Analytical technologies for body fluids and tissues have advanced substantially over the past decade, particularly in chromatographic, electrophoretic and ion-mobility-based separation strategies designed to address matrix complexity and structural isomerism. Improvements in stationary-phase chemistry, multidimensional liquid chromatography, capillary electrophoresis and mobility-integrated mass spectrometry have enhanced peak capacity, structural discrimination, remains the selection and validation of separation strategies capable of minimizing coelution, ion suppression and quantitative variability in heterogeneous matricesremains the selection and validation of separation strategies capable of minimizing coelution, ion suppression and quantitative variability in heterogeneous matrices. This review examines recent progress in multidimensional chromatography, mobility-resolved workflows, miniaturized separation systems and hybrid imaging-separation platforms, with emphasis on comparative performance, practical implementation constraints and translational reproducibility. Applications across plasma, serum, urine, saliva, cerebrospinal fluid (CSF), tissues and spatially resolved analyses are discussed in the context of analyte chemistry and matrix-dependent method selection. Future progress will depend on standardized multidimensional workflows, validated performance metrics, harmonized reporting frameworks and data-driven optimization strategies that support robust clinical translation. By focusing on separation-centred decision-making and quality assurance, this review provides guidance for analytical scientists seeking reliable molecular characterization of complex human biospecimens.
Panigrahy et al. (Mon,) studied this question.
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