GC-MS Metabolite Profile and Identification of Unusual Homologous Cannabinoids in High Potency Cannabis sativa

Abstract

Phytochemical investigation of the lipids extracted from seeds of Cannabis sativa by GC-MS showed 43 cannabinoids, 16 of which are new. The extract is dominated by Δ⁹-tetrahydrocannabinolic acid (A) and its neutral derivative trans-Δ⁹-tetrahydrocannabinol-C₅ (THC) Cis and trans-Δ⁹-tetrahydrocannabinol-C₇ isomers with an ethyl-pentyl branched chain together with minor amounts of trans-Δ⁹-tetrahydrocannabinol with a methyl-pentyl C₆ branched side chain were identified as new natural compounds. Four cannabichromene isomers with a C₅ side chain are postulated to be derived from the double bond migration at the terminal isoprenyl unit. C₇ cannabichromene together with the neutral and acidic forms of cannabinol-C₇ were also detected. The mass spectrum of these homologues as trimethylsilyl (TMS) derivatives are presented, and the fragmentation patterns are discussed.

Key words

Cannabinoids - Cannabis sativa - Cannabaceae - phytochemical analysis - GC-MS

• References

• 1 Clarke RC, Watson DP. Cannabis and natural Cannabis Medicines. In: El-Sohly MA. ed. Marijuana and the Cannabinoids. Totowa, New Jersey: Humana Press Inc; 2007: 1-16
• 2 Mechoulam R. Cannabinoids as Therapeutics. Basel: Virkhäuser; 2005: 1-264
• 3 Radwan MM, Wanas AS, Chandra S, El-Sohly MA. Natural Cannabinoids of Cannabis and Methods of Analysis. In: Chandra S, Lata H, El-Sohly MA. eds. Cannabis sativa L. – Botany and Biotechnology. Cham, Switzerland: Springer International Publishing AG; 2017: 161-182
• 4 United Nation Office on Drugs and Crime (UNODC). Cannabis: a short review. Published 2012. Available at: https://www.unodc.org/documents/drug-prevention-and-treatment/cannabis_review.pdf Accessed September 28, 2019
• 5 El-Sohly MA, Radwan MM, Gul W, Chandra S, Galal A. Phytochemistry of Cannabis sativa L. In: Kinghorn A, Falk H, Gibbons S, Kobayashi J. eds. Phytocannabinoids. Progress in the Chemistry of organic natural Products, Volume 103. Cham, Switzerland: Springer International Publishing AG; 2017: 1-36
• 6 Shoyama Y, Yagi M, Nishioka I, Yamautchi T. Biosynthesis of cannabinoid acids. Phytochemistry 1975; 14: 2189-2192
• 7 Veress T, Szanto JI, Leisztner L. Determination of cannabinoid acids by high-performance liquid chromatography of their neutral derivatives formed by thermal decarboxylation. J Chromatogr A 1990; 520: 339-347
• 8 Razdan RK. Structure-activity relationships in cannabinoids. Pharmacol Rev 1986; 38: 75-149
• 9 Fetterman PS, Keith ES, Waller CW, Guerrero O, Doorenbos NJ, Quimby MW. Mississippi-grown Cannabis sativa L.: Preliminary observation on chemical definition of phenotype and variations in tetrahydrocannabinol content versus age, sex, and plant part. J of Pharm Sci 1971; 60: 1246-1249
• 10 Billets S, El-Feraly F, Fetterman PS, Turner CE. Constituents of Cannabis sativa L. XII-Mass spectral fragmentation patterns for some cannabinoid acids as their TMS derivatives. Org Mass Spectrom 1976; 11: 741-751
• 11 Hanus LO. Pharmacological and therapeutic secrets of plant and brain (endo)cannabinoids. Med Res Rev 2009; 29: 213-271
• 12 Harvey DJ. Mass spectrometry of the cannabinoids and their metabolites. Mass Spectrom Rev 1987; 6: 135-229
• 13 Moore M, Rickards RW, Ronneberg H. Cannabinoid studies. IV. Stereoselective and regiospecific syntheses of (±)-Δ9-trans- and (±)-Δ9-cis-6a,10a-tetrahydrocannabinol. Aust J Chem 1984; 37: 2339-2348
• 14 Lewis R, Ward S, Johnson R, Thorburn Burns D. Distribution of the principal cannabinoids within bars of compressed cannabis resin. Anal Chim Acta 2005; 538: 399-405
• 15 Gaoni Y, Mechoulam R. Isolation, structure, and partial synthesis of an active constituent of hashish. J Am Chem Soc 1964; 86: 1646-1647
• 16 Harvey DJ. The mass spectra of the trimethylsilyl derivatives of Δ1- and Δ6-tetrahydrocannabinol. Biomed Mass Spectrom 1981; 8: 575-578
• 17 Vree TB. Mass spectrometry of cannabinoids. J Pharm Sci 1977; 66: 1444-1450
• 18 Choi YH, Hazekamp A, Peltenburg-Looman AMG, Frédérich M, Erkelens C, Lefeber AWM, Verpoorte R. NMR assignments of the major cannabinoids and cannabiflavonoids isolated from flowers of Cannabis sativa . Phytochem Anal 2004; 15: 345-354
• 19 Brown NK, Harvey DJ. Metabolism of n-hexyl-homologues of delta-8-tetrahydrocannabinol and delta-9-tetrahydrocannabinol in the mouse. Eur J Drug Metab Ph 1988; 13: 165-176
• 20 Goad LJ, Hammam ASA, Dennis A, Goodwin TW. Biosynthesis of the phytosterol side chain. Nature 1966; 210: 1322-1324
• 21 Nes WD, Song Z, Dennis AL, Zhou W, Nam J, Miller MB. Biosynthesis of phytosterols kinetic mechanism for the enzymatic C-methylation of sterols. J Biol Chem 2003; 278: 34505-34516
• 22 Groeneveld HW. Tracing steroid synthesis in plants. Crit Rev Biochem Mol 1999; 34: 59-69
• 23 Harvey DJ. Cyclic alkylboronates as derivatives for the characterization of cannabinolic acids by combined gas chromatography and mass spectrometry. Biomed Mass Spectrom 1977; 4: 88-93
• 24 Harvey DJ. Stability of cannabinoids in dried samples of cannabis dating from around 1896–1905. J Ethnopharmacol 1990; 28: 117-128
• 25 Harvey DJ, Brown NK. Electron impact-induced fragmentation of the trimethylsilyl derivatives of monohydroxy-hexahydrocannabinols. Biol Mass Spectrom 1991; 20: 292-302
• 26 Turner CE, El-Sohly MA. Biological activity of cannabichromene, its homologs and isomers. J Clin Pharmacol 1981; 21: 283-291
• 27 Ross SA, El-Sohly MA, Sultana GNN, Mehmedic Z, Hossain ChF, Chandra S. Flavonoid glycosides and cannabinoids from the pollen of Cannabis sativa L. Phytochem Anal 2005; 16: 45-48
• 28 Hazekamp A, Peltenburg A, Verpoorte R, Giroud C. Chromatographic and spectroscopic data of cannabinoids from Cannabis sativa L. J. Liq Chromatogr Relat Technol 2005; 28: 2361-2382
• 29 El-Sohly MA, Slade D. Chemical constituents of marijuana: The complex mixture of natural cannabinoids. Life Sci 2005; 78: 539-548
• 30 Adams RP. Identification of essential Oil Components by Gas Chromatography/Mass Spectrometry, 4th edition. Illinois: Allured Publishing Corporation; 2007
• 31 Degenhardt F, Stehle F, Kayser O. The Biosynthesis of Cannabinoids. In: Preedy V. ed. Handbook of Cannabis and related Pathologies. London: Elsevier; 2017: 13-23
• 32 Mechoulam R, Yagen B. Stereoselective cyclization of cannabinoid 1,5 diene. Tetrahedron Lett 1969; 60: 5349-5352
• 33 Hanus LO, Levy R, De La Vega D, Katz L, Roman M, Tomícek P. The main cannabinoids content in hashish samples seized in Israel and Czech Republic. Israel J. Plant Sci 2016; 63: 182-190
• 34 Harvey DJ, Brown NK. A method for the structural determination of cannabichromene metabolites by mass spectrometry. Rapid Commun Mass Sp 1990; 4: 135-136
• 35 Taura F, Morimoto S, Shoyama Y. Cannabinerolic acid, a cannabinoid from Cannabis sativa . Phytochemistry 1995; 39: 457-458
• 36 Fellermeier M, Zenk MH. Prenylation of olivetolate by a hemp transferase yields cannabigerolic acid, the precursor of tetrahydrocannabinol. FEBS Lett 1998; 427: 283-285
• 37 Gaoni Y, Mechoulam R. The structure and synthesis of cannabigerol, a new hashish constituent. Proc Chem Soc 1964; 82
• 38 Radwan M, Ross S, Slade D, Ahmed S, Zulfiqar F, El-Sohly M. Isolation and characterization of new cannabis constituents from a high potency variety. Planta Med 2008; 74: 267-272
• 39 Cardenia V, Toschi TG, Scappini S, Rubino RC, Rodriguez-Estrada MT. Development and validation of a fast gas chromatography/mass spectrometry method for the determination of cannabinoids in Cannabis sativa L. J Food Drug Anal 2018; 4: 1283-1292
• 40 Budzikiewicz H, Alpin RT, Lightner DA, Djerassi C, Mechoulam R, Gaoni Y. Massenspektroskopie und ihre Anwendung auf strukturelle und stereochemische Probleme-LXVIII. Massenspektroskopische Untersuchung der Inhaltstoffe von Haschisch. Tetrahedron 1965; 21: 1881-1888
• 41 Kinzer GW, Foltz RL, Mitchell RI, Battelle. Truitt EB. The fate of the cannabinoid components of marihuana during smoking. Bull Narcot 1974; 26: 41-54
• 42 El-Sohly MA, El-Feraly FS, Turner CE. Isolation and characterization of (+)-cannabitriol and (−)-10-ethoxy-9-hydroxy-Δ6a(10a)-tetrahydrocannabinol: two new cannabinoids from Cannabis sativa L. extract. Lloydia 1977; 40: 275-280
• 43 Roth N, Wohlfarth A, Auwärter V. Regioselective synthesis of THCA-A and THCA-B by reaction of Δ9-THC with magnesium methyl carbonate (MMC). Toxichem Krimtech 2011; 78: 316-319
• 44 Harvey DJ, Brown NK. Identification of cannabichromene metabolites by mass spectrometry: identification of eight new dihydroxy metabolites in the rabbit. Biol Mass Spectrom 1991; 20: 275-285
• 45 Lee YR, Xia L. Efficient one-pot synthetic approaches for cannabinoid analogues and their application to biologically interesting (−)-hexahydrocannabinol and (+)-hexahydrocannabinol. Tetrahedron Lett 2008; 49: 3283-3287