苦参不同组织中生物碱类成分的LC-MS分析

LC-MS analysis of quinolizidine alkaloids in different tissues of Sophora flavescens

目的 通过对苦参Sophora flavescens不同组织样品中生物碱类成分的LC-MS分析,探讨苦参中生物碱类成分的质谱裂解规律及其在不同样品中的异同。方法 首先利用UPLC-Q-Exactive-OrbitrapMS对5年生苦参根中的生物碱类成分进行全面的鉴定分析。再利用UPLC-QQQ-MS/MS技术,对苦参无菌幼苗不同组织部位(根、茎、叶)、苦参愈伤组织中的生物碱类成分进行定量分析,比较各样品中生物碱类成分的异同。结果 从5年生苦参根中共鉴定出47个生物碱类成分;利用UPLC-QQQ-MS/MS对其中17个主要的生物碱类成分进行了定量分析,发现不同苦参样品中生物碱类成分差异明显,其中苦参无菌幼苗各组织部位中生物碱的含量相对较高。结论 建立了一种苦参生物碱类成分快速定性定量分析方法,从5年生苦参根中快速鉴定出47种生物碱;通过对苦参不同组织样品中17个生物碱类成分的定量分析,为后续利用比较转录组测序技术,挖掘并鉴定苦参中生物碱类成分生物合成关键酶的研究奠定基础。

Objective To explore the mass spectrometric cleavage patterns of the alkaloids in Sophora flavescens and their variation in different S.flavescens samples,including aseptic seedlings(roots,leaves,and stems)and callus,by LC-MS analysis,so as to provide the basis for the identification of the key enzymes involved in the biosynthetic pathway of S.flavescens alkaloids using comparative transcriptome sequencing approach.Methods UPLC-Q-Exactive-Orbitrap MS was used to comprehensively identify the alkaloids in the roots of 5-year-old S.flavescens.UPLC-QQQ-MS/MS was used to quantify the alkaloids in different tissues(roots,stems,leaves)of S.flavescens aseptic seedling and callus.Results A total of 47 alkaloids components were identified from the roots of 5-year-old S.flavescens,and 17 of them were quantified by UPLC-QQQ-MS/MS.The results revealed that the alkaloids in different S.flavescens samples were significantly different,and the content of alkaloids in tissues of S.flavescens aseptic seedling was relatively high.Conclusion In this study,a rapid qualitative and quantitative method for the alkaloids in S.flavescens was developed,resulting in the identification of 47 alkaloids from S.flavescens roots,and 17 alkaloids in different S.flavescens tissue samples were quantified,which are helpful to the subsequent mining and identification of key enzymes involved in the biosynthesis of alkaloids in S.flavescens by using comparative transcriptome sequencing technology.