Development of a high performance liquid chromatography method for the quantitative determination of bioactive triterpenoids in the extracts of Antrodia camphorata

Abstract

A simple gradient high-performance liquid chromatography with diode array detection (HPLC-DAD) method was developed for the simultaneous determination of total ten triterpenoids in Antrodia camphorata (AC). Optimum separation was obtained with a J'sphere C18 column using gradient elution with 0.2% aqueous acetic acid and acetonitrile as the mobile phase, a flow rate of 0.8 mL min⁻¹, detection wavelength at 248 nm and sample volume at 5 μL. This method produced linear responses for ten triterpenoids in the concentration range of 50–800 μg mL⁻¹. This new method was validated for acceptable precision (intra- and inter-day precision was less than 2.47% and 3.94%, respectively), accuracy (recovery ranged from 96.46% to 102.37%) and sensitivity (LODs and LOQs were in the range of 15.7–50.5 μg mL⁻¹, and 47.8–194.0 μg mL⁻¹, respectively). This method has been successfully applied to the analysis of real AC samples. Quantitation was achieved by direct comparison of the peaks of AC extract of the sample to reference standards of known concentrations. The present results may be useful for evaluating the quality of the AC and its products.