UPLC-QQQ-MS/MS法用于粉防己根双苄基异喹啉生物碱合成途径中12种化合物的定量检测

超高效液相色谱-三重四极杆串联质谱(UPLC-QQQ-MS/MS)法用于粉防己(Stephania tetrandra S.Moore)中的有机酸和生物碱类化合物的同时定性、定量检测具有重要的研究意义，可为汉防己甲素等双苄基异喹啉类生物碱的代谢合成机理解析提供数据参考。本研究采用Poroshell 120 CS-C18色谱柱进行分离，以含0.1%甲酸的10 mmol/L甲酸铵水溶液(A)-乙腈(B)为流动相进行梯度洗脱，在喷射流技术离子源(AJS-ESI)，正、负离子多反应监测(MRM)模式下进行检测，通过一级和二级质谱数据对不同采集批次的粉防己根部代谢物进行定性和定量分析。结果显示：12种被测化合物在线性范围内都呈现出良好的线性关系，相关系数均大于0.99；粉防己根部中12种被测化合物的含量存在显著差异，同一化合物的含量在不同的采集月份也有着明显的差异；L-酪氨酸、汉防己甲素和汉防己乙素在粉防己根部的含量相对较高。本研究实现了粉防己根部中12种被测化合物的同时定性和精确定量分析。

Quantitative Determination of 12 Compounds in the Biosynthetic Pathway of Bibenzylisoquinoline Alkaloids in the Roots of Stephania tetrandra S.Moore by UPLC-QQQ-MS/MS

It is of great significance for the simultaneous qualitative and quantitative detection of organic acids and alkaloids in Stephania tetrandra S.Moore by Ultra-High Performance Liquid Chromatography-Tandem Triple Quadrupole Mass Spectrometry (UPLC-QQQ-MS/MS) method.It can provide data reference for the analysis of the metabolic synthesis mechanism of bisbenzylisoquinoline alkaloids such as tetrandrine''s biosynthetic pathway in S. tetrandra S.Moore.In this study,a Poroshell 120 CS-C18 column was used for separation,with 10 mmol/L ammonium formate aqueous solution (A)-acetonitrile (B) containing 0.1% formic acid as the mobile phase for gradient elution,and the detection was carried out under the positive and negative ion Multiple Reaction Monitoring (MRM) mode of Agilent Jet Stream Electrospray Ionization (AJS-ESI).Qualitative and quantitative analysis of Stephania tetrandra S.Moore root metabolites in different batches was carried out by primary and secondary mass spectrometry data.The results showed that the 12 compounds showed a good linear relationship in the linear range,and the correlation coefficients were all greater than 0.99.In the roots of S. tetrandra S.Moore,there were significant differences in the content of 12 tested compounds,and the same compound also had significant differences in different collection months.The contents of L-tyrosine,tetrandrine and fangchinoline in the roots of S. tetrandra S.Moore were relatively high.In this study,the simultaneous qualitative and precise quantitative analysis of 12 compounds in S. tetrandra S.Moore have been realized.