We present here a tutorial review on the electrospray mass spectrometry technique and its applications to the study of drug-nucleic acid non-covalent complexes. Particular emphasis has been made on the basic principles of the technique, to allow even the non-specialist to design fit-for-purpose mass spectrometry experiments and interpret the results. Standard applications will be described in detail, including the determination of stoichiometries and equilibrium binding constants of non-covalent complexes, the study of binding kinetics, and the development of ligand screening assays. We also outline the potentials of more advanced and/or more recent MS-based techniques (tandem mass spectrometry, ion mobility spectrometry and gas-phase spectroscopy) for the study of the nucleic acid-ligand complexes.