International industry initiatives to improve the glycol ether health effects knowledge base.

Following the recognition in the early 1980s of the potential reproductive hazards of certain glycol ethers, industry organizations were formed in the US and Europe having a number of stated goals to: (1) provide hazard information by expanding the toxicity database for glycol ethers; (2) promote cooperation among scientists, governmental authorities, and industry; and (3) promote scientifically sound regulatory actions and to assist in the setting of scientifically defensible safety standards. This effort led to early recommendations that EGME, EGEE, and their acetates be removed from consumer products. Also, studies conducted by industry under US EPA test rules have led to a better understanding of the hazards associated with glycol ether constituents of brake fluids, paints, and other products. Industry-provided information has greatly assisted the setting of occupational and public safety standards in a number of countries. Hazard assessments for a number of large-volume glycol ethers have been performed under the OECD SIDS program. This work continues with the industry-funded ICCA/HPV testing initiative. To provide sound risk assessment data, industry continues to sponsor basic research aimed at better understanding human versus mouse versus rat sensitivities to certain glycol ethers. Industry has also prepared and supported the publication of toxicological data compendia for glycol ethers.

In vivo metabolism and kinetics of ethylene glycol monobutyl ether and its metabolites, 2-butoxyacetaldehyde and 2-butoxyacetic acid, as measured in blood, liver and forestomach of mice.

1. Ethylene glycol monobutyl ether (EGBE) causes forestomach hyperplasia and neoplasia in mice when administered chronically by inhalation. 2. The study was initiated to test the physiologically based pharmacokinetic (PBPK) model prediction that 2-butoxyacetaldehyde (BAL), a transient, labile intermediate in the oxidation of EGBE to butoxyacetic acid (BAA), is unlikely to achieve concentrations sufficient to cause DNA damage in target tissues. 3. Male and female B6C3F1 mice were administered a high oral dose of EGBE (600mgkg(-1)), and tissues were collected at 5, 15, 45 and 90min following the dose. The tissues were processed for determination of EGBE, BAL and BAA by gas chromatography-mass spectrometry. 4. BAL was detected at low concentrations in all tissues sampled and at all time points following EGBE administration (about 0.3-33 microM). BAL concentrations were highest in the initial samples (5 min) in all tissues and declined from that point. 5. BAL concentrations in liver and forestomach tissues corresponded to the peak concentrations predicted by an already published PBPK model, and are higher than BAL concentrations that could be achieved by inhalation exposure to EGBE. 6. Mouse inhalation exposure to EGBE is therefore unlikely to generate BAL concentrations in tissues sufficient to initiate a carcinogenic response.

Covalent protein adducts of hydroquinone in tissues from rats: identification and quantitation of sulfhydryl-bound forms.

The Michael-type addition of sulfhydryl groups to benzoquinone (BQ) or substituted benzoquinones is proposed as the primary mechanism by which these electrophilic intermediates react with either cellular glutathione or protein sulfhydryls. This reaction constitutes a reductive alkylation with a substituted hydroquinone (HQ) derivative resulting from the addition. In the case of HQ, oxidative conversion of the parent material to BQ followed by conjugation with glutathione leads to metabolic activation, producing intermediates which are nephrotoxic as well as having other proposed biological activities. Chemically, BQ may react with more than 1 equiv of glutathione (or other sulfhydryl reagents) to produce HQ derivatives substituted with up to four sulfhydryl groups. Similarly, multiply substituted protein-S adducts of HQ were anticipated to occur in vivo following administration of this material. In the current studies, sulfhydryl-bound HQ protein adducts were detected and quantitated in protein isolated from rats using a modification of the alkaline permethylation procedure of Slaughter and Hanzlik [(1993) Anal. Biochem. 208, 288-295]. In particular, total protein-S adducts to HQ in kidney or blood reached a level of 420 or 80 pmol/mg of protein, respectively, 6 h following a single gavage dose of 100 mg/kg HQ. Measured half-lives of protein-S adducts in kidney and blood were 23.9 and 36.0 h, respectively. The applicability of protein-S adducts as a tissue dosimeter for HQ is discussed.

Covalent protein adducts of hydroquinone in tissues from rats: quantitation of sulfhydryl-bound forms following single gavage or intraperitoneal administration or repetitive gavage administration.

The current studies were conducted to investigate the degree and type of protein binding of hydroquinone (HQ) in the rat following single oral or intraperitoneal (ip) or repeated oral administrations. Male or female F-344 rats or male SD rats received a single dose of HQ at 0, 25, 50, or 100 mg/kg by either gavage or ip injection (SD rats only). In addition, male or female F-344 or male SD rats received HQ by gavage for 6 weeks (5 days/week) at 0, 25, or 50 mg/kg/day. Sulfhydryl-bound HQ was quantitated in protein from blood, kidneys, livers, or spleens 24 h after treatment using an alkaline permethylation procedure. The amount of total protein-S adducts increased with increasing dose in all the tissues that were assayed. Female rats had higher levels of adducts in blood, livers, and kidneys than did male rats when they were treated orally. Male F-344 rats treated orally had elevated levels of adducts in these same tissues compared to SD rats treated orally. For all genders and strains of rats and for all treatment regimens, mono-adducts predominated in livers (>72% of total). In the kidneys, tri- and tetrasubstituted adducts predominated with the summation accounting for >60% of the total. Ip administration of HQ resulted in significantly elevated levels of adducts in all the tissues that were examined, with the greatest increases seen for protein from blood and spleens. Levels of protein-S adducts of HQ in rat kidney following a single gavage administration correlated well with previously published differences in acute HQ nephrotoxicity in rats (female F-344 rat > male F-344 rat > male SD rat). Elevated levels of HQ protein-S adducts following repeated gavage administration did not correlate to measurable clinical signs of nephrotoxicity. Evidence is presented suggesting a possible role for the prostaglandin H synthase complex in the metabolic activation of HQ. In addition, protein arylation alone cannot account for the greater sensitivity of male F-344 rats toward chronic administration of HQ. The sensitivity of male F-344 rats to HQ is likely due to other factors, including the incidence and severity of chronic progressive nephropathy.

Acute toxicity of ethylene glycol mono-n-butyl ether in the guinea pig.

Acute toxicity values, such as oral and percutaneous LD50s, are often used as the basis for classifying chemicals into toxicity categories, and their subsequent regulation. Such values obtained for ethylene glycol mono-n-butyl ether (EGBE; 2-butoxyethanol) in rats and rabbits indicate that it is moderately toxic. However, the cause of death in these acute studies appeared to be secondary to acute intravascular haemolysis, an effect for which guinea pigs and humans are much less sensitive than rats, mice and rabbits. Recently-conducted acute toxicity studies in the guinea pig resulted in an acute oral LD50 of 1400 mg/kg, an acute percutaneous LD50 of greater than 2000 mg/kg, and a 1-hr LC50 greater than 633 ppm. These data are compared with published acute toxicity values, and indicate that the predicted acute toxicity of EGBE in humans, based on data from the guinea pig, would be less than that observed in other animal species. Based in part on the guinea pig data, EBGE is no longer classified as a poisonous substance by either the United Nations or US Department of Transportation.

Dermal absorption and pharmacokinetics of isopropanol in the male and female F-344 rat.

Isopropanol (IPA), as a 70% aqueous solution, was applied under occluded conditions to the shaved backs of male and female Fischer F-344 rats for a period of 4 hr. Maximum analyzed blood concentrations of IPA were attained at 4 hr and decreased steadily following removal of the test material. Blood concentrations were below the limit of quantification at 8 hr. Acetone (ACE) blood levels rose steadily during the 4-hr exposures and continued to rise following removal of the test material, reaching peak analyzed levels at 4.5 hr (male) and 5 hr (females). ACE blood concentrations were below the limit of quantification at 24 hr. Basic pharmacokinetic parameters were similar for male and female rats with mean, first-order elimination half-lives for IPA and ACE of 0.8 to 0.9 hr and 2.1 to 2.2 hr, respectively. Following iv administration of [14C]IPA, 50-55% of the dose was eliminated as 14CO2 with lesser amounts recovered as expired volatiles or in urine. Total recoveries following iv administration were 83% for both males and females. Following a 4-hr dermal exposure to [14C]IPA (70% aqueous solution), 84-86% of the dose was recovered from the application site. Dermal absorption rates were calculated by two independent methods. The values obtained were 0.78 +/- 0.03 and 0.85 +/- 0.04 mg/cm2/hr for males and 0.77 +/- 0.13 and 0.78 +/- 0.16 mg/cm2/hr for females. Calculated permeability coefficients of 1.37 to 1.50 x 10(-3) cm/hr for males and 1.35 to 1.37 x 10(-3) cm/hr for females indicate that in the rat, IPA is rapidly absorbed dermally when applied under occluded conditions.

Ethylene glycol ethers: an environmental risk assessment.

Ethylene glycol ethers and acetates are used as intermediates, solvents, and plasticizers. They primarily enter the environment from manufacturing effluents and emissions and during their use in commercial products. Therefore, an examination of their ultimate fate and toxicity, as well as their potential for exposure, was performed. Overall, these data show that ethylene glycol ethers and acetates are not persistent in the environment, are not bioaccumulative, are generally classified by U.S. Environmental Protection Agency (EPA) procedures as "practically non-toxic" to aquatic organisms based on acute toxicity, and that conservatively calculated exposures are mostly below concentrations of concern for chronic risks to aquatic life.

Differences in the nephrotoxicity of hydroquinone among Fischer 344 and Sprague-Dawley rats and B6C3F1 mice.

The present studies indicate pronounced species-, sex-, and strain- related differences in the acute nephrotoxicity of hydroquinone (HQ) when administered by gavage to male and female Sprague-Dawley (SD) rats, Fischer 344 (F344) rats, and B6C3F1 mice. Following a single dose of 400 mg/kg, male and female F344 rats displayed pronounced enzymuria and glucosuria. In female F344 rats, urinary alkaline phosphatase and glucose were the most sensitive indicators of renal toxicity, reaching levels of, respectively, 157 times and 137 times control values within 24 h of dosing. HQ treatment of male F344 rats also resulted in significant enzymuria, although it was less marked than that seen in female F344 rats. Significant numbers of epithelial cells were also present in the urine from F344 rats at 200 (female) or 400 mg/kg (male and female). SD rats did not show evidence of elevated levels of urinary enzymes or increased blood urea nitrogen (BUN) after oral administration of HQ at a dose level of 400 mg/kg. Oral administration of HQ to male and female B6C3F1 mice at 350 mg/kg resulted in only slight but significant increases in BUN.

Metabolism of 2-ethylhexanol administered orally and dermally to the female Fischer 344 rat.

1. Excretion balance studies were conducted with 2-ethylhexanol (2-EH) in female Fischer 344 rats following single high (500 mg/kg) and low (50 mg/kg) oral doses of [14C]2-EH, following repeated oral dosing with unlabelled 2-EH at the low level, following dermal exposure for 6 h with a 1 g/kg applied dose of [14C]2-EH, and following a 1 mg/kg i.v. dose of [14C]2-EH. 2. The high, low and repeated low oral dose studies with 2-EH showed similar excretion balance profiles of [14C], with some evidence of metabolic saturation at the high dose. 3. No evidence of metabolic induction was seen following the repeated low oral dosing. 4. All of the oral doses were eliminated rapidly, predominantly in the urine during the first 24 h following dosing. 5. The dermal dosing resulted in only about 5% absorption of the 1 g/kg dose, with the major portion of the dose recovered unabsorbed from the dermal exposure cell at 6 h. 6. Urinary metabolites eliminated following the oral and dermal doses were predominantly glucuronides of oxidized metabolites of 2-EH, including glucuronides of 2-ethyladipic acid, 2-ethylhexanoic acid, 5-hydroxy-2-ethylhexanoic acid and 6-hydroxy-2-ethylhexanoic acid.

The metabolism and disposition of ethyl 3-ethoxypropionate in the rat.

1. The metabolism and disposition of ethyl 3-ethoxypropionate (EEP) in male Sprague-Dawley rats was studied following single oral gavage at 150 or 1500 mg/kg. The 14C-EEP was rapidly absorbed at both dose levels, and was excreted predominantly as metabolites in the urine within 24 h of administration. 2. The major urinary metabolites of EEP were monoethyl malonate and 3-ethoxypropionate. Other metabolites included malonic acid and the glycine conjugate of 3-ethoxypropionate. Trace amounts of 14C-EEP were detected after both doses. 3. 14CO2 in the expired air accounted for 34% and 20% of the dose at 150 and 1500 mg 14C-EEP/kg, respectively, mostly in the first 24 h following administration. The appearance of 14CO2 indicates extensive oxidation of the molecule, and the lower percentage of 14CO2 at the high dose possibly indicates saturation of an oxidative metabolic pathway. 4. No evidence was found for alkoxyacetic acid metabolites, such as those produced by metabolism of some low molecular weight ethylene glycol ethers.