In this paper, a chromatographic method for the determination of volatile organic compounds (VOCs) as a tool for Gram-positive and Gram-negative bacterial strains identification was described. Direct Headspace gas chromatography coupled with mass spectrometry (Headspace-GC-MS) has been used for the qualitative determination of VOCs emitted by certain bacterial strains (Salmonella Typhimurium, Escherichia coli, Serratia rubidae, Staphyloccocus aureus, Enterococcus casseliflavus, and Enterococcus faecalis). In the first stage of the study, pathogenic bacteria were cultured in broth-specific liquid culture medium directly (lauryl sulfate broth) into headspace vials. Subsequently, the VOC emissions were analyzed by airborne screening in the headspace vials. Thus, the objective was to analyze all the resulting volatile organic compounds in order to select only those compounds which can be exclusively associated with specific pathogenic bacteria. Qualitative analysis by Headspace-GC-MS has proven to be a non-invasive, accurate, and fast method for identifying certain bacterial strains, based on VOCs emission. A considerable number of volatile organic compounds have been determined in the headspace air, a significant difference being observed between the VOCs emitted by bacterial cultures compared to the culture medium but also between the types of bacterial cultures themselves. The study presents preliminary results, which prove that the identification of the studied pathogenic bacteria is possible, based on the determination of certain types of VOCs in the headspace air of these cultures. This method can be used successfully for the rapid identification of bacterial culture types compared to classical methods.