Malik Poonam, Bhushan Ravi
Department of Chemistry, Indian Institute of Technology Roorkee, Roorkee, India.
Methods Mol Biol. 2019;2030:219-236. doi: 10.1007/978-1-4939-9639-1_17.
Enantioseparation studies of proteinogenic, non-proteinogenic, and dansyl amino acids are described herein by using liquid chromatographic techniques, i.e., HPLC and TLC. A researcher who wants to perform amino acid (AA) analysis or separate enantiomers of AAs by HPLC or TLC can follow the method. Figures included represent the actual experiments.Synthesis and application of chiral derivatizing reagents (CDRs) based on cyanuric chloride (CC) and difluorodinitrobenzene (DFDNB) have been described for AA analysis and enantioseparation by indirect approach. The methods represent pre-column derivatization of AAs and represent a good and less expensive substitute of AA analyzer. The application of commercial "Chiralplate" and use of erythromycin and L-tartaric acid have been described as chiral selector either as impregnating reagent in the stationary phase or as an additive in the mobile phase for direct enantioseparation by TLC. Application of the homemade TLC plates has also been described; the methods are successful in obtaining the native enantiomer as well.
本文描述了使用液相色谱技术,即高效液相色谱(HPLC)和薄层色谱(TLC)对蛋白质氨基酸、非蛋白质氨基酸和丹磺酰氨基酸进行对映体拆分研究。想要通过HPLC或TLC进行氨基酸(AA)分析或拆分AA对映体的研究人员可以遵循该方法。所附的图表展示了实际实验。基于三聚氯氰(CC)和二氟二硝基苯(DFDNB)的手性衍生试剂(CDR)的合成及应用已被描述用于通过间接方法进行AA分析和对映体拆分。这些方法代表了AA的柱前衍生化,是AA分析仪的一种良好且成本较低的替代方法。已描述了商业“手性板”的应用以及红霉素和L - 酒石酸作为手性选择剂在固定相中作为浸渍试剂或在流动相中作为添加剂用于通过TLC直接进行对映体拆分。还描述了自制TLC板的应用;这些方法也成功获得了天然对映体。