Date of Award

Spring 1981

Project Type


Program or Major


Degree Name

Doctor of Philosophy


A new approach to analysis with chemiluminescent detection is proposed and demonstrated. Two Phase Chemiluminescence utilizes an immobilized reagent bulk phase and a sample phase which contains the analyte. The analyte is permitted to diffuse into the reagent phase under controlled conditions. A chemiluminescent reaction occurs in the reagent phase, which is quantitated and related to the analyte concentration in the sample phase. A general theory is presented to describe the relationship between analyte concentration and chemiluminescent intensity.

The reaction between luminol and hydrogen peroxide was used in a probe configuration. The reaction was catalyzed by peroxidase immobilized on one end of the fiber optic. A non-linear response for hydrogen peroxide was found at a pH of 9 and 10('-3) M luminol concentration. The detection limit for peroxidase was found to be 10('-6) M.

The reaction between adenosine triphosphate (ATP) and firefly luciferine was monitored in a flow cell configuration. The firefly luciferase was immobilized in the presence of luciferin. A detection limit for ATP under nonoptimized conditions was estimated at 2x10('-8) M ATP. With the current design, a calibration curve for ATP concentration could not be constructed due to the decay of the signal intensity with exposure of the reagent phase to ATP.

The chemiluminescence of a peramino ethylene; 1,1',3,3', tetraethyl (DELTA)('2,2')bi(imidazolidine) (EIA); was used to develop an oxygen sensor. A 10% EIA solution in the hexane was immobilized behind a Teflon membrane. Oxygen diffusing through the membrane reacted with the EIA producing a signal proportional to the oxygen partial pressure in both the gaseous and aqueous phases. The detection limit for oxygen in the gas phase was estimated to be about 1 ppm(V/V).