This report contains the collective views of an international group of experts and does not necessarily represent the decisions or the stated policy of the United Nations Environment Programme, the International Labour Organization, or the World Health Organization.

Concise International Chemical Assessment Document 64

First draft prepared by Philip Copestake, Toxicology Advice & Consulting Ltd, Surrey, United Kingdom; and Mr Heath Malcolm, Centre for Ecology & Hydrology, Monks Wood, United Kingdom

Published under the joint sponsorship of the United Nations Environment Programme, the International Labour Organization, and the World Health Organization, and produced within the framework of the Inter-Organization Programme for the Sound Management of Chemicals.

World Health Organization

Geneva, 2005

The International Programme on Chemical Safety (IPCS), established in 1980, is a joint venture of the United Nations Environment Programme (UNEP), the International Labour Organization (ILO), and the World Health Organization (WHO). The overall objectives of the IPCS are to establish the scientific basis for assessment of the risk to human health and the environment from exposure to chemicals, through international peer review processes, as a prerequisite for the promotion of chemical safety, and to provide technical assistance in strengthening national capacities for the sound management of chemicals.

The Inter-Organization Programme for the Sound Management of Chemicals (IOMC) was established in 1995 by UNEP, ILO, the Food and Agriculture Organization of the United Nations, WHO, the United Nations Industrial Development Organization, the United Nations Institute for Training and Research, and the Organisation for Economic Co-operation and Development (Participating Organizations), following recommendations made by the 1992 UN Conference on Environment and Development to strengthen cooperation and increase coordination in the field of chemical safety. The purpose of the IOMC is to promote coordination of the policies and activities pursued by the Participating Organizations, jointly or separately, to achieve the sound management of chemicals in relation to human health and the environment.

WHO Library Cataloguing-in-Publication Data

Butyl acetates.

(Concise international chemical assessment document ; 64)

1.Acetates - toxicity 2.Acetates - adverse effects 3.Risk assessment 4.Environmental exposure I.International Programme on Chemical Safety II.Series.

ISBN 92 4 153064 2           (LC/NLM Classification: QD 305.A2)

ISSN 1020-6167

©World Health Organization 2005

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Technically and linguistically edited by Marla Sheffer, Ottawa, Canada, and printed by Wissenchaftliche Verlagsgesellschaft mbH, Stuttgart, Germany

FOREWORD

Concise International Chemical Assessment Documents (CICADs) are the latest in a family of publications from the International Programme on Chemical Safety (IPCS) — a cooperative programme of the World Health Organization (WHO), the International Labour Organization (ILO), and the United Nations Environment Programme (UNEP). CICADs join the Environmental Health Criteria documents (EHCs) as authoritative documents on the risk assessment of chemicals.

International Chemical Safety Cards on the relevant chemical(s) are attached at the end of the CICAD, to provide the reader with concise information on the protection of human health and on emergency action. They are produced in a separate peer-reviewed procedure at IPCS. They may be complemented by information from IPCS Poison Information Monographs (PIM), similarly produced separately from the CICAD process.

CICADs are concise documents that provide summaries of the relevant scientific information concerning the potential effects of chemicals upon human health and/or the environment. They are usually based on selected national or regional evaluation documents or on existing EHCs. Before acceptance for publication as CICADs by IPCS, these documents undergo extensive peer review by internationally selected experts to ensure their completeness, accuracy in the way in which the original data are represented, and the validity of the conclusions drawn.

The primary objective of CICADs is characterization of hazard and dose–response from exposure to a chemical. CICADs are not a summary of all available data on a particular chemical; rather, they include only that information considered critical for characterization of the risk posed by the chemical. The critical studies are, however, presented in sufficient detail to support the conclusions drawn. For additional information, the reader should consult the identified source documents upon which the CICAD has been based.

Risks to human health and the environment will vary considerably depending upon the type and extent of exposure. Responsible authorities are strongly encouraged to characterize risk on the basis of locally measured or predicted exposure scenarios. To assist the reader, examples of exposure estimation and risk characterization are provided in CICADs, whenever possible. These examples cannot be considered as representing all possible exposure situations, but are provided as guidance only. The reader is referred to EHC 170.¹

While every effort is made to ensure that CICADs represent the current status of knowledge, new information is being developed constantly. Unless otherwise stated, CICADs are based on a search of the scientific literature to the date shown in the executive summary. In the event that a reader becomes aware of new information that would change the conclusions drawn in a CICAD, the reader is requested to contact IPCS to inform it of the new information.

Procedures

The flow chart shows the procedures followed to produce a CICAD. These procedures are designed to take advantage of the expertise that exists around the world — expertise that is required to produce the high-quality evaluations of toxicological, exposure, and other data that are necessary for assessing risks to human health and/or the environment. The IPCS Risk Assessment Steering Group advises the Coordinator, IPCS, on the selection of chemicals for an IPCS risk assessment based on the following criteria:

• there is the probability of exposure; and/or
• there is significant toxicity/ecotoxicity.

Thus, it is typical of a priority chemical that

• it is of transboundary concern;
• it is of concern to a range of countries (developed, developing, and those with economies in transition) for possible risk management;
• there is significant international trade;
• it has high production volume;
• it has dispersive use.

The Steering Group will also advise IPCS on the appropriate form of the document (i.e., a standard CICAD or a de novo CICAD) and which institution bears the responsibility of the document production, as well as on the type and extent of the international peer review.

The first draft is usually based on an existing national, regional, or international review. When no appropriate source document is available, a CICAD may be produced de novo. Authors of the first draft are usually, but not necessarily, from the institution that developed the original review. A standard outline has been developed to encourage consistency in form. The first draft undergoes primary review by IPCS to ensure that it meets the specified criteria for CICADs.

The second stage involves international peer review by scientists known for their particular expertise and by scientists selected from an international roster compiled by IPCS through recommendations from IPCS national Contact Points and from IPCS Participating Institutions. Adequate time is allowed for the selected experts to undertake a thorough review. Authors are required to take reviewers’ comments into account and revise their draft, if necessary. The resulting second draft is submitted to a Final Review Board together with the reviewers’ comments. At any stage in the international review process, a consultative group may be necessary to address specific areas of the science. When a CICAD is prepared de novo, a consultative group is normally convened.

The CICAD Final Review Board has several important functions:

• to ensure that each CICAD has been subjected to an appropriate and thorough peer review;
• to verify that the peer reviewers’ comments have been addressed appropriately;
• to provide guidance to those responsible for the preparation of CICADs on how to resolve any remaining issues if, in the opinion of the Board, the author has not adequately addressed all comments of the reviewers; and
• to approve CICADs as international assessments.

Board members serve in their personal capacity, not as representatives of any organization, government, or industry. They are selected because of their expertise in human and environmental toxicology or because of their experience in the regulation of chemicals. Boards are chosen according to the range of expertise required for a meeting and the need for balanced geographic representation.

Board members, authors, reviewers, consultants, and advisers who participate in the preparation of a CICAD are required to declare any real or potential conflict of interest in relation to the subjects under discussion at any stage of the process. Representatives of nongovernmental organizations may be invited to observe the proceedings of the Final Review Board. Observers may participate in Board discussions only at the invitation of the Chairperson, and they may not participate in the final decision-making process.

This CICAD² on n-butyl, isobutyl, sec-butyl, and tert-butyl acetates was prepared by Toxicology Advice & Consulting Ltd and the Centre for Ecology & Hydrology. The health effects sections were based on the Dutch Expert Committee on Occupational Standards and Swedish Criteria Group for Occupational Standards Basis for an Occupational Standard (Stouten & Bogaerts, 2002). Data identified as of September 2000 were considered in this source document. A comprehensive literature search of several online databases was conducted by Toxicology Advice & Consulting Ltd in January 2004 to identify any references published subsequent to those incorporated in the source document. The environmental and ecotoxicological sections were prepared by the Centre for Ecology & Hydrology from a review of the literature. Information on the nature of the peer review and the availability of the source document is presented in Appendix 2. Information on the peer review of this CICAD is presented in Appendix 3. This CICAD was considered and approved as an international assessment at a meeting of the Final Review Board, held in Hanoi, Viet Nam, on 28 September – 1 October 2004. Participants at the Final Review Board meeting are presented in Appendix 4. The International Chemical Safety Cards for n-butyl acetate (IPCS, 2003a), isobutyl acetate (IPCS, 2003b), sec-butyl acetate (IPCS, 2003c),and tert-butyl acetate (IPCS, 2002), produced by the International Programme on Chemical Safety in a separate, peer-reviewed process, have also been reproduced in this document.

The butyl acetate isomers n-butyl acetate (CAS No. 123-86-4), isobutyl acetate (CAS No. 110-19-0), sec-butyl acetate (CAS No. 105-46-4), and tert-butyl acetate (CAS No. 540-88-5) are colourless, flammable liquids with fruity odours.

Butyl acetates can occur naturally, and they are present in various plant tissues. They may be released to the environment from industrial plants during their manufacture and use, as well as following their use as solvents in products such as lacquers, inks, coatings, and adhesives. n-Butyl acetate is used as a food flavorant and in materials for food contact use. Butyl acetates may also be formed in the atmosphere as a product of the photochemical oxidation of other chemicals.

Butyl acetates released to the environment are likely to volatilize to the atmosphere, where they will undergo photochemical oxidation reactions with hydroxyl radicals and chlorine atoms. Butyl acetates in solution will undergo hydrolysis reactions, at a rate determined by the pH of the solution. Butyl acetates are readily biodegradable. Their physicochemical properties suggest that butyl acetates will not bind to soil or be bioaccumulated.

Butyl acetates have been detected in river water, but the concentrations were not quantified. They have also been detected in air samples from industrial and chemical waste sites at concentrations up to 4.8 µg/m³. Exposure of the general population may occur from domestic sources, with n-butyl acetate concentrations up to 23 µg/m³ reported in household air. Occupational exposure to butyl acetate particles and vapour may occur in workplaces involving painting, printing, lacquering, or glueing. Mean occupational air concentrations as measured by personal air sampling ranged up to 413 mg/m³.

It is expected that the butyl acetates are readily absorbed by the respiratory tract, the skin, and the gastrointestinal tract, although no published quantitative data were identified. n-Butyl acetate, isobutyl acetate, and sec-butyl acetate may be readily hydrolysed to acetic acid and their respective alcohols (n-butanol, isobutanol, and sec-butanol) in the blood, liver, small intestine, and respiratory tract. tert-Butyl acetate is less readily hydrolysed, with about 20% of the inhaled isomer being metabolized by a different pathway involving hydroxylation to produce 2-hydroxyisopropyl acetate. Where appropriate, data on the alcohols relevant to an assessment of toxic hazard and risk of butyl acetates have been included in this CICAD. n-Butyl acetate is probably excreted via exhaled air and urine both as the unchanged compound and as metabolites after transformation in the body. Humans exposed to atmospheres containing n-butyl acetate at a concentration of 200 mg/m³ were reported to excrete 50% of the inhaled compound in the exhaled air.

Data on the acute inhalation toxicity of n-butyl acetate are highly inconsistent, with LC₅₀ values ranging from 740 to above 45 000 mg/m³. The explanation for the inconsistent results is not known. However, the results of a recent well designed and performed experiment indicate that the toxicity of n-butyl acetate following a single 4-h inhalation is low, with no deaths occurring at exposures up to approximately 45 000 mg/m³. Additionally, n-butyl acetate has low acute toxicity by the oral and dermal routes. Oral LD₅₀ values in male and female rats were 13.1 and 11.0 g/kg body weight, respectively, whereas no deaths occurred in rabbits exposed by the dermal route to 14.4 g/kg body weight. Data (where available) on the other isomers indicate low toxicity by the inhalation, oral, and dermal routes.

Most results indicate that n-butyl, isobutyl, and tert-butyl acetates are, at most, only slightly irritating to the skin and eyes, although there is some indication of more severe irritation with certain exposure conditions. No data on irritation were identified for sec-butyl acetate. n-Butyl acetate and isobutyl acetate have been tested for skin sensitization potential, with negative results.

Published data on systemic toxicity following repeated exposure are limited to n-butyl acetate. The principal effect observed following inhalation exposure was a reduction in activity levels at 7200 mg/m³ and above, with a NOAEC of 2400 mg/m³. However, a 13-week neurotoxicity study in which rats were exposed by inhalation to atmospheres containing up to 14 000 mg/m³ found no evidence of neurotoxicity in functional observational battery, motor activity, and scheduled-controlled operant behaviour tests or on microscopic examination of nervous system tissues.

Only limited studies (with only one tested concentration) are available on the reproductive and developmental toxicity of n-butyl acetate. Although there were signs of developmental toxicity reported, maternal toxicity was also present. Data from a developmental toxicity study with the major metabolite, n-butanol, suggest that it is not a developmental toxin. No data were identified on the other butyl acetate isomers. Studies with the key metabolites isobutanol and sec-butanol indicate lack of specific reproductive or developmental toxicity.

None of the butyl acetate isomers has been tested in long-term carcinogenicity studies. Results (where available) from genotoxicity studies, however, indicate a lack of activity. Although the metabolite tert-butanol has given some evidence of carcinogenicity in rats and mice, genotoxicity assays with this compound again failed to show any activity.

Human studies indicate that n-butyl acetate exposure via inhalation may be slightly irritating to the eyes, nose, and throat. Sensitivity to odour occurs at concentrations several orders of magnitude lower than levels at which nose and throat irritation are reported. Isobutyl acetate (2% in petrolatum) was not irritating when applied as a 48-h covered patch. No, or only very limited, data concerning effects on humans were available on the other isomers.

Based on the limited data set for n-butyl acetate, a tolerable concentration of 0.4 mg/m³ has been derived. This is based on results from the 13-week inhalation study in rats providing the lowest NOAEC. An uncertainty factor of 1000 is used, allowing for interspecies extrapolation, intraspecies variability, and extrapolation from medium-term to long-term exposure. The only available study in which representative levels of n-butyl acetate in households were identified reported values up to 0.02 mg/m³, which is 20 times less than the tolerable concentration. Occupational exposure levels, however, may exceed this tolerable concentration.

Acute toxicity data suggest that butyl acetate has moderate to low toxicity to aquatic organisms. An EC₅₀ value of 675 mg/litre was reported for growth of green algae exposed to n-butyl acetate for 72 h. Twenty-four-hour LC₅₀/EC₅₀ values for aquatic invertebrates exposed to n-butyl acetate and isobutyl acetate were 72.8–205 mg/litre and 250–1200 mg/litre, respectively. Ninety-six-hour LC₅₀ values for fish ranged from 18 to 185 mg/litre for n-butyl acetate. Forty-eight-hour LC₅₀ values for fish exposed to isobutyl acetate ranged from 71 to 141 mg/litre. NOEC values for growth of lettuce exposed to tert-butyl acetate were 100 mg/litre (14-day NOEC in soil) and 32 mg/litre (16-day NOEC in hydroponic solution).

The butyl acetate isomers n-butyl acetate, isobutyl acetate, sec-butyl acetate, and tert-butyl acetate are colourless, flammable liquids with fruity odours. The relative molecular mass of each isomer is 116.2. Some physicochemical properties for the butyl acetate isomers are outlined in Table 1. An odour threshold value for nbutyl acetate is reported as 1.9 mg/m³ (Amoore & Hautala, 1983). Other odour threshold values reported for butyl acetates are 0.031 mg/m³ (Kruize, 1988) and 0.92 mg/m³ (Devos et al., 1990), although it is not clear to which isomers these refer.

Table 1: Physical and chemical properties.

^a Exists in D- and L-isomeric forms.

The structural formulae of the four butyl acetate isomers are given below:

n-butyl acetate

isobutyl acetate

sec-butyl acetate

tert-butyl acetate

The conversion factors³ for the butyl acetate isomers in air (at 20 °C, 101.3 kPa) are:

1 ppm = 4.83 mg/m³

1 mg/m³ = 0.207 ppm

In this CICAD, data on butyl alcohols (relative molecular mass 74.12) that are relevant to an assessment of the toxic hazard and risk of butyl acetates have been included, where appropriate. The conversion factors for the butyl alcohols in air (at 20 °C and 101.3 kPa) are as follows:

1 ppm = 3.08 mg/m³

1 mg/m³ = 0.325 ppm

Technical grades of butyl acetates contain butyl alcohol as an impurity, and small amounts of water may also be present (Syracuse Research Corp., 1979). Commercial grades that are in current use are better defined and of higher purity than those used in the early 1930s, when studies on the toxicity of these esters began (Zaleski, 1992).

In cosmetic grades of n-butyl acetate, lesser amounts of n-butyl alcohol and isobutyl alcohol and traces of n-propyl acetate and isobutyl acetate are present, with a maximum of 10% for the sum of all possible impurities (Toy, 1989).

Methods are available for measuring butyl acetates in environmental samples (NIOSH, 1994). Ten litres of air are sampled on a solid sorbent tube (coconut shell charcoal) and desorbed with carbon disulfide. Aliquots are analysed by gas chromatography equipped with a flame ionization detector. For 10-litre air samples, the method is applicable for n-butyl acetate, isobutyl acetate, sec-butyl acetate, and tert-butyl acetate at concentration ranges of 352–1475 mg/m³, 306–1280 mg/m³, 478–2005 mg/m³, and 424–1780 mg/m³, respectively.

The use of diffusive samples in monitoring butyl acetate vapours in indoor/workplace air has been reported (De Bortoli et al., 1987; Gentry & Walsh, 1987; Kristensson & Beving, 1987; Sala, 1987).

Butyl acetates can be determined by infrared and ultraviolet spectroscopy, gas chromatography, gas chromatography/mass spectrometry, and headspace gas chromatography (Toy, 1989; Weller & Wolf, 1989).

Butyl acetate recovery rates were measured in two types of canister used to sample atmospheric air for analysis of volatile organic compounds, including butyl acetate, at different relative humidities. Recoveries after 28 days in a fused silica-lined canister were 63% and 83% at relative humidity levels of 27% and 53%, respectively. The corresponding values for a polished stainless steel canister were 18% and 93% (Ochiai et al., 2002).

Analysis of low concentrations of butyl acetate in water often requires a preconcentration stage. Senin et al. (1988) reported a method for the analysis of wastewater using a modified aluminosilicate sorbent, zeolite TsVK XI-a, and analysis by gas chromatography with a double flame ionization detector to give a detection limit of 0.1 mg/litre.

Concentrations of organic solvents such as n-butyl acetate have been quantitatively and quasi-continuously analysed in the waste air of a pharmaceutical production facility by means of infrared spectrometry (Düblin & Thöne, 1989).

Several chromatographic methods to determine butyl acetates and butyl alcohols (to which the acetates are rapidly hydrolysed in the blood) have been published, including one proposed by the US EPA (Spingarn et al., 1982; Uehori et al., 1987; Franke et al., 1988; Streete et al., 1992).

No validated methods for biological monitoring of workers exposed to butyl acetates were identified.

All of the butyl acetate isomers have been found to occur naturally in a range of fruits and food products. The n-butyl, isobutyl, and tert-butyl acetates are found in bananas and related fruits (Bisesi, 1994).

n-Butyl acetate has been identified in sunflower (Helianthus annuus) stems (Buchbauer et al., 1993). It is formed during fermentation in yeast and has also been detected in a wide variety of food products, including milk, cheese, beer, rum, brandy, wine, whiskey, cocoa, black tea, coffee, roasted nuts, vinegar, and honey (Maarse & Visscher, 1989). Concentrations were up to 29.5 mg/kg in apples and up to 0.1 mg/kg in grapes, mangoes, melons, and strawberries. n-Butyl acetate has also been detected in apricots and plums (Gomez et al., 1993) and in nectarines (Takeoka et al., 1988). Concentrations in vinegar were up to 166 mg/kg. In drinks, n-butyl acetate was found in apple juice at levels up to 2.2 mg/kg, in cider up to 1.3 mg/kg, in beer up to 0.2 mg/kg, and in weinbrand (a type of brandy) up to 0.4 mg/kg (Maarse & Visscher, 1989).

Isobutyl acetate is found naturally in raspberries, pears, pineapples, and natural cocoa aroma (Opdyke, 1978), black currants, guava, grapes, melons, peaches, strawberries, tomatoes, soya beans, plums, passion fruit, star fruit, and dill herb (Maarse & Visscher, 1989).

sec-Butyl acetate has been found in vinegar at concentrations of 43–67 mg/kg (Maarse & Visscher, 1989).

Butyl acetates may be formed in the atmosphere during the photochemical oxidation of other chemicals. Butyl acetate has been identified as a product of gas-phase reactions of ethyl-n-butyl ether with hydroxyl radicals in the presence of nitric oxide. The molar yield for this reaction was 0.032 ± 0.001 (Johnson & Andino, 2001). tert-Butyl acetate has been identified as a product of gas-phase reactions of di-tert-butyl ether with chlorine atoms and hydroxyl radicals (Langer et al., 1996). The molar tert-butyl acetate yields were 0.85 ± 0.11 and 0.84 ± 0.11 for reactions with chlorine atoms and hydroxyl radicals, respectively. tert-Butyl acetate is a product of ethyl-tert-butyl ether reactions with hydroxyl radicals in the presence of nitrogen oxides (Smith et al., 1992). The molar tert-butyl acetate yield was 0.13 ± 0.001.

Annual global production of butyl acetate was 528 000 tonnes in 1998, whereas annual production of butyl acetate in the USA was reported to be 170 000 tonnes. For regions outside of the USA, annual production was as follows: Japan, 50 000 tonnes; Mexico, 8000 tonnes; South America, 39 000 tonnes; China (Province of Taiwan), 40 000 tonnes; and Western Europe, 221 000 tonnes (CEH, 1999).

In the USA, about 100 000 tonnes of butanol were consumed in 1997 to manufacture butyl acetate (CEH, 1999), which would amount to the manufacture of approximately 150 000 tonnes of n-butyl acetate (assuming 95% theoretical yield for the chemical conversion). The anticipated 1997–2002 US average annual rate of growth of n-butyl acetate production was estimated at 2.2% (CEH, 1999). In Japan, about 38 000 tonnes of butanol were converted to n-butyl acetate in 1997. Using the same yield assumption as above for the chemical conversion, 1997 production in Japan was calculated to be 56 600 tonnes (CEH, 1999).

Global industrial production of isobutyl acetate in 2002 was approximately 74 000 tonnes (CEH, 2003).

Butyl acetates, especially n-butyl acetate and isobutyl acetate, are used as solvents. The Dutch paint industry was reported to have used 1750 tons [sic] of n-butyl acetate and 1275 tonnes of isobutyl acetate in 1979 (Doorgeest et al., 1986). According to data from the Substances in Preparations in Nordic Countries database (SPIN), total uses in Finland, Denmark, and Norway for 2001 were approximately 28 300 tonnes for n-butyl acetate (in 5200 preparations), 1600 tonnes for isobutyl acetate (in 250 preparations), and 30 tonnes for sec-butyl acetate (in 15–20 preparations), respectively. No information was available on tert-butyl acetate in these three countries. n-Butyl acetate is mainly used as a solvent and a thinner in the production of nitrocellulose lacquers in the protective coatings industry. It is also used in the manufacture of high-polish lacquers and varnishes, in a protective low-viscosity vehicle coating used in the motor industry, and in liquid floor wax (Zaleski, 1992). n-Butyl acetate is further used in:

• the cosmetics industry as a solvent in nail polish, base coats, nail polish removers, and other preparations for manicuring (Toy, 1989);
• the food industry as a component in synthetic flavours, as a component used in articles used for food packaging, and also as a diluent for dyes in inks for marking vegetables and fruits (Zaleski, 1992);
• the production of shoe and leather glues, photographic films, plastics, and safety glass (Zaleski, 1992); and
• the pharmaceutical industry as an extractant (Zaleski, 1992).

Both n-butyl and isobutyl acetates are used in perfumery. Isobutyl acetate is also used as a component of hydraulic fluids and as a solvent in manufacturing lacquers and paint removers. sec-Butyl acetate also serves as a solvent for nitrocellulose and nail enamel and in the production of paper coatings.

tert-Butyl acetate is used as a solvent for lacquers (Zaleski, 1992).

Butyl acetate has been identified as a suitable drilling fluid for use during deep ice-coring projects (Gosink et al., 1991).

In Sweden, in 1988, it has been reported that n-butyl acetate was used in 1795 general products and 205 consumer products, totalling approximately 16 000 tonnes; isobutyl acetate was used in 55 general products and 15 consumer products, totalling approximately 45 tonnes; and sec-butyl acetate was used in 1 general product and no consumer products, totalling approximately 200 kg. tert-Butyl acetate was apparently not used in Sweden during this period (Swedish Work Environment Authority, 2001).

n-Butyl acetate may be released to the environment during its use in industrial coatings installations where the plant is not equipped to recover or incinerate vapours. Quantitative data on the amounts released were not available (IUCLID, 2000). n-Butyl acetate is also released to the environment during its use in lacquers, inks, coatings, and adhesives (IUCLID, 2000).

Releases of n-butyl acetate to the atmosphere from industrial plants in the Netherlands were 1170 and 1280 tonnes in 1990 and 1988, respectively (Berdowski & Jonker, 1993). The corresponding figures for isobutyl acetate were 4.2 and 5.6 tonnes per year.

Releases of butyl acetate to surface water from industrial plants in the Netherlands were 0.5 and 2.9 tonnes in 1990 and 1988, respectively (Berdowski & Jonker, 1993).

Butyl acetate used as a drilling fluid is likely to evaporate to the atmosphere, although some may disperse to the water column (Gosink et al., 1991).

There is a paucity of measured volatilization rates for n-butyl acetate, but its Henry’s law constant values (ranging from 2.85 × 10^–2 to 3.25 × 10^–2 kPa·m³/mol) suggest that n-butyl acetate will evaporate from water at a moderate rate (SIDS, 2001). The volatilization half-life from a model river (EPIWIN model version 3.05; Syracuse Research Corp., 2000) 1 m deep, flowing at 1 m/s, with a wind velocity of 3 m/s, was calculated to be 6.1 h; the half-life from a similar river with a depth of 10 m was 7.4 days (IUCLID, 2000). Corresponding values from a model river and lake were 201 min and 5.3 days (SIDS, 2002). Output from the Mackay Fugacity Model Level III (Syracuse Research Corp., 2000) suggested that n-butyl acetate is likely to be distributed to air (93.4%), water (5.78%), soil (0.063%), and sediment (<0.1%) (SIDS, 2002).

For isobutyl acetate, there is also a paucity of measured volatilization rates, but its Henry’s law constant value of 3.53 × 10^–2 kPa·m³/mol suggests that it will evaporate from water at a moderate rate (SIDS, 2003). The volatilization half-lives from a model river and lake (EPIWIN model version 3.05; Syracuse Research Corp., 2000) were 2.9 h and 5.08 days (SIDS, 2003). Output from the Mackay Fugacity Model Level III (Syracuse Research Corp., 2000) suggested that isobutyl acetate is likely to be distributed to air (12.3%), water (42.7%), soil (44.9%), and sediment (0.106%) (SIDS, 2003).

Using the Mackay Fugacity Model Level III, when tert-butyl acetate is discharged to water, 60% evaporates and 13% is removed by chemical reaction; very little is transferred to sediment. When tert-butyl acetate is emitted to air, only 0.25% deposits in water and soil (Webster & Mackay, 1999).

n-Butyl acetate and isobutyl acetate in solution will both undergo hydrolysis reactions to form acetic acid. These reactions follow second-order kinetics and are dependent upon the concentrations of the catalyst ions, hydrogen and hydroxyl. For n-butyl acetate, hydrolysis is more rapid at pH values greater than 5.5 (Johannes et al., 1997). Half-lives for n-butyl acetate calculated at 20 °C ranged from 11.4 days at pH 9 to 114 days at pH 8 to 3.1 years at pH 7 (SIDS, 2002). Half-lives for isobutyl acetate calculated at 20 °C using the HYDROWIN model version 1.67 (US EPA, 2000) ranged from 3.3 years at pH 7 to 122 days at pH 8 (SIDS, 2003).

n-Butyl acetate in the atmosphere will undergo reactions with hydroxyl radicals to form 2-oxobutyl acetate and 3-oxybutyl acetate. Experimentally derived rate constants for this reaction were calculated to be 5.2 ± 0.5 × 10^–12 cm³/molecule per second (Veillerot et al., 1996), 5.71 ± 0.94 × 10^–12 cm³/molecule per second (Williams et al., 1993), and 3.29 ± 0.35 × 10^–12 cm³/molecule per second (Ferrari et al., 1996). Other atmospheric processes, such as direct photolysis, wet deposition, and dry deposition, are not expected to play an important role in the removal of n-butyl acetate from the atmosphere (SIDS, 2001).

tert-Butyl acetate in the atmosphere will undergo reactions with chorine atoms. The rate constant for this reaction was calculated to be 1.6 ± 0.3 × 10^–11 cm³/ molecule per second (Langer et al., 1996). Reactions with hydroxyl radicals gave a rate constant of 4.4 ± 0.4 × 10^–13 cm³/molecule per second. Products formed from reactions of tert-butyl acetate with hydroxyl radicals in the presence of nitric oxide were acetic anhydride and acetone, with molar formation yields of 0.49 ± 0.05 and 0.20 ± 0.02, respectively (Tuazon et al., 1998).

The photochemical removal of isobutyl acetate as mediated by hydroxyl radicals occurs with calculated half-lives of 1.9–2.3 days (SIDS, 2003).

The K_oc for n-butyl acetate was calculated to be 233 (Karickhoff et al., 1979), whereas that for isobutyl acetate was calculated (using the PCKOCWIN model version 1.66; US EPA, 2000) to be 17.5 (SIDS, 2003). Log K_ow values ranging from 1.81 to 1.82 for n-butyl acetate and of 1.78 for isobutyl acetate suggest that both are unlikely to partition from water to soil, sediment, or biota, and they therefore may be leached through soil to groundwater (SIDS, 2001, 2003).

n-Butyl acetate is readily biodegradable. Eighty-three per cent of n-butyl acetate was degraded within 20 days by a non-adapted culture from domestic sewage sludge, while 61% was degraded in seawater. Measured chemical oxygen demand was reported to be 2.32 mg/ mg, with a theoretical oxygen demand of 2.20 mg/mg (Price et al., 1974). Isobutyl acetate is also readily biodegradable. In the same study, 81% of isobutyl acetate was degraded by sewage sludge, and 37% was degraded in seawater within 20 days.

Biodegradation of tert-butyl acetate by US EPA-approved Polyseed was 28% in 28 days. Using acclimated bacteria, biodegradation was 70% or 75% in 28 days (M.I. Banton, personal communication, 1998). Thus, depending on the microorganisms present, tert-butyl acetate is either inherently biodegradable or readily biodegradable.

A stable microbial population consisting of seven strains of bacteria and three strains of yeast, isolated from various samples of soil, water, and activated sludge, was able to completely degrade initial concentrations of up to 10 g/litre of a mixture of butyl acetate and xylene within 96 h (Gardin et al., 1999). The mixture contained 70% xylene (meta- and ortho-isomers) and 30% butyl acetate (isomers not specified). Degradation was higher in a two-phase aqueous:silicone oil-phase system, with a degradation rate of 53 mg/litre per hour for butyl acetate (Gardin et al., 1999).

Five species of fungus were able to use n-butyl acetate vapour as the sole source of carbon and energy. Species used in this test were Cladosporium resinae, Cladosporium sphaerospermum, Exophiala lecanii-corni, Mucor rouxii, and Phanerochaete chrysosporium, and significant growth of each fungus was reported within 30 days at pH 3.5, 5.0, and 6.5 (Qi et al., 2002).

The low log K_ow values suggest that n-butyl acetate, isobutyl acetate, and sec-butyl acetate are unlikely to be bioaccumulated by organisms. A BCF in fish of 14 was calculated for n-butyl acetate using its log K_ow (Staples, 2001), whereas a BCF in fish of 4.7 was calculated for isobutyl acetate using the log K_ow for isobutyl acetate (SIDS, 2003). tert-Butyl acetate has a negligible tendency to bioaccumulate. The BCF is less than 5 (Webster & Mackay, 1999).

In a German field study in selected representative households, low levels of n-butyl acetate were found, at concentrations ranging from not detected (detection limit not given) to 23 µg/m³. The total concentration of volatile organic compounds was 2–3 times higher in winter than in summer (Seifert et al., 1989). n-Butyl acetate was detected at median concentrations of 2–5 µg/m³ in private homes in England (location unspecified) (Crump, 1995), and it was identified in 69% of samples from 26 homes in Finland (location unspecified) (Kostianen, 1995).

In a Swiss study of new and recently renovated buildings, a butyl acetate concentration of 549 µg/m³ was measured. Butyl acetate was found to be off-gassed from a sealing wax on a cork floor (Rothweiler et al., 1992).

Butyl acetate concentrations of 0.1 and 4.8 µg/m³ emanating from US industrial and chemical waste disposal sites have been reported (Pellizzari, 1982).

n-Butyl acetate was present in water samples from seven of the eight small rivers that act as tributaries to Lake Constance (south-west Germany) (Jüttner, 1992). Isobutyl acetate was present in only one tributary. The concentrations were not quantified.

A summary of levels of butyl acetates in workplace air is presented in Table 2. The occurrence of n-butyl acetate particulates in paint spray aerosols has been investigated in six US commercial furniture facilities where sealers and lacquers containing 13–42% (w/w) n-butyl acetate were used. Theoretically, n-butyl acetate in paint particles will vaporize very quickly (e.g., 0.5–1 s for a 20-µm particle). In practice, breathing zone 8-h time-weighted average measurements (24 data sets) showed a mean total (i.e., vapour plus particles) exposure level of 19 mg/m³ (range 5.2–48.3 mg/m³), of which the particle exposure (mean 3.8 mg/m³; range not detected – 11.0 mg/m³) contributed about 20% (Williams, 1995).

Table 2: Occupational air levels (personal air sampling).

^a n-Butyl acetate was found in 2 out of 22 air samples.

^c Data from two subjects.

In Belgium, isobutyl acetate was identified in the air of 5% of operations using printing pastes and inks, 17% using paints and varnishes, 45% of auto repair shops, and about 30% of miscellaneous industries (Veulemans et al., 1987). Isobutyl acetate was detected in shoe and leather factories in Italy at concentrations of 0.2–1.6 mg/m³ (Cresci et al., 1985). A mean concentration of 0.5 mg/m³ was detected in the workplace air of a US company performing spray painting and glueing (Whitehead et al., 1984). Isobutyl acetate was determined in the breathing zone of an automobile paintshop in Spain at concentrations of 37.6, 109.6, and 134.0 mg/m³ (de Medinilla & Espigares, 1988). n-Butyl acetate was detected in the range of 0.02–0.79 mg/m³ during 1 work week in 10 houses above the workrooms of screen printing plants in inner Amsterdam (Verhoeff et al., 1988). A study was conducted among 196 paint workers in two paint manufacturing factories and 25 various kinds of spray painting factories in Tapei, China (Province of Taiwan). Workers were exposed to mixtures of organic solvents, and butyl acetate was among the eight most frequently used. It was detected (73 samples taken during a 24-h period) at a range of 0–200 mg/m³ (Wang & Chen, 1993).

Data are insufficient to estimate human exposure from all routes. JECFA (1998) estimated current levels of intake of n-butyl acetate from use as a flavouring agent in food as 170 μg/person per day in the USA and 1200 μg/person per day in Europe (see section 12). This is likely to be a minor source of overall human exposure.

The most common routes of entry of the butyl acetate isomers into the body are via the lungs and through the skin. Their presence in fruit and a range of other food products makes the oral route also of importance. Although no published quantitative data on absorption were identified, it is expected that the butyl acetate isomers would be absorbed readily by the respiratory tract, the skin, and the gastrointestinal tract.

Human blood/air and rat blood/air partition coefficients for n-butyl acetate were experimentally determined to be 677 and 1160, respectively; those for isobutyl acetate were found to be 578 and 880, respectively (Kaneko et al., 1994). Some rat tissue/blood partition coefficients for these butyl acetates are presented in Table 3.

Table 3: Rat tissue/blood partition coefficients for n-butyl and isobutyl acetate.^a,b

^a From Kaneko et al. (1994).

^b Calculated as (tissue/air) / (blood/air).

n-Butyl acetate, isobutyl acetate, and sec-butyl acetate may be readily hydrolysed to acetic acid and their respective alcohols in the blood, liver, small intestine, and respiratory tract, as has been shown in a number of in vitro experiments using homogenates from liver, small intestinal mucosa, and ethmoturbinates (Longland et al., 1977; Dahl et al., 1987). tert-Butyl acetate is less readily hydrolysed. When added to blood samples from male volunteers or female rats, respective hydrolysis half-lives of n-butyl acetate were 4 and 12 min, while those of tert-butyl acetate were 300 and 270 min (Essig et al., 1989).

The acetic acid is oxidized via the citric acid cycle to carbon dioxide and water. n-Butanol and isobutanol are rapidly metabolized by alcohol dehydrogenase to the respective aldehyde and by aldehyde dehydrogenase to the corresponding acids. These are oxidized further to carbon dioxide. Small amounts of isobutanol may be excreted unchanged or conjugated as a glucuronide (IPCS, 1987).

sec-Butanol is also metabolized by alcohol dehydrogenase, and the metabolite methyl ethyl ketone is excreted in the breath or urine or is further metabolized, producing 3-hydroxy-2-butanone and 2,3-butanediol (IPCS, 1987).

tert-Butanol, however, is a poor substrate for alcohol dehydrogenase and is only slowly metabolized in mammals. It is eliminated in urine as a glucuronide conjugate and as acetone and via the breath as acetone and carbon dioxide (IPCS, 1987).

When a single dose of approximately 30 mg/kg body weight of ¹⁴C-labelled n-butyl acetate (in 0.9% sodium chloride; 0.59–0.67 MBq/animal) was injected intravenously into the tail vein of male Sprague-Dawley rats (n = 32), the isomer was eliminated very rapidly from the blood, with a half-life of 0.4 min. Following dosing, [¹⁴C]n-butyl acetate was detected in brain tissues within the first 2.5 min, reaching a maximum concentration of 3.8 µg equivalents/g tissue after approximately 2 min. Maximum levels of the metabolite [¹⁴C]n-butanol of 52 and 79 µg equivalents/g tissue were found in whole blood and brain, respectively, approximately 2.5 min after dosing. The metabolite was rapidly eliminated from both blood and brain (with a half-life of approximately 1 min); 20 min after dosing, concentrations were below the detection limit (not given). Other metabolites detected in the blood, although to only a minor degree in the brain, included n-butyric acid (with a maximum of 5.7 µg equivalents/g whole blood at 7.4 min, followed by a slow decrease) and polar metabolites (i.e., citric acid cycle intermediates, glucuronide and sulfate conjugates; with a maximum level of 12.2 µg equivalents/g tissue at 4.2 min) (Deisinger & English, 1997).

When nembutal-anaesthetized rats were exposed for 1 h to n-butyl acetate at 33 880 mg/m³ via a tracheal cannula, a nearly constant n-butyl acetate blood level of 140 µmol/litre (16.3 mg/litre) was reached within 1 min. No n-butyl acetate could be detected 1 min after the exposure had finished. Blood levels of n-butanol increased over 40 min of exposure to 480 µmol/litre (35.6 mg/litre). When the exposure was stopped, n-butanol was eliminated from the blood with a half-life of 5 min (Essig et al., 1989).

In a similar experiment, groups of five rats were exposed for 5 h to n-butyl acetate at a concentration of 4840 mg/m³. n-Butyl acetate and n-butanol concentrations in the blood were measured during the first hour at 10-min intervals and during the next 4 h at 15-min intervals. After a steady increase, followed by a slight decrease, the concentration of n-butyl acetate reached a nearly constant level of 24.6 ± 3.8 µmol/litre blood (2.9 ± 0.4 mg/litre) at about 1 h. The concentration of n-butanol followed a similar pattern, reaching a level of 52.4 ± 10.3 µmol/litre (3.9 ± 0.8 mg/litre). When animals were given, after 30 min of exposure, a single intraperitoneal injection of ethanol (790 mg/kg body weight), the amount of n-butanol in the blood was doubled, while mean n-butyl acetate levels were slightly lower (Groth & Freundt, 1991). Alcohol dehydrogenase, involved in the metabolism of n-butanol to its aldehyde, may be inhibited or retarded by ethanol. The increase in n-butanol is thus explained by substrate competition between both alcohols and the alcohol dehydrogenase with ethanol in excess.

Experiments were also performed in rats with the isomer tert-butyl acetate. Inhalation of 22 264 mg/m³ for 2 h resulted in continuously increasing blood levels to approximately 400 µmol/litre (46.5 mg/litre). When exposure ceased, tert-butyl acetate was eliminated in two phases, with half-lives of 5 and 70 min. Blood levels of the metabolite tert-butanol increased continuously throughout the experimental period of 300 min (Essig et al., 1989). When rats were exposed to about 2100 mg/m³, blood levels of both tert-butyl acetate and tert-butanol steadily increased during the 5-h experimental period, with tert-butyl acetate levels generally exceeding those of tert-butanol. At 4 h, these levels became approximately equal; tert-butyl acetate levels reached a plateau value of about 285 µmol/litre (33.1 mg/litre), while tert-butanol continued to increase to approximately 340 µmol/litre (25.2 mg/litre) by the end of the experiment. During exposure to 4356 mg/m³ for 4.25 h, peak concentrations of approximately 450 and 550 µmol/litre (52.3 and 40.8 mg/litre) were measured for tert-butyl acetate and tert-butanol, respectively. Thereafter, tert-butyl acetate levels rapidly declined to approximately 250 µmol/litre (29.0 mg/litre) within 15 min (the end of the experiment), while the tert-butanol level remained constant (Groth & Freundt, 1991).

Rats exposed via inhalation to isobutyl acetate at 9700 mg/m³ in a closed chamber had blood concentrations of isobutanol twice those of isobutyl acetate at both 5 and 10 min (Poet, 2003). From 10 to 25 min into the exposure, the blood isobutanol concentrations were approximately 2- to 2.5-fold higher than the corresponding isobutyl acetate levels.

In vitro experiments have demonstrated that oxidative cytochrome P450-mediated mechanisms may play a role in the cleavage of acetate esters. Using microsomes isolated from phenobarbital-induced rat livers, butyl acetate (at a concentration of 10%, as higher concentrations disrupt the microsomal suspension) bound to cytochrome P450 (type I) stimulated carbon monoxide-inhibitable NADPH oxidation in a way typical for cytochrome P450 substrates. It did not alter cytochrome P450, cytochrome b5, or NADPH-cytochrome c reductase levels (Ivanetich et al., 1978).

For the oxidation of n-butyl acetate by cytochrome P450 2E1, the major ethanol-inducible isoform purified from rabbit liver, a K_M of 1.5 mmol/litre and a V_max of 0.15 nmol aldehyde formed per minute per nanomole P450 were determined (Peng et al., 1995).

Using a reconstituted system containing cytochrome P450 2B4, the major phenobarbital-inducible isoform purified from rabbit liver, sec-butyl acetate was demonstrated to undergo hydroxylation to an unstable hemiketal (2-hydroxy-2-acetoxybutane) followed by a non-hydrolytic cleavage to 2-butanone (methyl ethyl ketone) (Peng et al., 1995).

n-Butyl acetate is probably excreted via exhaled air and urine both as the unchanged compound and as metabolites after transformation in the body. Humans exposed to atmospheres containing n-butyl acetate at a concentration of 200 mg/m³ were reported to excrete 50% of the inhaled compound in the exhaled air (Anonymous, 1992). No data on elimination for the other isomers were identified.

A pharmacokinetic model for n-butyl acetate and its metabolites (butanol, butyraldehyde, and butyrate) has been developed with the objective of formulating a family approach for estimating reference concentrations/doses using internal dose metrics for a series of metabolically related organic chemicals (Barton et al., 2000). This was provisionally parameterized based on limited literature and experimental data. The model consists of submodels for each chemical linked by metabolism and includes compartments for the liver, lung, fat (for the n-butyl acetate component), other tissues, arterial blood, and venous blood. Fat was not included in the models of the metabolites due to their lower lipophilicity. The rate of metabolism was described using a Michaelis-Menten equation (in which metabolism is a function of the maximum metabolic rate for that tissue), the free concentration in tissue, and the concentration at which half-maximal activity occurs. Three routes of administration were included: intravenous injection, oral intubation, and inhalation. The model was implemented for adult rats exposed to n-butyl acetate, and the limited available pharmacokinetic data were used to estimate values for metabolism and clearance parameters. The data allowed development of an initial set of values for the chemical-specific parameters. As an example, using a NOAEC of 2400 mg/m³ for inhalation exposure (6 h/day) to n-butyl acetate identified from the 13-week toxicity study of Bernard & David (1996) (see section 8.4), the model estimated that an equivalent NOAEC for n-butanol would be 2500 mg/m³ (when effects are proportional to blood concentrations of n-butanol or its metabolites), the higher dose reflecting the difference in respiratory tract absorption between the two molecules.

In an unpublished study, the absorption, distribution, metabolism, and excretion of tert-butyl acetate were investigated following inhalation of 480 or 4800 mg/m³. Metabolism of tert-butyl acetate appeared to follow two pathways — one involving hydroxylation to produce 2-hydroxyisopropyl acetate (20% at 480 mg/m³) and the second involving cleavage of the ester linkage to produce tert-butanol and acetic acid (80% at 480 mg/m³). At 4800 mg/m³, 69% of the inhaled dose was eliminated in the urine and 27% in expired air. At 480 mg/m³, 89% was in the urine and 5% in air, suggesting that metabolism of tert-butyl acetate may be somewhat saturated at 4800 mg/m³ (Girkin & Kirkpatrick, 2000).

8.1.1 n-Butyl acetate

Data on the acute toxicity of n-butyl acetate by the inhalation route are summarized in Table 4. The results from studies in rats show that exposure to nearly saturated atmospheres generated by evaporation did not result in death. The data from atmospheres/aerosols generated by atomizers are highly inconsistent, with LC₅₀ values ranging from 740 mg/m³ to above 45 000 mg/m³. Following the report of an LC₅₀ of 740 mg/m³ for aerosolized n-butyl acetate (Debets, 1986), further studies were conducted at three different laboratories in an attempt to replicate the data, to differentiate between data from vapours and aerosols, and to investigate the role of small particles and of relative humidity (unpublished studies reviewed in Norris et al., 1997). These inconsistencies occurred not only between laboratories, but also within the same laboratory. Using identical inhalation equipment and aerosol generation procedures, one laboratory observed no mortality at concentrations up to approximately 21 395 mg/m³. In a second laboratory, LC₅₀s of approximately 1900 mg/m³ and 5300 mg/m³ were determined, while no deaths occurred in a third experiment with exposures up to 45 000 mg/m³. In the third laboratory, findings not observed in the other two laboratories included low chamber relative humidity, brief times to death (all dead within 24 h post-exposure, 7/10 animals in the highest concentration group died in the last 2 h of exposure versus mortality 1–4 days post-exposure in the other studies), and the histological finding of vesicular emphysema, suggesting that there might have been methodological problems in this study. The explanation for the inconsistent results from exposure to aerosolized n-butyl acetate is not known (Norris et al., 1997).

Table 4: Effects on experimental animals due to acute inhalation exposure to n-butyl acetate.

Clinical signs observed in rats during acute inhalation exposures (from atomizers) ranged from eye irritation (periocular wetness, blepharospasms) to nervous system effects (hypoactivity, ataxia, forced/shallow breathing, narcosis). At gross necropsy of the deceased animals, discoloration of the lungs and fluid in the thoracic cavity and trachea were observed. Microscopic examination of the lungs from some animals revealed congestion, alveolar haemorrhage, sloughing of bronchiolar mucosa, necrosis of alveolar epithelial cells, and oedema (Nachreiner & Dodd, 1987; Nachreiner, 1993). Discoloration of the lungs was also observed in rats surviving a 4-h exposure to approximately 23 000 or 43 000 mg/m³. Clinical signs (narcosis, incoordination, perioral wetness) were seen only at the higher level on the exposure day; no clinical signs were observed during the 14-day post-exposure period. Exposure to concentrations of 3900 mg/m³ and above caused blepharospasms (Nachreiner, 1994).

There were apparently no deaths when four groups of 20 rats (10 per sex) were exposed for 6 h to 0, 7200, 14 000, or 29 000 mg/m³ of n-butyl acetate vapours generated by evaporation (and not present in aerosol form). Body weight decreases, over 14 days following exposure, between treated and control animals did not exceed 10%, but were statistically significant for the male animals of the low-dose (on post-exposure day 7) and high-dose (on post-exposure days 7 and 14) groups (Bernard & David, 1994).

Data on the acute toxicity of n-butyl acetate by other routes are summarized in Table 5. They indicate that n-butyl acetate has low toxicity by the oral and dermal routes.

Table 5: Effects on experimental animals after single oral or dermal exposure to n-butyl acetate.

^a Except where otherwise noted.

^b ND₅₀ = the quantity that produced stupor and loss of voluntary movements in half of the animals.

8.1.2 Isobutyl acetate

Data on the acute toxicity of isobutyl acetate are presented in Table 6. They indicate that isobutyl acetate has low toxicity via the inhalation, oral, and dermal routes.

Table 6: Effects on experimental animals after acute exposure to isobutyl acetate.

^a Units are mg/m³ for inhalation routes, g/kg body weight for oral and dermal routes.

^b ND₅₀ = the dose that produced stupor and loss of voluntary movements in half of the animals.

8.1.3 sec-Butyl acetate

An unpublished report (Roudabush, 1970) states that all rats survived exposure to sec-butyl acetate for 6 h at approximately 17 000 mg/m³, while all rats died when exposed for 4 h to 116 000 mg/m³. An oral LD₅₀ of 3200–6400 mg/kg body weight was also reported for rats (no further details available).

8.1.4 tert-Butyl acetate

For tert-butyl acetate, a 4-h LC₅₀ of 13 300 mg/m³ has been determined for Sprague-Dawley rats by exposing groups of five per sex to aerosol concentrations of 5000, 10 000, 15 000, or 30 000 mg/m³ (particle size and distribution not given). Symptoms observed included inactivity and sedation, hyperactivity comparable to the excitation state of anaesthesia, coma, and death. Although the onset of the effects was much shorter at the higher concentrations, the time course of clinical signs during the exposure period was generally similar at all concentrations. Postmortem examination showed some evidence of pulmonary congestion and haemorrhage only (observation time 14 days) (Kay, 1953). In another study, all rats (Harlan Sprague-Dawley; five per sex) survived nose-only exposure to a mean vapour concentration of 2230 mg/m³ for 4 h. Apart from slight weight loss between days 0 and 7 in one female and red penile discharge in one male animal, no abnormalities were observed in body weight, clinical, or gross necropsy observations (observation time 14 days) (Bennick, 1997). A 6-h LC₅₀ of 20 000 mg/m³ has been determined for Sprague-Dawley rats by exposing groups of five per sex to vapour concentrations of 9000, 17 000, or 24 000 mg/m³. Symptoms observed included exaggerated breathing immediately post-exposure, periodic shaking of the head and thorax, immobility and lethargy, cold to touch, unconsciousness, and death. Postmortem examination showed evidence of pulmonary congestion in decedents. No compound-related pathology was seen in survivors (observation time 14 days) (Kenney, 1999).

An oral LD₅₀ of 3.8 ml/kg body weight (approximately 3420 mg/kg body weight) was estimated in rats (Sprague-Dawley; five per sex per group) by using eight dose groups and a dose range of 1.0–12.0 ml/kg body weight. At 1.0 ml/kg body weight, only a slight restlessness was observed. At doses of 2.0 ml/kg body weight and above, initial restlessness was followed by ataxia, coma, and death. With increasing doses, the severity and incidence of effects increased and time of onset of effects decreased. No tissue or organ changes were observed in the dead animals at postmortem examination (observation time 14 days) (Kay, 1953). An oral LD₅₀ of 4.5 g/kg body weight (males: 4.1 g/kg body weight; females: 4.75 g/kg body weight) was determined in another study in which Wistar rats (five per sex per group) were given 2.0, 5.0, or 7.0 g/kg body weight. Clinical signs observed included ataxia, flaccid muscle tone, lethargy, dyspnoea, loss of righting reflex, prostration, piloerection, tremors, and coma. Necropsy findings in the surviving animals were normal; in those animals dying as a result of treatment, there were abnormalities in various organs as well as wetness and red and brown staining of the nose and mouth area (DeGeorge, 1997d).

No mortality or effects on body weight were found in New Zealand White rabbits (five per sex per group) following 24-h covered contact between tert-butyl acetate at 2000 mg/kg body weight and the clipped intact dorsal skin. There were instances of diarrhoea in 3 of 10 animals during the first week following exposure. Apart from kidney abnormalities in one female animal, no abnormalities were observed on postmortem macroscopic examination (DeGeorge, 1997c). No overt toxicity was observed following 24-h covered application of single doses ranging from 2.0 to 23.0 ml/kg body weight (approximately 1800–20 700 mg/kg body weight) to the clipped skin of New Zealand White rabbits (two per sex per group) (observation time 14 days) (Kay, 1953).

8.2.1 n-Butyl acetate

Following 24-h application of 0.01 ml of the neat material to the clipped skin of five albino rabbits, n-butyl acetate was at most only slightly irritating (Smyth et al., 1954). When 0.5 ml was applied to the clipped intact dorsal skin of New Zealand White rabbits (n = 5) under gauze patches and loosely covered with impervious sheeting for 4 h, no irritation was observed over an observation period of 14 days. Severe irritation occurred, however, if the occlusion period was 24 h (Bushy Run Research Center, 1987; Myers & Tyler, 1992).

Slight irritation was observed when 0.1 ml of n-butyl acetate (99% purity) was instilled into the conjunctival sac of four rabbits for 24 h. A maximum Draize score of 7.5 (out of a possible total of 110) was recorded; scores at 48 h, 72 h, and 7 days were 2.0, 2.0, and 0.5, respectively (ECETOC, 1992). In a similar study, iritis and minor to moderate conjunctivitis (both of which had healed within 48 h), but no corneal damage, were observed when 0.1 ml was instilled into the eyes of six rabbits. A maximum Draize score of 14.7/110 (occurring at 4 h) was recorded (Bushy Run Research Center, 1987; Myers & Tyler, 1992). Following instillation of 100%, 30%, 10%, and 3% n-butyl acetate into the conjunctival sac of rabbits for 24 h, Kennah et al. (1989) reported Draize scores of 8, 11, 19, and 2, respectively (no further details given).

However, in an early study, n-butyl acetate was rated as a severe irritant after a 5-µl volume was instilled into the eyes of rabbits (Smyth et al., 1954). Eye irritation was observed in guinea-pigs exposed for 5 min to atmospheres containing n-butyl acetate at approximately 16 000 mg/m³ (Sayers et al., 1936). Exposure to 2420 mg/m³ for 10 (guinea-pigs) or 20 (rabbits) days or to 4840 mg/m³ for 4 days (guinea-pigs, rabbits) did not result in corneal or conjunctival injury or in changes in corneal sensation (Anonymous, 1992).

Irritation of the respiratory tract has been investigated by determining the concentration associated with a 50% decrease in the respiratory rate (RD₅₀). Using Swiss OF1 mice (n = probably 10), the RD₅₀ for n-butyl acetate was approximately 3470 mg/m³ (Muller & Greff, 1984; Bos et al., 1992). An RD₅₀ of approximately 8340 mg/m³ was determined in another study using male BALB/c mice (n = 8–10) (Korsak & Rydzynski, 1994). In a 13-week inhalation study in which rats were exposed 6 h/day, 5 days/week, olfactory epithelial necrosis was reported, of minimal to mild severity at 7260 mg/m³ and of mild to moderate severity at 14 520 mg/m³. No such lesions were observed after an exposure to 2662 mg/m³ (Anonymous, 1996; Shulman, 1996).

n-Butyl acetate showed no sensitization potential when tested in a maximization test using guinea-pigs or in a mouse ear swelling test (Gad et al., 1986). In the maximization test, 15 Hartley strain guinea-pigs were each given intradermal injections of n-butyl acetate together with an adjuvant, followed 7 days later with a 48-h covered patch. A challenge patch (24-h covered contact) was applied 7 days after this induction regimen. In the mouse study, groups of 10–15 animals were given intradermal injections of an adjuvant and repeated skin applications of n-butyl acetate. After a 7-day non-treatment period, a topical application of n-butyl acetate was made to one ear, the other acting as control. Ear thickness was assessed 24 h and 48 h following this challenge.

8.2.2 Isobutyl acetate

Isobutyl acetate has been tested for skin and eye irritation, although not using protocols that would meet modern regulatory guidelines. The isomer caused no irritation to rabbit skin (scoring grade 1 on a scale of 1–10) following uncovered application of 0.01 ml of an undiluted sample for 24 h (Smyth et al., 1962). Data from an unpublished report submitted to the US Research Institute for Fragrance Materials suggested that the neat material was moderately irritating when applied under occlusion for 24 h to the intact or abraded skin of rabbits (Opdyke, 1978).

The neat material (0.5 ml) was reported to cause a moderate inflammation in the eyes of rabbits (scoring grade 2 on a scale of 1–10) (Smyth et al., 1962).

Irritation to the respiratory tract has been investigated in mice. The RD₅₀ was 3890 mg/m³ (Muller & Greff, 1984; Bos et al., 1992).

Isobutyl acetate was apparently not a skin sensitizer in guinea-pigs (no further details on this unpublished study available) (Huels AG, 1988a).

8.2.3 sec-Butyl acetate

No data were identified on the irritation or sensitization potential of sec-butyl acetate.

8.2.4 tert-Butyl acetate

The primary skin irritation potential of tert-butyl acetate has been tested by applying 0.5 ml of the neat material to the clipped intact dorsal skin of New Zealand White rabbits (three per sex) under gauze patches, semi-occlusively wrapped with plastic, for 4 h. The wrappings were then removed, the residual test compound was washed off with distilled water, and the skin was scored for irritation at 30–60 min and at 24, 48, and 72 h following removal of the patch. Very slight, barely perceptible erythema (scoring 1 on a scale of 0–4) was observed in 6/6, 4/6, 0/6, and 0/6 animals at 30–60 min and 24, 48, and 72 h, respectively. Oedema was absent at all observation intervals. No ulceration, necrosis, or any other evidence of tissue destruction was observed (DeGeorge, 1997a).

No dermal responses (erythema or oedema) were observed during a 14-day observation period following 24-h covered contact between tert-butyl acetate and the clipped intact dorsal skin of New Zealand White rabbits (DeGeorge, 1997c). Erythema, which had cleared within 48 h, was the only effect reported following 24-h covered contact with the clipped skin of New Zealand White rabbits (two per sex per group) (observation time 14 days) (Kay, 1953).

The potential of tert-butyl acetate to cause eye irritation was tested by instilling 0.1 ml into the conjunctival sac of one eye of male New Zealand White rabbits (n = 6). Treatment induced corneal opacity in 1/6 (which had cleared by day 2), iritis in 3/6 (cleared by day 2), and conjunctival irritation in 6/6 animals (cleared by day 3) and resulted in mean Draize scores of 14.5, 6.8, 2.0, 0, and 0 (out of a possible total of 110) at observation times of 1, 24, 48, and 72 h and 7 days, respectively (DeGeorge, 1997b). In another study, instillation of 0.1 ml into the conjunctival sac of five New Zealand White rabbits caused minimal conjunctival irritation, which lasted for 96 h. Mean Draize scores were 4.8, 3.6, 2.0, 2.0, 1.6, and 0 at observation times of 1, 24, 48, 72, and 96 h and 7 days, respectively (Kay, 1953).

8.3.1 n-Butyl acetate

In an unpublished study conducted to select exposure concentrations for a subsequent 13-week study (see section 8.4 below), male and female Sprague-Dawley rats were exposed to n-butyl acetate vapour at approximately 0, 3630, 7260, or 14 520 mg/m³, 6 h/day, 5 days/week, for 2 weeks. Each exposure group consisted of five male and five female ad libitum-fed animals and five feed-restricted male animals. There were treatment-related reductions in activity levels (hypoactivity; slower response to tapping on the chamber wall). In the 3630 mg/m³ group, these reductions were of "minimal to minor" severity early in the exposure and absent by the end of the experiment. At 7260 mg/m³, the severity of the effect decreased from "minor" to "minimal" over the course of the exposure, while in the 14 520 mg/m³ exposed group, it remained "minor" throughout the experimental period. Other occasional signs noted were sialorrhoea (excessive saliva and drooling) in 4/15 and 8/15 animals at 7260 and 14 520 mg/m³, respectively, and red sialorrhoea, porphyrin tears and nasal discharge, brown discoloured facial hair, and unkempt hair coat in individual animals of the 14 520 mg/m³ group. There was no apparent difference in these clinical signs between ad libitum-fed and feed-restricted animals. Apart from two animals of the feed-restricted 14 520 mg/m³ group, animals in all treated groups were normal following the exposure. Some transient effects on mean body weights were observed (decreases in female animals at 7260 mg/m³ and in male and female animals at 14 520 mg/m³), but a statistically significant decrease in mean terminal body weight and in mean body weight gain were observed only in the male animals of the feed-restricted 14 520 mg/m³ exposed group. There were no effects on absolute or relative lung, kidney, or liver weights or histological changes (Bernard & David, 1995). (The extent of microscopic examination was not given in the source document.)

No effects on blood counts, urine examinations, or necropsy data (not further defined) were reported in (unspecified numbers of) guinea-pigs exposed to 4840 mg/m³, 4 h/day, for 28 days (Anonymous, 1992).

When (an unspecified number of) cats were exposed to atmospheres containing approximately 20 000 mg/m³, 6 h/day for 6 days, weakness, weight loss, and minor changes in blood values were reported. At approximately 15 000 mg/m³, changes in blood cell morphology were observed, and at 7600 mg/m³, there was increased salivation (Anonymous, 1992).

8.3.2 Isobutyl acetate and sec-butyl acetate

No relevant data were identified on the toxicity of isobutyl acetate and sec-butyl acetate following short-term exposure.

8.3.3 tert-Butyl acetate

In an unpublished study, groups of five male and five female ad libitum-fed Sprague-Dawley rats were exposed to tert-butyl acetate vapour at approximately 0, 580, 2100, or 7900 mg/m³, 6 h/day, 5 days/week, for 2 weeks. No treatment-related clinical signs of toxicity or effects on mean body weight, food consumption, or water consumption were observed in any group. Liver weights were increased in male rats exposed at 7900 mg/m³. Centrilobular hepatocyte hypertrophy was seen in all males at 7900 mg/m³ and in one of five males at 2100 mg/m³. An increased degree of cortical tubules with hyaline droplets was reported in all male groups treated with tert-butyl acetate (Kenney, 2000). (All gross lesions, liver, kidney, nasal turbinates, larynx, and lung were examined microscopically.)

8.4.1 n-Butyl acetate

In an inhalation study with exposure for 13–14 weeks, conducted in parallel with a subchronic neurotoxicity study (see section 8.8 below), groups of 15 male and 15 female Sprague-Dawley rats were exposed to target vapour concentrations of approximately 0, 2400, 7200, or 14 000 mg/m³, 6 h/day, 5 days/week. On day 30, five animals per sex per group were killed for clinical pathology. There was no compound-related mortality in any of the groups. In the 14 000 mg/m³ group, all animals showed slightly reduced activity (defined as less movement, decreased alertness, and slower response to tapping on the chamber wall in comparison with control animals). Occasionally, signs of sialorrhoea and red discoloration of the chin hair were observed. Mean body weights and food intake were generally lower than those of the control animals throughout the study. At the end of the study, weight gains for males and females were lower than those of controls by 38% and 22%, respectively. There were no treatment-related ophthalmological changes or adverse effects on haematology or clinical chemistry parameters. Organ weight changes included decreased absolute liver and kidney weights, decreased absolute and relative spleen weights (males), and increased relative adrenal and lung weights (males). There were also decreased absolute (males) and relative brain weights and increased relative testes weight. On gross and microscopic examination, lesions found were limited to the stomach (minimal haemorrhage in the glandular gastric mucosa in 2/10 females; minimal white discoloration in the non-glandular gastric mucosa in 1/10 females; minimal to mild inflammatory and degenerative lesions of stomach mucosa in 3/10 females) and the nasal passages (olfactory epithelial necrosis of mild to moderate severity in all males and females). At 7200 mg/m³, all animals exhibited slightly reduced activity. Mean body weights were lower at week 6 onwards for males and at week 2 onwards for females. Overall, weight gains were approximately 20–30% lower than those for controls. Food intake was generally lower throughout the study. There were no effects on ophthalmology, haematology, or clinical chemistry parameters. Organ weight changes observed included decreased absolute spleen, liver, and (in females) kidney weights and (in females) increased relative adrenal and brain weights. Males also had an increased relative testes weight. On microscopic examination, histological lesions in the nasal passages consisting of olfactory epithelial necrosis, of minimal to mild extent, in 4 of 10 male animals and in 3 of 10 female animals were observed. No treatment-related effects were observed in the 2400 mg/m³ exposure group. Although numbers of epididymal sperm for all treated groups were lower than controls, the changes were not statistically significant, testicular sperm counts were unchanged, and no dose–response was observed. A NOAEC was therefore considered to be 2400 mg/m³ (Bernard & David, 1996; David et al., 2001).

8.4.2 Isobutyl acetate

No data were identified on the toxicity of isobutyl acetate following medium-term exposure. Data on isobutanol are included here, as these are considered relevant given the rapid hydrolysis of isobutyl acetate to isobutanol.

As part of an evaluation of the potential neurotoxicity of isobutanol (see section 8.8 below), groups of at least 10 male and 10 female Sprague-Dawley rats were exposed 6 h/day, 5 days/week, for up to 14 weeks to isobutanol vapour concentrations of approximately 0, 770, 3100, or 7700 mg/m³. There was a slight (but statistically significant) increase in red blood cell counts, haematocrit, and haemoglobin parameters in females exposed to 7700 mg/m³ when compared with controls. There were no changes in ophthalmology, serum chemistry, organ weights, or gross and microscopic pathology that were attributed to the isobutanol exposure (Li et al., 1999). A NOAEC for isobutanol was 3100 mg/m³.

In an unpublished study, groups of 30 male and 30 female rats were given isobutanol at doses of 0, 100, 316, or 1000 mg/kg body weight per day by gavage for up to 13 weeks. Hypoactivity, ataxia, salivation, laboured respiration, rales, prostration, hypothermia, and emaciation were reported in animals administered the top dose. Hypoactivity and ataxia were the most common clinical signs, and these had largely resolved after 4 weeks of exposure. No clinical signs were seen at 100 or 316 mg/kg body weight per day, and there were apparently no treatment-related effects on organ weights, gross pathology, or histopathology observed (TRL, 1987). The NOAEL was 316 mg of isobutanol per kg body weight per day.

8.4.3 sec-Butyl acetate and tert-butyl acetate

No relevant data were found for sec- or tert-butyl acetate.

No data were found on the long-term toxicity or carcinogenicity of butyl acetates.

No adequate studies on n-butanol, sec-butanol, or isobutanol (to which the respective butyl acetates are readily metabolized) were identified. However, for tert-butanol, long-term carcinogenicity studies have been conducted by the US NTP in which groups of 60 male and 60 female F344/N rats and B6C3F1 mice were given drinking-water containing the alcohol for up to 2 years. For rats, average daily doses were approximately 0, 90, 200, or 420 mg/kg body weight for males and 0, 180, 330, or 650 mg/kg body weight for females. Average daily doses in mice were 0, 540, 1040, or 2070 mg/kg body weight in males and 0, 510, 1020, or 2110 mg/kg body weight in females. In these studies, there was some evidence of carcinogenic activity reported for male rats given drinking-water providing up to approximately 420 mg/kg body weight per day, based on increased incidences of renal tubule adenomas and carcinomas (combined). In a standard evaluation at the end of the study, incidences of combined adenomas and carcinomas were 1/50, 3/50, 4/50, and 3/50. An extended evaluation of the kidney identified additional adenomas and carcinonomas. Incidences of adenomas and carcinomas combined f