杭白芍九种主要成分的高效液相色谱定量分析及提取工艺研究

Quantitative determination and optimun extraction technique of nine compounds of Paeoniae Radix Alba

目的:建立同时测定杭白芍9种主要成分没食子酸、羟基芍药苷、儿茶素、芍药内酯苷、芍药苷、五没食子酰葡萄糖、苯甲酸、苯甲酰芍药苷和丹皮酚的高效液相色谱法(HPLC),优化杭白芍的提取工艺。方法:HPLC检测采用0.1%磷酸水溶液-乙腈为流动相线性梯度洗脱,流速为0.8 mL/min,柱温为30℃,检测波长为230 nm。采用超声提取法,甲醇和乙醇为提取溶剂,选用L9(34)表分别设计三因素三水平的正交试验,考察溶剂浓度、液料比和提取时间对杭白芍9种主要成分总含量的影响。结果:所建立的HPLC法具有较高的专属性、灵敏度和准确性,可用于同时定量分析杭白芍9种主要成分;确定了杭白芍9种主要成分的最佳提取工艺,即采用70%乙醇为提取溶剂,液料比为200 mL/g,超声提取时间为30 min。结论:建立了同时测定杭白芍9种主要成分的HPLC法及其最佳工艺条件。

Objective: To establish the optimum extraction technique and high performance liquid chromatographic(HPLC)method to simultaneously quantify nine compounds of gallic acid,hydroxy-paeoniflorin,catechin,albiflorin,paeoniflorin,pentagalloylglucose,benzoic acid,benzoylpaeoniflorin and paeonol in Paeoniae Radix Alba.Methods:Linear gradient elution was applied using water containing 0.1%phosphoric acid and acetonitrile as the mobile phase with a flow rate of 0.8 mL/min,column temperature of 30℃and wavelength of 230 nm.The method of ultrasound extraction was used.Methanol and ethanol were used as extraction solvents,and three factors and three levels of orthogonal experiments was designed using L 9(34)table to investigate the effects of solvent concentration,ratio of liquid to material and extraction time on the total content of nine components of Paeoniae Radix Alba.Results:HPLC method was verified to have high specificity,sensitivity and accuracy through methodological validation,and it could be used for simultaneous quantitative analysis of nine components of Paeoniae Radix Alba.The results showed that the optimum extraction technology of nine components of Paeoniae Radix Alba was using 70%ethanol as extraction solvent,ratio of liquid to material was 200 mL/g and ultrasound extraction time was 30 min.Conclusion:HPLC method for the simultaneous determination of nine components of Paeoniae Radix Alba is established,and the optimum extraction technology is confirmed.