正交试验优化白花蛇舌草4种五环三萜类成分提取工艺

Optimization of Extraction Technology of Four Pentacyclic Triterpenes from Hedyotis Diffusa by Orthogonal Experiment

目的优选白花蛇舌草五环三萜类成分山楂酸(MA)、科罗索酸(CA)、齐墩果酸(OA)、熊果酸(UA)的最佳提取工艺。方法采用HPLC建立MA、CA、OA、UA含量测定方法,以MA、CA、OA、UA含量为评价指标,通过单因素试验和正交试验,考察乙醇体积分数、提取时间、提取次数对提取效果的影响。结果 MA、CA、OA、UA色谱峰分离度良好,分别在0.568~3.408μg(R^(2)=0.999 0)、0.582~3.492μg(R^(2)=0.999 0)、0.488~2.928μg(R^(2)=0.999 9)、0.720~4.320μg(R^(2)=0.999 2)范围内呈良好的线性关系,平均加样回收率分别为99.09%(RSD=1.57%)、99.76%(RSD=1.78%)、100.49%(RSD=1.66%)、99.52%(RSD=1.75%)。最佳提取工艺条件为:采用95%乙醇提取70min,提取3次。结论本研究优选的提取工艺合理、稳定可行,可用于白花蛇舌草中MA、CA、OA、UA的提取。

Objective To optimize the extraction technology of pentacyclic triterpenes maslinic acid(MA), corosoic acid(CA), oleanolic acid(OA) and ursolic acid(UA) from Hedyotis diffusa. Methods A method for the determination of MA, CA, OA and UA was established by HPLC. The contents of MA, CA, OA and UA were set as evaluation indexes, and the effects of ethanol volume fraction, extraction time and extraction times on the extraction efficiency were investigated by single factor and orthogonal experiment. Results MA, CA, OA, UA had good resolution of chromatographic peak, showing good linear relationships in the ranges of 0.568–3.408 μg(R^(2)=0.999 0), 0.582–3.492 μg(R^(2)=0.999 0), 0.488–2.928 μg(R^(2)=0.999 9), 0.720–4.320 μg(R^(2)=0.999 2), respectively. The recovery rates were 99.09%(RSD=1.57%), 99.76%(RSD=1.78%), 100.49%(RSD=1.66%) and 99.52%(RSD=1.75%), respectively. The optimal extraction technology conditions were: 95% ethanol extraction for 70 min, 3 times for extraction. Conclusion The optimal extraction technology is reasonable, stable and feasible., and can be used to extract MA, CA, OA and UA from Hedyotis diffusa.