Simultaneous determination of 14 bioactive compounds in Samchulkunbi-tang using HPLC-DAD and LC-MS

Abstract

A simple high performance liquid chromatography-diode array detector (HPLC-DAD) method has been developed and validated for the simultaneous determination of 14 compounds (chlorogenic acid, albiflorin, paeoniflorin, hyperoside, naringin, hesperidin, ginsenoside Rg1, poncirin, glycyrrhizin, 6-gingerol, atractylenolide III, ginsenoside Rg3, magnolol and atractylenolide I) in Samchulkunbi-tang (SKT). The identification of 14 standard compounds was conducted using liquid chromatography–mass spectrometry (LC-MS). The marker compounds in SKT were separated on a Shiseido C₁₈ column at a column temperature of 30 °C. The mobile phase was composed of 0.1% trifluoroacetic acid in water and acetonitrile. The detection wavelengths were set at 200, 205, 280 and 330 nm. The developed method had good linearity (R² > 0.9996). The relative standard deviation (RSD) values of intra- and inter-day testing were less than 3%, with 90.63–109.95% accuracy. The results of the recovery test were 90.40–109.94%. In conclusion, this HPLC-DAD method has been successfully applied to the simultaneous determination of 14 compounds in SKT.