The global public health crisis driven by opioid misuse necessitates the development of efficient analytical methods for forensic and clinical monitoring. This review critically examines recent advancements in the determination of opiates and opioids in biological matrices, covering the period from 2014 to 2025. The primary aim of research is to evaluate the alignment of these methodologies with the principles of Green Analytical Chemistry (GAC) to promote more sustainable laboratory practices. To this end, forty peer-reviewed studies were systematically assessed using four complementary sustainability metrics: the Analytical GREEnness Metric (AGREE), the Green Analytical Procedure Index (GAPI), the Blue Applicability Grade Index (BAGI), and the Red-Green-Blue (RGB) model, also known as White Analytical Chemistry. These tools provide a multidimensional perspective, integrating procedural greenness, practical applicability, and analytical performance. The results reveal a significant variability in environmental performance, with miniaturised, solvent-free, and automated techniques outperforming conventional workflows that rely on toxic solvents and intensive sample preparation. This evaluation underscores the importance of adopting green metrics to guide the selection of toxicological methods, highlighting that the transition to sustainable analytical chemistry is both feasible and essential for modern bioanalysis sustainability assessment in selecting and developing toxicological methodologies. • Critical evaluation of 40 recent studies (2014-2025) on opioid bioanalysis. • Multidimensional sustainability assessment using AGREE, GAPI, BAGI, and RGB metrics. • Sensor-based and electrochemical techniques emerged as the most sustainable analytical pathways. • Miniaturised methods (MEPS, SPME, DBS) show high but heterogeneous greenness profiles. • Integration of green metrics identifies key trade-offs between performance and eco-responsibility.
Rosendo et al. (Fri,) studied this question.