The aim of this work was to develop and validate a liquid chromatography tandem mass spectrometry method for detecting of the main cannabinoids, of cannabinol (CBN) and tetrahydrocannabinol (THC) and… Click to show full abstract
The aim of this work was to develop and validate a liquid chromatography tandem mass spectrometry method for detecting of the main cannabinoids, of cannabinol (CBN) and tetrahydrocannabinol (THC) and the primary metabolite 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THC-COOH) in hair samples. Extraction of the cannabinoids was carried out by a polymeric strong anion mixed mode solid phase extraction cartridge and then employing methanolic HCl followed by 2-fluoro-1-methyl pyridinium-p-toluenesulfonate (FMP-TS) as a derivatization procedure of carboxyl and phenolic groups respectively offering enhanced sensitivity for the detection of THC-COOH in hair matrices. Formation of a methyl ester increased its lipophilicity and removed the negative charge on the carboxyl group. Calibration curves were prepared over the range of 0.02-4 pg/mg of hair for THC and CBN and 0.2-12 pg/mg of hair for THC-COOH. The extraction recovery was between 81% to 105 % for all compounds. The limit of detection (LOD) and quantification (LOQ) were 2 and 20 pg/mg respectively for both CBN and THC and 0.1 and 0.2 pg/mg respectively for THC-COOH, which met the society of hair testing recommendation. Intra- and inter-assay precision were always lower than 4 and 11% respectively for these cannabinoids. Whereas, intra- and inter-assay bias were between 14 and -18 % and 15 and -12 % respectively. Twenty-seven hair specimens from cannabis users were investigated. The concentrations of CBN, THC and THC-COOH gave ranges of (0.022-2.562 ng/mg), (0.049-0.431 ng/mg) and (0.222-4.867 pg/mg) respectively. This new method of derivatization improves the LOD to ensure detection of the metabolite.
               
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