In this study, the changes caused by variation of altitude to the essential oils (EOs), fatty acid methyl esters (FAMEs), and antimicrobial activities of Primula vulgaris Huds. subsp. vulgaris (Pvv) and P. vulgaris Huds. subsp. sibthorpii (Hoffmanns) W.W.Sm. & Forrest (Pvs)) grown in Turkey were investigated. Major fluctuations in the composition of Pvv and Pvs oils included methyl-4-methoxy salicylate (4.5-35.3%; Pvv and 3.2-37.2%; Pvs), (Z,Z,Z)-7,10,13-hexadecatrienal (5.1-21.8%; Pvv and 4.4-15.2%; Pvs) and flavone (5.5-14.9%; Pvv and 1.6-18.0%; Pvs). Fatty acid profile (C-60-C-260) changes were noted in Pvv and Pvs. Methyl hexadecanoate (2.4-9.3%) and methyl octadecanoate (1.0-4.7%) were present in all the FAME samples of the plants. The antimicrobial activity of the EOs of Pvv and Pvs were tested against nine bacterial species, which showed activity against Mycobacterium smegmatis with minimum inhibitory concentrations (MIC) varying from 8.5 to 59.2 mu g/mL in all samples, respectively, depending on the altitude at which the oils were obtained.