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Simultaneous Measurement of Superoxide/Hydrogen Peroxide and NADH Production by Flavin-containing Mitochondrial Dehydrogenases
Published on: February 24, 2018
A rapid and economical method for the quantification of hydrogen peroxide (H2O2) using a modified HPLC apparatus
Omar Tantawi1, Abbas Baalbaki1, Rime El Asmar1
1American University of Beirut, Faculty of Arts and Sciences, Department of Chemistry, P.O. Box 11-0236, Riad El Solh, 1107-2020 Beirut, Lebanon.
Abstract:
H2O2 is one of the most commonly used oxidants for the degradation of recalcitrant organic contaminants in advanced oxidation processes (AOPs). However, most research aiming to optimize AOPs is missing the monitoring of the remaining H2O2, an important parameter to assess the efficiency of the process. In this work, a novel method for [H2O2] quantification was developed using simple modifications of an HPLC-DAD setup that is available in most analytical chemistry laboratories. The modifications include the use of acidified potassium iodide solution as mobile phase and replacing the reverse phase column with a series of capillary columns. This instrument configuration allowed also the quantification of organic contaminants using the same H2O2 containing sample. The method's LOD and LOQ were calculated to be as low as 8.29 × 10-4 mM and 2.76 × 10-3 mM, respectively with an LDR range of 0.01-150 mM. The cost per analysis ranged between 0.8 and 1.8 USD cents depending on the concentration tested. This analytical method was validated by a statistical comparison to a well-known titrimetric method that is commonly used for H2O2 quantification. It was also tested using standards prepared in natural matrices such as spring and seawater, and in media containing high concentration of several spectator species such as chlorides, bicarbonates, humic acids, fumaric acids and micro pollutants. The method showed excellent robustness by maintaining high regression coefficient and excellent sensitivity in all calibration curves regardless of the matrix content.
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