基于高效液相色谱法测定普济消毒饮主要有效成分含量的方法学研究

ethodological Study on Determination of Main Active Ingredients in Pujixiaodu Decoction Based on High Performance Liquid Chromatography

目的:基于高效液相色谱法(high performance liquid chromatography,HPLC)建立普济消毒饮(Pujixiaodu decoction,PJXDD)中橙皮苷、黄芩苷、盐酸小檗碱和牛蒡苷等主要有效成分的含量测定方法,为PJXDD的质量评价提供参考。方法:采用Agilent 1260型高效液相色谱仪,色谱柱为AgiLent ZORBAX SB-C18柱(250 mm×4.6 mm,5μm),流动相为乙腈-0.1%磷酸溶液,梯度洗脱,流速为1.0 mL/min,柱温为30℃,求出橙皮苷、黄芩苷、盐酸小檗碱和牛蒡苷的回归方程,并考察测定方法的精密度、重复性、稳定性。结果:实验结果显示,橙皮苷、黄芩苷、盐酸小檗碱和牛蒡苷的回归方程分别为A=54.99ρ+4.11(r=0.9999)、A=47.25ρ-6.66(r=0.9999)、A=46.95ρ-12.40(r=0.9996)和A=173.37ρ-31.03(r=0.9999);精密度试验结果显示,橙皮苷、黄芩苷、盐酸小檗碱和牛蒡苷的A值相对标准偏差(RSD)分别为0.89%、0.22%、0.52%、0.86%;重复性试验结果显示,橙皮苷、黄芩苷、盐酸小檗碱和牛蒡苷含量的RSD分别为0.99%、0.23%、0.63%和0.89%;稳定性试验结果显示,橙皮苷、黄芩苷、盐酸小檗碱和牛蒡苷含量的RSD分别为0.86%、0.34%、0.83%和0.61%。结论:基于当前条件参数的HPLC可以较为准确的测定PJXDD中橙皮苷、黄芩苷、盐酸小檗碱和牛蒡苷等主要有效成分的含量,方法具有良好的精密度、重复性和稳定性。

Objective:To establish a method for assay of main active ingredients such as hesperidin,baicalin,berberine hydrochloride and arctiin in Pujixiaodu decoction(PJXDD)based on high performance liquid chromatography(HPLC),and provide reference for the quality evaluation of PJXDD.Methods:The Agilent 1260 HPLC system was used,the chromatographic column was AgiLent ZORBAX SB-C18 column(250 mm×4.6 mm,5μm),and the mobile phase was acetonitrile-0.1%phosphoric acid solution;a gradient elution was performed at a flow rate of 1.0 mL/min and a column temperature of 30℃,the regression equations of hesperidin,baicalin,berberine hydrochloride and arctiin were obtained,and the precision,repeatability and stability of the determination method was investigated.Results:The experimental results showed that the regression equations of hesperidin,baicalin,berberine hydrochloride and arctiin were A=54.99ρ+4.11(r=0.9999),A=47.25ρ-6.66(r=0.9999),A=46.95ρ-12.40(r=0.9996)and A=173.37ρ-31.03(r=0.9999)respectively;the results of precision test showed that the relative standard deviations(RSDs)of A values of hesperidin,baicalin,berberine hydrochloride and arctiin were 0.89%,0.22%,0.52%,and 0.86%respectively;the repeatability test results showed that the RSDs of the contents of hesperidin,baicalin,berberine hydrochloride,and arctiin were 0.99%,0.23%,0.63%and 0.89%respectively;the stability test results showed that the RSDs of the contents of hesperidin,baicalin,berberine hydrochloride and arctiin were 0.86%,0.34%,0.83%and 0.61%respectively.Conclusion:The HPLC based on the current conditions and parameters can be used to accurately determine the contents of main active ingredients such as hesperidin,baicalin,berberine hydrochloride and arctiin in PJXDD,and the method has good precision,repeatability and stability.