高乌头不同溶剂萃取部分HPLC多波长指纹图谱与其急性毒性的谱-毒研究

Spectrum and Toxicity Relationships between Multi-Wavelength HPLC Fingerprints and Acute Toxicity of Different Extracts of Aconitum sinomontanum Nakai

目的建立高乌头药材不同极性部分HPLC多波长指纹图谱(MWCF),阐明高乌头不同溶剂萃取部分的指纹图谱特征峰所代表的化学成分,及其对急性毒性贡献的大小。方法采用系统溶剂法对不同产区高乌头药材体积分数95%乙醇提取物进行萃取,依次得到石油醚、三氯甲烷、乙酸乙酯、正丁醇、乙醇和水提6个不同的溶剂萃取部分;运用HPLC采用均谱法(MFM)建立高乌头不同溶剂萃取部分的MWCF;用测定半数致死量(LD_(50))和最大给药量的方法对高乌头不同溶剂萃取部分的急性毒性进行比较研究;用灰关联分析法建立其谱-毒关系。结果建立了高乌头药材不同溶剂萃取部分的MWCF,高乌头的毒性成分不只是高乌甲素与冉乌头碱,其他二萜类生物碱贡献也不可忽视,各特征峰所代表的化学成分对其毒性贡献大小的顺序(按特征峰编号)为(51、38、37、35、20号)> 34号> 39号> 32号> 31号> 33号。结论采用MFM所构建的高乌头不同溶剂萃取部分MWCF能最大限度保留指纹峰,实现化学指纹信息最大化,同时能有效改善指纹信号质量,可为高乌头药材的整体质量评价提供参考。不同溶剂萃取部分MWCF与其急性毒性之间有一定的对应关系,可以为高乌头的毒性物质基础研究提供参考。

OBJECTIVE To establish the HPLC multiple wavelength chromatographic fingerprints (MWCF) of different extracts of Aconitum sinomontanum Nakai (ASN) to clarify the attribution of the fingerprint peaks and their contribution to the acute toxicity. METHODS The experimental drugs (extracts of petroleum ether, chloroform, ethyl acetate, butanol, alcohol and water) were obtained by means of systematic solvent extraction from the 95% ethanol extract of ASN. The HPLC MWCF of different extracts of ASN were established by mean fingerprint method (MFM). The acute toxicity of different extracts in mice were carried out by measuring the median lethal dose (LD50) and maximum dose. The relationship between spectrum and toxicity was established by gray relational analysis. RESULTS The MWCF of different extracts of ASN were established. The acute toxicity of ASN was caused not only by lappacontine (LAP) and ranaconitine, the contribution of other diterpenoid alkaloids should not be neglected, and the contribution of different peaks to toxicity was ranked as (51, 38, 37, 35, 20) >34>39>32>31>33. CONCLUSION The MWCF developed by MFM can maximally retain the fingerprint peaks, achieve fingerprint information maximization, and effectively improve fingerprint signal quality, thus providing a reference for the comprehensive quality evaluation of ASN. The relationship between the MWCF of different extracts and the acute toxicity is paralleled to some extent. And this will lay a foundation for the research of the toxicity mechanism of ASN.