PDF(Pubmed)
Abstract：
A reversed-phase high-performance liquid chromatographic (HPLC) method was developed for the simultaneous determination of the potential impurities of dexketoprofen, including the distomer R-ketoprofen. After screening the separation capability of four polysaccharide columns (Lux Amylose-1, Lux Amylose-2, Lux Cellulose-1 and Lux Cellulose-2) in polar organic and in reversed-phase modes, appropriate enantioseparation was observed only on the Lux Amylose-2 column in an acidified acetonitrile/water mixture. A detailed investigation of the mobile phase composition and temperature for enantio- and chemoselectivity showed many unexpected observations. It was observed that both the resolution and the enantiomer elution order can be fine-tuned by varying the temperature and mobile phase composition. Moreover, hysteresis of the retention times and enantioselectivity was also observed in reversed-phase mode using methanol/water mixtures on amylose-type columns. This could indicate that the three-dimensional structure of the amylose column can change by transitioning from a polar organic to a reversed-phase mode, which affects the enantioseparation process. Temperature-dependent enantiomer elution order and rare enthalpic/entropic controlled enantioseparation in the operative temperature range were also observed in reversed-phase mode. To find the best methodological conditions for the determination of dexketoprofen impurities, a full factorial optimization design was performed. Using the optimized parameters (Lux Amylose-2 column with water/acetonitrile/acetic acid 50/50/0.1 (v/v/v) at a 1 mL/min flow rate at 20 °C), baseline separations were achieved between all compounds within 15 min. Our newly developed HPLC method was validated according to the current guidelines, and its application was tested on commercially available pharmaceutical formulations. According to the authors\' knowledge, this is the first study to report hysteretic behavior on polysaccharide columns in reversed-phase mode.
摘要：
开发了一种反相高效液相色谱（HPLC）方法，用于同时测定右酮洛芬的潜在杂质，包括口服R-酮洛芬.在极性有机和反相模式下筛选了四个多糖柱（Lux直链淀粉-1，Lux直链淀粉-2，Lux纤维素-1和Lux纤维素-2）的分离能力之后，仅在LuxAmylose-2柱上在酸化的乙腈/水混合物中观察到适当的对映体分离。对映和化学选择性的流动相组成和温度的详细研究显示了许多意想不到的观察结果。观察到，可以通过改变温度和流动相组成来微调拆分和对映体洗脱顺序。此外，在直链淀粉型柱上使用甲醇/水混合物在反相模式下也观察到保留时间和对映选择性的滞后。这可能表明直链淀粉柱的三维结构可以通过从极性有机模式转变为反相模式而改变，影响对映分离过程。在反相模式下还观察到温度依赖性对映异构体洗脱顺序和在工作温度范围内罕见的焓/熵控制的对映异构体分离。寻找测定右酮洛芬杂质的最佳方法学条件,进行了全因子优化设计。使用优化的参数（LuxAmylose-2色谱柱，水/乙腈/乙酸50/50/0.1（v/v/v），流速为1mL/min，20°C），所有化合物之间在15分钟内实现基线分离。我们新开发的HPLC方法根据现行指南进行了验证，并在市售药物制剂上测试了其应用。根据作者的知识，这是首次报道反相模式下多糖柱的滞后行为的研究。
