Simultaneous determination of five bioactive constituents in the traditional chinese medicinal preparation refined Xue-Fu-Zhu-Yu-Tang by high-performance liquid chromatography with diode-array UV detection

A high-performance liquid chromatographic method for the simultaneous determination of oxypaeoniflorin, paeoniflorin, ferric acid, naringin, and neohesperidin in a traditional Chinese medicinal preparation, refined xue-fu-zhu-yu-tang, was developed. Separations were carried out with a ZORBAX SB-C-18,8 column along with an Agilent C-18 precolumn by linear gradient elution using 1% acetic acid-methanol (v/v: 0 min, 80/20; 15 min, 70/30; 35 min, 60/40; 55 min, 60/40) as the mobile phase at a flow rate of 0.5 mL/min. The analytes were detected with a diode-array detector at their maximum UV wavelengths. The correlation coefficients of the calibration curve for the analytes exceeded 0.9999. The recoveries were in the range 95.95-104.42% with RSD less than 1.85%, and the precision of this method was better than 1.80%. The effects of several factors on the chromatographic behavior and sample extraction of the analytes were compared. In addition, the contents of these bioactive constituents in six different samples of this preparation were analyzed by using the optimized high-performance liquid chromatographic method.