Effect of solvent polarity on phenolic antioxidant extraction from Cannabis sativa L. leaves and inflorescences: mixture design optimization and HPLC-DAD/ESI-MS2 profiling.

El Maabiche O.; Benkirane C.; El Ati R.; Haddad Y.; Fauconnier M.; Allai L.; Mansouri F. · RSC advances · 2026

**Background & Methods** This study employed a mixture design approach to optimize solvent composition for extracting phenolic antioxidants from Cannabis sativa L. leaves and inflorescences. Three solvents (water, acetone, and ethanol) were systematically combined to evaluate how solvent polarity affects bioactive compound recovery, with extracts analyzed for total phenolic content (TPC) and antioxidant activity using five different assays (DPPH, ABTS, FRAP, CUPRAC, TAC), followed by chemical profiling using HPLC-DAD/ESI-MS2. **Key Findings** • Solvent composition significantly influenced extraction efficiency with organ-specific optimal mixtures: inflorescences achieved maximum TPC (16.05 mg GAE/g dry weight) with acetone-enriched solvent (2/3 acetone-1/6 water-1/6 ethanol), while leaves reached maximum TPC (13.65 mg GAE/g DW) with 2/3 acetone-1/3 ethanol. Multi-response optimization identified 1/3 water-1/3 ethanol-1/3 acetone for inflorescences and 50% water-30% ethanol-20% acetone for leaves as optimal formulations. • HPLC-DAD/ESI-MS2 analysis identified 32 bioactive compounds across multiple phytochemical classes including phenolic acids, flavonoids, hydroxycinnamic acid amides, lignanamides, and cannabinoid derivatives, with acetone-rich mixtures producing the highest total cannabinoid content (10.56 mg CBN equivalents/g DW for inflorescences and 8.45 mg CBN equivalents/g DW for leaves). • All extracts demonstrated notable antioxidant capacity across multiple assays (DPPH, ABTS, FRAP, CUPRAC, TAC), with activities correlating to phenolic content and showing clear organ and solvent-dependent selectivity. **Dosage & Administration** Not reported. **Safety & Adverse Effects** Not reported. **Evidence Quality** This is an in vitro extraction optimization study with limited clinical translational evidence. The research focuses on bench-top solvent chemistry and phytochemical characterization rather than human clinical outcomes. Strengths include comprehensive analytical methodology using multiple antioxidant assays and advanced mass spectrometry profiling. Limitations include lack of bioavailability data, absence of in vivo validation, and no assessment of safety or efficacy in human subjects. Results are specific to Moroccan cannabis cultivars and may not generalize across geographic origins or genetic varieties. The study provides valuable foundational data for standardizing extraction protocols but requires downstream pharmacological and clinical investigation to support pharmaceutical or nutraceutical applications.

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