液相色谱-串联质谱法测定血液和尿液中秋水仙碱

目的建立检测血液和尿液中秋水仙碱的液相色谱-串联质谱法。方法0.5mL血液或尿液以丁丙诺啡为内标,经pH9.2硼酸盐缓冲溶液碱化后,用乙酸乙酯进行提取,在ZORBAX SB-C18液相柱(150mm×2.1mm×5μm)上以V(甲醇)∶V(20mmol/L乙酸铵和0.1%甲酸缓冲溶液)=80∶20为流动相,流速为0.2mL/min,采用电喷雾正离子模式离子化、多反应监测模式检测秋水仙碱,内标法定量。结果血液、尿液中秋水仙碱与内标丁丙诺啡色谱分离良好,秋水仙碱在0.1~50 ng/mL内均具有良好的线性,相关系数>0.9990,最低检出限为0.05ng/mL,方法回收率为94%~116%,日内与日间精密度(RSD)均小于8.5%。结论所建LC-MS-MS方法灵敏度高、操作简便、快速、准确,适用于血液及尿液等生物检材中痕量秋水仙碱成分的检测。

Determination of colchicine in urine and blood by liquid chromatography-tandem mass spectrometry with multiple-reaction monitoring

Objective To establish a method of liquid chromatography-tandem mass spectrometry for determination of colchicine in blood and urine.Methods 0.5mL of blood or urine sample was extracted using a liquid-liquid extraction(ethyl acetate) at pH 9.2 with buprenorphine as internal standard.The extract was separated on a ZORBAX SB-C18 column(150mm×2.1mm×5μm) using 20 mmol/L ammonium acetate with 0.1% formic acid(pH 4)/ methanol(20/80) as mobile phase with a flow-rate of 0.2mL/min,confirmed and quantified by MS-MS in the multiple reaction monitoring(MRM) mode via positive electrospray ionization mode(+ESI).Results Colchicine and buprenorphine in blood and urine were separated well.Calibration curves were linear within the range of 0.1～50ng/mL(r>0.9990),and the limits of detection were 0.05ng/mL.The recoveries were between 94%～116%,the inter-day and intra-day precisions were less than 8.5%.Conclusion This method is sensitive,fast and accurate,and suitable for determination of trace colchicine in blood and urine.