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

Determination of 3 kinds of Aconitum alkaloids in human urine and whole blood using ultra-pressure liquid chromatography-mass spectrometry

中文关键词:  超高压液相色谱-串联质谱法  乌头碱  新乌头碱  次乌头碱  尿液  全血

英文关键词:ultra-pressure liquid chromatography-tandem mass spectrometry (UPLC-MS/MS)  aconitine  mesaconitine  hypaconitine  urine  whole blood

中文摘要:

      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 0.05 μg/L 和 0.1 μg/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。

英文摘要:

      Objective To establish a rapid ultra-pressure liquid chromatography-tandem mass spectrometry (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), respectively. The quantitative limits (S/N=10) of the alkaloids in both of urine and whole blood were 0.05 μg/L and 0.1 μg/L. Conclusion This method is sensitive and accurate, and has been successfully applied to determine the 3 alkaloids in one case of food poisoning.