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目的:探索D-101大孔树脂分离纯化岩柏草总黄酮的最优工艺,并对有效成分穗花杉双黄酮进行含量测定。方法:紫外分光光度法测定总黄酮浓度,以黄酮收率、纯度等为指标,上样质量浓度、吸附流速和洗脱剂乙醇体积分数为影响因素,确定大孔树脂分离纯化岩柏草总黄酮工艺,并用高效液相色谱法测定总黄酮中穗花杉双黄酮。结果:大孔树脂对岩柏草总黄酮有较好的分离效果,其最优工艺条件为上样质量浓度8.5mg/mL,吸附流速为2mL/min,乙醇体积分数为70%,总黄酮得率为0.89%。流动相:乙腈-0.5%冰醋酸溶液,梯度洗脱;检测波长:338nm;流速:1mL/min,用HPLC法测得穗花杉双黄酮含量为56%。结论:用D-101大孔树脂分离纯化岩柏草总黄酮简单可行,精制效果好。用上述色谱条件可用于穗花杉双黄酮的质量控制。
OBJECTIVE: To explore the optimum conditions for the separation and purification of total flavonoids from Kobusia polygala with D-101 macroporous resin. Methods: The total flavonoids were determined by ultraviolet spectrophotometry. Influencing factors such as flavonoid yield and purity, loading mass concentration, adsorption flow rate and ethanol volume fraction were used as the influencing factors to determine the total flavonoids Technology, and HPLC determination of Flavonoids in Flavonoids. Results: The macroporous resin had a good separation effect on the total flavonoids of A. japonica. The optimum conditions were as follows: loading mass concentration 8.5mg / mL, adsorption flow rate 2mL / min, ethanol volume fraction 70%, total flavonoids The rate is 0.89%. Mobile phase: acetonitrile-0.5% glacial acetic acid solution, gradient elution; detection wavelength: 338nm; flow rate: 1mL / min. Conclusion: The separation and purification of the total flavonoids of kobmats by D-101 macroporous resin is simple and feasible and the purification effect is good. The above chromatographic conditions can be used for the quality control of Fraxinus mandshurica flavonoids.