Abstract
A high-performance liquid chromatographic method has been developed for the quantification of 1-hydroxypyrene (1-OHP) in human urine using deuterated 1-hydroxypyrene ([ 2H 9]1-OHP) as an internal standard with fluorescence detection. [ 2H 9]1-OHP was prepared enzymatically from deuterated pyrene ([ 2H 10]Pyr) with cytochrome P450 1A1. It eluted immediately prior to non-deuterated 1-OHP on alkylamide-type reversed-phase columns and had nearly the same fluorescence characteristics as non-deuterated 1-OHP. The detection limit was 0.1 μg/L and the calibration range was from 1 to100 nmol/L. Urine sample treatment involved enzymatic hydrolysis followed by solid-phase extraction using Sep-Pak C 18 cartridges. The proposed method was used to determine urinary 1-OHP in smokers and non-smokers.
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