HPLC determination of three polymethoxylated flavones in the fraction of orange peel

Objective :To set up a quantitative determination method of HPLC for three polymethoxylated flavones in the fraction of orange peel.Methods:The determination was carried out with Kromasil C ls column (4.6 mmx 250 mm,5 μm).The mobile phase consisted of acetonitrile -0.2% acetic acid (1:1).The flow rate was 1.0 mL· min~(-1).The detection wavelength was at 347 nm.Results:The calibration curves were linear at ranges of 0.0732 - 0.732 μg (for 3,5,6,7,8,3',4'-heptemthoxyflavone,r=0.9999),0.0654-0.654 μg (for nobiletin,r= 0.9999) and 0.07 -0.700 μg(for Natsudaidai,r=0.9999).The average recoveries (n=5) were 97.80% (RSD =1.9%),96.71%(RSD=1.2%),and 98.52%(RSD=2.1%)respectively.And the response factors of 3,5,6, 7,8,3',4' -heptemthoxyflavone,nobiletin and Natsudaidai were determined at 347 nm.They were 3283.6, 3331.8,2503.4,respectively.Conclusion :The method is simple with good reproducibility and can be used as the quality control of the polymethoxylated flavones fraction of orange peel.