Analytical and Bioanalytical Chemistry

, Volume 408, Issue 23, pp 6461–6471 | Cite as

Simultaneous quantification of 11 cannabinoids and metabolites in human urine by liquid chromatography tandem mass spectrometry using WAX-S tips

  • Maria Andersson
  • Karl B. ScheidweilerEmail author
  • Cristina Sempio
  • Allan J. Barnes
  • Marilyn A. Huestis
Research Paper


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.


Cannabinoid Urine LC-MS/MS DPX THC 













9-Tetrahydrocannabinolic acid













This research was funded by the Intramural Research Program (IRP) of the National Institute on Drug Abuse, National Institutes of Health.

Compliance with ethical standards

Conflict of interest

The authors declare no conflicts of interest.


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Copyright information

© Springer-Verlag Berlin Heidelberg (outside the USA) 2016

Authors and Affiliations

  • Maria Andersson
    • 1
  • Karl B. Scheidweiler
    • 1
    Email author
  • Cristina Sempio
    • 1
  • Allan J. Barnes
    • 1
  • Marilyn A. Huestis
    • 1
    • 2
  1. 1.Chemistry and Drug Metabolism Section, Clinical Pharmacology and Therapeutics Research Branch, Intramural Research Program, National Institute on Drug AbuseNational Institutes of HealthBaltimoreUSA
  2. 2.University of Maryland School of MedicineBaltimoreUSA

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