利用质谱研究诃子对草乌的化学解毒作用

Chemical Detoxification of Fruit of Medicine Terminalia on Root of Kusnezoffii Monkshood by Mass Spectrometry

摘要:

目的: 揭示蒙古族医药理论中诃子对草乌解毒作用的科学内涵. 方法: 采用超声提取法对草乌、草乌-诃子药对进行乙醚、无水乙醇提取,对草乌进行酸性乙醚提取,对草乌中双酯型生物碱提取部位-诃子混合后进行乙醚提取;采用电喷雾质谱技术对上述提取液进行检测. 结果: 电喷雾一级质谱图结果显示,随着诃子量的增加,草乌-诃子乙醚提取液中双酯型生物碱的相对丰度呈下降趋势,而脂型生物碱相对丰度呈上升趋势;草乌酸性乙醚提取液中脂型生物碱为基峰;草乌与草乌-诃子无水乙醇提取液中同种生物碱的相对丰度基本一致;双酯型生物碱提取部位加入诃子后,各生物碱的相对丰度与种类均无变化. 结论: 蒙古族医药理论中诃子对草乌的解毒作用是指在低极性条件下,诃子通过抑制双酯型生物碱的溶出对草乌起到减毒作用.本实验方法可用于制剂工艺为药材粉末直接混合制成的各种蒙古族成药复方的配伍机制研究.

Abstract:

Objective: The aim of this investigation was to study the detoxification of fruit of Medicine Terminalia (FMT) on root of Kusnezoffii Monkshood (RKM). Method: Ultrasonic extraction was applied to extract aconite alkaloids in RKM and RKM-FMT powder. ESI-MS was adopted to monitor the alkaloids in diethyl ether extract of RKM and the blend of RKM with FMT, in acidic diethyl ether extract of RKM, in absolute ethanol extract of RKM and the blend of RKM with FMT, in absolute ethanol extract of the residue of blend of RKM with FMT after extracting by diethyl ether, as well as diethyl ether extract of the blend of diester-alkaloids with FMT. Result: There were monoester-alkaloids, diester-alkaloids and lipo-alkaloids in diethyl ether extract of RKM, and the relative abundance of diester-alkaloids was the highest among them. In diethyl ether extract of the blend of RKM with FMT, the relative abundance of diester-alkaloids decreased as the FMT amount increased, moreover, that of lipo-alkaloids became the highest when the FMT amount was three times as that of RKM. In acidic diethyl ether extract of RKM, the lipo-alkaloids were detected as the basic peak. The relative abundance of all alkaloids in absolute ethanol extract of RKM was the same as that of the blend of RKM with FMT. FMT didn't affect the relative abundance of the alkaloids in diethyl ether extract of the blend of diester-aconite alkaloids with FMT. On the basis of the above results, we could conclude that FMT can restrain the dissolution of dister-alkaloids of RKM in diethyl ether, however, in absolute ethanol the effect was related to the amount of added FMT. Acid condition could also restrain the dissolution of dister-alkaloids in diethyl ether. Conclusion: Under low polarity condition, detoxification of FMT on RKM was through restraining the release of diester-alkaloids in diethyl ether. This research is significant for studying antidotal method of RKM as well as clarifying the mechanism of synergy of formula which contains both RKM and FMT.