User:Helen L. Slucher/Notebook/CHEM 571/2013/10/01

Project name

Main project page Previous entry      Next entry

Objective Monitor the kinetics and yield of the horseradish peroxidase-catalyzed oxidation of luminol. Description UV-Vis Absorbance of reactants, catalysts, and products horseradish peroxidase (Use stock solution) luminol (use stock solution) 3-aminophthalic acid (product of reaction between luminol and H2O2 catalyzed by HRP. In order to take this measurement, react (in 1:1 ratio or with slight excess H2O2) luminol with H2O2 in presence of HRP. Allow the reaction to proceed for 5 minutes; take the spectrum) Kinetics of luminol oxidized by changes in absorption spectrum, reaction carried out in stopped flow mixer Add HRP/Luminol stock solution to stopped flow mixer Add H2O2 stock solution to stopped flow mixer equilibrate mixer tubes with sample. Initiate Mixing Using luminol and 3-aminophthalic acid spectra as endpoints, determine the kinetics of 3-aminophthalic acid synthesis. Data Series 1: HRP Stock Series 2: 0.126mM Luminol Series 3: H2O2 Stock Notes Stock Buffer = 5.1 mM HRP = 0.77 uM Luminol = 1.51 mM H2O2 = 45 mM Luminol dilution solution: 0.126 mM