Abstract
The purpose of this study was to examine the application of the coordinated cis-[Cr(C2O4)(pm)(OH)2]+ cation where pm denotes pyridoxamine, as a specific sensing ion for the detection of hydrogen peroxide (H2O2). The proposed method for H2O2 detection includes two key steps. The first step is based on the nonenzymatic decarboxylation of pyruvate upon reaction with H2O2, while the second step is based on the interaction of cis-[Cr(C2O4)(pm)(OH2)2]+ with the CO2 released in the previous step. Using this method H2O2 generated during glutamate-induced oxidative stress was detected in HT22 hippocampal cells. The coordination ion cis-[Cr(C2O4)(pm)(OH2)2]+ and the spectrophotometric stopped-flow technique were applied to determine the CO2 concentration in cell lysates, supernatants and cell-free culture medium. Prior to CO2 assessment pyruvate was added to all samples studied. Pyruvate reacts with H2O2 with 1:1 stoichiometry, and consequently the amount of CO2 released in this reaction is equivalent to the amount of H2O2.
Highlights
Reactive oxygen species (ROS) generation contributes to the ethiology of many diseases, including diabetes, arteriosclerosis or neurodegenerative disorders and others [1,2,3]
The cis-[Cr(C2O4)(pm)(OH2)2]+ ion was previously developed as a specific molecular biosensor to detect uptake of CO2, generated in the reaction between Na2CO3 and HCl [20]
During the carbon dioxide uptake by cis-[Cr(C2O4)(pm)(OH2)2]+ complex ion the most significant changes of absorbance were seen at λ = 560 nm
Summary
Reactive oxygen species (ROS) generation contributes to the ethiology of many diseases, including diabetes, arteriosclerosis or neurodegenerative disorders and others [1,2,3]. The increase in intracellular production of free radicals may lead to cellular damage, including alterations of lipids, proteins and DNA [4]. ROS [5,6], in particular superoxide anion (O2−), hydroxyl radical (OH) or hydrogen peroxide (H2O2) and reactive nitrogen species [7,8,9,10], e.g., nitrogen dioxide, are highly cytotoxic. H2O2 which in turn can be transformed into highly reactive and toxic hydroxyl radicals. H2O2 is one of the most important mediators of oxidative stress detected under pathological conditions. H2O2 is probably involved in the neuronal damage seen in Parkinson’s and Huntington’s diseases and other neuronal disorders
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