Determination of Luteolin and Apigenin in Chrysanthemum morifolium Extract by RP-HPLC

Objective To develop a reversed-phase high performance liquid chromatographic method for the simultaneous determination of luteolin and apigenin in Chrysanthemum morifolium extract.Methods The separation was carried out on a Restek Pinnacle Ⅱ C18 column(250 mm×4.6 mm,5 μm)with MeOH∶THF∶0.3%H3PO4=42∶13∶56 as mobile phase at a flow rate of 1.0 ml/min.The absorbance was monitored at 350 nm.The volume of injection was 10 μl.Results The linearity was good within the ranges of 3.99 μg/ml～99.75 μg/ml(r=0.999 6)for luteolin and 4.095 μg/ml～102.375 μg/ml(r=0.999 6)for apigenin.The average recoveries were 105.9% and 103.7% and the relative standard deviations(RSDs)were 1.5% and 2.5% respectively.Conclusion The method used for the determination of luteolin and apigenin in the Chrysanthemum morifolium extract has the characteristics of short retention and high resolution.