Artasensi, A.; Salina, E.; Fumagalli, L.; Regazzoni, L. A Novel Chromatographic Method to Assess the Binding Ability towards Dicarbonyls. Molecules2023, 28, 5341.
Artasensi, A.; Salina, E.; Fumagalli, L.; Regazzoni, L. A Novel Chromatographic Method to Assess the Binding Ability towards Dicarbonyls. Molecules 2023, 28, 5341.
Artasensi, A.; Salina, E.; Fumagalli, L.; Regazzoni, L. A Novel Chromatographic Method to Assess the Binding Ability towards Dicarbonyls. Molecules2023, 28, 5341.
Artasensi, A.; Salina, E.; Fumagalli, L.; Regazzoni, L. A Novel Chromatographic Method to Assess the Binding Ability towards Dicarbonyls. Molecules 2023, 28, 5341.
Abstract
Dicarbonyls are electrophiles able to generate the so-called advanced glycation end-products (AGEs). Human exposure occurs via ingestion (e.g., food), inhalation (e.g., electronic cigarettes) and dysregulation of endogenous metabolic pathways (e.g., glycolysis). Biological systems can prevent AGEs formation via enzymatic and nonenzymatic reactions. However, the continuous exposure to dicarbonyls may cause the depletion of endogenous substrates and the beginning of AGEs accumulation (i.e., carbonylation). Since this process has been associated with multiple disorders, several studies advocated the use of dicarbonyl binders as food preservatives or as drugs aimed at mitigating carbonylation damages. As reported for other toxicants (e.g., HNE, acrolein), the screening of selective and potent binders can be performed by measuring the residual amount of a selected dicarbonyl substrate, upon its incubation with candidate binders. Unfortunately, naturally occurring dicarbonyls are poorly responsive to many detectors (e.g., UV or MS). For this reason, most of the screening methods that have been reported to date relies on indirect quantification upon derivatization. However, this strategy can potentially lead to inaccurate results since the tested compounds can affect the derivatization yield. Therefore, we herein reported the use of benzylglyoxal (BGO) as a convenient, UV responsive, dicarboyl surrogate. By using BGO as a substrate, we setup an easy and cheap chromatographic assay for the direct measurement of the binding activity towards dicarbonyls, without derivatization. The method was qualified by assessing the binding ability of some known dicarbonyl binders, obtaining results consistent with literature.
Keywords
High performance liquid chromatography; binding assay; dicarbonyls; methylglyoxal; benzylglyoxal
Subject
Chemistry and Materials Science, Analytical Chemistry
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
