Box-Behnken响应面法优化虎杖提取工艺及其质量标准研究

Extraction Technology Optimization for Polygonum Cuspidatum by Box-Behnken Response Surface Method and Its Quality Standard Research

目的:优化虎杖提取工艺,在此基础上建立虎杖提取物的质量标准。方法:以虎杖苷、白藜芦醇、大黄素含量的综合评分(Y)为评价指标,采用Box-Behnken响应面法对虎杖提取工艺的乙醇浓度、提取时间、提取温度进行筛选。根据优化工艺制备10批来自不同产地的虎杖提取物,采用TLC建立提取物中虎杖苷、大黄素的定性鉴别方法,并计算10批提取物的含水量。运用HPLC得到10批提取物中虎杖苷、白藜芦醇、大黄素的含量,并建立提取物指纹图谱。结果:响应面法优化出的工艺为乙醇浓度60%、提取时间75 min、提取温度70℃。TLC结果显示提取物中虎杖苷、大黄素的斑点清晰,分离度好。10批提取物的水分含量范围为2.83%-5.17%。虎杖苷、白藜芦醇、大黄素的含量范围分别为15.19-36.36 mg/g、0.95-5.71 mg/g、5.20-15.47 mg/g。10批虎杖提取物的指纹图谱中出现了11个共有峰,与对照指纹图谱的相似度均高于0.90。结论:优化出的虎杖提取工艺稳定可行,在此基础上建立的虎杖提取物质量标准可重复性和稳定性良好。初步拟定虎杖提取物中水分不得过6.00%。按干燥品计算,虎杖苷、白藜芦醇、大黄素含量分别不得少于15.10、0.90、5.00 mg/g。

Objective:To optimize the extraction process of Polygonum cuspidatum and establish the quality standard of extract.Methods:The comprehensive score(Y)of the content of polydatin,resveratrol and emodin was taken as the evaluation index,and Box Behnken response surface method was used to screen the ethanol concentration,extraction time and extraction temperature of the extraction process of Polygonum cuspidatum.According to the optimized process,10 batches of extracts from different origins were prepared.Qualitative identification methods of polydatin and emodin were established by TLC,and the moisture content of 10 batches of extracts was calculated.The contents of polydatin,resveratrol and emodin in 10 batches of extracts were determined by HPLC,and the fingerprint of the extracts was established.Results:The optimal extraction process was that the ethanol concentration was 60%,the extraction time was 75 min,and the extraction temperature was 70℃.The TLC results showed that the spots of polydatin and emodin in the extract were clear and well separated.The moisture content of 10 batches of extracts ranged from 2.83%to 5.17%.The content of polydatin ranged from 15.19 mg/g to 36.36 mg/g,the content of resveratrol ranged from 0.95 mg/g to 5.71 mg/g,the content of emodin ranged from 5.20 mg/g to 15.47 mg/g,respectively.There are 11 common peaks in the fingerprints of 10 batches of extracts,and the similarities with the control fingerprint were all higher than 0.90.Conclusion:The optimized extraction process of Polygonum cuspidatum is stable and feasible,and the quality standard of Polygonum cuspidatum extract has good repeatability and stability.It is preliminarily proposed that the moisture content of Polygonum cuspidatum extract should not exceed 6.00%.The contents of polydatin,resveratrol and emodin shall not be less than 15.10 mg/g,0.90 mg/g and 5.00 mg/g respectively,according to the dry product.