高效液相色谱-串联质谱法测定尿中可替宁

Determination of cotinine in urine by high performance liquid chromatography-tandem mass spectrometry

目的高效液相色谱-串联质谱法(HPLC-MS-MS)测定尿中可替宁的分析方法。方法采用甲基叔丁基醚(MTBE)为萃取剂,在弱酸条件下萃取尿液中的可替宁。使用Zorbax Eclipse Plus C18(150 mm×3 mm,1.8μm)色谱柱,以0.1%甲酸水-色谱醇甲醇(含0.1%甲酸)为流动相进行梯度洗脱,流速为0.4 ml/min,柱温为45℃。使用ESI源正离子扫描模式质谱检测。结果在1 ng/ml~2 000 ng/ml浓度内,方法线性相关系数为0.999 8。检出限为0.5 ng/ml,加标回收率在93.3%~96.4%,RSD为0.86%~3.89%。结论该方法简单高效,溶剂使用量少,具有高通量、高灵敏度的特点,适用于主动吸烟和被动吸烟人群尿液可替宁含量的检测。

Objective To develop a high performance liquid chromatography-tandem mass spectrometry(HPLC-MS-MS)method for the determination of cotinine in urine.Methods Methyltert-butyl ether(MTBE)is applied as liquid extraction solvent for urine cotinine purification under acidic condition.Zorbax Eclipse Plus C18(150 mm×3 mm,1.8μm)column was used for gradient elution with 0.1%formic acid water and methanol(containing 0.1%formic acid)as mobile phase.The flow rate was 0.4 ml/min.The column temperature was 45℃.Positive ion scanning mode mass spectrometry with ESI source was used for detection.Results The linear range of the method was from 1 ng/ml to 2000 ng/ml,with regression coefficient of 0.9998.The limit of detection was 0.5 ng/ml.Recovery ranged from 93.3%to 96.4%,with RSDs of 0.86%-3.89%.Conclusion The method is simple,efficient,high sensitive and chemicals-saving and of high throughput,which can be applied to the determination of cotinine in the urine samples of smokers and second-hand smokers.