An Enzyme-based Spectrophotometric Method for the Determination of Phenolic Compound (2- Methoxyphenol) Using Peroxidase from Ipomea batata

Abstract

Phenolic compounds and their derivatives are considered priority pollutants because they are harmful to living organisms even at low concentrations. Due to their toxicity and persistence in the environment, many efforts are been made to develop simple and effective methods for their determination. This study describes a peroxidase-based method for the quantitative determination of a named phenolic compound. The method is based on the oxido-reductase activity of peroxidase in a H2O2/2-Methoxyphenol system. Calibration curve of peroxidase activity (absorbance) against the phenol concentration forms the basis for its quantitative estimation. Factors influencing the reaction were evaluated and optimized; Optimum pH and temperature were 6 and 40°C respectively while optimum reaction time was 6 minutes.  The calibration curve for the analyte was linear (R2 =0.998) within the concentration range of 0.01–5 mM.  Repeatability of analysis was 3.8% RSD for 7 replicate measurements. Recovery tests for the analyte in water samples gave values between 85.33 – 112%.