NOVEL RP-HPLC AND UV METHOD DEVELOPMENT AND VALIDATION FOR INOTUZUMAB OZOGAMICIN IN BULK AND ITS PHARMACEUTICAL DOSAGE FORM

A simple, Precise, Accurate method was developed for the estimation of Inotuzumab Ozogamicin by RP-HPLC and UV technique. Chromatographic conditions used were stationary phase Discovery c18 150 × 4.6 mm, 5 m. Mobile phase O- phosphoric acid buffer: Acetonitrile in the ratio of 50v:50v and the flow rate were maintained at 1 ml/min, detection wavelength at 240nm, column temperature was set to 30 ºC, and diluents was Acetonitrile: Water (50v:50v), Conditions were finalized as an optimized method. System suitability parameters were studied by five times standard was injected, and results were well under acceptance criteria. Linearity study was carried out between 4.5 µg/ml to 27 µg/ml levels, R² value was found to be as 0.999. Precision was observed to be less than 2% for both repeatability and intermediate precision. LOD and LOQ are found to be 0.033 µg/ml and 0.099 µg/ml respectively. The explored HPLC method was fast, simple, sensitive, accurate and precise for the estimation of Inotuzumab Ozogamicin in bulk and its dosage form. The compounds were subjected to forced degradation applying several stress conditions.  The proposed method is specific and stability-indicating. Hence, the developed method can be adapted to regular quality control analysis.