황련해독탕 중 3종 생리활성 물질의 HPLC-DAD 동시 정량분석법 확립

Simultaneous Analysis of three Marker Components in Hwangryunhaedok-tang by HPLC-DAD

In this study, a high performance liquid chromatography-diode array detector method was established, for simultaneous determination of three compounds, berberine, palmatine and geniposide in Hwangryunhaedok-tang, To develop and validate method, C18 column (5 μm, 4.6 mm×250 mm) was used with gradient mobile phase, water containing 0.1% trifluoroacetic acid (TFA) and MeOH at the column temperature of 30oC. UV wavelength was set at 230 and 280 nm. Validation of the chromatography method was evaluated by linearity, precision and accuracy test. Calibration curve of standard components showed good linearity (R2>0.9999). The limits of detection (LOD) and limits of quantification (LOQ) varied from 0.05 to 0.17 μg/ml and 0.15 to 0.53 μg/ml, respectively. The relative standard deviations (RSDs) data of intra-day and inter-day test were in less than 2.99% and 1.90%, respectively. The results of the accuracy test were in the range of 98.36 to 102.52% with RSDs values 0.32 to 1.98%. The results of validation indicated that this method was a very accurate and sensitive assay.