Simultaneous determination of eight marker compounds in the traditional herbal medicine, Sipjundaebo-tang by HPLC-DAD.

Sipjundaebo-tang, known as a traditional herbal medicine, has been used in the treatment of anemia, inflammation, and tumor. For simultaneous determination of eight components, namely 5-HMF, paeoniflorin, ferulic acid, cinnamaldehyde, decursinol, 6-gingerol, decursin, and glycyrrhizin in Sipjundaebo-tang, a high performance liquid chromatography-diode array detector (HPLC-DAD) method was established. In order to develop and validate this HPLC-DAD method, C(18) column (S-5 μm, 4.6 × 250 mm) was used with gradient mobile phase at the column temperature of 35°C. The mobile phase was consisted of water and methanol containing 0.1% trifluoroacetic acid. UV wavelength was set at 230, 254, 280, and 300 nm. Validation of the analytical method was evaluated by linearity, precision, and accuracy test. Calibration curve for eight marker compounds showed good linearity with R(2) > 0.9994. Limits of detection and Limits of quantification ranged from 0.01 μg/mL to 0.13 μg/mL and 0.03 μg/mL to 0.41 μg/mL, respectively. The relative standard deviations (RSDs) value of precision test, intra-day and inter-day test, was less than 1.14% and 2.54%, respectively. The results of accuracy test were varied from 98.31% to 104.88% with RSD < 2%. This developed simultaneous determination method was efficient to the quality control of Sipjundaebo-tang.