高效液相色谱-质谱联用技术同时测定人体尿液中11种代谢物

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

采用高效液相色谱-质谱联用技术定量检测人体尿样中丙烯醛、苯、1,3-丁二烯、苯乙烯、巴豆醛的巯基尿酸类代谢物及氰化氢代谢物。将解冻后的尿样经高速离心后,取100μL上清液,加入50μL内标溶液及850μL水后上样分析。色谱分析采用XSELECT HSS T3 C18色谱柱(150 mm×2.1 mm i.d,2.5μm),以乙腈-15 mmol醋酸铵溶液梯度洗脱,流速250μL/min,柱温40℃,在多反应监测模式(MRM)下采用负离子扫描模式进行定量分析。本方法共检测了16名吸烟者和6名非吸烟者的尿样,发现日吸烟量大于25支的吸烟者尿样中的烟气成分代谢物浓度高于非吸烟者。

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 μL of the supematant and 50 μL of internal standard were added and then diluted by 850 μL of water so as to be analyzed by LC-MS/MS. The compounds were separated on a XSELECT HSS 33 column using acetonitrile as mobile phase A and 15 mmol/L ammonium acetate as mobile phase B at a flow rate of 250 μL/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 μg/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 non- smokers. 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.