反相液相色谱中动力学因素对肽谱影响规律研究

Rule Effecting Peptide Mapping from Dynamic Factor in Reverse Phase Liquid Chromatography

研究了用反相色谱模式,在不同流动相流速条件下所得到的细胞色素C的肽谱图.发现在低流速时,保留弱的肽谱峰增多,而在高流速时保留强的肽谱峰增多,未发现溶质分子质量与溶质相对保留值(RRT)之间存在有特定的变化关系.然而却发现了两种变化规律,即溶质相对保留值(RRT)与流动相流速之间的双对数线性关系(1)log RRT=a+blogν,及所得该二线性参数,a与b之间的线性关系(2)a=c+db.预计该结果可能会在液相色谱指纹图谱研究和蛋白质组学中分离低丰度蛋白时有潜在的应用价值.

Different peptide mappings of cytochrome-C were obtained by reversed-phase liquid chromatography （RPLC） when the flow rate of mobile phase was changed. The result shows that the peak number of peptides with weak retention increases at a low flow rate of mobile phase and that the peak number of peptides with strong retention increases at a high flow rate of mobile phase. A specific relationship between the molecular mass of peptides and retention values was not found. However, two rules were found that （1） the logarithm of the relative retention time （RRT） of peptides was proportional to the logarithm of the flow rate of mobile phase （v）, i.e. log RRT=a＋blog v, and （2） the two linear parameters from the above equation obeyed the equation： a=c＋db. These results may have a potential usefulness for print finger map and proteomics.