生物大分子液相色谱分离中色谱柱长与柱径比的优化研究

Studies on Optimization of Ratio of Column Length to Its Diameter for the Separation of Biopolymers by Liquid Chromatography

用厚度为 1cm、而直径不同的色谱饼 (或饼形色谱柱 )对 7种蛋白质进行了色谱谱分离 ,得出了柱长与柱径比对生物大分子分离度影响不大的结论。还对柱内腔几何体积相同、而柱长 柱径比不同的色谱柱和色谱饼对分离度的影响以及它们压力降作了比较 ,结果表明色谱饼可在几乎不降低分离度的前提下显著降低色谱系统的压力。通过对 1 0× 5 0mmI.D .色谱饼体积和质量负载量以及质量回收率的测定 ,表明色谱饼对生物大分子的分离已达到制备规模。

The separation of seven proteins was carried out on the chromatographic pie (pie-like columns) with 1. 0 cm in thickness and different diameters. It is concluded that the ratios of column length to the diameter have little effect on the separation of biopolymers. Also, the different influences of a 200 x 7.9 nun I. D. column and a 5 x 50 min I. D. chromatographic pie with the same interior volume for both on resolution and the differential pressure between these two types of column pie have been compared with each other, the results indicate that the chromatographic pie can significantly lower the hack pressure with little decrease in resolution. The injection amount to a 5 x 50 nun I. D. chromatographic pie and mass recovery determined show that the chromatographic pie can he used for the separation of biopolymers on a preparative scale.