The essential oils from theCentaureagenus are well known for their pharmacological properties. The most abundant and dominant chemical components inCentaureaessential oils areß-caryophyllene, hexadecanoic acid, spathulenol, pentacosane, caryophyllene oxide, and phytol. However, whether these dominant components are the key drivers for observed antimicrobial activity remains unclear. Thus, the aim of this study was dual. Here we provide comprehensive, literature-based data to correlate the chemical compounds inCentaureaessential oils with the tested antimicrobial activity. Secondly, we characterized the essential oil ofCentaurea triumfettiiAll. squarrose knapweed using coupled system gas chromatography–mass spectrometry and tested its phytochemicals for antimicrobial activity againstE. coliandS. epidermisusing disc diffusion assay and monitoring their growth in Muller Hinton broth. The most abundant compounds inC. triumfettiiessential oil were hexadecanoic acid (11.1%), spathulenol (10.8%), longifolene (8.8%), germacrene D (8.4%), aromadendrene oxide (6.0%) and linoleic acid (5.3%). Based on our analysis of literature data from otherCentaureaessential oils, they were positively correlated with antimicrobial activity. Using an agar disk diffusion method, tested chemical constituents did not show experimental evidence to support this positive correlation to antimicrobial activity when we tested them as pure components. The antibacterial effect of essential oil constituents may be related to a complex synergistic, rather than a single component as suggested by performed network pharmacology analysis, underlying the theoretical interactions between the essential oil phytochemicals listed as potentially responsible for antimicrobial activity and should be confirmed in further in-depth studies. This is the first report on the comparative analysis ofCentaureaessential oils with good antimicrobial activity, as well as the first analysis of chemical components of the essential oil fromC. triumfettiiand the first report of antimicrobial activity of the representative, pure components: aromadendrene, germacrene D, spathulenol, longifolene, and the mixture of selected chemical compounds. This work contributes to the body of knowledge on the genusCentaureaandC. triumfettiispecies.