Lemque R, Vidal-Madjar C, Racine M, Piquion J, Sébille B
Laboratoire de Physico-Chimie des Biopolymères, CNRS, Université Paris-Val-de-Marne, Thiais, France.
J Chromatogr. 1991 Aug 16;553(1-2):165-77. doi: 10.1016/s0021-9673(01)88485-5.
The anion-exchange elution behaviour of alpha-lactalbumin was studied on cross-linked and quaternized polyvinylimidazole, deposited on various high-performance liquid chromatographic supports (porous silica and diol silica). The influence of the nature and thickness of the coating layer on the retention and band-width properties of the protein elution peak was examined by isocratic elution. The retention properties of alpha-lactalbumin were studied from the plot of log k' vs. log([NaCl]), where k' is the capacity factor and [NaCl] the displacer salt concentration in the aqueous phase. The retention depends on the amount of stationary phase deposited on the support, but an increased hydrophobic effect is found when the polymer films do not coat the chromatographic support uniformly. Band broadening of the elution peaks was studied in terms of plots of plate height vs. mobile phase velocity. An important mass-transfer contribution is found, which decreases with increasing k' and increases with the thickness of the coating layer. These effects reveal that the diffusion into the polymer layer is the controlling step of the ion-exchange process with non-uniform polymer layers of large mean thickness.
研究了α-乳白蛋白在交联和季铵化聚乙烯咪唑上的阴离子交换洗脱行为，该聚合物沉积在各种高效液相色谱载体（多孔硅胶和二醇硅胶）上。通过等度洗脱考察了涂层的性质和厚度对蛋白质洗脱峰保留和带宽性质的影响。从log k'对log([NaCl])的图中研究了α-乳白蛋白的保留性质，其中k'是容量因子，[NaCl]是水相中置换盐的浓度。保留取决于沉积在载体上的固定相的量，但当聚合物膜不能均匀地覆盖色谱载体时，会发现疏水作用增强。根据塔板高度对流动相速度的图研究了洗脱峰的峰展宽。发现了一个重要的传质贡献，它随着k'的增加而减小，随着涂层厚度的增加而增加。这些效应表明，对于平均厚度较大的非均匀聚合物层，向聚合物层的扩散是离子交换过程的控制步骤。
