六氢邻苯二甲酸酐和甲基六氢邻苯二甲酸酐所致人体血红蛋白加合物的定量方法

Quantification method of human hemoglobin adducts from hexahydrophthalic anhydride and methylhexahydrophthalic anhydride.

作者信息

Lindh C H, Jönsson B A

机构信息

Department of Occupational and Environmental Medicine, Institute of Laboratory Medicine, University Hospital, Lund, Sweden.

出版信息

J Chromatogr B Biomed Sci Appl. 1998 Jun 12;710(1-2):81-90. doi: 10.1016/s0378-4347(98)00121-2.

DOI:10.1016/s0378-4347(98)00121-2

PMID:9686874

Abstract

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.

摘要

开发了一种用于测定六氢邻苯二甲酸酐（HHPA）和甲基六氢邻苯二甲酸酐（MHHPA）与人血红蛋白（Hb）加合物的方法。该程序包括红细胞裂解、Hb溶液透析，然后进行酸水解。使用液-液和固相萃取相结合的程序纯化释放出的六氢邻苯二甲酸（HHP）和甲基六氢邻苯二甲酸（MHHP），随后用五氟苄基溴进行衍生化。使用气相色谱-质谱联用仪在以氨作为调节气体的负离子化学电离模式下分析衍生物。使用氘标记的HHP和MHHP酸作为内标。HHP酸的检测限为0.3 pmol/g Hb，MHHP酸的检测限为0.9 pmol/g Hb。HHP酸的日间精密度在2 pmol/g Hb时为18%，在13 pmol/g Hb时为10%。对于MHHP酸，精密度在2 pmol/g Hb时为27%，在22 pmol/g Hb时为14%。该方法适用于分析职业接触HHPA和MHHPA的工人的Hb加合物。