Origanum onites L. of Lamiaceae, is used as a culinary spice and herbal tea; also for its antioxidant, antimicrobial effects among other activities and effects. This research aims to examine the in vitro antimicrobial and antioxidant activities of the infusions and the essential oils (EOs) obtained using different conditions from the aerial parts of the processed and raw of O. onites herbal material. The EOs from O. onites were distilled at different times and characterized both by GC-FID and GC/MS systems. The resulting main compounds were identified as carvacrol (65.5–91 %), linalool (0.3–17.9 %), γ-terpinene (2.8–4.6 %), and thymol (0.5–1.5 %), respectively. In addition, Origanum infusions were prepared at four different time intervals, where the volatile compounds of the infusions were analysed using the HS-SPME-GC/MS system. Statistical Principal Component (PCA) and Hierarchical Cluster Analyses (HCA) tools were used to demonstrate the composition variations on different times for O. onites infusion and EO samples.