基于药效学跟踪试验提取分离龙须藤总黄酮及其含量测定

Extraction, Separation and Determination of Total Flavonoids from Bauhinia chaampioni Based on Pharmacodynamics Tracking Test

摘要:

目的: 考察龙须藤的最佳乙醇提取浓度,并以硅胶柱色谱法分离得到龙须藤总黄酮,对分离得到的龙须藤总黄酮进行含量测定。方法: 采用药效学跟踪试验,参考龙须藤不同浓度乙醇提取物对膝关节炎症模型大鼠的抗炎作用,选取龙须藤的最佳乙醇提取浓度,采用硅胶柱色谱方法从龙须藤乙醇提取物的乙酸乙酯萃取部位分离得到龙须藤总黄酮,HPLC对分离得到的龙须藤总黄酮进行含量测定。结果: 龙须藤的70%乙醇提取物抗炎作用最好,采用溶剂系统为二氯甲烷-乙酸乙酯(8:1~4:1)梯度洗脱分离得到的龙须藤总黄酮中3',4'-亚甲二氧基-5',5,6,7-四甲氧基黄酮、3',4',5',5,7-五甲氧基黄酮含量分别为21.3%,46.7%。结论: 龙须藤的最佳乙醇提取浓度为70%,溶剂系统为二氯甲烷/乙酸乙酯(8/1~4/1)梯度洗脱分离龙须藤总黄酮的方法可行。

Abstract:

Objective: To optimize the concentration of ethanol to extract total flavonoids from Bauhinia championii, and purified using silica gel column chromatography. Quantitative determine the content of two major flavonoids in total flavonoids. Method: The optimized concentration of ethanol was selected based on anti-inflammatory effects of extract from B. chaampioni on knee osteoarthritis model in rats. The total flavonoids were enriched by silica gel column chromatography. The contents of two flavonoids, 3',4'-methylenedioxy-5',5,6,7-tetramethoxyflavone and 3',4',5',5,7-pentamethoxyflavone was determined by HPLC. Result: The 70% ethanol extract of B. chaampionihad the strongest anti-inflammatory effect. The solvent system used in silica gel column chromatography was dichloromethane-ethyl acetate (8:1 to 4:1). The content of 3',4'-methylenedioxy-5',5,6,7-tetramethoxyflavone and 3',4',5',5,7-pentamethoxyflavone in total flavonoids were 21.3% and 46.7%, respectively. Conclusion: The established route to obtain total flavonoid from B. chaampioni was feasible.