On-line parallel reversed phase two-dimensional liquid chromatography for high-throughput analysis of complex proteomic samples  

作　　者：WANG Zhicong1,3, ZHANG Qinghe2, LI Tong2, ZHAO Zhongyi3 & ZHANG Weibing1 1. Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116011, China 2. Dalian Elite Analytical Instrument Co., Ltd., Dalian 116011, China 3. Faculty of Material Science and Chemical Engineering, China University of Geosciences, Wuhan 430074, China 

出　　处：《Science China Chemistry》2006年第6期527-533,共7页中国科学（化学英文版）

基　　金：Financial support from the National Natural Science Foundation of China (Grant Nos. 20175029 and 20375040) is gratefully acknowledged.

摘　　要：A comprehensive two-dimensional liquid chromatographic system (2D SCX/RP) is con- structed with a 10-port-2-way valve using strong cation exchange chromatography (Hypersil SCX, 100 mm×4.6 mm I.D.) followed by reversed phase chromatography (Hypersil BDS C18, 15 mm×4.6 mm I.D.) to separate the complex peptides from globin peptic hydrolysate. After the sample was loaded on the SCX column, the phosphate buffer (pH 4.0) was used to elute the peptides. Then, elutes flowed through the interface and the peptides focused on the head of the trapping columns (Hypersil BDS C18, 15 mm×4.6 mm I.D.) but salt passed into the waste. After the valve was switched, the samples were flushed with a backward flow into the RP analytical column. The peptides on the SCX were eluted with 12 discontinuous steps linearly increasing salt concentrations. The peptides enriched on the trapping column were desalted and separated by the RP columns. The resolution and the resolved peaks of the 2D SCX/RP system were greatly increased and the total peak capacity reached as high as 2280.A comprehensive two-dimensional liquid chromatographic system (2D SCX/RP) is con- structed with a 10-port-2-way valve using strong cation exchange chromatography (Hypersil SCX, 100 mm×4.6 mm I.D.) followed by reversed phase chromatography (Hypersil BDS C18, 15 mm×4.6 mm I.D.) to separate the complex peptides from globin peptic hydrolysate. After the sample was loaded on the SCX column, the phosphate buffer (pH 4.0) was used to elute the peptides. Then, elutes flowed through the interface and the peptides focused on the head of the trapping columns (Hypersil BDS C18, 15 mm×4.6 mm I.D.) but salt passed into the waste. After the valve was switched, the samples were flushed with a backward flow into the RP analytical column. The peptides on the SCX were eluted with 12 discontinuous steps linearly increasing salt concentrations. The peptides enriched on the trapping column were desalted and separated by the RP columns. The resolution and the resolved peaks of the 2D SCX/RP system were greatly increased and the total peak capacity reached as high as 2280.

关 键 词：two-dimensional liquid chromatography  interface  TRAPPING column  proteomics  globin. 

分 类 号：O657.72[理学—分析化学]