Box-Behnken响应面法优化苗药红花龙胆总黄酮的提取工艺及其含量测定研究

目的探讨红花龙胆中总黄酮的提取工艺及其含量测定的方法,并采用比色法测定贵州不同产地红花龙胆药材中总黄酮的含量差异。方法通过单因素试验研究甲醇浓度、溶剂用量、回流时间对红花龙胆中总黄酮提取率影响;以此为基础,采用Box-Bohnken Design试验设计原理进行三因素三水平试验设计和响应面分析法优化提取工艺;采用比色法,于410 nm处测定药材中以芒果苷计总黄酮的含有量。结果红花龙胆药材中总黄酮的最佳提取工艺为甲醇浓度78%、溶剂用量20.2 mL、回流时间140 min,应用优化后的提取工艺总黄酮得率为10.48%,与理论值10.52%的相对标准偏差为0.74%;不同产地红花龙胆药材中总黄酮的含量差异显著,含量范围为9.57%~20.91%。结论Box-Behnken响应面优化红花龙胆中总黄酮的提取工艺稳定、准确可靠,建立的总黄酮含量测定方法操作简便、重复性好、结果准确,可为红花龙胆药材的质量评价及相关研究提供参考。

Optimization of Extraction Process and Content Determination of Total Flavonoids from Guizhou Miao Medicine Gentiana Rhodantha Herba by Box-Behnken Response Surface Method

Objective To explore the methods of extraction process and the determination of the total flavonoids in Guizhou Miao Medicine Gentiana rhodantha herba and detect the contents of total flavonoids of Guizhou Miao Medicine Gentiana rhodantha herba from different producing areas in Guizhou by colorimetrie method.Methods The effects of methanol concentration,solvent dosage and reflux time on the extraction rate of total flavonoids from Guizhou Miao Medicine Gentiana rhodantha herba were studied by single factor experiments.On this basis,the three factors-three levels experiment was designed by Box-Bohnken Design centre combination theory.The optimal extraction conditions were obtained by using response surface methodology.The detection wavelength was 410 nm.The content of the total flavonoids in mangiferin was determined by colorimetry method.Results The optimal extraction process of total flavonoids from Guizhou Miao Medicine Gentiana rhodantha herba was as follows:the methanol concentration was 78%;the solvent dosage was 20.2 mL;the reflux time was 140 min.The total flavonoid yield of the optimized extraction conditions was 10.48%and the relative standard deviation between the predicted(10.52%)and measured value was 0.74%.The total content of total flavonoids from the different places was significantly different,ranging from 9.57%to 20.91%.Conclusion It was a stable,accurate and reliable method to obtain the total flavonoids from Guizhou Miao Medicine Gentiana rhodantha herba using the reflux optimization extraction technology process combined with Box-Benhnken design and response surface analysis.This method was simple,reliable and accurate,which could provide reference for the quality evaluation and related study of Guizhou Miao Medicine Gentiana rhodantha herba.