马钱子微量生物碱的ESI-MS/MS质谱分析

目的： 分析中药马钱子总生物碱提取物中微量生物碱16-羟基-α-可鲁勃林（16-OH-α-colubrine）、异番木鳖碱（isostrychnine）质谱碎裂规律及与结构间的关系。在此基础上建立区分马钱子中微量生物碱同分异构体番木鳖碱（strychnine）和异番木鳖碱、16-羟基-α-可鲁勃林和4-羟基依卡精（4-OH-vomicine）的简便、快速、灵敏的新方法。方法： 采用硅胶色谱层析柱（6 dm×120 dm），用正己烷-三氯甲烷-二乙胺（5：4：0.3）和（5：5：0.3）梯度洗脱，对马钱子总生物碱提取物进行分离，应用电喷雾质谱全扫描及串联质谱技术（ESI-MS/MS）对R₇洗脱流份进行分析，通过相对分子质量和多级串联质谱的信息对马钱子中微量生物碱的质谱碎裂规律进行分析研究。结果： 归属并确认了R₇洗脱流份中微量生物碱16-羟基-α-可鲁勃林、异番木鳖碱的存在，并初步提出2种微量生物碱的质谱碎裂机理；在其质谱碎裂规律的基础上建立了简便、快捷地区分微量马钱生物碱异构体16-羟基-α-可鲁勃林和4-羟基依卡精、番木鳖碱和异番木鳖碱的方法。结论： 本实验方法为马钱生物碱类化合物快速、灵敏分析方法的建立提供了重要依据。

Objective: To analyze the fragmentation rule of 16-OH-α-colubrine and isostrychnine in total alkaloid extracts of Strychni Semen by mass spectrometry and their relationship with structure. On this basis,a simple,rapid and sensitive method was established to distinguish the isomers of strychnine and isostrychnine,16-OH-α-colubrine and 4-OH-vomicine in Strychni Semen. Methods: The total alkaloid extracts from Strychni Semen were separated on a silica gel column (6 dm×120 dm)with gradient elution of n-hexane-chloroform-diethylamine at ratio of (5:4:0.3)and (5:5:0.3). The R₇ elution fraction was analyzed by electrospray ionization mass spectrometry (mass spectrometry)and tandem mass spectrometry (ESI-MS/MS). The fragmentation law of alkaloids by mass spectrometry was analyzed and studied. Results: The existence of trace alkaloids of 16-OH-α-colubrine and isostrychnine in the elution fraction of R₇ was confirmed. Based on the rule of mass spectrometry,a simple and rapid method was established to distinguish trace amount of strychnine alkaloid isomers,such as 16-OH-α-colubrine,4-OH-vomicine,strychnine and isostrychnine. Conclusion: This method provides an important basis for the establishment of a rapid and sensitive method for the analysis of strychnine alkaloids.

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