A new sensitvie fluorometric assay for the metabolism of (–)-7,8-dihydroxy-7,8-dihydrobenzo[α]pyrene by human hair follicles

Abstract
A new sensitive fluorometric assay was established to measure the stereospecific formation of benzo [α]pyrene tetrols formed after cytochrome P450-dependent metabolism of (−)-7,8-dihydroxy-7,8-dihydrobenzo by human hair follicles. This simple assay requires three human hair follicles and a low (0.5−2.0 μM) substrate concentration and has a limit of detection of ∼0.3 fmol of tetrols. Freshly isolated human hair follicles from 20 adult volunteers (10 non-smokers and 10 smokers) were assayed. While intersubject and seasonal variations were observed, the assay was found to be reproducible for a given subject. This rapid and non-invasive assay provides a new means for metabolic phenotyping of human subjects for their capacity to metabolize (−)-7,8-dihydroxy-7,8-dihydrobenzo [α]pyrene to its carcino genic form (+)anti-benzo[α] diolepoxide.

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