EFFECT OF THIDIAZURON ON CANNABIS SATIVA L. PLANTLETS AND ISOLATION OF CANNABINOIDS USING CENTRIFUGAL PARTITION CHROMATOGRAPHY

Main Article Content

Pathamaporn Pathompak
Tossaton Charoonratana
Thanapat Songsak

Abstract

Cannabis (Cannabis sativa L.) is a plant native to Thailand. The ability of cannabis to exert effects on health varies depending on different amounts of the active compounds, cannabinoids. These compounds accumulate in the inflorescence of the female plant. When the plant is grown from seed, it is susceptible to genetic instability. However, growing clones using tissue culture guarantees the desired genetics. This study examined the effects of the plant growth regulator thidiazuron (TDZ) on growth and cannabinoids in cloned cannabis plantlets. Moreover, the primary cannabinoids, ∆9-tetrahydrocannabinol (THC) and cannabidiol (CBD) isolation from seized 400 g of dried cannabis were optimized using Centrifugal Partition Chromatography (CPC), and they were used as the reference standard for compounds quantification. The result showed that 1 g of cannabis extract provided 50 mg CBD and 20 mg THC within 2 h. Moreover, it was found that at TDZ 1.0 µM, the plantlets produced the highest number of multiple shoots after 4 weeks, while the cannabinoids profile under UV detector (220 nm) was no different compared to control without TDZ supplemented. No difference was found in the amounts of CBD and THC from both groups; CBD and THC content was around 0.12 and 0.05 %w/w, respectively, in the plantlets supplemented with 1.0 µM TDZ, while CBD and THC content was around 0.13 and 0.05 % w/w, respectively, in the control group. Treatment with 1.0 µM TDZ can be used to boost cannabis clones. This provides an alternative cloning method for commercial cannabis growers.

Article Details

How to Cite
1.
Pathompak P, Charoonratana T, Songsak T. EFFECT OF THIDIAZURON ON CANNABIS SATIVA L. PLANTLETS AND ISOLATION OF CANNABINOIDS USING CENTRIFUGAL PARTITION CHROMATOGRAPHY. Interprof J Health Sci [Internet]. 2021 May 18 [cited 2024 Dec. 6];19(1):1-8. Available from: https://li05.tci-thaijo.org/index.php/IJHS/article/view/41
Section
Research Articles

References

Chokevivat V. 2019. Cannabis: is it really a magic medicine?. J. Thai Tradit. Altern. Med. 17(2): 324-40.

Erland LAE, Giebelhaus RT, Victor JMR, Murch SJ, Saxena PK. 2020. The morphoregulatory role of thidiazuron: Metabolomics-guided hypothesis generation for mechanisms of activity. Biomolecules 10(9): 1253. Doi: 10.3390/biom10091253.

Hazekamp A, Simons R, Peltenburg‐Looman A, Sengers M, van Zweden R, Verpoorte R. 2004. Preparative isolation of cannabinoids from Cannabis sativa by Centrifugal Partition Chromatography. J. Liq. Chromatogr. Relat. Technol. 27(15): 2421–39.

Huetteman CA, Preece JE. 1993. Thidiazuron: a potent cytokinin for woody plant tissue culture. Plant Cell Tissue Organ Cult. 33(2): 105–19.

Lata H, Chandra S, Khan I, ElSohly MA. 2009. Thidiazuron-induced high-frequency direct shoot organogenesis of Cannabis sativa L. In Vitro Cell. Dev. Biol. Plant 45(1): 12–9.

Lata H, Chandra S, Khan IA, ElSohly MA. 2017. Micropropagation of Cannabis sativa L.—An Update. Cannabis Sativa L. - Botany and Biotechnology, 285–97.

Mander L, Liu H. 2010. Comprehensive Natural Products II: Chemistry and Biology (1st ed.). Elsevier Science.

Mandrioli M, Tura M, Scotti S, Toschi TG. 2019. Fast detection of 10 cannabinoids by RP-HPLC-UV method in Cannabis sativa L. Molecules 24(11): 2113. Doi: 10.3390/molecules24112113.

McRae G, Melanson JE. 2020. Quantitative determination and validation of 17 cannabinoids in cannabis and hemp using liquid chromatography-tandem mass spectrometry. Anal. Bioanal. Chem. 412: 7381-93.

Pertwee RG. 2006. Cannabinoid pharmacology: the first 66 years. Br. J. Pharmacol. 147(S1), S163–71.

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. Rev. Bras. Farmacogn. 28; 669-72.

Transcribe the inscription of King Jayavarman VII found “Auspicious drug court”. (Accessed on Oct. 8, 2020, at http://www.yclsakhon.com/index.php?lay=show&ac=article&Id=539367372&fbclid=IwAR1uUIYHStLDuemkN59lTfJG8a4lfS3RbA-ZGo4s1vTpC1RV0eAgFuEwIn0/)

Wang R, He L, Xia B, Tong J, Li N, Peng F. 2009. A micropropagation system for cloning of hemp (Cannabis sativa L.) by shoot tip culture. Pak. J. Bot. 41(2), 603-8.

Wróbel T, Dreger M, Wielgus K, Słomski R. 2017. The application of plant in vitro cultures in cannabinoid production. Biotechnol. Lett. 40(3), 445–54.