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以C8 柱为固定相 ,研究了微管结合蛋白2中两段重要肽段(肽1 ,肽2)和猪胰岛素在反相色谱上的保留行为。当以甲醇溶液为流动相时 ,猪胰岛素保留因子的对数值(lnK′)随流动相有机溶剂体积分数(φ)的变化呈现很好的线性关系。当以乙腈为流动相测定肽1的lnK′随 φ变化关系时 ,所得结果显示lnK′与 φ呈现良好的二次曲线关系。以乙腈为流动相测定肽1和肽2的Van’tHoff曲线(lnK′与1/T的关系图)时 ,发现两种多肽的Van’tHoff曲线分别呈二次和三次方曲线。所有这些结果说明多肽和蛋白质在反相色谱中保留行为的复杂性。
Using C8 column as the stationary phase, the retention behavior of two important peptides (peptide 1, peptide 2) and porcine insulin in microtubule-binding protein 2 was studied by reverse phase chromatography. When methanol solution was used as mobile phase, the logarithmic value (lnK ’) of porcine insulin retention factor showed a good linear relationship with the volume fraction of mobile phase organic solvent (φ). When the lnK ’of peptide 1 changes with the change of φ with acetonitrile as the mobile phase, the obtained results show that lnK’ and φ exhibit a good quadratic curve. When the Van’t Hoff curve (lnK ’versus 1 / T) of peptide 1 and peptide 2 was measured using acetonitrile as the mobile phase, the Van’t Hoff curves of the two polypeptides were found to be quadratic and cubic curves, respectively. All of these results illustrate the complexity of retaining the behavior of peptides and proteins in reversed-phase chromatography.