用反相液相色谱分离多肽时峰容量与色谱柱长度关系的研究

Studies on Relationship Between Peak Capacity and Column Length for Separation of Peptides by High Performance Liquid Chromatography

在蛋白质组学研究中,多肽混合物的有效分离对蛋白质鉴定和蛋白质之间相互作用的研究起着决定性的影响。基于此,用反相液相色谱研究了在两个不同长度的色谱柱上分离多肽混合物时色谱柱长度与峰容量的关系,同时考察了梯度洗脱时间对峰容量和峰宽的影响。实验结果表明,色谱柱长度对峰容量有显著的影响,而延长梯度洗脱时间不仅可以增加峰容量,而且可以增加峰宽。这说明用毛细管液相色谱 串联质谱联用方法对多肽混合物进行分离鉴定时,采用较长的色谱柱和较长的梯度洗脱时间有利于对更多的多肽进行分析鉴定。

In the study of proteomics, the high efficiency separation of peptides is very important for the identification of proteins and their interactions. Therefore, the relationship between peak capacity and column length for the separation of peptides on two reversed-phase columns with different lengths by reversed-phase high performance liquid chromatography was investigated and the effect of gradient times on peak capacity and peak width was examined. The experimental results showed that column length has significant effect on peak capacity.Furthermore, longer gradient times increase not only the peak number, but also the peak width, which is very favorable for the detection of more complicated mixture of peptides when they are separated and identified by capillary liquid chromatography combined with tandem mass spectrometry.