Determination of pore size distribution of gel filtration media by low-field NMR

WANG Xiaojun; LIU Xinli; LI Xiunan; MA Guanghui; SU Zhiguo

doi:10.3969/j.issn.0438-1157.2013.11.028

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Determination of pore size distribution of gel filtration media by low-field NMR

更新时间：2025-12-10

- Determination of pore size distribution of gel filtration media by low-field NMR
- CIESC Journal Issue 11, Pages: 4090-4095(2013)
- 作者机构：
  
  1. 齐鲁工业大学食品与生物工程学院,山东,济南,250353
  2. 中国科学院过程工程研究所生化工程国家重点实验室,北京,100190
- 作者简介：
- 基金信息：
- DOI：10.3969/j.issn.0438-1157.2013.11.028
  CLC： Q028.5+3
- Online First：05 November 2013，
  
  Published：05 November 2013
- 稿件说明：
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WANG Xiaojun, LIU Xinli, LI Xiunan, et al. Determination of pore size distribution of gel filtration media by low-field NMR[J]. CIESC Journal, 2013, (11): 4090-4095.
DOI：

WANG Xiaojun, LIU Xinli, LI Xiunan, et al. Determination of pore size distribution of gel filtration media by low-field NMR[J]. CIESC Journal, 2013, (11): 4090-4095. DOI： 10.3969/j.issn.0438-1157.2013.11.028.

摘要

凝胶过滤介质在蛋白质、多糖等生物大分子的分离纯化过程中具有非常重要的作用

其中介质的孔径分布是决定分离纯化效果的关键因素。由于绝大部分凝胶过滤介质是软凝胶

因此很难用常规的方法如压汞法、低温氮吸附法等测定其孔径分布。本文探索了利用低场核磁共振测定凝胶过滤介质的孔径分布的方法。首先通过抽滤、自制琼脂糖凝胶块等实验确定了峰的归属

明确了介质孔内水、介质间隙水和自由水在核磁共振横向弛豫时间(T2)图谱上的分布范围;随后与逆体积排阻层析法(ISEC)测定的结果相对比

得出层析介质孔径和介质孔内水弛豫时间的关系;最后通过高斯正态拟合得到了介质的孔径分布。实验结果证实了低场核磁共振法测定凝胶过滤介质孔径分布的可行性。该法操作简单、测定迅速

并可以为其他层析介质孔径分布的测定提供参考。

Abstract

Gel filtration media play a very important role in separation and purification of bio-macromolecules such as proteins

polysaccharides.Their pore structure directly influences protein transport and adsorption in chromatographic separation column.The pore size distribution(PSD)plays a major role beyond simply the mean pore size.Various methods have been employed to determine the PSD of porous materials

including gas adsorption-desorption(BET)

mercury intrusion porosimetry(MIP).Since most of gel filtration media is soft gel

so it is difficult to measure their PSD by conventional method.In this paper

a new method

low-field NMR

was explored to determine the pore size distribution of gel filtration media.First

the distribution range of internal water

external water and free water in transverse relaxation time(T2)of low-field NMR were setting-out; then comparing the results with that from inverse size-exclusion chromatography(ISEC)and finding relationship between the pore size and relaxation time in internal water; finally

the pore size distribution of media was fitted by the Gaussian normal distribution.The results demonstrated that low-field NMR is an effective method for PSD determination of porous chromatographic media.The method is easy to operate

simple and quick

and can provide a reference for other determination of chromatography media pore size distribution.

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