Dual-label high-performance liquid chromatographic assay for femtomole levels of benzo[a]pyrene metabolites.
A dual-label HPLC assay to measure femtomole quantities of ethyl acetate-extractable [3H]benzo[a]pyrene metabolites was developed. 14C-labeled metabolites of benzo[a]pyrene formed by rat liver 9000g supernatant were used as both internal standards and chromatographic markers. The percentage deviation between assays was determined to be between 11 and 13% for 9,10-dihydro-9,10-dihydroxybenzo[a]pyrene, 7,8-dihydro-7,8-dihydroxybenzo[a]pyrene, benzo[a]pyrene-3,6-quinone, benzo[a]pyrene-1,6-quinone, and 9-hydroxybenzo[a]pyrene, 22% for 4,5-dihydro-4,5-dihydroxybenzo[a]pyrene, and less than 5% for 3-hydroxybenzo[a]pyrene. The detection limit of this assay was between 3 and 10 fmol per metabolite. The application of this technique to the metabolism of [3H]benzo[a]pyrene by microsomes of hamster and human oral cavity tissue is described.
D000818:Animals; D001564:Benzo(a)pyrene / Q000378:metabolism*; D001580:Benzopyrenes / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006224:Cricetinae; D006801:Humans; D008647:Mesocricetus; D008832:Microchemistry; D008861:Microsomes / Q000378:metabolism; D008862:Microsomes, Liver / Q000378:metabolism*; D009061:Mouth Mucosa / Q000378:metabolism*
D001580:Benzopyrenes; D001564:Benzo(a)pyrene