For the purpose of identifying enantiomeric impurities in the drug substance and pharmaceutical dosage forms of the novel anti-TB medication pretomanid, an RP-high-performance liquid chromatography method was devised. To ensure the robustness of the optimized approach, analytical quality by design was used. Studies on factor screening and risk evaluation helped pinpoint the critical method parameters (CMPs); resolution (R1), analyte retention time (R2) and tailing factor (R3) are those terms. Pareto charts, half-normal plots, 3D surface plots, 2D contour plots and 3D cube plots were used to study the effects of solo and interactive CMPs on critical analytical attributes. Analysis of variance (ANOVA) was used to verify the technique parameters’ confirmation of significance (P = 0.05). With a Chiral Cel OJ-3R (150 × 4.6 mm, 3 µm) and a mobile phase consisting of 20 mM of ammonium trifluoroacetate, pH = 2.5, and acetonitrile in a gradient mode, chromatographic separation was accomplished. At 30°C and 330 nm, the column’s temperature and wavelength, respectively, were recorded. The procedure is stability-indicating and is LC–MS compatible. According to the International Conference on Harmonization tripartite guidelines, the method demonstrated appropriate specificity, sensitivity, linearity, accuracy, precision and robustness. The LOD and LOQ were, respectively, 0.09 and 0.3 μg/mL. With a correlation coefficient of >0.990, it was discovered that the established method for enantiomeric impurity was linear over the concentration range of 0.3–2.25μg/mL. The approach exhibits adequate accuracy (%recovery = 85–115%), robustness (%RSD = 5.0) and precision (%RSD = 5.0). The method was also shown to be stability-indicating and was shown to provide effective separation in the presence of degradation products through the use of forced degradation tests.

中文翻译：

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Pretomanid 原料药和药物剂型中对映体杂质含量测定方法的开发、稳健性和验证方法的设计专家

为鉴定新型抗结核药物pretomanid原料药和药物剂型中的对映体杂质，设计了反相高效液相色谱法。为了确保优化方法的稳健性，使用了分析质量设计。因素筛选和风险评估研究有助于确定关键方法参数 (CMP)；分辨率 (R1)、分析物保留时间 (R2) 和拖尾因子 (R3) 就是这些术语。Pareto 图、半正态图、3D 曲面图、2D 等高线图和 3D 立方图用于研究单独和交互式 CMP 对关键分析属性的影响。方差分析 (ANOVA) 用于验证技术参数的显着性确认 (P = 0.05)。使用手性 Cel OJ-3R（150 × 4.6 mm，3 µm)和由20 mM三氟乙酸铵组成的流动相，pH = 2.5，和乙腈在梯度模式下，完成色谱分离。在 30°C 和 330 nm 下，分别记录色谱柱的温度和波长。该程序具有稳定性指示性，并且与 LC-MS 兼容。根据国际协调会议三方指南，该方法显示出适当的特异性、灵敏度、线性、准确度、精密度和稳健性。LOD 和 LOQ 分别为 0.09 和 0.3 μg/mL。相关系数 > 0.990，发现建立的对映体杂质的方法在 0.3–2.25μg/mL 的浓度范围内呈线性。该方法具有足够的准确度 (%recovery = 85–115%)、稳健性 (%RSD = 5.0) 和精密度 (%RSD = 5. 0). 该方法还显示出稳定性指示性，并显示通过使用强制降解测试可在存在降解产物的情况下提供有效分离。