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Luminol-Based Enhanced Chemiluminescence Assay for Quantification of Peroxidase and Hydrogen Peroxide in Aqueous Solutions: Effect of Reagent pH and Ionic Strength |
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| Authors: | Alok Bhandari Wongee Kim Keith Hohn |
| Affiliation: | 1Associate Professor, Dept. of Agricultural and Biosystems Engineering, Iowa State Univ., Ames, IA 50011 (corresponding author). E-mail: alokb@iastate.edu 2Graduate Student, Dept. of Civil Engineering, Kansas State Univ., Manhattan, KS 66501. 3Associate Professor, Dept. of Chemical Engineering, Kansas State Univ., Manhattan, KS 66501. |
| Abstract: | The effect of reagent pH and ionic strength was evaluated on the horseradish peroxidase (HRP)-luminol-p-iodophenol enhanced chemiluminescence assay. This assay was optimized for HRP and H2O2 quantification during enzyme-mediated remediation of groundwater or wastewater contaminated with phenolic chemicals. The maximum chemiluminescence intensity (ICLmax) and total area under the chemiluminescence intensity profiles were measured as a function of p-iodophenol concentration, reagent solution pH, and reagent ionic strength. ICLmax values were optimum at pH 8.5 and increased linearly with reagent ionic strength. Optimum chemiluminescence enhancement was produced at a p-iodophenol concentration of 0.3 mM under the reaction conditions. ICLmax values were linearly correlated with HRP and H2O2 concentrations within the ranges of 0.1–1 activity units/mL and 0.1–1 mM, respectively. Results indicate that the HRP-luminol-p-iodohenol enhanced chemiluminescence assay has a potential to be used for quantification of HRP activity and H2O2 concentration in aqueous solutions encountered in groundwater remediation or wastewater treatment scenarios. |
| Keywords: | Phenol Enzymes Hydrogen peroxide pH Wastewater management |