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Clinical Chemistry and Laboratory Medicine (CCLM)

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

Jacqueline A. HubbardORCID iD: https://orcid.org/0000-0003-4092-7153 / Breland E. Smith / Philip M. Sobolesky
  • Department of Pathology and Laboratory Medicine, Santa Clara Valley Medical Center, San Jose, CA, USA
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Sollip Kim
  • Department of Laboratory Medicine, Inje University Ilsan Paik Hospital, Ilsan Seo-gu, Goyang, Republic of Korea
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Melissa A. Hoffman / Judith Stone
  • University of California, San Francisco Medical Center, Laboratory Medicine, Parnassus Chemistry, San Francisco, CA, USA
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  • De Gruyter OnlineGoogle Scholar
/ Marilyn A. Huestis
  • The Lambert Center for the Study of Medicinal Cannabis and Hemp, Institute for Emerging Health Professions, Thomas Jefferson University, Philadelphia, PA, USA
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ David J. Grelotti / Igor Grant / Thomas S. Marcotte / Robert L. Fitzgerald
Published Online: 2019-09-17 | DOI: https://doi.org/10.1515/cclm-2019-0600



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.


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.


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.


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.

This article offers supplementary material which is provided at the end of the article.

Keywords: breath; cannabis; LC-MS/MS; mass spectrometry; THC; whole blood


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About the article

Corresponding author: Jacqueline A. Hubbard, PhD, Department of Pathology, University of California, 10300 Campus Point Drive, San Diego, CA 92121, USA, E-mail: jahubbard@ucsd.edu

Received: 2019-06-13

Accepted: 2019-08-21

Published Online: 2019-09-17

Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

Research funding: This work was supported by the State of California via the Medical Marijuana Regulation and Safety Act (Assembly Bill 266). Additional support was provided by the Center for Medicinal Cannabis Research at the University of California, San Diego, which receives support from the Control, Regulate, and Tax Adult Use of Marijuana Act (California Proposition 64) and philanthropic gifts from the Wholistic Research and Education Foundation.

Employment or leadership: None declared.

Honorarium: None declared.

Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

Citation Information: Clinical Chemistry and Laboratory Medicine (CCLM), 20190600, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/cclm-2019-0600.

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