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Exposure to methylene chloride. III. Metabolism of 14C-labelled methylene chloride in rat.

The accumulation of methylene chloride and its metabolites in different organs and tissues was studied in an experimental series comprising 10 rats. Each rat was exposed for 1 h to radioactive methylene chloride (14C) in a concentration amounting to 1,935 mg/m3 in inspiratory air. Radioactive carbon atoms were found in the isolated carbon monoxide after exposure. There was also a close correlation between the activity of carbon monoxide extracted from the blood sample and the amount of carboxyhemoglobin (COHb) in the specimen. The largest concentration of methylene chloride and its metabolites per gram of tissue was found in white adipose tissue. This concentration had declined by more than 90% 2 h after exposure, whereas the concentration in the liver declined by about 25% during the same period. The amount accumulated in the brain displayed a decline of about 75% 2 h after exposure. Thus the examination showed that the increased concentration of COHb in the blood during exposure to methylene chloride is due to the metabolism of methylene chloride into carbon monoxide.

Adipose Tissue

Effects in the liver of methylene chloride inhaled alone and with ethyl alcohol.

When high concentrations of methylene chloride in combination with high concentrations of ethanol were inhaled for one day by guinea pigs, the extent of the hepatic damage induced suggested an antagonism between the effects of the two agents. However, exposure for five days to approximately 500 ppm of methylene chloride plus high concentrations of ethanol suggested ethanol may potentiate the effects of methylene chloride.

Animals

Exposure to methylene chloride. I Its concentration in alveolar air and blood during rest and exercise and its metabolism.

Fourteen subjects were exposed to about 870 and 1,740 mg/m3 of methylene chloride in the air during rest and physical exercise on a bicycle ergometer. The duration of each exposure period was 30 min. Each subject was exposed during four periods. The concentration of methylene chloride in the alveolar air increased in the beginning but had a tendency to level off at the end of each period. There was a high correlation between the alveolar and arterial concentration of methylene chloride. The uptake of methylene chloride was about 55 % of the supplied amount at rest, about 40% at a work load of 50 W, and about 30 and 35 % at 100 and 150 W, respectively. The concentration of carboxyhemoglobin (COHb) increased both during and after exposure. With exposure to 1,740 mg/m3 a concentration of COHb in the blood of about 0.85 g/100 ml was reached. This value corresponds to about 5.5 % COHb.

Adult

Exposure to methylene chloride. Content in subcutaneous adipose tissue.

The fat content of the body was calculated in 12 healthy male subjects aged 21 to 35 years by means of hydrostatic weighing and anthropometric estimation of skeletal weight. The subjects were exposed to a concentration of 2,600 mg of methylene chloride per cubic meter of inspired air (750 ppm) for 1 h while performing work at an intensity of 50 W on a bicycle ergometer. The uptake in the organism was measured continuously with the Douglas bag technique. The amount of methylene chloride absorbed correlated highly with degree of obesity and body weight. Needle biopsy specimens of subcutaneous adipose tissue were taken from the buttocks before exposure and 0, 1, 2, 3 and 4 h after exposure. The mean yield of tissue from the 72 biopsies was 25 mg. The concentration of methylene chloride in the adipose tissue was determined by gas chromatography, using a headspace method. The mean concentration was 10.2 mg/kg 1 h after exposure and 8.4 mg/kg after 4 h. There was a wide distribution around the mean values. In the six slim subjects the concentration in the adipose tissue during the 4 h after exposure was on an average twice that of the six more obese subjects. On the other hand, in spite of lower concentrations, the obese subjects had a greater calculated amount of methylene chloride in the total fat depots of the body. Two subjects were studied about 22 h after exposure, the concentration in subcutaneous adipose tissue being 1.6 and 1.7 mg/kg, respectively, at that time.

Adipose Tissue

Epidemiologic investigation of employees chronically exposed to methylene chloride. Mortality analysis.

A large male employee population with continuous low level work exposures to methylene chloride for up to 30 years was examined for mortality experience using non-concurrent prospective and proportionate mortality techniques. Mortality of the study group was consistent with industrial controls and less than the New York State general population. Life table analysis of the 1964 methylene chloride exposed cohort was followed through 1976 and demonstrated age group specific probabilities or 13 year survivorship equal to or better than that of three control male populations: Kodak Park, New York State, and United States.

Adolescent

Exposure to methylene chloride. II. Psychological functions.

The effect of the solvent methylene chloride on psychological functions such as reaction time, short-term memory, and numerical ability was studied in 14 healthy male subjects. Each subject was tested repeatedly during exposure to 870, 1,740, 2,600, and 3,470 mg/m3 of methylene choride in inspiratory air and during control conditions. Samples of sub ject alveolar air were taken every third minute during the experiments. No statistically significant impairement in the performance of examined functions could be observed during exposure to methylene choride as compared to control conditions. Inexposure to the highest concentration a greater irregularity of response was obtained for subject reaction time than under control conditions.

Adult

A comparative study of the effects of carbon monoxide and methylene chloride on human performance.

The purpose of this investigation was to determine whether similar carboxyhemoglobin (COHb) values obtained from inhaling carbon monoxide or methylene chloride have the same behavioral effects. The effects were assessed by the performance of 12 humans on a visual-manual, dual-task, and an auditory vigilance task. The results indicated that both substances in concentrations sufficient to produce 5-percent COHb significantly impaired human performance under difficult or demanding task conditions. The conclusion was that carbon monoxide, the main metabolite of methylene chloride, was responsible for the observed performance decrements.

Adolescent

Whole-body autoradiography and allied tracer techniques in distribution and elimination studies of some organic solvents: benzene, toluene, xylene, styrene, methylene chloride, chloroform, carbon tetrachloride and trichloroethylene.

Whole-body autoradiography has been used to study the distribution and fate of 14C-labeled benzene, toluene, xylene, styrene, methylene chloride, chloroform, carbon tetrachloride and trichloroethylene in mice after inhalational exposure. Total radioactivity, including the volatile part of the solvents were registered by autoradiography of dried, evaporated tape-fastened sections. In addition, dried, evaporated tape-fastened sections were extracted with water, trichloroacetic acid and a series of organic solvents and autoradiographed to register organs where metabolites were firmly bound. In another facet of the study, the quantitative elimination of the solvents and their metabolites via various routes was investigated. The expiration of unchanged solvents was subject to mathematical and computerized compartmental pharmacokinetic analysis. All solvents were rapidly taken up in fatty and nervous tissues where they were retained for different periods of time largely dependent on their fat solubility. Unexpectedly long retention times were noted for the chlorinated solvents, particularly for chloroforn, which showed a specific long-term retention in the cerebellum, meninges and spinal nerves, indicating interactions with specific nervous tissue receptors. Solvent metabolites appeared rapidly in several organs, mainly the liver and kidney, and were excreted both via urine and bile. Toluene and xylene metabolites were completely extractable whereas firmly bound metabolites were registered after inhalation of benzene, styrene and all chlorinated solvents, notably in the liver and kidney but also in the bronchi of the lung. Three-compartment pharmacokinetics were observed for the majority of the solvents selected for study although a two-compartment model was found to fit the elimination rate curves for styrene, methylene chloride and chloroform in the exhaled air. The fat solubility of the solvents was found to influence their pharmacokinetics since lower rate constants were generally obtained for the expiration of more fat soluble solvents.

Adipose Tissue

Determination of methylene chloride, ethylene dichloride, and trichloroethylene as solvent residues in spice oleoresins, using vacuum distillation and electron capture gas chromatography.

A quantitative gas chromatographic (GC) method is described for the determination of residual methylene chloride, ethylene dichloride, and trichloroethylene in spice oleoresins. The proposed method involves vacuum distillation in a closed system with toluene as a carrier solvent. Quantitation by electron capture GC on Porapak Q is facilitated by water extraction and by the addition of trans-1,2-dichloroethylene as an internal standard. Recoveries from oleoresins spiked at 30, 15, and 6 ppm ranged from 93 to 102%. To assess the possibility of interference from spice volatiles, the procedure was applied to 17 different spice oleoresins from 3 different manufacturers. No interferences were found, but methylene chloride levels up to 83 ppm and ethylene dichloride levels up to 23 ppm were detected. Trichloroethylene was not detected in any of the oleoresins.

Chromatography, Gas

Use of breath analysis to monitor methylene chloride exposure.

Twenty male and female subjects were exposed repetitively to methylene chloride (CH2Cl2) vapor, 50, 100, 250, and 500 ppm, for 1,3, or 7.5 h in a controlled-environment chamber. Postexposure alveolar breath samples were collected in small glass breath tubes and analyzed for CH2Cl2 by gas chromatography. From these data is a series of breath CH2Cl2 excretion curves were constructed that can be used to estimate the magnitude of a recent exposure. The CH2CL2 breath concentration in the immediate postexposure period accurately reflected the vapor concentration to which the subjects had been exposed most recently. Breath samples collected 1--2 h following exposure were accurate indicators of the time-weighted average vapor exposure experienced by the subjects during the previous 8 h of occupational exposure. Breath analysis offers a practical, noninvasive method for monitoring occupational exposure to CH2Cl2.

Adult

Effects of methylene chloride, trichloroethane, trichloroethylene, tetrachloroethylene and toluene on the development of chick embryos.

Toluene and 5 aliphatic chlorinated hydrocarbons of wide industrial use were injected into the air space of fertilized chicken eggs at 2, 3 and 6 days of incubation. The embryotoxicity was evaluated as survival and death incidences after 14 days of incubation, and also the weights and lengths of the embryos were recorded. The approximate LD50 value for trichloroethylene and trichloroethanes varied between 50 and 100 mumol/egg while for toluene, tetrachloroethylene and methylene chloride it was over 100 mumol/egg. Macroscopic malformations of various kinds were produced with doses of 5-100 mumol/egg. The teratogenic potential of the tested compounds decreased in the following order: 1,1,1-trichloroethane greater than trichloroethylene greater than methylene chloride, tetrachloroethylene, 1,1,2-trichloroethane greater than toluene greater than olive oil control.

Abnormalities, Drug-Induced

Gas chromatographic determination of residual methylene chloride and trichloroethylene in decaffeinated instant and ground coffee with electrolytic conductivity and electron capture detection.

A method is described for the quantitative determination of residual methylene chloride (MC) and trichloroethylene (TCE) in decaffeinated instant and ground roasted coffees. The residual solvents were isolated by a closed system vacuum distillation technique with toluene as a carrier solvent, chromatographed on Chromosorb 102, detected by both electron capture and electrolytic conductivity detectors, and quantitated by comparison with an internal standard. Average recoveries of MC from instant and ground coffees spiked at 1, 10, and 25 ppm were 100.0 (88-113), 93.2 (92-95), and 97.7% (94-102%); and for TCE, 97.2 (92-101), 96.2 (95-99), and 96.5% (92-100%), respectively. The results from both detectors are compared. At lower attenuations, levels less than 1 ppm can be readily measured. The procedure developed was applied to domestic and imported coffee samples.

Chromatography, Gas