Rapid and sensitive liquid chromatography with tandem mass spectrometry method for the simultaneous determination of 11 major components of Yuanhu-Baizhi herb-pair in rat perfusion fluids

A rapid and sensitive liquid chromatography with tandem mass spectrometry method has been developed and validated for the simultaneous quantification of 11 major compounds in Yuanhu-Bazhi herb-pair from a rat intestinal perfusion experiment, including eight alkaloids and three coumarins using diazepam as the internal standard. The analysis was performed on an Agilent Poroshell 120EC-C18 column using a gradient elution of acetonitrile/H2O with 0.0015% formic acid and 2 mM ammonium formate buffer additive (pH 4.2). All compounds were detected by tandem mass spectrometry in positive ionization mode and quantified using multiple reaction monitoring mode. Good linearity in the related range of each compound was presented. The validation data, such as accuracy and precision, were within the required limits. The lower limit of quantification of each analyte was less than 10 ng/mL. Intra-day and inter-day precisions were less than 12.84%. The relative errors of accuracy were in the range of -13.31 to 13.19%. The recoveries of analytes were greater than 84.08%. The method has been successfully applied to an intestinal absorption study of major components following intestinal perfusion of Yuanhu-Bazhi herb-pair extract to rats.