Mixed-mode hydrophilic interaction/ion-exchange liquid chromatography ? Separation potential in peptide analysis

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216208%3A11310%2F21%3A10435850" target="_blank" >RIV/00216208:11310/21:10435850 - isvavai.cz</a>

Výsledek na webu

<a href="https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=FV_SqRq.UN" target="_blank" >https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=FV_SqRq.UN</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.microc.2021.106158" target="_blank" >10.1016/j.microc.2021.106158</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Mixed-mode hydrophilic interaction/ion-exchange liquid chromatography ? Separation potential in peptide analysis

Popis výsledku v původním jazyce

In this work, three recently developed HILIC columns, i.e., HILIC-A containing silica gel, HILIC-N containing polyhydroxy phase and HILIC-B containing aminopropyl, were characterized in detail to compare their peptide retention, selectivity, and peak symmetry. The detailed study of chromatographic behavior of peptides revealed that a multimodal retention mechanism is present in systems with these stationary phases. Using three model analytes (caffeine, benzenesulfonic acid, and benzyltrimethylammonium chloride), we demonstrated that the HILIC-A column behaves as a HILIC/cation exchanger, and the HILIC-B column as a HILIC/anion exchanger depending on the mobile phase pH and buffer concentration. Furthermore, the pH (ammonium formate, pH 2.1 and 3.5; ammonium acetate pH 5.5) and buffer concentration (10?50 mM) of the mobile phase affect peptide retention and peak symmetry. Satisfactory retention and peak shape of peptides can be obtained by balancing ionic and hydrophilic properties of both analyte and stationary phases. Therefore, buffer concentration and pH optimization is an effective tool for manipulating retention and peak symmetry, thereby enhancing the potential of these columns in the analysis of diverse compounds. The potential application of these columns in the separation of biologically active peptides was confirmed by achieving baseline separation under optimized gradient elution.

Název v anglickém jazyce

Mixed-mode hydrophilic interaction/ion-exchange liquid chromatography ? Separation potential in peptide analysis

Popis výsledku anglicky

In this work, three recently developed HILIC columns, i.e., HILIC-A containing silica gel, HILIC-N containing polyhydroxy phase and HILIC-B containing aminopropyl, were characterized in detail to compare their peptide retention, selectivity, and peak symmetry. The detailed study of chromatographic behavior of peptides revealed that a multimodal retention mechanism is present in systems with these stationary phases. Using three model analytes (caffeine, benzenesulfonic acid, and benzyltrimethylammonium chloride), we demonstrated that the HILIC-A column behaves as a HILIC/cation exchanger, and the HILIC-B column as a HILIC/anion exchanger depending on the mobile phase pH and buffer concentration. Furthermore, the pH (ammonium formate, pH 2.1 and 3.5; ammonium acetate pH 5.5) and buffer concentration (10?50 mM) of the mobile phase affect peptide retention and peak symmetry. Satisfactory retention and peak shape of peptides can be obtained by balancing ionic and hydrophilic properties of both analyte and stationary phases. Therefore, buffer concentration and pH optimization is an effective tool for manipulating retention and peak symmetry, thereby enhancing the potential of these columns in the analysis of diverse compounds. The potential application of these columns in the separation of biologically active peptides was confirmed by achieving baseline separation under optimized gradient elution.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10406 - Analytical chemistry

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2021

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Microchemical Journal

ISSN

0026-265X

e-ISSN

—

Svazek periodika

165

Číslo periodika v rámci svazku

June

Stát vydavatele periodika

NL - Nizozemsko

Počet stran výsledku

8

Strana od-do

106158

Kód UT WoS článku

000647789500009

EID výsledku v databázi Scopus

2-s2.0-85103124991