Validation of a high-performance thin-layer chromatographic method, with densitometric detection, for quantitative analysis of nebivolol hydrochloride in tablet formulations

Abstract

A quantitative densitometric high-performance thin-layer chromatographic method for determination of nebivolol hydrochloride in pharmaceutical preparations has been established and validated. Nebivolol hydrochloride from the formulations was separated and identified on silica gel 60 F 254 HPTLC plates with toluene-ethyl acetate-methanol-formic acid, 8 + 6 + 4 + 1 ( v / v ), as mobile phase. The plates were developed to a distance of 8 cm. Densitometric quantification was performed at λ = 285 nm by reflectance scanning. Well resolved bands were obtained for nebivolol hydrochloride. The method was validated for precision, recovery, robustness, specificity, and ruggedness. The calibration plot for nebivolol hydrochloride standard was linear with r = 0.9991, slope = 3.291, and intercept = 864.50. The limits of detection and quantification were 18.65 and 62.18 ng per band, respectively. The method is selective and specific with potential application in pharmaceutical analysis.