University of Belgrade, Faculty of Pharmacy, Department of Drug Analysis, Vojvode Stepe 450, Belgrade, Serbia.
University of Belgrade, Faculty of Pharmacy, Department of Drug Analysis, Vojvode Stepe 450, Belgrade, Serbia.
J Pharm Biomed Anal. 2023 Oct 25;235:115645. doi: 10.1016/j.jpba.2023.115645. Epub 2023 Aug 15.
In this paper, method for enantiomeric purity testing of fourth-generation fluoroquinolone, moxifloxacin hydrochloride, was developed and validated. Exceptional enantioselectivity for this assay was achieved using cyclodextrin type Chiral Stationary Phase (CSP), phenylcarbamate-β-cyclodextrin CSP, and mobile phase consisted of acetonitrile and triethylammonium acetate (TEAA) buffer. Analytical Quality by Design (AQbD) methodology, comprising Design of Experiments (DoE) - Design Space (DS) approach, was used for method development. In order to select appropriate Critical Method Parameters (CMPs), risk assessment was done using combined three step strategy that involved Ishikawa diagram - CNX (Control, Noise and eXperimental) - FMEA (Failure Mode and Effect Analysis). Three CMPs were further selected and investigated in this study: acetonitrile content in the mobile phase (30-50%, v/v), triethylamine content in the TEAA buffer (0.1-1.5%, v/v) and aqueous phase pH (3.5-4.5). Monte Carlo simulations were performed and 3D-DS was computed. One point situated in the center of 3D-DS was selected as working point for method validation, with the following values of CMPs: acetonitrile content in the mobile phase set to 37% (v/v), triethylamine content in TEAA 0.8% and pH value of the aqueous phase set at 4.0. Also, 2D-DS was created (with fixed one factor - pH value of aqueous phase at 4.0) which also gave us confirmation that the selection of working conditions was suitable. The proposed enantioselective method was further on tested for its quantitative robustness, as well as for its suitability for the intended purpose through validation studies.
本文开发并验证了第四代氟喹诺酮盐酸莫西沙星对映体纯度的检测方法。该分析采用环糊精型手性固定相（CSP），苯甲酰基-β-环糊精 CSP，以乙腈和三乙胺乙酸盐（TEAA）缓冲液为流动相，实现了对映体的出色选择性。分析质量源于设计（AQbD）方法学，包括实验设计（DoE）-设计空间（DS）方法，用于方法开发。为了选择合适的关键方法参数（CMPs），使用包含石川图-CNX（控制、噪声和实验）-失效模式与影响分析（FMEA）的三步综合风险评估策略进行了风险评估。在本研究中进一步选择和研究了三个 CMPs：流动相中乙腈的含量（30-50%，v/v），TEAA 缓冲液中三乙胺的含量（0.1-1.5%，v/v）和水相的 pH 值（3.5-4.5）。进行了蒙特卡罗模拟并计算了 3D-DS。在 3D-DS 的中心选择一个点作为方法验证的工作点，CMPs 的值如下：流动相中乙腈的含量设定为 37%（v/v），TEAA 中的三乙胺含量为 0.8%，水相的 pH 值设定为 4.0。此外，还创建了 2D-DS（固定一水相的 pH 值为 4.0），这也证实了工作条件的选择是合适的。通过验证研究，进一步测试了所提出的对映选择性方法的定量稳健性及其是否适合预期用途。
