title: Molecularly imprinted solid phase extraction for simultaneous determination of Δ9-tetrahydrocannabinol and its main metabolites by gas chromatography-mass spectrometry in urine samples.
abstract: In presented paper analytical method based on solid-phase extraction using molecularly imprinted polymer and gas chromatography-mass spectrometry has been developed and validated for the confirmation of THC, THC-OH and THC-COOH in urine samples. Non-covalent molecularly imprinted polymers of THC-OH were prepared using different functional monomers (methacrylic acid, 4-vinylpyridine, and 2-hydroxyethyl methacrylate), ethylene glycol dimethacrylate as a cross-linker and 2,2'-azobis-isobutyronitrile as an initiator of radical polymerization. Analytes were extracted from urine samples using prepared polymer sorbent with highest binding selectivity and capability. Before extraction, urine samples were hydrolyzed with alkaline. Elution was performed with chloroform:ethyl acetate (60:40, v/v). Dry extracts were silylated with BSTFA+1% TMCS. Detection and quantification were performed using gas chromatography-mass spectrometry in single ion recording mode. The developed method was linear over the range from LOQ to 150 ng mL(-1) for all three analytes. For THC, THC-OH and THC-COOH LOD was 2.5, 1 and 1 ng mL(-1), and LOQ was 3, 2 and 2 ng mL(-1), respectively. The precision, accuracy, recovery and matrix effect were investigated at 5, 25 and 50 ng mL(-1). In the investigated concentration range recoveries were 71.9% for THC, 78.6% for THC-OH and 75.2% for THC-COOH. Matrix effect was not significant (<10%) for all analytes in the concentration range from 5 ng mL(-1) to 50 ng mL(-1). Extraction recovery on non-imprinted polymer was relatively high indicating high non-specific binding. Optimized and validated method was applied to 15 post-mortem urine samples.
authors.affiliation: Department of Forensic Medicine and Criminology, School of Medicine, University of Zagreb, Šalata 11, 10000 Zagreb, Croatia. mnestic@mef.hr
authors.forename: Marina
authors.initials: M
authors.lastname: Nestić
authors.forename: Sandra
authors.initials: S
authors.lastname: Babić
authors.forename: Dragana Mutavdžić
authors.initials: DM
authors.lastname: Pavlović
authors.forename: Davorka
authors.initials: D
authors.lastname: Sutlović
mesh_terms: D013759:Dronabinol / Q000031:analogs & derivatives / Q000652:urine*; D053593:Forensic Toxicology / Q000379:methods; D008401:Gas Chromatography-Mass Spectrometry; D006213:Hallucinogens / Q000652:urine; D006801:Humans; D057230:Limit of Detection; D054802:Molecular Imprinting; D052616:Solid Phase Extraction / Q000379:methods*
chemical_list: D006213:Hallucinogens; C001427:11-hydroxy-delta(9)-tetrahydrocannabinol; C016780:11-nor-delta(9)-tetrahydrocannabinol-9-carboxylic acid; D013759:Dronabinol
keywords: Biological sample; Cannabis; Gas chromatography–mass spectrometry; Molecularly imprinted solid phase extraction