Abstract
Most of the pharmacological effects of Cannabis species have been attributed to their cannabinoid constituents, primarily Δ9-tetrahydrocannabinol and cannabidiol. Efficient methods of extraction, purification, and characterization of these compounds are necessary to obtain both enriched preparations and isolated compounds for therapeutic applications. In this study, forensic samples of Cannabis sativa L., Cannabaceae, were subjected to extraction, using maceration in two different solvents, methanol, and diethyl ether. These extracts were subjected to thermal decarboxylation and purification by liquid–liquid fractionation. Δ9-Tetrahydrocannabinol and cannabidiol concentrations from various extracts were quantified by high-performance liquid chromatography with UV detection. Extract cytotoxicity was assayed in vitro using human erythrocytes, mouse embryonic fibroblasts (NIH3T3), and murine melanoma (B16F10) cells. The B16F10 cell death mechanism was also evaluated after treatment with extract samples. The results demonstrated promising effects of various cannabinoid extracts in a murine melanoma cancer line.
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Adapted from De Backer et al. (2009)
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Acknowledgements
Analyses of DNA extraction, amplification, and fragment were performed in the Human and Molecular Genetics Laboratory of Pontifícia Universidade Católica do Rio Grande do Sul by Lucas de O. P. Ribeiro and Clarice Sampaio Alho.
Funding
Financial support was provided by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Conselho Nacional de Pesquisa e Desenvolvimento (CNPq), and Fundação de Amparo à Pesquisa e Inovação do Estado de Santa Catarina (FAPESC), Brazil.
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PRM, FRMS, and PAA: development/execution of methodology for Cannabis extraction; FRMS: HPLC analysis; RS: development and verification, of the overall replication/reproducibility of results by HPLC analysis; MEM, FKR, and PEF: evaluation/execution of in vitro tests; ABM: conceptualization regarding the use of seized Cannabis as a source of raw material for extract preparation; RAM-de-Á: reagent supply, laboratory materials, instrumentation, and other analytical tools for in vitro analysis; JWB: manuscript preparation, critical review, commentary, and revision; PEF and FKR: application of statistical, mathematical, computational, or other formal techniques to analyze or synthesize study data; FKR and PAA: conceptualization and evolution of overarching research goals and aims; PAA: management and coordination of the research project.
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All studies in this manuscript were in accordance with the Institutional Review Board and Institutional Animal Care and Use Committee procedures. Experiments reported in this study were approved by the medical ethics committee of UNESC (Criciúma, Brazil), protocol number CAAE 14560419.9.0000.0119.
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Milanez, P.R., da Silva, F.M.R., Scussel, R. et al. Cannabis Extracts and Their Cytotoxic Effects on Human Erythrocytes, Fibroblasts, and Murine Melanoma. Rev. Bras. Farmacogn. 31, 750–761 (2021). https://doi.org/10.1007/s43450-021-00200-2
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DOI: https://doi.org/10.1007/s43450-021-00200-2