广防己中马兜铃酸Ⅰ的提取工艺优选及真菌发酵对其含量的影响

目的:优化广防己中马兜铃酸Ⅰ的提取工艺,并探讨真菌发酵降低毒性作用的可能性。方法:采用L9(34)正交设计,以提取时间、溶剂倍数、甲醇浓度、提取次数4个因素为考察指标,直接加热回流法提取,采用高效液相色谱法(HPLC)测定广防己及其两种药用真菌(F1菌和F2菌)发酵品溶液中马兜铃酸Ⅰ的含量,流动相为乙腈-1%冰醋酸水溶液(51:49),流速为1.0mL/min,检测波长315nm,柱温30℃。结果:最佳制备工艺为粉末40目,加10倍量60%甲醇直接加热回流提取一次1h;与广防己药材相比,两种发酵品GF1和GF2中马兜铃酸Ⅰ的含量均明显下降,下降率分别为68.67%和52.67%,且有新的色谱峰出现。结论:该工艺马兜铃酸Ⅰ提取率高,稳定性好,提示真菌发酵可能对广防己的减毒具有一定的意义。

Optimization of extraction technique of aristolochic acid I in Aristolochia fangchi and influence of fungal fermentation on its content

Objective: To optimize the extraction technique of aristolochic acid I in Aristolochia fangchi and explore the possibility of reducing toxicity by fungal fermentation.Methods: Orthogonal design L9(34) was adopted,the contents of aristolochic acid I in crude drug and its two fermented products which were boiled under a reflux condenser were determined with the extraction time,multiple of solvent,methanol concentration and times of extraction as the observation indexes.The mobile phase was a mixture of acetonitrile and 1% acetic acid aqueous solution(51:49).The flow rate was 1.0 mL/min.The detection wavelength was at 315nm and the column temperature was at 30℃.Results: The optimum conditions were that the forty mesh powder was refluxed only once for 1h with ten times of 60% methanol.Compared with the crude drug,the content of aristolochic acid I in two fermentation products(GF1 and GF2) decreased significantly,the rate of decline was 68.67% and 52.67% respectively.In addition,new peaks occurred.Conclusion: The technique screened has characteristics of high extraction efficiency and stability.The results indicates that it maybe meaningful to reduce its toxicity on Aristolochia fangchi by fungal fermentation method.