Simon József, Kovács Márton, Szabó Pál T
MS Metabolomics Research Laboratory, Centre for Structural Science, Hungarian Research Network (HUN-REN), Magyar tudósok körútja 2, H-1117 Budapest, Hungary.
Research Group of Analytical Chemistry, University of Pannonia, Egyetem utca 10, H-8200 Veszprém, Hungary.
Heliyon. 2025 Feb 5;11(4):e42461. doi: 10.1016/j.heliyon.2025.e42461. eCollection 2025 Feb 28.
The chromatographic measurement of polar molecules is often surprisingly complicated. On the one hand, the standard experiments use C18 columns, which is usually unsuitable. On the other hand, these types of molecules can behave vastly differently despite their similarities. Thus, finding the right chromatographic conditions is challenging. HILIC can be an obvious choice, but C18 is still used with suitable analytes. Comparing the two methods would be advantageous, but a numerical comparison can be problematic. Thus, a simple comparing and ranking system is needed and put to use to find the best method for the separation of polar basic molecules.
In this paper, nine columns with different stationary phases, from reversed-phase to hydrophilic interaction, are compared and ranked in gradient elution. The effect of pH was also considered. The measurements were repeated in acidic, near-to-neutral, and basic conditions. A straightforward system is developed to rank different stationary phases. Its foundation was peak shapes and resolutions. Every measurement condition with all the columns on the available pHs is evaluated by each property. The testing solution consisted of ten components, which aimed to cover a wide range of polar basic molecules. The comparison also focuses on these analytes. We highlighted which analyte is adaptable to different methods or which needed exclusive conditions. The resulting best column with the optimal conditions is presented and proven highly efficient for their separation.
A straightforward comparison and ranking system is developed to test multiple chromatographic columns with different stationary phases, from reversed-phase to hydrophilic interaction in gradient elution, seeking the best method for separating polar basic molecules. Nine columns were compared in multiple conditions. The best setup resulting in the competition is presented in detail, which can be applied to a wide range of analytes.
极性分子的色谱测量往往出奇地复杂。一方面，标准实验使用C18柱，这通常并不合适。另一方面，尽管这些类型的分子具有相似性，但它们的行为可能有很大差异。因此，找到合适的色谱条件具有挑战性。亲水作用色谱法（HILIC）可能是一个明显的选择，但C18柱仍用于合适的分析物。比较这两种方法会很有优势，但数值比较可能会有问题。因此，需要一个简单的比较和排名系统并加以应用，以找到分离极性碱性分子的最佳方法。
本文比较了九种具有不同固定相的色谱柱，从反相到亲水相互作用，并在梯度洗脱中进行了排名。还考虑了pH值的影响。在酸性、近中性和碱性条件下重复进行了测量。开发了一个简单的系统来对不同的固定相进行排名。其基础是峰形和分离度。根据每种特性评估在可用pH值下使用所有色谱柱的每个测量条件。测试溶液由十种成分组成，旨在涵盖广泛的极性碱性分子。比较也集中在这些分析物上。我们强调了哪些分析物适用于不同的方法，或者哪些需要特殊条件。给出了在最佳条件下得到的最佳色谱柱，并证明其对分离非常有效。
开发了一个简单的比较和排名系统，用于测试多种具有不同固定相的色谱柱，从反相到亲水相互作用，在梯度洗脱中寻找分离极性碱性分子的最佳方法。在多种条件下比较了九种色谱柱。详细介绍了竞争中产生的最佳设置，该设置可应用于广泛的分析物。
