Clark C P, Snider B G, Bowman P B
Control Research and Development, Upjohn Company, Kalamazoo, MI 49001.
J Chromatogr. 1987 Nov 6;408:275-83. doi: 10.1016/s0021-9673(01)81810-0.
An isocratic high-performance liquid chromatographic (HPLC) method was developed to determine the enantiomeric purity of a benzindene prostaglandin (U-62,840) which is currently being evaluated for pharmaceutical applications. The enantiomers were converted to diastereomeric amide derivatives using optically pure S-(--)-1-phenylethylamine (greater than 99.9%). Separation of the diastereomers was demonstrated on achiral silica-based stationary phases using a hexane-dioxane-water mobile phase and UV detection at 214 nm. Due to the use of an optical derivatizing agent, method validation addressed the issues of optical purity of the reagent and comparative rates of reaction of each enantiomer. Analytical data is reported that demonstrates quantitative derivation, linearity, accuracy, precision, and ruggedness of the optimized assay conditions. Limit of quantitation for the enantiomeric impurity was determined to be 1.1% (signal-to-noise ratio 13). A brief description of the results of a parallel study of an enantiomeric separation using various chiral HPLC columns is included.
开发了一种等度高效液相色谱（HPLC）方法，用于测定一种苯茚二烯前列腺素（U-62,840）的对映体纯度，该前列腺素目前正在进行药物应用评估。使用光学纯的S-（-）-1-苯乙胺（大于99.9%）将对映体转化为非对映体酰胺衍生物。使用己烷-二氧六环-水流动相和214nm紫外检测，在手性硅胶固定相上实现了非对映体的分离。由于使用了光学衍生剂，方法验证涉及试剂的光学纯度以及每种对映体的相对反应速率问题。报告了分析数据，证明了优化分析条件下的定量衍生、线性、准确性、精密度和耐用性。对映体杂质的定量限测定为1.1%（信噪比13）。文中还简要介绍了使用各种手性HPLC柱进行对映体分离的平行研究结果。
