Simultaneous Determination of Mangiferin,Calycosin-7-O-β-D-glucoside and Calycosin in Anemarrhenae Rhizoma-Astragali Radix by HPLC

Objective: A reversed-phase high performance liquid chromatographic method(RP-HPLC) was developed for the simultaneous analysis of mangiferin,calycosin-7-O-β-D-glucoside and calycosin in Anemarrhenae Rhizoma-Astragali Radix.Method:HPLC determination was performed on a Phenomenex Luna C18(4.8 mm×250 mm,5 μm) and detected at 250 nm.The mobile phase was consisted of acetonitrile and 0.2% phosphate solution with gradient elution.Flow-rate was 1.0 mL·min-1,and column temperature was 30 ℃.Result: The method was proved to be linear in the ranges of 36.733-550.999 ng,33.376-166.88 ng and 44.544-222.72 ng for mangiferin calycosin-7-O-β-D-glucoside and calycosin respectively.The average recoveries were 102.47%,102.38% and 100.20% respectively.Conclusion: The results showed that the method is convenient,accurate and reliable for the simultaneous determination of mangiferin,calycosin-7-O-β-D-glucoside and calycosin in Anemarrhenae Rhizoma-Astragali Radix.