Simultaneous determination of 12, 13-dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture by high performance liquid chromatography

A reversed phase high performance liquid chromatography (HPLC) method was established for the simultaneous determination of 12,13-dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture.A Grace Apollo C18 column (250 mm× 4.6 mm,5 μm) was used as the stationary phase and the mobile phase was composed of acetonitrile and aqueous phosphoric acid (0.2%,v/v).Gradient elution was carried out at the flow rate of 1.0 mL/min and the column temperature was 30 ℃.An ultraviolet (UV) detector was used with a selected wavelength of 240 nm.Calibration curves were linear within the concentration range of 4.6-45.75 μg/mL for 12,13-dihydroxyeuparin (r>0.9999) and 106.9-1068.9μg/mL for glycyrrhizic acid (r>0.9999),respectively.Recoveries were 102.18 % for 12,13-dihydroxyeuparin and 101.17% for glycyrrhizic acid.The method developed could be applied to the simultaneous determination of 12,13-dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture.