血浆中可替宁的固体介质液液萃取-液相色谱-串联质谱测定法

Determination of cotinine in plasma by supported liquid extraction-liquid chromatography-tandem mass spectrometry

目的建立一种快速准确测定血浆中可替宁的方法。方法将血浆样品碱化并加入内标后,通过96孔固体介质液液萃取对其进行分离纯化。经洗脱、氮气吹干、复溶后,使用Hypersil Gold C18 Selectivity色谱柱(50 mm×3 mm,1.9μm),以醋酸铵缓冲溶液-甲醇为流动相进行梯度洗脱,使用ESI源正离子扫描方式测定。结果本方法在0.50~200 ng/ml范围内线性良好(R2=0.999 5),定量检出限为0.25 ng/ml,加标回收率为84.0%~96.1%,RSDs为3.08%~7.66%(n=6)。结论该方法简单快捷,溶剂使用量小,具有高通量、高灵敏度的特点,适用于非吸烟人群及主动吸烟人群血浆中可替宁含量的测定。

Objective To establish a rapid method for determination of eotinine in plasma. Methods Plasma samples were alkalized and spiked with internal standard solution. After purification by SLE column, the samples were eluted, evaporated and reconstituted before tests. HypersilGo]d C18 Selectivity column （50 mm×3 mm, 1.9μm） was used for analysis. Gradient elution was performed using ammonium acetate buffer and methanol. Tandem mass spectrometric detection was employed at positive electrospray ionization. Results The eotinine solutions showed linear relationship in the range of 0.50-200 ng/ml （R2=0.9995）. The limit of quantification was 0.25 ng/ml. Recovery rates were 84.0%-96.1% with relative standard deviation of 3.08%-7.66% （n=6）. Conclusion This method has the advantages of easy and sample preparation, small amount of solvent used, and high sensitivity. It is suitable for the determination of cotinine levels in both smokers and non-smokers.