Automated Workflow for High-Throughput LC–MS/MS-Based Therapeutic Monitoring of Cannabidiol and 7-Hydroxy-cannabidiol in Patients with Epilepsy

Introduction This database includes the raw data linked with the paper “Automated Workflow for High-Throughput LC–MS/MS-Based Therapeutic Monitoring of Cannabidiol and 7-Hydroxy-cannabidiol in Patients with Epilepsy ”. This study presents the development and validation of a fully automated workflow for preparing serum samples to quantify cannabidiol (CBD) and its active metabolite, 7-hydroxy-CBD, using liquid chromatography coupled with tandem mass spectrometry (LC–MS/MS). The workflow, implemented on an automated platform, performs critical steps—including solvent dispensing, mixing, centrifugation, filtration, and supernatant transfer—resulting in 96-well plates ready for analysis. Human serum samples were collected from epilepsy patients undergoing CBD therapy and were processed using both manual and automated methods to assess agreement. Quantification was carried out via LC–MS/MS with cannabidiol-d₃ as the internal standard. Method validation, performed according to European Medicines Agency guidelines, confirmed that the automated protocol met the recommended criteria for both intraday and interday accuracy and precision. Calibration curves exhibited excellent linearity across the tested concentration ranges. Comparative analysis using Passing–Bablok regression and Bland–Altman plots revealed strong agreement between manual and automated approaches. These results support the clinical utility of the automated method for therapeutic drug monitoring (TDM) of CBD and 7-hydroxy-CBD. Its robust performance and scalability also provide a strong foundation for expanding the analytical panel to include a broader spectrum of antiseizure medications, promoting more standardized and effective TDM practices in clinical settings.