2-ETHYL-1-HEXANOL First draft prepared by Dr K. Ekelman, Additives Evaluation Branch Division of Health Effects Evaluation Center for Food Safety and Applied Nutrition Food and Drug Administration, Washington, DC, USA 1. EXPLANATION 2-Ethyl-1-hexanol (EH; also known as 2-ethylhexyl alcohol and 2-ethylhexanol) has not been reviewed previously by JECFA. EH is a colorless liquid with a mild floral odor (Furia and Bellanca, 1975) that occurs naturally in food. EH is prepared by petrochemical synthesis and is used as a flavoring ingredient in food. Total annual consumption of EH in the United States from its natural occurrence in food is reported to be 120 kg (Stofberg and Grundschober, 1987); total annual production for use as a flavor additive is estimated to be 209 kg (NRC, 1989). Estimated intake in the United States from the use of EH as a flavoring ingredient is approximately 0.65 µg/kg bw/day (FEMA, 1993). 2. BIOLOGICAL DATA 2.1 Biochemical aspects 2.1.1 Absorption, distribution, and excretion Two adult male CD-strain rats (300 g) were gavaged with radiolabeled 2-ethyl-1-14C-hexanol (14C-EH; 1 µCi; 8.8 µg) in cottonseed oil. Two others were given the same amount of 14C-EH and cottonseed oil, but also were given 0.1 ml (0.64 mmol) of unlabeled EH. Following administration of the test substance, rats were housed in metabolism cages with ad libitum access to feed and water; expired CO2, urine, and faeces were collected every hour for 28 hrs. Most (99.8%) of the orally administered radioactivity was accounted for by radioactivity in expired CO2, urine, faeces, an ethanol wash of the metabolism cage at the end of the experiment, heart, brain, liver, kidneys, and "residual carcass." EH was efficiently absorbed following oral administration and rapidly excreted in respired CO2 (6-7%), urine (80-82%), and faeces (8-9%); elimination was essentially complete by 28 hrs. The major urinary metabolite of EH in the rat was shown to be 2-ethylhexanoic acid through acid extraction of urine. This metabolite can undergo partial ß-oxidation and decarboxylation to produce 14CO2 and 2- and 4-heptanone (in the urine). Other urinary metabolites of EH were identified as 2-ethyl-5-hydroxyhexanoic acid, 2-ethyl-5-ketohexanoic acid, and 2-ethyl-1,6-hexanedioic acid. Approximately 3% of the parent compound was excreted unchanged. Rats and other mammals hydrolyze orally ingested DEHP (di-[2-ethylhexyl]phthalate, a plasticizer in food-contact materials) to EH and MEHP (mono-[2-ethylhexyl]phthalate) prior to absorption of MEHP by the intestine (Albro, 1975). An in vitro dermal absorption study of EH and seven other compounds was conducted with full thickness rat skin and human stratum corneum. The ratio of the rate of absorption of EH through rat and human skin (rat/human) was reported to be 5.8, indicating that rat skin is more permeable to EH than is human skin. Damage to skin by dermal application of EH was defined as the ratio of the permeability constant for 3H2O after contact with EH to the permeability constant for 3H20 before application. Ratios for human skin (1.5±0.4 and 3.7±2.1) and rat skin (31.9±5.1) indicated that dermal application of EH damages rat skin more than human skin (Barber et al., 1992). Excretion balance studies were conducted on female Fischer 344 rats (4 animals/group) following acute oral doses of 50 or 500 mg/kg bw 14C-EH and repeated oral doses of 50 mg/kg bw/day 14C-EH for 14 days; results of acute gavage doses of 500 mg/kg bw 14C-EH administered neat and as aqueous suspensions containing 5 mg Polyoxyl 35 castor oil/100 ml were compared. Dermal exposures to 14C-EH (1 g/kg bw applied dose) for 6 hours and i.v. exposures to 1 mg/kg bw 14C-EH were also studied. Acute oral doses of 50 or 500 mg/kg bw and repeated oral doses of 50 mg/kg bw/day showed similar excretion balance profiles of 14C, with some evidence of metabolic saturation at the high dose. No evidence of metabolic induction was reported following repeated dosing. All oral doses were rapidly eliminated during the first 24 hours after dosing, predominantly in the urine. Approximately 5% of the dermal dose was absorbed. A majority of the oral and dermal doses were eliminated as glucuronides of oxidized metabolites of EH, principally glucuronides of 2-ethyladipic acid, 2-ethylhexanoic acid, 5-hydroxy-2-ethylhexanoic acid, and 6-hydroxy-2- ethylhexanoic acid. Only trace amounts of unchanged EH were eliminated in the urine. Bioavailability of EH orally administered with the gavage vehicle was slightly greater than bioavailability of EH administered alone (Deisinger et al., 1992). 2.1.2 Biotransformation Knaak and coworkers (1966) studied the metabolism of 8 mg 14C-EH in rats and 200 mg 14C-EH in rabbits following i.p. injection. The major urinary metabolite in rats was 2-ethylhexanoyl glucuronide; EH and 2-ethyl hexanoic acid were also identified in rat urine. In rabbits, the major urinary metabolite also was 2-ethylhexanoyl glucuronide; 2-ethyl-2,3-dihydroxyhexanoic acid and EH were also identified in rabbit urine (Knaak et al., 1966). The metabolism, distribution and elimination of 14C-DEHP following oral administration to male and female B6C3F1 mice, Fischer 344 rats, and Cynomolgus monkeys was investigated. Orally administered DEHP is rapidly hydrolyzed to the monoester (MEHP), then the alcohol and acid. The GI tract had appreciable amounts of EH, and absorbed 14-C appeared to be primarily EH. The alcohol was oxidized by ß-oxidation, omega-, and omega-1-oxidation generating several major and minor products including acids, ketones, ketoacids, hydroxy acids, and diacids (such as ethylhexanoic acid [EHA], ethylhexanedioic acid [DiEHA], and 5-hydroxyethylhexanoic acid [5-OH EHA]). The three species were able to form glucuronic acid conjugates of the alcohol oxidation products, but no sulfites were detected. However, differences between species were noted: metabolism appeared to be less extensive in the monkey (which excreted largely MEHP and EH as their glucuronides) than in rodents (which excreted largely products of faster oxidation, primarily EHA, 5-OH EHA, and diEHA) (Midwest Research Institute, 1984). 2.1.3 Effects on enzymes and other biochemical parameters Gavage administration of 1 mmol/kg bw/day EH (approximately 130 mg/kg bw/day) to five male Wistar rats for 14 days was not associated with liver peroxisome proliferation (Rhodes et al., 1984). However, in another study, administration of 2% EH in the diet (approximately 1 000 mg/kg bw/day) to five male Fischer 344 rats for three weeks was reported to cause peroxisome proliferation and significant increases in the activities of liver catalase and carnitine acetyltransferase (Moody and Reddy, 1978). When gavage doses of 0, 100, 320, or 950 mg/kg bw/day EH were administered to male and female Fischer 344 rats (5/sex/group) for 21 days, significant hepatomegaly at 950 mg/kg bw/day, significant increases in cyanide-insensitive palmitoyl CoA oxidation (a marker for peroxisome proliferation) in males (dose-related, 320 and 950 mg/kg bw/day) and females (950 mg/kg bw/day), and significant increases in lauric acid hydroxylase activity in males and females at 950 mg/kg bw/day were shown. As well, electron microscopy showed only a slight increase in the number of peroxisomes in hepatocytes of high-dose rats (Hodgson, 1987). Groups of five male and five female Alderley Park rats and mice were gavaged with 0, 140, 350, 700, 1050, or 1750 mg/kg bw/day EH for 14 days. Rats in the high-dose group exhibited toxic effects (not specified) and died or were killed during the course of the study. Dose-related increases in relative liver weights of rats and mice were observed; the increases were statistically significant in rats at 700 and 1050 mg/kg bw/day, in male mice at 700, 1050, and 1750 mg/kg bw/day, and in female mice at 1750 mg/kg bw/day. EH administration resulted in a nearly linear dose-related induction of peroxisomal ß-oxidation (measured as palmitoyl CoA oxidation activities) in both rats and mice, although the dose(s) at which this effect was statistically significant were not stated (Keith et al., 1992). Activity of succinic dehydrogenase was increased and activity of lactic dehydrogenase was decreased after 12 daily dermal applications of 2 ml/kg bw undiluted EH to the shaved skin of the rat. In addition, EH-treated rats had significantly lower body weights than control rats 17 days after dermal application of EH was terminated (Schmidt et al. 1973). Microsomal P-450 content increased and glucose-6-phosphatase activity decreased in rat liver microsomal pellets following oral administration of EH to the intact animal. Administration of EH increased alcohol dehydrogenase activity demonstrated histochemically in the centrilobular area of the liver, the number of microbodies, the dilatation of the smooth endoplasmic reticulum, and the number of microperoxisomes in the hepatocytes of rats (Lake et al., 1975). Concentrations of EH ranging from 2.5-15 mM significantly inhibited the activities of rat liver aminopyrine N-demethylase (approximately 60% inhibition at 15 mM EH) and aniline hydroxylase (approximately 50% inhibition at 15 mM EH) in vitro (Seth, 1982). Rhodes and coworkers (1984) reported that 0.1 or 0.5 mM EH did not induce palmitoyl CoA oxidase activity (a marker for peroxisome proliferation) in rat hepatocytes in vitro (Rhodes et al., 1984). The activity of carnitine acetyltransferase (a peroxisomal enzyme) in rat liver cells in vitro was significantly induced (approximately 9X level in untreated cultures) by 1mM EH but not by 0.2 mM EH (Gray et al., (1982). In an in vitro test system using viable plugs from periportal or pericentral regions of rat liver, Liang and coworkers (1991) demonstrated that incubation of these plugs with EH (0.1 to 3 mM) decreased urea synthesis in a dose-related manner (up to 80% inhibition at 800 µM 02) and caused extensive cell damage (assessed by lactate dehydrogenase leakage) Liang et al., 1991). 2.2 Toxicological Studies 2.2.1 Acute toxicity studies Results of acute toxicity studies with EH are summarized in Table 1. Rabbits given 0, 0.2, 0.4, 0.6, 0.8, 1.6 or 3.2 x 10-5 mol/kg EH (0.26 to 4.16 mg/kg bw i.v.) had dose-related increases in heart rate and frequency of respiration. However, when EH was administered to dogs (doses reported as 4.05 and 8.10 mol/kg i.v.), no compound- related hypotensive effects were seen. Finally, when rabbits and rats were given acute iv doses of EH (doses not provided), direct toxic damage to the heart and smooth muscle elements of the blood vessels was observed (Hollenbach et al., 1972). 2.2.2 Short-term toxicity studies 2.2.2.1 Mice Doses of 0, 100, 330, 1 000, or 1 500 mg 2-ethyl-1-hexanol/kg bw/day were administered by gavage for 11 days (9 administrations) to groups of 10 male and 10 female C3B6F1 mice. Effects clearly related to administration of the test substance occurred in male and female mice receiving 330, 1 000, or 1 500 mg/kg bw/day. One male mouse receiving 330 mg/kg bw/day for 11 days showed ataxic gait and piloerection following the administration of the 8th dose, but these symptoms were reported to have disappeared by the next day. Two male and two female mice receiving 330 mg/kg bw/day were reported to have acanthosis in the mucous membrane of the forestomach that was usually associated with hyperkeratosis and was once associated with focal inflammatory oedema in the submucosa. Table 1: Summary of Acute Toxicity Studies with EH Species Route LD50 (mg/kg bw) Reference Rat Oral 2053 (2.46 mL/kg) Smyth et al., 1969 Rat Oral 37301 Scala and Burtis, 1973 Rat Oral 3 250 Albro, 1975 Rat Oral 3200 - 6400 Treon, 1963 Rat Oral 3200 NIOSH, 1976 Rat I.P. 650 Treon, 1963 Mice I.P. 780 Treon, 1963 Rat I.V. 1670 Mashkina, 1966 Mice I.V. 1670 Mashkina, 1966 Rabbit Dermal 1986 (2.38 mL/kg)2 Smyth et al., 1969 Rabbit Dermal >26003 Scala and Burtis, 1973 Guinea pig Dermal >8300 (>10 mL/kg) Treon, 1963 LC50 Mice Inhal. >227 ppm (6 hr)4 Scala and Burtis, 1973 Table 1 (contd). Species Route LD50 (mg/kg bw) Reference Rat Inhal. >227 ppm (6 hr)4 Scala and Burtis, 1973 Guinea pig Inhal. >227 ppm (6 hr)4 Scala and Burtis, 1973 Rat Inhal. saturated vapors (8 hr) Smyth et al., 1969 Rat Inhal. >235 ppm Treon, 1963 (1) Gastrointestinal irritation was reported in rats after oral administration of EH undiluted or in corn oil. (2) Mild dermal irritation (3 on a scale of 0 [no irritation] to 10) was reported when EH was applied to the uncovered rabbit belly and moderate corneal injury in rabbits when undiluted EH was instilled in the eye (5 on a scale of 0 [no injury] to 10). (3) Moderate skin irritation was reported in rabbits following dermal application of EH and severe eye irritation with persistent, wide-spread corneal opacity when 0.1 ml undiluted EH was applied to the conjunctival sac. (4) Mice, rats and guinea pigs (10 each) were exposed for 6 hours to 227 ppm EH, then observed for 24 hours before necropsy. No deaths occurred during exposure or observation. All animals exposed to EH exhibited central nervous system depression and labored breathing and one guinea pig had a clonic convulsion. During exposure, mucous membranes of the eyes, nose, throat, and respiratory passages of animals exposed to EH were irritated, but animals recovered within one hour after exposure was terminated. Gross necropsy revealed areas of slight haemorrhage in animals exposed to EH. One female mouse receiving 1 000 mg 2-ethyl-1-hexanol/kg bw/day showed abdominal position and loss of consciousness; the mouse died later the same day; microscopic examination revealed tubular dilatation in the renal cortex and centrilobular fatty infiltration in the liver of this mouse. Also the following significant effects were reported to be associated with administration of 1 000 mg/kg bw/day for 11 days: 1) increased absolute stomach weights in males and females; 2) increased liver-to-bw ratio for males; 3) increased stomach-to-bw ratio for females; 4) foci in the forestomach of 3 males and 2 females; 5) hyperkeratosis and focal or multifocal acanthosis and inflammatory oedema in the submucosa of the forestomach of males and females, including focal or multifocal ulceration of the mucous membrane of a few males and females; and 6) hypertrophy of hepatocytes in one male and one female. Males (9/10) and females (6/10) receiving 1 500 mg 2-ethyl-1- hexanol/kg bw/day for 11 days had clinical signs such as ataxia and lethargy, some animals also had piloerection, and a few animals showed abdominal or lateral position and loss of consciousness; one male and four females died during the study. Microscopic evaluation showed tubular dilatation and nephrosis in the renal cortices of males and females that died intercurrently, and centrilobular fatty infiltration in the liver of females that died intercurrently. The following statistically significant effects were reported, associated with administration of 1 500 mg 2-ethyl-1-hexanol/kg bw/day: 1) increased absolute liver and stomach weights in males and females; 2) increased organ-to-bw ratios for stomach and liver in males and females; 3) increased organ-to-brain weight ratios for stomach and liver in males and females; 4) foci in the forestomach of 7/10 males and 5/10 females; 5) hyperkeratosis and focal or multifocal acanthosis and inflammatory oedema in the submucosa of the forestomach of most males and females, including focal or multifocal ulceration of the mucous membrane in a few males and females; 6) hypertrophy of hepatocytes in the liver of males and females, including focal necrosis of liver cells in one male and one female; and 7) bilateral tubular giant cells in the testicular tubules of two males (BASF, 1992b). Doses of 0, 25, 125, 250, or 500 mg 2-ethyl-1-hexanol/kg bw/day were administered by gavage to groups of 10 male and 10 female B6C3F1 mice for 3 months. Animals in the 250 and 500 mg/kg bw/day groups showed toxic effects related to administration of the test compound. For male mice receiving 250 mg/kg bw/day, statistically significant increased stomach-to-bw ratio was observed. Statistically significant effects observed in animals receiving 500 mg 2-ethyl-1-hexanol/kg bw/day included: a) increased stomach-to-bw ratio in males and 2) slight focal or multifocal acanthosis in the mucosa of the forestomach of 2/10 males and 1/10 female (BASF, 1992b). 2.2.2.2 Rats Doses of 0, 100, 330, 1 000, or 1 500 mg 2-ethyl-1-hexanol/kg bw/day were administered by gavage for 11 days (9 applications) to groups of 10 male and 10 female Fischer 344 rats. Clear toxic effects occurred in the male and female rats receiving 330, 1 000, or 1 500 mg/kg bw/day. Female rats receiving 330 mg/kg bw/day for 11 days had increased kidney-to-bw ratios, but not increased absolute kidney weights or kidney-to-brain weight ratios. Microscopic findings included inflammatory oedema in the forestomach of one female rat and decreased thymus size (microscopic examination) in 1 female and 2 male rats. Male and female rats receiving 1 000 mg 2-ethyl-1-hexanol/kg bw/day had reduced feed consumption, body weight, and body weight gain compared to control rats. Some rats in this dose group showed ataxia and apathy; a single rat showed piloerection and the genital region of one rat was smeared with urine. The following statistically significant effects were also reported to be associated with administration of 1 000 mg/kg bw/day for 11 days: a) serum cholesterol and reticulocytes in rats of both sexes were reduced; b) absolute spleen weights of rats of both sexes were reduced; c) absolute liver weights of male and female rats were increased; d) organ-to-bw ratios for stomach, liver and kidneys were increased for male and female rats; e) brain-to-bw ratio was increased in female rats; f) spleen-to-bw ratios were reduced in male and female rats; g) liver-to- brain weight ratios were increased in male and female rats; h) spleen- to-brain weight ratios were decreased in male and female rats; i) foci were reported in the forestomachs of 2 males; j) hyperkeratosis and focal or multifocal acanthosis in the mucous membrane of the forestomach of most male and female rats, as well as epithelial degeneration, ulceration and subcutaneous inflammatory oedema; k) parenchymal involution of lymphoreticular tissue in the spleens of 5 female rats; l) decreased thymus size in 2 males and 5 females (microscopic evaluation); l) lymphocyte depletion in the thymus of 5 females and lymphocyte necrosis in the thymus of 4 females. Male and female rats receiving 1 500 mg 2-ethyl-1-hexanol/kg bw/day showed reduced feed consumption, body weight, and body weight gain compared to control rats. All animals in the dose group demonstrated ataxia and lethargy, some animals showed abdominal or lateral position and appeared to be unconscious, almost all animals had piloerection, and a few rats had genital regions smeared with urine. The following statistically significant effects were also reported to be associated with administration of 1 500 mg 2-ethyl-1- hexanol/kg bw/day: a) reduced serum cholesterol, glucose, and reticulocytes in male and female rats; b) increased serum alanine aminotransferase in male rats; c) decreased absolute spleen, brain, and adrenal weights and increased absolute liver and stomach weights in male and female rats; d) increased organ-to-body weight ratios for stomach, liver, kidney, and brain in male and female rats, decreased spleen-to-bw ratios for male and female rats, increased adrenal-to-bw ratio for male rats, and increased lung-to-bw ratio for female rats; e) increased organ-to-brain weight ratios for liver and stomach in rats of both sexes, decreased spleen-to-brain weight ratio for male and female rats, and decreased adrenal-to-brain weight ratio in female rats. Foci were reported in the forestomach of 4 male and 7 female rats dosed with 1 500 mg 2-ethyl-1-hexanol/kg bw/day. Microscopic findings at this dose level were reported to include: a) hyperkeratosis and focal or multifocal acanthosis in the mucous membrane of the forestomach of all male and female rats, as well as epithelial degeneration, ulceration, and subcutaneous inflammatory oedema in some animals; b) slight hypertrophy of hepatocytes in the liver of 8 males and 8 females; c) focal hepatocellular necrosis in 1 female and 2 male rats; d) parenchymal involution of lymphoreticular tissue in the spleen of 9 male and 9 female rats; e) decreased thymus size in 10 male and 9 female rats; and f) lymphocyte depletion in the thymus of 9 male and 8 female rats and lymphocyte necrosis in the thymus of 1 male and 6 female rats (BASF, 1992a). Doses of 0, 25, 125, 250, or 500 mg 2-ethyl-1-hexanol/kg bw/day were administered by gavage to groups of 10 male and 10 female Fischer 344 rats for 3 months. Animals in the 250 and 500 mg/kg bw/day groups showed toxic effects. For animals receiving 250 mg/kg bw/day, statistically significant effects were reported to include: a) decreased serum alkaline phosphatase and glucose in male rats and decreased serum alanine aminotransferase in female rats; b) increased liver-to-bw ratios in male and female rats and increased stomach-to-bw ratio in female rats; and c) decreased fat deposition of the liver cells of male rats. Statistically significant effects observed in animals receiving 500 mg 2-ethyl-1-hexanol/kg bw/day included: a) decreased body weight and body weight gain in male and female rats; b) decreased serum alanine aminotransferase, glucose, and cholesterol in male and female rats, increased reticulocytes in male and female rats, decreased serum alkaline phosphatase in male rats, and increased serum protein and albumin in male rats; c) increased absolute liver weights in male and female rats and increased absolute stomach weights in female rats; d) increased organ-to-body weight ratios for liver and stomach in male and female rats; and e) increased organ-to-brain weight ratios for liver and stomach in male and female rats. Slightly elevated single or multiple foci were observed in the mucosa of the forestomach of male and female rats receiving 500 mg/kg bw/day. Macroscopic findings at this dose level were reported to include: a) focal or multifocal achanthosis in the mucosa of the forestomach of 1 male and 5 female rats; b) acanthosis of the whole mucosa, ballooning degeneration of the epithelia, and inflammatory oedema in the submucosa of 1 male rat; and c) decreased fat deposition in the liver and fewer animals with fatty infiltration of the lobular periphery of the liver compared to vehicle-control rats (BASF, 1992a). Five male Wistar-derived rats were administered 1 mmol/kg/day (approximately 130 mg/kg bw/day) EH dissolved in polyethylene glycol 300 (10 ml/kg/day) by gavage for 14 days; 10 control rats were administered the gavage vehicle alone. At the end of the treatment period, rats were killed and blood was withdrawn for analysis of plasma cholesterol and triglyceride levels; livers and testis were weighed, liver samples were taken for light and electron microscopy, and the remaining liver was homogenized for determination of total catalase and CN-insensitive palmitoyl CoA oxidation. No major pathological signs of hepatotoxicity were observed, although slight centrilobular hypertrophy (controls: 4/10 rats; EH-treated: 2/5), slight/moderate glycogen vacuolization (controls: 9/10; EH-treated: 5/5), and slight/moderate centrilobular "fat" vacuolation (controls: 9/10; EH-treated: 1/5) were reported in control and EH-treated rats. Administration of EH had no significant effect on body weight gain, liver-to-body-weight ratio, testis-to-body-weight ratio, number of peroxisomes/504 µm2 liver, serum catalase activity, serum cholesterol, or serum triglycerides. In addition, 0.1 mM EH had no effect on acyl CoA oxidase activity after 72 hrs in in vitro culture with rat hepatocytes (Rhodes et al., 1984). Gavage administration of 1335 mg/kg bw/day EH in corn oil to 6 male Wistar albino rats for 7 days resulted in significantly increased liver-to-body-weight ratio (control: 3.5±0.1 g/100 g bw; EH-treated: 4.9±0.1 [p<0.001]), decreased glucose-6-phosphatase activity (control: 24±2 µg/min/mg microsomal protein; EH-treated: 15±1 [p<0.01]), increased biphenyl 4-hydroxylase activity (control: 1.6±0.1 µmol/hr/g liver; EH-treated: 2.1±0.1 [p<0.01]), and increased microsomal cytochrome P-450 content (control: 0.07±0.003 delta-E 450-500 nm/mg microsomal protein; EH-treated: 0.1±0.004 [p<0.001]) (Lake et al. 1975). 2.2.3 Long-term toxicity/carcinogenicity studies 2.2.3.1 Mice 2-ethyl-1-hexanol (0, 50, 200, or 750 mg/kg bw/day) was administered by gavage to groups of 50 male and 50 female B6C3F1 mice five days per week for a period of 18 months. The purity of the test substance was reported to be greater than 99.87%. The gavage vehicle was doubly distilled water containing 5 mg Polyoxyl 35 castor oil per 100 ml. An additional control group of 50 male and 50 female rats was gavaged with double distilled water only. All rats received 10 ml/kg bw test substance emulsion, vehicle or double distilled water per dose. The method of preparing test substance emulsions was changed after 6 months of dosing because homogeneity analyses of emulsions showed considerable variation, ranging from approximately 70% to 140% of target concentrations. However, variability in dosing during the early part of the study did not significantly affect the outcome of the study because of the "clear biological distinction between dose levels during treatment." Mice were housed singly and feed and water were available ad libitum throughout the study. At the initiation of dosing, mice were 49 days old; mean body weight of males was 23 g (range 21-26 g) and mean body weight of females was 19 g (range 17-23 g). At the end of the study, non-fasted mice were decapitated under CO2 anaesthesia. The general health of the test animals was checked daily, and test animals were examined and palpated once a week. Mice were weighed weekly during the first 13 weeks, then every four weeks for the duration of the study. Feed consumption was determined for a period of one week every four weeks during the study. Blood samples were drawn from the tail vein of all surviving animals for haematological examination at 12 months and at the end of the study. Animals that survived to the end of the study were necropsied; tissues and organs were subjected to gross and microscopic pathology examination. No EH-related changes were observed in mice administered 50 or 200 mg/kg bw/day EH for 18 months. In mice administered 750 mg/kg bw/day, the following effects were observed: 1) decreased body weight gain in males (approximately 26%) and females (24%) that was associated with a substantial reduction in feed consumption (males: decreased from about 9% to 20%; females: decreased from about 9% to 30%); 2) increased mortality in males (vehicle controls: 4%; EH-treated: 30%) and females (vehicle controls: 8%; EH-treated: 30%); 3) treatment-related haematological changes, including slightly increased polymorphonuclear neutrophils in males and females (males: controls--19.8±5.9% [12 mo] and 20.5±7.6% [18 mo]; 750 mg/kg bw/day--26.4±9.1% [12 mo] and 26.9±13.5% [18 mo])(females: controls-- 20.6±5.6% [12 mo] and 22.6±10.7% [18 mo]; 750 mg/kg bw/day--24.2±5.4 [12 mo] and 25.1±9% [18 mo]) and slightly decreased lymphocytes in males and females (males: controls--77±6.9% [12 mo] and 76.6±7.6% [18 mo]; 750 mg/kg bw/day--69.1±9% [12 mo] and 70.6±13.2% [18 mo]); and 4) treatment-related, but not statistically significant, increased focal hyperplasia of the epithelium of the forestomach in males (controls-- 1/50; 50 mg/kg bw/day--1/50; 200 mg/kg bw/day--1/50; 750 mg/kg bw/day--5/50) and females (controls--1/50; 50 mg/kg bw/day--1/50; 200 mg/kg bw/day--0/50; 750 mg/kg bw/day--4/50). Also, a slight increase in the incidence of hepatocellular carcinomas in high-dose females was statistically significant when compared to the incidence in vehicle control females but not when compared to the incidence in water- gavaged control females (vehicle control--0/50; 50 mg/kg bw/day--1/50; 200 mg/kg bw/day--3/50; 750 mg/kg bw/day--5/50). No statistically significant increase in tumour incidence occurred in male mice. EH is not oncogenic in the mouse under the conditions of this study (BASF, 1992b). In a satellite study to the carcinogenicity study in mice cited above as BASF, 1992b, EH was administered by gavage (vehicle: distilled water containing 5 mg Polyoxyl 35 castor oil per 100 ml) to two groups of male and female B6C3F1 mice at 750 mg/kg bw/day. A control group of 10 males and 10 females was gavaged with the vehicle only for 13 months; a second (non-recovery) group of 10 males and 10 females was gavaged with EH for 13 months, 5 days/week; a third (recovery) group of 50 males and 50 females was gavaged with EH for 13 months, 5 days/week, then gavaged with the vehicle only for 5 months, 5 days/week. Mice were killed at the end of the treatment periods--13 months for groups one and two; 18 months for group three--and subjected to gross pathological assessment. The general health of the test animals was checked daily, and animals were examined and palpated once a week. Body weights were determined once a week during the first 13 weeks of the study, then once every four weeks. Feed consumption was determined one week in every four weeks throughout the study. Microscopic examination of tissues and organs was performed only on mice that died during the study. Administration of 750 mg/kg bw/day EH to male and female mice for 13 months caused increased mortality in males and females (males: control--0%, non-recovery--30%, recovery--22% during the first 13 months; females: control--0%, non-recovery--20%, recovery--16% during the first 13 months). For the non-recovery mice and recovery mice during treatment with EH, feed consumption was significantly decreased compared to control mice (at 13 months, males: control--4.7±0.4 g/day, non-recovery--3.7±0.5 g/day, recovery--4.4±0.6 g/day; females: control--6.0±1.3 g/day, non-recovery--5.6±1.5 g/day, recovery--5.8±1.0 g/day); following the 5 month recovery period, feed consumption for mice in the recovery group was in the same range as feed consumption for control mice (males: control--4.8±0.9 g/day, recovery--4.9±0.8 g/day; females: control--6.3±1.3 g/day, recovery--5.9±1.1 g/day). For the non-recovery mice and recovery mice during treatment with EH, body weight gain was significantly decreased (at 13 months, males: control--40.9±2.7 g, non-recovery--36.7±2.8 g, recovery--38.7±3.2 g; females: control--38.7±5.4 g, non-recovery--33.8±4.4 g, recovery-- 34.9±4.8 g); following the 5 month recovery period, body weight gain of female mice that had been gavaged with EH for 13 weeks was still significantly decreased compared to control mice (males: control-- 42.7±3.5 g, recovery--42.4±3.6 g; females: control--41.1±5.6 g, recovery--36.9±5.1 g). Some statistically significant changes in organ weights and masses or foci in liver and stomach were observed to be associated with EH administration; these were similar to changes noted in the results of the carcinogenicity study (BASF, 1992d). 2.2.3.2 Rats 2-ethyl-1-hexanol (0, 50, 100, or 150 mg/kg bw/day) was administered by gavage to groups of 50 male and 50 female Fischer 344 rats five days/week for a period of 24 months. The purity of the test substance was reported to be greater than 99.3%. The gavage vehicle was doubly distilled water containing 5 mg Polyoxyl 35 castor oil per 100 ml. An additional control group of 50 male and 50 female rats was gavaged with double distilled water only. All rats received 10 ml/kg bw/day test substance emulsion, vehicle or double distilled water per dose. The method of preparing test substance emulsions was changed after 6 months of dosing because homogeneity analyses of emulsions showed considerable variation, ranging from approximately 70% to 140% of target concentrations. However, the report concluded that variability in dosing during the early part of the study did not significantly affect the outcome of the study because of the "clear biological distinction between dose levels during treatment." Rats were housed singly and food and water were available ad libitum throughout the study. At the initiation of dosing, rats were 42 days old; mean body weight of males was 103 g (range 86-128 g) and mean body weight of females was 81 g (range 64-95 g). At the end of the study, non-fasted rats were decapitated under CO2 anaesthesia. The general health of the test animals was checked daily, and test animals were examined and palpated once a week. Rats were weighed weekly during the first 13 weeks, then every four weeks for the duration of the study. Feed consumption was determined for a period of one week every four weeks during the study. Blood samples were drawn from a tail vein of all surviving animals for haematological examination at 12 and 18 months and at the end of the study. At the end of the study, surviving animals were necropsied; tissues and organs were subjected to gross and microscopic pathology examination. No compound-related changes were associated with administration of 50 mg/kg bw/day for 24 months; however, body weights and body- weight gains of rats receiving 50, 150, or 500 mg 2-ethyl-1-hexanol/kg bw/day were decreased in a statistically significant dose-dependent manner compared to vehicle control rats. At the end of the study, body weights were about 5%, 11%, and 23% below control values and body weight gains were about 8%, 16%, and 33% below control values, respectively. Feed consumption of male and female rats receiving 500 mg/kg bw/day showed occasional statistically significant decreases compared to both control groups of rats, but no dose-response relationship was observed. An EH-associated increase in mortality was observed for female mice of the high-dose group only (males: vehicle controls--34%, 50 mg/kg bw/day--46%, 150 mg/kg bw/day--32%, 500 mg/kg bw/day--38%; females: vehicle controls--28%, 50 mg/kg bw/day--28%, 150 mg/kg bw/day--26%, 500 mg/kg bw/day--52%). For rats receiving 100 mg/kg bw/day, the study reported a) statistically significant reductions in body weight (males: 11%; females: 9%) and body weight gain (males: 16%; females: 12%) compared to vehicle control rats and b) slightly increased numbers of animals with clinical symptoms and incidences of symptoms (frequency/animals) such as poor general condition (100 mg/kg bw/day: males--69/15; vehicle control: males--62/12), labored breathing (100 mg/kg bw/day: males--4/1, females--30/5; vehicle control: males--2/1, females--9/3), piloerection (100 mg/kg bw/day: males--17/1; vehicle control: males-- 0/0), and genital regions smeared with urine (100 mg/kg bw/day: females--31/4; vehicle controls: females--0/0). Feed consumption of male and female rats receiving 150 mg/kg bw/day showed occasional statistically significant decreases compared to both control groups of rats, but no dose-response relationship was observed. The following treatment-related changes were observed in rats dosed with 500 mg/kg bw/day 2-ethyl-1-hexanol for 24 months: a) statistically significant reductions in body weight gain for males (33%) and females (31%); b) increased incidences of male and female rats with clinical symptoms (frequency/animals) such as poor general condition (500 mg/kg bw/day: males--200/14, females--248/21; vehicle control: males--62/12, females--34/8), labored breathing (500 mg/kg bw/day: males--41/4, females--75/12; vehicle control: males--2/1, females--9/3), piloerection (500 mg/kg bw/day: males--67/2, females-- 21/5; vehicle control: males--0/0, females--2/1), and/or genital region smeared with urine (500 mg/kg bw/day: males--13/1, females-- 502/21; vehicle control: males--0/0, females--44/6); and c) statistically significant increased mortality in dosed females as reflected in the number of animals that died or were sacrificed in a moribund condition during the study (52%) compared with vehicle control females (28%). Male rats dosed with 500 mg/kg bw/day had slightly increased anisocytosis, predominantly microcytosis at 12 months, but not at 18 nor 24 months, compared to vehicle control males. No malignant tumours were detected in high-dose animals that died before scheduled termination and the sum of primary tumours, benign tumours and malignant tumours was remarkably lower in the high-dose group compared to both control groups of rats. Thus, 2-ethyl-1-hexanol was not oncogenic in the rat under conditions of this assay (BASF, 1992a). In a satellite study to the BASF carcinogenicity study in rats cited above as BASF 1992a, EH was administered by gavage (vehicle: distilled water containing 5 mg Polyoxyl 35 castor oil per 100 ml) to two groups of male and female Fischer 344 rats at 500 mg/kg bw/day. A control group of 10 males and 10 females was gavaged with the vehicle only for 18 months; a second (non-recovery) group of 10 males and 10 females was gavaged with EH for 18 months, 5 days/week; a third (recovery) group of 50 males and 50 females was gavaged with EH for 18 months, 5 days/week, then gavaged with the vehicle only for 6 months, 5 days/week. Rats were killed at the end of the treatment periods--18 months for groups one and two; 24 months for group three--and subjected to gross pathological assessment. The general health of the test animals was checked daily, and animals were examined and palpated once a week. Body weights were determined once a week during the first 13 weeks of the study, then once every four weeks. Feed consumption was determined one week in every four weeks throughout the study. Microscopic examination of tissues and organs was performed only on rats that died during the study. Administration of 500 mg/kg bw/day EH to male and female rats for 18 months caused slightly increased mortality in females (control-- 20%, non-recovery--40%, recovery--34% during the first 18 months) and decreased feed consumption in males (maximum decrease of approximately 12%). For the non-recovery rats and recovery rats during treatment with EH, body weight gain was significantly decreased (at 18 months, males: control--298.7±19.6 g, non-recovery--215.4±21.3 g, recovery--211.2±22.4 g; females: control--149.9±19.8 g, non- recovery--128.7±21.7 g, recovery--128.5±18.3 g); following the 6 month recovery period, body weight gains of males and females that had been gavaged with EH for 18 weeks had partially recovered but were still significantly decreased compared to controls (males: control-- 266.5±30.7 g, recovery--216.1±23.0 g; females: control--177.5±27.0 g, recovery--150.6±21.5 g). The following changes were observed in rats that had been gavaged with 500 mg/kg bw/day EH for 18 months compared to control rats: 1) a greater number of animals and/or a higher incidence of clinical symptoms such as poor general condition, labored breathing, and genital region smeared with urine in males and females; 2) statistically significant decreases in the absolute weights of brain (males and females) and stomach (males); 3) statistically significant decreases in organ-to-body weight ratios of brain (males and females), liver (males and females), kidneys (males and females), stomach (males and females), and testes (BASF, 1992c). 2.2.4 Reproduction studies The response of mixed cultures of Sertoli and germ cells prepared from Sprague-Dawley rat testes to model testicular toxicants was studied. After incubation of the cultures with 2 x 10-4 M EH for 24 hrs, no increase was observed in the normal rate of germ cell detachment from Sertoli cells into the culture medium (Gray and Beamand 1984). Effects of EH on rat testes were examined in vivo and in vitro. No testicular damage was observed in male Sprague-Dawley rats given oral doses of 2.7 mmol EH/kg bw/day for 5 days and incubation with EH (0-1 000 µM for 24 or 48 hours) did not enhance detachment of germ cells from primary mixed cultures of rat Sertoli and germ cells (Sjoberg et al., 1986). EH did not increase lactate and pyruvate concentrations in the medium of in vitro cultures of rat Sertoli cells. Such increases are considered to be sensitive indicators of altered Sertoli cells function associated with Sertoli-cell toxicants (Williams & Foster 1988). 2.2.5 Special studies on developmental toxicity and teratogenicity 2.2.5.1 Mice Pregnant CD-1 mice were gavaged on gestation days 6-13 with 1525 mg/kg bw/day EH in corn oil; control mice were gavaged with corn oil; dams were allowed to litter. Administration of EH caused statistically significant (p<0.05) decreased maternal body weight gain (control: 7.0±2.5 g; EH: 3.9±3.2 g), decreased number of viable litters (control: 33/34; EH: 11/20), decreased liveborn per litter (control: 9.9±2.4; EH: 6.8±3.4), decreased percentage survival of pups (control: 98.2±8.8; EH: 73.4±32.2), and decreased birth weight (control: 1.6±0.1 g/pup; EH: 1.4±0.2 g/pup) and weight gain for pups (control: 0.6±0.1 g/pup; EH: 0.3±0.2 g/pup) (Hardin et al., 1987). 2.2.5.2 Rats Pregnant Wistar rats were administered undiluted di(2-ethylhexyl) phthalate (DEHP; 12.5 or 25 mmol/kg bw), EH (6.25 or 12.5 mmol/kg bw, approximately equivalent to 800 and 1600 mg/kg bw), or 2-ethylhexanoic acid (EA; 6.25 or 12.5 mmol/kg bw) by gavage on day 12 of gestation. Control rats were not gavaged (untreated controls). Caffeine (150 mg/kg) was dissolved in water and injected i.p. in some pregnant rats of each group. Rats were killed on day 20 of gestation; following Caesarean section, implantation sites were determined in situ and the number of dead or resorbed fetuses was determined. Live fetuses were removed and examined; internal and external soft tissue and skeletal malformations were recorded. At least seven litters for each experimental condition were analyzed. Administration of each test compound resulted in statistically significant, dose-related increases in malformed live fetuses (DEHP: 12.5 mmol/kg bw [7 litters]--4.5±4.5%, 25 mmol/kg bw [7 litters]-- 20.8±7.3%; EH: 6.25 mmol/kg bw [7 litters]--2.0±1.3%, 12.5 mmol/kg bw [7 litters]--22.2±14.7%; EA: 6.25 mmol/kg bw [7 litters]--0.8±0.8%; 12.5 mmol/kg bw [10 litters]--67.8±10.9%) compared to controls (no malformed live fetuses in 7 litters). Defects in fetuses following treatment with EH included hydronephrosis (7.8% of live fetuses), tail defects (4.9% of live fetuses), limb defects (9.7% of live fetuses), and other defects (1.0% of live fetuses). For each test compound, caffeine was reported to potentiate (increase) the percent of malformed live fetuses. However, administration of test compounds did not significantly affect the percentage of dead and resorbed fetuses compared to controls. No maternal effects associated with the test compounds were reported. These results are consistent with the hypothesis that the proximal teratogen for DEHP is EA, the metabolic product of EH (Ritter et al., 1986 and 1987). The developmental toxicity of dermally applied EH was studied in Fischer 344 rats; results of a dose range-finding study for the developmental toxicity study were also included. In the dose range- finding study 0, 420, 840, 1680, or 2520 mg/kg bw/day EH (undiluted) was applied to the clipped dorsal skin of pregnant F344 rats (8 rats/group); a positive dermal control group (2-methoxyethanol) and a sham-treated (deionized water) dermal control group were included in the study. In the developmental toxicity study, 0, 252, 840, or 2520 mg/kg bw/day EH (undiluted) was applied to the clipped dorsal skin of pregnant F344 rats (25/group); a positive dermal control group (2-methoxyethanol) and a sham-treated (deionized water) dermal control group also were included in this study. Body weights were recorded on gestation days 0, 6, 9, 12, 15, and 21; feed consumption was estimated for 3-day intervals from gestation days 0-21. Skin irritation was measured before and after each 6-hr application period. Surviving females were killed on gestation day 21; uterine and liver weights (both studies) and weights of spleen, adrenals, kidneys, and thymus (developmental toxicity study) were recorded. Corpora lutea and uterine implantation sites were counted; ovaries, cervices, vaginas, and abdominal and thoracic cavities were examined grossly. All live and dead fetuses and resorption sites were noted. Live fetuses were sexed, weighed, and examined for external, visceral, and skeletal malformations and variations. All pregnant females treated with EH survived. Clinical findings for EH-treated pregnant rats were limited to body weight changes, skin irritation, and nasal and ocular effects. Decreased body weight gain was observed in the dose range-finding study for gestation days 6-15 at doses of 1 680 (10.1±7.1 g) and 2 520 mg/kg bw/day EH (10.7±4.8 g) compared to sham-treated control rats (18.9±6.4 g). In the main study, weight gain was statistically significantly decreased for gestation days 6-9 at 2 520 mg/kg bw/day EH (0.1±2.4 g) compared to sham-treated controls (3.3±1.2 g), and was somewhat, but not statistically significantly, decreased at 840 mg/kg bw/day EH. No significant changes in feed consumption were reported at any treatment level of EH in either study throughout gestation (data not given). EH-related irritation effects at the treatment site were identified as mild, and included exfoliation, encrustation and erythema for all treatment groups in both studies; oedema was not observed. Gestational effects were observed for neither study at any dose of EH applied dermally. Also, dermal administration of EH was not associated with external, visceral, or skeletal malformations. Dermally applied EH does not produce developmental or teratogenic effects when administered at doses associated with demonstrable maternal toxicity (Tyl et al., 1992). Groups of approximately 15 pregnant Sprague-Dawley rats were exposed for 7 hrs/day to air saturated with EH vapor (approximately 850 mg/m3 EH) throughout gestation (Nelson et al., 1988). Dams were weighed daily during the first week of exposure, then weekly. Dams were killed on gestation day 20; fetuses were removed, sexed, weighted and examined for external, visceral and skeletal defects. EH reduced maternal feed intake but did not produce significant maternal toxicity (data not provided). Inhalation of EH under conditions of this experiment was not associated with increased malformations (Nelson et al., 1988). 2.2.6 Special studies on genotoxicity The results of genotoxicity assays on EH are summarized in Table 2. All reports except Seed (1982) were of negative results for in vitro assays; results were negative for several in vivo assays, including a dominant lethal assay, a chromosomal aberration assay, and a mutagenicity assay on rat urinary metabolites of EH. 2.2.7 Observations in humans Hollenbach and coworkers (1972) reported that laboratory workers exposed to EH (among other substances) reported headaches, dizziness, fatigue and gastrointestinal disorders; also that exposed workers had slightly decreased blood pressure during the day. Table 2: Results of genotoxicity assays on EH Test Test Subject EH Conc. Result Reference Ames test1 S. typhimurium TA98 0-1.0 µL/plate neg. Kirby et al., 1983 TA100 TA1535 TA1537 TA1538 Ames test1 S. typhimurium TA98 0-220 µg/plate neg. Zeiger et al., 1985 TA100 TA1535 TA1537 Ames test1 S. typhimurium TA98 0-2 000 µg/plate neg.3 Agarwal et al., 1985 TA100 TA1535 TA1537 TA1538 TA2637 Ames test1 S. typhimurium TA98 0-1.8 µl/plate neg. Litton Bionetics Inc., 1982a TA100 TA1537 TA1535 TA1538 Ames test1 S. typhimurium TA98 urine from rats neg. DiVincenzo et al., 1983 TA100 TA1535 TA1537 gavaged with 1 g/kg TA1538 bw/day EH for 15 day In vitro cell BALB/3T3 cells 0-0.162 µg/ml neg.4 Litton Bionetics Inc., 1982b transformation assay1 8-Azaguanine S. typhimurium TA100 0-1.5 mM pos.5 Seed, 1982 resistance assay2 Mouse micronucleus B6C3F1 mouse bone 456 mg/kg bw/day neg.6 Litton Bionetics Inc., 1982c test marrow cells i.p. for 1 or 2 day Table 2 (contd) Test Test Subject EH Conc. Result Reference mouse lymphoma L5178Y/TK+/- mouse 0.01-0.24 µL/mL neg. Kirby et al., 1983 assay1 lymphoma cells Rec-assay Bacillus subtilis 500 µg/disk neg. Tomita et al., 1982 CHO mutation assay Chinese hamster ovary 1.5-2.8 mM neg. Phillips et al., 1982 (CHO) cells Unscheduled DNA Primary rat hepatocytes Not given neg. Hodgson et al., 1982 synthesis assay In vivo dominant ICR/SIM mice 250, 500, 1 000 neg. Rushbrook et al., 1982 lethal assay mg/kg bw/day for 5 day In vivo chromosomal F344 rat bone marrow cells .02, .07, .21 g/kg neg. Putnam et al., 1983 aberration assay bw/day for 5 day (1) Both with and without metabolic activation (2) Without metabolic activation (3) Moderate cytotoxicity reported in most cultures (4) Negative from 0-225 µl/ml without metabolic activation; negative from 0-0.162 µl/ml with rat hepatocytes for metabolic activation (5) Small dose-related increase (maximum increase was approximately 3.5 times background) in mutation frequency accompanied by decreased survival (cytotoxicity) (6) Negative with and without activation with S9 and with and without ß-glucuronidase/arylsulfatase 3. COMMENTS In rats, orally administered 2-ethyl-1-hexanol is absorbed and rapidly eliminated within 28 hours, mainly in urine and faeces. The major urinary metabolite is 2-ethylhexanoic acid. In mice, rats and monkeys, the compound is oxidized by ß-, omega-, and omega-1-oxidation to various metabolites, including 2-ethylhexanoic acid, ethylhexanedioic acid, and 5-hydroxyethylhexanoic acid. Glucuronic acid conjugates are formed in all three species. The Committee concluded that the available data do not indicate that 2-ethyl-1-hexanol is genotoxic. With a single exception, in which a positive result occurred in the presence of significantly decreased cell survival (cytotoxicity), the results of both in vivo and in vitro genotoxicity tests were negative. Although teratogenic effects were reported in the offspring of mice administered 1 500 mg 2-ethyl-1-hexanol/kg bw/day by gavage on days 6-13 of gestation, these effects occurred in the presence of severe maternal toxicity. The body weight gain of treated females was approximately 40% less than that of untreated controls. In rats, administration of 1600 mg/kg bw 2-ethyl-1-hexanol by gavage (but not 800 mg/kg bw) on day 12 of gestation was associated with a statistically significant increase in the number of malformed live fetuses (malformations included hydronephrosis, tail defects and limb defects). Maternal toxicity was not reported in this study. The results of several short-term toxicity studies suggested that 2-ethyl-1-hexanol administered orally to rats and mice at doses greater than approximately 350 mg/kg bw/day induces liver peroxisome proliferation and/or marker enzymes for peroxisome proliferation. However, the results of carcinogenicity studies did not indicate that long-term oral administration of 2-ethyl-1-hexanol leads to induction of liver tumours in mice or rats. The results of long-term oral carcinogenicity studies indicated that 2-ethyl-1-hexanol is not carcinogenic in rats (24 months) or mice (18 months). The incidence of hepatocellular carcinomas at 750 mg/kg bw/day in female mice was slightly higher than in historical controls, however this effect was considered to be incidental and unrelated to the administration of 2-ethyl-1-hexanol. The increase was statistically significant when compared with the incidence in vehicle control females but not when compared with the incidence in control females given distilled water by gavage. In these studies, the 750 mg/kg bw/day dose of the compound produced a number of statistically significant, non-carcinogenic adverse effects, but these effects were not observed at 50 or 200 mg/kg bw/day in mice or at 50 mg/kg bw/day in rats. 4. EVALUATION On the basis of a NOEL of 50 mg/kg bw/day from the long-term study in rats and using a safety factor of 100, the Committee established an ADI of 0-0.5 mg/kg bw for 2-ethyl-1-hexanol. 5. REFERENCES AGARWAL, D.K., LAWRENCE, W.H., NUNEZ, L.J. & AUTIAN, J. (1985). 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See Also: Toxicological Abbreviations