Finšgar Matjaž, Perva-Uzunalić Amra, Behr Heiko, Ledinek Nina, Knez Željko, Novak Zoran
Faculty of Chemistry and Chemical Engineering, University of Maribor, Smetanova ulica 17, 2000 Maribor, Slovenia.
Phenomenex, Ltd., Zeppelinstrasse 5, 63741 Aschaffenburg, Germany.
ACS Omega. 2020 Mar 30;5(14):7987-8000. doi: 10.1021/acsomega.0c00037. eCollection 2020 Apr 14.
Prednisolone, an important active pharmaceutical ingredient, is a synthetic glucocorticoid used for the preparation of various pharmaceutical products with anti-inflammatory and immunosuppressive properties. It is a challenge in high-performance liquid chromatography (HPLC) to separate the prednisolone peak and its structurally related substance (hydrocortisone), which only differs in a double bond at the C-1 position. Successful application of the HPLC method according to the European Pharmacopoeia monograph for related substances of prednisolone is very often limited to the chromatographic system available. This is due to the nonbaseline separation of the prednisolone and hydrocortisone peaks, which is strongly influenced by the instrument parameters and the chosen C18 column. First, an adjusted European Pharmacopoeia method for related substances of prednisolone was developed within the allowable adjustments. Next, an improved stability-indicating reversed-phase HPLC method for related substances of prednisolone was developed and validated for use in quality control laboratories for routine analysis. The optimized separation was performed on a Phenomenex Gemini C18 column (150 mm × 4.6 mm, 3 μm) using a gradient mobile-phase system consisting of acetonitrile/tetrahydrofuran/water (15:10:75 v/v/v), acetonitrile/water (80:20 v/v), and ultraviolet detection at 254 nm. A baseline separation was achieved, and stability indicating capability was demonstrated by a forced degradation study. A full validation procedure was performed in accordance with International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use guidelines.
泼尼松龙是一种重要的活性药物成分，是一种合成糖皮质激素，用于制备具有抗炎和免疫抑制特性的各种药品。在高效液相色谱法（HPLC）中，分离泼尼松龙峰及其结构相关物质（氢化可的松）是一项挑战，它们仅在C-1位的一个双键上有所不同。按照欧洲药典中泼尼松龙相关物质专论的HPLC方法，其成功应用常常局限于现有的色谱系统。这是因为泼尼松龙和氢化可的松峰的分离不是基线分离，这受到仪器参数和所选C18柱的强烈影响。首先，在允许的调整范围内开发了一种调整后的欧洲药典泼尼松龙相关物质方法。接下来，开发并验证了一种改进的用于泼尼松龙相关物质的稳定性指示反相HPLC方法，供质量控制实验室用于常规分析。优化的分离在Phenomenex Gemini C18柱（150 mm×4.6 mm，3μm）上进行，使用由乙腈/四氢呋喃/水（15:10:75 v/v/v）、乙腈/水（80:20 v/v）组成的梯度流动相系统，并在254 nm处进行紫外检测。实现了基线分离，并通过强制降解研究证明了其稳定性指示能力。按照人用药品注册技术要求国际协调会议指南进行了全面的验证程序。
