In this research, essential oil was obtained from the aerial parts of Prangos platychlaena Boiss. by hydrodistillation using a Clevenger-type apparatus, separated into fractions having different polarities by column chromatography. Both essential oil and the fractions were analyzed by GC-FID and GC/MS simultaneously. Nona-3,5-diyne-2-yl acetate (46%) and 3,5-nonadiyne (13.5%) were found to be the main constituents of the essential oil. While the main components of the n-hexane fraction were characterized as 3,5-nonadiyne (45.6%) and germacrene B (16.4%), the major components of the methanol fraction were found to be nona-3,5-diyne-2-yl acetate (59.6%) and 3,5-nonadiyne-2-ol (25.9%). In addition, principal multivariate statistical analyses were performed with principal component analyses and Venn diagram calculations, utilizing chemical compositions of the essential oil and the fractions. Furthermore, in vitro anti-inflammatory activities of the essential oil and the fractions were evaluated to correlate the chemical composition with the biological activity, and to the best of our knowledge, this study was performed for the first time in this aspect. LOX inhibitions of the essential oil, n-hexane, and methanol fractions were determined to be 70.98 ± 1.7, 67.10 ± 2.5, and 50.11 ± 4.8%, respectively. Preliminary initial findings of this study will be extended in the future with new biological assays.