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Development and validation of UV and RP-HPLC methodfor estimation of Nepafenac in bulk drug and opthalmic formulation

Sadhana Rajput , Chhaya Shrimali , Madhuri Baghel


An ultraviolet (UV) and reverse phase high-performance liquid chromatographic (RP-HPLC) methods were developed and validated for the quantitative determination of Nepafenac in bulk and suspension formulation. UV method was performed with the λ max at 238.0 nm and 378.0 nm. RP-HPLC was carried out using C8 Olyster columnn with a mobile phase Acetonitrile: 10 mM Ammonium Formate buffer (pH 4.0 with formic acid): Methanol (27.5:45:27.5). The flow rate was 1.0 ml min-1 and effluent was detected at 238 nm. The retention time of NEPA was 4.60 min. Both the methods showed good linearity, reproducibility and precision. No spectral or chromatographic interferences from the excipients were found in UV and RP-HPLC. Validation parameters such as linearity, precision, accuracy, and specificity were determined. The methods were successfully applied to commercial formulation.

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