Page 2 of 32

including benzyl acetate and reconfirmed the group as “GRAS”. Approximately one

- third of the current dietary exposure to benzyl acetate occurs from naturally

- occurring benzyl acetate in a variety of fruit, vegetables and other foods

.

Benzyl acetate exhibits low levels of tox icity via the dermal route of exposure in rabbits and inhalation and oral routes of exposure in rats . It is mildly irritating to the skin and minimally irritating to the eyes in rabbits . It is not a skin sensitizer in guinea pigs

.

In a 13

- week feeding study in the rat, atrophic seminiferous tubules were observed in male rats at 12,500 ppm ( 9 0 0 mg/kg/day )

. The No Observed Adverse Effect Level (NOAEL) was identified

as 6,250 ppm ( 4 6 0 mg/kg/day )

. In mice, following 13 weeks of exposure via the diet, dec reased bodyweight and food consumption w ere observed at all doses. The LOAEL was 3,130 ppm (425 mg/kg/day). The NOAEL was not established.

In a developmental toxicity study, maternal and fetal toxicity were observed at 1000 mg/kg/day. Maternal toxicity was manifested as decreased bodyweight and fetal toxicity was manifested as reduced body weights, increased incidence of dilation of the renal pelvis and skeletal variations. Although qualitative fetal susceptibility is observed, fetal effects occur in t he presence of maternal toxicity and a clear NOAEL of 500 mg/kg/day was established. Therefore, the

established chronic reference dose ( cRfD ( 1.10 mg/kg/day ) ) will be protective of these effects.

The potential for benzyl acetate to be genotoxic was ev aluated in a battery of 10 in vivo mammalian genotoxicity studies. Two tests (a replicative DNA synthesis test at 1600 mg/kg/day and a Comet assay at 1200 and 1600 mg/kg) indicated positive potential to cause genotoxicity at the highest dose (1600 mg/kg). Eight studies were negative for the potential to be genotoxic. A summary of the mammalian genotoxicity studies is attached.

A series of twelve studies of the potential effects of benzyl acetate on several species of bacteria were reported. Two studie s indicated positive effects and one study was reported as yielding equivocal results ( (a ) Rec

- assay with B. subtilis at 20 μ L/disk and ( b ) a microscreen assay with E.

coli at 6.5 μ g/well). Ten studies were negative for the effects of benzyl acetate to indu ce mutagenic changes in bacteria. Based on the weight of the evidence the Agency concludes that benzyl acetate is negative for mutagenic effects. See the attachment for a summary of in vivo

and in vitro studies.

Evidence of neurotoxicity and neuronal de generation was identified in the 13 week NTP studies in rats and mice. Signs of neurotoxicity included tremors and ataxia that were associated with the degeneration of the glial cells in the cerebellum and hippocampus at the doses > 12,500 ppm (2,000/2,98 0 mg/kg/day). Since these effects were induced at doses above than the limit dose (1000 mg/kg/day) and the established cRfD was 1.10 mg/kg/day, these effect s will be protected. Therefore, the concern is low for these effects.

There is evidence that benz yl acetate suppresses immune function in mammalian systems in the

rat occurr ing only at a dose that was lethal and well above the limit dose. In the 13 week feeding study in the rat, a decrease in the cellular components of the bone marrow, thymus and lym phoid follicles were observed at 50,000 ppm (3900 mg/kg/day for males and 4500 mg/kg/day forPage 3 of 32

females), the highest dose tested and well above the limit dose

. The NOAEL for this study was 12,500 ppm (900 mg/kg/day). The potential for immunotoxicity is not of concern because the

effects occur well above the limit dose and the established chronic reference dose (cRfD) will be protective of these effects.

Carcinogenicity studies were conducted by NTP in rats and mice. Benzyl acetate was

administered via th e diet to rats and mice at doses up to 12,000 ppm (510/575 mg/kg/day, male/female). Toxicity was not observed in rats at any dose

. In mice, males and females exhibited reduced body weight throughout the entire study at 345/375 mg/kg/day

.

Prior NTP studi es in rats and mice indicated increased incidence of pancreatic acinar cell adenomas in male rats and the incidences of hepatocellular adenomas and fore

- stomach neoplasms in male and female mice. Because of the confounding effects of the corn oil vehicle

and the controversy over the use of the gavage route of administration, the NTP c onclu ded the corn oil vehicle and the gavage route of administration were the causes of acinar cell adenomas, hepatocellular adenomas and fore

- stomach neoplasms in male and fe male mice.

The No Observed Adverse Effect Level (NOAEL) selected for use in risk assessment is taken

from t he two

- year feeding study conducted by the NTP (1993, MRID 49428712) using B6C3F 1 mice. The NOAEL of 110 mg/kg/day is based on decreased body weig ht seen at 345/375 mg/kg/day and above

. The cRfD was based on this study.

A dermal absorption factor of 35% is identified based on the dermal absorption observed in the rhesus monkey (Bronaugh and Maibac h , 1985, MRID 49428708; Bronaugh et al, 1990, MRID

4 9470803). This value is similar to those seen in the rat.

The request to approve benzyl acetate is for post

- emergent use on growing crops primarily for use in liquid insecticides however some herbicides may be applied to growing crops (harvest

aides etc). Human exposures to benzyl acetate applied as such are expected for those persons occupationally involved in commercial agricultural activities. Exposure is expected by the inhalation and dermal routes. Persons not occupationally exposed due to work activities may be exposed via the oral route of exposure to possible pesticide residues on treated crop commodities or in the case of infants via incidential oral ingestion, and via the dermal and inhalation routes resulting from residential exposures (app licator and post

- application) to pesticide products containing benzyl acetate as an inert ingredient as well as other nonpesticidal consumer products containing benzyl acetate.

Dietary exposure is estimated using the Agency’s Dietary Exposure Estimate Mod el (DEEM). . Estimated exposures do not exceed the Agency’s level of concern

. The estimated dietary exposure to the U.S. Population is 0.189 mg/kg/day which equates to 17.3 % of the c RfD. The estimated dietary exposure to the most highly exposed subpopul ation ( children 1

- 2 years old ) is

0. 691 mg/kg/day which equates to 62.9 % of the c RfD. Dietary Margins of Exposure are:

541 for adults (U.S. Population) and 156 for children 1

- 2 years old ( the most highly exposed subpopulation).Page 4 of 32

A screening level resi dential exposure assessment indicates Margins of Exposure are > 100. The aggregate exposure for adults (food + water + residential applicator and postapplication exposure) results in MOE > 100.

A screening level occupational exposure assessment indicate s all estimated Margins of Exposure

are > 100 except for mixer loaders supporting high acreage aerial operations.

Benzyl acetate is described as moderately toxic to some freshwater organisms. Extensive modeling using the EPISuite TM environmental model system indicates benzyl acetate is readily biodegradable and not likely to bioaccumulate.

Based on the weight of the evidence, the toxicological database for benzyl acetate generally indicates low levels of toxicity, (including mutagenicity and carcinogen icity), dietary exposures not exceeding levels of concern, low occupational and residential exposures and risk and the indication of minimal adverse environmental effects, the Agency approves of the requested exemption from the requirement of a tolerance for benzyl acetate as an inert ingredient (solvent) in pesticide formulations applied to growing crops under 40 CFR 180.920.

II. INTRODUCTION

Technology Sciences Group (TSG) (1150 18 th Street, NW, Suite 1000, Washington, D.C. 20036)

on behalf of the Hun tsman Corporation (8600 Gosling Road, The Woodlands, TX 77381) has

submitted a petition (Submission IN

- 10748; 14 July 2014) for an exemption from the requirement of a tolerance for the compound benzyl acetate (CAS Reg. No. 140

- 11

- 4) applied to

crops pre

- ha rvest

, under 40 CFR § 180.920. The request is for “unlimited use in formulated products applied pre

- harvest.” This assessment is based upon 50% inert ingredient in the final product formulation. As an inert ingredient, benzyl acetate serves as a solvent in pesticide formulations applied pre

- harvest to growing crops.

This is the decision document for benzyl acetate which contains the Agency’s assessment of exposures and risks to human s and the environment as a result of exposures to benzyl acetate.

Benzyl acetate has been used many years as a fragrance in personal care products such as soaps, shampoos, body and facial lotions, fragrance creams and bath products (McGinty et al., 2012 ; MRID 49428701) . Many of these products are “leave

- on” that is

, not required to be rinsed off.

Non

- cosmetic household uses include house

- hold cleaners, detergents and air fresheners McGinty et al. 2012; NLM, HPDB ; MRID 49428701).

Benzyl acetate is approved by the U.S. Food and Drug Administration (FDA) as a direct foo d additive for use as a synthetic flavoring substance under 21 CFR 172.515. The Flavor and Extract Manufacturing Association (FEMA) conducted an updated “generally recognized as safe” (GRAS) review of benzyl derivatives, including benzyl acetate and recon firmed the group

as “GRAS”.

Approximately one

- third of the current dietary exposure to benzyl acetate occurs from naturally

occurring benzyl acetate in a variety of fruit , vegetables and other foods and the other two

- thirdsPage 5 of 32

is from use as a flavoring in gredient (Adams et al., 2005 ; MRID 49428701) . The estimated daily per capita intake of benzyl acetate from the use as a flavoring ingredient by “eaters only” is 14

μ g/kg/day (MRID 49428701).

III. PHYSICAL/CHEMICAL PROPERTIES

Chemical Structure of Benzyl Acetate ( from www.chemicalbook.com )

Table 1.0 Molecular weight (g/mol) 150.176

L og K OW

- 3.35 Boiling Point (°C) 215. 57 Melting Point (°C)

- 0.5 Vapor Pressure mm Hg @ 25°C 1.77 x 10

- 1
Water Solubility 1,605 a
3,100 b

L og K OC 2.059 c

1.944 d
Henry’s Law Constant atm

- m 3 /mole @ 25°C( 2.302 x 10

- 005
Probability of Rapid Biodegradation (BIOWIN) 0.9783 e

0.9988 f
3.0295 days

- weeks g

0.5949 h

0.7562 i
Ready Biodegradability Yes BCF (L/kg wet

- wt) 9.124

Volatilization from Water Half

- Life (hr) 64.71 j

809.1 k
Fugacity Model Compartment/Mass Amount % Air: 2.86

Water: 26.9

Soil: 70

Sediment: 0.173 Fugacity Model Compartment/Half

- Li fe (hr) Air: 40.2

Water: 360

Soil: 720

Sediment: 3.24 x 10 3

a WSKOW modeled result g Biowin3 (Ultimate survey model)

b Experimental database match identified in the model h Biowin5 (MITI linear model)

c MCI method result i Biowin6 (MITI non

- linear m odel)

d K OW method result j Model River

e Biowin 1 (linear model) k Model LakePage 6 of 32

f Biowin 2 (non

- linear model) Physical/Chemical properties taken from MRID 49428701.

IV. TOXICOLOGICAL SUMMARY

Acute Oral Toxicity

– Benzyl acetate exhibits low acute or al toxicity to rats. The oral LD 50 for the rat is 2,490 to 3,690 mg/kg (WHO, 1996 . M RID 49428701 ).

Acute Dermal Toxicity

– The dermal LD 50 in rabbits is > 5,000 mg/kg (McGinty et al., 2012

. M RID 49428701 ).

Acute Inhalation Toxicity

– Rats exposed (w hole body) to 766 μ g/L by the inhalation route for 4 hours exhibited no clinical signs of toxicity on the day of exposure or during the two week

follow

- up observation period. The exposure in the whole

- body chambers w as at the maximum attainable vapor conc entration. The LC 50 was > 766 μ g/L (McGinty et al., 2012. MRID 49428701).

Dermal Irritation

– The potential for benzyl acetate to cause dermal irritation was evaluated in a number of tests including three, 4

- hour exposure studies using the rabbit. A do se of 0.5 ml of benzyl acetate was applied to the dorsal trunk of 3 New Zealand White rabbits and the site was covered by a semi

- occlusive patch. After the 4

- hour exposure, the wrapping was removed from the treatment site and observations for potential de rmal irritation were recorded at 1, 24, 72 and

168 hours. Well

- defined erythema was seen during the observation period that cleared by the end of the study. Desquamation was reported in 1 of 3 rabbits at the end of the study. It was mildly irritating to the skin of rabbits (MRID 49428701)

.

Eye Irritation

– The potential for benzyl acetate to be irritating to the eye was evaluated by the Research Institute for Fragrance Material, Inc. (RIF) in 1994 and summarized by McGinty et al.,

in 2012 (MRID 49428701 ) . A 0.1 ml aliquot of 100% benzyl acetate test article was instilled in one eye of each of three New Zealand White rabbits. Well defined conjunctival irritation was reported that resolved within 48 hours of instillation. Benzyl acetate is considered mi ld ly irritating to the eye

.

Sensitization

– The possibility for skin sensitization was evaluated using guinea pigs (strain not specified). A modified Buehler test was used where 20 albino guinea pigs were exposed to an induction application of 0.3 mL of 100% benzyl acetate on c e weekly for three weeks. Control animals received an induction application of 80 % ethanol/distilled water solution weekly. Challenge patches containing 10% to 30% benzyl acetate in diethyl phthalate and 100% benzyl

acetate were applied to each animal approximately 2 weeks after induction. No sensitization response was produced by benzyl acetate that was greater than the control animals. Benzyl

acetate was determined to be a non

- sensitizer.Page 7 of 32

Subchronic Exposure

Rat

The Nat ional Toxicology Program (NTP 1993 ; MRID 49428712 ) conducted a 13

- week feeding study with the F344/N rat

. Benzyl acetate was administered in the diet at doses of 0, 3,1 3 0, 6,250, 12,500, 25,000 or 50,000 ppm which corresponds to 0, 230, 460, 900, 1750 and 3900 mg/kg/day in 10 males and 0, 240, 480, 930, 1870 or 4500 mg/kg/day in 10 females. Eighteen of the 20 rats in the high dose groups died between weeks 2 and 8 of the study. Also observed at this dose were a decrease in the cellular components of the bone marrow, thymus and lymphoid

follicles. The mean body weight gain and the final mean body weight of 25,000 ppm males were significantly lower ( 10% ) than those of the control gr o up. Feed consumption b y exposed rats, except the 25,000 and 50,000 ppm ma les and 50,000 ppm females was similar to controls. The reduced feed consumption by 25,000 and 50,000 ppm males and 50,000 ppm females m ay have been due to toxicity or decreased palatability. The reduction in body weight coincided with the

decrease in fe ed consumption. Tremors and ataxia occurred only in the 50,000 ppm rats. These finding s were first observed on day 15 in n i ne males and six females and continued until the end of the study. Cholesterol levels at 12,500 ppm and 25,000 ppm and triglycerid e levels in 25,000

ppm in females were lower (14

- 20%) than those in the controls. However these changes are not toxicologically significant. M ild to moderate aspermatogenesis was observed in two 50,000 ppm males and atrophy of seminiferous tubules were o bserved in two 25,000 ppm and one 12,500 ppm male rat. Chemical

- related lesions occurred in the brain, kidney, tongue, and skeletal muscles of the thigh. Necrosis of the brain involving the cerebellum and/or hippocampus, degeneration and regeneration of r enal tubule epithelium and degeneration and sarcolemma nuclear hyperplasia of the tongu e and skeletal muscles occurred in most male and female 50,000

ppm rats. T he no observed adverse effect level (NOAEL) was identified as 6,250 ppm (460 mg/kg/day ) . The lowest observed adverse effect level ( LOAEL ) was 12,500 ppm (900 mg/kg/day ) based on atrophy of seminiferous tubules in males

. ( 2012

, MRID 49428701 ).

Mouse

The NTP (1993. MRID 49428712) conducted a 13

- week study in mice. Ten male and 10 female

B6C 3F 1 mice were fed diets containing 0, 3,130, 6,250, 12,500, 25,000 or 50,000 ppm benzyl acetate equivalent to 0, 425, 1,000, 2,000, 3,700 or 7,900 mg/kg/day for males and 0, 650, 1,280, 2,980, 4,300, or 9,400 mg/kg/day for females. One 50,000 ppm (7,900 m g/kg/day) male mouse died and one 50,000 ppm (9,400 mg/kg/day) female mouse was killed moribund before the end of the study. Mean body weight gains and final mean body weights of all exposed male and

female mice were significantly lower (12

- 33%) than thos e of the controls and the mean body weight gains decreased with increased exposure level. Feed consumption by 3,1 3 0 ppm ( 425/650 mg/kg/day) males and all exposed females was lower (30

- 53%) than that of controls. Tremors occurred only in females and were first observed on day 16 in three females receiving 50,000 ppm, day 94 in one female receiving 25,000 ppm and day 93 in one female receiving 12,500 ppm. The tremors continued until the end of the study. Necrosis of the brain involving the hippocampus occ urred in four 50,000 ppm mice, one male and three females. Hepatocellular

necrosis also occurred in the male with brain lesions. The NOAEL was not established. ThePage 8 of 32

LOAEL was 3,130 ppm (425 mg/kg/day) based on decreased body weights and f ee d consumption.

Developmental Toxicity

– The potential for benzyl acetate to cause developmental toxicity was evaluated by Ishiguro et al., (1993) (summarized in: WHO, 1996) , Adams et al., (2005) and McGinty et al., (2012) (cited in MRID 49428701). Rat dams (strain no t specified) (n = 6 to 7) received doses of 0, 10, 100, 500 or 1000 mg/kg/day of benzyl acetate by gavage for days 6 through 15 of gestation. Data show a decrease in body weight gain of 9.9% on day 6 through day 20 (high dose dams gained 120.8 g compared to control gains of 134.1g). There were no effects of treatment corpora lutea, live fetuses, pre

- or post

- implantation loss, resorptions, sex ratio, or the mean number of fetuses per dam.

Fetal body weights were significantly reduced in the high dose g roup compared to the controls. There was a significant increase in dilation of the renal pelvis in the high dose group offspring in 6/22 litters compared to 2/20 control litters. There was as increased incidence of skeletal variations, including wavy rib s, dumb

- bell shaped vertebrae, lumbar ribs and delayed ossification

in fetuses in the 1000 mg/kg/day group compared to the control group. The skeletal variations reported for the lower dose treatment groups were similar to those in the control group. The study authors concluded that benzyl acetate is not a teratogen and the increased incidences of skeletal and internal variations were due to reduced growth and also reflected the decreased fetal body weight. The maternal and fetal NOAELs for this study we re both 500 mg/kg/day based on a decrease in body weight in dams and reduced body weights, increased incidence of dilation of the renal pelvis and skeletal variations in the offspring at the 1000 mg/kg/day dose.

Genotoxicity /Mutagenicity

The potential for benzyl acetate to be genotoxic was evaluated in a battery of 10 in vivo mammalian genotoxicity studies which are summarized by McGinty et al. (2012). Two tests indicated positive potential to cause genotoxicity at the highest dose (1600 mg / kg). Eight studies were negative for the potential to be genotoxic. A summary of these studies is attached.

A series of twelve studies of the potential effects of benzyl acetate on several species of bacteria were reported by McGinty et al (2012). Two studies indicated positive effects and one study was reported as yielding equivocal results. Ten studies were negative for the effects of benzyl acetate to induce mutagenic changes in bacteria. Based on the weight of the evidence the Agency concludes that benzyl acetate is negative for mutagenic effects. See the attachment for a summary of in vivo and in vitro studies.

Neurotoxicity

Evidence of neurotoxicity and neuronal degeneration were identified in male and female rats in the high dose group (50,000 ppm, 3 900/4500 mg/kg/day for males and females, respectively) in the 13 week NTP study. S igns of neurotoxicity included tremors and ataxia that were associated with the degeneration of the glial cells in the cerebellum and hippocampus. However, these

effects w ere observed at a dose that was lethal and well above the limit dose. Evidence of potential neurotoxicity was also observed in the 13 week study in mice. Tremors were observedPage 9 of 32

in females at > 12,500 ppm (2,000/2,980 mg/kg/day). Since the effects were in duced at doses

above than the limit dose (1000 mg/kg/day) and the established cRfD is 1.10 mg/kg/day, these effect will be protected ; therefore, the concern is low for these effects.

Immunotoxicity

Evidence of potential immunotoxicity was observed only a t a dose that was lethal and well above

the limit dose. In the 13 week feeding study in the rat, a decrease in the cellular components of the bone marrow, thymus and lymphoid follicles were observed at 50,000 ppm (3900 mg/kg/day for males and 4500 mg/kg/d ay for females) , the highest dose tested

. The NOAEL for this study was 12,500 ppm (900 mg/kg/day). The potential for immunotoxicity is not of concern because

the established chronic reference dose (cRfD) will be protective of these effects.

Carcinogenic ity

Rat

The National Toxicology Program (NTP) conducted a 2

- year feeding study of benzyl acetate on rats (NTP, 1993 ; MRID 49428712). G roups of 60 male and female F344/N rats received benzyl

acetate in the diet at concentrations 0, 3,000, 6,000, or 12,00 0 ppm that provided doses of 0, 130, 260 and 510 mg/kg/day for males and 0, 145, 290 and 575 mg/kg/day in females

. Doses were

selected based on the effects seen in the 13

- week study discussed earlier. Survival of exposed

rats was similar to that of the controls. The mean body weights of the 12,000 ppm males and exposed females were approximately 5 % lower than those of the controls throughout most of the study. The feed consumption by 12,000 ppm males was slightly lower than that by the controls.

Diet ary levels of 3,000, 6,000 and 12,000 ppm benzyl acetate were estimated to result in average daily consumption levels of 130, 260, and 510 mg/kg/day for males and 145, 290, and 575 mg/kg/day for females. No biologically significant changes in hematology o r clinical chemistry parameters were found that could be attributed to benzyl acetate. No compound

- related increased incidences of neoplasms or non

- neoplastic lesions occurred in male or female F344/N rats receiving benzyl acetate for 2 years. There is n o evidence of carcinogenic activity in male and female F344/N rats receiving 3,000, 6,000 or 12,000 ppm benzyl acetate in the diet for 2

years. The NOAEL was 12,000 ppm (510/ 575 mg/kg/day , male/female ), the highest dose tested.

Mouse

A two

- year feedin g study was conducted by the NTP (1993 ; MRID 49428712 ) using B6C3F 1 mice that received doses of 0, 330, 1,000, or 3,000 ppm benzyl acetate. That corresponds to 0, 35, 110 or 345 mg/kg/day benzyl acetate for males and 0, 40, 130 and 375 mg/kg/day for

femal es. Survival of all exposed mice except the 3,000 ppm females was similar to that of control groups. Survival of 3,000 ppm females was significantly higher than that of the control group. Throughout the 2

- year study, the mean body weights of 1,000 ppm an d 3,000 ppm males

and females were 9 % to 14 % lower than those of the control animals. No biologically significant changes in hematology or clinical chemistry parameters were observed in mice receiving 330, 1,000 or 3,000 ppm benzyl acetate. No increase in neoplasm incidence in mice could be attributed to benzyl acetate in the feed. Dose

- related increased incidences or severitiesPage 10 of 32

of non

- neoplastic nasal lesions occurred in the most posterior portions of the nasal cavity in all exposed groups. The lesio ns occurred in the majority of the exposed mice and consisted of atrophy and degeneration primarily of the ol f actory epithelium, cy s tic hyperplasia of the nasal submucosal glands, pigmentation of the mucosal epithelium and exudate accumulation. There is

n o evidence of carcinogenic activity of benzyl acetate in male or female B6C3F 1 mice. The

NOAEL was 110/130 mg/kg/day (M/F) . The LOAEL was 345/375 mg/kg/day

, based on decreased body weight

.

The NTP previously studied the toxicology and carcinogenicity of benzyl acetate in F344/N rats

and B6C3F 1 mice using the gavage method of administration and corn oil as a vehicle. Benzyl acetate increased the incidences of pancreatic acinar cell adenomas in male rats and the incidences of hepatocellular adenomas and f ore

- stomach neoplasms in male and female mice. Because of the confounding effects of corn oil on the incidences of pancreatic neoplasm and because of the controversy over the use of the gavage route of administration, the NTP decided

to re

- study benzyl ac etate using the dosed feed route of administration. The conclusions of the 1993 study in rats and mice indicate the corn oil vehicle and the gavage route of administration as the causes of acinar cell adenomas, hepatocellular adenomas and fore

- stomach neo plasms in

male and female mice.

Metabolism/Excretion

– Benzyl acetate is excreted as metabolites primarily in the urine after oral or percutaneous administration. Male F344/N rats given a single dose of 5, 50 or 500 mg/kg

and male B6C3F 1 mice were give n a single oral dose of 10, 100 or 1,000 mg/kg of radioactively labeled benzyl acetate in corn oil excreted 90% of the radioactivity in the urine. None was detected in the adipose tissue, blood, kidney, liver, lung, muscle, skin or stomach. The major

met abolite in the urine was hippuric acid and 95 to 99 % of the excreted dose was in this form. Similar results were observed in male rats given 500 mg/kg and mice given 1,000 mg/kg benzyl acetate in corn oil by gavage once daily, five days/week for two weeks . Neither the size of the

dose or the f

requency of dosing had any effect on the absorption, metabolism or excretion of benzyl acetate.

The percutaneous absorption and disposition of [methylene

14 C] benzyl acetate was studied in F344 rats by applying be nzyl acetate (100, 250, or 500 mg/kg) either neat or as a 50% solution in ethanol over a shaved area on the back of the animal. Twenty four hours after administration 28 to 48% of the dose was recovered from the application site, a similar proportion was absorbed and excreted in the urine and less than 4% remained in the carcass (NTP. 1993; MRID 49428712).

A dermal absorption factor of 35% is identified based on the dermal absorption observed in the rhesus monkey (Bronaugh and Maibac h , 1985, MRID 49428708 ; Bronaugh et al, 1990, MRID 49470803). This value is similar to those seen in the rat.Page 11 of 32

V. TOXICITY ENDPOINT SELECTION

The point of departure f

or benzyl acetate is 110 mg/kg/day from the NTP 2

- year carcin ogeni city study in mice (MRID 49428701) w here throughout the 2

- year study, the mean body weights of

males and females at the LOAEL of 345/375 mg/kg/day (M/F) were 9 % to 14 % lower than

those of the control animals. This endpoint was used for all exposure scenarios. The dermal absorption and inh alation factors were 100%. There was no NOAEL observed in a 90

- day toxicity study in mice based on the effects on body weights seen at all doses (lowest dose tested

was 3,130 ppm ; equal to 425 mg/kg/day) , however, in a carcinogenicity study in mice no effe cts on body weight w ere seen at 110 mg/kg/day. Therefore, 90

- day toxicity study in mice was not

selected.

VI . Special Considerations for Infants and Children

Section 408 of the FFDCA provides that EPA shall apply an additional margin of safety

for in fants and children in the case of threshold effects to account for prenatal and post natal toxicity and the completeness of the database on toxicity and exposure unless EPA determines that a different margin of safety will be safe for infants and children. EPA concludes that the FQPA safety factor could be reduced to 1x for benzyl acetate for the following reasons:

i. The database is considered adequate for FQPA assessment. The following acceptable

studies are available:

Subchronic toxicity (Rat, Mouse) Carcinogenicity (Rat, Mouse) Developmental toxicity (Rat)

ii. Qualit ative fetal susceptibility was observed in the developmental study in rats. Maternal (decrease in bodyweight) and f etal ( reduced body weights, increased incidence of dilation of the

ren al pelvis and skeletal variations) toxicity w ere observed at 1,000 mg/kg/day , the limit dose

. Since fetal toxicity occurs in the presence of maternal toxicity and a clear NOAEL of 500 mg/kg/day was established , the established cRfD ( 1.10 mg/kg/day) will b e protective of these effects. The potential for reproduction toxicity was observed in the 13 weeks dietary study in rats. Atrophy of seminiferous tubules w as observed in males at 12,500 ppm (900 mg/kg/day). However, the c oncern for reproduction toxicit y is low since effects occurred at a high dose and a

clear NOAEL of 6,250 ppm (460 mg/kg/day) was established. Therefore, the established cRfD will be protective of this effect

. In addition, there were no female reproductive parameters

affected in the de velopmental toxicity study in rats.

iii. Neurotoxicity studies were not available for review. Evidence of neurotoxicity and neuronal degeneration w as identified in the 13 week NTP study. Signs of neurotoxicity included

tremors and ataxia that were as sociated with the degeneration of the glial cells in the cerebellum and hippocampus at the high dose (50,000 ppm, 3900/4500 mg/kg/day for males and females, respectively). However, these effects were observed at a dose that was lethal and well above thePage 12 of 32

l imit dose. Evidence of potential neurotoxicity was also observed in the 13 week study in mice.

Tremors were observed in females at > 12,500 ppm (2,000/2,980 mg/kg/day). Since these effects were induced at doses above than the limit dose (1000 mg/kg/day) and the established

cRfD was 1.10 mg/kg/day, these effect will be protected; therefore, the concern is low for these

effects.

iv. Immunotoxicity studies were not available for review. Evidence of potential immunotoxicity was observed only at a dose tha t was lethal and well above the limit dose. In the 13 week feeding study in the rat, a decrease in the cellular components of the bone marrow, thymus and lymphoid follicles were observed at 50,000 ppm (3900 /4500 mg/kg/day (M/F) ), the highest dose tested. The NOAEL was 460/489 mg/kg/day (M/F). The potential for immunotoxicity is not of concern because the established chronic reference dose (cRfD) will be protective of these effects.

v. The dietary food exposure assessment utilizes proposed tolerance le vel or higher residues and 100% CT information for all commodities. By using these screening

- level assessments, chronic exposures/risks will not be underestimated.

VI I

. EXPOSURE

The request to approve of benzyl acetate is for post

- emergent use on gro wing crops. Human exposures to benzyl acetate applied as such are primarily expected to be those persons occupationally exposed by the inhalation and dermal routes of exposure by virtue of agriculturally related activities in volving pesticide application and related activities. Persons not occupationally exposed due to work activities may be exposed via the oral route of exposure to possible pesticide residues on treated crop commodities.

Dietary exposure is estimated using the Agency’s Dietary Exposur e Estimate Model (DEEM). Results are presented as an attachment and discussed separately. Results are discussed later

under

Occupational Exposure Since there is a large degree of uncertainty regarding the ultimate use(s) of benzyl acetate under

40 CFR 180.920, i.e., the submission is not specific as to target crops or methods of application

etc., the Agency conducted a screening level assessment of occupational exposures and risks. The Agency used assessment techniques developed by the Science Adviso ry Council on Exposure (ExpoSAC), Health Effects Division/OPP for the assessment of exposure to the amine oxides (EPA, 2009 ) . The algorithm used is:

Unit Exposure * Application Rate * Units Treated * Dermal Absorption ÷ Body Weight = Estimated Daily Dos e mg/lb inert lbs/unit acres treated ( 35 %) kg mg/kg/ day handled (acre) per day

For this assessment the Agency assumed dermal and inhala tion abs orption = 100%. Body weight is assumed to be 8 0 kg. Unit exposures are taken from the Occupational Pesticide Handler Unit Exposure Surrogate Reference Table (rev Mar. 2013)

. USEPA / Office ofPage 13 of 32

Pesticide Programs / Health Effects Division

. Ra tes of application and units treated are from ( Davis et al (2009) adjusted for 50 % inert in the final pesticide formulation

.

The Agency expresses Level of Concern (LOC) as Margin of Exposure (MOE) < 100. The MOE is the ratio of estimated daily dose to the toxicological endpoint used for risk assessment. Both are expressed as mg/kg/day and the result is unitless. Margins of exposure in the screening

level assessment are all > 100 except for mixer/loaders supporting high acreage aerial operations.

T able 2.0 ESTIMATED EXPOSURE/RISK APPLICATORS

Application

method Crop/Site Dermal

Unit Exposure Inhalation

Unit Exposure Rate of

Application Units

Treated Dermal

Dose Inhalation

Dose Dermal

MOE Inhalation

MOE

Aerial/liquid Corn 0.0021 0.0000049 5.2 1200 0. 06 0.00038 1833 289,000 Airblast/liquid Nut Trees 1.59 0.0047 0.8 40 0. 2 4 0.0019 458 57,894

Groundboom liquid Corn 0.0161 0.00034 5.2 200 0. 073 0.0044 1506 25,000

Convention to calculate exposure and risk. Unit Exposure (mg/lb inert handled) * Applic Rate (lb/A) * Units (A/day) * DA(%) ÷ body wt (80 kg) = Average Daily dermal or inhalation Dose (mg/kg/day) Margin of Exposure = NOAEL (mg/kg/day) ÷ Average Daily Dose (mg/kg/day). Rates of application taken from Davis et al 2009 (adjuste d for 50% per cent inert). Dermal Absorption = 35% Unit Exposures taken from Occupational Pesticide Handler Unit Exposure Surrogate Reference Table. U.S. Environmental Protection Agency, Office of Pesticide Programs. March 2013. Unpublished. U.E. for a pplicators wearing a single layer of work clothing (long pants, long

sleeved shirt, shoes plus socks) and protective gloves (pilots no gloves) and no respiratory protection.

Table 3.0 ESTIMATED EXPOSURE/RISK MIXER/LOADERS

Application

method Crop /Site Dermal

Unit Exposure Inhalation

Unit Exposure Rate of

Application Units

Treated Dermal

Dose Inhalation

Dose Dermal

MOE Inhalation

MOE

Aerial/liquid Corn 0.0376 0.000219 5.2 1200 1.03 0.0171 107 6,432 Airblast/liquid Nut Trees 0.8 40 0.0 0053 0.000 088 207547 1.25M

Groundboom liquid Corn 5.2 200 0. 17 0.0028 647 39,285

Residential Exposure As with the proposed agricultural uses, there is uncertainty regarding the possibility of “residential” use of this inert. The Agency conducted a “screening” level assessment of pesticide application activities known to result in high exposures, relatively speaking. The “home

- owner” pesticide handlers are assumed to wear short

- pants, short

- sleeved shirts and shoes plus sock s but no protective glove s. As with the agricultural assessment 100% absorption is assumed for the dermal and inhalation route of exposure. Thus the resulting estimates of risk are consideredPage 14 of 32

conservative, i.e. protective. The margins of exposure for residential handlers all exc eed 100. Residential exposures to benzyl acetate may also occur as the re sult of its use in cosmetics. Based on data considered in the evaluation of benzyl acetate as a fragrance in cosmetic products, ( MRID 494287 03 ), values for average daily exposure to benzyl acetate from cosmetic product use is included in the aggregate residential exposure assessment provided in section VIII below.

Table 4.0 Estimated Residential Pesticide Handler (Mixer/Loader/Applicator) Exposure and Risk

Mixer/loader/ applicato r Unit Exposure

mg/lb handled Application

Rate lb/ gal Units Treated

Acres/day Body weight

kg Average Daily

Dose mg/kg/day Margin of

Exposure

Hose end

sprayer Dermal 13.4

Inhal. 0.022 0. 0 105

lb/ gal 5 gal/day 80 kg Derm 0.0 0 308

Inhal 0.000 0 144 35

, 5 5 98

Pump

- up hand

- held

sprayer Dermal 63

Inhal 0.018 5 gal/day Derm 0. 0 144

Inhal 0.000 0 1 8 7 ,633

Backpack

sprayer Dermal 130

Inhal 0.14 5 gal/day Derm 0. 0 298

Inhal 0.000 0 92 3

, 6 7 9

Convention Used to Estimate Exposure = Unit Exposu re * Application rate * Units Treated ÷ Body weight

Factors used in residential exposure assessment:

Assumed 5

. 0% of formulation = inert. RD reviewed a number of residential product labels

– solvents typically comprised 1.0

- 5.0 % of the product formulat ion. Assumed product density 9.0 lb gal. Assumed typical rate of application 3 fl oz per formulation per gallon of finish spray Assumed 5 gallon spray applied per day to 1⁄2 A typical residence Body weight 80 kg from HED SOP

Dermal absorption = 35 %, inhalat ion absorption 100 % Unit Exposures from Standard Operating Procedures for Residential Pesticide Exposure Assessment Health Effects Division/ OPP January 2012. Toxicological Endpoint NOAEL 110 mg/kg/day Margin of Exposure = NOAEL mg/kg/day ÷ Average D aily Dose mg/kg/day . Since the same toxicological endpoint is used for dermal and inhalation and was identified from the same study, the dermal and inhalation exposures are summed prior to dividing into the NOAEL. HED SOP

9 lb/gallon formulation * 0. 0 5 (per cent inert) = 0.45 lb inert/gallon formulation

0.45 lb inert/gal formulation ÷ 128 fl oz /gal formulation = 0.0 0 35 lb inert/fl oz 3 fl oz per gallon spray * 0.035 lb inert/ fl oz = 0.105 lb inert per gallon For five gallons finish spray per day = 0. 0 525 lb inert per day

Margin of Exposure = NOAEL/Average Daily Exposure (dermal and inhalation exposures are summed prior to division into NOAEL).

Table 5.0 Estimated Residential Handler Indoor Exposures and Risks

Mixer/loader/ applicator Unit Expos ure

mg/lb handled Amount (lbs)

Handled/day Body weight

kg Average Daily

Dose mg/kg/day Margin of

ExposurePage 15 of 32

Mopping Dermal 71.6

Inhal. 2.38 0.0065 80 kg Derm 0.00 203

Inhal 0. 00019 54, 190

1.6M Wiping Dermal 2870

Inhal 67.3 0.00085 Derm 0.0 106

Inh al 0. 00072 10, 4 00

43,400 Trigger pump

spray Dermal 220

Inhal 2.4 0.05 Derm 0. 048

Inhal 0.0015 2 300

208,300

For the mopping scenario , it was assumed that 1 gallon of diluted solution is used. R D assumes that cleaning products are assumed to have the same density as water (8.34 lbs/gallon). While typical product concentrate directions indicate that 1 oz concentrated product per 1 gallon of water should be used, R D

followed the A ntimicrobials D ivision SOP and assumed a dilution factor of 2 oz concentrated product/128 oz (1 gallon) of water for the residential handler risk assessment to represent heavy

- duty cleaning formulations. Based on information from RD, the Agency assumed that the maximum percentage by weight of the inert ingredient in in door products would be 5.0 %.

1 gallon formulated pesticide solution * (8.34 lb/gal) * (5.0%) * (2 floz/128 floz dilution factor) =0.0065 lb inert in formu lated solution/ day

For wiping scenario

0.13 gallon formulated solution * (8.34 lb/gal) * (5.0%) * (2 floz/128 floz dilution factor) = 0.00085 lb inert in formulated solution/ day

For trigger pump spray

0.13 gallon formulated solution * (8.34 lb/gal) * (5.0%) = 0.00085 lb inert in formulated solution/ day

RESIDENTIAL and RECREATIONAL POST

- APPLICATION EXPOSURE

Table 6.0 Summary Recreational and Residential Post

- Application Exposures and Risks

Activity Exposure (Dose) mg inert./kg/day MOE

Toddler short

- term oral hand to mouth from contacting treated turf 0. 0036 30,555

Toddler oral object to mouth (turfgrass) 0.0 0023 478,000

Adult dermal post applic turf contact 0. 0172 6,395

Toddler dermal post applic turf

contact 0.033 3,333

Adult golfer post app turf contact 0. 0 00119 92,436

Child golfer post app turf contact 0. 00202 54,455

Toddler combined exposure 0. 037 = hand to mouth + object to 2,973Page 16 of 32

Table 6.0 Summary Recreational and Residential Post

- Application Exposures and Risks

mouth + turf post

- application

exposure

MOE = NOAEL  Dose. NOAEL = 110.0 mg inert/kg/day.

Adult and Adolescent Golfer Post

- Application Dermal Exposure may be estimated using the convention stated in Science Advisory Council for Exposure draft SOP regarding “Golfer Exposure Assessment For Adults and Children” (24 August 2000). The draft policy states that adult and adolescent golfer dermal post

- application exposure may be calcula ted as DE (t) (mg a.i./kg bw/day) =
(TTR (t) (μg/cm2)) * TC (cm2/hr) * hr/day  (CF (1000 μg/mg)) * BW (body weight (kg)) Where:

DE (t) = dermal exposure at time (t) attributable to golfing on previously treated turf (mg a.i./kg bw/day).

TTR (t) = turf transferable residue at time t (μg/cm2) TC = Transfer Coefficient (500 cm2/hr) hr = exposure period (4 hours) CF = conversion factor to change μg to mg. BW = body weight (kg) (80 kg for adult; adjusted (multiplied) by a factor of 1.7 for child golfers)

and where TTRt = AR * F * (1

- D) t * CF2 * CF3

AR = application rate (lb inert/acre); 0.021 lb ai/acre (taken from Table 4.0 above where

0. 0105 lb is applied to 1⁄2 acre) DA = Dermal absorption 35%

F = fract ion of inert available on turf/grass (unitless); 5% D = fraction of residue that dissipates daily (unitless); 10 t = postapplication day on which exposure is being assessed;

CF2 = weight unit conversion factor to convert the lbs ai in the application rate to use for the DFR or GR value (4.54E+8 μg/lb) CF3 = area unit conversion factor to convert the surface area units (acre) in the application rate to cm 2 for the TTR or GR value (2.47E

- 8 acre/cm 2 )

Therefore, 0.021 lb ai/A * 0.05 (%) * 4.54 8 μg/lb * 2.47

- 8 A/cm 2 = 0.136 μg/cm2

DE = 0.136 μg/cm2 * 500 cm2/hr * 4 hr/day * 0.001μg/mg * 0.35  80 kg bw = 0. 0 0119 mg inert/kg /day.

MOE for adult golfer is 110.0 mg a.i./kg bw/day  0. 0 0119 mg inert/kg bw/day = 92,436

.

The adult dose level is adjusted by a factor of 1.7 to estimate child golfer exposure therefore 0. 0 0119 mg inert./kg/day * 1.7 = 0. 00202 mg inert/kg/day.

MOE for child golfer is 110.0 mg inert/ kg/day  0. 00202 mg inert/day = 54,455

.Page 17 of 32

A MOE of 100 is adequate to protect golfers from dermal post

- application exposures to treated golf course turf. Inhalation exposure is believed to be negligible. The proposed use does not exceed the Agency’s lev el of concern.

Adult and toddler dermal post

- application exposure to treated residential turf is assessed using SOP (2.2) which indicates that

PDR = TTR * TC * ET * conversion factor μg to mg  kg bw where:

Potential Dose Rate (PDR)

Turf Transferabl e Residue (TTR) (0.588 μg/cm2) TC = Transfer Coefficient (TC)(14,500 cm2/hr for adult and 5,200 cm2/hr for child) ET = exposure time (hours/day) (2) CF = 0.001 mg/μg Dermal absorption = 35% Body weight = 80 kg for adult, 15 kg for toddler.

0.136 μg/cm 2 * 14,500 cm2/hr * 2 hr/day * 0.001 mg/μg * 0.35  80 kg bw = 0. 0172 mg inert/kg/day

for adult.

MOE = NOAEL  Dose and 110 mg a.i./kg bw/day  0. 0172 mg a.i./kg bw/day = 6,395

0.136 μg/cm2 * 5,200 cm2/hr * 2 hr/day * 0.001 mg/μg * 0.35  15 kg bw = 0.03 3 mg inert/kg/day for

toddler

MOE = NOAEL  Dose and 110 mg a.i./kg bw/day  0.033 = 3,333

.

Children’s short

- term oral hand

- to

- mouth exposure =

TTRt * SA * EX * FQ * ET * CF1 where:

PDR t = potential dose rate on day “t” (mg/kg bw/day)

TTRt = turf tra nsferable residue on day “t” (μg/cm2 turf); 0.00056 μg/cm2 SA = surface area of the hand (cm2/event; palmer surface area of 3 fingers = 20 cm2 EX = extraction factor from the hand by saliva = 50 % FQ = frequency of hand

- to

- mouth activity (events/h r); 20 events/hr ET = exposure time (hr/day); 2 hrs/day CF1 = weight unit conversion factor to convert μg in the TTR value to mg for the daily exposure (0.001mg/μg).

0.136 μg/cm2 * 20 cm2/event * 0.50 % extraction factor * 20 events/hr * 2 hr/day * 0.0 01 mg/μg  15 kg bw = 0. 0036 mg inert/kg/day.

MOE = NOAEL  Dose 110 mg inert/kg/day  0. 0036 mg inert/day = 30,555

.

Children’s object

- to

- mouth (turfgrass) = Grt * IgR * CF1Page 18 of 32

PDR t = potential dose rate on day “t” (mg/kg bw/day) GRt = grass (and plan t matter) residue on day “t”; 0.00056 μg/cm2 IgR = mouthing rate of grass (cm2/day); 25 cm2/day CF1 = weight unit conversion factor to convert μg units of residue on the grass to mg for the daily exposure (0.001mg/μg).

0.136 μg/cm2 * 25 cm2/day * 0.0 01mg/μg  15 kg bw = 0.0 0023 mg inert/kg/day

MOE = NOAEL  Dose and 110 mg inert/kg/day  0.0 0023 mg inert/kg/day = 478,000

.

VII I

. AGGREGATE EXPOSURE

In estimating total dietary (food and drinking water) exposures to benzyl acetate as an inert ingred ient in pesticide formulations applied to growing crops the Agency uses the Dietary Exposures Evaluation Model

– Food Commodity Intake Database ( DEEM

- FCID ) model for chronic dietary exposures which presented as Attachment One . The DEEM analysis was

conduc ted using the NOAEL noted earlier, 1 1 0 mg/kg/day , with the application of the standard uncertainty factors (100X)

.

1. In evaluating dietary exposure to the benzyl acetate , EPA considered exposure s under the petitioned for exemptions from the requirement of a tolerance. EPA assessed dietary exposures a s follows:

i. Acute exposure. No adverse effects attributable to a single exposure of benzyl acetate were seen in the toxicity databases. Therefore, an acute dietary risk assessment for benzyl acetate is not

necessary.

ii. Benzyl acetate was negative for carcinogenicity in mice and rats, therefore cancer risk

assessment was not conducted.

.

i ii. Chronic exposure. In conducting the chronic dietary exposure assessment, EPA used food consumption informatio n from the U.S. Department of Agriculture’s (USDA’s) 2003

- 2008 National Health and Nutrition Examination Survey, What We Eat in America (NHANES/WWEIA). As to residue levels in food, no residue data were submitted for acetic acid. In the absence of specif

ic residue data, EPA has developed an approach which uses surrogate information to derive upper bound exposure estimates for the subject inert ingredient. Upper bound exposure estimates are based on the highest tolerance for a given commodity from a list of high use insecticides, herbicides, and fungicides. A complete description of the general approach taken to assess inert ingredient risks in the absence of residue data is contained in the memorandum entitled ‘‘Alkyl Amines Polyalkoxylates (Cluster 4): Acute and Chronic Aggregate (Food and Drinking Water) Dietary Exposure and Risk Assessments for the Inerts,’’ (D361707, S. Piper, 2/25/09) and can be found at http://www.regulations.gov in docket ID number EPA

– HQ

– OPP

– 2008

– 0738.Page 19 of 32

In the dietary exposure assessment, the Agency assumed that the residue level of the inert ingredient would be no higher than the highest tolerance for a given commodity. Implicit in this assumption is that there would be similar rates of degradation (if any) between the active and inert ingredient and that the concentration of inert ingredient in the scenarios leading to these highest levels of tolerances would be no higher than the concentration of the active ingredient.

The Agency believes the assumptions used to estimate dietary exposures lead to an extremely conservative assessment of dietary risk due to a series of compounded conservatisms. First, assuming that the level of residue for an inert ingredient is equal to the level of residue for the active ingredient will overstate exposure. The concentrations of active ingredient in agricultural products are generally at least 50 percent of the product and often can be much higher. Further, pesticide products rarely have a single inert ingredient; rather there is general ly a combination of different inert ingredients used which additionally reduces the concentration of any single inert ingredient in the pesticide product in relation to that of the active ingredient. In the case of acetic

acid , EPA made a specific adjustm ent to the dietary exposure assessment to account for the use limitations of the amount of acetic acid that may be in pesticide formulations (limited to no more than 1200 ppm) present at the maximum limitation rather than at equal quantities with the activ e ingredient.

Second, the conservatism of this methodology is compounded by EPA’s decision to assume that, for each commodity, the active ingredient which will serve as a guide to the potential level of inert ingredient residues is the active ingredien t with the highest tolerance level. This assumption overstates residue values because it would be highly unlikely, given the high number of inert ingredients, that a single inert ingredient or class of ingredients would be present at the level of the

acti ve ingredient in the highest tolerance for every commodity. Finally, a third compounding conservatism is EPA’s assumption that all foods contain the inert ingredient at the highest tolerance level. In other words, EPA assumed 100 percent of all foods are treated with the inert ingredient at the rate and manner necessary to produce the highest residue legally possible for an active ingredient. In summary, EPA chose a very conservative method for estimating what level of inert residue could be on food, the n used this methodology to choose the highest possible

residue that could be found on food and assumed that all food contained this residue. No consideration was given to potential degradation between harvest and consumption even though monitoring data sh ows that tolerance level residues are typically one to two orders of magnitude higher than actual residues in food when distributed in commerce.

Accordingly, although sufficient information to quantify actual residue levels in food is not available, the compounding of these conservative assumptions will lead to a significant exaggeration of actual exposures. EPA does not believe that this approach underestimates exposure in the absence of residue data.

iii. Cancer. Benzyl acetate is not expected to b e carcinogenic since there was no evidence of carcinogenicity in rats and mice

, a cancer dietary exposure assessment was not performed.Page 20 of 32

2. Dietary exposure from drinking water. For the purpose of the screening level dietary risk assessment to support th is request for an exemption from the requirement of a tolerance for benzyl

acetate a conservative drinking water concentration value of 100 parts per billion (ppb) based on screening level modeling was used to assess the contribution to drinking water for chronic dietary

risk assessments for benzyl acetate . These values were directly entered into the dietary exposure

model. The DEEM model indicates the RfD (chronic population adjusted dose cPAD) is 1. 1 mg/kg/day

.

Aggregate Exposure for an Adult

0.189 m g/kg/day the estimated dietary exposure to the “Total U.S. Population” (adult) is

7.3 % of the cPAD 0.014 mg/kg/day exposure from non

- pesticidal use as a flavoring agent 0.12 mg/kg/day = exposure from use as a component of cosmetics

0. 0 298 mg/kg/day = hig hest residential handler exposure

0. 0172 mg/kg/day = post

- application dermal exposure to treated lawn/turf

0. 04 9 mg/kg/day = exposure from antimicrobial trigger pump spray

0. 0 0119 mg/kg/day = adult golfer post

- applications exposure.

0. 42 mg/kg/day thus 110 mg/kg/day ÷ 0. 42 mg/kg/day = a Margin of Exposure 260

Aggregate Exposure to Child age 1

- 2

0.691 mg/kg/day t he estimated dietary exposure to the most highly exposed subpopulation (toddlers 1

- 2 years) = 62.9 % of the cPAD

. 0. 014 mg/kg/day Toddlers are not expected to receive an equal exposure dose from use a flavoring agent. Lacking more refined estimates, the Agency uses the same amount as for adults.

0.033 mg/kg/day = post

- application dermal exposure to lawn/turf

0. 0036 mg/kg/day = hand to mouth exp osure 0.0 0023 mg/kg/day = object (turf grass) to mouth exposure

0.74 mg/kg/day thus 110 mg/kg/day ÷ 0.74 mg/kg/day = a Margin of Exposure 150

EPA determines whether acute and chronic pesticide exposures are safe by comparing aggregate exposure estimates to the aPAD and cPAD. The aPAD and cPAD represent the highest safe exposures, taking into account all appropriate safety factors (SFs). EPA calculates the aPAD and cPAD by dividing the POD by all applicable UFs. For linear cancer risks, EPA calculates the probability of additional cancer cases given the estimated aggregate exposure. Short

- , intermediate

- , and chronic

- term risks are evaluated by comparing the estimated aggregate food, water, and residential exposure to the POD to ensure that the MOE called for by the product of all applicable UFs is not exceeded. A n aggregate risk assessment was conducted including exposures from food, water, residential handler and residential post

- application.Page 21 of 32

1. Acute risk

. There was no hazard attributable to a single exposure seen in the toxicity

database for benzyl acetate . Therefore, benzyl acetate is not expected to pose an acute risk.

2. Chronic risk

. A chronic aggregate risk assessment takes into account exposure estimates from chronic dietary consumpt ion of food and drinking water using the exposure assumptions discussed above for chronic exposure. The chronic dietary exposure from food and water to benzyl acetate is 1 7.3 % of the cPAD for the U.S. population and 62.9 % of the cPAD for children

1

- 2 yr s old, the most highly exposed population subgroup.

3. Aggregate cancer risk for U.S. population . The EPA has not identified any concerns for carcinogenicity relating to benzyl acetate

.

4. Short

- term risk

. Short

- term aggregate exposure takes int o account short

- term residential exposure plus chronic exposure through use in pesticide formulations applied to growing crops. The estimated Margins of exposure for aggregate exposures are > 100 and do not exceed level of

concern.

I X . CUMULATIVE EXPOSU RE

Cumulative effects from substances with a common mechanism of toxicity . Section 408(b)(2)(D)(v) of FFDCA requires that, when considering whether to establish, modify, or revoke a tolerance, the Agency consider “available information” concerning the cu mulative effects of a particular pesticide's residues and “other substances that have a common mechanism of toxicity.”

EPA has not found benzyl acetate to share a common mechanism of toxicity with any other substances, and benzyl acetate does do not appe ar to produce a toxic metabolite produced by other substances. For the purposes of this tolerance action, therefore, EPA has assumed that benzyl

acetate do es not have a common mechanism of toxicity with other substances. For information regarding EPA's eff

orts to determine which chemicals have a common mechanism of toxicity and to evaluate the cumulative effects of such chemicals, see EPA's website at http://www.epa.gov/pesticides/cumulative

.

X. ENVI RONMENTAL FATE & EFFECTS

An acute toxicity study was conducted on the freshwater fish Oryzias latipes by Holcom b et al., (1995 ) . The study was conducted using continuous

- flow midi

- diluter exposure systems with Lake Superior water filtered and treated wit h ultraviolet light before it was heated to the testing temperature of 25° C. Water characteristics including dissolved oxygen, pH, hardness and alkalinity were measured during testing. The benzyl acetate duplicate agreement in test water was

97.4 %. Tw enty fish were exposed to concentrations of 0, 1.48, 2.45, 4.43, 8.81 or 19.8 mg/L benzyl acetate for 96 hours. The percentage survival at the two highest concentrations was zero and at the third highest (4.43 mg/L) survival was 80%. All lower concentrat ions had 100 %Page 22 of 32

survival through 96 hours. The LC 50 from this study was 4.00 mg/L (95 % C.I. 3.48

- 4.60). This

LC 50 for acute aquatic toxicity is generally categorized by the EPA/OPP as moderately toxic ( http://www.epa.gov/oppfed1ecorisk_ders/toers_analysis_eco.htm ).

The probability of rapid biodegradation of benzyl acetate was evaluated using EPI Suite TM 4.0 in the BIOWIN module. Benzyl acetate was estimated to undergo rapid b iodegradation and was categorized as readily biodegradable. The probability outputs from the Biowin 1 (linear model) and the Biowin 2 (non

- linear model) are used to determine the speed of biodegradation such that a biodegradation probability > 0.5 for bot h models indicates rapid biodegradation, while a biodegradation probability <0.5 for both models indicates “not rapid biodegradation” (EPA

- 2005). The benzyl acetate biodegradation probability in Biowin1 was 0.9783 and in Biowin2 0.9988, indicating rapid biodegradation. Similarly, probability outputs from the Biowin5 (MITI linea r model and Biowin6 (MITI non

- linear model are used to determine the ease of biodegradation such that a biodegradation probability >0.5 for both models indicates not readily degrada ble (EPA, 2005). Benzyl acetate biodegradation probability in Biowin5 was 0.5949 and in Biowin6 was 0.7562, indicating ready biodegradation. A biodegradation description of “weeks” or faster from Biowin3 in combination with biodegradation probability > 0 .5 from Biowin5 or Biowin6 provides a more reliable indicator of readily degradable for benzyl acetate.

X I

. ENVIRONMENTAL JUSTICE

Potential areas of environmental justice concerns, to the extent possible, were considered in this

human health risk assessm ent, in accordance with U.S. Executive Order 12898, "Federal Actions to Address Environmental Justice in Minority Populations and Low

- Income Populations," http://www.eh.doe.gov/oepa/gui dance/justice/eo12898.pdf ).

As a part of every pesticide risk assessment, OPP considers a large variety of consumer subgroups according to well

- established procedures. In line with OPP policy, HED estimates risks to population subgroups from pesticide e xposures that are based on patterns of that subgroup’s food and water consumption, and activities in and around the home that involve pesticide use in a residential setting. Extensive data on food consumption patterns are compiled by the USDA under

the Co ntinuing Survey of Food Intake by Individuals (CSFII) and are used in pesticide risk assessments for all registered food uses of a pesticide. These data are analyzed and categorized by subgroups based on age, season of the year, ethnic group, and region o f the country. Additionally, OPP is able to assess dietary exposure to smaller, specialized subgroups and exposure assessments are performed when conditions or circumstances warrant. Whenever appropriate, non

- dietary exposures based on home use of pestic ide products and associated risks for adult applicators and for toddlers, youths, and adults entering or playing on treated areas post

- application are evaluated. Further considerations are currently in development as OPP has committed resources and expert ise to the development of specialized software and models that consider exposure to bystanders and farm workers as well as lifestyle and traditional dietary patterns among specific subgroups.Page 23 of 32

XI I

. HUMAN STUDIES

This assessment relies in part on data from studies in which adult human subjects were intentionally exposed to a pesticide. These studies, listed below, have received the appropriate

ethical review for use in risk assessment.

The PHED Task Force, 1998. The Pesticide Handler Exposure Database (P HED), Version 1.1. Task Force members: Health Canada, U.S. Environmental Protection Agency, the California Department of Pesticide regulation, and the American Crop Protection Association; released August 1998.

XI I I

. RISK CHARACTERIZATION

Benzyl acet ate exhibits low levels of toxicity via the dermal route of exposure in rabbits and inhalation and oral routes of exposure in rats . It is mildly irritating to the skin and minimally irritating to the eyes of rabbits . It is not a skin sensitizer in guinea pigs

.

In a 13

- week feeding study in the rat, atrophic seminiferous tubules were observed in male rats at 12,500 ppm (900 mg/kg/day). The No Observed Adverse Effect Level (NOAEL) was identified as 6,250 ppm (460 mg/kg/day). In mice, following 13 weeks of exposure via the diet, decreased bodyweight and food consumption w ere observed at all doses. The LOAEL was 3,130 ppm (425 mg/kg/day). The NOAEL was not established.

In a developmental toxicity study, maternal and fetal toxicity were observed at 1000 mg/kg/day. Maternal toxicity was manifested as decreased bodyweight and fetal toxicity was manifested as reduced body weights, increased incidence of dilation of the renal pelvis and skeletal variations. Although qualitative fetal susceptibility is obse rved, fetal effects occur in the presence of maternal toxicity and a clear NOAEL of 500 mg/kg/day was established. Therefore, the established cRfD ( 1.10 mg/kg/day) will be protective of these effects.

The potential for benzyl acetate to be genotoxic w as evaluated in a battery of 10 in vivo mammalian genotoxicity studies. Two tests (a replicative DNA synthesis test at 1600 mg/kg/day and a Comet assay at 1200 and 1600 mg/kg) indicated positive potential to cause genotoxicity at the highest dose (1600 mg /kg). Eight studies were negative for the potential to be genotoxic. Based on the weight of the evidence, the Agency concludes the benzyl acetate is not genotoxic.

A series of twelve studies of the potential effects of benzyl acetate on several specie s of bacteria were reported. Two studies indicated positive effects and one study was reported as yielding equivocal results ( ( a ) Rec

- assay with B. subtilis at 20 μ L/disk and ( b ) a microscreen assay with E.

coli at 6.5 μ g/well). Ten studies were negative f or the effects of benzyl acetate to inducePage 24 of 32

mutagenic changes in bacteria. Based on the weight of the evidence the Agency concludes that benzyl acetate is negative for mutagenic effects.

Evidence of neurotoxicity and neuronal degeneration w ere identifie d in the 13 week NTP studies in rats and mice. Signs of neurotoxicity included tremors and ataxia that were associated with the degeneration of the glial cells in the cerebellum and hippocampus at the doses > 12,500 ppm (2,000/2,980 mg/kg/day). Since the se effects were induced at doses above the limit dose (1000 mg/kg/day) and the established c PAD was 1.10 mg/kg/day, these effect s will be protected. Therefore, the concern is low for these effects.

There is evidence that benzyl acetate suppresses immune function in mammalian systems in the rat

occurr ing only at a dose that was lethal and w e ll above the limit dose. In the 13 week feeding study in the rat, a decrease in the cellular components of the bone marrow, thymus and lymphoid

follicles were observed at 50,000 ppm (3900 mg/kg/day for males and 4500 mg/kg/day for females), the highest dose tested. The NOAEL for this study was 12,500 ppm (900 mg/kg/day). The potential for immunotoxicity is not of concern because the established chronic population adju sted dose (c PAD ) will be protective of these effects.

Carcinogenicity studies were conducted by NTP in rats and mice. Benzyl acetate was administered via the diet to rats and mice at doses up to 12,000 ppm (510/575 mg/kg/day, male/female). Toxicity wa s not observed in rats at any dose . In mice, males and females exhibited reduced body weight throughout the entire study.

The No Observed Adverse Effect Level (NOAEL) selected for use in risk assessment is taken from

the two

- year feeding study conducted by the NTP (1993 ) using B6C3F 1 mice. The NOAEL of 110 mg/kg/day is based on decreased body weight. Throughout the 2

- year study, the mean body weights were 9 % to 14 % lower than those of the control animals.

The request to establish a tolerance exemption for residues of benzyl acetate on growing crop primarily expected for use in liquid insecticides however some herbicides may be applied to growing crops (harvest aides etc). Human exposures to benzyl acetate applied as such are expected for those persons occupationally involved in commercial agricultural activities. Exposure is expected by the inhalation and dermal routes. Persons not occupationally exposed due to work activities may be exposed via the oral route of exposure to possible pesticide residue s on treated crop commodities.

Dietary exposure is estimated using the Agency’s Dietary Exposure Estimate Model (DEEM). Results are presented as an attachment and discussed separately. Estimated exposures do not exceed the Agency’s level of concern. Th e estimated dietary exposure to the “Total U.S. Population” (adult) is 0.189 mg/kg/day which equates to 17.3 % of the RfD. The estimated dietary exposure to the most highly exposed subpopulation (toddlers 1

- 2 years) is

0. 692 mg/kg/day which equates to 62. 9 % of the RfD. Dietary Margins of Exposure are:Page 25 of 32

541 for adults (U.S. Population) and 156 for toddlers 1

- 2 years (most highly exposed subpopulation).

Screening level non

- occupational (i.e., residential) exposure assessments result in Margins of Expo s ur e greater than 100. A screening level occupational exposure assessment indicates all estimated Margins of Exposure are > 100 except for mixer/loaders supporting high acreage aerial applications.

Estimates of aggregate exposure are Margins of Exposure 26 0 for adults and 150 for children 1

- 2 years. These Margins of Exposure do not exceed the Agency’s level of concern . Further, the estimates of exposure and risk are based on very conservative assumptions

. Moreover, the Agency considers the possibility hig hly remote for simultaneous exposures to all the application and post

application scenarios described. Further, the highly conservative dietary estimate of exposure indicates the dietary exposure to children 1

- 2 years (the most highly exposed subpopulatio n) equal s 62.9% of the cPAD is being protective and therefore not exceeding the Agency’s level of

concern

.

Benzyl acetate is described as moderately toxic to some freshwater organisms. Extensive modeling using the EPISuite TM environmental model system indicates benzyl acetate is readily biodegradable and not likely to bioaccumulate.

Based on the weight of the evidence

, the toxicological database for benzyl acetate generally

indicat es low levels of toxicity, (including mutagenicity and carcinogenicity ) , dietary exposures

not exceeding levels of concern, low occupational and residential exposures and risk and the indication of minimal adverse environmental effects, the Agency approves of the requested exemption from the requirement of a tolerance for be nzyl acetate as an inert ingredient (solvent) in pesticide formulations applied to growing crops under 40 CFR 180.920

.Page 26 of 32

REFERENCES

21 CFR 172.515. Food Additives Permitted for Direct Addition to Food for Hu m an Consumption, Subpa rt F. Flavoring Agents and Related Substances. Synthetic Flavoring Substances and Adjuvants.

Adams, T., S. Cohen, J. Doull, V. Feron, J. Goodman, L Marnett, I. Munro, P. Portoghese, R. Smith, W. Waddell and B. Wagner. 2005. The FEMA GRAS assessment of benzyl derivatives used as flavor ingredients. Food an d Chemical Toxicology. 43: 1207

- 1240 cited in MRID 49428701). MRID 49428703.

Belsito, D., D. Bickers, M. Bruze, P. Calow, M. Dagli, A. Fryer, H. Greim, Y. Miyachi, J. Saurat and I. Sipes. (2012). A Toxicological and Dermatological Assessment of Aryl Alkyl Alcohol Simple Acid Ester Derivatives when Used As Fragrance Ingredients. Food and Chemical Toxicol.

50: S269

- S313.

Bronaugh, R.L. and H.I. Maibach. 1985. Percutanious absorption of nitroa romatic compounds in

vivo and in vitro studies in the human and monkey. J. Invest. Derm. 84. 180. MRID 49428708.

Bronaugh, R.L., R.C. Wester, J. Bucks, H.I. Maibach and R. Sarason. 1990. In vivo percutaneous absorption of fragrance ingredients in rhes us monkeys and humans. Food and Chemical Toxicology. 28 (5) 369

- 373. MRID 49428703.

Davis, D., B

- H. Chin, A. LaMay, M. Lloyd, C. Olinger, S. Piper. 2009. Alkyl Alcohol Alkoxylate Phosphate and Sulfate Derivatives (AAAPDs and AAASDs

– JITF CST 2 Inert Ingredients). Human Health Assessment to Spport Proposed Exemption from the Requirement of a Tolerance When Used as Inert Ingredients in Pesticide Formulations. CASRN 9046

- 01

- 9. PC Code 911339. Unpublished Memorandum 8 June 2009 to K. Leifer U.S. EPA.

Environmental Protection Agency (EPA). 2009. “ 900439 , (C10

- 16); C10

- C18

- Alkyldimethylamine oxides; Human Health Risk Assessment and Ecological Effects Assessment to Support Proposed Exemption from the Requirement of a Tolerance When Used as Inert Ingre dients in Pesticide Formulations. ” Memorandum L. Austin to P. Shah.

International Agency for Research on Cancer (IARC) Monograph, Benzyl Acetate Volume 71 p

1255. http://monogra phs.iarc.fr/ENG/Monographs/vol71/mono71

- 65.pdf

.

Holcombe, G., D. Benoit, D. Hammermeister, E. Leonard and R. Johnson. 1995. Acute and Lon

Term Effects of Nine Chemicals on the Japanese Medaka ( Orysias latipes ). Archives

Environmental Contamin. & Tox icol. 28: 287

- 297.Page 27 of 32

McGinty, D., D. Vitale, C. Letizia, A. Api. 2012. Fragrance material review on benzyl acetate. Food and Chemical Toxicology. 50: 5363

- 5384. MRID 49428711.

National Library of Medicine (NLM), H o useh o ld products Database, Benzyl A cetate entry
assessed 4/14. http://householdproducts.nlm.nih.gov/cgi

bin/household/brands?tbl+chem&id=307 7&query=benzyl+acetate&searchas=TblChemicals1&pro

dcat=all

.

National Toxicology Program (NTP). 1993. Toxicology and Carcinogenesis studies of Benzyl Acetate (CAS No. 140

- 11

- 4) in F344/N Rats and B6c3f Mice (Feed Studies). National Toxicology Program Te chnical Report Series No. 431. http://ntp.niehs.nih.gov/ntp/htdocs/lt_rpts/tr431.pdf

. MRID 49428712.

World Health Organization (WHO), 1996. Food Additive Series 37 Benzyl Acetate, Ben zyl Alcohol, Benzaldehyde and Benzoic Acid and its Salts. World Health Organization, Geneva 1996. http://www.inchem.org/documents/jecfa/jecmono/v37je01.htm

.

MRID 49428701. 2014 . Petition Proposing An Exemption From the Requirement of a Tolerance for Residues of: Benzyl Acetate (CAS No. 140

- 11

- 4) under 40 CFR § 180.920. Author B. E. Mileson, Huntsman Corporation.

MRID 49428702. 2005. The FEMA GRAS Assessment of Benzyl Deriv atives Used as Flavor Ingredients. Author T.B. Adams et al., Huntsman Corporation.

MRID 49428704. 1999. Benzylacetate Monograph. International Agency for Research on Cancer World Health Organization. Submitted for Huntsman Corporation.

MRID 49428705 / 1996/ Benzyl Acetate, Benzyl Alcohol, Benzaldehyde and Benzoic Acid and Its Salts. Author E. Vavasour. Chemical Health Hazard Assessment Division, Food Directorate Health Protection Branch, Health Canada.

MRID 49428706. 1986. Studies on Benzyl A cetate. I. Effect of dose Size and vehicle on the Plasma Pharmacokinetics and Metabolism of [Methylene

- 14C] Benzyl Acetate in the Rat. Authors M.A.J. Chidgey and J. Caldwell. Department of Pharmacology, St. Mary’s Hospital Medical School, London, Engla nd. Submitted for Huntsman Corporation.

MRID 49428707. 1986. Studies on Benzyl Acetate. II. Use of Specific Metabolic Inhibitors to Define the Pathway Leading to the formation of Benzylmercapturic Acid in the Rat. Authors M.A.J. Chidgey, J.F. Kennedy and J. Caldwell. Department of Pharmacology, St. Mary’s Hospital Medical School, London, England. Submitted for Huntsman Corporation.Page 28 of 32

MRID 49428708. 1987. Studies on Benzyl Acetate. III. The Percutaneous Absorption and Disposition of [Methylene

14 C] Benzyl Acetate in the Rat. Authors M.A.J. Chidgey, J.F. Kennedy and J. Caldwell. Department of Pharmacology, St. Mary’s Hospital Medical School, London, England. Submitted for Huntsman Corporation.

MRID 49428709. 1995. Acute and Long

- Term Effects of Nine Chemicals on the Japanese Medaka ( Oryzias latipes ). Authors, G.W. Holcombe, D.A., Benoit, D.E. Hammermeister, E.N.

Leonard and R.D. Johnson. U.S> Environmental Protection Agency/Environmental Research Laboratory

- Duluth Minnesota. Submitted for Hunt sman Corporation.

MRID 49428710. 2013. Household Products Database: Benzyl acetate. U.S. Department of Health and Human Services. Submitted for Huntsman Corporation.Page 29 of 32

ATTACHMENT ONE

Evaluation Copy Ver. 4.02, 05

- 10

- c DEEM

- FCID Chronic ana lysis for INERTS

- BENZYLACETATE NHANES 2005

- 2010

2

- day

Residue file name:

K: \ BenzylAcetate \ BenzylAcetate INERTS_57ACTIVE_100PPBH2OREV.R1 Adjustment factor #2 NOT u sed. Analysis Date 10

- 20

- 2015/11:23:59 Residue file dated: 10

- 20

- 2015/11:22:43 Reference dose (RfD, Chronic) = 1.1 mg/kg bw/day COMMENT 1: Inert 57 active ingredients + drinking water (100ppb)

=========================================================== ==========

==========

Total exposure by population subgroup

-------------------------------------------------------------------------------

Total Exposure

----------------------------------- Population mg/kg Percent of Subgroup body wt/day Rfd

-------------------------------------- ------------- ---------------- Total US Po pulation 0.189889 17.3% Hispanic 0.202223 18.4%

Non

- Hisp

- Whi te 0.187701 17.1%

Non

- Hisp

- Bl ack 0.173910 15.8%

Non

- Hisp

- Oth er 0.218231 19.8% Nursing Inf

ants 0.216739 19.7%

Non

- Nursing Infants 0.439736 40.0%

Female 13+ P REG 0.169286 15.4%

Children 1

- 6 0.538692 49.0%

Children 7

- 12 0.226058 20.6%

Male 13

- 19 0.13760 0 12.5%

Female 13

- 1 9/NP 0.134955 12.3%

Male 20+ 0.142454 13.0%

Female 20+/ NP 0.156073 14.2%

Seniors 55+ 0.154296 14.0%

All Infants 0.369591 33.6%

Female 13

- 50 0.147618 13.4%

Children 1

- 2 0.691658 62.9%

Children 3

- 5 0.481575 43.8%

Children 6

- 12 0.248815 22.6%Page 30 of 32

Youth 13

- 19 0.136273 12.4%

Adults 20

- 49 0.146019 13.3%

Adults 50

- 99 0.155151 14.1%

Female 13

- 49 0.147643 13.4%Page 31 of 32

ATTACHMENT TWO

SUMMARY OF in vivo Mammalian Genotoxicity Studies ( from McGinty et al 2012; MRID 49428701)

Method Target Dosage Results

Unscheduled DNA F344 Rat Hepatocytes 50, 200 or 1000 mg/kg Negative Synthesis

Alkaline elution F344 Rat Pancreatic cells 150, 500 or 1500 mg/kg Negative Analysis

Comet assay Wistar Rats 1200 or 1600 mg/kg Positive

Unscheduled DNA Fischer 344 Rats 1000 mg/kg Negative Synthesis

Sister Chromatid BC6F31 mouse bone marrow IP (5/dose) 325 to 1700 mg/kg Negative Exchange

Chromosomal aberrations BC6F31 mouse bone marrow IP (5/dose) 325 to 1700 mg/kg Negative

Micronucleus assay Male BC63F1 mice 312.5, 625 or 1250 mg/kg x 3 Negative

Micronucleus assay Male and female B6C3F1 mice 470, 938, 1875, 3750 or 7500 mg/k g Negative

In diet for 13 weeks

Replicative DNA B6C3F1 mice hepatocytes 800 mg/kg Negative Synthesis test 1600 mg/kg Positive

Comet assay ddy mice 1200 and 1600 mg/kg PositivePage 32 of 32

Summary of Bacterial Studies with Benzyl Acetate ( from McGinty et al 2012; MRID 49428701)

Test System Species Concentration Results

Ames test S. typhimurium TA 100 50,000 μ g/plate Negative

Modified Ames S. typhimurium 33, 100, 333, 1,000, Negative

or 10,000 μ g/plate

Ames test S. typhimurium 0.045 μ g/plate Negative

Ames test S. typhimurium Up to 4.5 μ g/plate Negative

Modified Ames test and plate S. typhimurium 5, 50 or 500 μ g/plate Negative incorporation assay with UV

radiation

umu test S. typhimurium up to 5000 μ g/plate Negative

Rec

- assay B. subtilis 20 μ L/disk Positive

E. coli 250

- 2000 μ /plate Negative

2000

- 8000 μ g/mL Negative

Rec

- assay B. subtilis 21 μ L/disk Negative

SOS chromotest E. coli 0.2

- 50 μ g/disk Negative

Microscreen assay E. coli 6.5 μ g/well Positive

Mutatox bioluminescence assay P. phosphoreum N/A equivocal

Mitotic chromosome loss Yeast strain D61.M 520

- 1817 μ g.mL Negative

255

- 917 μ g.mL in Negative proprionitrile