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Validation of a liquid chromatography tandem mass spectrometry (LC-MS/MS) method to detect cannabinoids in whole blood and breath.
Clin Chem Lab Med. 2020 04 28; 58(5):673-681.CC

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.

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Publisher Full Text (DOI)

Authors+Show Affiliations

Hubbard JA
Department of Pathology, University of California, San Diego, CA 92121, USA.
Smith BE
Insource Diagnostics, Monrovia, CA, USA.
Sobolesky PM
Department of Pathology and Laboratory Medicine, Santa Clara Valley Medical Center, San Jose, CA, USA.
Kim S
Department of Laboratory Medicine, Inje University Ilsan Paik Hospital, Ilsan Seo-gu, Goyang, Republic of Korea.
Hoffman MA
Department of Pathology, University of California, San Diego, CA 92121, USA.
Stone J
University of California, San Francisco Medical Center, Laboratory Medicine, Parnassus Chemistry, San Francisco, CA, USA.
Huestis MA
The Lambert Center for the Study of Medicinal Cannabis and Hemp, Institute for Emerging Health Professions, Thomas Jefferson University, Philadelphia, PA, USA.
Grelotti DJ
Department of Psychiatry, University of California, San Diego, CA, USA.
Grant I
Department of Psychiatry, University of California, San Diego, CA, USA.
Marcotte TD
Department of Psychiatry, University of California, San Diego, CA, USA.
Fitzgerald RL
Department of Pathology, University of California, San Diego, CA 92121, USA.

MeSH

Breath TestsCannabidiolCannabinoidsChromatography, High Pressure LiquidCitric AcidDronabinolGlucoseHumansLimit of DetectionQuality ControlReference StandardsReproducibility of ResultsSmokingSolid Phase ExtractionTandem Mass SpectrometryValidation Studies as Topic

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31527291

Clinical Trial Links

The Impact and Detection of Driving Impairments Associated With Acute Cannabis Smoking
Effects of Cannabis/Alcohol on Driving Performance and Field Sobriety Tests

Citation

Hubbard, Jacqueline A., et al. "Validation of a Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS) Method to Detect Cannabinoids in Whole Blood and Breath." Clinical Chemistry and Laboratory Medicine, vol. 58, no. 5, 2020, pp. 673-681.
Hubbard JA, Smith BE, Sobolesky PM, et al. Validation of a liquid chromatography tandem mass spectrometry (LC-MS/MS) method to detect cannabinoids in whole blood and breath. Clin Chem Lab Med. 2020;58(5):673-681.
Hubbard, J. A., Smith, B. E., Sobolesky, P. M., Kim, S., Hoffman, M. A., Stone, J., Huestis, M. A., Grelotti, D. J., Grant, I., Marcotte, T. D., & Fitzgerald, R. L. (2020). Validation of a liquid chromatography tandem mass spectrometry (LC-MS/MS) method to detect cannabinoids in whole blood and breath. Clinical Chemistry and Laboratory Medicine, 58(5), 673-681. https://doi.org/10.1515/cclm-2019-0600
Hubbard JA, et al. Validation of a Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS) Method to Detect Cannabinoids in Whole Blood and Breath. Clin Chem Lab Med. 2020 04 28;58(5):673-681. PubMed PMID: 31527291.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Validation of a liquid chromatography tandem mass spectrometry (LC-MS/MS) method to detect cannabinoids in whole blood and breath. AU - Hubbard,Jacqueline A, AU - Smith,Breland E, AU - Sobolesky,Philip M, AU - Kim,Sollip, AU - Hoffman,Melissa A, AU - Stone,Judith, AU - Huestis,Marilyn A, AU - Grelotti,David J, AU - Grant,Igor, AU - Marcotte,Thomas D, AU - Fitzgerald,Robert L, PY - 2019/06/13/received PY - 2019/08/21/accepted PY - 2019/9/19/pubmed PY - 2021/5/21/medline PY - 2019/9/19/entrez KW - LC-MS/MS KW - THC KW - breath KW - cannabis KW - mass spectrometry KW - whole blood SP - 673 EP - 681 JF - Clinical chemistry and laboratory medicine JO - Clin Chem Lab Med VL - 58 IS - 5 N2 - 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. SN - 1437-4331 UR - https://www.unboundmedicine.com/medline/citation/31527291/Validation_of_a_liquid_chromatography_tandem_mass_spectrometry__LC_MS/MS__method_to_detect_cannabinoids_in_whole_blood_and_breath_ DB - PRIME DP - Unbound Medicine ER -