Spectrophotometric determination of hydrogen peroxide in water with peroxidase-catalyzed oxidation of potassium iodide and its applications to hydroxylamine-involved Fenton and Fenton-like systems

A spectrophotometric method for the rapid measurement of hydrogen peroxide (H₂O₂) in aqueous solutions was developed in this study. This method is based on a reaction catalyzed by peroxidase (POD) in which potassium iodide (KI) is oxidized to generate the stable yellow-colored I₃^- within 15 s. The absorbance of the generated I₃^- at both 350 nm and 400 nm had good linear relationships with H₂O₂ concentration in the range of 0-70 μM (R² > 0.999) with sensitivities of 2.34 × 10⁴ M^-1 cm^-1 and 5.30 × 10³ M^-1 cm^-1 respectively. Meanwhile, through calculation, the detection limits of the proposed POD-KI method at 350 nm and 400 nm were 0.09 μM and 0.33 μM, respectively. Even when the concentration of H₂O₂ was up to 350 μM, the absorbance of the generated I₃^- at 350 nm did not decrease observably. The generated I₃^- was found to be stable enough in ultrapure water, underground water, reservoir water and samples containing the strong reducing agent hydroxylamine. Moreover, the proposed POD-KI method was successfully used to analyze trace H₂O₂ in rainwater, and to monitor the change of H₂O₂ concentration in the Fenton, hydroxylamine/Fenton and hydroxylamine/Cu(II)/H₂O₂ systems. Overall, the POD-KI method could be adopted as a candidate method to determine H₂O₂ in Fenton and Fenton-like systems, and especially in those involving hydroxylamine.