ABSTRACT
A novel molecularly imprinted solid phase extraction (MISPE) methodology followed by liquid chromatography tandem mass spectrometry (LC-MS/MS) has been developed using cylindrical shaped molecularly imprinted pills for detection of Δ(9)-tetrahydrocannabinol (THC), 11-nor-Δ(9)-tetrahydrocannabinol carboxylic acid (THC-COOH), cannabinol (CBN) and cannabidiol (CBD) in urine and oral fluid (OF). The composition of the molecular imprinted polymer (MIP) was optimized based on the screening results of a non-imprinted polymer library (NIP-library). Thus, acrylamide as functional monomer and ethylene glycol dimethacrylate as cross-linker were selected for the preparation of the MIP, using catechin as a mimic template. MISPE pills were incubated with 0.5 mL urine or OF sample for adsorption of analytes. For desorption, the pills were transferred to a vial with 2 mL of methanol:acetic acid (4:1) and sonicated for 15 min. The elution solvent was evaporated and reconstituted in methanol:formic acid (0.1%) 50:50 to inject in LC-MS/MS. The developed method was linear over the range from 1 to 500 ng mL(-1) in urine and from 0.75 to 500 ng mL(-1) in OF for all four analytes. Intra- and inter-day imprecision were <15%. Extraction recovery was 50-111%, process efficiency 15.4-54.5% and matrix effect ranged from -78.0 to -6.1%. Finally, the optimized and validated method was applied to 4 urine and 5 OF specimens. This is the first method for the determination of THC, THC-COOH, CBN and CBD in urine and OF using MISPE technology.