Abstract

Two simple, specific, accurate and precise methods, namely, reverse phase high performance liquid chromatography and high performance thin layer chromatography were developed for estimation of nebivolol hydrochloride in tablet dosage form. For the HPLC method, Lichrospher 100 C-18, 5 µm column consisting of 200×4.6 mm i.d. in isocratic mode, with mobile phase containing 50 mM KH ₂ PO ₄ buffer (pH 3.0±0.1):acetonitrile: (45:55 v/v) was used. The flow rate was 1.0 ml/min and effluent was monitored at 282 nm. The retention time was found to be 3.76±0.02 min. For the high performance thin layer chromatographic method a Camag system comprising of Linnomat V automatic sample applicator, Hamilton syringe, Camag TLC Scanner-3, Camag Win CAT software with stationary phase precoated silica gel 60F ₂₅₄ and mobile phase consisting of ethyl acetate:toluene:methanol: ammonium hydroxide (1:6:2:0.1 v/v/v/v) were used. The detection of spot was carried out at 282 nm. The R f value was found to be 0.33±0.02. The methods were validated in terms of linearity, accuracy and precision. The linearity curves were found to be linear over 10-150 µg/ml for high performance thin layer chromatography and 100-600 ng/spot for high performance thin layer chromatography. The limit of detection and limit of quantification for high performance thin layer chromatography were found to be 2.0 and 10 µg/ml, respectively, and for the high performance thin layer chromatography, 30 and 100 ng/spot, respectively. The proposed methods were successfully used for estimation of nebivolol hydrochloride in tablet dosage form.