Cheng H, Shao T
Dalian Institute of Chemical Physics, the Chinese Academy of Sciences, Dalian, 116023.
Se Pu. 1997 Sep;15(5):405-7.
The hydrolytic solution of proteins mainly contains free amino acids and peptides. The separation of amino acids and di- and tri-peptides is very significant and also a complicated work. This report presents a chelated metal ion affinity chromatographic (CMAC) method for the separation of alpha-amino acids and peptides. Sephadex G10 was used as the solid matrix. It was epoxy-activated by epichlorophydrin; then coupled with iminodiacetate (IDA) and chelated with copper ion to produce immobilized copper-ion affinity chromatographic packing. Some examples are given for the chromatography of model mixtures of L-Val, L-His, L-Tyr, L-Try, Tyr-Try dipeptides and protein hydrolyzing solution of fish. The separation was based on the different stabilities of copper complexes of alpha-amino acids, peptides and IDA-Sephadex G10. The components which form weak complexes with copper apparently move along with the solvent front. Alpha-amino acids-copper complexes with a stability comparable to that of copper-IDA-Sephadex G10 are retained on the matrix. Peptides form strong complexes and catch copper from the matrix. They are only slightly retained. The results showed that alpha-amino acids and peptides were completely separated under the experimental conditions.
蛋白质水解液主要包含游离氨基酸和肽。氨基酸以及二肽和三肽的分离非常重要，同时也是一项复杂的工作。本报告介绍了一种用于分离α-氨基酸和肽的螯合金属离子亲和色谱法（CMAC）。使用葡聚糖凝胶G10作为固体基质。它通过环氧氯丙烷进行环氧活化；然后与亚氨基二乙酸（IDA）偶联并与铜离子螯合，以制备固定化铜离子亲和色谱填料。给出了一些关于L-缬氨酸、L-组氨酸、L-酪氨酸、L-色氨酸、酪氨酸-色氨酸二肽模型混合物以及鱼蛋白水解液色谱分析的示例。分离基于α-氨基酸、肽与IDA-葡聚糖凝胶G10的铜配合物稳定性的差异。与铜形成弱配合物的组分显然会随着溶剂前沿移动。与铜形成稳定性与铜-IDA-葡聚糖凝胶G10相当的配合物的α-氨基酸会保留在基质上。肽形成强配合物并从基质中捕获铜。它们仅被轻微保留。结果表明，在实验条件下α-氨基酸和肽被完全分离。
