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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. Hubbard ORCID logo EMAIL logo , Breland E. Smith , Philip M. Sobolesky , Sollip Kim , Melissa A. Hoffman , Judith Stone , Marilyn A. Huestis , David J. Grelotti , Igor Grant , Thomas D. Marcotte and Robert L. Fitzgerald
Published/Copyright: September 17, 2019
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Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 58 Issue 5

Abstract

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.

Keywords: breath; cannabis; LC-MS/MS; mass spectrometry; THC; whole blood
Corresponding author: Jacqueline A. Hubbard, PhD, Department of Pathology, University of California, 10300 Campus Point Drive, San Diego, CA 92121, USA, E-mail:

Acknowledgments

The authors would like to thank Waters Corporation for their support, training, and technical assistance throughout this project.

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

  2. 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.

  3. Employment or leadership: None declared.

  4. Honorarium: None declared.

  5. 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.

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Supplementary Material

The online version of this article offers supplementary material (https://doi.org/10.1515/cclm-2019-0600).

Received: 2019-06-13
Accepted: 2019-08-21
Published Online: 2019-09-17
Published in Print: 2020-04-28

©2020 Walter de Gruyter GmbH, Berlin/Boston

Articles in the same Issue

  1. Frontmatter
  2. Editorial
  3. Advancements in mass spectrometry as a tool for clinical analysis: Part I
  4. Drug adherence, testing and therapeutic monitoring
  5. Hyphenated mass spectrometry techniques for assessing medication adherence: advantages, challenges, clinical applications and future perspectives
  6. Method development for quantitative determination of seven statins including four active metabolites by means of high-resolution tandem mass spectrometry applicable for adherence testing and therapeutic drug monitoring
  7. Validation of a liquid chromatography tandem mass spectrometry (LC-MS/MS) method to detect cannabinoids in whole blood and breath
  8. THC and CBD concentrations in blood, oral fluid and urine following a single and repeated administration of “light cannabis”
  9. Identification of metabolites of peptide-derived drugs using an isotope-labeled reporter ion screening strategy
  10. Validation according to European and American regulatory agencies guidelines of an LC-MS/MS method for the quantification of free and total ropivacaine in human plasma
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  17. Validation of a rapid, comprehensive and clinically relevant amino acid profile by underivatised liquid chromatography tandem mass spectrometry
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  20. Serum bile acids profiling by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and its application on pediatric liver and intestinal diseases
  21. LC-MS/MS analysis of plasma glucosylsphingosine as a biomarker for diagnosis and follow-up monitoring in Gaucher disease in the Spanish population
  22. Dried blood spots and alternative sample mediums
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https://doi.org/10.1515/cclm-2019-0600
Keywords for this article
breath; cannabis; LC-MS/MS; mass spectrometry; THC; whole blood

Articles in the same Issue

  1. Frontmatter
  2. Editorial
  3. Advancements in mass spectrometry as a tool for clinical analysis: Part I
  4. Drug adherence, testing and therapeutic monitoring
  5. Hyphenated mass spectrometry techniques for assessing medication adherence: advantages, challenges, clinical applications and future perspectives
  6. Method development for quantitative determination of seven statins including four active metabolites by means of high-resolution tandem mass spectrometry applicable for adherence testing and therapeutic drug monitoring
  7. Validation of a liquid chromatography tandem mass spectrometry (LC-MS/MS) method to detect cannabinoids in whole blood and breath
  8. THC and CBD concentrations in blood, oral fluid and urine following a single and repeated administration of “light cannabis”
  9. Identification of metabolites of peptide-derived drugs using an isotope-labeled reporter ion screening strategy
  10. Validation according to European and American regulatory agencies guidelines of an LC-MS/MS method for the quantification of free and total ropivacaine in human plasma
  11. Enhanced specificity due to method specific limits for relative ion intensities in a high-performance liquid chromatography – tandem mass spectrometry method for iohexol in human serum
  12. Small molecule biomarkers
  13. Applying mass spectrometry-based assays to explore gut microbial metabolism and associations with disease
  14. Trimethylamine-N-oxide (TMAO) determined by LC-MS/MS: distribution and correlates in the population-based PopGen cohort
  15. Development of a total serum testosterone, androstenedione, 17-hydroxyprogesterone, 11β-hydroxyandrostenedione and 11-ketotestosterone LC-MS/MS assay and its application to evaluate pre-analytical sample stability
  16. Short-term stability of free metanephrines in plasma and whole blood
  17. Validation of a rapid, comprehensive and clinically relevant amino acid profile by underivatised liquid chromatography tandem mass spectrometry
  18. UPLC-MS/MS method for determination of retinol and α-tocopherol in serum using a simple sample pretreatment and UniSpray as ionization technique to reduce matrix effects
  19. Independent association of plasma xanthine oxidoreductase activity with serum uric acid level based on stable isotope-labeled xanthine and liquid chromatography/triple quadrupole mass spectrometry: MedCity21 health examination registry
  20. Serum bile acids profiling by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and its application on pediatric liver and intestinal diseases
  21. LC-MS/MS analysis of plasma glucosylsphingosine as a biomarker for diagnosis and follow-up monitoring in Gaucher disease in the Spanish population
  22. Dried blood spots and alternative sample mediums
  23. Investigating the suitability of high-resolution mass spectrometry for newborn screening: identification of hemoglobinopathies and β-thalassemias in dried blood spots
  24. Candidate reference method for determination of vitamin D from dried blood spot samples
  25. Therapeutic drug monitoring of anti-epileptic drugs – a clinical verification of volumetric absorptive micro sampling
  26. Simultaneous quantitation of five triazole anti-fungal agents by paper spray-mass spectrometry
  27. Obtaining information from the brain in a non-invasive way: determination of iron in nasal exudate to differentiate hemorrhagic and ischemic strokes
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