超高压液相色谱-串联质谱法快速测定人尿液和全血中3种乌头生物碱

摘要

Objective To establish a rapid ultra-pressure liquid chromatography-tandem mass spectrome-try (UPLC-MS/MS) method for detection of aconitine, mesaconitine and hypaconitine in human urine and whole blood. Methods Urine sample was directly injected into the separation system and plasma sample was initially prepared by precipitation of proteins with acetonitrile–methanol (9:1, v/v). The analysis of the alkaloids was performed on an ACQUITY UPLC BEH C18 column using gradient elution with methanol and 5.0 mmol/L ammonium bicarbonate in water, and then detected by the positive electrospray ionization-MS/MS (ESI-MS/MS) method under MRM mode, and quantified by matrix internal standard method using noscapine as internal standard. Results The average recoveries were 93.9%~108%and 89.7%~109%for the three alkaloids in urine and whole blood, with relative standard deviation (RSD) of 1.3%~11%and 1.1%~18%(n=6), respec-tively. The quantitative limits (S/N=10) of the alkaloids in both of urine and whole blood were 0.05μg/L and 0.1μg/L. Conclusion This method is sensitive and accurate, and has been successfully applied to determine the 3 alkaloids in one case of food poisoning.%　　目的建立快速检测人尿液和全血中乌头碱、新乌头碱和次乌头碱的超高压液相色谱-串联质谱分析方法。方法尿液样品直接进样,全血样品经乙腈-甲醇(9：1, v/v)沉淀蛋白,以甲醇和5.0 mmol/L碳酸氢铵水溶液作为流动相进行梯度洗脱,在BEH C18色谱柱上实现分离,电喷雾正离子多反应监测方式检测,以那可丁作为内标的基质标准内标法定量。结果尿液和全血中3种乌头生物碱的平均加标回收率分别为93.9%~108%和89.7%~109%,相对标准偏差为1.3%~11%和1.1%∼18%(n=6),定量限(S/N =10)分别为0.05μg/L 和0.1μg/L。结论此方法灵敏、准确,已成功应用于一起中毒事件的检测。