INVESTIGATION ON TOXICITIES OF ALTERNATIVES FOR CHLOROFLUOROCARBONS AND ITS RISK ASSESSMENT USING MOLECULAR AND BIOLOGICAL MARKERS.

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Urine was collected from the rats exposed to 1-bromopropane. One hundred μL of urine was added 400μL of saturated sodium chloride solution, and then extracted with 500μL of ethyl acetate for three times. The collected ethyl acetate layer was dehydrated with dried sodium sulfoxide, and dried with centrifuging evaporator. The dried sample was resolved with 40μL of ethylacetate and derivatized with TBDMS. The result showed the metabolite N-acetyl-S-propylcysteine was dose-dependent and available as an internal exposure marker.Blood was collected from the rats exposed to 1-bromopropane. Globin was extracted from blood samples using acetone-oxalic acid mixture. Globin was hydrolyzed with acid under nitrogen, and S-propylcysteine was identified using LC-MS/MS.The cerebrum, cerebellum, brainstem and spinal cord were taken from the rats exposed to 1-bromopropane under deep anesthesia. The tissue was homogenized and the soluble fraction was obtained. Neuro-specific protein γ-enolase and glia-specific protein β-S100 was quantified with a sandwich type enzyme immunoassay. Glutatione (GSH and GSSG) was also measured by enzymatic cycling method. The result showed that gamma-enolase was decreased specifically in the cerebrum and 1-bromopropane has a great adverse effect on the central nervous system.The testes were taken from the rats exposed to 1-bromopropane under deep anesthesia. Immunostaining with α-enolase showed clearly specific localization of α-enoalse to Sertoli cells in the testis. The quantification with enzyme immunoassay showed dose-dependent increase in alpha-enolase in the testis, suggesting the effects of 1-bromopropane on Sertoli cells.