The effect of hydrodynamics on chemiluminescence (CL)
emission is investigated for the
tris(2,2‘-bipyridine)ruthenium(II/III)
(Ru(bpy)3
2+/3+)
electrogenerated chemiluminescence (ECL) system in solution phase and immobilized in Nafion. The maximum CL emission is obtained
at low linear velocities for slow reactions, at
intermediate
linear velocities for reactions with intermediate
kinetics,
and at high linear velocities for fast reactions.
Relative
kinetic information for a variety of analytes is acquired
by comparing charge-coupled device (CCD) images to
stopped-flow and flow injection analysis data obtained
with
a photomultiplier tube (PMT). CCD images are also
utilized to observe reproducibility and homogeneity of
different Nafion film preparations. The spatial
resolution
provided by the CCD allows a more comprehensive
analysis of the effects of flow on observed CL reaction
kinetics than is possible with a PMT. These studies
yield
important guidelines for application and optimization of
CL detection in flow injection, liquid chromatography, and
capillary electrophoresis.
The stopped-flow technique was employed to measure chemiluminescent emission from the reaction of a mixture of oxalate and proline with a chemiluminescence reagent, tris(2,2'-bipyridine) ruthenium(III), or Ru(bpy)3(3+) is a versatile reagent and is often used in bioanalytical applications, including the detection of certain drugs and their metabolites, for example. Unfortunately, Ru(bpy)3(3+) has not yet been fully examined as a possible chemiluminescence reagent for simultaneous kinetic determinations. In this work, a differential reaction rate method, based on simple least squares regressions of the pseudo-first order decay data, was used to resolve two compounds, oxalate and proline, reacting simultaneously with Ru(bpy)3(3+). Our results indicate that stopped-flow analyses with Ru(bpy)3(3+) could provide a viable method for simultaneous determinations of unresolvable analytes of environmental and pharmaceutical importance.
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