超高效液相色谱-串联质谱法快速检测自制草乌药酒中9种乌头类生物碱含量

Rapid determination of 9 kinds of aconitum alkaloids in homemade Radix aconiti Kusnezoffii medicinal liquor by ultra performance liquid chromatography-tandem mass spectrometry

中文关键词:  滇草乌  自制药酒  乌头类生物碱  快速筛查  超高效液相色谱串联质谱

英文关键词:Yunnan Radix aconiti Kusnezoffii  homemade medicinal liquor  aconitum alkaloids  rapid screening  ultra performance liquid chromatography-tandem mass spectrometry

中文摘要:

      目的 建立超高效液相色谱-串联质谱法快速测定自制草乌药酒中9种乌头类生物碱含量的方法。方法 以云南野生的滇草乌为主要原料, 将草乌制成5种不同形状(整块、去皮、片状、块状、粉末状), 分别取250 g浸泡于900 mL 52度玉米酿制白酒中, 浸泡时间约为1.5年。移取1 mL药酒样品, 加入30%甲醇水溶液(3:7, V:V)至100 mL, 经0.22 μm滤膜过滤后备测。利用C18色谱柱和乙腈-0.1%甲酸水的流动相进行梯度洗脱分离, 多反应监测模式检测, 通过基质工作曲线外标法定量。结果 9种乌头类生物碱在1~100 μg/L范围内线性关系良好, 相关系数均大于0.999, 检出限为0.63~0.98 μg/L, 定量限为2.11~3.26 μg/L, 低、中、高3种浓度水平下加标回收率为81.14%~116.01%, 相对标准偏差为0.97%~11.26% (n=6)。经实际样品测定分析, 药酒中9种乌头类生物碱的双酯型生物碱含量居高, 其中以乌头碱含量最多, 毒性较强。结论 该方法前处理简便, 灵敏度高, 准确性好, 检测效率高, 适用于卫生应急中毒事件中自制草乌药酒9种乌头类生物碱的快速筛查, 为医疗救治的快速诊断和采取对应急救治疗措施提供强有力技术支撑。

英文摘要:

      Objective To establish a method for the rapid determination of the content of 9 kinds of aconitum alkaloids in homemade Radix aconiti Kusnezoffii medicinal wine by ultra performance liquid chromatography-tandem mass spectrometry. Methods The wild Radix aconiti Kusnezoffii in Yunnan was used as the main raw material. It was made into 5 kinds of different shapes (whole, peeled, flake, block, powder), and 250 g was soaked in 900 mL of 52-degree corn to brew liquor, and the soaking time was about 1.5 years. Took 1 mL sample of medicinal liquor, added 30% methanol water solution (3:7, V:V) to 100 mL, and filtered it with 0.22 μm filter membrane, ready for testing. C18 column and mobile phase of acetonitrile-0.1% formic acid water were used for gradient elution separation, multi-reaction monitoring mode detection, and quantification was carried out by matrix working curve external standard method. Results The linear relationship of 9 kinds of aconitum alkaloids was good in the range of 1–100 μg/L, the correlation coefficients were all greater than 0.999, the limits of detection were 0.63–0.98 μg/L, the limit of quantitation were 2.11–3.26 μg/L, and the recovery rates were 81.14%–116.01% at low, medium and high concentrations, the relative standard deviations were 0.97%–11.26% (n=6). According to the analysis of actual samples, the content of diester alkaloids of 9 kinds of aconitum alkaloids in medicinal liquor was high, among which aconitine had the most content and strong toxicity. Conclusion This method is simple in pretreatment, high in sensitivity, good in accuracy and high in detection efficiency. It is suitable for rapid screening of 9 kinds of aconitum alkaloids in Radix aconiti Kusnezoffii homemade medicinal liquor in health emergency poisoning events, and provides strong technical support for rapid diagnosis of medical treatment and corresponding emergency treatment measures.