Nicotinamide, nicotinic acid, nicotinamide mononucleotide, nicotinamide riboside and nicotinamide adenine dinucleotide represent key metabolites through which a variety of cellular processes is regulated. Here, we report a quantitative, simultaneous determination of nicotinamide mononucleotide, nicotinamide adenine dinucleotide and their pyridine precursors in several murine tissues using a high-performance liquid chromatography with tandem mass spectrometry method based on reversedphase chromatographic separation. All the analytical steps, including sample preparation and compounds extraction, reversed-phase liquid chromatographic separation and electrospray ionization mass spectrometry monitoring and identification, were optimized and validated in order to achieve low limit of quantification, high sensitivity and good robustness for metabolomic studies. In the analyzed tissues, we have quantified the compounds of interest, which were in the concentration range of picomoles/mg of wet tissue.