Gwarda Radosław Ł, Aletańska-Kozak Monika, Matosiuk Dariusz, Dzido Tadeusz H
Department of Physical Chemistry, Chair of Chemistry, Faculty of Pharmacy with Medical Analytics Division, Medical University of Lublin, 4a Chodźki St., 20-093 Lublin, Poland.
Chair and Department of Synthesis and Chemical Technology of Pharmaceutical Substances, Faculty of Pharmacy with Medical Analytics Division, Medical University of Lublin, 4a Chodźki St., 20-093 Lublin, Poland.
J Chromatogr A. 2016 Apr 1;1440:240-248. doi: 10.1016/j.chroma.2016.02.064. Epub 2016 Feb 24.
Our previous results show, that C18 silica-based adsorbents used in high-performance thin-layer chromatography (HPTLC), provide complex retention mechanism basing on various polar and nonpolar interactions. Here we present, that in chromatography of peptides, due to mixed-mode properties of these adsorbents, there is a simple way to obtain inversion of separation system type (from reversed-phase, RP, to normal-phase, NP, and vice versa). The results presented provide detailed information how to obtain inversion mentioned and reflect the extent (the type and concentration of organic solvent, the type and concentration of ion-pairing reagent in the mobile phase) of this phenomenon. We show, that the system type inversion results in significant change of selectivity of separation, which may be especially useful in 2D separation of complex samples of basic/amphoteric compounds such as peptides. This results from the fact, that C18 silica-based HPTLC adsorbents, may be used in hydrophilic interaction chromatography (HILIC) or RP chromatography, in dependence on mobile phase composition.
我们之前的结果表明，用于高效薄层色谱（HPTLC）的基于C18硅胶的吸附剂，基于各种极性和非极性相互作用提供了复杂的保留机制。在此我们展示，在肽的色谱分析中，由于这些吸附剂的混合模式特性，有一种简单的方法可以实现分离系统类型的反转（从反相，RP，到正相，NP，反之亦然）。所呈现的结果提供了关于如何实现上述反转的详细信息，并反映了这种现象的程度（流动相中有机溶剂的类型和浓度、离子对试剂的类型和浓度）。我们表明，系统类型的反转会导致分离选择性的显著变化，这在二维分离诸如肽等碱性/两性化合物的复杂样品时可能特别有用。这是因为基于C18硅胶的HPTLC吸附剂，根据流动相组成，可用于亲水相互作用色谱（HILIC）或RP色谱。
