contadores
Skip to main navigation menu Skip to main content Skip to site footer
##common.pageHeaderLogo.altText##

Research Article

Vol. 42 No. 1 (2025): Revista de Ciencias Agrícolas - Primer cuatrimestre, Enero - Abril 2025

Development and validation of a liquid chromatographic method to analyze cannabinoids in plants and oils

DOI
https://doi.org/10.22267/rcia.20254201.253
Submitted
July 27, 2022
Published
2025-04-28

Abstract

The therapeutic properties of Cannabis sativa have been widely investigated and have become a subject of great importance for the treatment of diseases such as epilepsy, as well as pain management in patients with cancer, arthritis, and Parkinson's disease, among others. The commercialization of Cannabis products has flourished due to these properties, highlighting the need to develop analytical methods for quality control of products marketed for medicinal purposes, as well as for Cannabis research. The objective of this work was to develop an HPLC-DAD-based analytical method for Cannabidiol (CBD), Cannabinol (CBN), and Tetrahydrocannabinol, (∆9-THC) cannabinoids tests in commercial Cannabis oils and plant material samples. Regarding the suitability of the system, the developed analytical method exhibited symmetrical peaks, with good resolution and symmetry. The system’s precision was adequate, with relative standard deviation (%RSD) values of less than 5%, and accuracy in terms of recovery greater than 90% for the extractions. Analysis of commercial samples revealed high ∆9-THC content (2.6-42.55 mg/mL) and low CBD content (0-4.3 mg/mL). The plant material sample analyzed correspond to a non-psychoactive chemotype (∆9-THC ≤ 1%) with a higher CBD content (8.0 mg/g). Considering the above, the developed method is suitable for the analysis of plant material and cannabis oils.

References

  1. Aizpurua-Olaizola, O.; Omar, J.; Navarro, P.; Olivares, M.; Etxebarria, N.; Usobiaga, A. (2014). Identification and quantification of cannabinoids in Cannabis sativa L. plants by high performance liquid chromatography-mass spectrometry. Analytical and bioanalytical chemistry. 406: 7549–7560. https://doi.org/10.1007/s00216-014-8177-x
  2. AOAC (2020). AOAC scientists approve Official Method of Analysis for cannabinoids in hemp. https://cutt.ly/fLupkdK
  3. Armentano P. (2015). Emerging Clinical Aplications for Cannabis and Canabinnoids. A Review of the Recent Scientific Literature. NORML Foundation. https://cutt.ly/rX7YwaT
  4. Attard, T.M.; Bainier, M.; Reinaud, A.; Lanot, S.J.; McQueen-Mason, A.; J. Hunt. (2018). Utilization of supercritical fluids for the effective extraction of waxes and Cannabidiol (CBD) from hemp wastes. Industrial Crops and Products. 112: 38–46. https://doi.org/10.1016/j.indcrop.2017.10.045
  5. Brighenti, V.; Pellati, F.; Steinbach,M.; Maran, D.; Benvenuti, S. (2017). Development of a new extraction technique and HPLC method for the analysis of non-psychoactive cannabinoids in fibre-type Cannabis sativa L. (hemp). Journal of Pharmaceutical and Biomedical Analysis. 143: 228–236. https://doi.org/10.1016/j.jpba.2017.05.049
  6. Bolchi,C.; Pallavicini, M.; Casagni, E.; De Manincor, E.; Gambaro, V.; Dei Cas, M.; Roda, G. (2021). Development and early identification of Cannabis chemotypes during the plant growth: Current Analytical and Chemometric Approaches. Analytical Sciences. 37:1665-1673. https://doi.org/10.2116/analsci.21R004
  7. Citti, C.; Linciano, P.; Panseri, S.; Vezzalini, F.; Forni, F.; Vandelli, M.A.; Cannazza G. (2019). Cannabinoid Profiling of Hemp Seed Oil by Liquid Chromatography Coupled to High-Resolution Mass Spectrometry. Frontiers in plant science. 10:120. https://doi.org/10.3389/fpls.2019.00120
  8. Florian, R.N.M; Parada, A.F.; Garzón, M.W. (2009). Study of cannabinoids content in marihuana samples (Cannabis sativa L.) cultivated in several regions of Colombia. Vitae. 16(2): 237-244. https://doi.org/10.17533/udea.vitae.1936
  9. Gallo-Molina, A.C.; Castro-Vargas, H.I.; Garzón-Méndez, W.F.; Martínez, J.A.; Rivera, Z.J.; King, J.W.; Parada-Alfonso, F. (2019). Extraction, isolation, and purification of tetrahydrocannabinol from the Cannabis sativa L. plant using supercritical fluid extraction and solid phase extraction. The Journal of Supercritical Fluids. 146: 208-216. https://doi.org/10.1016/j.supflu.2019.01.020
  10. González, S.; Sagredo, O.; Gómez, M.; Ramos, J.A. (2002). Química y metabolismo de los cannabinoides. In: Sociedad Española de Investigación sobre Cannabinoides (ed). Guía Básica sobre los Cannabinoides. Pp. 13-22. Universidad Complutense de Madrid. 160p Recuperada de: https://cutt.ly/GX7OSrC
  11. ICH Harmonised Tripartite Guideline (2005). International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use. Validation of Analytical Procedures: Text and Methodology Q2(R1).https://database.ich.org/sites/default/files/Q2%28R1%29%20Guideline.pdf
  12. Karanian, D.A.; Bahr, B.A. (2006). Cannabinoid drugs and enhancement of endocannabinoid responses: strategies for a wide array of disease states. Current Molecular Medicine. 6(6):84-677. https://doi.org/10.2174/156652406778194991
  13. Layton, C.; Reuter, W.M. (2017). Analysis of Cannabinoids in Hemp Seed Oils by HPLC using PDA Detection. Aplication note. https://acortar.link/N7EECy
  14. López, G.; Brindis, F.; Niizawa, C.; Ventura, R. (2014). Cannabis sativa L., una planta singular. Revista Mexicana de Ciencias Farmacéuticas. 45(4):1-6.
  15. Madej, K.; Kózka,G.; Winiarski, M.;Piekoszewski,W.(2020). A Simple, fast and green oil sample preparation method for determination of cannabidioloic acid and cannabidiol by HPLC-DAD. Separations. 7(4): 60. https://doi.org/10.3390/separations7040060
  16. Minsalud-Ministerio de Salud y Seguridad Social de Colombia. (2017). Decreto 613. Reglamentación para la fabricación, uso de semillas y cultivo de cannabis para fines medicinales y científicos.: https://cutt.ly/5X7SKFZ
  17. Niesink, R.J.; Van Laar, M.W. (2013). Does cannabidiol protect against adverse psychological effects of THC?. Frontiers in Psychiatry. 4: 130. https://doi.org/10.3389/fpsyt.2013.00130
  18. Olivas-Aguirre, M.; Gutiérrez-Iñiguez, C.; Pottosin, I.; Dobrovinskaya, 0. (2024). Molecular targets for cannabinoids in natural killer cells: Do they modulate the antitumor activity?. Receptors. 3(2): 122–144. https://doi.org/10.3390/receptors3020007
  19. Parmar, J.; Forrest, B.; Freeman, R. (2016). Medical marijuana patient counseling points for health care professionals based on trends in the medical uses, efficacy, and adverse effects of cannabis-based pharmaceutical drugs. Research in Social and Administrative Pharmacy. 12(4): 638-654 https://doi.org/10.1016/j.sapharm.2015.09.002
  20. Patel, B.; Wene, D.; Fan,Z.T. (2017). Qualitative and quantitative measurement of cannabinoids in cannabis using modified HPLC/DAD method. Journal of Pharmaceutical and Biomedical Analysis. 146: 15–23. https://doi.org/10.1016/j.jpba.2017.07.021
  21. Pavlovic, R.; Nenna, G.; Calvi, L.; Panseri, S.; Borgonovo, G.; Giupponi, L.; Cannazza, G.; Giorgi, A. (2018). Quality traits of “cannabidiol oils”: cannabinoids content, terpene fingerprint and oxidation stability of European commercially available preparations. Molecules. 23(5): 1230–51. https://doi.org/10.3390/molecules23051230
  22. Rubiano, D. R. (2019). Diseño de un plan de negocios para el cultivo, procesamiento y comercialización de cannabis medicinal. Fundación Universidad de América. https://cdn.technologynetworks.com/tn/Resources/pdf/cannabis-analysis-potency-testing-identification-and-quantification-of-thc-and-cbd-by-gcfid-and.pdf
  23. Ruppel, T.; Kuffel, N. (2016). Cannabis analysis: Potency testing identification and quantification of THC and CBD by GC/FID and GC/MS. Aplication note. https://acortar.link/72EypO
  24. Saingam, W.; Sakunpak, A. (2018). Development and validation of reverse phase high performance liquid chromatography method for the determination of delta-9-tetrahydrocannabinol and cannabidiol in oromucosal spray from cannabis extract. Revista Brasileira de Farmacognosia. 28(6): 669–672. https://doi.org/10.1016/j.bjp.2018.08.001
  25. Santos-Álvarez, I.; Pérez-Lloret, P.; González-Soriano, J.; Pérez-Moreno, M. (2021). Aproximación a la evaluación de la potencia de la resina de cannabis en Madrid: ¿un riesgo para la salud? Adicciones, 0. http://dx.doi.org/10.20882/adicciones.1630
  26. Teräsvalli H. (2020). Extraction and Purification of Cannabidiol. https://cutt.ly/PX7JD81
  27. UNODC. (2022). Recommended methods for the identification and analysis of cannabis and cannabis products. https://acortar.link/86rYtT
  28. Vaclavik, L.; Benes, F.; Fenclova, M.; Hricko, J.; Krmela, A.; Svobodova, V.; Hajslova, J.; Mastovska, K. (2019). Quantitation of cannabinoids in cannabis dried plant materials, concentrates, and oils using liquid chromatography-diode array detection technique with optional mass spectrometric detection: single-laboratory validation study, first action 2018.11. Journal of AOAC International. 102(6): 1822–1833. https://doi.org/10.5740/jaoacint.18-0426
  29. Velázquez, D.; Friman, N.; González, M. (2017). Polémica actual a nivel internacional sobre la legalización de la marihuana. Correo Científico Médico. 21(1): 269-272.
  30. Vučković, S.; Srebro, D.; Vujović, K.S.; Vučetić, Č.; Prostran, M. (2018). Cannabinoids and pain: New insights from old molecules. Frontiers in Pharmacology. 13(9): 1259. https://doi.org/10.3389/fphar.2018.01259
  31. Zivovinovic, S.; Alder, R.; Allenspach, M.D.; Steuer,C. (2018). Determination of cannabinoids in Cannabis sativa L. samples for recreational, medical, and forensic purposes by reversed-phase liquid chromatography-ultraviolet detection. Journal of Analytical Science and Technology. 9: 27. https://doi.org/10.1186/s40543-018-0159-8

Downloads

Download data is not yet available.