A simple, sensitive and rapid bioanalytical method for quantifying cannabidiol (CBD), 7-OH-CBD and 7-COOH-CBD in human plasma

Reliable methods for measuring plasma concentrations of cannabidiol (CBD) and its metabolites are essential for pharmacokinetic studies. Several methods have been published, some involve complex procedures. This study aimed to develop a simple yet robust method for quantitating CBD, 7-OH-CBD and 7-COOH-CBD in human plasma. Sample preparation was evaluated using various solvent systems. Analytical parameters were examined and optimized. The method was validated for sensitivity, accuracy, precision, selectivity, specificity, carryover, dilution integrity, long-term stability, freeze-thaw stability and recovery. A simple protein precipitation method using acetonitrile provided optimal sample preparation. All analytes were quantified in negative ionization mode, with complete separation achieved on a C18 column within 3.5 minutes using an isocratic mobile phase. The method met all validation criteria. Analytes remained stable under long-term storage at −30°C, through freeze-thaw cycles and at refrigerated temperatures, although 7-COOH-CBD showed a slight decrease after two months at refrigerated temperatures. This study presents a simple, rapid and robust bioanalytical method for the simultaneous quantification of CBD and its major metabolites across clinically relevant plasma concentrations. The method’s reliability and efficiency make it highly suitable for routine clinical use and pharmacokinetic applications. Cannabidiol (CBD) is a compound found in cannabis that is being studied for its potential health benefits. It is already approved as an add-on treatment for certain types of epilepsy. To improve how CBD is used in healthcare, researchers need reliable ways to measure CBD and its breakdown products, called metabolites, in the blood. In this study, researchers developed a simple and accurate method to measure CBD and its metabolites in human blood. Existing methods are sometimes complex, and this study aimed to develop a simpler method as an alternative. It uses a technique called liquid chromatography coupled to mass spectrometry, which helps separate and identify substances in blood samples. The sample preparation was straightforward, using a solvent called acetonitrile. The method was carefully tested to make sure it was accurate, consistent, and able to detect even small amounts of CBD. It also worked well under different storage conditions, including freezing and long-term refrigeration. The results showed that this method can quickly and reliably measure CBD and its breakdown products in a single test. This makes it a useful tool for clinical studies and future research on CBD treatments.