Stability- Indicating Novel HPLC Method for Estimation of Bendamustine in Pharmaceutical Forms

A simple, precise, novel stability-indicating method was developed and validated for the estimation of Bendamustine in pharmaceutical forms. The Assay method was developed by using a detector set at 235nm and reverse phase c18 column (Inertsil ODS-2,150mmx4.6mm, 5 μm).The mobile phase consisted of 0.1%Trifluoroacetic acid: Acetonitrile 70:30 (v/v) in isocratic flow rate at 1.5mL/min for 10minutes.Sample cooler temperature and Column oven temperature were monitored in the method at 5°c and 25°c Respectively. The retention time of bendamustine was 4.8min.The proposed method was achieved and validated with respect to precision, specificity, accuracy, linearity, range, Analyte Stability, Filter variability, Robustness. The %Recovery of Bendamustine at Each level in formulations was found to be in a range of 98-102%. The linearity of the assay was established in the range of 5.053 to 75.793 µg/mL with correlation coefficient (R2) > 0.9999. The limits of detection and quantification were 0.03 and 0.05mg/mL, respectively. Specificity of the assay demonstrates no interference from and degrading products formed by alkaline, acidic, oxidative, UV light and high temperature (Thermal) conditions. This method showed more accurate, rugged and reliable determination of Bendamustine for drug stability assay in pharmaceutical studies.