Development of a liquid chromatography–tandem mass spectrometry method for simultaneous determination of five isoflavonoids and seven neurochemicals in rat brain dialysate and its application to a pharmacological study

Pueraria lobata is a medicinal plant widely used in traditional Chinese medicine. The total pueraria isoflavones have demonstrated positive effect against neurological disorders. In the present study, we first develop an ultra high performance liquid chromatography and tandem mass spectrometry method to quantify the multiple active pueraria isoflavonoids and neurochemical markers in brain dialysate to provide tools for further exploring the functional mechanism of pueraria isoflavones for neuroactivities. A phenomenex Luna C18 column (50 × 2.0 mm, 5 μm) was employed with acetonitrile/0.05% formic acid in water as the mobile phase for the separation of analytes. A mass spectrometer with electrospray ionization source in positive/negative ion switching mode was used for multiple reaction monitoring of the detected compounds. The method was validated and proved acceptable. The intra- and interday precision across quality control levels was within 13.87 for all analytes, whereas the deviation of assay accuracies ranged between 0.03 and 11.53%. The method was successfully applied to a pharmacological study of pueraria isoflavones in rat brain.