Acute oral toxicity of Cannabis sativa L. co-products in mice

Taha BALAFREJ; Souad SKALLI; Souad BENAICH; Asmae LABYAD; Rania AKOH; Fatima Zahra EL MSKINI; Slimane MEHDAD; Younes  ZAID; Oualid ABBOUSSI; Aziz MOUHADDACH; Saadia RAIS; Chaimae RAIS; Rachida HASSIKOU

doi:10.55779/nsb15311591

Authors

Taha BALAFREJ Mohammed V University in Rabat, Faculty of Sciences, Plant and Microbial Biotechnologies, Biodiversity, and Environment Center (MA)
Souad SKALLI Mohammed V University in Rabat, Faculty of Sciences, Plant and Microbial Biotechnologies, Biodiversity, and Environment Center (MA)
Souad BENAICH Mohammed V University in Rabat, Faculty of Sciences, Physiology and Physiopathology Team (MA)
Asmae LABYAD Mohammed V University in Rabat, Faculty of Sciences, Physiology and Physiopathology Team (MA)
Rania AKOH Mohammed V University in Rabat, Faculty of Sciences, Plant and Microbial Biotechnologies, Biodiversity, and Environment Center (MA)
Fatima Zahra EL MSKINI Mohammed V University in Rabat, Faculty of Sciences, Physiology and Physiopathology Team (MA)
Slimane MEHDAD Mohammed V University in Rabat, Faculty of Sciences, Physiology and Physiopathology Team (MA)
Younes ZAID Mohammed V University in Rabat, Faculty of Sciences, Plant and Microbial Biotechnologies, Biodiversity, and Environment Center (MA)
Oualid ABBOUSSI Mohammed V University in Rabat, Faculty of Sciences, Physiology and Physiopathology Team (MA)
Aziz MOUHADDACH Mohammed V University in Rabat, Faculty of Sciences, Plant and Microbial Biotechnologies, Biodiversity, and Environment Center (MA)
Saadia RAIS Sidi Mohamed Ben Abdellah University in Fes, Faculty of Sciences and Technologies, Laboratory of Functional Ecology and Environmental Engineering (MA)
Chaimae RAIS National Agency for Medicinal and Aromatic Plants, Laboratory of Botany (MA)
Rachida HASSIKOU Mohammed V University in Rabat, Faculty of Sciences, Plant and Microbial Biotechnologies, Biodiversity, and Environment Center (MA)

DOI:

https://doi.org/10.55779/nsb15311591

Keywords:

co-products, efficacy, extraction methods

Abstract

Cannabis sativa L. commonly known as ‘Kif’, is a medicinal plant belonging to the Cannabaceae family (Cannabis genus). It is used in traditional folk medicine to treat several diseases. The recent legalization of cannabis use for medicinal, cosmetic, and industrial purpose in Morocco (law 13.21), has enabled us to assess the potential acute toxicity of extract from its female plant co-products in Swiss mice. In this experimentation, 3 different extraction methods (maceration, decoction, and sonication) were used to determine the efficacy of cannabis extracts and investigate their acute toxicity in mice over a period of 14 days. Decoction showed the highest efficacy (19.7%), followed by maceration (16.2%), and finally sonication (5.4%). Regarding the toxicity results of the oral gavage administration dose at 2,000 mg kg^-1, there were no recorded deaths and no changes in the internal organs (kidneys and liver) between treated and control groups of both male and female mice. We also found that body weight of female mice treated by decoction and maceration extracts decreased significantly compared to those treated with sonication extract throughout the experimental period, when treated and untreated male mice show no significant changes (p-value < 0.05). Our findings suggest that the LD₅₀ may be greater than 2.000 mg kg^-1. Nevertheless, the extracts showed signs of low-level toxicity such as drowsiness, hypoactivity, and tachycardia. Further studies to determine the phytochemical composition of the three extracts of C. sativa and assess the chronic and sub-chronic toxicity should be carried out.

Metrics

Metrics Loading ...

References

Adesina I, Bhowmik A, Sharma H, Shahbazi A (2020). A review on the current state of knowledge of growing conditions, agronomic soil health practices and utilities of hemp in the United States. Agriculture 10(4):129. https://doi.org/10.3390/agriculture10040129.

Afsahi K (2017). La construction socio-économique du cannabis au Maroc: Le kif comme produit traditionnel, produit manufacturé et produit de contrebande [The socio-economic construction of cannabis in Morocco: Kif as a traditional product, manufactured product and contraband product]. Tempo Social 99‑114. https://doi.org/10.11606/0103-2070.ts.2017.125668.

Ahmed M, Ji M, Qin P, Gu Z, Liu Y, Sikandar A, Iqbal MF, Javeed A (2019). Phytochemical screening, total phenolic and flavonoids contents and antioxidant activities of Citrullus colocynthis L. and Cannabis sativa L. Applied Ecology & Environmental Research 17(3):6961-6979. http://dx.doi.org/10.15666/aeer/1703_69616979

Angelo A. Izzo, Francesca Borrelli, Raffaele Capasso, Vincenzo Di Marzo, Raphael Mechoulam (2009). Non-psychotropic plant cannabinoids: new therapeutic opportunities from an ancient herb. Trends in Pharmacological Sciences 30(10):515-527. https://doi.org/10.1016/j.tips.2009.07.006.

Babiker EE, Uslu N, Al Juhaimi F, Ahmed IA, Ghafoor K, Özcan MM, Almusallam IA (2021). Effect of roasting on antioxidative properties, polyphenol profile and fatty acids composition of hemp (Cannabis sativa L.) seeds. LWT-Food Science and Technology 139:110537. https://doi.org/10.1016/j.lwt.2020.110537.

Bailey MM, Emily Mills MC, Haas AE, Bailey K, Kaufmann RC (2022). The effects of subacute exposure to a water-soluble cannabinol compound in male mice. Journal of Cannabis Research 4(1):44. https://doi.org/10.1186/s42238-022-00153-w.

Bailoni L, Bacchin E, Trocino A, Arango S (2021). Hemp (Cannabis sativa L.) seed and co-products inclusion in diets for dairy ruminants: A review. Animals 11(3):856. https://doi.org/10.3390/ani11030856.

Bellakhdar Jamal (1997). Contribution à l’étude de la pharmacopée traditionnelle au Maroc : la situation actuelle, les produits, les sources du savoir (enquête ethnopharmacologique de terrain réalisée de 1969 à 1992) [Contribution to the study of traditional pharmacopoeia in Morocco: the current situation, products, sources of knowledge (ethnopharmacological field survey carried out from 1969 to 1992)]. PhD thesis, Paul Verlaine university, Metz.

Brunetti P, Pichini S, Pacifici R, Busardò FP, Del Rio A (2020). Herbal preparations of medical cannabis: a vademecum for prescribing doctors. Medicina 56(5):237. https://doi.org/10.3390/medicina56050237

Cavalli J, Dutra RC (2021). A closer look at cannabimimetic terpenes, polyphenols, and flavonoids: a promising road forward. Neural Regeneration Research 16(7):1433-1435. https://doi.org/10.4103/1673-5374.301011

Centre Anti Poison et de Pharmacovigilance du Maroc (2015). Rapport annuel des analyses toxicologiques de l’année 2015 [Annual report of toxicological analyses of the year 2015]. Toxicologie Maroc. Retrieved 2023 May 24 from: https://www.capm-sante.ma/uploads/documents/34.pdf

Crini G, Lichtfouse E, Chanet G, Morin-Crini N (2020). Applications of hemp in textiles, paper industry, insulation and building materials, horticulture, animal nutrition, food and beverages, nutraceuticals, cosmetics and hygiene, medicine, agrochemistry, energy production and environment: A review. Environmental Chemistry Letters 18(5):1451-1476. https://doi.org/10.1007/s10311-020-01029-2

Crocq MA (2022). 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.

EMCDDA (2019). Usage médical du cannabis et des cannabinoïdes : questions et réponses à l’intention des décideurs politiques [Medical use of cannabis and cannabinoids: questions and answers for policymakers]. Luxembourg Publications Office of the European Union. Retrieved 2023 May 25 from: https://data.europa.eu/doi/10.2810/988921.

Pellati F, Borgonetti V, Brighenti V, Biagi M, Benvenuti S, Corsi L (2018). Cannabis sativa L. and nonpsychoactive cannabinoids: their chemistry and role against oxidative stress, inflammation, and cancer. BioMed Research International 2018:1691428. https://doi.org/10.1155/2018/1691428.

Gamelin F-X, Berthoin S, Cuvelier G, Mendes A, Anthierens A, Heyman E (2021). L’usage du cannabidiol dans le sport: Une bonne idée? [The use of cannabidiol in sport: A good idea?]. 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.

Hall W (2015). What has research over the past two decades revealed about the adverse health effects of recreational cannabis use? Addiction 110(1):19-35. https://doi.org/10.1111/add.12703

Hassikou R, Oulladi H, Arahou M (2014). Activité antimycosique des extraits du chêne-liège Quercus suber sur Trichophyton rubrum et Candida albicans. Phytothérapie 12(4):206 212. https://doi.org/10.1007/s10298-014-0874-2.

Hodge A, Sterner B (2005). Toxicity classes. In: Canadian center for occupational Health and safety. Retrieved 2023 July 31 from: http://www.ccohs.ca/oshanswers/chemicals/id50.htm

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

Huestis MA, Solimini R, Pichini S, Pacifici R, Carlier J, Busardò FP (2019). Cannabidiol adverse effects and toxicity. Current Neuropharmacology 17(10):974-989. https://doi.org/10.2174/1570159X17666190603171901

Jin D, Dai K, Xie Z, Chen J (2020). Secondary metabolites profiled in cannabis inflorescences, leaves, stem barks, and roots for medicinal purposes. Scientific Reports 10(1):3309. https://doi.org/10.1038/s41598-020-60172-6.

Kifayatullah M, Mustafa MS, Sengupta P, Sarker MMR, Das A, Das SK (2015). Evaluation of the acute and sub-acute toxicity of the ethanolic extract of Pericampylus glaucus (Lam.) Merr. in BALB/c mice. Journal of Acute Disease 4(4):309-315. https://doi.org/10.1016/j.joad.2015.06.010.

Liktor-Busa E, Keresztes A, LaVigne J, Streicher JM, Largent-Milnes TM (2021). Analgesic potential of terpenes derived from Cannabis sativa. Pharmacological Reviews 73(4):1269-1297. https://doi.org/10.1124/pharmrev.120.000046.

Manthey J, Freeman TP, Kilian C, López-Pelayo H, Rehm J (2021). Public health monitoring of cannabis use in Europe: prevalence of use, cannabis potency, and treatment rates. The Lancet Regional Health-Europe 10:100227. https://doi.org/10.1016/j.lanepe.2021.100227

Maqbool J, Anwar H, Rasul A, Imran A, Saadullah M, Malik SA, … Islam S (2023). Comparative evaluation of ethyl acetate and n-Hexane extracts of Cannabis sativa L. leaves for muscle function restoration after peripheral nerve lesion. Food Science and Nutrition. https://doi.org/10.1002/fsn3.3255.

Merzouki A, Ed-Derfoufi F, Mesa JM (2000). Hemp (Cannabis sativa L.) and abortion. Journal of Ethnopharmacology 73(3):501-503. https://doi.org/10.1016/S0378-8741(00)00323-8

Noble MJ, Hedberg K, Hendrickson RG (2019). Acute cannabis toxicity. Clinical Toxicology 57(8):735-742. https://doi.org/10.1080/15563650.2018.1548708.

OCDE (2022). Lignes directrices de l'OCDE pour les essais de produits chimiques, Section 4: Effets sur la santé. Essai n° 425 : Toxicité aiguë par voie orale : méthode de l’ajustement des doses [OECD guidelines for the testing of chemicals, section 4: Health effects. Test no. 425: Acute oral toxicity: dose adjustment method]. Retrieved 2023 May 24 from: https://read.oecd-ilibrary.org/environment/essai-n-425-toxicite-aigue-par-voie-orale-methode-de-l-ajustement-des-doses_9789264071056-fr

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.

Ouanouche EH, Elmostafi H, Amarat N, Wafaa O, Ryad T, El Hessni A, Mesfioui A (2022). Cannabis and schizophrenia: characterisation of a risk factor in a sample of Moroccan patients hospitalized for psychosis. Middle East Current Psychiatry 29(1):1-8. https://doi.org/10.1186/s43045-022-00173-5.

Rehman M, Fahad S, Du G, Cheng X, Yang Y, Tang K, … Deng G (2021). Evaluation of hemp (Cannabis sativa L.) as an industrial crop: A review. Environmental Science and Pollution Research 28(38):52832-43. https://doi.org/10.1007/s11356-021-16264-5

Rožanc J, Kotnik P, Milojević M, Gradišnik L, Knez Hrnčič M, Knez Ž, Maver U (2021). Different Cannabis sativa extraction methods result in different biological activities against a colon cancer cell line and healthy colon cells. Plants 10(3):566. https://doi.org/10.3390/plants10030566.

Saleem U, Amin S, Ahmad B, Azeem H, Anwar F, Mary S (2017). Acute oral toxicity evaluation of aqueous ethanolic extract of Saccharum munja Roxb. Roots in albino mice as per OECD 425 TG. Toxicology Reports 4:580‑585. https://doi.org/10.1016/j.toxrep.2017.10.005

Sazmand M, Mehrabani D, Hosseini E, Amini M (2018). The effect of hydroalcoholic extract of Cannabis sativa on morphology and growth of bone marrow mesenchymal stem cells in rat. Electronic Journal of General Medicine 15(3). https://doi.org/10.29333/ejgm/86195

Schep LJ, Slaughter RJ, Glue P, Gee P (2020). The clinical toxicology of cannabis. The New Zealand Medical Journal (Online) 133(1523):96-103.

Thompson GR, Rosenkrantz H, Schaeppi UH, Braude MC (1973). Comparison of acute oral toxicity of cannabinoids in rats, dogs and monkeys. Toxicology and Applied Pharmacology 25(3):363‑372. https://doi.org/10.1016/0041-008X(73)90310-4.

UNODC (2022). Recommended methods for the identification and analysis of cannabis and cannabis products. Vienna: United nations office on drugs and crime. Retrieved 2023 may 20 from: https://www.unodc.org/unodc/en/scientist/recommended-methods-for-the-identification-and-analysis-of-cannabis-and-cannabis-products.html.

Valizadehderakhshan M, Shahbazi A, Kazem-Rostami M, Todd MS, Bhowmik A, Wang L (2021). Extraction of cannabinoids from Cannabis sativa L. (Hemp). Agriculture 11(5):384. https://doi.org/10.3390/agriculture11050384.

Vastolo A, Calabrò S, Pacifico S, Koura BI, Cutrignelli MI (2021). Chemical and nutritional characteristics of Cannabis sativa L. co‐products. Journal of Animal Physiology and Animal Nutrition 105:1-9. https://doi.org/10.1111/jpn.13557.

Visković J, Zheljazkov VD, Sikora V, Noller J, Latković D, Ocamb CM, Koren A (2023). Industrial hemp (Cannabis sativa L.) agronomy and utilization: A review. Agronomy 13(3):931. https://doi.org/10.3390/agronomy13030931.

Wang GS, Banerji S, Contreras AE, Hall KE (2020). Marijuana exposures in Colorado, reported to regional poison centre, 2000–2018. Injury Prevention 26(2):184-186. https://doi.org/10.1136/injuryprev-2019-043360.

World Health Organization (2016). The health and social effects of nonmedical cannabis use. Retrieved 2023 July 31 from: https://apps.who.int/iris/bitstream/handle/10665/251056/9789241510240-eng.pdf?sequence=1&isAllowed=y

World health organization et WHO expert committee on drug dependence (2018). WHO expert committee on drug dependence: fortieth report. WHO Technical Report Series: 1013. Geneva: Health Organization. Retrieved 2023 May 20 from: https://apps.who.int/iris/handle/10665/279948.

Yassa HA, Dawood AEWA, Shehata MM, Abdel-Hady RH, Abdel Aal KM (2010). Subchronic toxicity of cannabis leaves on male albino rats. Human & Experimental Toxicology 29(1):37-47. https://doi.org/10.1177/0960327109354312.

Zimmermann M (1983). Ethical guidelines for investigations of experimental pain in conscious animals. Pain 16(2):109‑110. https://doi.org/10.1016/0304-3959(83)90201-4.