Determination of calycosin-7-O-β-D-glucoside and formononetin in crude drugs and slices of Astragalus from different habitats and gathering periods

Objective:To establish an HPLC method for the determination of calycosin-7-O-β-D-glucoside and formononetin in crude drugs and slices of Astragalus from different habitat and gathering periods.Methods: ZORBAX Eclipse XDB-C18(250 mm×4.6 mm,5 μm) was adoped with a mobile phase of acetonitrile-water,the separation was carried out with a gradient program at a flow rate of 1 mL·min-1,column temperature was 35 ℃ and detection wavelength was 260 nm.Results: Calycosin-7-O-β-D-glucoside and formononetin showed good linearity in the ranges of 0.1272-12.72 μg(r=0.9999) and 0.003344-2.508 μg(r=0.9999) with average recoveries of 98.9% and 99.0%,respectively.The contents of calycosin-7-O-β-D-glucoside and formononetin in crude drugs and slices of Astragalus from different habitat marked difference,and the contents of calycosin-7-O-β-D-glucoside and formononetin grown for six years was the highest.Conclusion: The method is simple,rapid for determination the contents of calycosin-7-O-β-D-glucoside and formononetin in Astragali Radix,and can be used as a basis for finding better habitat and gathering periods of Astragali Radix.