© 1990 Oxford University Press
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Analysis of styrene oxide–globin adducts based upon reaction with Raney nickel
Department of Biomedical and Environmental Health Sciences, School of Public Health. University of California Berkeley, CA 94720, USA
1To whom correspondence should be addressed
A new method has been developed for determination of styrene oxide–globin adducts. The technique takes advantage of the reaction between alkylated globin and Raney nickel, which cleaves the carbon–sulfur bond in the styrene oxide–cysteine adduct to form 1-phenylethanol (1-PE) and 2-phenylethanol (2-PE). These alcohols are then reacted with pentafluorobenzoyl chloride and analyzed by GC–ECD. The method appears useful for biological monitoring of individuals exposed to styrene and, potentially, to other chemicals or their electrophilic metabolites which can react with cysteine residues in available proteins. The detection limit of the method, which is 0.04 ninol adducts/sample, indicates that it should be possible to detect adducts in the blood of people who are occupationally exposed to at least 18 mg/rn of styrene. Analysis of globin from human whole blood which had been modified with [14C[ oxide indicated that 6% of the total globin adducts were detected. The méthod was applied to human and rat blood which had been treated with styrene oxide in vitro and to blood from rats given a single i.p. dose of styrene in vivo. Results from these experiments indicate that 77 times more adducts were detected at a given dose from rat globin than from human globin and that only 0.015% of the styrene dose was bioavailable as styrene oxide in the blood of rats. The reaction with Raney nickel is conducted at 5°C to minimize unfavorable side reactions, such as degradation of 1- and 2-PE and conversion of styrene glycol to 1- and 2-PE. The optimal amount of Raney nickel was found to be 5–6 g/g globin. Since the recovery of 1-PE was not reproducible, only 2-PE is used for quantitation.