Development and validation of a HPLC-UV-ESI-MS method for the simultaneous quantitation of ten bioactive compounds in Dahuang Fuzi Tang

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.