Document ID: EPA-HQ-OPP-2015-0717-0002
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2015-11-23T05:00Z

EPA REGISTRATION DIVISION COMPANY NOTICE OF FILING FOR PESTICIDE PETITIONS PUBLISHED IN THE FEDERAL REGISTER  

EPA Registration Division contact: PV Shah, 703-308-1846

Jeneil Biosurfactant Company
[IN-10853]
	EPA has received a pesticide petition (IN-10853) from Jeneil Biosurfactant Company, 400 N. Dekora Woods Blvd. Saukville, WI  53080 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.
	to establish an exemption from the requirement of a tolerance for
	Phenyl ethyl acetate (CAS Reg. No. 103-45-7) when used as an inert solvent applied to growing crops or harvested crops under 40 CFR 180.910 in pesticide formulations.  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 NA remove
	1. Plant metabolism.
	2. Analytical method. NA remove
	3. Magnitude of residues. NA remove
B. Toxicological Profile Phenylethyl acetate, a natural component of a variety of foods, metabolizes readily to innocuous products, as described in Section 7. Therefore, oral toxicity resulting from the use of phenylethyl acetate as an inert ingredient in pesticides is not expected to be significant. To further evaluate the toxicity of phenylethyl acetate, several data sources were utilized:  the safety evaluations for reaffirmation of GRAS status performed by the Expert Panel of the Flavor and Extract Manufacturers Association (FEMA); the Joint FAO/WHO Expert Committee on Food Additives' (JECFA's) Food Additive Series on phenylethyl alcohol, aldehyde, acid and esters and related substances; the Research Institute on Fragrance Materials (RIFM's) toxicological and dermatological assessment of aryl alkyl alcohol simple acid ester derivatives; and U.S. EPA's HPV Challenge Program dossier on phenylethyl alcohol submitted by the Flavor and Fragrance High Production Volume Chemical Consortia/ Aromatic Consortium (FFHPVCC).  
Toxicity data on phenylethyl alcohol and phenyl acetic acid are also considered relevant in this analysis because the metabolism of phenylethyl acetate proceeds rapidly to generate these two compounds. In addition, toxicity of other phenethyl esters and phenoxyethyl alcohol or phenoxyacetic acid is thought to be relevant to that of phenylethyl acetate because 1) they all have a 2-phenethyl or 2-phenoxyethyl carbon skeleton containing a primary oxygenated functional group; 2) they all have similar pharmacokinetic properties and go through nearly identical metabolic pathways, as described above, ultimately leading to the production and excretion of phenylacetic acid or phenoxyacetic acid; and 3) all of these compounds are natural dietary components.
	1. Acute toxicity.  Acute toxicity tests categorize phenylethyl acetate into acute toxicity category III or IV for oral exposure and category IV for dermal and inhalation exposure. Oral LD50 values for phenylethyl acetate in rats range from 3700 mg/kg bw to 5200 mg/kg bw.  The dermal LD50 in rabbits was 6210 mg/kg bw in one study and reported as <10,000 mg/kg bw in another. The inhalation LC50 in rats was >500 mg/m[3].
	2. Genotoxicty. The Research Institute for Fragrance Materials, Inc. (2012) reported results of three Ames assays performed on phenylethyl acetate, two of which were OECD guideline 471 studies. Results of all three showed no significant increase in reverse mutations in Salmonella typhimurium strains TA98, TA100, TA1535, or TA1537 in the presence or absence of metabolic activation at concentrations up to 5000 μg/plate phenylethyl acetate. The genotoxicity of phenylethyl alcohol and six of its derivatives to in vitro and bacterial systems is presented in the following table.
Table 4  -  In Vitro Genotoxicity of Phenylethyl alcohol and its Derivatives (from JECFA 2003)
Compound
Endpoint
Organism/cell type
Maximum concentration
Result             

Phenylethyl alcohol

Reverse mutation

S. typhimurium TA98, TA100, TA1535, TA1537

3 mmol/plate

Negative

Sister chromatid exchange

Human lymphocytes

Not specified

Negative

Phenylacetaldehyde

Reverse mutation

S. typhimurium TA98, TA100, TA104

Not specified
Negative

Mutation

E. coli WP2uvrA/pkM101

Not specified
Negative

Phenylacetic acid

Reverse mutation

S. typhimurium TA98, TA100, TA1535, TA1537, TA1538

1000 mg
Negative

Unscheduled
DNA synthesis

Rat hepatocytes

500 mg

Negative

Mutation

Mouse lymphoma L5178Y Tk+/ -  cells

1500 mg
Negative

Ethyl phenylacetate

Mutation

B. subtilis H17 (rec + ) & M45 (rec  - )

21 mg/disc

Negative

Mutation

B. subtilis H17 (rec + ) and M45 (rec  - )

20 ml/disc

Positive

Reverse mutation

S. typhimurium TA92, TA94, TA98, TA100, TA1535, TA1537

5 mg

Negative

Chromosomal aberration

Chinese hamster fibroblast cells

1 mg/ml

Negative

Mutation

E. coli WP2uvrA (trp - )

200 - 1600
ug/plate

Negative

Isobutyl phenylacetate

Reverse mutation

S. typhimurium TA97, TA102

0 - 0.1 mg/plate   

Negative

Isoamyl phenylacetate

Mutation

B. subtilis H17 (rec + ) and M45 (rec  - )

20 mg/disc

Positive

Mutation

B. subtilis H17 (rec + ) and M45 (rec  - )

20 ml/disc

Negative

Reverse mutation

S. typhimurium TA98, TA100

10 mg/plate

Negative

50 mg/plate

Lethal

para- Tolylacetaldehyde

Reverse mutation

S. typhimurium TA100

0.1 - 1000
ug/plate

Negative

Mutation

E. coli PQ37

Not specified

Negative

In vivo assays of genotoxicity were negative in three chemicals structurally related to phenylethyl acetate. First, ester isoeugenol phenylacetate did not significantly increase the number of micronucleated polychromatic erythrocytes in NMRI mice following intraperitoneal (ip) injections of 1100-2800 mg/kg bw. This compound also did not increase the frequency of sex-linked lethal mutations in Drosophila melanogaster receiving 25 mmol/L for three days. Similarly, there was no significant and biologically-relevant increase in micronucleated polychromatic erythrocytes in mice following ip injections of 625 - 1875 mg 2-phenoxyethyl isobutyrate/kg bw or gavage doses of 500 - 2000 mg/kg bw sodium 2-(4-methoxyphenoxy)propanoate.

	3. Reproductive and developmental toxicity.  Although there were no developmental toxicity data on phenylethyl acetate, there were several studies identified for phenylethyl alcohol and phenylacetic acid, the two principal metabolic products of phenylethyl acetate. Old developmental toxicity screening studies of gavage-administered phenyl ethyl alcohol found teratogenic effects resembling Fetal Alcohol Syndrome. When subsequent developmental and reproductive studies failed to reproduce these findings using dermal or dietary administration routes, a study of the pharmacokinetics of phenyl ethyl alcohol was undertaken. Pharmacokinetic studies of phenylethyl alcohol showed that peak plasma levels of the metabolite phenylacetic acid, which was determined to be the chemical associated with observed toxicity, were significantly higher when administered by gavage compared to other routes. Dermal application garnered less than one-tenth that of gavage with respect to area under the curve and peak plasma concentration. The lowest in vivo concentrations were seen after dietary administration.  Therefore, the results of the dietary and dermal studies presented here are thought to more accurately represent a realistic exposure scenario for phenylethyl alcohol or acetate compared to the gavage results.
Phenyl ethyl alcohol was fed in a microencapsulated form to Sprague-Dawley rats during GD 6 - 15 at doses of 0, 50,150 or 500 mg/kg bw/day. The actual intake was calculated to be 83, 266, and 799 mg/kg bw/day for the three dose groups. Animals were sacrificed on GD20 and litter values were assessed and fetuses evaluated for structural malformations or anomalies. The effect on dams at the high dose was decreased food consumption, resulting in slight weight loss during the first 2 days of treatment. Malformation incidence was not dose-dependent and did not differ significantly in quantity or type between dose groups and control. An increased incidence of incomplete calcification was seen in fetuses of high dose and this was considered to be a possible consequence of the impairment of maternal weight gain. No other evidence of embryotoxicity or developmental toxicity was reported. The NOELs from this study for both maternal and developmental effects were 266 mg/kg/day, and the LOELs were 799 mg/kg/day. 
Sprague-Dawley rats received topical doses of 140, 440 or 1400 mg/kg bw/day phenyl ethyl alcohol on GDs 6-15. Dams receiving 1400 mg/kg bw/day exhibited clear signs of systemic toxicity including death or sacrifice of 3/35 rats, irritability, hunched posture, walking on toes, piloerection, a brown "deposit" in the area of contact, and suppression of mean food intake and growth rate. Animals recovered after cessation of dosing. One non-pregnant female rat at the middle dose displayed similar clinical signs of toxicity as high dose rats, but no other adverse effects were noted in pregnant or non-pregnant females at this dose. The NOAEL for maternal effects was determined to be 440 mg/kg/day.
At the high dose there was an increase in resorptions, a decrease in mean litter size and fetal weight, a wide range of soft tissue and skeletal changes, and incomplete ossification. At the middle and low doses, there were clear dose-related increases in certain visceral abnormalities that were: 1) not seen in controls, 2) not historically common in that strain of rat, and 3) also increased significantly in the high dose group. The pattern of responses suggested a relationship with dose, but was not strictly linear. In the skeletal exam, there was clear evidence of a dosage-related trend in the increased incidence of litters and fetuses with cervical rib(s) and defects of thoracic vertebrae beginning at the lowest dose. The incidences of moderate, commonly observed abnormalities in ossification were clearly dose-related, though not significant at the low dose. Litter parameters were not affected. Authors concluded that overall, there was conclusive evidence of an effect on development at the high dose and clear evidence of an effect at the mid-dose. The slight differences from control values observed at the lowest dose were not statistically significant. The NOAEL for developmental effects of phenyl ethyl alcohol by the dermal exposure route is140 mg/kg bw/day and the LOAEL is 440 mg/kg/day.
A follow-up dermal study looked specifically at cervical rib bud and thoracic vertebrae effects of phenyl ethyl alcohol.  Pregnant rats were treated topically with 70, 140, 280, 430 or 700 mg/kg bw/day on GD 6-16. The incidence of cervical rib buds at the highest dose was statistically significantly higher than controls; there were no significant vertebrae effects. In all dose groups, there was significant, dose-related skin irritation in dams. The LOAEL for maternal effects was 70 mg/kg bw/day, the lowest dose. At all doses, fetuses showed seemingly reversible delayed ossification, which was not dose-related in the two lower dose-groups. Conservatively, the NOAEL for developmental effects in this study was 140 mg/kg/day, however effects seen at this dose may have been associated with maternal irritation and therefore a true developmental NOAEL cannot be derived from these data. 
In summary, developmental effects were not observed in rats after oral administration of phenylethyl alcohol in the absence of maternally toxic doses. Dermal exposure, on the other hand, produced developmental toxicity at 440 mg/kg/day. This difference in toxicity based on route of exposure is consistent with previous pharmacokinetic data on phenyl ethyl alcohol in which dermal penetration and peak plasma concentrations in rats were shown to be higher compared with dietary administration.
Repeat oral doses of phenethyl phenylacetate or phenyl ethyl alcohol, did not cause an adverse response in reproductive organs, even in animals receiving the highest dose levels (500 mg/kg/day phenethyl phenylacetate and 120 mg/kg/day phenyl ethyl alcohol). A dermal repeat-dose study indicated that a dose of 2000 mg/kg bw/day phenyl ethyl alcohol resulted in an increase in relative gonad weight in male rats. These results suggest that these related chemicals are not reproductive toxins, except at very high doses. 
Female Sprague-Dawley rats (10/group) received gavage doses of phenylacetic acid at 250, 500, and 1000 mg/kg bw/day gavage in either a 1% methylcellulose or corn oil vehicle. Dosing lasted from one week prior to a cohabitation period through gestation, parturition, and a 4-day postpartum period, for a total dosing duration of 39 days. Phenylacetic acid at the lowest dose (250 mg/kg/day) produced significant clinical signs of toxicity in dams (excess salivation, ataxia, urine staining), reduced body weight and body weight gain throughout premating, gestation, and lactation, and reduced feed consumption. These effects were reported at higher doses too, along with increased mortality. The NOAEL for systemic toxicity in dams was less than 250 mg/kg bw/day. Fertility parameters and gestation length in treated dams were not statistically different from those of control rats. Offspring of dams in the high dose group showed decreased viability during postpartum days 1-4 and decreased body weights postpartum compared with control animals. However, because dams at this high dose were clearly affected by the test substance, it is not possible to differentiate this effect on reproduction from the maternal effects. Furthermore, as previously stated, pharmacokinetic studies showed that peak plasma levels of phenylacetic acid were significantly higher when administered by gavage compared to other routes. Therefore results observed in the dams in this study are probably not representative of real world exposures. 
	4. Subchronic toxicity. Like phenylethyl acetate, phenethyl phenylacetate will produce the metabolites phenylethyl alcohol and phenylacetic acid.  Toxicity of these two compounds to mammalian systems, therefore, will be very similar. The oral toxicity of phenethyl phenylacetate was assessed by the U.S. Food and Drug Administration (FDA) in Osborne-Mendel rats (10/sex/dose). Animals received diets containing 0, 1,000, 2,500 or 10,000 ppm phenethyl phenylacetate, for an average daily intake of 0, 50, 250 or 500 mg/kg bw, for 17 weeks. At the end of that period, the liver, kidneys, spleen, heart, and testes were weighed and histological evaluation was carried out on these organs as well as the remaining abdominal and thoracic viscera, bone, bone marrow, and muscle. Detailed microscopic examinations were performed on a subset of the rats in the high dose and control groups. There was no effect of the test compound on body weight, food intake, hematological endpoints, or organ weights, and nothing observed in the gross examination of tissues, or histological examination at any dose. The NOAEL therefore was 500 mg/kg bw/day phenethyl phenylacetate, the highest dose.
A 90-day dermal toxicity study was carried out on Sprague-Dawley rats (15/sex/group) phenyl ethyl alcohol at daily doses of 0, 250, 500, 1,000 or 2,000 mg/kg bw. At the two highest dose groups test animals exhibited a statistically significant lower growth rate than controls. There was also a statistically significant decrease in hemoglobin and white blood cell count in males and increases in relative brain, kidney and gonad weights at the high dose only. No other significant effects were noted in the clinical examination or hematology and urine analysis. No findings were reported upon histopathological examination of any organ or tissue, including: adrenals, brain, heart, kidneys, liver, lung, mesenteric lymph node, pituitary, sternum, spinal cord, testes with epididymides, ovaries, spleen, urinary bladder and nerve. The study author reported a NOAEL of 500 mg/kg bw/day, corresponding to an internal dose of 350 mg/kg bw/day.
	5. Chronic toxicity. The oral toxicity of phenylethyl alcohol was assessed in rats as one component in a mixture. Twenty Wistar albino rats per sex received a mixture of compounds dissolved in their drinking water for 56 weeks. The mixture comprised phenylethyl alcohol at 120 mg/kg bw (0.12%), ethyl alcohol at 6000 mg/kg bw (6%), ethyl acetate at 4 mg/kg bw (0.004%), isoamyl alcohol at 120 mg/kg bw (0.12%), isobutyl alcohol at 200 mg/kg bw (0.2%) and acetic acid at 200 mg/kg bw (0.2%). A control group received tap water only. Body weights were monitored throughout and activities of alcohol dehydrogenase, alanine and aspartate aminotransferases, and protein content of liver were determined at 2 - 4-week intervals.  There were no differences between control and treatment groups with respect to weight, hematology, or histology of liver, kidneys, heart, spleen and lungs. Hence, there was no significant effect of chronic exposure to 120 mg/kg/day of phenylethyl alcohol on rats.
	6. Animal metabolism. Studies of the metabolism of aromatic esters, including phenylethyl acetate, in simulated gastric and pancreatic juices indicate hydrolysis is rapid and precedes absorption.  The hydrolysis products are absorbed very quickly after entering the gastrointestinal system. 
Phenylethyl acetate forms phenylethyl alcohol and acetic acid via reactions of the carboxylesterases or esterases, which predominate in hepatocytes but are present in most tissues throughout the body, including small intestine, colon, kidney, trachea and lungs. Acetic acid will be conjugated and excreted via urine or will undergo β-oxidation in the fatty acid metabolic pathway. Phenylethyl alcohol is oxidized via alcohol dehydrogenase to phenylacetaldehyde and then to phenylacetic acid via aldehyde dehydrogenases (ALDH). The Km and Vmax values of human mitochondrial and cytosolic ALDH isoenzymes, as well as the ubiquity of these enzymes, indicate rapid conversion to phenylacetic acid. Phenylacetic acid is an endogenous component of human urine formed during the breakdown of phenylalanine by intestinal bacteria or by oxidative deamination of endogenous phenethylamine. Conjugation of phenylacetic acid is dose-dependent and species-specific. The major conjugates are glucuronic acid, glycine, taurine or glutamine; some is eliminated as the free acid. Results of studies using radio labeled phenylacetic acid concluded that this compound is rapidly absorbed and quantitatively excreted within 24 hours. 
	7. Metabolite toxicology. NA Remove
	8. Endocrine disruption. Toxicity data related to endocrine disruption were not identified in the phenylethyl acetate database. As the scientific knowledge develops, screening of additional compounds may be added to the Endocrine Disruptor Screening Program (EDSP). When additional screening and/or testing is conducted, phenylethyl acetate may be the focus of screening and/or testing to better characterize effects related to endocrine disruption.
C. Aggregate Exposure
	1. Dietary exposure. The estimated dietary exposure to phenylethyl acetate was determined using methods to estimate chronic dietary exposure for a generic inert ingredient. This assessment considers drinking water and crop-specific residues from pre-harvest applications of agricultural insecticides, herbicides and fungicides, assuming the highest established tolerance level residue for each commodity.  The assessment assumes that the inert ingredient is used on all crops, and that 100% of all crops are treated with the inert.  The inert ingredient is assumed to be present in all commodities treated with 57 of the most significant active ingredients at the maximum tolerance level as identified by the U.S. EPA for the default assessment.  Chronic dietary exposure estimates were derived for the general US population and sub-groups of the population using the Dietary Exposure Evaluation Model, DEEM(TM).  The estimated chronic exposure for the total US population is 0.189 mg/kg/day, 3.79% of the chronic Population Adjusted Dose (cPAD).  Children age 1 to 2 years old have the highest estimated exposure at 0.706 mg/kg/day, or 14.13% of the cPAD.
	i. Food. Dietary exposures of concern are not anticipated for phenylethyl acetate due to its ready biodegradation in the environment and low general toxicity. Residues will be minimal and, particularly in relation to background dietary consumption, not an important dietary source.
	ii. Drinking water. Phenylethyl acetate exposure via drinking water from use as an inert ingredient is not expected to be significant. The phenylethyl acetate that enters the water will readily biodegrade.  Exposure via the drinking water was estimated in the DEEM dietary exposure assessment assuming a concentration of 100 ppb.
	2. Non-dietary exposure. Residential exposures of concern are not anticipated for phenylethyl acetate due to its ready biodegradation in the environment and low general toxicity. 
D. Cumulative Effects
	Section 408(b)(2)(D) (9v) of the FFDCA requires that, when considering whether to establish, modify, or revoke a tolerance, the Agency consider "available information" concerning the cumulative effects of a particular pesticide's residues and "other substances that have a common mechanism of toxicity." To our knowledge there are no available data or other reliable information that suggests toxic effects produced by phenylethyl acetate would be cumulative with those of any other chemical compounds. EPA has not made a common mechanism of toxicity finding as to phenylethyl acetate and other compounds. Phenylethyl acetate does not produce toxic metabolites in common with other substances of potential concern. For the purpose of the tolerance exemption proposed, it is assumed that phenylethyl acetate does not share a common mechanism of toxicity with other substances. 
E. Safety Determination
	1. U.S. population. There is a reasonable certainty that no harm to humans will result from the use of phenylethyl acetate as an inert ingredient in pesticide products.  The oral Reference Dose (RfD) and chronic Population Adjusted Dose (cPAD) were derived based on the NOAEL of 500 mg/kg/day from the FDA 17-week dietary study of phenylethyl phenylacetate. For the total US population, the estimated chronic dietary exposure from food and drinking water for Phenylethyl acetate, calculated as 50% of all agricultural formulations, is 3.79% of the cPAD, well below any level of potential concern.
	2. Infants and children. There is a reasonable certainty that no harm to infants and children will result from the use of phenylethyl acetate as an inert ingredient in pesticide products. Phenylethyl acetate is of low toxicity at doses expected from its inert use in pesticide formulations. The structurally similar and metabolically-related compounds phenethyl phenylacetate, phenylethyl alcohol and phenylacetic acid did not produce consistent evidence of developmental or reproductive toxicity following oral or dermal exposure for doses that were not also maternally toxic, nor was there evidence for endocrine disrupting properties. Therefore, there is no concern for increased sensitivity to infants and children to phenylethyl acetate when used as an inert ingredient in pesticide formulations. For this reason, a safety factor analysis has not been implemented to assess risk and the additional tenfold safety factor for the protection of infants and children is also unnecessary. The estimated chronic dietary exposure from food and drinking water was highest (14.13% of cPAD) for children aged 1 to 2 years old.  This estimated exposure to phenylethyl acetate is well below any level of potential concern. 

F. International Tolerances
NA Remove