QUANTIFICATION METHOD OF HUMAN HEMOGLOBIN ADDUCTS FROM HEXAHYDROPHTHALIC ANHYDRIDE AND METHYLHEXAHYDROPHTHALIC ANHYDRIDE

A method was developed for the determination of human hemoglobin (Hb) adducts from hexahydrophthalic anhydride (HHPA) and methylhexahydropht halic anhydride (MHHPA). The procedure includes lysis of erythrocytes, dialysis of the Hb-solution followed by acid hydrolysis. The released hexahydrophthalic (HHP) acid and methylhexahydrophthalic (MHHP) acid were purified using a combined liquid-liquid and solid-phase extractio n procedure followed by derivatization with pentafluorobenzyl bromide. The derivatives were analyzed using GC-MS in negative ion chemical io nization mode with ammonia as moderating gas. As internal standards, d euterium-labeled. HHP and MHHP acids were used. The detection limits w ere 0.3 pmol/g Hb for HHP acid and 0.9 pmol/g Hb for MHHP acid. ?he be tween-day precisions for HHP acid were 18% at 2 pmol/g Hb and 10% at 1 3 pmol/g Hb. For MHHP acid, the precision was 27% at 2 pmol/g Hb and 1 4% at 22 pmol/g Hb. The method was applicable for analysis of Hb adduc ts from workers occupationally exposed to HHPA and MHHPA. (C) 1998 Els evier Science BN. All rights reserved.