Terpenoid Profiling of Thai Strain Cannabis Leaves (Cannabis sativa L. subsp. sativa) by Headspace (HS) Couple with GC/MS

Phurin  Watanakul; Jessada  Phattaralerphong; Kitsada  Pitija; Pattarawadee Sumthong Nakmee

doi:10.31436/jop.v4i1.225

Authors

Phurin Watanakul Faculty of Science at Sriracha, Kasetsart University Sriracha Campus, 199 Moo 6, Sukhumvit Rd., Sriracha, Chonburi, 20230, Thailand.
Jessada Phattaralerphong Faculty of Natural Resource and Agricultural Industry, Kasetsart University, Chalermprakiat Campus, Chiang Khruea, Muang, Sakon Nakhon, 47000, Thailand.
Kitsada Pitija PerkinElmer Scientific (Thailand) Co., Ltd. (Head Office) No. 290 Soi Soonvijai 4, Bangkapi, Huay Kwang, Bangkok, 10310, Thailand.
Pattarawadee Sumthong Nakmee Faculty of Science at Sriracha, Kasetsart University Sriracha Campus, 199 Moo 6, Sukhumvit Rd., Sriracha, Chonburi, 20230, Thailand. https://orcid.org/0000-0003-0315-2512

DOI:

https://doi.org/10.31436/jop.v4i1.225

Keywords:

Cannabis sativa, terpenoids, Headspace-gas chromatography/mass spectrometry (HS-GC/MS)

Abstract

Introduction: Cannabis terpenoids, especially volatile terpenes, were used for the classification of cannabis strains. The leaves of Cannabis sativa L. subsp. sativa Thai strain ‘Hang Krarok’ are used legally in traditional Thai medicines, cosmetics, and food ingredients in Thailand under the control of the tetrahydrocannabinol (if lower than 0.2% dry weight). One of the specific characteristics of this plant is the volatile oil which consists of mono-and the sesqui-terpenoids.

Materials and methods: Fresh cannabis leaves were ground and 1 g samples were kept in gas chromatography/mass spectrometry glass vials at 4 °C prior to measurement using headspace.

Results: More than 50 terpenoids were identified from the fresh leaves in the cannabis samples. The major compounds were ?–ocimene, L–limonene, terpinolene, p–cymenene, ?–(E)–caryophyllene, (Z,E)–?–farnesene, ?–bisabolene, and (E)–?–bisabolene.

Conclusion: The variation in the unique terpenoids in the Thai strain could be used in novel medicines and food and cosmetic products.

References

Adams, R. P. (2001). “Identification of essential oil components by gas chromatography/quadrupole mass spectrometry,” 4th Edition, Allured, Allured Publishing Corporation, Illinoi, USA, 804 p.

Andreani, S., Cian, M.C.D., Paolini, J., Desjobert, J.M., Costa, J., & Muselli, A. (2013). Chemical Variability and Antioxidant Activity of Limbarda crithmoides L. Essential Oil from Corsica. Chem. Biodivers. 10(11), 2061–2077. https://doi.org/10.1002/cbdv.201300109

Booth, J.K., & Bohlmann, J. (2019). Terpene in Cannabis sativa ?from plant genome to humans. Plant Sci. 284, 67–72. https://doi.org/10.1016/j.plantsci.2019.03.022

Booth, J.K., Page, J.E., & Bohlmann, J. (2017). Terpene synthases from Cannabis sativa. PLoS ONE 12(3), e0173911. https://doi.org/10.1371/journal.pone.0173911

Borges, C.V., Minatel, I.O., Gomez-Gomez, H.A., & Lima, G.P.P. (2017). Medicinal Plants: Influence of Environmental Factors on the Content of Secondary Metabolites. In Ghorbanpour, M., Varma, A. (eds). Medicinal Plants and Environmental Challenges. Springer, AG. pp 259–277. https://doi.org/10.1007/978-3-319-68717-9_15

Brousseau, V.D., Wu, B.S. Pherson, S.M., Morello, V., & Lefsrud, M. (2021). Cannabinoids and Terpenes: How Production of Photo-Protectants can be Manipulated to Enhance Cannabis sativa L. Phytochemistry. Front. Plant Sci. 12, 620021. https://doi.org/10.3389/fpls.2021.620021

Cassano, S., & Grassi, G. (2011). Variations in Terpene Profiles of Different Strains of Cannabis sativa L. Acta Horticulturae 925, 115–122. https://doi:10.17660/ActaHortic.2011.925.15

Duangdamrong, J, Jamparngernthaweesri, K, Sathiyos, Y., Poopong, S., Lumlerdkij, N., & Akarasereenont, P. (2022). Evidence of Cannabis for Medical Uses and Its Indications: An Evidence Based Comparison Between Thai Traditional Medicine and Western Medicine. Siriraj Medical Bulletin. 15(3), 155–163. https://doi.org/10.33192/smb.v15i3.256111

Dziok, M.Z., Bujak, T., Ziemlewska, A., & ?ukaszewska, Z.N. (2021). Positive Effect of Cannabis sativa L. Herb Extracts on Skin Cells and Assessment of Cannabinoid-Based Hydrogels Properties. Molecules. 26(4) 802. https://doi.org/10.3390/molecules26040802

Ehrensing, D. T. (1998). Feasibility of Industrial Hemp Production in the United States Pacific Northwest. In Station Bulletin 681. Agricultural Experiment Station, Oregon State University, OR, USA, p 1–41.

Fischedick, J.T. (2017). Identification of Terpenoid Chemotypes Among High (?) ? trans?D9?Tetrahydrocannabinol?Producing Cannabis sativa L. Cultivars. Cannabis Cannabinoid Res. 2(1), 34?47. https://doi: 10.1089/can.2016.0040

Fischedick, J.T., Hazekamp, A., Erkelens, T., Choi, Y.H., & Verpoorte, R. (2010). Metabolic fingerprinting of Cannabis sativa L., cannabinoids and terpenoids for chemotaxonomic and drug standardization purposes. Phytochemistry 71, 2058–2073. https://doi:10.1016/j.phytochem.2010.10.001

Gaggiotti, S., Palmieri, S., Pelle, F.D., Sergi, M., Cichelli, A., Mascini, M., & Compagnone, D. (2020). Piezoelectric peptide?hpDNA based electronic nose for the detection of terpenes; Evaluation of the aroma profile in different Cannabis sativa L.(hemp) samples. 308, 127697. https://doi.org/10.1016/j.snb.2020.127697

Gallily, R., Yetkhtin, Z., & Hanus O.L. (2018). The Anti–Inflammatory Properties of Terpenoids from Cannabis. Cannabis Cannabinoid Res. 3(1), 282–290. https://doi:10.1089/can.2018.0014

Giese, M.W., Lewis, M.A., & Smith, K.M. (2015). Development and Validation of a Reliable and Robust Method for the Analysis of Cannabinoids and Terpenes in Cannabis. J AOAC Int. 98(6), 1503–1522. https://doi: 10.5740/jaoacint.15–116

Gilbert, A.N. & Diverdi, J.A. (2018). Consumer perceptions of strain differences in Cannabis aroma. PLOS ONE. 13(2): e0192247. https://doi.org/10.1371/journal.pone.0192247

Ibrahim, E.A., Wang, M., Radwan, M.M., Wanas, A.S., Majumdar, C.G., Avula, B., Ikhlas, Y-H.W., Khan, A., Chandra, S., Lata, H., Hadad, G.M., Abdel Salam, R.A., Ibrahim, A.K., Ahmed, S.A., & Elsohly, M.A. (2019). Analysis of Terpenes in Cannabis sativa L. Using GC/MS: Method Development, Validation and Application. Planta Med 85(05), 431–438. https://doi: 10.1055/a-0828-8387

Jin, D., Jin, S., Yu, Y., Lee, C., & Chen, J. (2017). Classification of Cannabis Cultivars Marketed in Canada for Medical Purposes by Quantification of Cannabinoids and Terpenes Using HPLC-DAD and GC-MS. J Anal Bioanal Tech 8(1), 1000349. https://doi: 10.4172/2155-9872.1000349

Jou, Y.J., Hua, C.H., Lin, C.S., Wang, C.Y., Wan, L., Lin, Y.J., Huang, S.H., & Lin. C.W. (2016). Anticancer Activity of–Bisabolene in Human Neuroblastoma Cells via Induction of p53–Mediated Mitochondrial Apoptosis. Molecules. 21(5), 601–611. https://doi: 10.3390/molecules21050601

Jou, Y.J., Chen, C.J., Liu, Y.C., Way, T.D., Lai, C.H., Hua, C.H., Wang, C.Y., Huang, S.H., Kao, J.Y., & Lin, C.W. (2015). Quantitative phosphoproteomic analysis reveals bisabolene inducing p53–mediated apoptosis of human oral squamous cell carcinoma via HDAC2 inhibition and ERK1/2 activation. Proteomic, 15, 3296–3309. https://doi.org/10.1002/pmic.201400568

Kydryavtseva, A., Krasnov, G., Lipatova, A., Alekseev, B., Maganova, F., Shaposhnikov, M., Fedorova, M., Snezhkina, A., & Moskalev, A. (2016). Effects of Abies sibirica terpenes on cancer- and aging-associated pathways in human cells. Oncotarget. 7(50), 83744–83754. https://doi.org/10.18632/oncotarget.13467

Lia, Y., Kong, D., Fu, Y., Sussman, M.R., & Wu, H. (2020). The effect of developmental and environmental factors on secondary metabolites in medicinal plants. Plant Physiol. Biochem. 148, 80–89. https://doi.org/10.1016/j.plaphy.2020.01.006

Martin, A.M., Gomes, A.L., Boas, I.V., Marto, J., & Ribeiro, H.M. (2022). Cannabis-Based Products for the Treatment of Skin Inflammatory Diseases: A Timely Review. Pharmaceuticals. 15(2) 210. https://doi.org/10.3390/ph15020210

Menezes, I.O., Scherf, J.R., Martins, A.O.B.P.B., Ramos, A.G.B., Quintans, J.S.S., Coutinho, H.D.M., Filho, J.R., & Menezes, I.R.A. (2021). Biological properties of terpinolene evidenced by in silico, in vitro and in vivo studies: A systematic review. Phytomedicine. 93, 153768. https://doi.org/10.1016/j.phymed.2021.153768

Nuutinen, T. (2018). Medicinal properties of terpenes found in Cannabis sativa and Humulus lupulus. E. J. Med. Chem. 157, 198–228. http://doi.org/10.1016/j.ejmeth.2018.07.076

Omar, J., Oilvares, M., Amigo, J.M., & Etxebarria, N. (2014). Resolution of co-eluting compounds of Cannabis sativa in comprehensive two-dimensional gas chromatography/mass spectrometry detection with Multivariate Curve Resolution-Alternating Least Squares. Talanta 121, 273?280. http://doi.org/10.1016/j.talanta.2013.12.044

Porto, C.D., Decorti, D., & Natolino, A., (2014). Separation of aroma compounds from industrial hemp inflorescences (Cannabis sativa L.) by supercritical CO2 extraction and on–line fractionation. Ind. Crops Prod. 58, 99?103. https://doi.org/10.1016/j.indcrop.2014.03.042

Pussapaphan, N. & Busabong, W. (2022). Cannabis in the traditional Southern Thai medical recipe. Asian J Tred. Innov. Arts Textiles. 1(2), 31?61.

Quispe, L.S., Cruz, C.T., Choquelvica, Y.C., Quispe, A.S., Rodeiro, I., & Hernandez, I. (2019). Chemical composition and antibacterial activities of essential oil from Ageratina pentlandiana (DC.) R.M. King & H. Rob. Leaves grown in the Peruvian Andes. J. Essent. Oil Res. 28(3), 234–240. https://doi.org/10.1080/10412905.2015.1120691

Sommano, S.R., Chittasupho, C., Ruksiriwanich, W., & Jantrawut, P. (2020). The Cannabis Terpenes. Molecules.25, 5792. https://doi.org/10.3390/molecules25245792

Thielmann, J. & Muranyi, P. (2019). Review on the chemical composition of Litsea cubeba essential oils and the bioactivity of its major constituents citral and limonene. J. Essent. Oil Res. 31(5), 361–378. https://doi.org/10.1080/10412905.2019.1611671

UNODC–United Nations Office on Drugs and Crime. (2009). Recommended Method for the Identification and Analysis of Cannabis and Cannabis Products. United Nations, New York, USA, 60 p.

Yeo, S.K., Ali, A.Y., Hayward, O.A., Turnham, D., Jackson, T., Bowen, I.D., & Clarkson, R. (2016). b–Bisabolene, a sesquiterpene from the essential oil extract of opoponax (Commiphora guidottii), exhibits cytotoxicity in breast cancer cell lines. Phytother Res. 30, 418–425. https://doi.org/10.1002/ptr.5543

Zager, J.J., Lange, I., Srividya, N., Smith, A., & Lange, B.M. (2019). Gene Networks Underlying Cannabinoid and Terpenoid Accumulation in Cannabis. Plant Physiol. 180, 1877–1897. https://doi.org/10.1104/pp.18.01506