Analytical and Bioanalytical Advances in Oxazolidinones: A Critical Review of Methods, Stability, and Clinical Translation

Oxazolidinones remain pivotal in treating multidrug-resistant infections, yet analytical and bioanalytical methodologies supporting their development and clinical use remain uneven across the class. This critical review consolidates recent advances spanning formulation quality control to therapeutic drug monitoring (TDM), offering an integrated cross-platform comparison of how analytical and bioanalytical methodologies shape oxazolidinone readiness for precision antimicrobial therapy. Analytical methods—from spectrophotometry and HPLC to LC–MS/MS—have progressed toward stability-indicating workflows, impurity profiling, and regulatory alignment, with emerging but inconsistent adoption of Quality by Design (QbD) and Green Analytical Chemistry (GAC). Bioanalytical approaches, dominated by LC–MS/MS with evolving micro-sampling and matrix-adapted extractions, have enabled pharmacokinetic characterization, exposure–response evaluation, and patient-specific dose optimization. Together, these methodologies reveal a functional divide: analytical assays secure drug identity and formulation robustness, whereas bioanalytical tools translate oxazolidinones into clinical decision systems. However, major gaps persist, including limited characterization of newer investigational analogues, poor harmonization across matrices and laboratories, and underutilization of computational, mechanistic, and eco-sustainable strategies. Bridging these gaps requires integrated, standardized, and patient-centered analytical frameworks to ensure that both established and emerging oxazolidinones remain effective in the era of escalating antimicrobial resistance.