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

Abstract

BackgroundThe 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.MethodsA 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.ResultsLower 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.ConclusionsThe 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.

Cannabidiol reduces withdrawal symptoms in nicotine-dependent rats

“…Blood was allowed to clot for 30 minutes at room temperature spun at 13,000 RPM for 10 minutes, and serum (100 L) was collected for processing. Nicotine, cotinine, and CBD levels were analyzed by tandem liquid chromatography-mass spectrometry as previously described (Sobolesky, Smith et al 2019, Hubbard, Smith et al 2020. Briefly, nicotine and cotinine were extracted from sera (20 L) with acetonitrile (60 L), containing 100 ng/ml D3nicotine and 300 ng/ml D3-cotinine internal standards.…”

Online Real-Time Monitoring of Exhaled Breath Particles Reveals Unnoticed Transport of Nonvolatile Drugs from Blood to Breath

“…Recently, dozens of nonvolatile drugs were observed in exhaled breath particles (EBPs) through offline particle collection followed by liquid chromatography–mass spectrometry (LC–MS) analysis. − It delivers us an important message that nonvolatile drugs can be transported via exhaled particles. Thus, sensitive and robust methods for detecting compounds in EBPs can facilitate the monitoring of nonvolatile drugs in breath air.…”

Automation System for the Flexible Sample Preparation for Quantification of Δ9-THC-D3, THC-OH and THC-COOH from Serum, Saliva and Urine

“…Limits of detection and quantification for serum sample preparation methods using LC-MS/MS ranges from 0.05-1.8 ng/mL for ∆9-THC, 0.18-3.2 ng/mL for THC-OH, and 0.05-2.8 ng/mL for THC-COOH [8,22,24,[26][27][28][29][30]. The limit of detection and quantification determined by Andersen et al for ∆9-THC is 0.2 ng/mL and 0.5 ng/mL using LC-MS, which is within the limits determined using LC-MS/MS [21].…”

“…For the purification of serum, plasma, and whole blood samples for the quantification of THC and metabolites, the most commonly used methods are protein precipitation (PPT) [22], solid-phase extraction (SPE) [24], the combination of PPT + SPE [8,21,27,29,32], liquid-liquid extraction (LLE) [25,28,30,31] and online solid-phase extraction [26] are used. Sample preparation can be performed manually [8,27,29,33], partially automated or fully automated.…”

“…Quantification of THC and metabolites is currently predominantly performed by liquid chromatography mass spectrometry (LC-MS) [21] or liquid chromatography tandem mass spectrometry (LC-MS/MS) [22][23][24][25][26][27][28][29][30], as analysis times are short and no derivatization is required compared to the previously used gas chromatography mass spectrometry (GC-MS) [5] or gas chromatography tandem mass spectrometry (GC-MS/MS) [31][32][33].…”