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Validation of a liquid chromatography tandem mass spectrometry (LC-MS/MS) method to detect cannabinoids in whole blood and breath

  • Hubbard, Jacqueline A.1
  • Smith, Breland E.2
  • Sobolesky, Philip M.3
  • Kim, Sollip4
  • Hoffman, Melissa A.1
  • Stone, Judith5
  • Huestis, Marilyn A.6
  • Grelotti, David J.7
  • Grant, Igor7
  • Marcotte, Thomas D.7
  • Fitzgerald, Robert L.1
  • 1 University of California, CA 92121 , (United States)
  • 2 Insource Diagnostics, CA , (United States)
  • 3 Santa Clara Valley Medical Center, CA , (United States)
  • 4 Inje University Ilsan Paik Hospital, Ilsan Seo-gu, Republic of Korea , (South Korea)
  • 5 San Francisco Medical Center, Laboratory Medicine, Parnassus Chemistry, CA , (United States)
  • 6 Thomas Jefferson University, PA , (United States)
  • 7 University of California, CA , (United States)
Published Article
Clinical Chemistry and Laboratory Medicine (CCLM)
Walter de Gruyter GmbH
Publication Date
Sep 17, 2019
DOI: 10.1515/cclm-2019-0600
De Gruyter


Background The widespread availability of cannabis raises concerns regarding its effect on driving performance and operation of complex equipment. Currently, there are no established safe driving limits regarding ∆9-tetrahydrocannabinol (THC) concentrations in blood or breath. Daily cannabis users build up a large body burden of THC with residual excretion for days or weeks after the start of abstinence. Therefore, it is critical to have a sensitive and specific analytical assay that quantifies THC, the main psychoactive component of cannabis, and multiple metabolites to improve interpretation of cannabinoids in blood; some analytes may indicate recent use. Methods A liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed to quantify THC, cannabinol (CBN), cannabidiol (CBD), 11-hydroxy-THC (11-OH-THC), (±)-11-nor-9-carboxy-Δ9-THC (THCCOOH), (+)-11-nor-Δ9-THC-9-carboxylic acid glucuronide (THCCOOH-gluc), cannabigerol (CBG), and tetrahydrocannabivarin (THCV) in whole blood (WB). WB samples were prepared by solid-phase extraction (SPE) and quantified by LC-MS/MS. A rapid and simple method involving methanol elution of THC in breath collected in SensAbues® devices was optimized. Results Lower limits of quantification ranged from 0.5 to 2 μg/L in WB. An LLOQ of 80 pg/pad was achieved for THC concentrations in breath. Calibration curves were linear (R2>0.995) with calibrator concentrations within ±15% of their target and quality control (QC) bias and imprecision ≤15%. No major matrix effects or drug interferences were observed. Conclusions The methods were robust and adequately quantified cannabinoids in biological blood and breath samples. These methods will be used to identify cannabinoid concentrations in an upcoming study of the effects of cannabis on driving.

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