Conversion of cannabidiol (CBD) to Δ9-tetrahydrocannabinol (Δ9-THC) during protein precipitations prior to plasma samples analysis by chromatography – Troubles with reliable CBD quantitation when acidic precipitation agents are applied

The growing popularity of supplements containing cannabidiol (CBD), mainly CBD oils, in self-medication of humans and the increased interest in this compound in different preclinical and clinical trials stimulates the development of procedures of CBD analysis in plasma for the study of CBD pharmacology in people and animals or in establishing dose-therapeutic effect relationships of this compound. Preliminary removal of protein by its precipitation from plasma is still one of the willingly applied plasma sample preparation methods in many analytical procedures estimating plasma drug concentration, including CBD. The present paper shows that a significant amount of CBD transforms to Δ9-tetrahydrocannabinol (Δ9-THC) in a hot GC injection system when acidic precipitation agents, such as TFA, TCA, HClO₄, H₂SO₄, ZnSO₄ or CHCl₃, are used for plasma protein precipitation. The transformation degree depends on the temperature of the GC injector, the concentration of the precipitation agent and the incubation time of plasma with the precipitating agent. At the CBD plasma concentration equal to 50 ng/ml, which is approximately the mean level for patients treated for epileptic syndromes, the CBD transformation degree can exceed 20%. For a reliable estimate of CBD in blood plasma, neutral precipitation agents (e.g. ACN, MeOH, acetone) should be used when plasma deproteinization precedes GC analysis. The presented results are important not only for analysts cooperating with pharmacologists and for medicine doctors examining the activity of CBD-containing drugs in the therapeutic process, but also for forensic scientists who may erroneously find innocent people guilty of using marijuana or its preparations.