Industrial hemp (Cannabis sativa L.; Cannabaceae), traditionally cultivated for fiber, also represents a valuable source of nutrient-rich seed oil. In this study, cold-pressed hemp seed oils from three Romanian varieties (Teodora, Silvana, and Armanca) were evaluated for their fatty acid composition, minor bioactive constituents, antioxidant activity, growth-promoting property toward probiotic strains in vitro, and molecular docking interactions with probiotic targets. Gas chromatography revealed a fatty acid profile dominated by linoleic (49.4-51.9%), oleic (16.3-22.8%), and α-linolenic acids (9.8-14.4%), resulting in favorable PUFA/SFA ratios (5.17-6.39) and ω-6/ω-3 ratios (3.93-5.53). The oils also contained phenolics (118-160 mg GAE/kg), chlorophylls (6.18-8.31 mg/kg), and carotenoids (2.58-3.37 mg/kg), which contributed to their antioxidant activity (DPPH inhibition 35.92 µM TE/100 g-43.37 µM TE/100 g). Broth microdilution assays against Lacticaseibacillus rhamnosus GG, L. paracasei ATCC BAA-52, and L. acidophilus ATCC 4356 demonstrated strain- and dose-dependent potential to promote probiotic growth under in vitro conditions. While L. rhamnosus and L. paracasei were inhibited at low concentrations and only mildly stimulated at higher levels, L. acidophilus showed robust growth promotion, reaching +54.7% effect and CP = 1.55 with Teodora oil at 16 mg/mL. Molecular docking highlighted strong binding affinities of γ-linolenic and linoleic acids with key metabolic enzymes involved in probiotic metabolism (hydratase, enolase, glyceraldehyde-3-phosphate dehydrogenase, ribonucleoside hydrolase), forming stable hydrophilic and hydrophobic interactions which are explored in defining the stability of the ligand-protein complexes. These results indicate that both major fatty acids and minor bioactive constituents contribute to the nutritional and antioxidant value of Romanian hemp seed oils and reveal a potential to promote probiotic growth under in vitro conditions, as supported by complementary in silico evidence.

Chemical Profile, Bioactive Constituents and In Vitro Growth Stimulation Properties of Cold-Pressed Hemp Seed Oils from Romanian Varieties: In Vitro and In Silico Evaluation (2025)