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A simple, precise, and rapid high-performance liquid chromatographic method was developed and validated for the simultaneous determination of vitexin-2＂-O-glucoside, vitexin-2＂-O-rhamnoside, rutin, vitexin, and hyperoside. The HPLC separation was performed using a Shim-pack VP-ODS C18 column （250 mm ~ 4.6 mm i.d., 5 ~tm） with the isocratic mobile phase consisting of tetrahydrofuran/ acetonitrile/0.05% phosphoric acid solution （20：3：77, v/v/v）, and the flow rate was set at 1.0 mL/min. UV detection was carried out at a wavelength of 360 nrn and the whole analysis took 25 min. The method was linear in the range of 4.12-206.00 μg/mL for vitexin-2＂-O-glucoside, 4.05-202.50 μg/mL for vitexin-2＂-O- rhanmoside, 1.64-82.00 pμg/mL for rutin, 1.74-87.00 gg/mL for vitexin, and 1.41-70.60 p.g/mL for hyperoside with the correlation coefficient for each analyte more than 0.998. The limit of detection （LOD） and limit of quantitation （LOQ） were 0.6 and 2 ng for vitexin-2＂-O-glucoside, 0.6 and 2 ng for vitexin-2＂-O-rhamnoside, 0.3 and 1 ng for rutin, 1 and 3 ng for vitexin, and 0.5 and 2 ng for hyperoside, respectively. Intra- and inter-day precision and accuracy （RSD） were less than 3%. The developed HPLC method was successfully applied to the analysis of five flavonoids in hawthorn leaves, hawthorn fruits, and the preparations containing hawthorn leaves or fruits.