The objective of the research: To define the optimal conditions for isolation of cannabinoids and terpenes from industrial hemp inflorescences by applying "green" supercritical carbon dioxide extraction (sCO2); to quantify the obtained extracts on the content of cannabinoids and terpenes; and to determine the antioxidant and antimicrobial activities of sCO2 extracts. Material and methods: Dried inflorescences of industrial hemp (Cannabis sativa L.) of Croatia-grown Futura 75 variety were used for the research. Extraction of target bioactive compounds using sCO2 was carried out. The experimental matrix was made according to the central composite design, and the analysis of variance was used to assess the degree of accuracy of the applied methodology. Identification and quantification of volatile compounds of sCO2 extracts was determined by gas chromatography-mass spectrometry combined system (GC-MS), while cannabinoids were determined by high-performance liquid chromatography (HPLC-DAD). The antioxidant properties of the obtained extracts were determined by a DPPH scavenging assay, while antibacterial activity was tested by a microdilution method for the determination of minimal inhibitory concentrations. Results: Analysis with gas chromatography and mass spectrometry (GC-MS) showed β-myrcene chemotype in sCO2 extracts, with a high content of α-pinene and β-pinene and a number of other monoterpenes in smaller proportions, while the most dominant sesquiterpene was β-caryophyllene. The main cannabinoids of sCO2 extracts were CBDA and CBD, followed by less common CBCA, CBC, THCA, THC, CBGA, CBG, and CBDVA, while CBN and THCVA were not detected. sCO2 extract obtained at a pressure of 320 bar and a temperature of 40 °C, with the highest CBDA content, exhibited the best antioxidant properties. All extracts exhibited good antibacterial effect against E. coli, P. aeruginosa, B. subtilis and S. aureus. sCO2 extract with the highest content of CBD, which also contains a high content of α-pinene, β-pinene, β-myrcene and limonene, showed the best antibacterial activity. Optimal conditions have been established for the sCO2 extraction of cannabinoids and terpenes from industrial hemp. The temperature of 60 °C proved to be optimal for all studied responses, while pressure had a different effect depending on the compounds targeted. For monoterpenes, lower pressure was optimal, while higher pressure was suitable for the extraction of sesquiterpenes. The application of higher pressures was also desirable for CBD extraction. By using decarboxylation as a pretreatment of plant material before sCO2 extraction, extracts with a high yield of CBD were obtained. Conclusion: sCO2 extraction process for obtaining industrial hemp extracts with antioxidant and antibacterial activities and with the potential of topical application for medical and cosmetic purposes has been optimized. The provided research confirmed the selectivity of the applied technique for the extraction of targeted terpenes and cannabinoids from industrial hemp inflorescences.