Abstract
Despite the importance of hydrogen peroxide (H2O2) in initiating oxidative damage and its connection to various diseases, the detection of low concentrations of H2O2 (<10 μM) is still limited using current methods, particularly in non-aqueous systems. One of the most common methods is based on examining the color change of a reducing substrate upon oxidation using UV/Vis spectrophotometry, fluorophotometry and/or paper test strips. In this study, we show that this method encounters low efficiency and sensitivity for detection of ultratrace amounts of H2O2 in non-aqueous media. Thus, we have developed a simple, fast, accurate and inexpensive method based on UV/Vis spectrophotometry to detect H2O2 in non-aqueous systems, such as alcohols. In this regard, we demonstrate that monitoring the Soret and Q-band regions of high-valent iron-oxo (ferryl heme) intermediates in horseradish peroxidase (HRP) is well suited to detect ultratrace amounts of H2O2 impurities in alcohols in the range of 0.001–1000 μM using UV/Vis spectrophotometry. We monitor the optical spectra of HRP solution for the red shift in the Soret and Q-band regions upon the addition of alcohols with H2O2 impurity. We also monitor the reversibility of this shift to the original wavelength over time to check the spontaneous decay of ferryl intermediates to the ferric state. Thus, we have found that the ferryl intermediates of HRP can be used for the detection of H2O2 in alcohols at μg L−1 levels through via UV/Vis spectrophotometric method.
Highlights
Hydrogen peroxide (H2O2) is an essential oxygen metabolite in living systems and serves as a messenger in cellular signal transduction.[1]
Common hydrogen peroxide test strips were used to detect H2O2 impurity in a 100% ethanol sample, the minimum detectability of these strips is reported as 30 mM H2O2 (Fig. 1a(i))
Has a Soret of 418 nm and two Q-bands of 527 nm and 557 nm (Fig. 1b).[43,44,45]. We hypothesized that these indicative peaks could be used to detect the H2O2 impurity of alcohols and developed a simple spectrophotometric method utilizing horseradish peroxidase (HRP) solutions to detect and quantify H2O2 based on the formation of ferryl intermediates (Fig. 1c).[44]
Summary
Hydrogen peroxide (H2O2) is an essential oxygen metabolite in living systems and serves as a messenger in cellular signal transduction.[1]. Has a Soret of 418 nm and two Q-bands of 527 nm and 557 nm (Fig. 1b).[43,44,45] We hypothesized that these indicative peaks could be used to detect the H2O2 impurity of alcohols and developed a simple spectrophotometric method utilizing HRP solutions to detect and quantify H2O2 based on the formation of ferryl intermediates (Fig. 1c).[44]
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