Alizarin Red S as an electrochemical indicator for saccharide recognition

In addition to the well-established spectroscopic Alizarin Red S (ARS) assay for the determination of binding constants between arylboronic acids and different saccharides, we report the use of ARS as a reporter in an electrochemical set-up. The electrochemical properties of ARS, the binding to phenyl boronic acid (PBA) and the competition with fructose in phosphate buffer at pH 7.4 were investigated by cyclic voltammetry (CV). By choosing a negative scan direction (starting at +0.2 V), a quasi-reversible process was detected at E0′ = −0.59V with ΔEp = 0.1V. An irreversible oxidation peak at +0.42 V could also be detected. These peaks are characterised both as a 2-proton-2-electron transfer and corresponds to the oxidation and reduction of the anthraquinone or the ortho-quinone moiety. After addition of phenylboronic acid a new oxidation peaks occurred at −0.42 V which correlates with the ARS–PBA interaction. The peak current increased with increasing phenylboronic acid concentration according to the release of BA and formation of the ARS–PBA ester. After addition of fructose the peak current decreases again, in proportion to the fructose concentration, enabling the use of ARS as an electrochemical reporter for fructose detection up to 50 mM. Also the interaction with other cis-diol containing compounds such as sorbitol, mannitol, glucose and mannose was investigated and a dependence based on already published binding constants to phenylboronic acid could be shown.

Graphical abstractFigure optionsDownload full-size imageDownload as PowerPoint slideHighlights► Transfer of the established Alizarin Red S (ARS) for saccharide–boronic acid interaction to electrochemistry. ► Investigation of electrochemical behaviour of ARS and its interaction with phenylboronic acid at pH 7.4. ► Through addition of fructose the ARS was displaced. ► Displaced ARS could be monitored electrochemically corresponding to the used fructose concentration.