A Comparative Review of Hydrophilic Interaction Chromatography(HILIC) with Traditional Chromatography in the Analysis of Aminoglycoside Antibiotics

Aminoglycoside antibiotics remain crucial for treating severe Gram-negative infections, yet their
highly polar, polycationic nature presents ongoing analytical challenges. Traditional reversed-phase
chromatography often proves inadequate, offering insufficient retention and poor peak symmetry
due to limited interactions with hydrophobic stationary phases. Consequently, analytical techniques
have increasingly shifted toward hydrophilic interaction liquid chromatography (HILIC), which
offers improved retention, enhanced selectivity, and better compatibility with mass spectrometric
detection. This review critically explores the development of chromatographic methods for aminoglycoside
quantification, emphasizing methodological advancements, practical challenges, and the
application of modern HILIC–MS/MS systems. Key structural elements of aminoglycosides, such as
multiple amino and hydroxyl groups and the presence of 2-deoxystreptamine, significantly affect
chromatographic behavior. Understanding these molecular features is essential for optimizing
mobile-phase pH, buffer strength, and organic solvent composition to achieve effective separation.
Recent research indicates that optimized HILIC methods can address previous issues related to
ion suppression, low retention, and complex sample matrices, particularly in biological, pharmaceutical,
and food safety analyses. Despite significant progress, challenges persist in method standardization,
consistency across laboratories, and the development of universally applicable
chromatographic conditions. This review compiles existing evidence, highlighting the strengths
and limitations of current analytical methods, and suggests future directions for creating more
sensitive, reproducible, and high-throughput methodologies. By integrating structural insights with
modern chromatographic advancements, the review offers a comprehensive perspective aimed at
enhancing aminoglycoside analysis across various research and regulatory settings.