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 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 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 (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 were 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 the 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 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. 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 studies indicated positive effects and one study was reported as yielding 

equivocal results ((a) Rec-assay with B. subtilis at 20μL/disk and (b) a microscreen assay with E. 

coli at 6.5 μg/well). 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.

Evidence of neurotoxicity and neuronal degeneration 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 

(2,000/2,980 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 effects 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 occurring 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 lymphoid 

follicles were observed at 50,000 ppm (3900 mg/kg/day for males and 4500 mg/kg/day for 
Page 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 the 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 studies 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 concluded 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 female mice.

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, MRID 49428712) using B6C3F1

mice. The NOAEL of 110 mg/kg/day is based on decreased body weight 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 Maibach, 1985, MRID 49428708; Bronaugh et al, 1990, MRID 

49470803). 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 (applicator 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 Model (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 cRfD. The 

estimated dietary exposure to the most highly exposed subpopulation (children 1-2 years old) is

0.691 mg/kg/day which equates to 62.9% of the cRfD. 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 residential 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 indicates 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 EPISuiteTM 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 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 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 18th Street, NW, Suite 1000, Washington, D.C. 20036) 

on behalf of the Huntsman 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-harvest, 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 humans 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 food 

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 reconfirmed the group 

as “GRAS”. 

Approximately one-third of the current dietary exposure to benzyl acetate occurs from naturallyoccurring benzyl acetate in a variety of fruit, vegetables and other foods and the other two-thirds 
Page 5 of 32

is from use as a flavoring ingredient (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 

μg/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

LogKOW -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 1,605a

3,100b

Log KOC 2.059c

1.944d

Henry’s Law Constant atm-m3/mole @ 25°C( 2.302 x 10 -005

Probability of Rapid Biodegradation (BIOWIN) 0.9783e

0.9988f

3.0295 days-weeksg

0.5949h

0.7562i

Ready Biodegradability Yes

BCF (L/kg wet-wt) 9.124

Volatilization from Water Half-Life (hr) 64.71j

809.1k

Fugacity Model Compartment/Mass Amount % Air: 2.86

Water: 26.9

Soil: 70

Sediment: 0.173

Fugacity Model Compartment/Half-Life (hr) Air: 40.2

Water: 360

Soil: 720

Sediment: 3.24 x 103

aWSKOW modeled result gBiowin3 (Ultimate survey model)

bExperimental database match identified in the model hBiowin5 (MITI linear model)

cMCI method result iBiowin6 (MITI non-linear model)

dKOW method result jModel River

eBiowin 1 (linear model) kModel Lake
Page 6 of 32

fBiowin 2 (non-linear model)

Physical/Chemical properties taken from MRID 49428701.

IV. TOXICOLOGICAL SUMMARY

Acute Oral Toxicity – Benzyl acetate exhibits low acute oral toxicity to rats. The oral LD50 for 

the rat is 2,490 to 3,690 mg/kg (WHO, 1996. MRID 49428701). 

Acute Dermal Toxicity – The dermal LD50 in rabbits is > 5,000 mg/kg (McGinty et al., 2012. 

MRID 49428701).

Acute Inhalation Toxicity – Rats exposed (whole body) to 766 μg/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 was at the maximum 

attainable vapor concentration. The LC50 was > 766 μg/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 dose 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 dermal 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 mildly 

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 once 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 National 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, 3,130, 

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 group. Feed consumption by exposed rats, 

except the 25,000 and 50,000 ppm males 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 may have 

been due to toxicity or decreased palatability. The reduction in body weight coincided with the 

decrease in feed consumption. Tremors and ataxia occurred only in the 50,000 ppm rats. These 

findings were first observed on day 15 in nine males and six females and continued until the end 

of the study. Cholesterol levels at 12,500 ppm and 25,000 ppm and triglyceride levels in 25,000

ppm in females were lower (14-20%) than those in the controls. However these changes are not 

toxicologically significant. Mild to moderate aspermatogenesis was observed in two 50,000 ppm 

males and atrophy of seminiferous tubules were observed 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 renal tubule epithelium and degeneration and sarcolemma 

nuclear hyperplasia of the tongue and skeletal muscles occurred in most male and female 50,000 

ppm rats. The 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 

B6C3F1 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 mg/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 those of the controls and the mean body 

weight gains decreased with increased exposure level. Feed consumption by 3,130 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 occurred 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. The 
Page 8 of 32

LOAEL was 3,130 ppm (425 mg/kg/day) based on decreased body weights and feed 

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 not 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 group 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 ribs, 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 were 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, 3900/4500 mg/kg/day for males and females, respectively) in

the 13 week NTP study. Signs of neurotoxicity included tremors and ataxia that were associated 

with the degeneration of the glial cells in the cerebellum and hippocampus. However, these 

effects were 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 observed 
Page 9 of 32

in females at > 12,500 ppm (2,000/2,980 mg/kg/day). Since the effects were induced 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 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 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 reference dose (cRfD) will be protective of these effects.

Carcinogenicity

Rat

The National Toxicology Program (NTP) conducted a 2-year feeding study of benzyl acetate on 

rats (NTP, 1993; MRID 49428712). Groups of 60 male and female F344/N rats received benzyl 

acetate in the diet at concentrations 0, 3,000, 6,000, or 12,000 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. 

Dietary 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 or 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 no 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 feeding study was conducted by the NTP (1993; MRID 49428712) using B6C3F1

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 

females. 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 and 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 severities 
Page 10 of 32

of non-neoplastic nasal lesions occurred in the most posterior portions of the nasal cavity in all 

exposed groups. The lesions occurred in the majority of the exposed mice and consisted of 

atrophy and degeneration primarily of the olfactory epithelium, cystic hyperplasia of the nasal 

submucosal glands, pigmentation of the mucosal epithelium and exudate accumulation. There is 

no evidence of carcinogenic activity of benzyl acetate in male or female B6C3F1 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 B6C3F1 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 fore-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 acetate 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 neoplasms 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 B6C3F1 mice were given 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 

metabolite 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 frequency of dosing had any effect on the absorption, metabolism or excretion of 

benzyl acetate. 

The percutaneous absorption and disposition of [methylene14C] benzyl acetate was studied in 

F344 rats by applying benzyl 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 Maibach, 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 for benzyl acetate is 110 mg/kg/day from the NTP 2-year carcinogenicity

study in mice (MRID 49428701) where 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 inhalation 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 effects 

on body weight were 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 infants 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. Qualitative fetal susceptibility was observed in the developmental study in rats. Maternal 

(decrease in bodyweight) and fetal (reduced bodyweights, increased incidence of dilation of the 

renal pelvis and skeletal variations) toxicity were 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 be protective of these 

effects. The potential for reproduction toxicity was observed in the 13 weeks dietary study in 

rats. Atrophy of seminiferous tubules was observed in males at 12,500 ppm (900 mg/kg/day). 

However, the concern for reproduction toxicity 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 developmental toxicity study in rats.

iii. Neurotoxicity studies were not available for review. Evidence of neurotoxicity and 

neuronal degeneration was identified in the 13 week NTP study. Signs of neurotoxicity included 

tremors and ataxia that were associated 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 the 
Page 12 of 32

limit 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 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 (3900/4500 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 level or higher residues 

and 100% CT information for all commodities. By using these screening-level assessments, 

chronic exposures/risks will not be underestimated.

VII. EXPOSURE 

The request to approve of benzyl acetate is for post-emergent use on growing 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 involving 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 Exposure 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 Advisory 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 Dose

mg/lb inert lbs/unit acres treated (35 %) kg mg/kg/day

handled (acre) per day

For this assessment the Agency assumed dermal and inhalation absorption = 100%. Body 

weight is assumed to be 80 kg. Unit exposures are taken from the Occupational Pesticide 

Handler Unit Exposure Surrogate Reference Table (rev Mar. 2013). USEPA / Office of 
Page 13 of 32

Pesticide Programs / Health Effects Division. Rates 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. 

Table 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 0.24 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 (adjusted 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 applicators wearing a single layer of work clothing (long pants, longsleeved 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 0.00053 0.000088 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 socks but 

no protective gloves. As with the agricultural assessment 100% absorption is assumed for the 

dermal and inhalation route of exposure. Thus the resulting estimates of risk are considered 
Page 14 of 32

conservative, i.e. protective. The margins of exposure for residential handlers all exceed 100. 

Residential exposures to benzyl acetate may also occur as the result of its use in cosmetics. 

Based on data considered in the evaluation of benzyl acetate as a fragrance in cosmetic products, 

(MRID 49428703), 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/

applicator

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

0.0105

lb/gal

5 gal/day 80 kg Derm 0.00308

Inhal 0.0000144

35,5598

Pump-up 

hand-held 

sprayer

Dermal 63

Inhal 0.018

5 gal/day Derm 0.0144

Inhal 0.000018

7,633

Backpack

sprayer

Dermal 130

Inhal 0.14

5 gal/day Derm 0.0298

Inhal 0.000092

3,679

Convention Used to Estimate Exposure = 

Unit Exposure * 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 formulation.

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 %, inhalation 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 Daily 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 * 0.05 (per cent inert) = 0.45 lb inert/gallon formulation

0.45 lb inert/gal formulation ÷ 128 fl oz /gal formulation = 0.0035 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 = 0.0525 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 Exposure

mg/lb handled

Amount (lbs)

Handled/day

Body weight

kg

Average Daily

Dose mg/kg/day

Margin of 

Exposure
Page 15 of 32

Mopping Dermal 71.6

Inhal. 2.38

0.0065 80 kg Derm 0.00203

Inhal 0.00019

54,190

1.6M

Wiping Dermal 2870

Inhal 67.3

0.00085 Derm 0.0106

Inhal 0.00072

10,400

43,400

Trigger pump 

spray

Dermal 220

Inhal 2.4

0.05 Derm 0.048

Inhal 0.0015

2300

208,300

For the mopping scenario, it was assumed that 1 gallon of diluted solution is used. RD 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, RD 

followed the Antimicrobials Division 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 indoor 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 formulated 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)

0.00023 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 0.000119 92,436

Child golfer post app turf contact 0.00202 54,455

Toddler combined exposure 0.037 = hand to mouth + object to 2,973
Page 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 calculated 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 = fraction 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 cm2for the TTR or GR value (2.47E-8 acre/cm2) 

Therefore, 0.021 lb ai/A * 0.05 (%) * 4.548 μg/lb * 2.47-8 A/cm2 = 0.136 μg/cm2

DE = 0.136 μg/cm2 * 500 cm2/hr * 4 hr/day * 0.001μg/mg * 0.35  80 kg bw = 0.00119 mg inert/kg 

/day. 

MOE for adult golfer is 110.0 mg a.i./kg bw/day  0.00119 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 0.00119

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 level 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 Transferable 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/cm2 * 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 = 0.033 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:

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

TTRt = turf transferable 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/hr); 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 * 0.001 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 * CF1
Page 18 of 32

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

GRt = grass (and plant 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 * 0.001mg/μg  15 kg bw = 0.00023 mg inert/kg/day

MOE = NOAEL  Dose and 110 mg inert/kg/day  0.00023 mg inert/kg/day = 478,000.

VIII. AGGREGATE EXPOSURE 

In estimating total dietary (food and drinking water) exposures to benzyl acetate as an inert 

ingredient 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 

conducted using the NOAEL noted earlier, 110 mg/kg/day, with the application of the standard 

uncertainty factors (100X). 

1. In evaluating dietary exposure to the benzyl acetate, EPA considered exposures under the 

petitioned for exemptions from the requirement of a tolerance. EPA assessed dietary exposures as 

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..

iii. Chronic exposure. In conducting the chronic dietary exposure assessment, EPA used food 

consumption information 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 specific 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 generally 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 adjustment 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 active 

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 ingredient 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 

active 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, then 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 shows 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 be 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 this 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 mg/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

0.0298 mg/kg/day = highest residential handler exposure

0.0172 mg/kg/day = post-application dermal exposure to treated lawn/turf

0.049 mg/kg/day = exposure from antimicrobial trigger pump spray

0.00119mg/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 the 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 exposure

0.00023mg/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. An 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 consumption 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 17.3 % of the cPAD for the U.S. population and 62.9 % of the cPAD for children 

1-2 yrs 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 into 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.

IX. CUMULATIVE EXPOSURE

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 cumulative 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 appear to produce a toxic metabolite produced by other 

substances. For the purposes of this tolerance action, therefore, EPA has assumed that benzyl 

acetate does not have a common mechanism of toxicity with other substances. For information 

regarding EPA's efforts 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. ENVIRONMENTAL FATE & EFFECTS

An acute toxicity study was conducted on the freshwater fish Oryzias latipes by Holcomb et al., 

(1995). The study was conducted using continuous-flow midi-diluter exposure systems with Lake 

Superior water filtered and treated with 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 %. Twenty 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 concentrations had 100 % 
Page 22 of 32

survival through 96 hours. The LC50 from this study was 4.00 mg/L (95 % C.I. 3.48-4.60). This 

LC50 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 SuiteTM 4.0 in 

the BIOWIN module. Benzyl acetate was estimated to undergo rapid biodegradation 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 both 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 linear

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 degradable (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.

XI. ENVIRONMENTAL JUSTICE

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

human health risk assessment, 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/guidance/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 exposures 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 Continuing 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 of 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 pesticide 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 expertise 

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

XII. 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 (PHED), 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.

XIII. RISK CHARACTERIZATION 

Benzyl acetate 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 were 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 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 was 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 species 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 20μL/disk and (b) a microscreen assay with E. 

coli at 6.5 μg/well). Ten studies were negative for the effects of benzyl acetate to induce 
Page 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 were 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 

(2,000/2,980 mg/kg/day). Since these effects were induced at doses above the limit dose (1000 

mg/kg/day) and the established cPAD was 1.10 mg/kg/day, these effects 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 

occurring 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 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 

adjusted dose (cPAD) 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 was 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 B6C3F1 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 residues 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. The 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 

Exposure 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 260 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 highly remote for simultaneous exposures to all the application and postapplication scenarios described. Further, the highly conservative dietary estimate of exposure 

indicates the dietary exposure to children 1-2 years (the most highly exposed subpopulation) 

equals 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 EPISuiteTM 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 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 benzyl 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 Human Consumption, 

Subpart 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 and 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 nitroaromatic 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 rhesus 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 

Ingredients in Pesticide Formulations.” Memorandum L. Austin to P. Shah.

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

1255. http://monographs.iarc.fr/ENG/Monographs/vol71/mono71-65.pdf. 

Holcombe, G., D. Benoit, D. Hammermeister, E. Leonard and R. Johnson. 1995. Acute and LonTerm Effects of Nine Chemicals on the Japanese Medaka (Orysias latipes). Archives 

Environmental Contamin. & Toxicol. 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), Household products Database, Benzyl Acetate entry 

assessed 4/14. http://householdproducts.nlm.nih.gov/cgibin/household/brands?tbl+chem&id=3077&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 Technical 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, Benzyl 

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 Derivatives 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 Acetate. 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, England. 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 [Methylene14C]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 Huntsman 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 analysis for INERTS- BENZYLACETATE NHANES 2005-2010 

2-day

Residue file name: 

K:\BenzylAcetate\BenzylAcetateINERTS_57ACTIVE_100PPBH2OREV.R1 

Adjustment factor #2 NOT used.

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 Population 0.189889 17.3%

Hispanic 0.202223 18.4%

Non-Hisp-White 0.187701 17.1%

Non-Hisp-Black 0.173910 15.8%

Non-Hisp-Other 0.218231 19.8%

Nursing Infants 0.216739 19.7%

Non-Nursing Infants 0.439736 40.0%

Female 13+ PREG 0.169286 15.4%

Children 1-6 0.538692 49.0%

Children 7-12 0.226058 20.6%

Male 13-19 0.137600 12.5%

Female 13-19/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/kg 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 Positive
Page 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 μg/plate Negative

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

or 10,000 μg/plate

Ames test S. typhimurium 0.045 μg/plate Negative

Ames test S. typhimurium Up to 4.5 μg/plate Negative

Modified Ames test and plate S. typhimurium 5, 50 or 500 μg/plate Negative

 incorporation assay with UV

 radiation

umu test S. typhimurium up to 5000 μg/plate Negative

Rec-assay B. subtilis 20 μL/disk Positive

E. coli 250-2000 μ/plate Negative

 2000-8000 μg/mL Negative

Rec-assay B. subtilis 21 μL/disk Negative

SOS chromotest E. coli 0.2-50 μg/disk Negative

Microscreen assay E. coli 6.5 μg/well Positive

Mutatox bioluminescence assay P. phosphoreum N/A equivocal

Mitotic chromosome loss Yeast strain D61.M 520-1817 μg.mL Negative

255-917 μg.mL in Negative

proprionitrile