Simultaneous Determination of Chlorogenic Acid Forsythin and Baicalin In Shuanghuanglian Koufuye by HPLC

Aim To establish an HPLC quantitative method for the determination of chlorogenic acid,forsythin and baicalin in Shuanghuanglian Koufuye.Methods The chromatographic conditions include column C18(Agilent ZORBAX Eclipse XDB-C18,4.6mm×250mm,5μm),5% methanol(include 0.5% phosphorc acid) as mobile phase A,methanol as mobile phase B.The flow rate was 1.0mL/min,gradient elution:0～35min,A:95%～35%,B:5%～65%;and monitored at 324nm(0～16min),278nm(16～35min).Results The retention time of chlorogenic acid,forsythin and baicalin was 11.3min,26.6min and 28.0min respectively.The resolution was above 1.5.For chlorgenic acid the regress equation was Y=0.035 49X+0.058 00,r=0.999 9,and the linear range was 14.08～56.32μg·mL-1.For forsythin it was Y=0.177 0X-0.025 44,r=0.999 9,and the linear range was 14.34～57.34μg·mL-1.For baicalin it was Y=0.030 69X-0.798 1,r=0.999 9 and the linear range was 61.32～245.3μg·mL-1.The average recovery of chlorogenic acid,forsythin and baicalin was 99.7%,98.3% and 97.8%;RSD 0.33%,1.9% and 0.55% respectively.Conclusion This method conforms to the method of determination specified in CP 2005,and is simple,time-saving and accurate.It can be used for routine analysis of chlorogenic acid,forsythin and baicalin in Shuanghuanglian Koufuye simultaneously.