Tremendous progress in biomedical sciences has been made possible in part by recent advances in bioanalytical methods, in particular biological mass spectrometry. Since the introduction of electrospray ionization mass spectrometry (ESI-MS) in 1984 and matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) in 1988, the field of bioanalytical mass spectrometry has seen rapid growth. In concert with separation techniques such as capillary electrophoresis and high performance liquid chromatography, mass spectrometry allows characterization of a large array of small organic molecules, peptides, proteins, oligonucleotides, and RNA fragments. Thus, substantially more expedient and definitive determination of molecular weight is now possible by mass spectrometric analysis. In this commentary, general descriptions of ESI- and MALDI-MS are presented. Furthermore, several recent developments and applications in addressing difficult biological problems are discussed.