Moroccan Cannabis sativa L. seed chloroform extract: phytochemistry, acute toxicity, and metabolic-behavioral effects in mice
DOI:
https://doi.org/10.55779/nsb18212928Keywords:
acute toxicity, cannabidiol, Cannabis sativa L., hydrocarbons, phytocannabinoids, seedsAbstract
Despite growing interest in the therapeutic use of Cannabis sativa L. following its legalization for medicinal purposes in 2021, the toxicity profile of its seeds remains largely underexplored in Morocco. This study aimed to characterize the phytochemical profile and acute oral toxicity of a chloroform seed extract (C/S) from Moroccan Cannabis sativa L. C/S extract (yield: 4.27 ± 0.13%) was analyzed by gas chromatography coupled with mass spectrometry (GC-MS). Acute toxicity testing was performed according to OECD Test Guideline 425: female Swiss mice (n = 5/group) received a single 2,000 mg kg⁻¹ oral dose or vehicle control, followed by 14 d monitoring of clinical signs, body weight (BW), organ weight, and serum biochemistry (ASAT/ALAT, creatinine, urea, and lipid profile). GC-MS analysis revealed 16 compounds, with octacosane (6.80%), 2-methyloctadecane (5.75%), and cannabidiol (5.32%) as the major components. No treatment-related mortality was observed (LD₅₀ > 2,000 mg kg⁻¹), and only transient signs of hypoactivity/tachycardia were noted, which resolved spontaneously within a few hours. BW gain (p = 0.154), liver/BW (p = 0.103), and kidney/BW (p = 0.406) ratios showed no significant differences. Total cholesterol was lower in the C/S group than in controls (0.82 ± 0.06 g L⁻¹ vs. 2.13 ± 0.38 g L⁻¹, p = 0.014), whereas LDL-C was higher in the C/S group (0.07 ± 0.00 g L⁻¹ vs. 0.03 ± 0.01 g L⁻¹, p = 0.002). The C/S extract demonstrated low acute toxicity under the tested conditions, supporting further safety-oriented pharmaceutical investigation of Moroccan Cannabis sativa L. seeds.
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References
Akinlade OM, Owoyele BV, Soladoye AO (2021). Streptozotocin-induced type 1 and 2 diabetes in rodents: a model for studying diabetic cardiac autonomic neuropathy. African Health Sciences, 21(2): 719-727. https://doi.org/10.4314/ahs.v21i2.30
Bachir F, Eddouks M, Arahou M, Fekhaoui M (2022). Origin, early history, cultivation, and characteristics of the traditional varieties of Moroccan Cannabis sativa L. Cannabis and Cannabinoid Research, 7(5): 603-615. https://doi.org/10.1089/can.2021.0020
Balachandran A, Choi SB, Beata MM, Małgorzata J, Froemming GRA, Lavilla CA, et al. (2023). Antioxidant, wound healing potential and in silico assessment of naringin, eicosane and octacosane. Molecules, 28(3): 1043. https://doi.org/10.3390/molecules28031043
Balafrej T, Skalli S, Benaich S, Labyad A, Akoh R, El Mskini FZ, et al. (2023). Acute oral toxicity of Cannabis sativa L. co-products in mice. Notulae Scientia Biologicae, 15(3): 11591. https://doi.org/10.55779/nsb15311591
Burstein S (2015). Cannabidiol (CBD) and its analogs: a review of their effects on inflammation. Bioorganic & Medicinal Chemistry, 23(7): 1377-1385. https://doi.org/10.1016/j.bmc.2015.01.059
Chen S, Kim JK (2024). The role of cannabidiol in liver disease: a systemic review. International Journal of Molecular Sciences, 25(4): 2370. https://doi.org/10.3390/ijms25042370
Crocq MA (2020). History of cannabis and the endocannabinoid system. Dialogues in Clinical Neuroscience, 22(3): 223-228. https://doi.org/10.31887/DCNS.2020.22.3/mcrocq
Ebrahimi N, Far NP, Fakhr SS, Faghihkhorasani F, Miraghel SA, Chaleshtori SR, et al. (2023). The endocannabinoid system, a new gatekeeper in the pharmacology of human hepatocellular carcinoma. Environmental Research, 228: 115914. https://doi.org/10.1016/j.envres.2023.115914
El-Mernissi R, El Menyiy N, Zouhri A, El-Mernissi Y, Metouekel A, Siddique F, et al. (2026). A comparative study on phytochemical analysis and biological properties of three varieties of Cannabis sativa L. seeds. Open Life Sciences, 21(1): 20251211. https://doi.org/10.1515/biol-2025-1211
El-Mernissi R, El Menyiy N, Moubachir N, Zouhri A, El-Mernissi Y, Siddique F, et al. (2024). Cannabis sativa L. essential oil: chemical composition, antioxidant, antimicrobial properties, and acute toxicity: in vitro, in vivo, and in silico study. Open Chemistry, 22(1): 20230214. https://doi.org/10.1515/chem-2023-0214
El-Mernissi R, El Menyiy N, Zouhri A, El-Mernissi Y, Diai F, Siddique F, et al. (2024). Phytochemical profiling and bioactivity evaluation of CBD- and THC-enriched Cannabis sativa extracts: in vitro and in silico investigation of antioxidant and anti-inflammatory effects. Open Chemistry, 22(1): 20240119. https://doi.org/10.1515/chem-2024-0119
Ewing LE, Skinner CM, Quick CM, Kennon-McGill S, McGill MR, Walker LA, et al. (2019). Hepatotoxicity of a cannabidiol-rich Cannabis extract in the mouse model. Molecules, 24(9): 1694. https://doi.org/10.3390/molecules24091694
Gamelin FX, Berthoin S, Cuvelier G, Mendes A, Anthierens A, Heyman E (2021). L'usage du cannabidiol dans le sport: une bonne idée?. Science & Sports, 36(4): 251-258. https://doi.org/10.1016/j.scispo.2021.03.003
Gugliandolo A, Pollastro F, Grassi G, Bramanti P, Mazzon E (2018). In vitro model of neuroinflammation: efficacy of cannabigerol, a non-psychoactive cannabinoid. International Journal of Molecular Sciences, 19(7): 1992. https://doi.org/10.3390/ijms19071992
Hasan MM, Tama RT, Dona HA, Hoque NS, Rahaman MA, Alam MA (2025). Comprehensive review of phthalate exposure: health implications, biomarker detection and regulatory standards. The Journal of Steroid Biochemistry and Molecular Biology, 247: 106671. https://doi.org/10.1016/j.jsbmb.2024.106671
Hourfane S, Mechqoq H, Bekkali AY, Rocha JM, El Aouad N (2023). A comprehensive review on Cannabis sativa ethnobotany, phytochemistry, molecular docking and biological activities. Plants, 12(6): 1245. https://doi.org/10.3390/plants12061245
Huang ZR, Lin YK, Fang JY (2009). Biological and pharmacological activities of squalene and related compounds: potential uses in cosmetic dermatology. Molecules, 14(1): 540-554. https://doi.org/10.3390/molecules14010540
Ibork H, Ait Lhaj Z, Siddique F, El Idrissi S, Khallouki F, El Mernissi R, et al. (2025). Analgesic and toxicological evaluation of cannabidiol-rich Moroccan Cannabis sativa L. (Khardala variety) extract: evidence from an in vivo and in silico study. Open Life Sciences, 20(1): 20251141. https://doi.org/10.1515/biol-2025-1141
Jiao S, Cole TG, Kitchens RT, Pfleger B, Schonfeld G (1990). Genetic heterogeneity of lipoproteins in inbred strains of mice: analysis by gel-permeation chromatography. Metabolism: Clinical and Experimental, 39(2): 155-160. https://doi.org/10.1016/0026-0495(90)90069-o
Khalil A, Al Toufaily S, Shebaby W, El Hage M, Mroue D, Faour W, Mroueh M (2025). Lebanese Cannabis sativa L. extract protects from cisplatin-induced nephrotoxicity in mice by inhibiting podocyte apoptosis. Journal of Cannabis Research, 7(1): 3. https://doi.org/10.1186/s42238-025-00260-4
LaVigne JE, Hecksel R, Keresztes A, Streicher JM (2021). Cannabis sativa terpenes are cannabimimetic and selectively enhance cannabinoid activity. Scientific Reports, 11(1): 8232. https://doi.org/10.1038/s41598-021-87740-8
Lima KSB, Silva MEGDC, Araújo TCDL, Silva CPDF, Santos BL, Ribeiro LADA, et al. (2021). Cannabis roots: pharmacological and toxicological studies in mice. Journal of Ethnopharmacology, 271: 113868. https://doi.org/10.1016/j.jep.2021.113868
Majewski M, Jurgoński A (2021). The effect of hemp (Cannabis sativa L.) seeds and hemp seed oil on vascular dysfunction in obese male Zucker rats. Nutrients, 13(8): 2575. https://doi.org/10.3390/nu13082575
Musetti B, Kun A, Menchaca D, Rodríguez-Haralambides A, Varela J, Thomson L, Bahnson EM (2024). Cannabis sativa extracts inhibit LDL oxidation and the formation of foam cells in vitro, acting as potential multi-step inhibitors of atherosclerosis development. PLoS ONE, 19(12): e0310777. https://doi.org/10.1371/journal.pone.0310777
Odieka AE, Obuzor GU, Oyedeji OO, Gondwe M, Hosu YS, Oyedeji AO (2022). The medicinal natural products of Cannabis sativa Linn.: a review. Molecules, 27(5): 1689. https://doi.org/10.3390/molecules27051689
OECD (2022). Test No. 425: acute oral toxicity: up-and-down procedure. OECD Guidelines for the Testing of Chemicals, Section 4, OECD Publishing, Paris. Available at: https://www.oecd-ilibrary.org/environment/test-no-425-acute-oral-toxicity-up-and-down-procedure_9789264071049-en (Accessed: 18 October 2023).
Okulmus C, Icil NI, Turkyilmaz O, Yildirir ZT (2025). Clinical chemistry reference intervals for Swiss albino strain mice commonly used in scientific studies. Comparative Clinical Pathology, 34(4): 567-574. https://doi.org/10.1007/s00580-025-03683-w
Osei-Hyiaman D, DePetrillo M, Pacher P, Liu J, Radaeva S, Bátkai S, et al. (2005). Endocannabinoid activation at hepatic CB1 receptors stimulates fatty acid synthesis and contributes to diet-induced obesity. The Journal of Clinical Investigation, 115(5): 1298-1305. https://doi.org/10.1172/JCI23057
Pan H, Mukhopadhyay P, Rajesh M, Patel V, Mukhopadhyay B, Gao B, et al. (2009). Cannabidiol attenuates cisplatin-induced nephrotoxicity by decreasing oxidative/nitrosative stress, inflammation, and cell death. The Journal of Pharmacology and Experimental Therapeutics, 328(3): 708-714. https://doi.org/10.1124/jpet.108.147181
Panee J, Pomozi V, Franke AA, Le Saux O, Gerschenson M (2020). Chronic marijuana use moderates the correlations of serum cholesterol with systemic mitochondrial function and fluid cognition. Mitochondrion, 52: 135-143. https://doi.org/10.1016/j.mito.2020.03.006
Papada E (2025). The effects of terpenes on metabolism: a comprehensive review on recent updates. Current Opinion in Clinical Nutrition and Metabolic Care, 28(4): 323-329. https://doi.org/10.1097/MCO.0000000000001129
Poyatos L, Pérez-Acevedo AP, Papaseit E, Pérez-Mañá C, Martin S, Hladun O, et al. (2020). Oral administration of cannabis and Δ-9-tetrahydrocannabinol (THC) preparations: a systematic review. Medicina, 56(6): 309. https://doi.org/10.3390/medicina56060309
Raoui K, Kabdy H, Ettitaou A, Aitbaba A, Baslam A, Benrazzouk K, et al. (2024). Assessment of Moroccan Cannabis sativa seed oil: chemical analysis and evaluation of antioxidant, toxicological, and antinociceptive effects. Chemistry & Biodiversity, 21(8): e202400591. https://doi.org/10.1002/cbdv.202400591
Rosso E, Armone R, Costale A, Meineri G, Chiofalo B (2024). Hemp seed (Cannabis sativa L.) varieties: lipids profile and antioxidant capacity for monogastric nutrition. Animals, 14(18): 2699. https://doi.org/10.3390/ani14182699
Russo EB (2011). Taming THC: potential cannabis synergy and phytocannabinoid-terpenoid entourage effects. British Journal of Pharmacology, 163(7): 1344-1364. https://doi.org/10.1111/j.1476-5381.2011.01238.x
Sañudo-Peña MC, Romero J, Seale GE, Fernandez-Ruiz JJ, Walker JM (2000). Activational role of cannabinoids on movement. European Journal of Pharmacology, 391(3): 269-274. https://doi.org/10.1016/S0014-2999(00)00044-3
Simopoulos AP (2008). The importance of the omega-6/omega-3 fatty acid ratio in cardiovascular disease and other chronic diseases. Experimental Biology and Medicine, 233(6): 674-688. https://doi.org/10.3181/0711-MR-311
Sirangelo TM, Diretto G, Fiore A, Felletti S, Chenet T, Catani M, et al. (2025). Nutrients and bioactive compounds from Cannabis sativa seeds: a review focused on omics-based investigations. International Journal of Molecular Sciences, 26(11): 5219. https://doi.org/10.3390/ijms26115219
Soliman NA, El Dahmy SI, Shalaby AA, Mohammed KA (2024). Prospective affirmative therapeutics of cannabidiol oil mitigates doxorubicin-induced abnormalities in kidney function, inflammation, and renal tissue changes. Naunyn-Schmiedeberg's Archives of Pharmacology, 397(6): 3897-3906. https://doi.org/10.1007/s00210-023-02836-4
Tănase Apetroaei V, Pricop EM, Istrati DI, Vizireanu C (2024). Hemp seeds (Cannabis sativa L.) as a valuable source of natural ingredients for functional foods - a review. Molecules, 29(9): 2097. https://doi.org/10.3390/molecules29092097
Visković J, Zheljazkov VD, Sikora V, Noller J, Latković D, Ocamb CM, Koren M (2023). Industrial hemp (Cannabis sativa L.) agronomy and utilization: a review. Agronomy, 13(3): 931. https://doi.org/10.3390/agronomy13030931
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Copyright (c) 2026 Fatima Zahra El Mskini, Anass Rami, Asmae Labyad, Hind Ibork, Zakaria Ait Lhaj, Ahmed Oubaasri, Abdelmounaim Laabar, Aichetou Bouh, Nouriya El Ghoulam, Rachida Hassikou, Slimane Mehdad, Oualid Abboussi, Souad Benaich
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