超高效液相色谱-飞行时间质谱法定性定量检测乌头类植物中14种乌头生物碱及其 代谢产物的含量

Qualitative and quantitative determination of 14 kinds of aconitine alkaloids and their metabolites in Aconitum L. by ultra performance liquid chromatography-quadrupole-time of flight mass spectrometer

中文关键词:  超高效液相色谱-飞行时间质谱法  定性定量分析  乌头类植物  乌头碱

英文关键词:ultra performance liquid chromatography-quadrupole-time of flight mass spectrometer  qualitative and quantitative analysis  Aconitum L.  aconitine alkaloids

中文摘要:

      目的 建立超高效液相色谱-飞行时间质谱(ultra performance liquid chromatography- quadrupole-time of flight mass spectrometer, UPLC/Q-TOF)同时定性定量检测乌头类植物中14种乌头生物碱及其代谢产物的含量的分析方法。方法 采用SPE固相萃取小柱提取净化, 用超高效液相色谱串联四级杆飞行时间质谱, Dual AJS ESI源, 正离子模式下采集的数据进行定性定量分析。色谱柱为Aglient SB-C18(2.1 mm×50 mm, 2.7 ?m), 流动相为乙腈-0.1%甲酸水(含5 mmol/L乙酸铵), 梯度洗脱, 流速0.3 mL/min。结果 14种乌头生物碱及其代谢产物在10~100 ng/mL范围内具有良好的线性关系, 相关系数均大于0.99, 检出限为0.1~5.0 ng/mL, 定量限为0.5~10 ng/mL, 加标回收率为81.2%~113.7%(相对标准偏差(relative standard deviation, RSD)<10%, n=6)。 结论 该方法能快速、准确地对乌头类植物中的乌头生物碱成分进行定性定量检测, 对该植物毒性和药效的深入研究提供了一种新的方法。

英文摘要:

      Objective To establish a method for qualitative and quantitative analysis of 14 kinds of aconitine alkaloids and their metabolites in Aconitum L. by ultra performance liquid chromatography-quadrupole-time of flight mass spectrometer (UPLC/Q-TOF). Methods The samples were extracted and purified by SPE solid phase extraction cartridge, and the data was collected by ultra-high performance liquid chromatography tandem flight time mass spectrometry with Dual AJS ESI source, and analyzed qualitatively and quantitatively in positive ion mode. Chromatographic column was Agilent SB-C18 (2.1 mm×50 mm, 2.7 ?m) and the flow phase was acetonitrile-0.1% formic acid aqueous solution containing 5 mmol/L ammonium acetate (gradient elution) at a flow rate of 0.3 mL/min. Results The 14 kinds of aconitum alkaloids and their metabolites had good linear relationship in the range of 10-100 ng/mL, and the correlation coefficients were greater than 0.99. The limits of detection were 0.1-5.0 ng/mL, the limits of quantification were 0.5-10 ng/mL, and the recovery rates were 81.2%-113.7% (RSD<10%, n=6). Conclusion This method can quickly and accurately determine the qualitative and quantitative detection of aconite alkaloids in aconite plants, which provides a new method for the in-depth study of the phytotoxicity and efficacy of the plant.