Development and Validation of an HPLC-Based Analytical Method for Simultaneous Quantification of Biomarkers in Human Biological Samples

Abstract

Concurrently measuring multiple biomarkers in human biological samples is crucial for clinical diagnosis, disease monitoring, and therapeutic interventions based on biomarkers. The analytical platform high-performance liquid chromatography (HPLC) has proven to be useful because of its high sensitivity, selectivity, reproducibility and ability to analyze complex biological matrices. In this study, a methodology has been developed and validated for simultaneous analysis of selected biomarkers in human serum, plasma and urine by HPLC. Optimisation of chromatographic parameters, such as selection of stationary phase, mobile phase composition, flow rate and detection conditions was performed to ensure efficient separation and accurate quantification of the target analytes. Optimized sample preparation procedures to reduce interferences and analyte recovery. The developed method was validated according to the analytical guidelines, which includes specificity, linearity, accuracy, precision, sensitivity, recovery and stability. The validation results showed excellent chromatographic performance, broad linear dynamic range, low limits of detection and quantification, as well as acceptable recoveries in various biological matrices. Overall, the proposed HPLC method offers a reliable, efficient, and scalable solution for simultaneous biomarker analysis, which can be applied to various areas such as clinical diagnostics, disease monitoring, pharmacokinetic research, and precision medicine. The results demonstrate the applicability of HPLC to the routine analysis of biomarkers in both biomedical and translational research.