用HPLC法同时测定感冒安颗粒中没食子酸、绿原酸及咖啡酸的含量

Simultaneous determination of gallic acid,chlorogenic acid and caffeic acid contents in Ganmao'an granules by HPLC method

目的:建立用HPLC法同时测定感冒安颗粒中没食子酸、绿原酸及咖啡酸的含量。方法:采用Kromasil C18色谱柱(150mm×4.6mm,5μm);流动相:乙腈(A)-0.4%磷酸溶液(B),梯度洗脱:0～7.50min A相2%,7.50～7.51min A相2%→12%,7.51～25.00min A相12%;流速:0.8ml/min;柱温:室温;检测波长:272nm(没食子酸)、327nm(绿原酸和咖啡酸)。结果:没食子酸在0.27～8.64μg/ml范围内线性关系良好(r=0.999 9),平均回收率为100.14%,RSD为1.66%(n=9);绿原酸在1.00～32.00μg/ml范围内线性关系良好(r=0.999 7),平均回收率为98.10%,RSD为2.38%(n=9);咖啡酸在0.30～9.60μg/ml范围内线性关系良好(r=0.999 8),平均回收率为100.48%,RSD为2.28%(n=9)。结论:该方法准确、灵敏,专属性强,重现性好,对感冒安颗粒质量控制标准的提高具有参考意义。

Objective： To establish a HPLC method for simultaneous determination of gallic acid,chlorogenic acid and caffeic acid contents in Ganmao＇an granules.Methods： The separation column of Kromasil C18（150 mm×4.6 mm,5 μm） was used,acetonitrile（A）-0.4% phosphoric acid（B） was applied as the mobile phase with gradient elution： 0-7.50 min 2% A,7.50-7.51 min 2%→12% A,7.51-25.00 min 12% A.The flow rate was 0.8 ml/min.The column temperature was room temperature.The detection wavelengths were 272 nm for gallic acid,327 nm for chlorogenic acid and caffeic acid.Results： The linear ranges of gallic acid,chlorogenic acid and caffeic acid were 0.27-8.64 μg/ml（r=0.999 9）,1.00-32.00 μg/ml（r=0.999 7） and 0.30-9.60 μg/ml（r=0.999 8）,respectively.The average recoveries of gallic acid,chlorogenic acid and caffeic acid were 100.14%,RSD 1.66%（n=9）,98.10%,RSD 2.38%（n=9） and 100.48%,RSD 2.28%（n=9）,respectively.Conclusion： The method is accurate,sensitive,selective and reproducible,and could provide the reference for the improvement of quality control standard of Ganmao＇an granules.