"Simultaneous determination of vitexin-2""-O-glucoside,vitexin-2""-O-rhamnoside, rutin, vitexin and hyperoside by HPLC" / 药物分析学报

ABSTRACT

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 μm) with the isocratic mobile phase consisting of tetrahydrofuran/acetonitrile/0.05％ phosphoric acid solution (20377, v/v/v), and the flow rate was set at 1.0 mL/min. UV detection was carried out at a wavelength of 360 nm 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-rhamnoside, 1.64-82.00 μg/mL for rutin, 1.74-87.00 μg/mL for vitexin, and 1.41-70.60 μg/mL for hyperoside with the correlation coefficient for each analyte more than 0.998.The limit of detection (LOD) and limit of qnantitation (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. Lntra- 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.