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

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-C_(18)(2.1 mm×50 mm,2.7μm),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。

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-C_（18）（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 of10-100 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%（RSD10%,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.