Abstract
A new method has been developed for the quantitation of methylated blood proteins in persons exposed to methylating agents. Using different derivatizing agents, 9-fluorenylmethyl chloroformate was the most suitable with regard to both sensitivity and peak separation after subsequent HPLC. After optimization of the chromatographic conditions, two groups who worked with methyl bromide and a control group of non exposed persons were compared with regard to their adduct rates (S-methylcysteine) in the two blood proteins albumin and globin. Albumin and globin were isolated from whole blood samples. After total hydrolysis of the proteins with hydrochloric acid, an amino acid analysis was performed using HPLC with precolumn fluorescent derivatization. This method enabled a quantitation of methylated cysteine in the fmol range. Background levels of S-methylcysteine (15 nmol/g protein) in both albumin and globin were found in non-exposed control persons. Levels were up to 10 fold higher in exposed fumigators working with the methylating agent methyl bromide.
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