Ethylene oxide에 폭로에 의해 형성된 헤모글로빈 adduct의 분석에 관한 연구

A gas chromatography/electron impact mass spectrometric assay method was developed for the determination of Hb-adduct, 2-(hydroxyethyl)valine (HEVal), of ethylene oxide(EO). Globin was separated from hemoglobin by acid iso-propanol and ethyl acetate, then HEVal was isolated as PFPITH-HEVal by Edman degradation. PFPITH-HEVal was silylated with N-methyl-N-(tert-butyl-dimethylsilyl)trifluoroacetamide(MTBDMSTFA)-NH4I (1000:4) under catalysis of dithioerythritol. The detection limit of the assay was 5.8 pmol/g based upon assayed hemoglobin of 0.1g. Two groups of mice were exposed to EO for 0.5 and 1.0 hr/day, respectively at 400ppm during 4 weeks. As the result, the adduct levels increased according to the exposure time with the linearity of 0.7011 and 0.8914, respectively, HEVal was very valuable as biomarker for the exposure of EO. In human, HEVal was analyzed until 8.33 pmol/mg.