梯度洗脱HPLC法测定抗栓保心片中芍药苷和丹酚酸B的含量

HPLC determination of salvianolic acid B and paeoniflorin in Kangshuanbaoxin tablets

目的:建立高效液相色谱同时测定抗栓保心片中芍药苷和丹酚酸B含量的方法。方法:采用Agilent ZORBAX EclipseSB-C18(4.6mm×150mm,5μm)色谱柱;流动相A为5%甲醇(含0.5%磷酸),流动相B为甲醇,梯度洗脱0～15min,A80%;15～40min,A65%;流速为1mL·min-1;检测波长为230nm。结果:芍药苷和丹酚酸B保留时间分别约为11和29min,与各自相邻峰的分离度均大于1.5。以峰面积对进样浓度(μg·mL-1)线性回归,芍药苷回归方程:Y=0.07750X+1.334,r=0.9999,线性范围10.90～218.0μg·mL-1。丹酚酸B回归方程:Y=0.05373X+0.4981,r=0.9999,线性范围9.640～192.8μg·mL-1。芍药苷和丹酚酸B的回收率分别为100.4%和97.8%,RSD分别为1.7%和1.6%。结论:本方法操作简便,测定结果准确,重复性好,可用于抗栓保心片中芍药苷和丹酚酸B的含量测定。

Objective：To establish an HPLC quantitative method for the determination of salvianolic acid B and paeoniflorin in Kangshuanbaoxin tablets simultaneously. Methods：The chromatographic conditions include Agilent ZORBAX Eclipse SB - C18 （4.6 mm× 150 mm,5 μm） column, 5% methanol（ include 0. 5% phosphoric acid） as mobile phase A,methanol as mobile phase B. The flow rate was 1 mL·min^-1 ,gradient elution：0 - 15 min,A： 80% ,15 -40 min, A 65% ;and monitored at 230 nm. Results：The retention time of paeoniflorin and salvianolic acid B was about 11 min and 29 rain respectively. The resolution was more than 1.5. The regress equation for salvianolic acid B was Y=0. 05373X ＋0. 4981, r =0. 9999,and the linear range was 9. 0640 - 192.8μg·mL^-1 . Paeoniflorin was Y=0. 07750X ＋ 1. 344,r =0. 9999,and the linear range was 10.90 -218.0 μg·mL^-1 . The average recoveries of salvianolic acid B and paeoniflorin were 100. 4% and 97. 8%, RSD 1.7% and 1.6%, respectively. Conclusion ： This method is simple, time - saving and accurate, and can be used for routine analysis of paeoniflorin and salvianolic acid B in Kangshuanbaoxin tablets.