Raha C, Hines R, Bresnick E
J Chromatogr. 1984 May 18;291:231-9. doi: 10.1016/s0021-9673(00)95024-6.
To explain the biological activity of 6-nitrobenzo[a]pyrene (6-nitroBaP), male Sprague-Dawley rats were induced with 3-methylcholanthrene. Liver microsomes were incubated with magnesium chloride, an NADPH generating system and 6-nitroBaP in acetone. The mixture was chilled under oxygen-free argon gas and protein was precipitated with an equal volume of cold methanol containing triethylamine. Protein was further precipitated with zinc and sodium sulfate and centrifuged. Both the sediment and the supernatant were extracted with benzene and ethyl acetate. The organic extract was washed with water, 2% sodium hydroxide solution, water and then dried with anhydrous sodium sulfate. Solvents were removed and the residue was chromatographed on silica gel plates with hexane containing increasing amounts of benzene. The UV and mass spectra of products were examined. Liver microsomal metabolites of 6-nitroBaP consisted of 7,8- and 9,10-dihydrodiols and also benzo[a]pyrene (BaP) and BaP-quinones. cis-Forms of 6-nitroBaP-7,8- and -9,10-dihydrodiols were synthesized.
为解释6-硝基苯并[a]芘（6-nitroBaP）的生物活性，用3-甲基胆蒽诱导雄性斯普拉格-道利大鼠。将肝微粒体与氯化镁、一个NADPH生成系统以及丙酮中的6-nitroBaP一起孵育。混合物在无氧氩气下冷却，并用等体积含三乙胺的冷甲醇沉淀蛋白质。蛋白质再用锌和硫酸钠沉淀并离心。沉淀物和上清液都用苯和乙酸乙酯萃取。有机萃取物用水、2%氢氧化钠溶液、水洗涤，然后用无水硫酸钠干燥。除去溶剂，残留物在硅胶板上用含逐渐增加量苯的己烷进行色谱分析。检测产物的紫外光谱和质谱。6-nitroBaP的肝微粒体代谢产物包括7,8-和9,10-二氢二醇，还有苯并[a]芘（BaP）和BaP-醌。合成了6-nitroBaP-7,8-和-9,10-二氢二醇的顺式异构体。
