Multifaceted separations approach for elucidation of the physical and chemical properties of extracellular hydrocolloids

IF 3.8 2区 化学 Q1 BIOCHEMICAL RESEARCH METHODS
Kaitlin C. Lesco , Stefanie Van Wychen , Arnav Deshpande , Lieve M.L. Laurens , S. Kim Ratanathanawongs Williams
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引用次数: 0

Abstract

A multifaceted separations platform that incorporates the strengths of asymmetrical flow field-flow fractionation with multi-detectors (AF4-MD), high performance anion exchange chromatography (HPAEC), and hydrophilic interaction liquid chromatography with mass spectrometry (HILIC-MS) is developed to obtain a more complete picture of the molecular weights (MW), composition, and salt-induced aggregation behavior of extracellular polymeric substances (EPS) produced by the algae Chlorella vulgaris. The absence of a stationary phase makes AF4-MD particularly well suited for characterizing polydisperse hydrocolloid polymers as well as studies that investigate the effect of ionic environments that aligns with the natural environment of C. vulgaris. Fractionation of C. vulgaris EPS revealed three distinct MW populations ranging from 4 × 10⁴ to 3 × 10⁸ Daltons. This exceeds the previously reported MW by three orders of magnitude and reports a previously unknown size subpopulation. The optimized AF4-MD technique was then used to produce two size fractions that were probed using HPAEC and LC-MS. These orthogonal methods uncovered compositional heterogeneity across fractions, with variations in monosaccharides and amino acids.
AF4-MD is also well suited for studying the behavior of EPS in the presence of different salts. For each salt studied, e.g., NaNO3, NaCl, and MgCl2, an increase in solution ionic strength results in aggregation as corroborated by a shift to higher MWs. Each salt exhibited distinct effects on EPS aggregation, with NaCl causing the least aggregation and MgCl2 the most. These findings highlight the need for multiple techniques when analyzing complex polymers such as EPS and the benefits of AF4-MD in elucidating complex polymer behaviors in different ionic environments.
细胞外水胶体的物理和化学性质的阐明的多方面分离方法
为了更全面地了解普通小球藻(Chlorella vulgaris)胞外聚合物(EPS)的分子量(MW)、组成和盐诱导聚集行为,建立了一个多方面的分离平台,该平台结合了非对称流场-多检测器流动分离(AF4-MD)、高性能阴离子交换色谱(HPAEC)和亲水相互作用液相色谱-质谱(HILIC-MS)的优势。固定相的缺失使得AF4-MD特别适合于表征多分散的水胶体聚合物,以及研究与C. vulgaris自然环境相一致的离子环境的影响。对寻常草EPS进行分离,发现3个不同的分子量居群,范围在4 × 10⁴至3 × 10⁸道尔顿之间。这超过了先前报道的MW三个数量级,并报告了一个以前未知的大小亚群。然后利用优化后的AF4-MD技术制备两种大小的组分,分别用HPAEC和LC-MS进行检测。这些正交方法揭示了组分的异质性,在单糖和氨基酸的变化。AF4-MD也非常适合于研究EPS在不同盐存在下的行为。对于所研究的每种盐,例如NaNO3、NaCl和MgCl2,溶液离子强度的增加导致聚集,这可以通过向更高分子量的转变得到证实。不同盐对EPS的聚集作用不同,其中NaCl的聚集作用最小,MgCl2的聚集作用最大。这些发现强调了在分析EPS等复杂聚合物时需要多种技术,以及AF4-MD在阐明不同离子环境下复杂聚合物行为方面的优势。
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来源期刊
Journal of Chromatography A
Journal of Chromatography A 化学-分析化学
CiteScore
7.90
自引率
14.60%
发文量
742
审稿时长
45 days
期刊介绍: The Journal of Chromatography A provides a forum for the publication of original research and critical reviews on all aspects of fundamental and applied separation science. The scope of the journal includes chromatography and related techniques, electromigration techniques (e.g. electrophoresis, electrochromatography), hyphenated and other multi-dimensional techniques, sample preparation, and detection methods such as mass spectrometry. Contributions consist mainly of research papers dealing with the theory of separation methods, instrumental developments and analytical and preparative applications of general interest.
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