Simultaneous quantitation and validation of method for the quality evaluation of Eucommiae cortex by HPLC/UV

Abstract

A new HPLC/UV method has been developed for the simultaneous quantitative determination of four major components in Eucommiae cortex, namely geniposidic acid (1), geniposide (2), pinoresinol di-O-beta-d-glucopyranoside (3), and liriodendrin (4). Simultaneous separations of these four components were achieved on a J'sphere ODS C-18 column (250 x 4.6 mm, 4 A mu m). The elution was done using water with 0.1 % phosphoric acid (A) and acetonitrile with 0.1 % phosphoric acid (B) in a two-step elution of the mobile phase at a flow rate of 1.0 mL/min and a wavelength of 230 nm. The method was validated for linearity, recovery, precision, accuracy, stability and robustness. All calibration curves showed good linear regression (r (2) > 0.999) within the test ranges. This method showed good recovery and reproducibility for the quantification of these four components in 85 species of Eucommiae cortex. The intra-day and inter-day precisions were lower than 0.53 % (as a relative standard deviation, RSD) and accuracies between 93.00 and 106.28 % for all standards. The results indicate that the established HPLC/UV method is suitable for quantitation and quality evaluation of Eucommiae cortex.