IF 3.8 2区 化学 Q1 BIOCHEMICAL RESEARCH METHODS
Jingwei Kou , Han Guo , Jing Leng , Houle Xiang , Hui Wang , Jinming Zhang , Pengpeng Yang , Fengxia Zou , Wei Zhuang , Huanqing Niu , Hanjie Ying , Jinglan Wu
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引用次数: 0

摘要

母乳中的低聚糖(HMOs)对婴儿的成长至关重要。最常用的 HMOs 分析方法是将亲水作用液相色谱法(HILIC)与超高效液相色谱法(UPLC)相结合。然而,高压操作和商用 HILIC 色谱柱的成本限制了 HMOs 研究的进展。在本研究中,我们通过将 2-丙烯酰胺基-2-甲基丙烷磺酸(AMPS)接枝到 Merrifield 树脂(PS-g-Cl)上,合成了一种新型混合模式吸附剂 PS-g-PAMPS,并对其进行了表征。将吸附剂装入色谱柱(混合-HILIC色谱柱),用于高效分析四种 HMO:2′-焦半乳糖(2′-FL)、N-四乳糖(LNnT)、3′-半乳糖(3′-SL)和 6′-半乳糖(6′-SL)。与AMINEX HPX-87H色谱柱、ROA-有机酸色谱柱和Glycan BEH酰胺色谱柱这三种商用色谱柱相比,我们的色谱柱具有几大优势:低压操作、吸附剂成本低、在相同的分析时间内实现六种分析物的基线分离。静电排斥和亲水相互作用的混合模式机制通过 zeta 电位和有机相比例得到了验证。评估了包括加标回收率、检出限和定量限在内的关键性能参数,并使用真实的 2′-FL 发酵样品确认了色谱柱的可靠性。这些结果证明了混合 HILIC 色谱柱在开发 HMO 应用色谱方面的潜力。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Preparation of mixed-mode hydrophilic particle for efficient separation of common human milk oligosaccharides
Human milk oligosaccharides (HMOs) are essential for babies’ growth. The most commonly used method for HMOs analysis combines hydrophilic interaction liquid chromatography (HILIC) with ultra-performance liquid chromatography (UPLC). However, high-pressure operation and the cost of commercial HILIC columns limit progress in the HMOs research. In this study, we synthesized a novel mixed-mode adsorbent, PS-g-PAMPS, by grafting 2-acrylamido-2-methylpropane sulfonic acid (AMPS) onto a Merrifield resin (PS-g-Cl), and characterized it. The adsorbents were packed into a column (Mixed-HILIC column) for efficient high-performance liquid chromatography (HPLC) analysis of four HMOs: 2′-Focusllactose (2′-FL), Lacto-N-Tetraose (LNnT), 3′-Sialyllactose (3′-SL), and 6′-Sialyllactose (6′-SL). Compared to three commercial columns, AMINEX HPX-87H column, ROA-Organic Acid column, Glycan BEH amide column, our column offered several advantages: low-pressure operation, cost-effective adsorbent, and baseline separation of six analytes in the same analysis time. The mixed-mode mechanism of electrostatic repulsion and hydrophilic interactions was verified using zeta potential, and organic phase ratio. Key performance parameters, including spike recovery, limit of detection, and limit of quantification, were evaluated, and the column's reliability was confirmed using a real 2′-FL fermentation sample. These results demonstrated the potential of the Mixed-HILIC column for the development of application chromatography in HMOs.
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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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