R&D Analytical Chemistry Department, Pharmaceutical Research Institute, Rydygiera 8, 01-793 Warsaw, Poland.
R&D Analytical Chemistry Department, Pharmaceutical Research Institute, Rydygiera 8, 01-793 Warsaw, Poland.
J Pharm Biomed Anal. 2014 Mar;91:1-6. doi: 10.1016/j.jpba.2013.12.009. Epub 2013 Dec 19.
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.
本研究提出了一种应用高效液相色谱（HPLC）与分光光度紫外检测器分析奈帕芬酸（API）化学纯度和含量的方法。在文献检索中，仅发现了少量关于奈帕芬酸的出版物。HPLC-UV 方法主要出现在关于奈帕芬酸合成和化学纯度的专利文献中。所提出的方法能够分离奈帕芬酸的所有潜在相关化合物并定量奈帕芬酸的含量。由于药典中没有关于奈帕芬酸的专论，因此进行了完整的验证程序，使该方法能够在常规分析中使用。在开发阶段选择了流动相（10mM 甲酸铵，pH4.1）和 HPLC 柱（Phenomenex Gemini-NX C18）的组成。所呈现的数据证实了所开发方法的优势。四种最具潜力的杂质已被验证为定量测试，其余杂质已被验证为限量测试-根据 ICH Q2（R1）。化学纯度方法的准确度/回收率结果在 90-108%之间，在含量研究中为 99%-101%；检测限低至 15-30ng/mL。线性通过了所有统计检验。
