Abstract: Aim To establish an RP-HPLC method for simultaneous determination of 2''-O-rhamnosyl vitexin and vitexin in Chinese hawthorn leaf and its extract. Methods Chromatography was carried out on Kromasil C18 column (250 mm × 4.6 mm, 5 μm), using THF-CH3CN-H2O-H3PO4 (30:5:125:0.1) as the mobile phase at a flow rate of 1.0 mL·min^-1. The UV detection wavelength was 270 nm. Results The linear range of 2''-O-rhamnosyl vitexin and vitexin were 0.106 4 μg -2.1280 μg (r = 0.9991) and 0.1392 μg - 2.7840 μg (r = 0.9993), respectively. The average recoveries of 2''-O-rhamnosyl vitexin and vitexin in Chinese hawthorn leaf were 99.2% (RSD = 2.80%, n = 6) and 100.6% (RSD = 2.84%, n = 6), respectively. Conclusion This method is reproducible, simple, precise, and rapid for simultaneous determination of 2''-O-rhamnosyl vitexin and vitexin in Chinese hawthorn leaf and its extract, thereby providinge the basis for quality specification of Chinese hawthorn leaf and its extract.

Key words: Chinese hawthorn leaf, Chinese hawthorn leaf, 2''-O-rhamnosyl vitexin, 2''-O-rhamnosyl vitexin, vitexin, vitexin, RP-HPLC, RP-HPLC