高效液相色谱-质谱联用法同时测定全血中雪上一枝蒿中6种生物碱

赵万江; 杨福成; 裴雯; 冷晓晓; 施丝; 张茂林

doi:10.16369/j.oher.issn.1007-1326.2021.04.021

高效液相色谱-质谱联用法同时测定全血中雪上一枝蒿中6种生物碱

Simultaneous quantification of six alkaloids in aconitum brachypodum in whole blood by HPLC-MS/MS

摘要

摘要:
目的采用高效液相色谱-质谱联用法（HPLC-MS/MS）建立一种全血中同时测定雪上一枝蒿中6种生物碱（乌头碱、次乌头碱、新乌头碱、滇乌头碱、雪上一枝蒿甲素及雪上一枝蒿乙素）的快速、灵敏的检测方法。
方法采用Waters C18色谱柱，以甲醇-甲酸（两者体积分数为0.1%）水溶液为流动相，梯度洗脱，流速0.2 mL/min，乙腈蛋白沉淀法进行样品前处理，采用正离子-多反应离子监测模式对6种化合物进行测定。
结果乌头碱、次乌头碱、新乌头碱、滇乌头碱、雪上一枝蒿甲素及雪上一枝蒿乙素在0.5~100 ng/mL范围内线性关系良好，最低定量限依次为0.50、0.26、0.56、0.03、0.30、0.20 ng/mL，3个浓度的平均加标回收率为98.9%~105.8%，平均提取回收率为82.1%~90.3%，日内和日间精密度均 < 7%。
结论此方法基质效应良好，准确度高，灵敏度高，前处理简单，可同时、方便、快速检测雪上一枝蒿中6种生物碱，完成低含量的人体中毒毒物检测。

Abstract:
Objective To develop a HPLC-MS/MS method for simultaneous quantification of six alkaloids, namely aconitine, hypaconitine, mesaconitine, yunaconitine, bullatine A and bullatine B, in aconitum brachypodum in blood.
Methods The gradients were eluted with methanol-water with 0.1% formic acid as mobile phase and separated on Waters C18 column. The flow rate was 0.2 mL/min. Protein precipitation with acetonitrile was used in sample preparation. Electrospray ionization, multiple reaction monitoring scan mode were used for mass spectrometry detection.
Results The linear ranges were 0.5-100 ng/mL for six alkaloids. The limits of aconitine, hypaconitine, mesaconitine, yunaconitine, bullatine A and bullatine B were found to be 0.50, 0.26, 0.56, 0.03, 0.30 and 0.20 ng/mL, respectively. At three spiked concentrations, the average extraction recoveries ranged from 82.1%-90.3%, accuracy ranged from 98.9%-105.8%. The intra-day and inter-day RSDs were less than 7%.
Conclusions The method provides a sensitive, accurate, precise and reliable analytical procedure for clinical and toxicological monitoring of six alkaloids in aconitum brachypodum in whole blood.

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