Inhalation teratology and reproduction studies with 1,1-dichloro-2,2,2-trifluoroethane (HCFC-123).

HCFC 123 is one of the chemicals being developed as a replacement for CFC 11 in refrigerant and solvent applications. Supplementing earlier rat teratology studies, a rabbit inhalation teratology study was conducted. In addition, one-generation and two-generation inhalation reproduction studies were conducted. In the teratology study, the pregnant rabbits were exposed to levels of 0 (control), 500, 1500, and 5000 ppm, 6 hr per day from Days 6 through 18 of gestation. Slight body weight losses and reduced food consumption were seen in does in all three exposure level groups. This response followed an exposure-related pattern. There were no other signs of maternal toxicity. There was also no evidence of treatment-related effects on the kits. A probe one-generation reproduction study was conducted. In this study four groups of 12 male and 12 female rats were exposed to vapors of HCFC 123 6 hr per day, 7 days per week from 4 weeks prior to mating through weaning of their offspring. The exposure levels for this study were 0 (control), 300, 1000, and 5000 ppm. There were no effects on mating and fertility, or on pup survival or birth weight. A two-generation study was subsequently conducted. In this study, five groups of 32 male and female rats were exposed to HCFC 123 from 6 weeks of age through weaning. From the offspring of these animals, groups of 28 males and females were selected for the F1 generation. These animals were exposed to HCFC 123 from weaning (4 weeks of age) through weaning of the F1 generation. All exposures were 6 hr per day, 7 days per week. The exposure levels for this study were 0 (control), 30, 100, 300, and 1000 ppm. There were no effects on any of the fertility or reproductive indices measured. As with prior studies, decreases in serum triglyceride levels were seen. Pup survival and birth weight were unaffected by treatment. Pup body weight gain was lower in all treatment groups during nursing, following an exposure-related pattern. Since weight gain for the F1 animals was normal following weaning, this depression of body weight gain may be related to the depression of serum triglycerides. In addition, liver weights of the adult rats exposed to levels of 100 ppm and higher of HCFC 123 were higher than controls, histological examination revealed only hepatic enlargement and vacuolation. It was concluded that exposure to HCFC 123 did not cause reproductive effects although it did effect the body weight gain of the offspring during lactation.

Acute, subchronic, and developmental toxicity and genotoxicity of 1,1,1-trifluoroethane (HFC-143a).

The toxicity potential of 1,1,1-trifluoroethane (HFC-143a), a CFC alternative, was evaluated in several acute, subchronic, and developmental toxicity studies by the inhalation route and in genotoxicity studies. HFC-143a has a very low acute inhalation toxicity potential as shown by a 4-hr LC50 of > 540,000 ppm in rats. HFC-143A has a low potential to induce cardiac sensitization in experimental screening studies in dogs; only the highest concentration tested--300,000 ppm--elicited a cardiac sensitization response. In an initial 4-week nose-only inhalation study, male and female rats were exposed 6 hr/day, 5 days/week at concentrations of 0, 2000, 10,000, or 40,000 ppm. Females showed no evidence of toxicity at any exposure level; male rats did exhibit degenerative changes only in the tests at all exposure levels. However, because of exposure system irregularities, which resulted in excessive temperature conditions and stress in the HFC-143a-exposed groups, the study was repeated in male rats exposed by whole-body inhalation. In this repeat study no toxicity was observed at < or = 40,000 ppm. Moreover, a subsequent 90-day whole-body inhalation study in rats exposed 6 hr/day, 5 days/week at 0, 2000, 10,000, or 40,000 ppm resulted in no evidence of toxicity at any exposure concentration. The results of the second 4-week and the 90-day studies using whole-body exposures indicate that the findings from the first 4-week study were related to the stress induced by excessive temperatures and nose-only restraint. Therefore, the no-observed-effect level (NOEL) for rats repeatedly exposed up to 90 days was considered to be 40,000 ppm. In developmental toxicity studies with rats and rabbits, an increase in visceral variations or skeletal malformations was observed, respectively, at HFC-143a concentrations of 2000, 10,000 or 40,000 ppm (rat) or at the low and high concentrations (rabbit). Because of the unusually low control incidence of variations (1.6% per litter in the control versus 6.8-16.8% for historical control values), the lack of a clear dose-response relationship, and the lack of other developmental effects, these findings were not considered related to HFC-143a exposure. In addition, results from genotoxicity studies (Ames, chromosomal aberration with human lymphocytes, mouse micronucleus) demonstrated that HFC-143a was not mutagenic.

Acute and subchronic toxicity of 1,1-dichloro-1-fluoroethane (HCFC-141b).

The acute and subchronic toxicity of 1,1-dichloro-1-fluoroethane (HCFC-141b), a CFC alternative, was evaluated in several acute and subchronic studies to assist in establishing proper handling guides. Data from acute toxicity studies in rats and rabbits demonstrated that HCFC-141b has very low acute toxicity. HCFC-141b was not a skin irritant, but was a mild eye irritant, in rabbits and was not a skin sensitizer in guinea pigs. Skin application of HCFC-141b to rabbits at 2000 mg/kg body weight produced no adverse effects. Oral administration at 5000 mg/kg body weight did not cause any deaths or clinical signs of toxicity in rats. The 4-hr LC50 for HCFC-141b was about 62,000 ppm in rats. Repeated exposures of rats for 6 hr/day, 5 days/wk for up to 90 days at concentrations of 2000, 8000 or 20,000 ppm did not result in significant adverse effects. Minor, but dose-dependent, reductions in body weight were observed in male and female rats during the 90-day study. Decreased responsiveness was also observed in rats but only at 20,000 ppm. An increase in serum cholesterol or triglycerides was observed in male and female rats at 20,000 ppm, and in males at 8000 ppm. No specific organ pathology was noted in these subchronic inhalation studies. The no-observable-adverse-effect level (NOAEL) from these studies was 8000 ppm. Results from other studies demonstrate that HCFC-141b was not neurotoxic in rats. As with trichlorofluoroethane (CFC-11), a cardiac sensitization response to an intravenous epinephrine challenge occurred in dogs with HCFC-141b at 5000 ppm and higher concentrations in experimental screening studies.

Evaluation of the genotoxicity potential and chronic inhalation toxicity of 1,1-dichloro-1-fluoroethane (HCFC-141b).

A battery of in vitro and in vivo tests were conducted on HCFC-141b as a vapour. Bacterial gene mutation assays with Escherichia coli and Salmonella typhimurium were negative in all tester strains. In vitro chromosomal aberration assays were positive on CHO cells but negative on human lymphocytes. Moreover, HCFC-141b was negative in vivo in a mouse micronucleus inhalation assay. On the basis of these data and previously reported genotoxicity testing, HCFC-141b is considered non-genotoxic. Groups of 80 male and 80 female Sprague-Dawley rats were exposed, by inhalation (6 hr/day, 5 days/wk) to vapours of HCFC-141b for 104 wk at target concentrations of 0 (control), 1500, 5000 and 20,000 ppm (increased from 15,000 ppm after 17 wk of exposure). No exposure-related effects of toxicological significance were noted with respect to survival, clinical signs, ophthalmoscopy, haematology, clinical chemistry, urinalysis or organ weight analysis. Reduced food intake and body weight gain were noted in both sexes of the 15,000 ppm group during the first 16 wk; thereafter, body weight gains in all groups were similar although the intergroup differences in body weight remained evident. Reduced food intake persisted in both sexes through wk 52 and in females during the second year of exposure. Treatment-related effects on macroscopic pathology were confined to increased incidences of testicular masses and altered appearance. Microscopic pathology examinations confirmed the testes as the target organ with findings of increased incidences of benign interstitial cell tumours and hyperplasia at 5000 and 20,000 ppm. The no-observable-adverse-effect level (NOAEL) was 1500 ppm. The testicular changes at high exposure levels were considered to be due to a change of the senile hormonal imbalance in geriatric rats and of little significance for the assessment of human health effects.

1,1,1,2-Tetrafluoroethane: repeat exposure inhalation toxicity in the rat, developmental toxicity in the rabbit, and genotoxicity in vitro and in vivo.

Subchronic and chronic studies were carried out in the rat and a developmental toxicity study in the rabbit with exposures to 1,1,1,2-tetrafluoroethane (HFC 134a) by inhalation. In the rat repeated exposure to 50,000 ppm HFC 134a for 13, 52, and 104 weeks elicited no effect on clinical condition, growth, and survival, or on a variety of hematological, clinical chemistry, and urinary parameters. Treatment-related pathological changes were seen only at study termination at 2 years and were confined to increased incidence of Leydig cell hyperplasia and adenoma in male rats exposed to 50,000 ppm. The tumors, which were also seen in control animals, were benign and not life-threatening. A battery of in vitro and in vivo tests gave no evidence of genotoxic activity. With exposure to pregnant rabbits, the only treatment-related effects were of minimal maternal toxicity at high exposure concentrations; there were no effects on fetal development. It is concluded that HFC 134a is of very low toxicity and should be an acceptable alternative to CFCs.

Inhalation teratology and two-generation reproduction studies with 1,1-dichloro-1-fluoroethane (HCFC-141b).

HCFC-141b is one of the chemicals being considered as a replacement for CFC 11 in solvent and foam-blowing applications. Teratology studies were conducted in both rats and rabbits and a two-generation reproduction inhalation toxicity study was conducted in rats. The pregnant rabbits were exposed to levels of 0 (control), 1400, 4200 and 12,600 ppm HCFC-141b from day 7 to day 19 of gestation (6 hr/day). There was no evidence of developmental or teratogenic effects on the foetuses. The pregnant rats in the teratology study were exposed to levels of 0 (control), 3200, 8000 and 20,000 ppm from days 6 to 15 of gestation (6 hr/day). In the 20,000 ppm exposure group, there was an increase in implantation losses; furthermore, in this group, foetal weights tended to be lower than controls. As with the rabbits, there was no evidence of a teratogenic effect. The reproduction study was conducted at exposure levels of 0, 2000, 8000 and 20,000 ppm, 7 days/wk starting approximately 10 wk before the first pairing. Adult rats exposed at 20,000 ppm (and, to a lesser extent, those exposed to 8000 ppm) showed increases in water intake, slight increases in food consumption, and decreases in body weight. Following the mating of the F0 parents, there were fewer litters in the 20,000 ppm exposure level group than in controls. When these parents were then paired with different partners, again, the number of litters was lower in the 20,000 ppm group, although most of the animals that did not produce litters the first time mated successfully the second time. When the F1 animals were mated to produce the second generation, the number of litters was comparable for all groups. In the second F0 mating and the F1 mating, the number of pups per litter was lower at 20,000 ppm; although birth weights were comparable, body weight gain tended to be slower in the high-level exposure group. Survival was good in all groups. At 8000 ppm no significant effects were observed in the pups and only minimal signs in the adults. The 2000 ppm exposure level represented a clear no-observed-effect level for all indices.

Evaluation of the neurotoxic potential of chemicals in animals.

This review provides a scientific view on how to evaluate effectively the neurotoxic potential of chemicals in order to provide adequate safeguards for human health. Detection of compounds that may cause direct, persistent, adverse effects on the nervous system should be given the most critical attention. Evaluation of the neurotoxic potential of a chemical should include descriptions of functional and morphological effects as well as the determination of the dose response, no-observed-effect level, time course and reversibility of effects. Evaluation of the nervous system in the context of standard toxicity studies that use a variety of species and study durations are appropriate screening tests (Tier 1) for the detection of potential neurotoxicity. Studies specifically designed to assess neurotoxicity (Tier 2) should be performed with chemicals for which there is an indication of neurotoxic potential and where the available data are inadequate for risk assessment. Tier 2 studies should evaluate the function and structure of the nervous system by comprehensive clinical examinations and neuropathological assessment. These studies may be conducted in conjunction with standard toxicity studies so that any potential neurotoxicity can be interpreted in the context of other systemic toxicity. More specific neurotoxicity tests (Tier 3) may be necessary for advanced characterization of discovered neurotoxicants.

On the metabolism of clofibride, a hypolipaemic drug.

The absorption and metabolism of clofibride, a new hypolipaemic drug of p-chlorophenoxyisobutyric type, were investigated in the CD rat, the beagle and the olive baboon monkey. Clofibride is rapidly and massively resorbed and hydrolysed into 4-chlorophenoxyisobutyric acid (CPIB) and 4-hydroxy-N-dimethylbutyramide (HMB). CPIB, in free and glucuroconjugated form, and its metabolite, 4-chlorophenol, in the form of the glucuronate ether, are found in the serum of the rat. HMB is rapidly metabolized. The half-life of CPIB, the main active metabolite, in the serum is about 12 h in the rat, 43 +/- 9 h in the dog and 6 +/- 1 h in the baboon. In the rat, peak hypocholesterolaemic activity occurs late--24 h after administration of the drug and 20 h after peak concentration of CPIB in the blood. The half-life of 4-chlorophenol glucuronate ether in the serum is about 4 h whereas that of HMB is about 3 h. In the rat, the elimination of clofibride takes place mainly via the urine since 70% of the dose administered is found in the form of free or conjugated CPIB, 10% in the form of HMB or one of its metabolites, in 48 h samples of urine. Over the same period, faecal elimination accounts for no more than 2% of the dose ingested. In addition, in this species, the CPIB, 30% of which is secreted via the biliary route without being eliminated in the faeces, undergoes an enterohepatic circulation.