Simultaneous Analysis of Δ9-THC, Δ8-THC, CBD, and CBN in Breath Aerosols Collected Using Cannabix Technologies Breath Collection Unit

Abstract A simple, accurate, and cost-effective liquid chromatography-tandem mass spectrometric (LC-MS/MS) method has been developed and validated for the simultaneous quantification of Δ9-tetrahydrocannabinol (Δ9-THC), Δ8-tetrahydrocannabinol (Δ8-THC), cannabidiol (CBD), and cannabinol (CBN) in breath aerosols collected using the Cannabix Technologies’ Breath Collection Unit (BCU). The method is suitable for use in workplace drug testing and in the investigation of driving under the influence of drugs (DUID) cases. The method was validated according to ANSI/ASB Standard 036, First Edition 2019 guidelines. A lower limit of quantification (LLOQ) of 2.5 pg/L was achieved. Calibration curves were linear (R2 0.995) and quality control (QC) accuracy between 91.3% and 100.5% with precision (%CV) less than 17.7%. No major matrix effects or drug interferences were observed. Samples were stable at room temperature and refrigerated at 2-8 °C up to 7 days. Breath samples were collected using the BCU from volunteer regular cannabis users before cannabis consumption and at multiple timepoints after smoking cannabis up to 90 minutes. The results verified the methods ability to capture and quantitate the cannabinoids in breath. The data indicated similar concentration versus time trends for THC metabolism assessed from breath to that previously reported from blood. Breath is an alternative, non-invasive sample matrix that holds promise for identifying recent cannabis use.