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Validated method for the simultaneous determination of Δ-THC and Δ-THC-COOH in oral fluid, urine and whole blood using solid-phase extraction and liquid chromatography–mass spectrometry with electrospray ionization
Authors:Helena Teixeira, Alain Verstraete, Paula Proen  a, Francisco Corte-Real, Paula Monsanto,Duarte Nuno Vieira
Affiliation:aDelegation of Coimbra, National Institute of Legal Medicine, Coimbra, Portugal;bFaculty of Medicine, Coimbra University, Coimbra, Portugal;cLaboratory of Clinical Biology-Toxicology, Ghent University Hospital, Ghent, Belgium
Abstract:A fully validated, sensitive and specific method for the extraction and quantification of Δ9-tetrahydrocannabinol (THC) and 11-nor-9-carboxy-Δ9-THC (THC-COOH) and for the detection of 11-hydroxy-Δ9-THC (11-OH THC) in oral fluid, urine and whole blood is presented. Solid-phase extraction and liquid chromatography–mass spectrometry (LC–MS) technique were used, with electrospray ionization. Three ions were monitored for THC and THC-COOH and two for 11-OH THC. The compounds were quantified by selected ion recording of m/z 315.31, 329.18 and 343.16 for THC, 11-OH THC and THC-COOH, respectively, and m/z 318.27 and 346.26 for the deuterated internal standards, THC-d3 and THC-COOH-d3, respectively. The method proved to be precise for THC and THC-COOH both in terms of intra-day and inter-day analysis, with intra-day coefficients of variation (CV) less than 6.3, 6.6 and 6.5% for THC in saliva, urine and blood, respectively, and 6.8 and 7.7% for THC-COOH in urine and blood, respectively. Day-to-day CVs were less than 3.5, 4.9 and 11.3% for THC in saliva, urine and blood, respectively, and 6.2 and 6.4% for THC-COOH in urine and blood, respectively. Limits of detection (LOD) were 2 ng/mL for THC in oral fluid and 0.5 ng/mL for THC and THC-COOH and 20 ng/mL for 11-OH THC, in urine and blood. Calibration curves showed a linear relationship for THC and THC-COOH in all samples (r2 > 0.999) within the range investigated.The procedure presented here has high specificity, selectivity and sensitivity. It can be regarded as an alternative method to GC–MS for the confirmation of positive immunoassay test results, and can be used as a suitable analytical tool for the quantification of THC and THC-COOH in oral fluid, urine and/or blood samples.
Keywords:Cannabinoids   Oral fluid   Urine   Blood   LC–  MS
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