RECENT ADVANCES IN LC–MS/MS ANALYSIS: PRINCIPLES, METHOD DEVELOPMENT, VALIDATION, AND APPLICATIONS IN PHARMACEUTICAL AND BIOMEDICAL SCIENCES

Liquid Chromatography–Tandem Mass Spectrometry (LC–MS/MS) has emerged as one of the most powerful analytical techniques in modern pharmaceutical, clinical, biomedical, environmental, and food sciences. The integration of high-performance liquid chromatography with tandem mass spectrometry combines efficient chromatographic separation with highly selective and sensitive mass detection, enabling accurate qualitative and quantitative analysis of a wide range of compounds in complex biological and environmental matrices. Over the past two decades, significant technological advancements in chromatographic systems, ionization interfaces, mass analyzers, and data-processing software have substantially enhanced the analytical performance of LC–MS/MS. These developments have resulted in improved sensitivity, lower limits of detection and quantification, higher sample throughput, and greater reproducibility.LC–MS/MS has become the preferred analytical platform for bioanalytical method development, pharmacokinetic investigations, bioequivalence studies, therapeutic drug monitoring, metabolomics, proteomics, forensic toxicology, environmental monitoring, and food safety assessment. The technique provides excellent specificity through multiple reaction monitoring (MRM), allowing simultaneous quantification of multiple analytes even at trace concentration levels. Furthermore, recent innovations including ultra-high-performance liquid chromatography (UHPLC), high-resolution mass spectrometry (HRMS), hybrid mass analyzers, microflow chromatography, automated sample preparation, and artificial intelligence-assisted data analysis have further expanded its applications in pharmaceutical research and precision medicine.Recent technological advances and future perspectives are also highlighted to provide researchers, academicians, and pharmaceutical professionals with an updated overview of this rapidly evolving analytical platform. Keywords: LC–MS/MS, Liquid Chromatography–Tandem Mass Spectrometry, Drug Analysis.