AIM: To develop a high performance liquid chromatography(HPLC) coupled with electrospray ionization mass spectrometry(ESI-MS) and ultraviolet(UV) detector method for the acid-alkaline simultaneous determination of ten bioactive compounds, and analyze the effect of compatible medicinal plants on the concentration of components in Dahuang Fuzi Tang(DFT). METHOD: The chromatographic separation was performed on a Hypersil BDS C18 analytical column by gradient elution with acetonitrile and formate buffer(containing 0.15% formic acid, V/V) at 25 °C with a flow rate of 1.0 mL ·min–1 and UV detection at 280 nm. Four of the ten compounds in DFT were identified and their MS fragments were elucidated by HPLC-ESI-MS, and the contents of the six compounds were determined by HPLC-UV. RESULTS: All calibration curves showed good linear regression(r2 ≥ 0.9990). The limits of detection and limits of quantification were 0.021–0.155 μg·mL –1 and 0.076–0.520 μg·mL –1, respectively. Overall precision RSD(intra-day and inter-day) were less than 2.96%, and the average recoveries were 98.35%–101.45%, with RSD ranging from 1.54% to 3.01% for the analytes. CONCLUSION: The developed method can be applied for the quality control and provide analytical evidence on the chemical basis and combinational principles of DFT.
AIM: To develop a high performance liquid chromatography (HPLC) coupled with electrospray ionization mass spectrometry (ESI-MS) and ultraviolet (UV) detector method for the acid-alkaline simultaneous determination of ten bioactive compounds, and analyze the effect of compatible medicinal plants on the concentration of components in Dahuang Fuzi Tang (DFT). METHOD: The chromatographic separation was performed on a Hypersil BDS C18 analytical column by gradient elution with acetonitrile and formate buffer (containing 0.15% formic acid, V/V) at 25 ℃ with a flow rate of 1.0 mL.min^-1 and UV detection at 280 nm. Four of the ten compounds in DFT were identified and their MS fragments were elucidated by HPLC-ESI-MS, and the contents of the six compounds were determined by HPLC-UV. RESULTS: All calibration curves showed good linear regression (r^2 ≥ 0.9990). The limits of detection and limits of quantification were 0.021-0.155 μg.mL ^-1 and 0.076-0.520 μg.mL ^-1, respectively. Overall precision RSD (intra-day and inter-day) were less than 2.96%, and the average recoveries were 98.35% 101.45%, with RSD ranging from 1.54% to 3.01% for the analytes. CONCLUSION: The developed method can be applied for the quality control and provide analytical evidence on the chemical basis and combinational principles of DFT.