In the present study, the processing, characterization, and assessment of novel non-alcoholic and sugar-free drinks based on bioactive extracts from valuable natural sources, such as green tea enriched with Chios mastiha, are considered. Currently, the transition towards the consumption of healthy and sustainable food and beverages promoting human health and well-being is strongly encouraged and biologically active compounds from natural resources have a broad range of ap-plications in this sector. In this context, three beverages (all non-alcoholic, non-carbonated, and sugar-free) were created, including extracts of green tea with Chios mastiha, matcha green tea with Chios mastiha and louisa green tea with Chios mastiha, and an evaluation of their biological potential was performed. Specifically, an analysis of water, extracts, and additives for the beverage production was carried out. Microbiological and nutritional value determination was also conducted in samples of the three products. According to the experimental results, the novel health beverage produced from green tea enriched with Chios Mastiha extracts was found to have improved organoleptic characteristics and was microbiologically stable and safe for a period of 180 days from the production date at 25 °C. It is also considered stable and safe for 3 days after production, even if it remains open at 25 °C. In view of a possible scale-up of this application, safety, and preservation control should continue for at least 540 days from the date of production. In conclusion, the current research findings support the development of a novel non-alcoholic sugar-free health drink based on bioactive extracts from green tea enriched with Chios mastiha, to contribute to maintaining human health and also to strengthen the economy.