Reverse‐phase high‐performance liquid chromatography method has been developed for the determination of brivaracetam stereoisomeric impurities such as (R,S)‐brivaracetam, (R,R)‐brivaracetam, and (S,S)‐brivaracetam with good resolution using the chiral column, Chiral PAK IG‐U (100 × 3.0 mm; 1.6 μm). The method is simple, stability‐indicating, and compatible with LC–MS. The separation was achieved with the mobile phase consisted of 10 mM ammonium bicarbonate along with acetonitrile in an isocratic mode. The column temperature and wavelength were monitored at 40°C and 215 nm, respectively. The method showed adequate specificity, sensitivity, linearity, accuracy, precision, and robustness inline to ICH tripartite guidelines. The limit of detection and quantification limits were 0.3 and 0.8 μg ml⁻¹, respectively, for all stereoisomeric impurities and brivaracetam. The developed method was found to be linear over the concentration range of 0.8–5.6 μg ml⁻¹ for stereoisomeric impurities with a correlation coefficient >0.999. The method was precise (%RSD < 5.0), robust, and accurate (with 85%–115% recovery). The values of retention times of stereoisomeric impurities, (R,S)‐brivaracetam, (R,R)‐brivaracetam, and (S,S)‐brivaracetam, were 4.9, 5.4, and 6.6 min, respectively, and resolution among the impurities were 2.0, 3.3, and 4.7, respectively. In addition, forced degradation studies were performed to prove that method was stability‐indicating. The enrichment of isomeric impurity, (R,R)‐brivaracetam, was observed under basic stress conditions of brivaracetam and proposed a plausible mechanism to enhance that isomeric impurity. As well, a good separation among brivaracetam and its stereoisomeric impurity peaks was observed in the presence of degradation products and process‐related impurities.

中文翻译：

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开发一种有效的，经过验证的，有效的，可指示稳定性的HPLC方法，用于拆分溴乙酰胺的立体异构体杂质。

已经开发了反相高效液相色谱法，用于测定溴乙拉西坦的立体异构杂质，例如（R，S）-溴乙酰胺，（R，R）-溴乙酰胺和（S，S使用手性色谱柱Chiral PAK IG-U（100×3.0 mm; 1.6μm）具有良好的分离度。该方法简单，稳定，并与LC-MS兼容。分离是通过以等度模式由10 mM碳酸氢铵和乙腈组成的流动相完成的。分别在40°C和215 nm处监测色谱柱温度和波长。该方法显示出足够的特异性，灵敏性，线性，准确性，精密度和鲁棒性，符合ICH三方准则。所有立体异构杂质和溴西西坦的检出限和定量限分别为0.3和0.8μgml ^-1。发现所开发的方法在0.8–5.6μgml ^-1的浓度范围内是线性的相关系数> 0.999的立体异构体杂质。该方法准确（％RSD <5.0），稳健且准确（回收​​率85％–115％）。立体异构体杂质（R，S）-溴乙酰胺，（R，R）-溴乙酰胺和（S，S）-溴乙酰胺的保留时间分别为4.9，5.4和6.6分钟，杂质之间的分离度分别为2.0、3.3和4.7。此外，进行了强制降解研究以证明该方法具有稳定性。异构体杂质的富集（R，R）-brivaracetam，在布立西坦的基本应力条件下观察到，并提出了增加该异构体杂质的合理机制。同样，在存在降解产物和过程相关杂质的情况下，观察到溴氰西坦及其立体异构体杂质峰之间的良好分离。