The work shows a complex approach to analyzing amino acids using carbon sorbents with different surface areas, including molecular-statistical calculations, high-performance liquid chromatography, and mass spectrometry with matrix-activated and surface-activated laser desorption/ionization (MALDI, SALDI). An analysis of the thermodynamic adsorption characteristics (TACs) of a series of proteinogenic amino acids and their isomers on graphitized thermal carbon black (GTCB) is carried out. A satisfactory correlation is established between experimental characteristics of amino-acid retention on the Hypercarb carbon sorbent and TACs of these particles calculated using the molecular statistics method. It is found in the identification of amino acids using the MALDI method in the chromato-mass-spectrometric modification in the off-line mode that the application of carbon sorbents as a support results in an increase in the intensity of the registered protonated molecular ions and adducts with alkali cations compared to a conventional experimental technique on a stainless-steel surface.