Validation of the Quantitative Determination of Tetrahydrocannabinol and Its Two Major Metabolites in Plasma by Ultra-High-Performance Liquid Chromatography–Tandem Mass Spectrometry According to the Total Error Approach*

In Belgium, driving under the influence (DUI) of cannabis is prohibited and has severe legal consequences for the driver if the blood plasma concentration of Δ(9)-tetrahydrocannabinol (THC) exceeds 1 µg/L. A method to quantify low concentrations of THC and its hydroxylated (THC-OH) and carboxylated (THC-COOH) metabolites in plasma was developed for DUI but also for other applications. Ultra-high-performance liquid chromatography coupled to mass spectrometry seems to be a very convenient method to combine fast chromatographic separation and good sensitivity. The method was validated according to total error approach. Chromatographic separation was achieved in a 3-min total run time. The limits of quantitation were lower or equal to 1 µg/L for all compounds. The linearity of the method was acceptable in the validated range of concentrations (from 0.5 to 50 µg/L for THC, from 0.9 to 50 µg/L for THC-OH and from 1.1 to 100 µg/L for THC-COOH). The biases were lower than 13%, and the relative standard deviations for repeatability and intermediate precision did not exceed 15%. Lower and upper β-expectation tolerance limits did not exceed the acceptance limits of 20% for concentrations higher than 2 µg/L for THC and THC-OH and higher than 4 µg/L for THC-COOH. The acceptance limits were 30% for THC and THC-OH concentrations lower than 2 µg/L and for THC-COOH concentrations lower than 4 µg/L.