An improved LC–S/MS method for the quantitation of adenosine concentration in mice brain microdialysates

► LC/MS/MS method is developed for the quantitative analysis of adenosine in dialysates. ► A column switching method is developed to reach high sensitivity. ► The method is applied to quantify adenosine in mice microdialysis samples. A sensitive liquid chromatography tandem mass spectrometry (LC–MS/MS) method for the determination of adenosine concentrations in mouse brain microdialysis samples was developed. High method sensitivity (LLOQ of 1.25fmol) was achieved by on-line switching column. A C18 was employed as enrichment column and a cyano based (CN-SB) as analytical column. The method was fully validated for its sensitivity, selectivity, matrix effect and stability. It was successfully applied to measure quantitatively adenosine in brain of freely moving mice after different stimuli.