中药黄芪中异黄酮苷类化合物的高效液相色谱-串联质谱分析

Analysis of Isoflavonoid Glycosides from A.mongholicus by Liquid Chromatography—Atmospheric Pressure Chemical Ionization Tandem Mass Spectrometry

利用高效液相色谱-大气压化学电离质谱联用技术,建立了黄芪提取物中的有效成分毛蕊异黄酮-7-O-β-D-葡萄糖苷和芒柄花素-7-O-β-D-葡萄糖苷的定性分析方法。这两个化合物在大气压化学电离质谱条件下,质谱裂解行为的研究表明,二者具有相似的特征质谱裂解行为,即在正离子模式下的一级质谱中均产生强的准分子离子[M+H]+峰和苷元离子[M+H-G lu]+峰;且苷元离子经碰撞诱导解离均产生.CH3(15 Da)、CH3OH(32Da)和2CO(56 Da)的中性丢失;同时发生Retro-D iels A lder(RDA)裂解反应,导致在毛蕊异黄酮-7-O-β-D-葡萄糖苷和芒柄花素-7-O-β-D-葡萄糖苷的二级质谱图中分别产生相对分子质量为m/Z148和m/Z133的特征碎片离子峰。这些特征碎片可以作为这两个化合物定性的依据。该方法已成功应用于黄芪注射液中的这两种物质的定性分析。

A method was established to characterize calycosin-7-O-β-D-glycoside and formononetin-7-O-β-D-glycoside, which had been proved to be associated with activity of Astragalus mongholicus （Huangqi）, by technique of liquid chromatography-atmospheric pressure chemical ionization tandem mass spectrometry （HPLC-APCI-MS/MS）. The study of fragment pressure chemical ionization indicates that both compounds have pathway pattern under atmospheric similar characteristic fragmentation behaviors as follows. The abundant quasi-molecular ions [ M ＋ H ] ^＋ and aglycone ions [ M ＋ H - Glu ] ^＋ can be observed in the positive MS spectrum. The collision-induced dissociation of aglycones produces the neutral loss of 15Da,32 Da and 56 Da corresponding to · CH3, CH3OH and 2CO,respectively. Simultaneously, the reaction of Retro-Diels Alder（RDA） cleavage takes place,in the course of which the characteristic fragment ions at m/Z 148 and m/Z 133 are generated corresponding to calycosin-7-O-β-D- glycoside and formononetin-7-O-β-D-glycoside, respectively. The characteristic fragment ions described above can be regarded as the important information for qualification of these two compounds. This method has been successfully applied to characterize these two compounds in the Huangqi injection.