液相色谱-串联质谱法测定生物检材中乌头生物碱___

目的：建立检测生物检材中乌头生物碱的液相色谱-串联质谱(LC-MS/MS)法。方法：取血液、尿液或肝组织05 mL(g)，以丁丙诺啡为内标，经pH 92的硼砂溶液碱化后，用乙醚进行提取；采用Capcell Pak MG-Ⅱ C18柱(250 mm×20 mm，5 μm)，以乙腈-pH 4的20 mmol·L-1乙酸铵和01%甲酸的缓冲液(70∶30)为流动相，流速02 mL·min-1，采用电喷雾正离子模式离子化、多反应监测模式检测乌头生物碱，内标法定量。结果：血液、尿液和肝中乌头碱、次乌头碱、新乌头碱与内标丁丙诺啡色谱分离良好，乌头生物碱在01~50 ng·mL-1(ng·g-1)内均具有良好的线性，相关系数>0993，最低检出限为001~01 ng·mL-1(ng·g-1)，日内与日间精密度RSD均小于133%，血液中乌头生物碱提取回收率不小于648%。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、尿液及组织等生物检材中痕量乌头生物碱成分的检测。

Objective:To establish a method for determination of aconitum alkaloids in biological samples by LC-MS/MS.Methods:After 05 mL(g) of biological sample was extracted using a liquid-liquid extraction with diethyl ether at pH 92 and buprenorphine as an internal standard,aconitum alkaloids were separated on a Capcell Pak MG-Ⅱ C18 column (250 mm×20 mm，5 μm) using acetonitrile-buffer solution(20 mmol·L-1 ammonium acetate and 01% formic acid,pH 4) (70∶30) as the mobile phase at a flow-rate of 02 mL·min-1 with LC-MS/MS in the positive ionization mode.Quantitative results were obtained by the internal calibration procedure.Results:Aconitine,hypaconitine,mesaconitine and buprenorphine in blood,urine and liver were separated well.Calibration curve was linear within the range of 01-50 ng·mL-1(ng·g-1) with the LOD range from 001 to 01 ng·mL-1(ng·g-1).The inter-day and intra-day RSDs were less than 133%.The extraction recoveries of aconitum alkaloids in blood were more than 648%.Conclusion:This method is sensitive,fast and accurate for determination of trace aconitum alkaloids in biological samples.