Advertisement

Metabolite Analysis of Cannabis sativa L. by NMR Spectroscopy

  • Isvett Josefina Flores-SanchezEmail author
  • Young Hae Choi
  • Robert Verpoorte
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 815)

Abstract

NMR-based metabolomics is an analytical platform, which has been used to classify and analyze Cannabis sativa L. cell suspension cultures and plants. Diverse groups of primary and secondary metabolites were identified by comparing NMR data with reference compounds and/or by structure elucidation using 1H-NMR, J-resolved, 1H-1H COSY, and 1H-13C HMBC spectroscopy. The direct extraction and the extraction by indirect fractionation are two suitable methods for the C. sativa sample preparation. Quantitative analyses could be performed without requiring fractionation or isolation procedures.

Key words

Cannabis sativa Metabolomics Multivariate data analysis Nuclear magnetic resonance Principal component analysis 

References

  1. 1.
    Flores-Sanchez IJ, Verpoorte R (2008) Secondary metabolism in cannabis. Phytochem Rev 7: 615–639CrossRefGoogle Scholar
  2. 2.
    Radwan MM, Ross SA, Slade D, et al (2008) Isolation and characterization of new cannabis constituents from a high potency variety. Planta Med 74: 267–272PubMedCrossRefGoogle Scholar
  3. 3.
    Radwan MM, ElSohly MA, Slade D, et al (2008) Non-cannabinoid constituents from a high potency Cannabis sativa variety. Phytochemistry 69: 2627–2633PubMedCrossRefGoogle Scholar
  4. 4.
    Debruyne D, Moulin M, Bigot Mc et al (1981) Identification and differentiation of resinous cannabinoid textile Cannabis: combined use of HPLC and high-resolution GLC. Bull Narc 33: 49–58PubMedGoogle Scholar
  5. 5.
    Ross SA, ElSohly HN, Elkashoury EA, et al (1996) Fatty acids of Cannabis seeds. Phytochem Anal 7: 279–283CrossRefGoogle Scholar
  6. 6.
    Hazekamp A, Simons R, Peltenburg-Looman A, et al (2004) Preparative isolation of cannabinoids from Cannabis sativa by centrifugal partition chromatography. J Liq Chromatogr Relat Technol 27: 2421–2439CrossRefGoogle Scholar
  7. 7.
    Raharjo TJ, Verpoorte R (2004) Methods for the analysis of cannabinoid in biological materials: a review. Phytochem Anal 15: 79–94PubMedCrossRefGoogle Scholar
  8. 8.
    Choi YH, Hazekamp A, Peltenburg-Looman A, et al (2004) NMR assignments of the major cannabinoids and cannabiflavonoids isolated from flowers of Cannabis sativa. Phytochem Anal 15: 345–354PubMedCrossRefGoogle Scholar
  9. 9.
    Ross SA, ElSohly MA, Sultana GNN, et al (2005) Flavonoid glycosides and cannabinoids from the pollen of Cannabis sativa L. Phytochem Anal 16: 45–48PubMedCrossRefGoogle Scholar
  10. 10.
    Hazekamp A, Peltenburg-Looman A, Verpoorte R, et al (2005) Chromatographic and spectroscopic data of cannabinoids from Cannabis sativa L. J Liq Chromatogr Relat Technol 28: 2361–2382CrossRefGoogle Scholar
  11. 11.
    Gutierrez A, Rodriguez IM, Del Rio JC (2006) Chemical characterization of lignin and lipid fractions in industrial hemp bast fibers used for manufacturing high-quality paper pulps. J Agric Food Chem 54: 2138–2144PubMedCrossRefGoogle Scholar
  12. 12.
    Flores-Sanchez IJ, Verpoorte R (2008) PKS activities and biosynthesis of cannabinoids and flavonoids in Cannabis sativa L. plants. Plant Cell Physiol 49: 1767–1782PubMedCrossRefGoogle Scholar
  13. 13.
    Holmes E, Tang H, Wang Y et al (2006) The assessment of plant metabolite profiles by NMR-based methodologies. Planta Med 72: 771–785PubMedCrossRefGoogle Scholar
  14. 14.
    Ward JL, Beale MH NMR spectroscopy in plant metabolomics. In Saito K, Dixon RA, Willmitzer L (ed) (2006) Plant Metabolomics, Biotechnology in Agriculture and Forestry, Vol. 57. Springer-Verlag, Berlin HeidelbergGoogle Scholar
  15. 15.
    Fiehn O (2002) Metabolomics: The link between genotypes and phenotypes. Plant Mol Biol 48: 155–171PubMedCrossRefGoogle Scholar
  16. 16.
    Sumner LW, Mendes P, Dixon RA (2003) Plant metabolomics: large-scale phytochemistry in the functional genomics era. Phytochemistry 62: 817–836PubMedCrossRefGoogle Scholar
  17. 17.
    Rochfort S (2005) Metabolomics reviewed: A new “omic” platform technology for systems biology and implications for natural products research. J Nat Prod 68: 1813–1820PubMedCrossRefGoogle Scholar
  18. 18.
    Kim HK, Verpoorte R (2010) Sample preparation for plant metabolomics. Phytochem Anal 21: 4–13PubMedCrossRefGoogle Scholar
  19. 19.
    Kim HK, Choi YH, Verpoorte R (2010) NMR-based metabolomic analysis of plants. Nat Protoc 5: 536–549PubMedCrossRefGoogle Scholar
  20. 20.
    Ludwing C, Viant MR (2010) Two-dimensional J-resolved NMR spectroscopy: Review of a key methodology in the metabolomics toolbox. Phytochem Anal 21: 22–32CrossRefGoogle Scholar
  21. 21.
    Choi YH, Kim HK, Hazekamp A, et al (2004) Metabolomic differentiation of Cannabis sativa cultivars using 1H-NMR spectroscopy and principal component analyses. J Nat Prod 67: 953–957PubMedCrossRefGoogle Scholar
  22. 22.
    Flores-Sanchez IJ, Pec J, Fei J, et al. (2009) Elicitation studies in cell suspension cultures of Cannabis sativa L. J Biotechnol 143: 157–168PubMedCrossRefGoogle Scholar
  23. 23.
    Pec J, Flores-Sanchez IJ, Choi YH et al (2010) Metabolic analysis of elicited cell suspension cultures of Cannabis sativa L. by 1H-NMR spectroscopy. Biotechnol Lett 32: 935–941PubMedCrossRefGoogle Scholar
  24. 24.
    Choi YH, Hazekamp A, Peltenburg-Looman AMG, et al. (2004) NMR assignments of the major cannabinoids isolated from flowers of Cannabis sativa. Phytochem Anal 15: 354–354Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Isvett Josefina Flores-Sanchez
    • 1
    Email author
  • Young Hae Choi
    • 2
  • Robert Verpoorte
    • 2
  1. 1.Institute of Biological ChemistryWashington State UniversityPullmanUSA
  2. 2.Pharmacognosy Department/Metabolomics, Gorlaeus Laboratories, Institute of BiologyLeiden UniversityRA LeidenThe Netherlands

Personalised recommendations