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首页> 外文期刊>Journal of chromatography, A: Including electrophoresis and other separation methods >Effect of flow behavior in extra-column volumes on the retention pattern of proteins in a small column
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Effect of flow behavior in extra-column volumes on the retention pattern of proteins in a small column

机译：小柱蛋白质保留模式的流动性能对额外柱体积的影响

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Experimental and theoretical analysis of deformation of band profiles in extra-column volumes (ECV) was performed, and its influence on the retention pattern of proteins in a small chromatographic column was quantified. Several macromolecule and small-molecule compounds, and their mixtures were eluted from a chromatographic system in the absence and presence of the column. The peak deformation in ECV was attributed to non-uniform velocity distribution in the radial direction in connecting capillaries. The phenomenon enhanced with increasing molecular weight of the model compound, when radial diffusion dominated the mechanism of band spreading. The band shape was also affected by the geometry of the injection system used, i.e., an injection loop capillary or a superloop. The phenomenon vanished for a small molecule compound, for which plug flow conditions could be established. The difference in flow behaviour of the macromolecule and small-molecule compounds caused them to migrate with different velocities in ECV, which resulted in partial separation of their bands. The ECV effect influenced the retention behaviour of macromolecules in a small column; it caused tailing of peaks and asymmetry of breakthrough curves. To describe the elution profiles in ECV and in the column, a mathematical model was used which accounted for non-ideality of the flow pattern. The model reproduced accurately band profiles of macromolecules within a range of relatively low velocities, typical however for protein chromatography. (C) 2019 Elsevier B.V. All rights reserved.
机译：进行了多柱体积（ECV）中带谱的变形的实验和理论分析，量化了对小色谱柱中蛋白质保留模式的影响。在不存在和存在的情况下，在色谱系统中洗脱几种大分子和小分子化合物，以及它们的混合物。 ECV中的峰值变形归因于连接毛细管中的径向上的不均匀速度分布。随着模型化合物的分子量增加，当径向扩散占主导地位时，随着模型化合物的分子量增强的现象。带状也受到使用的注射系统的几何形状的影响，即注射回路毛细管或超级棉。对于小分子化合物消失的现象，可以建立塞流条件。大分子和小分子化合物的流动行为的差异使得它们迁移在ECV中的不同速度，从而导致其带的部分分离。 ECV效应影响了小柱中大分子的保留行为;它引起了峰值和突破性曲线的不对称性。为了描述ECV和柱中的洗脱曲线，使用了一种数学模型，其考虑了流动模式的非理想性。该模型在相对低速度范围内再现大分子的准确带曲线，典型的蛋白质色谱。 （c）2019 Elsevier B.v.保留所有权利。

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