Péter A, Tóth G, Olajos E, Tourwé D
Department of Inorganic and Analytical Chemistry, József Attila University, Szeged, Hungary.
J Chromatogr A. 1995 Jun 30;705(2):267-73. doi: 10.1016/0021-9673(95)00317-g.
A reversed-phase high-performance liquid chromatographic (RP-HPLC) method was developed to obtain pure erythro[2S3S, 2R3R]- and threo[2S3R, 2R3S]-beta-methylphenylalanine. These amino acids were incorporated into an enkephalin, H-Tyr-D-Ala-Gly-beta-MePhe-Val-Val-Gly-NH2, and into a deltorphin C, H-Tyr-D-Ala-beta-MePhe-Asp-Val-Val-Gly-NH2, analogue, which yielded four diastereoisomers of the peptides. The diastereoisomers were separated on different columns and with different eluent systems. The sequence of elution of the peptide diastereoisomers was determined after hydrolysis of the peptides. For identification of the beta-methylphenylalanine enantiomers, enzymatic degradation and an RP-HPLC method were used, with application of 1-fluoro-2,4-dinitrophenyl-5-L-alanine amide as derivatizing reagent.
开发了一种反相高效液相色谱（RP-HPLC）方法来获取纯的赤藓糖型[2S3S, 2R3R]-和苏阿糖型[2S3R, 2R3S]-β-甲基苯丙氨酸。这些氨基酸被掺入脑啡肽H-Tyr-D-Ala-Gly-β-MePhe-Val-Val-Gly-NH₂和强啡肽C类似物H-Tyr-D-Ala-β-MePhe-Asp-Val-Val-Gly-NH₂中，这产生了四种肽的非对映异构体。这些非对映异构体在不同的色谱柱上和使用不同的洗脱系统进行分离。在肽水解后确定肽非对映异构体的洗脱顺序。为了鉴定β-甲基苯丙氨酸对映体，使用了酶促降解和RP-HPLC方法，并应用1-氟-2,4-二硝基苯基-5-L-丙氨酸酰胺作为衍生试剂。
