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In vitro stability of free and glucuronidated cannabinoids in urine following controlled smoked cannabis

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Abstract

Analyte stability is an important factor in urine test interpretation, yet cannabinoid stability data are limited. A comprehensive study of Δ9-tetrahydrocannabinol (THC), 11-hydroxy-THC (11-OH-THC), 11-nor-9-carboxy-THC (THCCOOH), cannabidiol, cannabinol, THC-glucuronide, and THCCOOH-glucuronide stabilities in authentic urine was completed. Urine samples after ad libitum cannabis smoking were pooled to prepare low and high pools for each study participant; baseline concentrations were measured within 24 h at room temperature (RT), 4 °C and -20 °C. Stability at RT, 4 °C and −20 °C was evaluated by Friedman tests for up to 1 year. THCCOOH, THC-glucuronide, and THCCOOH-glucuronide were quantified in baseline pools. RT THCCOOH baseline concentrations were significantly higher than −20 °C, but not 4 °C baseline concentrations. After 1 week at RT, THCCOOH increased, THCCOOH-glucuronide decreased, but THC-glucuronide was unchanged. In RT low pool, total THCCOOH (THCCOOH + THCCOOH-glucuronide) was significantly lower after 1 week. At 4 °C, THCCOOH was stable 2 weeks, THCCOOH-glucuronide 1 month and THC-glucuronide for at least 6 months. THCCOOH was stable frozen for 1 year, but 6 months high pool results were significantly higher than baseline; THC-glucuronide and THCCOOH-glucuronide were stable for 6 months. Total THCCOOH was stable 6 months at 4 °C, and frozen 6 months (low) and 1 year (high). THC, cannabidiol and cannabinol were never detected in urine; although not detected initially, 11-OH-THC was detected in 2 low and 3 high pools after 1 week at RT. Substantial THCCOOH-glucuronide deconjugation was observed at RT and 4 °C. Analysis should be conducted within 3 months if non-hydrolyzed THCCOOH or THCCOOH-glucuronide quantification is required.

Median (range) cannabinoid stability in baseline positive pools after 1 week at room temperature and up to 26 and 52 weeks at 4 °C and −20 °C, respectively, in low and high cannabinoid urine pools, collected after controlled cannabis smoking. Significant differences from baseline for *low pool and #high pool. Dashed lines represents ±20 % for THCCOOH (analyte with a deuterated internal standard) and ±30 % for THC-glucuronide, THCCOOH-glucuronide and total THCCOOH (analytes without a matched deuterated internal standard)

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References

  1. United Nations Office on Drugs and Crime (UNODC) (2013) World Drug Report 2013. http://www.unodc.org/unodc/secured/wdr/wdr2013/World_Drug_Report_2013.pdf. Accessed 21 September 2013

  2. Substance Abuse and Mental Health Services Administration (2012) Results from the 2011 National Survey on Drug Use and Health: Summary of National Findings. NSDUH Series H-44. U.S. Department of Health and Human Services, Rockville. http://www.samhsa.gov/data/NSDUH/2012SummNatFindDetTables/NationalFindings/NSDUHresults2012.htm. Accessed 21 September 2013.

  3. Drummer OH, Kourtis I, Beyer J, Tayler P, Boorman M, Gerostamoulos D (2012) The prevalence of drugs in injured drivers. Forensic Sci Int 215:14–17

    Article  CAS  Google Scholar 

  4. Lacey JH, Kelley-Baker T, Furr-Holden D, Voas RB, Romano E, Ramirez A, Brainard K, Moore C, Torres P, Berning A (2009) 2007 National Roadside Survey of Alcohol and Drug Use by Drivers: Drug Results. National Highway Traffic Safety Administration Office of Behavioral Safety Research. http://www.nhtsa.gov/Driving+Safety/Research+&+Evaluation/ci.2007+National+Roadside+Survey+of+Alcohol+and+Drug+Use+by+Drivers.print. Accessed 21 September 2013.

  5. Mazur A, Lichti CF, Prather PL, Zielinska AK, Bratton SM, Gallus-Zawada A, Finel M, Miller GP, Radomińska-Pandya A, Moran JH (2009) Characterization of Human Hepatic and Extrahepatic UDP-Glucuronosyltransferase Enzymes Involved in the Metabolism of Classic Cannabinoids. Drug Metab Dispos 37(7):1496–1504

    Article  CAS  Google Scholar 

  6. Lowe R, Abraham T, Darwin W, Herning R, Cadet J, Huestis M (2009) Extended Urinary delta 9 Tetrahydrocannabinol Excretion in Chronic Cannabis Users Precludes Use as a Biomarker of New Drug Exposure. Drug Alcohol Depen 105(1–2):24–32

    Article  CAS  Google Scholar 

  7. Goodwin RS, Darwin WD, Chiang CN, Shih M, Li S-H, Huestis MA (2008) Urinary Elimination of 11-Nor-9-Carboxy-D9-tetrahydrocannabinol in Cannabis Users During Continuously Monitored Abstinence. J Anal Toxicol 32(8):562–566

    Article  CAS  Google Scholar 

  8. Smith ML, Barnes AJ, Huestis MA (2009) Identifying New Cannabis Use with Urine Creatinine-Normalized THCCOOH Concentrations and Time Intervals Between Specimen Collections. J Anal Toxicol 33(4):185–189

    Article  CAS  Google Scholar 

  9. Schwilke EW, Gullberg RG, Darwin WD, Chiang CN, Cadet JL, Gorelick DA, Pope HG, Huestis MA (2011) Differentiating new cannabis use from residual urinary cannabinoid excretion in chronic, daily cannabis users. Addiction 106(3):499–506

    Article  Google Scholar 

  10. Daldrup T, Meininger I (1998) In: Berghaus G, Kruger HP (eds) Cannabis im Strassenverkehr. Gustav Fischer Verlag, Stuttgart

    Google Scholar 

  11. Skopp G, Pötsch L, Ganßmann B, Mauden M, Richter B, Aderjan R, Mattern R (1999) Freie und glucuronidierte Cannabinoide im Urin – Untersuchungen zur Einschätzung des Konsumverhaltens. Rechtsmedizin 10(1):21–28

    Article  Google Scholar 

  12. Jung J, Kempf J, Mahler H, Weinmann W (2007) Detection of Delta9-tetrahydrocannabinolic acid A in human urine and blood serum by LC-MS/MS. J Mass Spectrom 42(3):354–360

    Article  CAS  Google Scholar 

  13. Mareck U, Haenelt N, Geyer H, Guddat S, Kamber M, Brenneisen R, Thevis M, Schänzer W (2009) Temporal indication of cannabis use by means of THC glucuronide determination. Drug Test Anal 1(11–12):505–510

    Article  CAS  Google Scholar 

  14. Blanc JA, Manneh VA, Ernst R, Berger DE, de Keczer SA, Chase C, Centofanti JM, DeLizza AJ (1993) Adsorption losses from urine-based cannabinoid calibrators during routine use. Clin Chem 39:1705–1717

    CAS  Google Scholar 

  15. Roth KDW, Siegel NA, Johnson RW, Litauszki L, Salvati L Jr, Harrington CA, Wray LK (1996) Investigation of the effects of solution composition and container material type on the loss of 11-nor-delta-9-THC-9-carboxylic acid. J Anal Toxicol 20:291–300

    Article  CAS  Google Scholar 

  16. Stout PR, Horn CK, Lesser DR (2000) Loss of THCCOOH from urine specimens stored in polypropylene and polyethylene containers at different temperatures. J Anal Toxicol 24(7):567–571

    Article  CAS  Google Scholar 

  17. Jamerson MH, McCue JJ, Klette KL (2005) Urine pH, container composition, and exposure time influence adsorptive loss of 11-nor-delta9-tetrahydrocannabinol-9-carboxylic acid. J Anal Toxicol 29(7):627–631

    Article  CAS  Google Scholar 

  18. Dugan S, Bogema S, Schwartz RW, Lappas NT (1994) Stability of drugs of abuse in urine samples stored at -20 degrees. J Anal Toxicol 18(7):391–396

    Article  CAS  Google Scholar 

  19. Golding Fraga S, Diaz-Flores Estevez J, Diaz Romero C (1998) Stability of cannabinoids in urine in three storage temperatures. Ann Clin Lab Sci 28(3):160–162

    CAS  Google Scholar 

  20. Romberg RW, Past MR (1994) Reanalysis of forensic urine specimens containing benzoylecgonine and THC-COOH. J Forensic Sci 39:479–485

    CAS  Google Scholar 

  21. Skopp G, Potsch L (2002) Stability of 11-nor-delta(9)-carboxy-tetrahydrocannabinol glucuronide in plasma and urine assessed by liquid chromatography-tandem mass spectrometry. Clin Chem 48(2):301–306

    CAS  Google Scholar 

  22. Skopp G, Potsch L (2004) An investigation of the stability of free and glucuronidated 11-nor-delta-9-tetrahydrocannabinol-9-carboxylic acid in authentic urine samples. J Ana Toxicol 28:35–40

    Article  CAS  Google Scholar 

  23. Felli M, Martello S, Chiarotti M (2011) LC–MS–MS method for simultaneous determination of THCCOOH and THCCOOH-glucuronide in urine: Application to workplace confirmation tests. Forensic Sci Int 204(1–3):67–73

    Article  CAS  Google Scholar 

  24. Scheidweiler KB, Desrosiers NA, Huestis MA (2012) Simultaneous quantification of free and glucuronidated cannabinoids in human urine by liquid chromatography tandem mass spectrometry. Clin Chim Acta 413(23–24):1839–1847

    Article  CAS  Google Scholar 

  25. Scheidweiler KB, Schwope DM, Karschner EL, Desrosiers NA, Gorelick DA, Huestis MA (2013) In Vitro Stability of Free and Glucuronidated Cannabinoids in Blood and Plasma Following Controlled Smoked Cannabis. Clin Chem 59:1108–1117

    Article  CAS  Google Scholar 

  26. Faed EM (1984) Properties of acyl glucuronides: implications for studies of the pharmacokinetics and metabolism of acidic drugs. Drug Metab Rev 15(5–6):1213–1249

    Article  CAS  Google Scholar 

  27. Shipkova M, Armstrong VW, Oellerich M, Wieland E (2003) Acyl glucuronide drug metabolites: toxicological and analytical implications. Ther Drug Monit 25(1):1–16

    Article  CAS  Google Scholar 

  28. Robertson LW, Koh SW, Huff SR, Malhotra RK, Ghosh A (1978) Microbiological oxidation of the pentyl side chain of cannabinoids. Experientia 34(8):1020–1022

    Article  CAS  Google Scholar 

  29. Abraham TT, Lowe RH, Pirnay SO, Darwin WD, Huestis MA (2007) Simultaneous GC-EI-MS determination of delta9-tetrahydrocannabinol, 11-hydroxy-delta9-tetrahydrocannabinol, and 11-nor-9-carboxy-delta9-tetrahydrocannabinol in human urine following tandem enzyme-alkaline hydrolysis. J Anal Toxicol 31(8):477–485

    Article  CAS  Google Scholar 

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Acknowledgments

We acknowledge the contributions of the clinical staffs of the National Institute on Drug Abuse, Intramural Research Program, and Behavioral Pharmacology Research Unit and Clinical Research Unit, Johns Hopkins Bayview Medical Center, as well as Dr. David M. Schwope for protocol assistance, Dan Nichols and the staff at the Forensic Toxicology Drug Testing Laboratory in Fort Meade who provided urine creatinine data, the Graduate Partnership Program, NIH and the “Fondation Baxter et Alma Ricard”. This research was funded by the Intramural Research Program, National Institute on Drug Abuse, NIH.

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Authors and Affiliations

  1. Chemistry and Drug Metabolism, Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, 251 Bayview Boulevard, Suite 200 Room 05A-721, Baltimore, MD, 21224, USA

    Nathalie A. Desrosiers, Dayong Lee, Karl B. Scheidweiler, Marta Concheiro-Guisan, David A. Gorelick & Marilyn A. Huestis

  2. Program in Toxicology, University of Maryland Baltimore, 655 W. Baltimore Street, Baltimore, MD, 21201, USA

    Nathalie A. Desrosiers & Dayong Lee

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  1. Nathalie A. Desrosiers
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  2. Dayong Lee
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  3. Karl B. Scheidweiler
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  6. Marilyn A. Huestis
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Correspondence to Marilyn A. Huestis.

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Desrosiers, N.A., Lee, D., Scheidweiler, K.B. et al. In vitro stability of free and glucuronidated cannabinoids in urine following controlled smoked cannabis. Anal Bioanal Chem 406, 785–792 (2014). https://doi.org/10.1007/s00216-013-7524-7

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