Quantitative analysis of flavonoids, alkaloids and saponins of Banxia Xiexin decoction using ultra-high performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry

Abstract

Banxia Xiexin decoction (BXD) is an effective Chinese Medicinal Prescription in treating gastroenteritis diseases. In this study an ultra-high performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) method was developed to separate and determine 18 major active ingredients of BXD in order to guarantee quality. The separation of ten flavonoids, four alkaloids and four saponins was accomplished on an Acquity BEH C18 (2.1mm×100mm, 1.7μm) column using gradient elution with 0.1% (v/v) formic acid water (A) and 0.1% (v/v) formic acid in methanol (B). All the analytes were detected in positive electrospray ionization tandem mass spectrometry by selective reaction monitoring (SRM) mode. A good linear regression relationship for each analyte was obtained over the range from 2.41–438ng/ml to 20.75–4150ng/ml. The precision was evaluated by intra- and inter-day assays with relative standard deviation (RSD) less than 7.7%. The recovery measured at three concentration levels varied from 92.4% to 107.8%. The method sensitivity expressed as LOQ was typically 0.97–4.15ng/ml. The assay was successfully applied for determination of the 18 bioactive compounds in BXD. The results indicated that the new UPLC–MS/MS method was rapid and accurate, and could be reliably utilized as a quality control method for BXD.