Simultaneous Determination of 11 Volatile Organic Compounds Metabolites in Human Urine Using High Performance Liquid Chromatography Coupled with Electrospray Ionization Tandem Mass Spectrometry

We developed and validated a method using high performance liquid chromatography ( HPLC) coupled with electrospray ionization tandem mass spectrometry ( ESI-MS/MS) to simultaneously quantify 11 urinary volatile organic compound ( VOC) metabolites. The frozen urinary samples were unfrozen at room temperature and centrifuged to remove settled precipitate. 100 tiLL of the supernatant and 50 mu L of internal standard were added and then diluted by 850 mu L of water so as to be analyzed by LC-MS/MS. The compounds were separated on a XSELECT HSS T3 column using acetonitrile as mobile phase A and 15 mmoUL ammonium acetate as mobile phase B at a flow rate of 250 mu L/min. The analytes were determined qualitatively and quantitatively under multi-reaction monitoring ( MRM) and negative polarity mode. The limits of detection ranged from 0.5-30 mu g/L and the spiked recoveries of the analytes ranged from 89. 8% 123. 8% with RSDs of 3. 1% 14. 4%. This method was applied to 22 h urine samples collected from 16 smokers and 6 nonsmokers. The results showed that the contents of the metabolites in smoker's urine were higher than those of nonsmokers, especially, smoking more than 25 cigarettes per day.