CORESTA Congress, Berlin, 2016, Smoke Science/Product Technology Groups, STPOST 28

Determination of benzo[a]pyrene in smokers’ urine by online gel permeation chromatography–solid phase extraction–gas chromatography/mass spectrometry

SI Xiaoxi; ZHU Ruizhi; LIU Zhihua; ZHANG Fengmei; LIU Chunbo; SHEN Qinpeng; HE Pei; MIAO Mingming; YANG Guangyu
Key Laboratory of Tobacco Chemistry of Yunnan, R&D Center of China Tobacco Yunnan Industrial Co., Ltd., Kunming, P.R. China

A new method for determining the benzo[a]pyrene (B[a]P) in samples of smokers’ urine was developed with an online gel permeation chromatography-solid phase extraction-gas chromatography/mass spectrometry (GPC-SPE-GC/MS) hyphenated system. Urine samples from smokers were extracted with cyclohexane containing D12-B[a]P as an internal standard. The extract was separated by gel permeation chromatography and the B[a]P fraction was cut into a solid phase extraction cartridge and enriched. The enriched B[a]P was eluted with acetone and determined by gas chromatography-mass spectrometry (GC-MS). The method utilised the online connections of gel permeation chromatography, solid phase extraction and gas chromatography/mass spectrometry, eliminating the need for multi-step manual purification. By incorporating gel column in liquid chromatography, the volume of sample injection increased to 200 µL and sample loading capacity was 10 times that of a conventional micro silica gel column. The purified large volume fractions from liquid chromatography were injected into the gas chromatography directly via online solid phase extraction, enhancing sensitivity. For B[a]P concentrations in the range of 0.08-10 ng/mL, the method had good linearity with the correlation coefficient of 0.9992. The intra-day and inter-day relative standard deviations (RSDs) were 3.26% and 3.87%, the limit of detection (LOD) and limit of quantification (LOQ) were 0.022 and 0.093 ng/L, respectively; and the recoveries ranged from 95.6% to 97.2% at three spiked levels. The method is automated, simple, fast, sensitive, accurate, and suitable for determining B[a]P levels in smokers’ urine.