Determination of the Major Chemical Components in Different Combinations of Dahuang Huanglian Xiexin Decoction by HPLC-DAD

ObjectiveThe aim of this study is to determine the major chemical components in different herbs and combinations of the Chinese medicine formula Dahuang Huanglian Xiexin Decoction (DHXD), which is composed of Rhizoma rhei, Rhizoma coptidis, and Radix scutellariae, with High Performance Liquid Chromatography with Diode Array Detector (HPLC-DAD) and Liquid Chromatography Tandem Mass Spectrometry (LC/MS/MS) methods.MethodsA HPLC-DAD method was established with Luna octadecyl silane (ODS) column (250 mm × 4.6 mm, 5 μm). The mobile phase was acetonitrile-0.5% triethylamine (adjusted by phosphoric acid, pH 3.5) gradient elution with the flow rate of 1.0 mL/min. The sample size was 20 μL. The chromatographic peaks of the individual herbs and the DHXD formula were identified by comparing the relative retention time and ultraviolet absorption spectrum with the reference compounds, together with LC/MS/MS.ResultsThe HPLC-DAD method was accurate and reproducible. A total of 20 chromatographic peaks were observed. Eleven peaks, including berberine from Coptis, were identified using reference compounds, and four peaks, including coptisine, were determined by LC/MS/MS.ConclusionThe HPLC-DAD method is suitable for qualitative analysis of major chemical components in DHXD. The identification of the 15 chemical components and determination of their source herbs in the formula provides new analytical evidence on the chemical basis and combinational principles and quality control of DHXD.