Chiari M, Micheletti C, Nesi M, Fazio M, Righetti P G
Instituto di Chimica degli Ormoni, CNR, Milano, Italy.
Electrophoresis. 1994 Feb;15(2):177-86. doi: 10.1002/elps.1150150131.
Matrices for electrokinetic separations, based on a unique class of mono- and disubstituted (on the amido nitrogen) acrylamides such as e.g., N-acryloylaminoethoxyethanol (AAEE) and acrylamido-N,N-diethoxyethanol, offer the following advantages: (i) strong resistance to alkaline hydrolysis (most zone separations occurring at basic pH values), (ii) high hydrophilicity and (iii) greater porosity, due to the higher molecular weight of the monomers. When compared with conventional poly(acrylamide), a poly(AAEE) matrix, when subjected to mild alkaline hydrolysis (0.1 N NaOH, 70 degrees C) appears to be 500 times more stable. Such stability is also confirmed under strong alkaline hydrolysis (1 N NaOH, 100 degrees C) as well as under mild and strong acidic hydrolysis. Mildly hydrolyzed poly(AAEE) matrices still perform extremely well in both conventional isoelectric focusing and immobilized pH gradients, techniques which are quite sensitive to traces of acrylate in the polymer coil. Conversely, mildly hydrolyzed poly(acrylamide) matrices, when used in isoelectric focusing, generate pH gradients between pH 4 and 5, having an inflection point (pH 4.6) equivalent to the pK value of acrylic acid. This novel class of monomers shows great promise for future applications in all electrokinetic methodologies.
基于一类独特的单取代和双取代(在酰胺氮上)丙烯酰胺(如N-丙烯酰基氨基乙氧基乙醇(AAEE)和丙烯酰胺基-N,N-二乙氧基乙醇)的电动分离基质具有以下优点:(i)对碱性水解具有很强的抗性(大多数区带分离在碱性pH值下进行),(ii)高亲水性,以及(iii)由于单体分子量较高而具有更大的孔隙率。与传统的聚丙烯酰胺相比,聚(AAEE)基质在进行温和的碱性水解(0.1N NaOH,70℃)时似乎比其稳定500倍。在强碱性水解(1N NaOH,100℃)以及温和和强酸性水解条件下,这种稳定性也得到了证实。轻度水解的聚(AAEE)基质在传统的等电聚焦和固定化pH梯度中仍然表现得非常出色,而这些技术对聚合物线圈中的微量丙烯酸酯非常敏感。相反,轻度水解的聚丙烯酰胺基质在等电聚焦中使用时,会在pH 4至5之间产生pH梯度,其拐点(pH 4.6)相当于丙烯酸的pK值。这类新型单体在未来所有电动方法中的应用前景广阔。