A comprehensive cannabinoid urine quantification method may improve clinical and forensic result interpretation and is necessary to support our clinical research. A liquid chromatography tandem mass spectrometry quantification method for ∆9-tetrahydrocannabinol (THC), 11-hydroxy-THC (11-OH-THC), 11-nor-9-carboxy-THC (THCCOOH), ∆9-tetrahydrocannabinolic acid (THCAA), cannabinol (CBN), cannabidiol (CBD), cannabigerol (CBG), ∆9-tetrahydrocannabivarin (THCV), 11-nor-9-carboxy-THCV (THCVCOOH), THC-glucuronide (THC-gluc), and THCCOOH-glucuronide (THCCOOH-gluc) in urine was developed and validated according to the Scientific Working Group on Toxicology guidelines. Sample preparation consisted of disposable pipette extraction (WAX-S) of 200 μL urine. Separation was achieved on a Kinetex C18 column using gradient elution with flow rate 0.5 mL/min, mobile phase A (10 mM ammonium acetate in water), and mobile phase B (15 % methanol in acetonitrile). Total run time was 14 min. Analytes were monitored in both positive and negative ionization modes by scheduled multiple reaction monitoring. Linear ranges were 0.5–100 μg/L for THC and THCCOOH; 0.5–50 μg/L for 11-OH-THC, CBD, CBN, THCAA, and THC-gluc; 1–100 μg/L for CBG, THCV, and THCVCOOH; and 5–500 μg/L for THCCOOH-gluc (R 2 > 0.99). Analytical biases were 88.3–113.7 %, imprecisions 3.3–14.3 %, extraction efficiencies 42.4–81.5 %, and matrix effect −10 to 32.5 %. We developed and validated a comprehensive, simple, and rapid LC-MS/MS cannabinoid urine method for quantification of 11 cannabinoids and metabolites. This method is being used in a controlled cannabis administration study, investigating urine cannabinoid markers documenting recent cannabis use, chronic frequent smoking, or route of drug administration and potentially improving urine cannabinoid result interpretation.
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Musshoff F, Madea B. Review of biologic matrices (urine, blood, hair) as indicators of recent or ongoing cannabis use. Ther Drug Monit. 2006;28(2):155–63.
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(23–24):1839–47.
Desrosiers NA, Lee D, Scheidweiler KB, Concheiro-Guisan M, Gorelick DA, Huestis MA. In vitro stability of free and glucuronidated cannabinoids in urine following controlled smoked cannabis. Anal Bioanal Chem. 2014;406(3):785–92.
United Nations Office on Drugs and Crime. World drug report 2015. 2015. https://www.unodc.org/documents/wdr2015/World_Drug_Report_2015.pdf. Accessed 17 June 2016.
ElSohly MA, Mehmedic Z, Foster S, Gon C, Chandra S, Church JC. Changes in cannabis potency over the last 2 decades (1995–2014): analysis of current data in the United States. Biol Psychiatry. 2016;79(7):613–9.
Cressey D. The cannabis experiment. Nature. 2015;524(7565):280–3.
European Monitoring Centre for Drugs and Drug Addiction. EMCDDA highlights new health risks as products and patterns of use change. 2016. http://www.emcdda.europa.eu/news/2016/6/european-drug-report-2016-highlights. Accessed 17 June 2016.
Hartman RL, Huestis MA. Cannabis effects on driving skills. Clin Chem. 2013;59(3):478–92.
Scheidweiler KB, Cone EJ, Moolchan ET, Huestis MA. Dose-related distribution of codeine, cocaine, and metabolites into human hair following controlled oral codeine and subcutaneous cocaine administration. J Pharmacol Exp Ther. 2005;313(2):909–15.
Polettini A, Cone EJ, Gorelick DA, Huestis MA. Incorporation of methamphetamine and amphetamine in human hair following controlled oral methamphetamine administration. Anal Chim Acta. 2012;726:35–43.
Schwilke EW, Gullberg RG, Darwin WD, Chiang CN, Cadet JL, Gorelick DA, et al. Differentiating new cannabis use from residual urinary cannabinoid excretion in chronic, daily cannabis users. Addiction. 2011;106(3):499–506.
Meyer MR. Trends in analyzing emerging drugs of abuse—from seized samples to body samples. Anal Bioanal Chem. 2014;406(25):6105–10.
Reisfield GM, Wasan AD, Jamison RN. The prevalence and significance of cannabis use in patients prescribed chronic opioid therapy: a review of the extant literature. Pain Med. 2009;10(8):1434–41.
Macdonald S, Hall W, Roman P, Stockwell T, Coghlan M, Nesvaag S. Testing for cannabis in the work-place: a review of the evidence. Addiction. 2010;105(3):408–16.
Dams R, Huestis MA, Lambert WE, Murphy CM. Matrix effect in bio-analysis of illicit drugs with LC-MS/MS: influence of ionization type, sample preparation, and biofluid. J Am Soc Mass Spectrom. 2003;14(11):1290–4.
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(7):1496–504.
Lowe RH, Abraham TT, Darwin WD, Herning R, Cadet JL, Huestis MA. Extended urinary Delta9-tetrahydrocannabinol excretion in chronic cannabis users precludes use as a biomarker of new drug exposure. Drug Alcohol Depend. 2009;105(1–2):24–32.
Goodwin RS, Darwin WD, Chiang CN, Shih M, Li SH, Huestis MA. Urinary elimination of 11-nor-9-carboxy-delta9-tetrahydrocannnabinol in cannabis users during continuously monitored abstinence. J Anal Toxicol. 2008;32(8):562–9.
Scientific Working Group for Forensic T. Scientific Working Group for Forensic Toxicology (SWGTOX) standard practices for method validation in forensic toxicology. J Anal Toxicol. 2013;37(7):452–74.
Matuszewski BK, Constanzer ML, Chavez-Eng CM. Strategies for the assessment of matrix effect in quantitative bioanalytical methods based on HPLC-MS/MS. Anal Chem. 2003;75(13):3019–30.
Desrosiers NA, Scheidweiler KB, Huestis MA. Quantification of six cannabinoids and metabolites in oral fluid by liquid chromatography-tandem mass spectrometry. Drug Test Anal. 2015;7(8):684–94.
Hidvegi E, Somogyi GP. Detection of cannabigerol and its presumptive metabolite in human urine after Cannabis consumption. Pharmazie. 2010;65(6):408–11.
Radunz L, Westphal F, Maser E, Rochholz G. THCVA-A—a new additional marker for illegal cannabis consumption. Forensic Sci Int. 2012;215(1–3):171–4.
Raikos N, Schmid H, Nussbaumer S, Ambach L, Lanz S, Langin A, et al. Determination of Delta9-tetrahydrocannabinolic acid A (Delta9-THCA-A) in whole blood and plasma by LC-MS/MS and application in authentic samples from drivers suspected of driving under the influence of cannabis. Forensic Sci Int. 2014;243:130–6.
Jung J, Kempf J, Mahler H, Weinmann W. Detection of Delta9-tetrahydrocannabinolic acid A in human urine and blood serum by LC-MS/MS. J Mass Spectrom. 2007;42(3):354–60.
Moore C, Rana S, Coulter C. Simultaneous identification of 2-carboxy-tetrahydrocannabinol, tetrahydrocannabinol, cannabinol and cannabidiol in oral fluid. J Chromatogr B Analyt Technol Biomed Life Sci. 2007;852(1–2):459–64.
Fabritius M, Chtioui H, Battistella G, Annoni JM, Dao K, Favrat B, et al. Comparison of cannabinoid concentrations in oral fluid and whole blood between occasional and regular cannabis smokers prior to and after smoking a cannabis joint. Anal Bioanal Chem. 2013;405(30):9791–803.
Moosmann B, Roth N, Auwarter V. Hair analysis for THCA-A, THC and CBN after passive in vivo exposure to marijuana smoke. Drug Test Anal. 2014;6(1–2):119–25.
Moosmann B, Roth N, Auwarter V. Hair analysis for Delta(9)-tetrahydrocannabinolic acid A (THCA-A) and Delta(9)-tetrahydrocannabinol (THC) after handling cannabis plant material. Drug Test Anal. 2016;8(1):128–32.
Englund A, Atakan Z, Kralj A, Tunstall N, Murray R, Morrison P. The effect of five day dosing with THCV on THC-induced cognitive, psychological and physiological effects in healthy male human volunteers: a placebo-controlled, double-blind, crossover pilot trial. J Psychopharmacol. 2016;30(2):140–51.
Wargent ET, Zaibi MS, Silvestri C, Hislop DC, Stocker CJ, Stott CG, et al. The cannabinoid Delta(9)-tetrahydrocannabivarin (THCV) ameliorates insulin sensitivity in two mouse models of obesity. Nutr Diabetes. 2013;3:e68.
ElSohly MA, deWit H, Wachtel SR, Feng S, Murphy TP. Delta9-tetrahydrocannabivarin as a marker for the ingestion of marijuana versus Marinol: results of a clinical study. J Anal Toxicol. 2001;25(7):565–71.
Schroeder JL, Marinetti LJ, Smith RK, Brewer WE, Clelland BL, Morgan SL. The analysis of delta9-tetrahydrocannabinol and metabolite in whole blood and 11-nor-delta9-tetrahydrocannabinol-9-carboxylic acid in urine using disposable pipette extraction with confirmation and quantification by gas chromatography–mass spectrometry. J Anal Toxicol. 2008;32(8):659–66.
Ellison ST, Brewer WE, Morgan SL. Comprehensive analysis of drugs of abuse in urine using disposable pipette extraction. J Anal Toxicol. 2009;33(7):356–65.
Wei B, Wang L, Blount BC. Analysis of cannabinoids and their metabolites in human urine. Anal Chem. 2015;87(20):10183–7.
This research was funded by the Intramural Research Program (IRP) of the National Institute on Drug Abuse, National Institutes of Health.
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The authors declare no conflicts of interest.
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Andersson, M., Scheidweiler, K.B., Sempio, C. et al. Simultaneous quantification of 11 cannabinoids and metabolites in human urine by liquid chromatography tandem mass spectrometry using WAX-S tips. Anal Bioanal Chem 408, 6461–6471 (2016). https://doi.org/10.1007/s00216-016-9765-8