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The aim of this study was the characterization and in vitro evaluation of some of the biological activities of honey samples from Algerian flora. Physicochemical analyses were performed to evaluate the quality of the honey samples. Total phenols, flavonoid content, and free radical scavenging activity of 2,2-diphenyl-1-picrylhydrazyl (DPPH) were measured as antioxidant determinants. Finally, in vitro antimicrobial tests were carried out to evaluate the inhibitory capacity against certain microorganisms (gram-positive and gram-negative bacteria and a yeast species). According to the results, the majority of the samples met the appropriate standards for good quality. Their total phenolic content was average, while the flavonoid content was considerable. The DPPH antiradical test gave positive and strong results, with the darker colored honey and even citrus honey showing the best activity. The antimicrobial results showed significant capabilities against all bacteria studied. Only a honey sample labeled “Sulla honey” was able to inhibit the spread of the yeast Candida albicans. Statistical analyses revealed that the differences in all results were significant between samples, confirming the influence of botanical and geographical origin.