In this study, four samples of Tunisian propolis were analyzed for their volatile compounds. Static Headspace technique coupled with gas chromatography-mass spectrometry (GC/MS) was used for the volatile profile characterization. Statistical investigation of the samples was made applying hierarchical clustering method, K-Means cluster and principal component analysis. Results showed that 47, 36, 30 and 24 different compounds were identified, respectively, in the samples from Zouarine, Zelligua, Bizerte and Beni Khalled. Propolis volatiles were dominated by monoterpene hydrocarbons. α-pinene was the major compound representing 81.14%, 82.67% and 90.74%, respectively, of the total propolis volatiles collected from Zelligua, Beni Khalled and Zouarine and only 45.22% of the sample from Bizerte which had a very different composition. The in vitro antifungal activity of the volatiles from all samples against Candida albicans was also assayed and reported.