Analysis of Denosumab by a Validated CZE Method and Determination of Sialic Acids by the RP-HPLC Method.

This study focuses on analysis of denosumab by a validated cze method and determination of sialic acids by the rp-hplc method.. The research employs high-performance liquid chromatography (HPLC) techniques to address analytical challenges in the pharmaceutical field. A capillary zone electrophoresis (CZE) method was developed and validated to quantitate the monoclonal antibody denosumab (DmAb) and its charge variants in pharmaceutical products, demonstrating excellent precision, linearity and accuracy. Separations were obtained with migration times of 11.3 min for DmAb and the calibration curve was linear in the range... Research Background and Significance Denosumab is a monoclonal antibody widely used in the treatment of osteoporosis and certain cancers due to its role in inhibiting bone resorption. Given its therapeutic importance, accurate and reliable analytical techniques are essential for its characterization in pharmaceutical formulations. Charge variants and post-translational modifications such as sialylation can significantly affect the efficacy and safety of monoclonal antibodies like denosumab, necessitating robust analytical methods. This study addresses these needs by developing and validating a capillary zone electrophoresis (CZE) method specifically for quantifying denosumab and its charge variants. Additionally, the research integrates reversed-phase high-performance liquid chromatography (RP-HPLC) for the determination of sialic acids, which are critical glycan components influencing antibody stability and function. Together, these techniques provide a comprehensive analytical platform to support quality control and formulation assessment in pharmaceutical development. Experimental Design and Methodology The study employs two complementary analytical