Simultaneous determination of ferulic acid, hesperidin, 6-gingerol and glycyrrhizin in Insampaedok-san by HPLC coupled with diode array detection

Abstract

For the simultaneous determination of ferulic acid, hesperidin, 6-gingerol and glycyrrhizin in Insampaedok-san, one of traditional oriental medicine prescription, a high performance liquid chromatography (HPLC) analytical method was developed. The separation was achieved on a SHISEIDO C18 column (S-5 μm, 4.6 mm I.D. × 250 mm). The mobile phase consisted of water with 0.1% trifluoroacetic acid and methanol in gradient elution at a flow rate of 1.0 mL/min. The UV wavelength was set at 250, 280 and 320 nm. The method was validated for linearity, precision and recovery. All calibration curves showed good linear regression (r 2 > 0.9994) over the wide test ranges. LOD (limit of detection) and LOQ (limit of qualification) ranged from 0.02 to 0.14 μg/mL and 0.05 to 0.42 μg/mL, respectively. The relative standard deviation (RSD) of the intra- and inter-day test, precision test were within 2.26 and 2.45%, respectively, with accuracy from 95.95 to 108.22%. The recoveries of the four compounds were obtained between 94.30 and 106.34% with RSD less than 1.96%. The verified method was successfully applied to quality control of Insampaedok-san.