RP-HPLC法测定桃红四物汤提取物中9种水溶性活性成分的含量

Determination of Nine Kinds of Water-Soluble Active Ingredients from Tao Hong Siwu Decoction by RP-HPLC

目的建立RP-HPLC法同时测定桃红四物汤中苦杏仁苷、羟基红花黄色素A、梓醇、地黄苷A、阿魏酸、芍药苷、原儿茶酸、绿原酸、没食子酸9种水溶性活性成分的含量测定方法。方法采用Hypersil GOLD AQ(4.6 mm×250 mm,5μm)色谱柱;乙腈-水(0.05%H_3PO_4)为流动相,梯度洗脱;流速:1 mL·min^(-1);柱温:30℃;检测波长:215 nm。结果桃红四物汤中苦杏仁苷、羟基红花黄色素A、梓醇、地黄苷A、阿魏酸、芍药苷、原儿茶酸、绿原酸、没食子酸分别在6.47~129.40,4.01~80.19,2.51~50.16,3.03~60.66,1.05~21.00,5.03~100.50,0.90~18.02,0.46~9.12,0.32~6.30μg·mL^(-1)范围内浓度与峰面积呈良好线性关系,相关系数分别为0.999 0,0.999 7,0.999 5,0.999 6,0.999 8,0.999 9,0.999 8,0.999 4,0.999 6。结论该方法准确、可靠、快速,可用于桃红四物汤中9种水溶性活性成分的质量控制。

Objective To establish a RP-HPLC method for the content determination of 9 kinds of water-soluble ingredients （amygdalin, hydroxysaffloryellow A, catalpol, rehmaionoside A, ferulic acid, paeoniflorin, protocatechuic acid, chlorogenic acid and gallic acid） from Too Hong Siwu Decoction by RP-HPLC. Methods The Hypersil GO1,D AQ chromatographic column （5μm, 4.6 mm ×250 ram） was used. The 0.05 % phosphoric acid solution and acetonitrile were used as the mobile phase to separate the 9 kinds of ingredients by gradient elution. The flow rate was 1 mL·min^-1. The column temperature was set at 30℃, and the detection wavelength was set at 215 nm. Results Under the above chromatographic conditions, good linear relationship was shown between peak area and the concentration of amygdalin, hydroxysafflor yellow A, catalpol, rehmaionoside A, ferulic acid, paeoniflorin, protocatechuic acid, chlorogenic acid and gallic acid from Too Hong Siwu Decoction in the range of 6.47-129.40, 4.01-80.19, 2.51- 50.16, 3.03-60.66, 1.05-21.00, 5.03-100.50, 0.90~18.02, 0.46-9.12, 0.32-6.30μg·mL^-1 respectively. The correlation coefficient was 0.999 0, 0.999 7, 0.999 5, 0.999 6, 0.999 8, 0.999 9, 0.999 8, 0.999 4, 0.999 6 respectively. Conclusion This method is accurate, reliable and rapid, and can be used for the quality control of 9 kinds of water-soluble ingredients from Too Hong Siwu Decoction.