Determination of enzymatic hydrolysis efficiency in detection of cannabis use by ultra performance liquid chromatography by tandem mass spectrometry method

Marijuana is still the most widely used illegal drug in the world. Selective, rapid and reliable methods are needed to identify cannabis use in biological fluids. Urine is the most preferred biological material because it can be detected for a long time in the detection of cannabis use. In these analyzes of urine, enzymatic hydrololysis is applied. In this study, it was aimed to determine the enzymatic hydrolysis efficiency in the detection of cannabis use by ultra-performance liquid chromatography tandem mass spectrometry method. In this thesis study, enzyme concentration, pH and temperature parameters were investigated to determine the efficiency of enzymatic hydrolysis. Unlike the literature, the enzymatic hydrolysis efficiency was investigated experimentally at temperatures of 75 °C and 90 °C. After the enzymatic hydrolysis optimization, the developed method was validated according to the parameters of the European Medicines Agency Bioanalytical Method Guide and analysis of real samples was performed. It was determined that the enzymatic hydrolysis efficiency was higher at 75°C and 90°C compared to the literature. Since similar results were obtained for these two temperatures, 75°C was chosen for the validated method. The calibration curve range was linearly determined from 1-2000 ng/mL. The method, which was validated according to the European Medicines Agency's Bioanalytical Method Guide, was successfully applied to real sample analyzes and total 11-nor-9-carboxy-Δ9-THC concentrations were determined. The results showed that this developed method can be used quickly, easily and safely in determining cannabis use.