基于抗乙酰胆碱酯酶活性及多成分含量分析的天泰1号提取工艺优化

Optimization of Extraction Technology on Tiantaiyihao by Multi-index Comprehensive Evaluation of Anti-AChE Activity and Bioactive Components’ Contents

目的优化主治老年痴呆症的天泰1号(黄连、人参、天麻等组成)的提取工艺。方法以乙酰胆碱酯酶抑制率、浸膏得率、黄连生物碱和人参皂苷含量加权综合评分为指标优选工艺;采用改进的Ellman法测定抑制乙酰胆碱酯酶活性;采用高效液相色谱法(HPLC)测定主药黄连生物碱(小檗碱、表小檗碱、黄连碱、巴马汀)和人参皂苷(人参皂苷Rg1、Re、Rf、Rb1、Rd)的含量;在单因素考察基础上,采用正交设计方法,考察乙醇浓度、溶媒用量、提取时间、浸泡时间对提取工艺的影响。结果改良Ellman法测定乙酰胆碱酯酶方法操作简单、结果准确可靠;黄连生物碱和人参皂苷检测方法线性关系良好(R2>0.999 5),平均加样回收率95.44%~101.50%,相对标准偏差(RSD)<2.96%;优选最佳提取工艺为采用70%乙醇为溶媒,加入10倍溶媒,提取2次,每次1.5 h,最优提取工艺参数组合验证得分0.91分,所得提取物具有良好的抑制乙酰胆碱酯酶活性作用。结论基于活性评价和药效成分多指标优化的提取工艺参数具有科学性、合理性,适合产业化生产。

Objective To optimize the extraction process of Tiantaiyihao. Methods The weighting scores with the acetylcholinesterase activity inhibition rate, extraction yield, contents of alkaloids from Coptidis rhizoma and ginsenoside were used as indexes to optimize the extraction process. The modified Ellman method was applied to determine the inhibition activity of Acetylcholinesterase; HPLC method was used to determine the contents of alkaloids(Berberine,Epiberberine,Coptisine,Palmatine)and ginsenoside(ginsenoside Rg1,Re,Rf,Rb1,Rd)in Tiantaiyihao. Orthogonal design method was used to optimize the volume and concentration of alcohol,extraction time and soaking time. Results The modified Ellman method was simple and reliable in determining inhibition activity of acetylcholinesterase. The linear relationships of content determination methods were good(R2>0.999 5)and the average recoveries were 95.44%-101.50% with RSD<2.96%. The optimum extraction parameters are:reflux extracting for 2 times with 10-folds 70% ethanol,1.5 hours each time. The validation score of optimal combination was 0.91. The inhibition of acetylcholinesterase activity for Tiantaiyihao by optimal extraction process was good.Conclusion The optimized extraction process parameters based on the activity evaluated and bioactive components’ contents evaluation,was scientific,reasonable and suitable for industrial production.