Separation and Determination of Five Flavone Compounds by RP-HPLC

A reversed-phase high performance liquid chromatography(RP-HPLC) method for the separation and dertermination of the five flavonoids(rutin,morin,quercetin,luteolin and genistein) was developed.The column was 4.6 mm×150 mm i.d.,packed with Zorbax Eclipse XPB-C8(5 mm).The mobile phase was a mixture of MeOH-H2O(50∶50,V/V).The flow rate was 0.8 mL/min.UV detection was performed at 260 nm.External standard was used and the calibration curves showed good linearity over the range of 0.04～20 mg/mL(R=0.999 1),0.03～15 mg/mL(R=0.998 6),0.02～20 mg/mL(R=0.999 0),0.03～15 mg/mL(R=0.998 7)and 0.02～16 mg/mL(R=0.9989)for quercetin,rutin,morin,luteolin and genistein,respectively.Finally,this method was applied for separating and determining quercetin,rutin and genistein in Japanese pagodatree(Sophora japonica).