Tůma Petr
Institute of Biochemistry, Cell and Molecular Biology, Third Faculty of Medicine, Charles University, Prague 10, Czech Republic.
J Sep Sci. 2014 Apr;37(8):1026-32. doi: 10.1002/jssc.201400044. Epub 2014 Mar 13.
A laboratory-made INSTCoated fused-silica capillary has been newly used for CE separation of four mixtures of proteins in sodium phosphate BGEs at pH 3.0 and 2.5, respectively. The obtained separation efficiencies range from 145,000 theoretical plates per meter for myoglobin to 1,216,000 m(-1) for lysozyme. A total of 49-89% of the number of theoretical plates was obtained in a commercial polyvinyl alcohol coated capillary compared to the INSTCoated capillary under the same experimental conditions, 0-86% was obtained in a laboratory polyacrylamide-coated capillary, and only 0-6% was obtained in an uncoated fused-silica capillary. The RSD values for the intraday repeatability for an INSTCoated capillary were 0.1-1.0% (migration time) and 0.3-2.4% (peak area); RSD values for the interday repeatability in the same capillary are 0.6-1.4% (migration time) and 2.4-5.5% (peak area); RSD values for interday repeatability between different capillaries equaled 1.7-2.1% (migration time) and 2.8-10.9% (peak area). The INSTCoated capillary has been further used for rapid determination of globin chains isolated from red blood cells. A separation of α and β chains prepared from adult blood has been completed in 3 min with a peak resolution of 1.3, and the separation of α and (G)γ chains prepared from newborn blood took 3 min with a peak resolution of 3.6.
一种实验室自制的涂有INST的熔融石英毛细管最近被用于在pH值分别为3.0和2.5的磷酸钠背景电解质中对四种蛋白质混合物进行毛细管电泳分离。得到的分离效率范围为：肌红蛋白每米145,000理论塔板数至溶菌酶1,216,000 m⁻¹。在相同实验条件下，与涂有INST的毛细管相比，商用聚乙烯醇涂层毛细管获得的理论塔板数为49 - 89%，实验室聚丙烯酰胺涂层毛细管为0 - 86%，未涂层的熔融石英毛细管仅为0 - 6%。涂有INST的毛细管日内重复性的相对标准偏差（RSD）值为0.1 - 1.0%（迁移时间）和0.3 - 2.4%（峰面积）；同一毛细管日间重复性的RSD值为0.6 - 1.4%（迁移时间）和2.4 - 5.5%（峰面积）；不同毛细管之间日间重复性的RSD值为1.7 - 2.1%（迁移时间）和2.8 - 10.9%（峰面积）。涂有INST的毛细管已进一步用于快速测定从红细胞中分离出的珠蛋白链。从成人血液中制备的α链和β链在3分钟内完成分离，峰分辨率为1.3，从新生儿血液中制备的α链和(G)γ链的分离用时3分钟，峰分辨率为3.6。
