[A reversed-phase high performance liquid chromatographic method for the determination of pinoresinol diglucopyranoside in Eucommia Ulmoides Oliv].

A reversed-phase high performance liquid chromatographic method for the determination of (+)-pinoresinol-di-O-beta-D-glucopyranoside in the bark of Eucommia Ulmoides was established. The air-dried bark of E. Ulmoides was ground into powder and extracted continuously with methanol. The extract was evaporated under reduced pressure, then suspended in water. This suspension being subjected to column chromatography was eluted successively with water, 30%, 50% and 90% aqueous solution of ethanol. The 30% EtOH eluate was concentrated and then injected into HPLC system. The conditions of HPLC were: YWG-C18 column (250 mm x 4.6 mm i.d. 10 microns); 28%(V/V) methanol mobile phase and UV detector at 232 nm. The calibration curve was linear in the range of 0.068 microgram-0.68 microgram of(+)-pinoresinol-di-O-beta-D-glucopyranoside with correlation coefficient of 0.9999. The coefficient of variation and average recovery were 0.50%-0.74% (n = 5) and 99.22% (n = 3) respectively.