Stability-Indicating Liquid Chromatographic Method for the Determination of Bendamustine Hydrochloride in Parenterals

A stability-indicating high-performance liquid chromatographic method was developed and validated for the determination of Bendamustine hydrochloride in injectables. Reversed-phase chromatography was performed on Shimadzu Model CBM20A/20 Alite, equipped with SPD M20A prominence photodiode array detector using C18 (250 mm × 4.6 mm, 5 μm particle size) column with acetonitrile: tetra butyl ammonium hydrogen sulphate (80:20, V/V) as mobile phase at a flow rate of 0.8 mL/min. with UV detection at 233 nm. Linearity was observed in the concentration range of 1.0–200 μg/mL (R 2 = 0.999) with regression equation y = 10691 x + 98237. The limit of quantitation (LOQ) and limit of detection (LOD) were found to be 0.812 and 0.268 μg/mL respectively. The forced degradation studies were performed by using HCl, NaOH, H2O2 etc. Bendamustine hydrochloride is more sensitive towards elevated temperatures (80oC) in comparison to acidic and oxidative conditions but very much resistant towards alkaline conditions. The method was validated as per ICH guidelines. The RSD for intra-day (0.14-0.32) and inter-day (0.47-0.66) precision were found to be less than 1 %. The percentage recovery was in good agreement with the labeled amount in the pharmaceutical formulations and the method is simple, specific, precise and accurate for the determination of Bendamustine hydrochloride in pharmaceutical formulations.