Simultaneous quantification and semi-quantification of amentoflavone and its metabolites in human intestinal bacteria by liquid chromatography Orbitrap high-resolution mass spectrometry

A quick, easy, effective method followed by ultra-high-pressure liquid chromatography coupled with linear ion trap-Orbitrap tandem mass spectrometry (UHPLC-LTQ-Orbitrap MS) was developed for the simultaneous identification and quantification of the metabolites produced by amentoflavone (AMF) in human intestinal bacteria from human feces. The method validated for quantification of AMF concerning precision, accuracy, recovery, matrix effect, stability and limits showed acceptable results. Compared with blank human intestinal bacteria chromatography, three metabolites were identified based on high-accuracy protonated precursors and multi-stage mass spectrometry (MSn) using the proposed strategy. At the same time, a new method was developed for semi-quantification of three metabolites. We describe the trend over 24 h of concentration-time curves for AMF and its metabolites. Moreover, the main metabolic pathway of AMF was clarified in human intestinal bacteria. The method was validated and successfully applied to the detection and quantification of AMF and its metabolites.