Determination of 1-hydroxypyrene in human urine by high-performance liquid chromatography with fluorescence detection using a deuterated internal standard.
A high-performance liquid chromatographic method has been developed for the quantification of 1-hydroxypyrene (1-OHP) in human urine using deuterated 1-hydroxypyrene ([2H9]1-OHP) as an internal standard with fluorescence detection. [2H9]1-OHP was prepared enzymatically from deuterated pyrene ([2H10]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 microg/L and the calibration range was from 1 to 100 nmol/L. Urine sample treatment involved enzymatic hydrolysis followed by solid-phase extraction using Sep-Pak C18 cartridges. The proposed method was used to determine urinary 1-OHP in smokers and non-smokers.
Graduate School of Natural Science and Technology, Kanazawa University, Ishikawa, Japan.
Thaneeya
T
Chetiyanukornkul
Akira
A
Toriba
Ryoichi
R
Kizu
Tsunehisa
T
Makino
Hiroyuki
H
Nakazawa
Kazuichi
K
Hayakaw
D000328:Adult; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003903:Deuterium; D006801:Humans; D011721:Pyrenes / Q000032:analysis*; D012015:Reference Standards; D012907:Smoking / Q000652:urine; D013050:Spectrometry, Fluorescence / Q000379:methods*
D011721:Pyrenes; D003903:Deuterium; C033146:1-hydroxypyrene