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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。
OBJECTIVE: To optimize the extraction process of aristolochic acid Ⅰ in Radix et Rhizoma coptidis and explore the possibility of fungal fermentation to reduce its toxicity. Methods: L9 (34) orthogonal design was used to extract four factors including time, solvent fold, methanol concentration and extraction times, and then directly extracted by heating and refluxing method. HPLC and HPLC The contents of aristolochic acid Ⅰ in the fermented product solution of two kinds of medicinal fungi (F1 and F2) were as follows: acetonitrile-1% glacial acetic acid solution (51:49), the flow rate of 1.0 mL / min and the detection wavelength of 315 nm The column temperature was 30 ℃. Results: The optimum preparation process was powder 40 mesh, plus 10 times the amount of 60% methanol directly heated reflux extraction time 1h; compared with broad-spectrum herbs, the two fermentation products GF1 and GF2 aristolochic acid Ⅰ content were significantly decreased , The descending rate was 68.67% and 52.67% respectively, and new peaks appeared. Conclusion: The process of aristolochic acid Ⅰ extraction rate, good stability, suggesting that fungal fermentation may be broad-spectrum antifungus has some significance.