title: Derivatization method for sensitive determination of 3-hydroxybenzo[a]pyrene in human urine by liquid chromatography-electrospray tandem mass spectrometry.
abstract: While urinary 3-hydroxybenzo[a]pyrene (3-OHBaP) can indicate exposure to carcinogenic benzo[a]pyrene (BaP) in humans, most previous analytical methods could not detect it in the general population due to a lack of sufficient sensitivity. In this study, a new liquid chromatography-electrospray mass-mass spectrometry (LC-ESI-MS/MS) method was developed to analyze 3-OHBaP by employing an optimized derivatization reaction with dansyl chloride (DNS) in the aqueous phase. The instrument detection limit (IDL) of the newly developed method was 10ng/L, which is 60-fold lower than without derivatization. The method was successfully applied to analyze 3-OHBaP and 1-hydroxypyrene (1-OHPy) in human urine combined with enzymatic hydrolysis, C18 enrichment, liquid-liquid extraction and silica cartridge cleanup. The overall method recoveries were 68.7±5.47% for 3-OHBaP and 72.3±4.96% for 1-OHPy, with matrix effects of <10%. The limits of quantification (LOQs) in urine were 0.3±0.02ng/L and 0.5±0.02ng/L (n=3), respectively. The method was applied to analyze seven urine samples of volunteers, and 3-OHBaP was detected in five samples. The concentration of urinary 3-OHBaP ranged from <LOD to 2.30ng/L, much lower than those (42.0-216ng/L) of 1-OHPy. The developed method provides a new way to sensitively determine 3-OHBaP in urine of the general population.
authors.affiliation: MOE Laboratory for Earth Surface Processes, College of Urban and Environmental Sciences, Peking University, Beijing 100871, China.
authors.forename: Kai
authors.initials: K
authors.lastname: Luo
authors.affiliation: MOE Laboratory for Earth Surface Processes, College of Urban and Environmental Sciences, Peking University, Beijing 100871, China.
authors.forename: Qun
authors.initials: Q
authors.lastname: Gao
authors.affiliation: MOE Laboratory for Earth Surface Processes, College of Urban and Environmental Sciences, Peking University, Beijing 100871, China. Electronic address: hujy@urban.pku.edu.cn.
authors.forename: Jianying
authors.initials: J
authors.lastname: Hu
mesh_terms: D001564:Benzo(a)pyrene / Q000032:analysis*; D002273:Carcinogens / Q000032:analysis; D002853:Chromatography, Liquid; D004785:Environmental Pollutants / Q000652:urine*; D006801:Humans; D057230:Limit of Detection; D011721:Pyrenes / Q000652:urine; D053719:Tandem Mass Spectrometry; D016482:Urinalysis / Q000379:methods*
chemical_list: D002273:Carcinogens; D004785:Environmental Pollutants; D011721:Pyrenes; D001564:Benzo(a)pyrene; C033146:1-hydroxypyrene
keywords: 3-Hydroxybenzo[a]pyrene; Chemical derivatization; Human urine; Internal exposure