Wang Fangjun, Ye Mingliang, Dong Jing, Tian Ruijun, Hu Lianghai, Han Guanghui, Jiang Xinning, Wu Ren'an, Zou Hanfa
National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China.
J Sep Sci. 2008 Aug;31(14):2589-97. doi: 10.1002/jssc.200800181.
The postcolumn void volume, which is introduced by the connecting tubing and void ESI emitter in the nanoflow LC coupled with MS/MS system (microLC-MS/MS), is harmful for the analysis of peptides in shotgun proteome analysis. A new type of porous C12 monolithic ESI emitter was prepared to eliminate the disruption and mixing effects occurring in the connecting tubing and void emitter. It was demonstrated that the porous hydrophobic monolith inside the emitter played a key role in retaining the good peak profile, and the average peak capacity of the whole separation system increased 12.8% in contrast to commercially available void emitter. Then, the porous C12 monolithic emitter was applied in label-free quantitative proteome analysis of two standard protein mixtures that were spiked into the tryptic digest of mouse livers extract. Compared to commercially available void ESI emitter, the number of proteins with reliable results in quantification increased greatly. And the relative quantities of the four standard proteins were all determined with the relative error < or = 6.8%. However, quantitative information of only three standard proteins could be obtained when void emitter was used.
在纳流液相色谱与串联质谱系统（微液相色谱-串联质谱，microLC-MS/MS）联用中，连接管和空的电喷雾离子化（ESI）发射器所引入的柱后死体积，对鸟枪法蛋白质组分析中的肽段分析有害。制备了一种新型的多孔C12整体式ESI发射器，以消除连接管和空发射器中出现的干扰和混合效应。结果表明，发射器内部的多孔疏水整体在保持良好峰形方面起关键作用，与市售的空发射器相比，整个分离系统的平均峰容量提高了12.8%。然后，将多孔C12整体式发射器应用于两种标准蛋白质混合物的无标记定量蛋白质组分析，这两种混合物被添加到小鼠肝脏提取物的胰蛋白酶消化物中。与市售的空ESI发射器相比，定量结果可靠的蛋白质数量大幅增加。并且四种标准蛋白质的相对含量测定的相对误差均≤6.8%。然而，使用空发射器时只能获得三种标准蛋白质的定量信息。
