强阳离子交换色谱分离多肽混合物的条件优化  

Optimization of conditions for the separation of peptide mixtures using strong cation exchange chromatography

作　　者：王璐[1] 周兰兰[1] 钱小红[2] 张养军[2] 

机构地区：[1]安徽医科大学,安徽合肥230032 [2]蛋白质组学国家重点实验室军事医学科学院放射与辐射医学研究所北京蛋白质组研究中心,北京102206

出　　处：《色谱》2010年第4期368-373,共6页Chinese Journal of Chromatography

基　　金：国家重点基础研究规划项目(2006CB910803;2007CB914104);国家自然科学基金项目(30621063;20635010;20735005;20875101);蛋白质组学国家重点实验室项目(SKLP-K200807;SKLP-O200808)

摘　　要：以酵母全蛋白提取液的胰蛋白酶切产物为研究对象,对多维色谱分离中常用的强阳离子交换色谱的分离条件,包括上样量、盐的种类、调节缓冲液pH值的酸的种类及有机溶剂的比例进行了考察和比较。结果表明,在强阳离子交换色谱分离线性上样量范围内,在氯化铵溶液作为流动相,用磷酸调节流动相缓冲液pH值至2.7,且流动相中乙腈的体积分数为30%条件下进行梯度洗脱时,可获得最佳的分离结果。本结果可为采用二维色谱-质谱策略分析多肽混合物时的第一维强阳离子交换色谱分离条件的选择提供有益的参考。The separation conditions for strong cation exchange chromatography were investigated and compared using tryptic digest of protein extract from yeast.The conditions included the loading amount of the sample,the type of the salt in mobile phase,the type of the acid used to adjust the pH of the mobile phase and the proportion of organic solvent added in the elution solution.The experimental results indicated that high separation efficiency can be obtained by using ammonium chloride solution as optimal mobile phase,phosphoric acid to adjust the pH of mobile phases and 30%（v/v） organic solvent in mobile phases.The results suggested that a beneficial reference was presented for the selection of separation conditions of strong cation exchange chromatography when applying in two-dimensional chromatography-mass spectrometry strategy in proteomic research.

关 键 词：强阳离子交换色谱 蛋白质组学 多肽分离 

分 类 号：O658[理学—分析化学]