The Chromatography of Amino Acids on Ion Exchange Resins. Use of Volatile Acids for Elution
The Chromatography of Amino Acids on Ion Exchange Resins. Use of Volatile Acids for Elution
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离子交换树脂上氨基酸的色谱分析。
DOI：
10.1021/ja01652a059
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发表时间：
1954
影响因子：
15
通讯作者：
W. Stein
中科院分区：
文献类型：
--
作者：
C. Hirs;S. Moore;W. Stein
为了从蛋白质水解物中分离纯氨基酸，有可用的色谱系统是有利的，其中洗脱液可以通过简单的蒸发去除。先前以初步形式概述的方法，其中用HC1从磺化聚苯乙烯树脂（Dowex 50-X8）中洗脱氨基酸，已被证明可用于此目的，但分辨率不足以允许分离蛋白质水解物的所有常见成分。在本通讯中描述的程序中，天冬氨酸、谷氨酸和酪氨酸通过30厘米的水解液分离。Dowex 1-X8（一种强碱性树脂）的乙酸形式柱，使用0.5 N醋酸作为洗脱液，混合物的剩余组分在150 cm的隔板上分离。用1 ~ 4 Nhc1洗脱Dowex 50-X4柱。离子交换柱已在许多不同的方法中用于氨基酸的分离。为定量分析而开发的高分辨率色谱方法1·2可用于制备目的，如果通过在阳离子或阴离子树脂上循环流出物将氨基酸从非挥发性缓冲盐中分离出来。3.4例如，talan等人以这种方式从尿液中分离出氨基酸3-甲基组氨酸。6使用挥发性甲酸铵和醋酸盐缓冲液（可在40℃升华去除），最近设计了一种氨基酸分离系统，该系统也足够温和，原则上适用于制备肽或其他可能在酸溶液中不稳定的物质。如果不需要更高的洗脱分析分辨率，Partridge及其同事的位移开发方法往往具有优势，因为它们的容量更大。6
For the isolation of pure amino acids from protein hydrolysates, it is advantageous to have available chromatographic systems in which the eluants can be removed by simple evaporation. The method previously outlined in preliminary form, wherein amino acids are eluted from a sulfonated polystyrene resin (Dowex 50-X8) with HC1, has proved useful for this pur-pose, but the resolving power has not been sufficient to permit isolationof all of the common constituents of protein hydrolysates. In the procedure described in the present communication, aspartic acid, glutamic acid and tyrosine are isolated by passage of the hydrolysate over a 30-cm. column of the acetate form of Dowex 1-X8 (a strongly basic resin) using 0.5 N acetic acid as eluant, and the remaining components of the mixture are separated on a 150-cm. columnof Dowex 50-X4 by elution with 1 to 4 Nhc1.Ion-exchange columns have been utilized in a number of different ways for the isolation of amino acids. The chromatographic methods of high resolving power developed for quantitative analysis1· 2 can be used for preparative purposes if the amino acids are separated from thenon-volatile buffer salts by cycling the effluent over cationic or anionic res-ins. 3· 4 For example, the amino acid 3-methylhisti-dine was isolated from urine in this manner by Tallan, et al. 6 Using volatile ammonium formate and acetate buffers, which are removable by sublimation at 40, a system for the isolation of amino acids has been recently devised,® which is also sufficiently mild to be applicablein principle to the preparation of peptides or other substances that might be labile in acid solution. The displacement development methodsof Partridge and associates are fre-quently advantageous, in view of their greater capacity, if the higher resolving power of elution analysis isnot required. 6