高效液相色谱法同时测定双黄连口服液中绿原酸和木犀草苷的含量

Simultaneous Determination of Chlorogenic Acid and Cynaroside in Shuanghuanglian Oral Liquid by HPLC

目的建立高效液相色谱法(HPLC)同时测定双黄连口服液中绿原酸和木犀草苷含量的方法。方法采用HPLC对双黄连口服液中绿原酸和木犀草苷的含量进行检测,色谱柱为Kromasil C18柱(4.6 mm×250 mm,5μm),流动相A为0.2%磷酸水溶液,流动相B为乙腈,流速为1.0 ml/min,检测波长为350 nm。梯度洗脱程序为0~15 min:A为87%,B为13%;15~20 min:A为87%~80%,B为13%~20%;20~35 min:A为80%~77%,B为20%~23%;35~40 min:A为77%~20%,B为23%~80%。结果绿原酸的回归方程是Y=841.15X+358.35,r=0.999 5,线性关系范围为1.53~15.3μg/ml;木犀草苷的回归方程是Y=2 589.9X-1.293,r=0.999 3,线性关系范围为0.055~0.55μg/ml。绿原酸和木犀草苷的平均回收率分别为98.75%和98.99%,相对标准偏差(RSD)分别为2.06%和3.22%。结论该测定方法操作简便,结果稳定可靠,可用于同时测定双黄连口服液中绿原酸和木犀草苷的含量。

Objective To establish a High Performance Liquid Chromatography(HPLC) method for content determination of chlorogenic acid and cynaroside in Shuanghuanglian Oral Liquid. Methods Chlorogenic acid and luteolin in Shuanghuanglian Oral Liquid were determined by HPLC. Kromasil C18 column(4.6 mm×250 mm, 5 μm), elution A of 0.2% phosphoric acid and B of acetonitrile;the flow rate was 1.0 m L/min, and the detected wave length was 350 nm. The gradient elution was: 0-15 min: A was 87%, B was 13%;15-20 min: A was 87%-80%, B was 13%-20%;20-35 min: A was 80%-77%, B was 20%-23%;35-40 min: A was77%-20%, B is 23%-80%. Results The regress equation of chlorogenic acid was Y = 841.15 X+358.35, r = 0.9995 and the linear range was 1.53-15.3 μg·mL-1;the regression equation of luteolin was Y = 2 589.9 X-41.293, r = 0.9993 and the linear range was0.055-0.55 μg·mL-1. The average recovery rate of chlorogenic acid and cynaroside was 98.75% and 98.99%, respectively and RSD of 2.06% and 3.22% respectively. Conclusion The method is simple, reliable, stable, and reproducible and could simultaneously determine chlorogenic acid and cynaroside in Shuanghuanglian Oral Liquid.