Grafinger, KatharinaKatharinaGrafinger0000-0002-3647-7455Weinmann, WolfgangWolfgangWeinmann2024-09-222024-09-222021-03-12https://boris-portal.unibe.ch/handle/20.500.12422/45875The highest concentrated metabolite of (-)-trans-Δ9-tetrahydrocannabinol (THC) in urine, the main psychoactive constituent of Cannabis sativa, is 11-nor-9-carboxy-(-)-trans-Δ9-tetrahydrocannabinol-β-D-glucuronide [(-)-trans-THC-COOH-Gluc]. Even though reference standards for THC, 11-hydroxy-THC (11-OH-THC) and THC-COOH are commercially available as the biological (-)-trans-stereoisomers, the reference standard of THC-COOH-Gluc is only available as the racemic 11-nor-9-carboxy-(±)-cis-Δ9-tetrahydrocannabinol-β-D-glucuronide. This poses the problem for immunoassays, because different stereoisomers may have different cross-reactivity (CR). The aim of the current study was to extract the biological stereoisomer (-)-trans-THC-COOH-Gluc from a urine sample of two marihuana consumers by solid-phase extraction with a Chromabond® C18 cartridge. The cannabinoids in the obtained extract were quantified by LC-MS-MS and used after dilution for further testing of the CR of (-)-trans-THC-COOH-Gluc with a homogenous enzyme immunoassay assay (hEIA) (Urine HEIA® Cannabinoids (THC), Immunalysis™, Pomona, CA, USA). The CR was determined as the measured HEIA® signal (ng/mL) per THC-COOH-Gluc concentration (ng/mL) in percentage. Results showed that the CR (determined in concentration ratios) is concentration dependent and is 72 to 87% in the calibration range (20-50 ng/mL). At the cut-off of the hEIA (40 ng/mL) the CR was determined to be 75%. With a molecular weight quotient of 1.51 (MWTHC-COOH-Gluc/MWTHC-COOH = 520.568 g/mol/344.451 g/mol), this means that cross-reactivity (in molar ratios) is 106-131%. This finding is important, since the major metabolite of THC in urine is (-)-trans-THC-COOH-Gluc and not (-)-trans-THC-COOH, which is used for calibration and no hydrolysis is performed during the determination by hEIA.en600 - Technology::610 - Medicine & healtharticle10.7892/boris.1447843251895410.1093/jat/bkaa063