Determination of hydrogen peroxide using immobilized chemiluminescence reagents and flow injection analysis
Rao, Narasimha Murthy
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Permalink
https://hdl.handle.net/2142/20998
Description
Title
Determination of hydrogen peroxide using immobilized chemiluminescence reagents and flow injection analysis
Author(s)
Rao, Narasimha Murthy
Issue Date
1990
Doctoral Committee Chair(s)
Nieman, Timothy A.
Department of Study
Chemistry
Discipline
Chemistry
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Chemistry, Analytical
Language
eng
Abstract
In this work, use of immobilized luminol for the CL determination of hydrogen peroxide using flow injection analysis (FIA) was investigated. Nylon was initially chosen as the immobilization support because of its chemical inertness. Luminol was covalently immobilized on the inside of nylon tubing and used for FIA determination of peroxide using solution-phase hemin as catalyst (detection limit = 0.4 $\mu$M, relative standard deviation (rsd) = 2.5%). Next, horseradish peroxidase (HRP) was immobilized on nylon membranes. Luminol was immobilized on to an anion exchange resin and packed in a column. When used in the FIA mode, stoichiometric amounts of luminol could be released from the column by injecting another anion with higher affinity towards the resin. The pH downstream of the column could also be controlled by the amount of anion injected upstream of the column. Such columns were used in a single channel FIA scheme for the determination of peroxide in conjunction with immobilized HRP as catalyst (detection limit = 8 $\mu$M, mean rsd = 0.8%). The experiment was extended for the determination of glucose by using co-immobilized HRP and glucose oxidase (detection limit = 25 $\mu$M). The problem presented by real samples was solved using a sample pre-treatment technique.
The CL of luminol-Ru(bpy)$\sb3\sp{3+}$-peroxide system was investigated. It was found that CL is produced when the potential of a glassy carbon (GC) electrode coated with Nafion and loaded with Ru(bpy)$\sb3\sp{2+}$ is raised above 0.95 V in the presence of luminol and peroxide. The shape of the ECL vs applied potential curve obtained in this case was markedly different from that obtained using a bare GC electrode. The modified electrode (detection limit = 0.5 $\mu$M) as well as the bare GC electrode (detection limit = 0.2 $\mu$M) were used for peroxide determination at pH 8 in the presence of 10 $\mu$M luminol. In the presence of 1 mM peroxide, luminol could be detected down to 0.5 nM with the modified electrode. The modified electrode was also used for amperometric detection of peroxide (detection limit = 0.08 $\mu$M at pH 7.5).
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