SIMULTANEOUS QUANTIFICATIONOF HUMANMONOCLONAL ANTIBODIES: RELATLIMABandNIVOLUMAB BY RP-HPLC METHOD

The aim of the current investigation is the simultaneous quantitation of Nivolumab and Relatlimab in a pure formaswell as in formulation. The chromatographic separation was achieved on Agilent (4.6 mm × 150 mm; 5µm), withacombination of Acetonitrile and ortho-phosphoric acid (0.1%) in a proportion of 45:55 v/v. Flowrate is OnemL/minute, the detection wavelength at 221 nm. Analysis of linearity within a sample strength in range of 12- 72ppm of Relatlimab and 4-24 ppm of Nivolumab with correlation coefficients of 0.9998 and 0.9999, respectively. Relatlimab and Nivolumab retention times - 2.470 min. and 3.423 min., correspondingly. Relatlimab and NivolumabLoD’s, LoQ’s were 0.08 ppm, 0.26 ppm and 0.03 ppm, 0.08 ppm, respectively. The method was validatedfor selectivity, specificity, accuracy, linearity, range, precision, quantification limit and limit of detection, androbustness. Stability indicating studies were performed by Acid, Alkali, peroxide, thermal, UV light and water.