i

EPA REGISTRATION DIVISION COMPANY NOTICE OF FILING FOR PESTICIDE PETITIONS 

PUBLISHED IN THE FEDERAL REGISTER 

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

Jeneil Biosurfactant Company

[IN-10846]

EPA has received a pesticide petition (IN-10846) from Technology Sciences Group Inc. 

(1150 18th Street, Suite 1000 Washington, DC 20036) on behalf of Jeneil Biosurfactant 

Company, 400 N. Dekora Woods Blvd. Saukville, WI 53080 proposing, pursuant to section 

408(d) of the Federal Food, Drug, and Cosmetic Act (FFDCA), 21 U.S.C. 346a(d), to amend 40 CFR 

part 180.

(Options (pick one)

2. to establish an exemption from the requirement of a tolerance for

Isobutyl acetate, CAS #110-19-0, when used as an inert solvent applied to growing 

crops or harvested crops under 40 CFR 180.910 in pesticide formulations. EPA has determined 

that the petition contains data or information regarding the elements set forth in section 408 

(d)(2) of FDDCA; however, EPA has not fully evaluated the sufficiency of the submitted data at 

this time or whether the data supports granting of the petition. Additional data may be needed 

before EPA rules on the petition.

A. Residue Chemistry NA remove

1. Plant metabolism.

2. Analytical method. NA remove

3. Magnitude of residues. NA remove
2

B. Toxicological Profile Isobutyl acetate, which has been identified as one of the primary 

fragrance compounds in bananas and is also a product of yeast fermentation processes, 

metabolizes readily to innocuous, endogenous products. Therefore, oral toxicity resulting from 

the use of isobutyl acetate as an inert ingredient in pesticides is not expected to be significant. 

To further evaluate the toxicity of isobutyl acetate the European Chemicals Bureau (ECB) IUCLID 

dataset on isobutyl acetate acid was consulted. In addition, data on isobutyl alcohol from the US 

EPA’s IRIS database and the European Chemical Agency’s dossier are referenced. Isobutyl 

alcohol is one of the primary metabolic products of isobutyl acetate, and therefore these data 

can be bridged to assess the toxicity of the acetate form.

1. Acute toxicity. Acute toxicity tests categorize isobutyl acetate into acute toxicity 

category IV for oral, inhalation, and dermal exposure. Oral LD50 values range from 4763 mg/kg 

bw in rabbits to 13,400 mg/kg bw in rats. Acute inhalation toxicity is likewise low, with a 6-hour 

LC50 of >13.24 mg/L calculated for rats. The dermal LD50 in rabbits was determined to be >17,400 

mg/kg bw in the 24-hour Draize cuff method. Another acute dermal study derived an LD50 of 

>5000 mg/kg bw in rabbits.

2. Genotoxicty. Isobutyl acetate did not induce reverse mutations in Salmonella 

typhimurium strains TA98, TA100, TA1535, TA1537, and TA1538 at concentrations as high as 

5000 μg/mL in the absence or presence of metabolic activation.

3. Reproductive and developmental toxicity. In two prenatal developmental toxicity 

inhalation studies, mated female Wistar rats and Himalayan rabbits were exposed to isobutanol 

(99.8%) at vapor concentrations of 0, 151, 758 or 3030 ppm (0, 500, 2,500 or 10,000 mg/m3) for 

six hours/day. The rats were dosed on post-coitum days 6-15. No mortality or significant 

differences in clinical signs, body weight development, or gross pathology between controls and 

treated groups were observed. No effects on development were noted. The maternal and 

developmental rat NOAELs were 3030 ppm. Rabbits (15/dose) were dosed on postinsemination days 7-19. High-dose rabbit dams had a slightly lower body weight gain, 

particularly during the first phase of exposure. The maternal NOAEL for rabbits was 758 ppm. 

Fetuses exhibited no signs of developmental changes in response to isobutanol. Therefore, the 

rabbit developmental NOAEL was 3030 ppm, the highest dose.

In a 2-generation reproductive inhalation study isobutanol (99.9%) was administered to 30 

Crl:CD(SD)IGS BR rats/sex/dose by whole body exposure at 0, 500, 1000 and 2500 ppm (0, 1476, 

2952, and 7380 mg/m3) for 6 hours per day 7 days per week. Exposure began at the 

commencement of the study for the F0 generation, and on postnatal day 28 for the F1 

generation. It ended after weaning of the pups (F0 generation postnatal day 28, F1 generation 

postnatal day 21). For dams, exposure was discontinued after day 20 of gestation through 

lactation day 5. No exposure-related effects were observed on F0 and F1 parental survival or on 

F0 and F1 reproductive performance, body weights, food consumption and food efficiency in 
3

males or females. There were no significant spermatogenic or macroscopic findings, changes in 

primordial follicle or corpora lutea counts, changes in mean organ weights or histopathologic 

lesions in the F0 or F1 rats at any dose. Pup endpoints were not significant for either F1 or F2 

generations, including survival, general physical condition, and mean body weights. There were 

no macroscopic findings, changes in mean organ weights, or maturation of F1 pups. The NOAEL 

of isobutanol for parental systemic, reproductive and neonatal toxicity is 2500 ppm (7380 

mg/m3, the maximum applied dose) in males and females of the F0, F1 and F2 

generations (ECHA 2014b). The study was rated “reliable without restriction” by the European 

Chemicals Agency (ECHA).

4. Subchronic toxicity. No repeat-dose toxicity studies were identified for isobutyl 

acetate. However, based on the toxicity profile of isobutyl alcohol (isobutanol), which is one of 

the primary metabolic products of isobutyl acetate, and based on the widespread occurrence of 

isobutyl acetate in food, both as a synthetic food additive and natural component of many plant 

species and food-related fermentation processes, the long-term toxicity of isobutyl acetate is 

not expected to be significant. A 90-day oral rodent toxicity study on isobutyl alcohol served as 

the scientific basis for US EPA’s oral Reference Dose (RfD) for that compound. Isobutyl alcohol 

was administered to 30 Sprague-Dawley (Crj: CD(SD)) rats/sex/dose by gavage for 90 days at 

dose levels of 0, 100, 316, or 1000 mg/kg bw/day. Treatment related effects were seen only at 

1000 mg/kg bw/day, and included hypoactivity, which was significant during week one and 

decreased markedly after week 4, and lower body weight gain (18% below that of control rats) 

in males during week one. Food consumption averages were below those of the controls for 

both males and females during weeks 1 and 2. Clinical signs and effects on body weight and 

food consumption were all transient and were considered as adaptive and therefore as not 

adverse. Serum potassium levels were 11-15% lower in both sexes compared to controls on day 

29 , but group mean serum potassium concentrations were similar for treated and control 

groups at the final clinical pathologic evaluation. No other significant differences between dosed 

groups and controls were found for ophthalmoscopic examination, hematology, urinalysis, 

organ weights, gross pathology and histopathology. The NOEL was determined to be 316 mg/kg 

bw/day and the NOAEL was 1000 mg/kg/day. U.S. EPA derived an oral RfD of 0.3 mg/kg/day for 

isobutyl alcohol by applying uncertainty factors totaling 1000 to this NOEL.

In another study, Wistar rats (10/sex/group) were exposed to isobutanol in drinking water at 

concentrations of 0, 1000, 4000, or 16,000 ppm, equivalent to approximately 0, 80, 340, or 1450 

mg/kg bw/day isobutanol actually ingested. The 90-day study was carried out under OECD Test 

Guideline 408 and included full hematology, urinalysis, and ophthalmological endpoints as well 

as post-sacrifice histological exam. Body weight, food and water consumption, and clinical signs 

of toxicity were monitored throughout the study. There were no significant dose-dependent 

signs of toxicity in any of these parameters. Study authors concluded that the NOAEL for 

isobutanol was the highest dose, 1450 mg/kg/day.

In a 90-day guideline study (Guideline: 82-7, Subdivision F (Neurotoxicity Screening Battery)), 

99% pure isobutanol was administered by dynamic whole-body exposure to 10-20 Sprague-
4

Dawley rats/sex/dose at 0, 250, 1000, or 2500 ppm for 6 hours per day, 5 days/week (~ 0.75, 

3.0, 7.5 mg/L). In high dose females, total erythrocyte count, hemoglobin, and hematocrit were 

slightly elevated compared to controls (p ≤ 0.05). The European Chemicals Agency (ECHA), which 

reviewed the study, considered this to be not biologically significant. No differences were found 

in body weight, food consumption, ophthalmoscopic examination, clinical observation, clinical 

chemistry, neurobehavioral observations, organ weights, gross pathology, and histopathology. 

The NOAEL for repeat-dose effects was the highest dose, 2500 ppm.

5. Chronic toxicity. No chronic studies were identified for isobutyl acetate or its 

metabolites.

6. Animal metabolism. Metabolism of aliphatic esters such as isobutyl acetate proceeds 

rapidly through hydrolysis to form an alcohol and carboxylic acid. These are reactions of the 

carboxylesterases or esterases, which predominate in hepatocytes but are present in most 

tissues throughout the body, including small intestine, colon, kidney, trachea and lungs. The 

initial metabolites of isobutyl acetate are isobutyl alcohol and acetic acid. Isobutyl alcohol is 

oxidized via alcohol dehydrogenase and aldehyde dehydrogenase to isobutyric acid. The acids 

will be conjugated and excreted or will undergo β-oxidation in the fatty acid metabolic pathway. 

The metabolic products are endogenous in mammals and innocuous at levels produced from the 

typical diet.

7. Metabolite toxicology. NA Remove

8. Endocrine disruption. Toxicity data related to endocrine disruption were not identified 

in the isobutyl acetate database. As the scientific knowledge develops, screening of additional 

compounds may be added to the Endocrine Disruptor Screening Program (EDSP). When 

additional screening and/or testing is conducted, isobutyl acetate may be the focus of screening 

and/or testing to better characterize effects related to endocrine disruption.

C. Aggregate Exposure

1. Dietary exposure. The estimated dietary exposure to isobutyl acetate was determined 

using methods to estimate chronic dietary exposure for a generic inert ingredient. This 

assessment considers drinking water and crop-specific residues from pre-harvest applications of 

agricultural insecticides, herbicides and fungicides, assuming the highest established tolerance 

level residue for each commodity. The assessment assumes that the inert ingredient is used on 

all crops. The inert ingredient is assumed to be present in all commodities treated with 57 of 

the most significant active ingredients at the maximum tolerance level as identified by the U.S. 

EPA for the default assessment. Chronic dietary exposure estimates were derived for the 

general US population and sub-groups of the population using the Dietary Exposure Evaluation 

Model, DEEMTM. Assuming all crops are treated with formulated products containing 10% 

isobutyl acetate by weight, the estimated chronic exposure for the total US population is 0.038

mg/kg/day, 7.7% of the chronic Population Adjusted Dose (cPAD). Children age 1 to 2 years old 

have the highest estimated exposure at 0.141 mg/kg/day, or 28.5% of the cPAD.
5

i. Food. Dietary exposures of concern are not anticipated for isobutyl acetate due to its

lack of persistence in the environment and low general toxicity. A significant source of dietary 

isobutyl acetate comes from natural food sources and its use as a food additive. JECFA (1999) 

estimated the current level of intake of isobutyl acetate as a food-flavoring agent to be 1300 

μg/day in the U.S., equivalent to approximately 0.0186 mg/kg bw/day and reported that there 

was no safety concern at the current level of intake.

ii. Drinking water. Isobutyl acetate exposure via drinking water from use as an inert 

ingredient is not expected to be significant. Isobutyl acetate is inherently biodegradable and will 

volatilize relatively quickly from water. Exposure via the drinking water was estimated in the 

DEEM dietary exposure assessment assuming a concentration of 100 ppb.

2. Non-dietary exposure. Residential exposures of concern are not anticipated for 

isobutyl acetate due to its inherent ability to biodegrade in the environment, low projected uselevel in pesticides, and low general toxicity.

D. Cumulative Effects

Section 408(b)(2)(D) (9v) of the FFDCA requires that, when considering whether to 

establish, modify, or revoke a tolerance, the Agency consider “available information” concerning 

the cumulative effects of a particular pesticide’s residues and “other substances that have a 

common mechanism of toxicity.” 

To our knowledge there are no available data or other reliable information that suggests any

toxic effects produced by isobutyl acetate would be cumulative with those of any other chemical 

compounds. EPA has not made a common mechanism of toxicity finding as to isobutyl acetate 

and other compounds. Isobutyl acetate does not produce toxic metabolites in common with 

other substances of potential concern. For the purpose of the tolerance exemption proposed, it 

is assumed that isobutyl acetate does not share a common mechanism of toxicity with other 

substances

E. Safety Determination

1. U.S. population. Taking into consideration all available information on isobutyl 

acetate, there is a reasonable certainty that no harm to any population subgroup will result 

from dietary and other nonoccupational exposures to isobutyl acetate when used as an inert 

ingredient in an inert mixture intended for use in pesticide formulations. Adjusting for relative 

molecular weight, EPA’s oral RfD of 0.3 mg/kg/day for isobutanol was used to derive an RfD for 

isobutyl acetate of 0.495 mg/kg/day, which added uncertainty factors totaling 1000 for 

interspecies extrapolation, intraspecies variability, and for extrapolating subchronic exposure to 

chronic exposure. The chronic Population Adjusted Dose (cPAD) was determined to be the same 

as the RfD because no effects of isobutanol on development or reproduction were observed, 

suggesting that isobutyl acetate too would not increase susceptibility of sensitive populations. 

Therefore, the oral RfD and cPAD are both 0.495 mg/kg/day. For the total US population, the 
6

estimated chronic dietary exposure from food and drinking water for isobutyl acetate, 

calculated as 10% of all agricultural formulations, is 7.7% of the cPAD, well below any level of 

potential concern.

2. Infants and children. An evaluation of susceptibility and uncertainty issues associated 

with isobutyl acetate has been performed and there is a reasonable certainty that no harm to 

infants and children will result from the use of isobutyl acetate as an inert ingredient in pesticide 

products. The structurally similar and metabolically related compound isobutyl alcohol was not a 

developmental toxin to rats or rabbits following inhalation. For this reason, a safety factor 

analysis has not been implemented to assess risk and the additional tenfold safety factor for the 

protection of infants and children is also unnecessary. The estimated chronic dietary exposure 

from food and drinking water was highest (28.5% of cPAD) for children aged 1 to 2 years old. 

This estimated exposure to isobutyl acetate is well below any level of potential concern. 

F. International Tolerances

NA Remove