Development and validation of a green spectrofluorimetric method for brivaracetam determination using N-doped graphene quantum dots: Mechanistic insights and bioanalytical applications

Brivaracetam is an important antiepileptic drug, and its sensitive and selective determination in biological matrices is crucial for therapeutic drug monitoring and pharmacokinetic studies. However, the currently available analytical methods suffer from limitations such as low sensitivity, high environmental impact, and complex sample preparation. In this work, a highly sensitive and selective spectrofluorimetric method was developed for the determination of brivaracetam in human plasma and pharmaceutical formulations. The method is based on the interaction between brivaracetam and N-doped graphene quantum dots (GQDs) leading to fluorescence quenching. The quenching mechanism was investigated using density functional theory calculations, Stern-Volmer analysis and thermodynamic studies. A static quenching process was observed indicating complex formation between the analyte and nanomaterial. Different factors affecting the quenching efficiency were carefully optimized such as pH, GQDs concentration, and incubation time. The proposed method was validated according to the ICH M10 guidelines and showed excellent linearity in the concentration range of 0.1–2.5 μg/mL with a limit of detection of 0.033 μg/mL. Furthermore, the method displayed good precision and accuracy, and selectivity in the presence of common plasma/formulation excipients. Hence, the developed method was successfully applied for the determination of brivaracetam in pharmaceutical formulations as well as pharmacokinetic monitoring in human plasma samples. The “greenness”, “blueness” and “whiteness” of the proposed method was also evaluated using the AGREE, BAGI and RGB12 metrics, respectively, confirming its eco-friendly, high practicability and sustainable nature transcending the reported conventional analytical techniques, posing the developed method as a promising analytical tool for brivaracetam determination.