Design and Verification of A opposed Polarity Separation Technique for Zenocutuzumab in Large-quantity Drug Substance and Delivery System Forms

Abstract

Background: Zenocutuzumab is a novel therapeutic monoclonal antibody requiring precise and reliable investigative techniques for routine monitoring, quality, and stability evaluation. The present investigation is
directed to progress and confirm the perceptive a rapid, sensitive, and robust opposing polarity chromatographic separation method for the quantitative assessment of Zenocutuzumab in majority medicine substance and pharmaceutical therapeutic preparations. Materials and Methods: Effective separation of components was obtained by employing are versed-phase an reversed-phase C18 chromatographic column analytical column (150 mm × 4.6 mm, 3.5 µm particle diameter) served as the separation medium under isocratic conditions. The eluent consisted of KH2 PO4  Buffer solution titrated to pH 2.5 with orthophosphoric acid) and ethanol in the volumetric ratio of 70:30 volume-to-volume. A constant flow rate of 1.0 mL/min was maintained throughout, and analysis was performed at ambient temperature. The analytical procedure was validated in accordance with the requirements of International Council for Harmonization (ICH) guidelines. Stability-indicating stress studies were conducted. The sample was subjected to acidic, basic, and oxidative environments, reductive, photolytic, hydrolytic, and thermal stress conditions. Results: Zenocutuzumab showed a linear response spanning the concentration range of 25–150 µg/mL. The detection and quantification limits were evaluated to be 0.6 µg/mL and 2.0 µg/mL, respectively. The total run time was <5 min. Precision and accuracy studies yielded results within acceptable limits. Validation parameters confirmed the method’s specificity, linearity, robustness, and reproducibility. The method successfully quantified Zenocutuzumab in both bulk and formulated samples and effectively separated degradation products during stability studies. Conclusion: The established reversed-phase
high-performance liquid chromatography method is easy to perform, quick, and precise, and reliable for the quantitative determination of Zenocutuzumab. Its compliance with ICH validation criteria and ability to assess stability under various stress conditions make it suitable for standard quality control and stability monitoring testing of Zenocutuzumab in drug formulation systems.