Analytical and Bioanalytical Chemistry

, Volume 407, Issue 17, pp 5125–5131 | Cite as

Quantitative profiling of endocannabinoids in lipoproteins by LC–MS/MS

  • Mesut Bilgin
  • Laura Bindila
  • Juergen Graessler
  • Andrej ShevchenkoEmail author
Research Paper
Part of the following topical collections:
  1. Lipidomics


Endocannabinoids belong to a diverse family of endogenous lipid bioregulators acting as physiological ligands of cannabinoid receptor type 1 and cannabinoid receptor type 2 in the central and peripheral nervous system. They are also present in nmol L−1 concentrations in human blood plasma; however, their association with possible molecular carriers remains poorly characterized. Here we report on the quantification of 46 endogenous molecular species from five major classes of endocannabinoids and endocannabinoid-related compounds in three lipoprotein fractions of human blood plasma: VLDL, LDL, HDL, and in the plasma lipoprotein-free fraction. Although sizable quantities of endocannabinoid-related molecules are associated with lipoproteins, we identified the lipoprotein-free fraction as a major carrier of endocannabinoids in blood circulation with the exception of 2-acylglycerols, which are markedly abundant in VLDL.


Endocannabinoids Lipoproteins Blood plasma LC–MS/MS Quantification 



The authors are grateful for all members of AS laboratory for their experimental support and useful discussions. We are particularly indebted to Dr. Susanne Sales (MPI CBG) for her expert assistance with data processing and Sigrid Nitzsche (TUD) for lipoprotein preparation. Work in AS laboratory was supported by TRR83 grant (project A17) from Deutsche Forschungsgemeinschaft (DFG).

Supplementary material

216_2015_8559_MOESM1_ESM.pdf (112 kb)
ESM 1 (PDF 111 kb)


  1. 1.
    Silvestri C, Di Marzo V (2013) The endocannabinoid system in energy homeostasis and the etiopathology of metabolic disorders. Cell Metab 17:475–490CrossRefGoogle Scholar
  2. 2.
    Kleberg K, Hassing HA, Hansen HS (2014) Classical endocannabinoid-like compounds and their regulation by nutrients. Biofactors 40:363–372Google Scholar
  3. 3.
    Hanus L, Shohami E, Bab I, Mechoulam R (2014) N-Acyl amino acids and their impact on biological processes. Biofactors 40:381–388Google Scholar
  4. 4.
    Pertwee RG, Howlett AC, Abood ME, Alexander SP, Di Marzo V, Elphick MR, Greasley PJ, Hansen HS, Kunos G, Mackie K, Mechoulam R, Ross RA (2010) International Union of Basic and Clinical Pharmacology. LXXIX. Cannabinoid receptors and their ligands: beyond CB(1) and CB(2). Pharmacol Rev 62:588–631CrossRefGoogle Scholar
  5. 5.
    Lerner R, Lutz B, Bindila L (2013) Tricks and tracks in the identification and quantification of endocannabinoids. eLS: 10.1002/9780470015902.a0023407
  6. 6.
    Astarita G, Geaga J, Ahmed F, Piomelli D (2009) Targeted lipidomics as a tool to investigate endocannabinoid function. Int Rev Neurobiol 85:35–55CrossRefGoogle Scholar
  7. 7.
    Balvers MG, Verhoeckx KC, Witkamp RF (2009) Development and validation of a quantitative method for the determination of 12 endocannabinoids and related compounds in human plasma using liquid chromatography-tandem mass spectrometry. J Chromatogr B Anal Technol Biomed Life Sci 877:1583–1590CrossRefGoogle Scholar
  8. 8.
    Balvers MG, Wortelboer HM, Witkamp RF, Verhoeckx KC (2013) Liquid chromatography-tandem mass spectrometry analysis of free and esterified fatty acid N-acyl ethanolamines in plasma and blood cells. Anal Biochem 434:275–283CrossRefGoogle Scholar
  9. 9.
    Astarita G, Piomelli D (2009) Lipidomic analysis of endocannabinoid metabolism in biological samples. J Chromatogr B Anal Technol Biomed Life Sci 877:2755–2767CrossRefGoogle Scholar
  10. 10.
    Lehtonen M, Storvik M, Malinen H, Hyytia P, Lakso M, Auriola S, Wong G, Callaway JC (2011) Determination of endocannabinoids in nematodes and human brain tissue by liquid chromatography electrospray ionization tandem mass spectrometry. J Chromatogr B Anal Technol Biomed Life Sci 879:677–694CrossRefGoogle Scholar
  11. 11.
    Skonberg C, Artmann A, Cornett C, Hansen SH, Hansen HS (2010) Pitfalls in the sample preparation and analysis of N-acylethanolamines. J Lipid Res 51:3062–3073CrossRefGoogle Scholar
  12. 12.
    De Petrocellis L, Cascio MG, Di Marzo V (2004) The endocannabinoid system: a general view and latest additions. Br J Pharmacol 141:765–774Google Scholar
  13. 13.
    Di Marzo V (2009) The endocannabinoid system: its general strategy of action, tools for its pharmacological manipulation and potential therapeutic exploitation. Pharmacol Res 60:77–84Google Scholar
  14. 14.
    Bjorklund E, Blomqvist A, Hedlin J, Persson E, Fowler CJ (2014) Involvement of fatty acid amide hydrolase and fatty acid binding protein 5 in the uptake of anandamide by cell lines with different levels of fatty acid amide hydrolase expression: a pharmacological study. PLoS One 9:e103479Google Scholar
  15. 15.
    Kaczocha M, Glaser ST, Deutsch DG (2009) Identification of intracellular carriers for the endocannabinoid anandamide. Proc Natl Acad Sci U S A 106:6375–6380Google Scholar
  16. 16.
    Khaliullina H, Bilgin M, Sampaio JL, Shevchenko A, Eaton S (2013) Lipoproteins carry endocannabinoids that inhibit the Hedgehog pathway. bioRxiv: doi: 10.1101/000570
  17. 17.
    Pietzsch J, Subat S, Nitzsche S, Leonhardt W, Schentke KU, Hanefeld M (1995) Very fast ultracentrifugation of serum lipoproteins: influence on lipoprotein separation and composition. Biochim Biophys Acta 1254:77–88CrossRefGoogle Scholar
  18. 18.
    Peterson AC, Russell JD, Bailey DJ, Westphall MS, Coon JJ (2012) Parallel reaction monitoring for high resolution and high mass accuracy quantitative, targeted proteomics. Mol Cell Proteomics 11:1475–1488CrossRefGoogle Scholar
  19. 19.
    Richardson D, Ortori CA, Chapman V, Kendall DA, Barrett DA (2007) Quantitative profiling of endocannabinoids and related compounds in rat brain using liquid chromatography-tandem electrospray ionization mass spectrometry. Anal Biochem 360:216–226CrossRefGoogle Scholar
  20. 20.
    Fanelli F, Di Lallo VD, Belluomo I, De Iasio R, Baccini M, Casadio E, Gasparini DI, Colavita M, Gambineri A, Grossi G, Vicennati V, Pasquali R, Pagotto U (2012) Estimation of reference intervals of five endocannabinoids and endocannabinoid related compounds in human plasma by two dimensional-LC/MS/MS. J Lipid Res 53:481–493CrossRefGoogle Scholar
  21. 21.
    Rouzer CA, Ghebreselasie K, Marnett LJ (2002) Chemical stability of 2-arachidonylglycerol under biological conditions. Chem Phys Lipids 119:69–82Google Scholar
  22. 22.
    Cifkova E, Holcapek M, Lisa M, Ovcacikova M, Lycka A, Lynen F, Sandra P (2012) Nontargeted quantitation of lipid classes using hydrophilic interaction liquid chromatography-electrospray ionization mass spectrometry with single internal standard and response factor approach. Anal Chem 84:10064–10070CrossRefGoogle Scholar
  23. 23.
    Buczynski MW, Svensson CI, Dumlao DS, Fitzsimmons BL, Shim JH, Scherbart TJ, Jacobsen FE, Hua XY, Yaksh TL, Dennis EA (2010) Inflammatory hyperalgesia induces essential bioactive lipid production in the spinal cord. J Neurochem 114:981–993Google Scholar
  24. 24.
    Sipe JC, Scott TM, Murray S, Harismendy O, Simon GM, Cravatt BF, Waalen J (2010) Biomarkers of endocannabinoid system activation in severe obesity. PLoS One 5:e8792Google Scholar
  25. 25.
    Bojesen IN, Hansen HS (2003) Binding of anandamide to bovine serum albumin. J Lipid Res 44:1790–1794CrossRefGoogle Scholar
  26. 26.
    Scherer M, Bottcher A, Liebisch G (2011) Lipid profiling of lipoproteins by electrospray ionization tandem mass spectrometry. Biochim Biophys Acta 1811:918–924CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Mesut Bilgin
    • 1
  • Laura Bindila
    • 2
  • Juergen Graessler
    • 3
  • Andrej Shevchenko
    • 1
    Email author
  1. 1.Max Planck Institute for Cell Biology and GeneticsDresdenGermany
  2. 2.Institute Physiological ChemistryUniversity Medical Center MainzMainzGermany
  3. 3.Department of Internal Medicine III, Medical Faculty Carl Gustav CarusTechnische Universität DresdenDresdenGermany

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