对近年来蛋白质组研究中的多维液相色谱技术进行了系统综述,详细介绍了在线及离线式阀切换接口的DALPC(direct analysis of large protein complexes)技术及整体式无接口的MudPIT(multidimensional protein identifica-tion technology)系统,也系统阐述了其在蛋白质组研究中的应用。引用文献50篇。
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.
WANG Zhicong1,3, ZHANG Qinghe2, LI Tong2, ZHAO Zhongyi3 & ZHANG Weibing1 1. Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116011, China
A novel on-line parallel-trap supercritical fluid extraction(SFE)-high performance liquid chromatography(HPLC)system was established.The interfere consisted of automated ten-port valve and parallel traps were used to carry out continuous and alternate trapping and transferring of the sample extracted with supercritical carbon dioxide,and HPLC was used to analyze the transferred sample.The novel on-line system was used to monitor the extraction process of fruiting bodies of Ganoderma lucidum.The extracts of six consecutive cycles were analyzed and the optimum extraction time was determined,which accorded with the result obtained with off-line extraction.Using this system,the whole process can be monitored and the optimum extraction time can be determined with single run.In addition,it has many other advantages,such as low sample loading,high sensitivity and accuracy,short analysis time,avoiding sample pollution and easily operation etc.The system shows a great potential in the extraction and analysis of the complicated samples including traditional Chinese medicine and environmental samples etc.
