Validation of a High-Performance Liquid Chromatography Separation of Weak Acids, Neutral Compounds, and Permanent Ions Using Sequential Elution with Tandem-Column Liquid Chromatography

IF 2.8 3区工程技术 Q2 CHEMISTRY, ANALYTICAL

Journal of separation science Pub Date : 2025-03-20 DOI:10.1002/jssc.70124

Lauren Kline Lovejoy, Joe P. Foley, Kaitlin M. Grinias

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Abstract

Sequential elution-liquid chromatography (SE-LC) employs conventional high-performance LC instrumentation to separate compounds by class and within each class using two or more sequential elution modes. The aim of this work was to validate a novel SE-LC method used to separate weak acids, permanently charged ions (anions), and neutral compounds by applying first a low pH isocratic elution to elute the weak acids, then an acetonitrile gradient to elute the neutral compounds, and lastly a sodium methanesulfonate gradient to elute the anionic compounds using a tandem-column arrangement comprised of a C18 column and a strong anion exchange column. The effects of flow rate, starting mobile phase organic content, column temperature, mobile phase composition, and gradient start time were investigated using a 2^5-1 fractional factorial design with center points for robustness. The method was demonstrated to be robust, specific, linear, accurate, and precise (repeatable) over the range of 6%–120% of the target analyte concentrations.

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串联柱液相色谱连续洗脱对弱酸、中性化合物和永久离子的高效液相色谱分离验证。

顺序洗脱-液相色谱（SE-LC）采用传统的高效液相色谱仪器，通过两种或更多的顺序洗脱模式，按类别和每个类别分离化合物。这项工作的目的是验证一种新的SE-LC方法，该方法用于分离弱酸，永久带电离子（阴离子）和中性化合物，首先使用低pH等容洗脱来洗脱弱酸，然后使用乙腈梯度来洗脱中性化合物，最后使用由C18柱和强阴离子交换柱组成的串联柱排列，使用甲磺酸钠梯度来洗脱阴离子化合物。采用25-1分数因子设计，以中心点为稳健性，考察了流速、起始流动相有机含量、柱温、起始流动相组成和梯度起始时间的影响。结果表明，该方法在目标分析物浓度的6%-120%范围内具有鲁棒性、特异性、线性、准确和精确（可重复）的特点。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of separation science 化学-分析化学

CiteScore

6.30

自引率

16.10%

发文量

408

审稿时长

1.8 months

期刊介绍： The Journal of Separation Science (JSS) is the most comprehensive source in separation science, since it covers all areas of chromatographic and electrophoretic separation methods in theory and practice, both in the analytical and in the preparative mode, solid phase extraction, sample preparation, and related techniques. Manuscripts on methodological or instrumental developments, including detection aspects, in particular mass spectrometry, as well as on innovative applications will also be published. Manuscripts on hyphenation, automation, and miniaturization are particularly welcome. Pre- and post-separation facets of a total analysis may be covered as well as the underlying logic of the development or application of a method.