title: Development and validation of HPLC analytical method for nepafenac in ophthalmic dosage form (suspension).
abstract: The aim of the present study was to develop and validate an analytical method for the estimation of nepafenac as a raw material as well as in dosage form (suspension) by using reverse phase high performance liquid chromatographic (RP-HPLC). The target was to obtain an easy, rapid, reproducible as well as a rugged method. The HPLC system that was used in the proposed study was LC-20AD liquid chromatograph equipped with SPD-20A UV-VIS detector. The separation was performed on C18 column which was attached with loop 20 β l. Elution was done at ambient temperature with a mobile phase consisting of acetonitrile: Water (40: 60v/v) at a flow rate of 1ml/min and at a wavelength of 254 nm. The proposed method was validated as per the ICH guidelines. The retention time for nepafenac was 7.49 minutes (% CV=0.0076). The percentage coefficient variation (CV) of six consecutive peak areas of injections was 0.34% with tailing factor 1.76. The peak area responses were linear within the concentration range of 0.078-20.0 βg/ml (R(2)=0.9993). The sensitivity of the method could be evaluated by limits of detection (LOD) (0.0195 β g/ml) and limits of quantitation (LOQ) (0.039 β g/ml). Nepafenac drug is s in its diluent that could see by intra-day (% CV =0.45-1.96) and inter-day variation (%CV=0.173-1.898%). The accuracy and recovery results of 80%, 100% and 120% were 97.40% to 102.10% with % CV of 0.3201% to 1.3496%. The robustness and ruggedness of the method are significantly broader and is reproducible. It could be used as a more convenient, efficient, easy and time saving method for the analysis of drug in raw material as well as in dosage form (ophthalmic suspension).
authors.affiliation: RAK College of Pharmaceutical Sciences, RAK Medical and Health Sciences University, Ras Al Khaimah, UAE / Department of Pharmaceutics, Faculty of Pharmacy, University of Karachi, Karachi, Pakistan.
authors.forename: Shahnaz
authors.initials: S
authors.lastname: Usman
authors.affiliation: Department of Pharmaceutics, Faculty of Pharmacy, University of Karachi, Karachi, Pakistan.
authors.forename: Muhammad
authors.initials: M
authors.lastname: Akram
authors.affiliation: Department of Chemistry, Faculty of Science, University of Karachi, Karachi, Pakistan.
authors.forename: Asif
authors.initials: A
authors.lastname: Aziz
authors.affiliation: RAK College of Pharmaceutical Sciences, RAK Medical and Health Sciences University, Ras Al Khaimah, UAE.
authors.forename: Venkat
authors.initials: V
authors.lastname: Ramesh
authors.affiliation: RAK College of Pharmaceutical Sciences, RAK Medical and Health Sciences University, Ras Al Khaimah, UAE.
authors.forename: Omar Abdulraheem
authors.initials: OA
authors.lastname: Sarheed
mesh_terms: D060433:Administration, Ophthalmic; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000008:administration & dosage / Q000032:analysis*; D044482:Benzeneacetamides / Q000008:administration & dosage / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D056148:Chromatography, Reverse-Phase; D004355:Drug Stability; D057230:Limit of Detection; D016014:Linear Models; D009883:Ophthalmic Solutions; D010648:Phenylacetates / Q000008:administration & dosage / Q000032:analysis*; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet; D013678:Technology, Pharmaceutical / Q000379:methods*
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D044482:Benzeneacetamides; D009883:Ophthalmic Solutions; D010648:Phenylacetates; C414203:nepafenac