EPA REGISTRATION DIVISION COMPANY NOTICE OF FILING FOR 

PESTICIDE PETITIONS PUBLISHED IN THE FEDERALREGISTER

Isagro, S.p.A.

EPA has received a pesticide petition (3E8162) from Technology Sciences Group 

on behalf of Isagro S.p.A., 1150 18th Street NW, Suite 1000, Washington, DC 20036

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

1. by establishing a tolerance for residues of

Benalaxyl-M import tolerances for Grape, 1.1 ppm; Grape, juice 1.1 ppm; Grape, wine 

1.1 ppm; Grape, raisin 2.2 ppm; Tomato 0.25 ppm and Tomato, processed 0.25 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

1. Plant metabolism. In plants and animals, the metabolism of Benalaxyl-M is

adequately understood for the purposes of establishing the proposed tolerances. Based on 

the available metabolism and toxicology data, Benalaxyl-M is proposed to be considered 

as the residue of concern in plant and animal matrices.

2. Analytical method. In plants and animals, the residue of concern, Benalaxyl-M, 

can be determined using Liquid Chromatography (LC) with a Mass Spectrometer (MS) 

detector for the proposed uses:

For imported commodities: grape, raisins, and grape processed commodities at 10 parts 

per billion (ppb), tomato and tomato processed commodities at 10 parts per billion (ppb).

Studies to evaluate FDA multi-residue method testing were performed for plant samples.

3. Magnitude of residues. To support the import tolerance request, the registrant 

has provided 25 grape and 23 tomato Benalaxyl-M residue trials that mimic product use 

conditions and rates to support an import tolerance in the United States. In addition, 

residue studies were conducted on processed products (i.e., raisins, grape juice, young 

wine, bottled wine, vegetable (tomato) juice, ketchup, canned tomatoes, pureed tomatoes, 

and tomato paste). The residue demonstrates that imported commodities treated with 

Benalaxyl-M will be below the proposed tolerances.

B. Toxicological Profile

1. Acute toxicity. The acute toxicity studies indicated low toxicity for 
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technical Benalaxyl-M, placing it into Toxicity Category III for oral and dermal toxicity, 

Toxicity Category III for eye irritation, and Toxicity Category IV for skin irritation. 

Dermal sensitization studies found that Benalaxyl-M is a nonsensitizer.

2. Genotoxicty. Benalaxyl-M does not induce gene mutations in bacterial and 

mammalian cells, chromosome aberrations in mammalian cells or deoxyribonucleic acid 

(DNA) damage in bacterial cells in in vitro test systems. Similarly, it does not exhibit a 

clastogenic effect in vivo in the mouse micronucleus test. Therefore, there is no evidence 

to suggest a genotoxic hazard.

3. Reproductive and developmental toxicity. In the rat developmental toxicity 

study with Benalaxyl-M, there was no evidence of teratogenicity or other embryotoxic 

effects at the highest dose levels although maternal toxicity was evident based on 

decreased body weight gain at 250 mg/kg bw/day, the highest dose level tested A2 05 06 

01. In the rabbit study test the maternal NOAEL was 1,000 mg/kg bw/day, the highest 

dose level tested. The developmental NOAEL in the rabbit developmental toxicity study 

was found to be 250 mg/kg bw/day based on developmental delays at 1,000 mg/kg 

bw/day. A two generation reproduction study in the rat found a NOAEL of 55.6 to 81.0 

mg/kg bw/day in males and 70.7 to 116 mg/kg bw/day in females based on decreased 

body weight gain in F1 and F2 pups during lactation at the high dose level. Liver effects 

observed in high dose parents. The effects reported at the dose level were considered to 

be adaptive in nature.

4. Subchronic toxicity. A 90-day rat study found a NOAEL of 80 ppm, equal 

to 6.7 mg/kg bw/day, based on reduced weight gain in females receiving 400 ppm and 

greater A2 05 03 02. A 90-day study in the dog reported a NOAEL of 900 ppm, equal to 

32.0 mg/kg bw/day for males and 35.0 mg/kg bw/day for females, based clinical 

chemistry changes indicative of liver toxicity. A 90-day rat neurotoxicity study reported a 

NOAEL above 10000 ppm (highest tested dose), equal to 784 mg/kg bw/day for males 

and 1139 mg/kg bw/day for females for neurotoxicological effects. An immunotoxicity 

study conducted for 28-days in male rats with diets containing up to 10,000 ppm of 

Benalaxyl-M resulted in no evidence of immunotoxicity (ref. A2 05 10 01). The NOEL 

was equal to 924.6 mg/kg bw/day, the highest dose level tested. A 28-day dermal toxicity 

in the rat is available for Benalaxyl-M. No evidence of adverse effects was observed at 

any dose level (ref. A2 05 03 08). The NOAEL was determined to be 1,000 mg/kg 

bw/day, the highest dose level tested.

5. Chronic toxicity. A one year study was conducted with Benalaxyl-M in the 

dog. Animals were dosed by capsule for 52 weeks. The NOEL was found to be 100 

mg/kg bw/day based upon clinical chemistry changes indicative of liver toxicity at a dose 

level of 300 mg/kg bw/day. Diffuse hypertrophy of the thyroid was seen at this dose 

level in females.

A chronic study in the rat was conducted to assess chronic toxicity and 

oncogenicity (ref. A2 05 05 01). Rats were exposed to Benalaxyl-M for 52 weeks in 

chronic phase of the study and for 104 weeks in the oncogenicity assay. The NOEL for 

chronic toxicity was determined to be 60 ppm (equal to 2.4 mg/kg bw/day for males and 
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3.13 mg/kg bw/day for females) based upon treatment-related effects in liver and thyroid 

gland of both sexes at higher dose levels. There was no evidence of a neoplastic response 

at any dietary concentration in the rat study. 

An oncogenicity study of Benalaxyl-M in the mouse was conducted for 78 weeks. 

An increased incidence of hepatocellular adenomas was observed in males and females of 

the high dose level of 7,000 ppm (equivalent to 962.23 and 1180 mg/kg bw/day in males 

and females respectively). A mode-of-action study conducted subsequent to the mouse 

oncogenicity study indicated that the tumor response in mice was associated with CYP 2b 

induction and a pattern of cell proliferation and enzyme induction similar to 

phenobarbital (ref. A2 05 05 03). The weight of the evidence indicates that Benalaxyl-M

is not likely to be a human carcinogen.

6. Animal metabolism. A metabolism study in rats found the same metabolic 

pathway for the R and S enantiomers. This pathway consists of oxidation of the methyl 

group of the aniline ring to hydroxymethyl and then to carboxylic acid. Hydroxylation 

of the phenyl ring and hydrolysis of the carboxylmethyl group also occurs. 

7. Metabolite toxicology. Two of the soil metabolites have been tested in 

genotoxicity batteries and 90-day studies in rats. The methyl-N-malonyl-N (2,6-xylyl)-

D,L-alaninate metabolite was negative for genotoxicity in the reverse mutation, mouse 

lymphoma, and micronucleus assays. This metabolite was clastogenic only in the 

absence of S9-activation in the chromosomal aberration assay. The NOAELs in a 90-

day study in the rat by dietary incorporation were 922.8 and 1072.6 mg/kg bw/day for 

males and females respectively. The N-malonyl-N-(2,6-xylyl)-D,L-alanine metabolite 

was negative for genotoxicity in the reverse mutation, mouse lymphoma and 

chromosomal aberration assays. The NOAELs in a 90-day rat study by dietary 

incorporation were 819.2 and 978.1 mg/kg bw/day for males and females, respectively.

8. Endocrine disruption. To date there is no evidence to suggest that Benalaxyl-M 

functions in a manner similar to any known hormone, or that it acts as an endocrine 

disrupter.

C. Aggregate Exposure

1. Dietary exposure. Estimates of chronic dietary exposure were calculated using 

Dietary Exposure Evaluation Module Food Commodity Intake Database (DEEM TMFCID TM) software from the US EPA. The database was used to determine chronic 

exposure estimates for the overall U.S. population and 24 population subgroups. 

The resulting dietary exposures were compared to a chronic reference dose (RfD) of 

0.024 mg/kg bw/day. The RfD is based on the NOAEL of 2.4 mg/kg bw/day from the rat 

chronic toxicity study (52 week) and the rat carcinogenicity feeding study (104-week) 

with a 100-fold uncertainty factor to account for intra-species and inter-species 

variations. 

i. Food. Chronic dietary exposure estimates for the overall U.S. population and 24 

population subgroups, including infants and children, are well below the chronic RfD. 
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Estimated daily exposures from tolerance level residues and a 100% CT assumption for 

all crops were 15.8% of the RfD or less for all populations examined. The assumption of 

100% crop treated is a worst case for Benalaxyl-M.

ii. Drinking water. No exposures via drinking water are expected based on the 

proposed import tolerances.

2. Non-dietary exposure. No exposures via residential uses are expected based on 

the proposed import tolerances. There are currently no residential uses for Benalaxyl-M.

D. Cumulative Effects. Consideration of a common mechanism of toxicity is not 

necessary at this time because there is no indication that toxic effects of Benalaxyl-M 

would be cumulative with those of any other chemical compound.

E. Safety Determination

1. U.S. population. Benalaxyl-M is a new active ingredient that is proposed 

for import tolerances on grape, raisin, and tomato. It is the R isomer of the racemic 

mixture, Benalaxyl, which was evaluated by the Joint Meeting on Pesticide Residues 

(JMPR) in 2009. JMPR proposed Maximum Residue Limits for Benalaxyl on tomatoes, 

grapes, melons, onions, potatoes, lettuce and meat milk and offal. Using the chronic 

exposure assumptions for the proposed tolerances and the proposed RfD described above, 

the dietary exposure for the U.S. population was calculated to be 3.4% of the RfD. The 

maximum dietary exposure to a population subgroup in the United States is calculated to 

be 15.8% for children 1-2 years old if it is assumed that 100% of the each of the crops are 

treated and all grapes and tomatoes consumed in the U.S. are imported. No residential or 

drinking water exposure is anticipated. Therefore, taking into account the proposed uses, 

it can be concluded with reasonable certainty that residues of Benalaxyl-M in food and 

drinking water will not result in unacceptable levels of human health risk.

2. Infants and children. FFDCA section 407 provides that EPA shall apply an 

additional safety factor for infants and children to account for prenatal and postnatal 

toxicity and the lack of completeness of the database. Only when there is no indication of 

increased sensitivity of infants and children and when the data base is complete, the extra 

safety factor may be removed by the EPA. In the case of Benalaxyl-M, the toxicology 

database is complete and there is no indication of increased sensitivity in the 

developmental and multi-generation reproductive toxicity studies. Therefore, it can be 

concluded that there is no need for an additional safety factor and a safety factor of 100

can be used for the assessment. Chronic and multigenerational studies on mammals, 

aquatic organisms and wildlife did not reveal any immune or endocrine effects. Any 

endocrine related effects would have been detected in this array of required tests.

F. International Tolerances. A Codex Alimentarium Commission Maximum Residue 

Level (MRL) has not been set for Benalaxyl-M.