Simultaneous Determination of Six Components in the Traditional Prescription 'Oryeongsan' by HPLC-DAD

Oryeongsan is one of the Chinese traditional prescriptions that exhibits biological activity against symptoms of fever, irritability, urinary difficulty and a floating pulse. A simple high performance liquid chromatography – diode array detector (HPLC-DAD) method has been developed and validated for simultaneous determination of the six components (cinnamic acid, cinnamaldehyde, eugenol, atractylenolide I, atractylenolide III, and ergosterol) in Oryeongsan. The six compounds of Oryeongsan were separated on Shishedo C₁₈ column (5 µm, 4.6 mm I.D. x 250 mm) at a column temperature of 30 °C The mobile phase was a mixture of 0.1% TFA (trifluoroacetic acid) water and acetonitrile employing gradient elution at a flow rate of 1.0mL/min. The detection wavelength was set at 205, 250, 280, and 330nm. The developed method had good linearity (R²> 0.9997) and the limits of detection (LOD) and limits of quantification (LOQ) were observed within the ranges 0.01 – 0.15 and 0.05 – 0.45 µg/mL, respectively. The relative standard deviation (RSD) values of intra- and inter-day testing were indicated that less than 3% and 90.31 – 103.31% of accuracy. The results of recovery test were 90.56 – 106.72% and RSD range were measured from 0.84 to 2.95%. In conclusion, this HPLC-DAD method has been successfully applied to the simultaneous determination of six components in Oryeongsan. Acknowledgements: This research was supported by a grant [K12050] from the Korea Institute of Oriental Medicine.