Analysis of styrene oxide-globin adducts based upon reaction with Raney nickel.
Ting-D; Smith-MT; Doane-Setzer-P; Rappaport-SM
Carcinogenesis 1990 May; 11(5):755-760
A method was described for the determination of styrene-oxide/globin adducts. Male Sprague-Dawley-rats were given a single intraperitoneal injection of styrene (100425) in corn-oil at 0.5, 1, 2, and 3 millimoles/kilogram. Blood was obtained 20 hours after dosing. The analysis technique used takes advantage of the reaction between alkylated globin and Raney nickel which cleaves the carbon to sulfur bond in the styrene-oxide/cysteine adduct to form 1- phenylethanol (1-PE) and 2-phenylethanol (2-PE). These alcohols were reacted next with pentafluorobenzoyl-chloride and analyzed by gas chromatography/electron capture detection. The detection limit of the method was 0.04 nanomole adducts/sample, indicating that it should be possible to detect adducts in the blood of people who are occupationally exposed to at least 18mg/m3 of styrene. Analysis of globin from human whole blood which had been modified with carbon-14 labeled styrene-oxide (96093) indicated that 6% of the total globin adducts were detected. The method was then applied to human and rat blood which had been treated with styrene-oxide in-vitro and to blood from rats given a single intraperitoneal dose of styrene in- vivo. Results indicated that 77 times more adducts were detected at a given dose from rat globin than from human globin and that only blood of rats. The reaction with Raney nickel was conducted at 5 degrees-C to minimize unfavorable side reactions. The optimal amount of Raney nickel was 5 to 6 grams/gram globin. Since the recovery of 1-PE was not reproducible, only 2-PE was used for quantitation.
NIOSH-Publication; NIOSH-Grant; Laboratory-animals; Chemical-reactions; Analytical-methods; Chromatographic-analysis; Biological-monitoring; Genotoxic-effects; Blood-analysis
Biomedical & Environ Hlth Scis University of California School of Public Health Berkeley, CA 94720
University of California Berkeley, Berkeley, California