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申润艳,陈衍炽,李检秀,蒙丽钧,严兴,李坚斌,谢能中.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[J].广西科学院学报,2023,39(4):461-470. [点击复制]
- SHEN Runyan,CHEN Yanchi,LI Jianxiu,MENG Lijun,YAN Xing,LI Jianbin,XIE Nengzhong.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[J].Journal of Guangxi Academy of Sciences,2023,39(4):461-470. [点击复制]
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| 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 |
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申润艳1, 陈衍炽2, 李检秀2, 蒙丽钧2, 严兴3, 李坚斌1, 谢能中2
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| (1.广西大学轻工与食品工程学院, 广西南宁 530003;2.广西科学院大健康所, 非粮生物质酶解国家重点实验室, 国家非粮生物质能源工程技术研究中心, 广西生物质工程技术研究中心, 广西生物炼制重点实验室, 广西南宁 530007;3.中国科学院分子植物科学卓越创新中心, 中国科学院合成生物学重点实验室, 上海 200032) |
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| 摘要: |
| 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。 |
| 关键词: 粉防己 超高效液相色谱-三重四极杆串联质谱 双苄基异喹啉生物碱 定性分析 定量分析 |
| DOI：10.13657/j.cnki.gxkxyxb.20231226.013 |
| 投稿时间：2023-09-19修订日期：2023-10-12 |
| 基金项目:国家自然科学基金项目(32260245)，广西科技重大专项(桂科AA2211700)，广西重大科技创新基地建设项目(2022-36-Z06-03，2022-36-Z06-07)，广西科技基地和人才专项(2021AC19176)，广西科学院创新团队启动基金(CQ-D-24215)和广西科学院发展基金项目(2021YFJ1210)资助。 |
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| 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 |
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SHEN Runyan1, CHEN Yanchi2, LI Jianxiu2, MENG Lijun2, YAN Xing3, LI Jianbin1, XIE Nengzhong2
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| (1.College of Light Industry and Food Engineering, Guangxi University, Nanning, Guangxi, 530003, China;2.State Key Laboratory of Non-food Biomass and Enzyme Technology, National Engineering Research Center for Non-food Biorefinery, Guangxi Biomass Engineering Technology Research Center, Guangxi Key Laboratory of Bio-refinery, Institute of Grand Health, Guangxi Academy of Sciences, Nanning, Guangxi, 530007, China;3.CAS-Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Shanghai, 200032, China) |
| Abstract: |
| 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. |
| Key words: Stephania tetrandra S. Moore ultra-high performance liquid chromatography-tandem triple quadrupole mass spectrometry bibenzylisoquinoline alkaloids qualitative analysis quantitative analysis |
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