An optimized method for neurotransmitters and their metabolites analysis in mouse hypothalamus by high performance liquid chromatography–Q Exactive hybrid quadrupole-orbitrap high-resolution accurate mass spectrometry

•Established a method to detect 10 NTs and their metabolites in mouse hypothalamus.•LC–QE MS has been proved it can be applied for quantitative analysis of NTs.•Plain pretreatment was needed for bio-sample in complex matrices with this method. Neurotransmitters (NTs) and their metabolites are known to play an essential role in maintaining various physiological functions in nervous system. However, there are many difficulties in the detection of NTs together with their metabolites in biological samples. A new method for NTs and their metabolites detection by high performance liquid chromatography coupled with Q Exactive hybrid quadruple-orbitrap high-resolution accurate mass spectrometry (HPLC–HRMS) was established in this paper. This method was a great development of the applying of Q Exactive MS in the quantitative analysis. This method enabled a rapid quantification of ten compounds within 18min. Good linearity was obtained with a correlation coefficient above 0.99. The concentration range of the limit of detection (LOD) and the limit of quantitation (LOQ) level were 0.0008–0.05nmol/mL and 0.002–25.0nmol/mL respectively. Precisions (relative standard deviation, RSD) of this method were at 0.36–12.70%. Recovery ranges were between 81.83% and 118.04%. Concentrations of these compounds in mouse hypothalamus were detected by Q Exactive LC–MS technology with this method.