Preparation of linear polyacrylamide-coated capillaries - Study of the polymerization process and its effect on capillary electrophoresis performance
Preparation of linear polyacrylamide-coated capillaries - Study of the polymerization process and its effect on capillary electrophoresis performance
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DOI：
10.1016/s0021-9673(98)00923-6
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发表时间：
1999-01-15
影响因子：
4.1
通讯作者：
Diez-Masa, JC
中科院分区：
文献类型：
--
作者：
Cifuentes, A;Canalejas, P;Diez-Masa, JC
研究了不同参数对熔融石英毛细管内壁线性聚丙烯酰胺涂层特性的影响及其对毛细管电泳性能的影响。为了进行这项研究，一个可重复的程序，以获得与7-辛-1-烯基三甲氧基硅烷的表面硅烷醇的改性程度相似的毛细管首先接近。接下来，研究了聚合物通过与接枝到二氧化硅上的甲硅烷基试剂的共价键连接到二氧化硅壁上。以这种方式，通过使用具有类似硅烷化程度的柱，在不同条件下涂覆的毛细管之间观察到的CE性能的差异可以归因于聚丙烯酰胺层的特性。它表明，这些聚合物涂层的特性和再现性取决于聚合的温度和所使用的聚合溶解的脱气的充分控制。更有趣的是，它也表明，单体（丙烯酰胺），引发剂（过硫酸铵）和活化剂（N，N，N ′，N ′-四甲基乙二胺）的量，以及它们之间的比例用于制备涂层聚合物对CE柱的性能有很大的影响。讨论了制备聚丙烯酰胺涂料的最佳条件。证明了这些线性聚丙烯酰胺涂层毛细管在自由区CE中分离碱性和酸性蛋白质的适用性。此外，这些涂层柱在毛细管凝胶电泳分离DNA片段的使用。(C)1999 Elsevier Science B. V.保留所有权利。
The effect of different parameters controlling the characteristics of linear polyacrylamide coatings deposited on the inner wall of fused-silica capillaries and their influence on capillary electrophoresis (CE) performance of these coated columns is investigated. To carry out this study, a reproducible procedure to obtain capillaries with similar extent of modification of the surface silanols with 7-oct-1-enyltrimethoxisilane was first approached. Next the polymer attachment to the silica wall, via covalent linkage to the silyl reagent grafted onto the silica, was investigated. In this way, by using columns with a similar silylation extent, differences in CE performance observed among capillaries coated under diverse conditions could be assigned to the characteristics of the polyacrylamide layer. It is demonstrated that the characteristics and reproducibility of these polymeric coatings depend on the adequate control of both the temperature of polymerization and the degassing of the polymerizing dissolutions used. More interestingly, it is also demonstrated that the quantities of monomer (acrylamide), initiator (ammonium persulfate) and activator (N,N,N',N'-tetramethylethylenediamine), and the ratio among them used in the preparation of the coating polymer have a large influence on the performance of CE columns. The optimum conditions for preparing the polyacrylamide coatings are discussed. The applicability of these linear polyacrylamide-coated capillaries to the separation of basic and acidic proteins in free zone CE is demonstrated. Besides, the use of these coated columns in capillary gel electrophoresis for the separation of DNA fragments is shown. (C) 1999 Elsevier Science B.V. All rights reserved.