Simultaneous determination of six active components in traditional herbal medicine ‘Oyaksungisan’ by HPLC–DAD

Abstract

In this study, an effective high performance liquid chromatography–diode array detector (HPLC–DAD) method was established for simultaneous determination of six marker compounds, ephedrine hydrochloride, 6-gingerol, glycyrrhizin, hesperidin, imperatorin and ferulic acid, in a Korean traditional prescription, Oyaksungisan, which is used for hemiplegia, arthralgia and paralysis. The six marker compounds of Oyaksungisan were separated on a LUNA C18 column (S-5 μm, 4.6 mm I.D. × 250 mm) at a column temperature of 35°C. The gradient elution was composed of water with 0.1% trifluoroacetic acid and methanol. The detection UV wavelengths were set at 207, 250, 280 and 320 nm. Calibration curves for the six compounds showed good linear regressions (R ² > 0.9999). The limits of detection and limits of quantification were within the ranges 0.003–0.01 and 0.01–0.04 μg/ml, respectively. The relative standard deviation (RSD) values of intra- and inter-day testing were within the ranges 0.10–1.82 and 0.04–1.59%, respectively. The results of the recovery test were 95.05–104.27% with a RSD value of 0.11–1.85%. In conclusion, the simultaneous determination method developed was useful in the quality evaluation of Oyaksungisan.