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目的探讨黄连解毒汤(HJD)自沉淀制备条件,测定自沉淀主要成分量及自沉淀对PC12细胞损伤模型的保护作用。方法以提取时间和投水量为影响因素,确定自沉淀的制备工艺;采用HPLC法,测定自沉淀中黄芩苷、小檗碱、栀子苷、巴马汀、黄连碱、汉黄芩苷的量;采用CCK-8法评价HJD上清液、自沉淀对CoCl_2损伤的PC12细胞模型作用。结果在10倍量水回流提取30 min的煎煮条件下,干燥后HJD自沉淀质量约占上清液质量的10%,自沉淀中81.28%的成分为酸碱性化合物,其中黄芩苷占42.12%,小檗碱占31.17%,黄连碱占5.89%,汉黄芩苷占1.50%,巴马汀占0.60%,未检测到中性成分栀子苷。上清液、自沉淀对CoCl_2损伤的PC12细胞的EC50值分别28.25、19.58μg/m L。结论自沉淀制备工艺操作简单,重复性好,通过成分分析,得出自沉淀的形成机制以酸碱络合为主的假想;与上清液相比,自沉淀表现出较好的神经保护活性,为HJD的二次开发提供参考。
OBJECTIVE To investigate the preparation conditions of self-precipitation of Huangjian Jiedu Decoction (HJD), and to determine the main components of self-precipitation and the protective effect of self-precipitation on the injury model of PC12 cells. Methods The extraction process and the amount of water were taken as the influencing factors to determine the preparation process of self-precipitation. The contents of baicalin, berberine, geniposide, palmatine, coptisine and baicalin in the self-precipitation were determined by HPLC. CCK-8 method was used to evaluate the effect of HJD supernatant on the PC12 cell model induced by CoCl2. Results The quality of HJD self-precipitation was about 10% of the supernatant after drying under the condition of 10 times the amount of water reflux extraction for 30 min, and the content of 81.28% of the self-precipitation was acidic and basic compounds, of which baicalin accounted for 42.12 %, Berberine accounted for 31.17%, Coptis alkaloid accounted for 5.89%, Han baicalin accounted for 1.50%, palmatine accounted for 0.60%, did not detect neutral ingredients Geniposide. The EC50 values of supernatant and self-precipitated PC12 cells damaged by CoCl 2 were 28.25 and 19.58 μg / m L, respectively. CONCLUSIONS The self-precipitation process has simple operation and good repeatability. Through the analysis of composition, it is concluded that the mechanism of self-precipitation is acid-base complexation. Compared with the supernatant, the self-precipitation shows good neuroprotective activity, HJD secondary development for reference.