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.