色氨酸在 Toyopearl HW-40S 凝胶填料上非理想排阻行为的研究

发现１８种氨基酸特别是色氨酸．在ＴｏｙｏｐｅａｒｌＨＷ－４０Ｓ树脂所填色谱柱上表现出非理想排阻行为的基础上，以色氨酸在该柱上的保留时间为指标，探讨不同流动相改良剂对该凝胶与色氨酸之间的相互作用，阐明这种反常行为是疏水作用，离子交换以及氢键三者综合作用的结果，而疏水作用是最主要的影响因素．实验结果表明，流动相组成对该凝胶色谱行为具有明显的影响，从而有助于确立该凝胶理想的排阻色谱条件，为利用该凝胶所具有的吸附性能分离分析肽类和氨基酸等小分子物质奠定了基础

Deviation of Size-Exclusion Chromatographic Behavior of Tryptophan on Toyopearl HW-40S Column

A deviation of size-exclusion chromatographic behavior of animo acids on Toyopearl HW-40S column was reported, on the column 18 animo acids were not eluted in the same fraction in the total volume, but separated into 5 fractions with tryptophan as the last on far behind the total volume. Interaction between tryptophan and the matrices was investigated under different conditions in order to elucidate the chemical nature of the deviation. It was found that addition of trifluoroacetic acid and sodium chloride of high concentration in mobile phase, dramatically increased the retention time of trypophan, while addition of acetonitrile, sodium chloride of lower concentration and hydrogen bond breaker urea, reduced the interaction. It is concluded that adsorption of tryptophan to Toyopearl HW-40S is basically due to the hydrophobic nature with limited influence of hydrogen bond and ionic interaction. The former factor can be eliminated by keeping acentonitrile concentration at above 50%(V/V), while the latter two can be reduce to the minimum by adding 0.17 mol/L sodium chloride and 3.51 mol/L urea. The result not only lead to better understanding of size-exclusion chromatography of this matrices, but also help to apply hydrophobic chromatography of this matrices to the isolation of animo acids and peptides.