title: Significant enhancement of 11-Hydroxy-THC detection by formation of picolinic acid esters and application of liquid chromatography/multi stage mass spectrometry (LC-MS(3) ): Application to hair and oral fluid analysis.
abstract: Formation of picolinic acid esters of hydroxylated drugs or their biotransformation products is a promising tool to improve their mass spectrometric ionization efficiency, alter their fragmentation behaviour and enhance sensitivity and specificity of their detection. The procedure was optimized and tested for the detection of cannabinoids, which proved to be most challenging when dealing with alternative specimens, for example hair and oral fluid. In particular, the detection of the THC metabolites hydroxyl-THC and carboxy-THC requires ultimate sensitivity because of their poor incorporation into hair or saliva. Both biotransformation products are widely accepted as incorporation markers to distinguish drug consumption from passive contamination. The derivatization procedure was carried out by adding a mixture of picolinic acid, 4-(dimethylamino)pyridine and 2-methyl-6-nitrobenzoic anhydride in tetrahydrofuran/triethylamine to the dry extraction residues. Resulting derivatives were found to be very stable and could be reconstituted in aqueous or organic buffers and subsequently analyzed by liquid chromatography-mass spectrometry (LC-MS). Owing to the complex consecutive fragmentation patterns, the application of multistage MS3 proved to be extremely useful for a sensitive identification of doubly picolinated hydroxy-THC in complex matrices. The detection limits - estimated by comparison of corresponding signal-to-noise ratios - increased by a factor of 100 following picolination. All other species examined, like cannabinol, THC, cannabidiol, and carboxy-THC, could also be derivatized exhibiting only moderate sensitivity improvements. The assay was systematically tested using hair samples and exemplarily applied to oral fluid. Concentrations of OH-THC identified in THC-positive hair samples ranged from 0.02 to 0.29pg/mg.
authors.affiliation: Institute of Doping Analysis, Dresdner str. 12, 01731, Kreischa, Germany.
authors.forename: Detlef
authors.initials: D
authors.lastname: Thieme
authors.affiliation: Albert-Ludwigs-University, Albertstr. 21, 79104, Freiburg, Germany.
authors.forename: Ulf
authors.initials: U
authors.lastname: Sachs
authors.affiliation: Forensic Toxicological Center, Bayerstr. 53, 80335, Munich, Germany.
authors.forename: Hans
authors.initials: H
authors.lastname: Sachs
authors.affiliation: Immunalysis Co, 829 Towne Center Dr, Pomonam, CA, 91767, USA.
authors.forename: Christine
authors.initials: C
authors.lastname: Moore
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods / Q000592:standards; D013759:Dronabinol / Q000032:analysis* / Q000378:metabolism; D004952:Esters; D006197:Hair / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D010848:Picolinic Acids / Q000737:chemistry* / Q000378:metabolism; D012463:Saliva / Q000737:chemistry* / Q000378:metabolism; D015813:Substance Abuse Detection / Q000379:methods / Q000592:standards*; D053719:Tandem Mass Spectrometry / Q000379:methods / Q000592:standards*
chemical_list: D004952:Esters; D010848:Picolinic Acids; D013759:Dronabinol; C030614:picolinic acid
keywords: alternative specimens; cannabinoids; derivatization; hair analysis; liquid chromatography-mass spectrometry; oral fluid