高效液相色谱法同时测定健脾益肾方水提物中松果菊苷、丹酚酸B和大黄酸含量

Simultaneously Content Determination of Echinacoside,Salvianolic Acid B and Rheic Acid in Jianpi Yishen Formula by HPLC

目的建立同时测定健脾益肾方水提物中松果菊苷、丹酚酸B和大黄酸含量的高效液相色谱法。方法色谱柱采用Inertsil ODS-SP柱(150 mm×4.6 mm,5μm),以乙腈-0.2%磷酸溶液为流动相梯度洗脱,流速为1.0 mL/min,检测波长330 nm(松果菊苷)、286 nm(丹酚酸B)、254 nm(大黄酸),进样量10μL。结果松果菊苷、丹酚酸B、大黄酸检测质量浓度的线性范围为2.50~80.0μg/mL(r=0.999 4),1.25~80.00μg/mL(r=0.999 3),1.25~40.00μg/mL(r=0.999 4);精密度、稳定性、重复性试验的RSD小于2%(n=6);平均回收率分别为100.01%,99.15%,98.95%,RSD均小于2%(n=6)。结论该方法,操作简单、重复性好、专属性强,方法可靠,可作为健脾益肾方水提物中松果菊苷、丹酚酸B、大黄酸的含量测定方法。

Objective To establish an HPLC method for the content determination of echinacoside,salvianolic acid B and rheic acid in Jianpi Yishen Formual simultaneously. Methods The Inertsil ODS-SP column（ 150 mm × 4. 6 mm, 5 μm） was adopted with acetonitrile -0. 2% phosphoric acid as mobile phase using gradient program, the flowing rate was 1. 0 mL/min, the detection wavelength was 330 nm（ echinacea）,286 nm（ salvianolic acid B）,and 254 nm（ rhein acid）,the sample size was 10 μL. Results The calibration curves of echinacoside, salvianolic acid B and rheic acid were linear from 2. 50-80. 00 μg/mL（ r = 0. 999 4）, 1. 25-80. 00 μg/mL（ r =0. 999 3）,1. 25-40. 00（ r = 0. 999 4） μg/mL,the accuracy,stability,and reproducibility of the RSD were less than 2. 00%（ n = 6）,the average recovery was 100. 01%, 99. 15%, 98. 95%, respectively, the RSD was less than 2. 00%（ n = 6）. Conclusion The method is simple,efficient,accurate and reproducible,which can be used for the content determination of echinacoside,salvianolic acid B and rheic acid in Jianpi Yishen Formula.