Abstract

DEVELOPMENT AND VALIDATION OF A STABILITY-INDICATING RP-HPLC METHOD FOR THE DETERMINATION OF BENDAMUSTINE IN RAW MATERIAL AND FINISHED PRODUCT

Dhananjaya. G*, M. Naveen, P. Janaki Pathi, J. Raghuram and N. Appala Raju

Abstract

Bendamustine Hydrochloride Injection is indicated for the treatment of patients with chronic lymphocytic leukemia. Efficacy relative to first line therapies other than chlorambucil has not been established. Bendamustine Hydrochloride Injection is indicated for the treatment of patients with indolent B-cell non-Hodgkin lymphoma that has progressed during or within six months of treatment with rituximab or a rituximab-containing regimen. A reversed-phase HPLC method was developed and validated for the determination of Bendamustine in raw material and to determine impurities and degradants that may developed in the tested samples. The separation was achieved on Thermo Hypersil C18 column (4.6 x 250 mm, 5μm) using mobile phase consisting of 70% Potassium Phosphate Dibasic Buffer (pH 7) and 30% acetonitrile (100% ACN). The flow rate was 1.0 mL/min, injection volume 10μL, and detection was accomplished at 232 nm. The retention time for Bendamustine was 7 minutes. The developed method was validated and met all the acceptance criteria for validation parameters - system suitability, specificity, solution stability, robustness, linearity, accuracy, precision, limit of detection (LOD), and limit of quantitation (LOQ). The LOD was determined to be 0.1 ppm and LOQ was found to be 0.5 ppm.

Keywords: Bendamustine, Degradation, RP-HPLC, Validation and Finished product.