Abstract

DETERMINATION OF ENANTIOMERIC IMPURITY IN BESIFLOXACIN HYDROCHLORIDE BY CHIRAL HIGHPERFORMANCE LIQUID CHROMATOGRAPHY

Dr. Sayyed Hussain, Dr. Amit Gosar and Tabrez Shaikh*

ABSTRACT

A sensitive simple rapid and robust new chiral normal phase HPLC method is developed for determination of Enantiomeric impurity (Impurity-A) in Besifloxacin Hydrochloride by using amylose tris (3,5- dichlorophenylcarbamate) as stationary phase with column dimension of 4.6 mm I.D × 250 mm length and 5 μm particle size (Chiralpak IE). Mobile phase consists of a mixture of Methylene dichloride, Hexane, Isopropyl alcohol, diethylamine and trifluoroacetic acid in the ratio (65:25:10:0.2:0.2). Isocratic elution mode is used for the separation using a flow rate of 0.8 mL/min and column oven temperature kept at 25°C. Detection is carried out at wavelength 295 nm. Developed method is validated as per ICH guideline and found out to be linear, accurate, specific, selective, precise, and robust. Test solution was found to be stable for 24 hrs. The method can be successfully applied for the determination of enantiomeric impurity in Besifloxacin Hydrochloride for routine and stability samples.

Keywords: Besifloxacin, Enantiomer, HPLC, method development, validation.