Departamento de Química Analítica, Química Física e Ingeniería Química, Facultad de Ciencias, Universidad de Alcalá, Alcalá de Henares (Madrid), Spain.
Instituto de Investigación Química Andrés M. del Río (IQAR), Universidad de Alcalá, Alcalá de Henares (Madrid), Spain.
Electrophoresis. 2019 Aug;40(15):1951-1958. doi: 10.1002/elps.201900077. Epub 2019 May 31.
A MEKC methodology with UV detection was developed for the enantioselective separation of selenomethionine (SeMet). The use of (+)-1-(9-fluorenyl)ethyl chloroformate (FLEC) as chiral derivatization reagent to form SeMet diastereomers enabled their subsequent separation using ammonium perfluorooctanoate (APFO) as a volatile pseudostationary phase. The effect of APFO concentration and pH, temperature, injection volume, and derivatization conditions (time and FLEC/SeMet ratio) were evaluated in order to select the best separation conditions. A chiral resolution of 4.4 for DL-SeMet was achieved in less than 6 min using 100 mM APFO at pH 8.5 as electrophoretic buffer. Satisfactory results were obtained in terms of linearity, precision (RSD from 3.4 to 5.1% for migration times and from 1.8 to 4.6% for corrected peak areas), accuracy, and LODs (3.1 × 10 M and 3.7 × 10 M for d and l enantiomers, respectively). The method was successfully applied to the determination of l-SeMet in food supplements.
建立了一种 MEKC 方法，并结合紫外检测，用于手性拆分硒代蛋氨酸（SeMet）。采用（+）-1-（9-芴基）乙基氯甲酸酯（FLEC）作为手性衍生化试剂，形成 SeMet 非对映异构体，随后使用全氟辛烷铵（APFO）作为挥发性拟固定相进行分离。为了选择最佳分离条件，考察了 APFO 浓度和 pH 值、温度、进样体积以及衍生化条件（时间和 FLEC/SeMet 比）的影响。在 pH 8.5 的电泳缓冲液中使用 100 mM APFO，不到 6 分钟即可实现 DL-SeMet 的手性拆分，分离度为 4.4。该方法在迁移时间的线性度、精密度（RSD 为 3.4%至 5.1%）和校正峰面积的精密度（RSD 为 1.8%至 4.6%）、准确度和检出限（d 和 l 对映体分别为 3.1×10 和 3.7×10 ）方面均取得了满意的结果。该方法成功应用于食品补充剂中 l-SeMet 的测定。
