title: Development and validation of high performance liquid chromatography with a spectrophotometric detection method for the chemical purity and assay of nepafenac.
abstract: The study is a proposition of the application of high performance liquid chromatography (HPLC) with a spectrophotometric UV range detector to analyze the chemical purity and assay of nepafenac, an active pharmaceutical ingredient (API). During literature search only a few publications were found about nepafenac. HPLC UV methods were mainly presented in patent documents about nepafenac synthesis and chemical purity. The presented method allows to separate all potential related compounds from nepafenac and to quantitate the nepafenac amount. As there is no official monograph in the pharmacopeias about nepafenac, the performed full validation procedure makes the method ready to use in routine analysis. The composition of the mobile phase (10mM ammonium formate, pH 4.1) and the HPLC column (Phenomenex Gemini-NX C18) were selected during the development step. Presented data confirm the benefits of the developed method. Four of the most potential impurities were validated as for the quantitative test and the rest of impurities were validated as for the limit test - according to ICH Q2(R1). The accuracy/recovery results for the chemical purity method are within 90-108%, in the case of assay studies from 99% to 101%; the limit of detection is as low as 15-30ng/mL. The linearity passes all statistical tests.
authors.affiliation: R&D Analytical Chemistry Department, Pharmaceutical Research Institute, Rydygiera 8, 01-793 Warsaw, Poland. Electronic address: e.lipiec@ifarm.eu.
authors.forename: Elżbieta
authors.initials: E
authors.lastname: Lipiec-Abramska
authors.affiliation: R&D Analytical Chemistry Department, Pharmaceutical Research Institute, Rydygiera 8, 01-793 Warsaw, Poland.
authors.forename: Łukasz
authors.initials: Ł
authors.lastname: Jedynak
authors.affiliation: Chemistry Department, Pharmaceutical Research Institute, Rydygiera 8, 01-793 Warsaw, Poland.
authors.forename: Adam
authors.initials: A
authors.lastname: Formela
authors.affiliation: Minisynthesis Department, Pharmaceutical Research Institute, Rydygiera 8, 01-793 Warsaw, Poland.
authors.forename: Jacek
authors.initials: J
authors.lastname: Roszczyński
authors.affiliation: Chemistry Department, Pharmaceutical Research Institute, Rydygiera 8, 01-793 Warsaw, Poland.
authors.forename: Marcin
authors.initials: M
authors.lastname: Cybulski
authors.affiliation: R&D Analytical Chemistry Department, Pharmaceutical Research Institute, Rydygiera 8, 01-793 Warsaw, Poland.
authors.forename: Maria
authors.initials: M
authors.lastname: Puchalska
authors.affiliation: R&D Analytical Chemistry Department, Pharmaceutical Research Institute, Rydygiera 8, 01-793 Warsaw, Poland.
authors.forename: Joanna
authors.initials: J
authors.lastname: Zagrodzka
mesh_terms: D044482:Benzeneacetamides / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004340:Drug Contamination; D010648:Phenylacetates / Q000737:chemistry*; D013053:Spectrophotometry / Q000379:methods*
chemical_list: D044482:Benzeneacetamides; D010648:Phenylacetates; C414203:nepafenac
keywords: HPLC UV method; MS compatible; Method validation; Nepafenac assay; Nepafenac impurities