Skip to main content

Advertisement

SpringerLink
Log in

Development of a rapid column-switching LC-MS/MS method for the quantification of THCCOOH and THCCOOH-glucuronide in whole blood for assessing cannabis consumption frequency

  • Research Paper
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

Abstract

The concentration of 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THCCOOH) in whole blood is used as a parameter for assessing the consumption behavior of cannabis consumers. The blood level of THCCOOH-glucuronide might provide additional information about the frequency of cannabis use. To verify this assumption, a column-switching liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the rapid and direct quantification of free and glucuronidated THCCOOH in human whole blood was newly developed. The method comprised protein precipitation, followed by injection of the processed sample onto a trapping column and subsequent gradient elution to an analytical column for separation and detection. The total LC run time was 4.5 min. Detection of the analytes was accomplished by electrospray ionization in positive ion mode and selected reaction monitoring using a triple-stage quadrupole mass spectrometer. The method was fully validated by evaluating the following parameters: linearity, lower limit of quantification, accuracy and imprecision, selectivity, extraction efficiency, matrix effect, carry-over, dilution integrity, analyte stability, and re-injection reproducibility. All acceptance criteria were analyzed and the predefined criteria met. Linearity ranged from 5.0 to 500 μg/L for both analytes. The method was successfully applied to whole blood samples from a large collective of cannabis consumers, demonstrating its applicability in the forensic field.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. United Nations Office on Drugs and Crime (UNODC) (2015) World drug report 2015. http://www.unodc.org/documents/wdr2015/World_Drug_Report_2015.pdf. Accessed 28 Sept 2015.

  2. European Monitoring Centre for Drugs and Drug Addiction (EMCDDA) (2012) Driving under the influence of drugs, alcohol and medicines in Europe—findings from the DRUID project. http://www.emcdda.europa.eu/attachements.cfm/att_192773_EN_TDXA12006ENN.pdf. Accessed 08 Oct 2015.

  3. Lacey JH, Kelley-Baker T, Voas RB, Romano E, Furr-Holden CD, Torres P, et al. Alcohol- and drug-involved driving in the United States: methodology for the 2007 National Roadside Survey. Eval Rev. 2011;35:319–53. doi:10.1177/0193841x11422446.

    Article  Google Scholar 

  4. Beasley EE, Beirness DJ (2012) Alcohol and drug use among drivers following the introduction of immediate roadside prohibitions in British Columbia: findings from the 2012 roadside survey. http://www.pssg.gov.bc.ca/osmv/shareddocs/bc-roadside-report2012.pdf. Accessed 08 Oct 2015

  5. Davey J, Armstrong K, Martin P. Results of the Queensland 2007-2012 roadside drug testing program: the prevalence of three illicit drugs. Accid Anal Prev. 2014;65:11–7. doi:10.1016/j.aap.2013.12.007.

    Article  Google Scholar 

  6. Hartman RL, Huestis MA. Cannabis effects on driving skills. Clin Chem. 2013;59:478–92. doi:10.1373/clinchem.2012.194381.

    Article  CAS  Google Scholar 

  7. Battistella G, Fornari E, Thomas A, Mall JF, Chtioui H, Appenzeller M, et al. Weed or wheel! FMRI, behavioural, and toxicological investigations of how cannabis smoking affects skills necessary for driving. PLoS ONE. 2013;8, e52545. doi:10.1371/journal.pone.0052545.

    Article  CAS  Google Scholar 

  8. Asbridge M, Mann R, Cusimano MD, Trayling C, Roerecke M, Tallon JM, et al. Cannabis and traffic collision risk: findings from a case-crossover study of injured drivers presenting to emergency departments. Int J Public Health. 2014;59:395–404. doi:10.1007/s00038-013-0512-z.

    Article  Google Scholar 

  9. Desrosiers NA, Ramaekers JG, Chauchard E, Gorelick DA, Huestis MA. Smoked cannabis’ psychomotor and neurocognitive effects in occasional and frequent smokers. J Anal Toxicol. 2015;39:251–61. doi:10.1093/jat/bkv012.

    Article  Google Scholar 

  10. Wong K, Brady J, Li G. Establishing legal limits for driving under the influence of marijuana. Inj Epidemiol. 2014;1:26. doi:10.1186/s40621-014-0026-z.

    Article  Google Scholar 

  11. Huestis MA. Human cannabinoid pharmacokinetics. Chem Biodivers. 2007;4:1770–804. doi:10.1002/cbdv.200790152.

    Article  CAS  Google Scholar 

  12. Kelly P, Jones RT. Metabolism of tetrahydrocannabinol in frequent and infrequent marijuana users. J Anal Toxicol. 1992;16:228–35. doi:10.1093/jat/16.4.228.

    Article  CAS  Google Scholar 

  13. Toennes SW, Ramaekers JG, Theunissen EL, Moeller MR, Kauert GF. Comparison of cannabinoid pharmacokinetic properties in occasional and heavy users smoking a marijuana or placebo joint. J Anal Toxicol. 2008;32:470–7. doi:10.1093/jat/32.7.470.

    Article  CAS  Google Scholar 

  14. Skopp G, Pötsch L. Cannabinoid concentrations in spot serum samples 24-48 hours after discontinuation of cannabis smoking. J Anal Toxicol. 2008;32:160–4. doi:10.1093/jat/32.2.160.

    Article  CAS  Google Scholar 

  15. Desrosiers NA, Himes SK, Scheidweiler KB, Concheiro-Guisan M, Gorelick DA, Huestis MA. Phase I and II cannabinoid disposition in blood and plasma of occasional and frequent smokers following controlled smoked cannabis. Clin Chem. 2014;60:631–43. doi:10.1373/clinchem.2013.216507.

    Article  CAS  Google Scholar 

  16. Daldrup T, Käferstein H, Köhler H, Maier R-D, Musshoff F. Deciding between one off/occasional and regular cannabis consumption. Blutalkohol. 2000;37:39–47.

    CAS  Google Scholar 

  17. Fabritius M, Favrat B, Chtioui H, Battistella G, Annoni JM, Appenzeller M, et al. THCCOOH concentrations in whole blood: are they useful in discriminating occasional from heavy smokers? Drug Test Anal. 2014;6:155–63. doi:10.1002/dta.1581.

    Article  CAS  Google Scholar 

  18. Fabritius M, Augsburger M, Chtioui H, Favrat B, Giroud C. Fitness to drive and cannabis: validation of two blood THCCOOH thresholds to distinguish occasional users from heavy smokers. Forensic Sci Int. 2014;242:1–8. doi:10.1016/j.forsciint.2014.05.014.

    Article  CAS  Google Scholar 

  19. Schwope DM, Karschner EL, Gorelick DA, Huestis MA. Identification of recent cannabis use: whole-blood and plasma free and glucuronidated cannabinoid pharmacokinetics following controlled smoked cannabis administration. Clin Chem. 2011;57:1406–14. doi:10.1373/clinchem.2011.171777.

    Article  CAS  Google Scholar 

  20. Wall ME, Taylor HL. Conjugation of acidic metabolites of Δ-8 and Δ-9-THC in man. In: Harvey DJ, editor. Marihuana ’84. Proceedings of the Oxford Symposium on Cannabis. Oxford: IRL Press; 1985. p. 69–76.

    Google Scholar 

  21. Mazur A, Lichti CF, Prather PL, Zielinska AK, Bratton SM, Gallus-Zawada A, et al. Characterization of human hepatic and extrahepatic UDP-glucuronosyltransferase enzymes involved in the metabolism of classic cannabinoids. Drug Metab Dispos. 2009;37:1496–504. doi:10.1124/dmd.109.026898.

    Article  CAS  Google Scholar 

  22. Guillemette C. Pharmacogenomics of human UDP-glucuronosyltransferase enzymes. Pharmacogenomics J. 2003;3:136–58. doi:10.1038/sj.tpj.6500171.

    Article  CAS  Google Scholar 

  23. Staub C. Chromatographic procedures for determination of cannabinoids in biological samples, with special attention to blood and alternative matrices like hair, saliva, sweat and meconium. J Chromatogr B Biomed Sci Appl. 1999;733:119–26. doi:10.1016/S0378-4347(99)00249-2.

    Article  CAS  Google Scholar 

  24. Fabritius M, Staub C, Giroud C. Analyse des cannabinoïdes par spectrométrie de masse en mode tandem. Ann Toxicol Anal. 2011;23:21–35. doi:10.1051/ata/2011104.

    Article  CAS  Google Scholar 

  25. Schwope DM, Scheidweiler KB, Huestis MA. Direct quantification of cannabinoids and cannabinoid glucuronides in whole blood by liquid chromatography-tandem mass spectrometry. Anal Bioanal Chem. 2011;401:1273–83. doi:10.1007/s00216-011-5197-7.

    Article  CAS  Google Scholar 

  26. Gomes RL, Meredith W, Snape CE, Sephton MA. Analysis of conjugated steroid androgens: deconjugation, derivatisation and associated issues. J Pharm Biomed Anal. 2009;49:1133–40. doi:10.1016/j.jpba.2009.01.027.

    Article  CAS  Google Scholar 

  27. Schwilke EW, Schwope DM, Karschner EL, Lowe RH, Darwin WD, Kelly DL, et al. Delta9-tetrahydrocannabinol (THC), 11-hydroxy-THC, and 11-nor-9-carboxy-THC plasma pharmacokinetics during and after continuous high-dose oral THC. Clin Chem. 2009;55:2180–9. doi:10.1373/clinchem.2008.122119.

    Article  CAS  Google Scholar 

  28. Kemp PM, Abukhalaf IK, Manno JE, Manno BR, Alford DD, McWilliams ME, et al. Cannabinoids in humans. II. The influence of three methods of hydrolysis on the concentration of THC and two metabolites in urine. J Anal Toxicol. 1995;19:292–8. doi:10.1093/jat/19.5.292.

    Article  CAS  Google Scholar 

  29. Fabritius M, Staub C, Mangin P, Giroud C. Analysis of cannabinoids in oral fluid by liquid chromatography–tandem mass spectrometry. Forensic Toxicol. 2013;31:151–63. doi:10.1007/s11419-012-0168-z.

    Article  CAS  Google Scholar 

  30. Skopp G, Pötsch L. Stability of 11-nor-delta(9)-carboxy-tetrahydrocannabinol glucuronide in plasma and urine assessed by liquid chromatography-tandem mass spectrometry. Clin Chem. 2002;48:301–6.

    CAS  Google Scholar 

  31. Gronewold A, Skopp G. A preliminary investigation on the distribution of cannabinoids in man. Forensic Sci Int. 2011;210:e7–11. doi:10.1016/j.forsciint.2011.04.010.

    Article  CAS  Google Scholar 

  32. Scheidweiler KB, Desrosiers NA, Huestis MA. Simultaneous quantification of free and glucuronidated cannabinoids in human urine by liquid chromatography tandem mass spectrometry. Clin Chim Acta. 2012;413:1839–47. doi:10.1016/j.cca.2012.06.034.

    Article  CAS  Google Scholar 

  33. Felli M, Martello S, Chiarotti M. LC-MS-MS method for simultaneous determination of THCCOOH and THCCOOH-glucuronide in urine: application to workplace confirmation tests. Forensic Sci Int. 2011;204:67–73. doi:10.1016/j.forsciint.2010.05.004.

    Article  CAS  Google Scholar 

  34. Food and Drug Administration (2001) Guidance for industry: bioanalytical method validation. http://www.fda.gov/downloads/Drugs/…/Guidances/ucm070107.pdf. Accessed 28 Sept 2015.

  35. European Medicines Agency (2011) Guideline on bioanalytical method validation. http://www.ema.europa.eu/docs/en_GB/document_library/Scientific_guideline/2011/08/WC500109686.pdf. Accessed 28 Sept 2015.

  36. Almeida AM, Castel-Branco MM, Falcão AC. Linear regression for calibration lines revisited: weighting schemes for bioanalytical methods. J Chromatogr B Anal Technol Biomed Life Sci. 2002;774:215–22. doi:10.1016/S1570-0232(02)00244-1.

    Article  CAS  Google Scholar 

  37. König S, Aebi B, Lanz S, Gasser M, Weinmann W. On-line SPE LC-MS/MS for the quantification of delta9-tetrahydrocannabinol (THC) and its two major metabolites in human peripheral blood by liquid chromatography tandem mass spectrometry. Anal Bioanal Chem. 2011;400:9–16. doi:10.1007/s00216-011-4708-x.

    Article  Google Scholar 

  38. Brunetto Mdel R, Contreras Y, Delgado Y, Gallignani M, Estela JM, Martin VC. Development and validation of a rapid column-switching high-performance liquid chromatographic method for the determination of lamotrigine in human serum. J Chromatogr Sci. 2009;47:478–84. doi:10.1093/chromsci/47.6.478.

    Article  Google Scholar 

  39. Wickremsinhe ER, Lee LB, Schmalz CA, Torchia J, Ruterbories KJ. High sensitive assay employing column switching chromatography to enable simultaneous quantification of an amide prodrug of gemcitabine (LY2334737), gemcitabine, and its metabolite dFdU in human plasma by LC-MS/MS. J Chromatogr B Anal Technol Biomed Life Sci. 2013;932:117–22. doi:10.1016/j.jchromb.2013.06.008.

    Article  CAS  Google Scholar 

  40. Satinsky D, Havlikova L, Solich P. HPLC column-switching technique for sample preparation and fluorescence determination of propranolol in urine using fused-core columns in both dimensions. Anal Bioanal Chem. 2013;405:6583–7. doi:10.1007/s00216-013-7098-4.

    Article  CAS  Google Scholar 

  41. Alves MN, Zanchetti G, Piccinotti A, Tameni S, De Martinis BS, Polettini A. Determination of cocaine and metabolites in hair by column-switching LC-MS-MS analysis. Anal Bioanal Chem. 2013;405:6299–306. doi:10.1007/s00216-013-7046-3.

    Article  CAS  Google Scholar 

  42. Scheidweiler KB, Schwope DM, Karschner EL, Desrosiers NA, Gorelick DA, Huestis MA. In vitro stability of free and glucuronidated cannabinoids in blood and plasma following controlled smoked cannabis. Clin Chem. 2013;59:1108–17. doi:10.1373/clinchem.2012.201467.

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors acknowledge the staff of the Forensic Toxicology laboratory for their technical assistance.

Author information

Authors and Affiliations

  1. Institute of Forensic Medicine, University of Bern, Bühlstrasse 20, 3012, Bern, Switzerland

    Marianne Hädener, Wolfgang Weinmann & Stefan König

  2. Department of Chemistry and Biochemistry, University of Bern, Freiestrasse 3, 3012, Bern, Switzerland

    Stefan Schürch

Authors
  1. Marianne Hädener
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Wolfgang Weinmann
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Stefan Schürch
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Stefan König
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Marianne Hädener.

Ethics declarations

Conflict of interest

The authors declare that they have no competing interests.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Hädener, M., Weinmann, W., Schürch, S. et al. Development of a rapid column-switching LC-MS/MS method for the quantification of THCCOOH and THCCOOH-glucuronide in whole blood for assessing cannabis consumption frequency. Anal Bioanal Chem 408, 1953–1962 (2016). https://doi.org/10.1007/s00216-016-9311-8

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-016-9311-8

Keywords

Search

Navigation