High Pressure Size Exclusion Chromatography (HPSEC) of humic substances: molecular sizes, analytical parameters, and column performance
High Pressure Size Exclusion Chromatography (HPSEC) of humic substances: molecular sizes, analytical parameters, and column performance
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腐殖质的高压尺寸排阻色谱 (HPSEC)：分子尺寸、分析参数和色谱柱性能
DOI：
10.1016/s0045-6535(98)00198-2
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发表时间：
1999
期刊：
影响因子：
8.8
通讯作者：
Alessandro Piccolo
中科院分区：
文献类型：
--
作者：
P. Conte;Alessandro Piccolo
高压粒径排除色谱法（HPSEC）越来越多地用于评估不同来源腐殖质的分子大小。以已知分子量的多糖为标准，对两种不同的HPSEC色谱柱（TSK G3000SW和Biosep S2000）进行不对称因子（As）、理论板数(N)、分布系数（kd）和柱分辨率（Rs）的测定。两柱的校准曲线相同，而分析参数显示TSK柱在分离多糖标准物方面的效率仅略高。腐殖质的MWand - mnvalue根据每个色谱柱的分子量范围不同而不同，但两种色谱柱的相对标准偏差从未超过5%。柱间的变化归因于固有的腐殖质特性，如构象结构的稳定性。这些结果表明，溶液中的腐殖质物质是小分子的松散结合，这些小分子可能通过尺寸排除孔的扩散而持续分散。HPSEC被证实是一种高度精确的方法来评估溶解腐殖质的相对分子大小分布。
High Pressure Size Exclusion chromatography (HPSEC) is increasingly used to evaluate molecular sizes of humic substances from different sources. Asymmetry factors (As), number of theoretical plates (N), coefficient of distribution (kd), and column resolution (Rs) were determined for two different HPSEC columns (TSK G3000SW and Biosep S2000) and polysaccharides of known molecular weights were used as standards. Calibration curves were equivalent for both columns whereas analytical parameters revealed that the TSK column was only slightly more efficient in separating polysaccharide standards. MWand Mnvalues for humic substances differed according to the molecular weight range of each column but relative standard deviation never exceeded 5% for both columns. Variations between columns were attributed to intrinsic humic properties such as the stability of conformational structures. These results suggested that humic substances in solutions are loosely-bound association of small molecules that may be consistently dispersed by diffusion through size-exclusion pores. HPSEC is confirmed to represent a highly precise method to evaluate the relative molecular-size distribution of dissolved humic substances.