INRA, UMR 1089 Xénobiotiques INRA-ENVT, 180 chemin de Tournefeuille, BP 93173, Toulouse, 31027, France.
Anal Bioanal Chem. 2010 Mar;396(5):1691-701. doi: 10.1007/s00216-009-3389-1. Epub 2010 Jan 18.
An original method was developed to separate, identify and quantify the different benzo(a)pyrene (B(a)P) metabolites formed through oxidative and conjugative pathways. All B(a)P metabolites were separated by an improved high-performance liquid chromatography method, then detected and quantified relatively by online radioactivity detection. At the same time, metabolite structures were characterised by tandem mass spectrometry using two complementary ionisation modes: electrospray ionisation in the negative mode and atmospheric pressure chemical ionisation in the positive mode. This method was successfully applied to the analysis of B(a)P metabolites, produced by incubation of B(a)P with the ex vivo pig ear skin model. These include glucuronic acid and sulphate conjugates of B(a)P-OHs and B(a)P-diols, as well as direct phase I metabolites: B(a)P-tetrol, B(a)P-diones, B(a)P-catechols, B(a)P-diols and B(a)P-OHs.
开发了一种原始方法来分离、鉴定和定量通过氧化和共轭途径形成的不同苯并(a)芘（B(a)P）代谢物。所有 B(a)P 代谢物均通过改进的高效液相色谱法分离，然后通过在线放射性检测相对检测和定量。同时，使用两种互补的离子化模式（负离子模式下的电喷雾离子化和正离子模式下的大气压化学离子化）通过串联质谱对代谢物结构进行表征。该方法成功应用于 B(a)P 与离体猪耳皮肤模型孵育产生的 B(a)P 代谢物的分析。这些包括 B(a)P-OH 和 B(a)P-二醇的葡萄糖醛酸和硫酸盐缀合物，以及直接的 I 相代谢物：B(a)P-四醇、B(a)P-二酮、B(a)P-儿茶酚、B(a)P-二醇和 B(a)P-OH。
