Simultaneous determination of four triterpenoid saponins and two liganoids in the roots of Ilex pubescens by RP­HPLC­DAD

A reversed­phase high performance liquid chromatography (RP­HPLC) method with diode array detector (DAD) has been established to simultaneously determine six bioactive compounds in the roots of Ilex pubescens, namely tortoside A, (+)­syringaresinol, ilexsaponin B3, ilexsaponin A1, ilexsaponin B1, ilexgenin A. The RP­HPLC assay was performed on a reversed­ phase C18 column with a gradient elution. The mobile phase consisted of acetonitrile and water containing 0.1% (v/v) phosphoric acid. The flow rate was 1.0 mL/min and the detection wavelength was set at 210 nm. The six marker constituents were separated well with good linearity (r 2 >0.9996), precision, stability and repeatability. The overall recoveries were in the range of 99.00%-104.52%. Cluster analysis was employed to analyze 15 batches of samples. The result indicated this method provide an efficient way to perform quality control as well as a scientific rationale for the Geo­authentication of I. pubescens.