Download PDF
Planta Med 2018; 84(04): 260-266
DOI: 10.1055/s-0043-124873
Natural Product Chemistry and Analytical Studies
Original Papers
Georg Thieme Verlag KG Stuttgart · New York

Quantitative Determination of Δ9-THC, CBG, CBD, Their Acid Precursors and Five Other Neutral Cannabinoids by UHPLC-UV-MS

Yan-Hong Wang
1   National Center for Natural Products Research, Research Institute of Pharmaceutical Sciences, School of Pharmacy, The University of Mississippi, University, MS, USA
,
Bharathi Avula
1   National Center for Natural Products Research, Research Institute of Pharmaceutical Sciences, School of Pharmacy, The University of Mississippi, University, MS, USA
,
Mahmoud A. ElSohly
1   National Center for Natural Products Research, Research Institute of Pharmaceutical Sciences, School of Pharmacy, The University of Mississippi, University, MS, USA
2   Department of Pharmaceutics and Drug Delivery, School of Pharmacy, The University of Mississippi, University, MS, USA
,
Mohamed M. Radwan
1   National Center for Natural Products Research, Research Institute of Pharmaceutical Sciences, School of Pharmacy, The University of Mississippi, University, MS, USA
,
Mei Wang
1   National Center for Natural Products Research, Research Institute of Pharmaceutical Sciences, School of Pharmacy, The University of Mississippi, University, MS, USA
,
Amira S. Wanas
1   National Center for Natural Products Research, Research Institute of Pharmaceutical Sciences, School of Pharmacy, The University of Mississippi, University, MS, USA
,
Zlatko Mehmedic
1   National Center for Natural Products Research, Research Institute of Pharmaceutical Sciences, School of Pharmacy, The University of Mississippi, University, MS, USA
,
Ikhlas A. Khan
1   National Center for Natural Products Research, Research Institute of Pharmaceutical Sciences, School of Pharmacy, The University of Mississippi, University, MS, USA
3   Division of Pharmacognosy, Department of BioMolecular Sciences, School of Pharmacy, The University of Mississippi, University, MS, USA
› Author Affiliations
Further Information

Publication History

received 05 January 2017
revised 29 November 2017

accepted 07 December 2017

Publication Date:
20 December 2017 (online)

   Permissions and Reprints

Abstract

Cannabinoids are a group of terpenophenolic compounds in the medicinal plant Cannabis sativa (Cannabaceae family). Cannabigerolic acid, Δ9-tetrahydrocannabinolic acid A, cannabidiolic acid, Δ9-tetrahydrocannabinol, cannabigerol, cannabidiol, cannabichromene, and tetrahydrocannabivarin are major metabolites in the classification of different strains of C. sativa. Degradation or artifact cannabinoids cannabinol, cannabicyclol, and Δ8-tetrahydrocannabinol are formed under the influence of heat and light during processing and storage of the plant sample. An ultrahigh-performance liquid chromatographic method coupled with photodiode array and single quadruple mass spectrometry detectors was developed and validated for quantitative determination of 11 cannabinoids in different C. sativa samples. Compounds 1 – 11 were baseline separated with an acetonitrile (with 0.05% formic acid) and water (with 0.05% formic acid) gradient at a flow rate of 0.25 mL/min on a Waters Cortec UPLC C18 column (100 mm × 2.1 mm I. D., 1.6 µm). The limits of detection and limits of quantitation of the 11 cannabinoids were below 0.2 and 0.5 µg/mL, respectively. The relative standard deviation for the precision test was below 2.4%. A mixture of acetonitrile and methanol (80 : 20, v/v) was proven to be the best solvent system for the sample preparation. The recovery of all analytes was in the range of 97 – 105%. A total of 32 Cannabis samples including hashish, leaves, and flower buds were analyzed.

Key words

Cannabis sativa - Cannabaceae - cannabinoids - tetrahydrocannabinol - cannabidiol - tetrahydrocannabinolic acid - UHPLC-UV-MS
 

  • References

  • 1 Abrams DI, Hilton JF, Leiser RJ, Shade SB, Elbeik TA, Aweeka FT, Benowitz NL, Bredt BM, Kosel B, Aberg JA, Deeks SG, Mitchell TF, Mulligan K, Bacchetti P, McCune JM, Schambelan M. Short-term effects of cannabinoids in patients with HIV-1 infection: a randomized, placebo-controlled clinical trial. Ann Intern Med 2003; 139: 258-266
    CrossrefPubMedGoogle Scholar
  • 2 Jatoi A, Windschitl HE, Loprinzi CL, Sloan JA, Dakhil SR, Mailliard JA, Pundaleeka S, Kardinal CG, Fitch TR, Krook JE, Novotny PJ, Christensen B. Dronabinol versus megestrol acetate versus combination therapy for cancer-associated anorexia: a North Central Cancer Treatment Group Study. J Clin Oncol 2002; 20: 567-573
    CrossrefPubMedGoogle Scholar
  • 3 Aizpurua-Olaizola O, Soydaner U, Ozturk E, Schibano D, Simsir Y, Navarro P, Etxebarria N, Usobiaga A. Evolution of the cannabinoid and terpene content during the growth of Cannabis sativa plants from different chemotypes. J Nat Prod 2016; 79: 324-331
    CrossrefPubMedGoogle Scholar
  • 4 Beal JE, Olson R, Laubenstein L, Morales JO, Bellman P, Yangco B, Lefkowitz L, Plasse TF, Shepard KV. Dronabinol as a treatment for anorexia associated with weight loss in patients with AIDS. J Pain Symptom Manage 1995; 10: 89-97
    CrossrefPubMedGoogle Scholar
  • 5 De Backer B, Maebe K, Verstraete AG, Charlier C. Evolution of the content of THC and other major cannabinoids in drug-type cannabis cuttings and seedlings during growth of plants. J Forensic Sci 2012; 57: 918-922
    CrossrefPubMedGoogle Scholar
  • 6 ElSohly MA, Gul W. Constituents of Cannabis sativa . In: Pertwee RG. ed. Handbook of Cannabis. Oxford: Oxford University Press; 2014: 3-22
    CrossrefGoogle Scholar
  • 7 De Backer B, Debrus B, Lebrun P, Theunis L, Dubois N, Decock L, Verstraete A, Hubert P, Charlier C. Innovative development and validation of an HPLC/DAD method for the qualitative and quantitative determination of major cannabinoids in cannabis plant material. J Chromatogr B: Anal Technol Biomed Life Sci 2009; 877: 4115-4124
    CrossrefPubMedGoogle Scholar
  • 8 Sirikantaramas S, Morimoto S, Shoyama Y, Ishikawa Y, Wada Y, Shoyama Y, Taura F. The gene controlling marijuana psychoactivity: molecular cloning and heterologous expression of Delta1-tetrahydrocannabinolic acid synthase from Cannabis sativa L. J Biol Chem 2004; 279: 39767-39774
    CrossrefPubMedGoogle Scholar
  • 9 Cascini F, Boschi I. Chapter e1. Tetrahydrocannabinol Concentration and genetic Characterization of Cannabis. In: Preedy VR. ed. Handbook of Cannabis and related Pathologies: Biology, Pharmacology, Diagnosis, and Treatment. Cambridge: Academic Press; 2017: e1-e10
    Google Scholar
  • 10 Giese MW, Lewis MA, Giese L, Smith KM. Development and validation of a reliable and robust method for the analysis of cannabinoids and terpenes in cannabis. J AOAC Int 2015; 98: 1503-1522
    CrossrefPubMedGoogle Scholar
  • 11 Ross SA, ElSohly MA. The volatile oil composition of fresh and air-dried buds of Cannabis sativa L. J Nat Prod 1996; 59: 49-51
    CrossrefPubMedGoogle Scholar
  • 12 De Brabanter N, Van Gansbeke W, Hooghe F, Van Eenoo P. Fast quantification of 11-nor-Δ9-tetrahydrocannabinol-9-carboxylic acid (THCA) using microwave-accelerated derivatisation and gas chromatography-triple quadrupole mass spectrometry. Forensic Sci Int 2013; 224: 90-95
    CrossrefPubMedGoogle Scholar
  • 13 Ross SA, Mehmedic Z, Murphy TP, ElSohly MA. GC-MS analysis of the total Δ9-THC content of both drug- and fiber-type Cannabis seeds. J Anal Toxicol 2000; 24: 715-717
    CrossrefPubMedGoogle Scholar
  • 14 Gul W, Gul SW, Radwan MM, Wanas AS, Mehmedic Z, Khan II, Sharaf MHM, El Sohly MA. Determination of 11 cannabinoids in biomass and extracts of different varieties of Cannabis using high-performance liquid chromatography. J AOAC Int 2015; 98: 1523-1528
    CrossrefPubMedGoogle Scholar
  • 15 Chang CW, Tung CW, Tsai CC, Wu YT, Hsu MC. Determination of cannabinoids in hemp nuts products in Taiwan by HPLC-MS/MS coupled with chemometric analysis: quality evaluation and a pilot human study. Drug Test Anal 2017; 9: 888-897
    CrossrefPubMedGoogle Scholar
  • 16 Citti C, Ciccarella G, Braghiroli D, Parenti C, Vandelli MA, Cannazza G. Medicinal cannabis: Principal cannabinoids concentration and their stability evaluated by a high performance liquid chromatography coupled to diode array and quadrupole time of flight mass spectrometry method. J Pharm Biomed Anal 2016; 128: 201-209
    CrossrefPubMedGoogle Scholar
  • 17 Aizpurua-Olaizola O, Omar J, Navarro P, Olivares M, Etxebarria N, Usobiaga A. Identification and quantification of cannabinoids in Cannabis sativa L. plants by high performance liquid chromatography-mass spectrometry. Anal Bioanal Chem 2014; 406: 7549-7560
    CrossrefPubMedGoogle Scholar
  • 18 Salomone A, Gerace E, DʼUrso F, Di Corcia D, Vincenti M. Simultaneous analysis of several synthetic cannabinoids, THC, CBD and CBN, in hair by ultra-high performance liquid chromatography tandem mass spectrometry. Method validation and application to real samples. J Mass Spectrom 2012; 47: 604-610
    CrossrefPubMedGoogle Scholar
  • 19 Valen A, Leere AM, Strand DH, Skari R, Berg T. Determination of 21 drugs in oral fluid using fully automated supported liquid extraction and UHPLC-MS/MS. Drug Test Anal 2017; 9: 808-823
    CrossrefPubMedGoogle Scholar
  • 20 Wang M, Wang YH, Avula B, Radwan MM, Wanas AS, Mehmedic Z, van Antwerp J, ElSohly MA, Khan IA. Quantitative determination of cannabinoids in cannabis and cannabis products using ultra-high-performance supercritical fluid chromatography and diode array/mass spectrometric detection. J Forensic Sci 2017; 62: 602-611
    CrossrefPubMedGoogle Scholar
  • 21 Ambach L, Penitschka F, Broillet A, Konig S, Weinmann W, Bernhard W. Simultaneous quantification of delta-9-THC, THC-acid A, CBN and CBD in seized drugs using HPLC-DAD. Forensic Sci Int 2014; 243: 107-111
    CrossrefPubMedGoogle Scholar
  • 22 Ferioli V, Rustichelli C, Pavesi G, Gamberini G. Analytical characterization of hashish samples. Chromatographia 2000; 52: 39-44
    CrossrefPubMedGoogle Scholar
  • 23 ICH. Validation of analytical procedures: text and methodology. ICH Harmonized Triparitite Guideline. Available at http://www.ich.org/fileadmin/Public_Web_Site/ICH_Products/Guidelines/Quality/Q2_R1/Step4/Q2_R1__Guideline.pdf Accessed June 12, 2017
    PubMedGoogle Scholar