidine (FM

- 6

- 1), and N

- (4

- chloro

2trifluoromethyphenyl

- n - propoxyacetanilide (FD

- 1

- 1). Othe r hydroxylated metabolites identified (free, or as sulfate/glucuronide conjugates) included, among others, N

- (4

- chloro

- 2

- trifluoromethylphenyl)

- hydroxyacetamidine (FM

- 8

- 1); 4

- chloro

- 2

- trifluoromethyl

- hydroxyacetanilide (FD

- 2

- 1); and 4

- chloro

- 2

- trifluorome thyl

- 6

- hydroxyaniline (FA

- 1

- 5). 7. Metabolite toxicology . Both plant and animals produce the same metabolites that were identified in the metabolism studies; therefore, the toxicity of the metabolites has essentially been evaluated in the rat toxicolo gy studies. 8. Endocrine disruption . Based on available studies there are no endocrine effects of concern. C. Aggregate Exposure 1. Dietary exposure. Based on dietary, drinking water and non

- occupational exposure assessments, there is reasonable certainty of no harm to the US population, any population subgroup, or infants and children from acute or chronic exposure to triflumizole. i. Food
. Tolerance level residues were used for the proposed commodities in the acute and chronic assessment. Fo r the chronic assessment, ARs calculated as the mean value from field trial data were used, and the crops were assumed to be 100% treated for the new uses. For existing registered crops, percent crop treated were used, as listed in the June 3, 2009 Federal Register (Vol. 74, No. 105, 26536

- 26543). Chronic dietary exposures were estimated utilizing Dietary Exposure Evaluation Model software with Food Commodity Intake Database (DEEM

- FCID), version 3.12, which incorporates NHANES 2

- day food consumption data f or 2003

– 2008. The updated version also uses food translations based on the EPA/USDA FCID recipe set as of March 2011. The revised cPAD of 0.0117 mg/kg/day was used in this assessment. The total chronic dietary exposures are estimated to be 67.2% of the cPAD for the general US population, 11.8% for all infants, 66.8% for children aged 1

- 2 years old, and 74.3% for children aged 3

- 5 years old. Chronic dietary exposures below 100% of the cPAD are reasonably anticipated to result in no harm to exposed popula tions. Acute exposures were calculated using DEEM

- FCID version 3.15. The acute population adjusted dose (aPAD) for triflumizole is 0.25 mg/kg/day for the general population and 0.1 mg/kg/day for females aged 13

- 49. At the 95 th percentile, which is the app ropriate point of comparison for this conservative Tier 1 acute dietary assessment, the estimated acute exposure range from 4.3% of the aPAD for all infants to 27.7% of the aPAD for children aged 1

- 2 years old, and 28.7% for children aged 3

– 5 years old. Fo r females aged 13

- 49 years old, the 95 th percentile acute dietary exposure is 24.5%. Acute dietary exposures below 100% of the aPAD are reasonably anticipated to result in no harm to the exposed populations. ii. Drinking Water. Tier I screen models FIR ST (First Index Reservoir ScreeningTool) and SCI

- GRO (Screening Concentrations in Ground Water) were used to predict estimates of triflumizole concentrations in surface and groundwater, respectively. Drinking water levels of comparison (DWLOCs) were estim ated and compared to drinking water estimated concentrations (DWECs) using EPA standard operating procedures. DWECs, obtained from the March 17, 2009, EPA’s acute exposures from the use on cherries, was estimated to reach a maximum of 37.4 ug/L. The chroni c drinking water concentration was not expected to reach 12.0 ug/L . The chronic and acute DWECs are below the chronic and acute DWLOCs, respectively, for all population groups and exposure durations. 2. Non - dietary exposure . There are no registered or proposed uses for application by homeowners. For the proposed residential uses, only professional handlers would apply triflumizole to woody ornamentals (trees, shrubs, vines). Triflumizole is not used or proposed for use on lawns or turf. Therefore, resid ential exposure is not likely. D. Cumulative Effects The potential for cumulative effects of triflumizole, an imidazole, and other substances that have a common mechanism of toxicity was considered. The mammalian toxicity of triflumizole is well defin ed. No reliable information exists to indicate that toxic effects produced by triflumizole would be cumulative with those of any other chemical compounds. Therefore, consideration of a common mechanism of to xicity with other compounds is not appropriate. T hus, only the potential risks of triflumizole are considered in the aggregate exposure assessment. E. Safety Determination U.S. population. i . Short

- Term Risk. The acute population adjusted dose (aPAD) for triflumizole is 0.25 mg/kg/day for the ge neral population and 0.1 mg/kg/day for females aged 13

- 49. Acute dietary exposure to children (1

- 2 yrs) was 0.06922 mg/kg/day and was 27.7% of the aPAD. Acute dietary exposure to females 13

- 49 yrs was 0.06115 mg/kg/day, and was 24.46% of the aPAD. Theref ore, acute aggregate exposure from triflumizole is not expected to exceed the aPAD.
ii. Chronic Risk
. Chronic Risk. In the August 23, 2006, Federal Register Notice, EPA used a cPAD of 0.015 mg/kg/day. Since that time, Chemtura has resubmitted documentati on supporting a revised cPAD for Triflumizole of 0.035 mg/kg/day (MRID# 47136402). The chronic Population Adjusted Dose (cPAD) of 0.035 mg/kg bw/day was based on the NOEL of 3.5 mg/kg/day from the rat reproduction study, a 100x uncertainty factor, and a 1x FQPA safety factor. However, in the 2009 HED evaluation (EPA

- HQ

- OPP

- 2007

- 0312

- 0008), EPA reset the FQPA safety factor to 3x, due to the lack of a NOAEL in the critical study. Therefore, the cPAD of 0.0117 mg/kg/day was used in this assessment. The chronic dietary exposure to the general U.S. population was 0.007865 mg/kg/day and was 67.2% of the cPAD. The chronic dietary exposure to children (1

- 2 yrs) was 0.007812 mg/kg/day and 66.8% of the cPAD. The chronic DWLOC was calculated to be 160 ppb, and the mode l

- predicted chronic EECs of 40 ppb for surface water and 3.11 ppb for groundwater are less than this DWLOC. Therefore, chronic aggregate exposure from triflumizole is not expected to exceed the cPAD.2. Infants and children . Acute dietary exposure to children (1

- 2 yrs) was 0.0692 mg/kg/day and was 27.7% of the aPAD. The chronic dietary exposure to children (3

- 5 yrs) was 0.008693 mg/kg/day and 74.3% of the cPAD. F. International Tolerances No Codex MRLs have been established for the requested u ses. Canada has established an MRL of 0.01 ppm for dried shelled pea and bean, except soybean (Crop Subgroup 6C); cereal grains (Crop Group 15, except rice; dry soybeans and peanuts. Ipconazole is currently registered in Argentina, Australia, Belarus, Boli via, Bulgaria, Canada, China, Czech Republic , France, Hungary, Italy, Japan, Kazakhstan, Mexico, Moldova, Poland, Romania, Russian Federation, Ukraine, United Kingdom, Uruguay and the United States.