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 methylhexahydrophthalic 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 extraction procedure followed by derivatization with pentafluorobenzyl bromide. The derivatives were analyzed using GC-MS in negative ion chemical ionization mode with ammonia as moderating gas. As internal standards, deuterium-labeled HHP and MHHP acids were used. The detection limits were 0.3 pmol/g Hb for HHP acid and 0.9 pmol/g Hb for MHHP acid. The between-day precisions for HHP acid were 18% at 2 pmol/g Hb and 10% at 13 pmol/g Hb. For MHHP acid, the precision was 27% at 2 pmol/g Hb and 14% at 22 pmol/g Hb. The method was applicable for analysis of Hb adducts from workers occupationally exposed to HHPA and MHHPA.
Department of Occupational and Environmental Medicine, Institute of Laboratory Medicine, University Hospital, Lund, Sweden.
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Lindh
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Jönsson
D002138:Calibration; D003956:Dialysis; D004784:Environmental Monitoring; D004853:Epoxy Resins / Q000032:analysis*; D004912:Erythrocytes / Q000737:chemistry; D005069:Evaluation Studies as Topic; D008401:Gas Chromatography-Mass Spectrometry; D006454:Hemoglobins / Q000032:analysis*; D006801:Humans; D006868:Hydrolysis; D016273:Occupational Exposure; D010796:Phthalic Anhydrides / Q000032:analysis*; D012680:Sensitivity and Specificity
D004853:Epoxy Resins; D006454:Hemoglobins; D010796:Phthalic Anhydrides; C005752:araldite