Metabolic and cannflavin profiles of germinated hemp seed (Cannabis sativa L.)

This study aimed to identify the optimal germination period for maximizing cannflavin A (CFL-A) and cannflavin B (CFL-B) production in hemp seeds. Utilizing ultra-high performance liquid chromatograph with orbitrap mass spectrometry (uHPLC Orbitrap-MS/MS), the research measured metabolomic activities, complemented by spectrophotometric analysis of antioxidant activity. The findings indicated that CFL-A and CFL-B concentrations progressively increased, reaching a peak at 72 h. Notably, the polar compounds saw the emergence of 4-hydroxybenzaldehyde, a derivative of enzymatic sugar-to-aldehyde conversion. In contrast, the non-polar fraction exhibited a decrease in cannabidiolic acid levels. The study also noted the potential formation of alpha-linolenic acid (omega-3 fatty acid) through hydrolysis, which serves as an energy source for germination. Metabolomic profiles were categorized by Agglomerative Hierarchical Clustering (AHC) and Principal Component Analysis (PCA), revealing significant changes from Day 0 to Day 1, followed by relative stability until Day 3. A substantial reduction in DPPH IC50 values was observed from 93.00 mg/kg (Day 0) to 43.18 mg/kg (Day 3), indicated stronger radical scavenging capacity. The highest FRAP and TPC value was recorded on Day 3, reflecting the accumulation of bioactive compounds, resulting in improved antioxidant activity.