Synergistic Fermentation of Hemp Milk with Probiotics and Herbal Extracts for Enhancing the Performance of Male Athletes - Supplementary Data

This study reports the development of a functional, plant-based beverage via synergistic fermentation of hemp seed milk (Cannabis sativa L.) with Bacillus subtilis (Natto strain) and Lactobacillus plantarum (~10⁸ CFU/mL), enriched with standardized extracts of Withania somnifera (Ashwagandha) and Camellia sinensis (green tea). Fermentation was performed under controlled aerobic and anaerobic conditions at varying temperatures (30–42 °C) and durations (24–72 h) to optimize microbial viability, bioactive metabolite production, and antioxidant activity. Post-fermentation, the beverage was characterized for pH, titratable acidity, viable cell count, antioxidant capacity (DPPH and ABTS assays), total phenolic content (Folin–Ciocalteu method), γ-aminobutyric acid (colorimetric assay), short-chain fatty acids (GC-MS), and macronutrient composition using established biochemical methods. Sensory and stability studies ensured product acceptability and shelf-life. This approach allows reproducibility of the fermentation process, monitoring of functional metabolites, and evaluation of nutritional and adaptogenic benefits relevant to male athletes. No human or animal subjects were involved; therefore, ethical approval was not required. The data support the beverage’s potential as a natural, holistic functional drink with antioxidant, adaptogenic, and performance-enhancing properties.