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• Analytical flexibility is crucial to track fast-evolving nitazene analogues. • Wastewater analysis enables early warning and public health surveillance. • High-resolution MS empowers putative identification of nitazene analogues. • LC–MS/MS remains the benchmark for quantitative assays in biofluids. • Miniaturised/dried sampling formats improve practicality and stability. Nitazenes, a class of high-potency novel synthetic opioids containing a benzimidazole moiety, are currently being reported across seized materials and drug-checking submissions, human biological matrices (postmortem and clinical/forensic) and community-level samples including wastewater. The analytical literature converges on a layered, matrix-oriented toolkit. Rapid presumptive strategies (vibrational fingerprints and ambient-ionization techniques) support triage in solids and field-adjacent contexts, while chromatographic separations coupled to mass spectrometry (HPLC-MS/MS/HRMS and less frequently GC–MS/MS/HRMS) deliver selectivity and structure-aware confirmation for closely related analogues. In human biofluids, both targeted triple-quadrupole panels and HRMS suspect screening workflows are present for identification; validated quantitative methods consistently address calibration model and range, sensitivity (LOD/LLOQ), precision and accuracy, selectivity, carry-over and matrix effects, often with explicit attention to isomer resolution. Cross-cutting issues recurring in the field include reference standard availability, isomeric/isobaric interferences, immunoassay cross-reactivity and the promise of miniaturised sampling formats. Organised by matrix and analytical objective, this review provides enough information to enable efficient extraction of details at the method level, while ensuring a comprehensive coverage of the most important topics.
Mandrioli et al. (Mon,) studied this question.