Hydrolytic Degradation Kinetic Study of Balofloxacin by Stability Indicating Reversed Phase High Performance Liquid Chromatography Method

Abstract

Background: Balofloxacin is a third generation fluoroquinolone with a broad antibacterial spectrum ranging from gram-positive bacteria to gram-negative bacteria. It is used in treatment of uncomplicated urinary tract infections. No stability indicating analytical method has been reported for BFX. Also stress degradation studies of Balofloxacin were not found in literature. Objective: To develop and validate a stability indicating RP-HPLC method for estimation of Balofloxacin in presence of its hydrolytic degra­dation products. Materials and Method: The chromatographic separation was performed using C 18 , Grace Smart column (250 x 4.6 mm), 5 μm as the stationary phase and Water: Acetonitrile: Tri ethylamine (72:28:1 v/v/v), pH adjusted to 3.0 using ortho-phosphoric acid as mobile phase with detec­tion wavelength 294 nm. The developed method was validated according to ICH Q2R1 guideline. Balofloxacin was subjected to degradation in acidic, alkaline and neutral conditions. Results and Discussion: The linearity was established over concentration range of 20-100 μg/ml with correlation coefficient r 2 = 0.9979. Recovery of drug was achieved in the range of 99.19–101.65%. Limit of Detection and Limit of Quantitation was found to be 4.13 and 12.51 μg/ml. Balofloxacin was found to be stable under alkaline and neutral conditions, while it degraded under acidic hydrolytic condition. The retention times for Balofloxacin and its acid degradation product were found to be 7.0 ± 0.1 and 5.7 ± 0.1 minutes, respectively. Application: The developed RP-HPLC method was applied for estimation of Balofloxacin in its tablet dosage forms and results were found to be in good agreement with the labeled claim. The method was also applied for degradation kinetic study of Balofloxacin in acidic medium. Conclusion: The developed RP-HPLC method was found to be accurate, precise, specific and sensitive. It can be applied for routine analysis (assay) of tablets containing Balofloxa­cin. The degradation of Balofloxacin in all conditions was found to be first order and highest degradation was found in 2.0 N HCl at 75°C. Key words: Stability indicating HPLC method, Balofloxacin (BFX), Degradation products, Degradation kinetic study, Design expert software–9.