Abstract
A high-performance thin layer chromatography and first order derivative UV-visible spectroscopic method was developed and validated for estimation of pimobendan (PIMO) in pharmaceutical tablet dosage form. A simple, selective, precise, and stability-indicating high performance thin-layer chromatographic (HPTLC) method for the analysis of PIMO was developed and validated. The method employed HPTLC aluminium plates precoated with silica gel 60F-254 as the stationary phase. The solvent systems consist of Ethyl Acetate: Toluene: Methanol (5:4:1 v/v). The system was found to give a compact band for PIMO (Rf value of 0.49 ± 0.02). Densitometric analysis of PIMO was carried out in the absorbance mode at 254 nm. The linear regression analysis data for the calibration plots showed good linear relationship with r2 = 0.999 with respect to peak area in the concentration range 0.5–3.0 ng/spot. The method was validated as per ICH guidelines. The LOD and LOQ for PIMO were 0.059 and 0.18ng / band. In spectroscopic method the λ max of PIMO in methanol was found to be 328 nm. The same spectrum was derivatised in to first order derivative; wherein it showed maximum amplitude of the trough at 349 nm. This wavelength was selected for analysis. The drug follows linearity in the concentration range 10-60 μg/ml with correlation coefficient value 0.999. Method was validated according to the ICH guidelines. The proposed method was successfully used for quantitative analysis of tablets. No interference from any component of pharmaceutical dosage form was observed. Validation studies revealed that method is specific, rapid, reliable, and reproducible. The high recovery and low relative standard deviation confirm the suitability of the method for routine determination of PIMO in bulk drug and tablet dosage form.
