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Chemiluminescence examples
Written by Lluis Martinez | Thu, Dec 22, 2016

Chemiluminescence is the release of light from a chemical reaction. The

light is released either from a high-energy intermediate itself or when a high-
energy intermediate relaxes down to the lower energy final product. It is
important to remember that chemiluminesence is the result of a chemical
reaction, and is not the same as fluorescence. This means that
chemiluminescent reactions do not need an input of excitation light of any
kind. The overriding chemical formula of chemiluminescent reactions is A +
B→ C* + D → C + light + D

Where C* is the high energy intermediate.

Examples
Probably the most well-known chemiluminescence example i the reaction of
luciferin with ATP in the presence of the enzyme luciferase. This is the
reaction that occurs within fireflies to produce their light. This reaction could
be categorized as bioluminescence. An adaptation to this reaction was made
for use in the laboratory where peroxidase is used in place of luciferase as the
catalyzing enzyme. Enzyme protectors such as phenols, napthols, aromatic
amines, or benzothiazoles are added to the reaction to preserve the enzyme
and enhance the light output over several minutes. For this reason these
molecules are called “enhancers”.

Another chemiluminescence example is that of luminol with hydrogen

peroxide. This reaction ia catalyzed by the iron located in haemoglobin. It is
used at crime scenes to illuminate blood.

Luminol can also react with nitric oxide in air to produce an excited
aminophthalate anion intermediate that relaxes to produce water, molecular
nitrogen, and light. This reaction is used to detect nitric oxide levels in air to
determine pollution levels.

Another chemiluminescence example is the reaction of a peroxyoxalate such

as bis(2,4,6-trichlorophenyl)oxlate TCPO and hydrogen peroxide to produce a
high-energy intermediate 1,2-dioxetanedione. A fluorophore such as 9,10-
diphenylanthracene present in the solution will be excited by transfer of the
excited electron from the intermediate molecule and will emit light.

Two commonly used enzymes for chemiluminescence reactions are alkaline

phosphatase (AP) and horseradish peroxidase (HRP). These enzymes
catalyze oxidation of substrates in the presence of hydrogen peroxide
oxidizing agent. Whether the reaction produces a simple color change or light
is entirely dependent on the choice of substrate.

Another fun example of chemiluminescence can be observed in glow sticks.

When the stick is cracked it allows the reaction of the substrate cyalume plus
a dye with hydrogen peroxide to proceed to produce an excited dye that emits
light. The reaction has a limited lifetime, and the glow stick will eventually lose
its luminescence, but it lasts longer than most chemiluminescence reactions.
The dye releases light energy at a slower rate than most high-energy
intermediates. This idea can be transferred to the laboratory to extend the
lifetime of chemiluminescent reactions and to increase the time available for
imaging.

Chemiluminescent magnetic immunoassay

In a chemiluminescent magnetic immunoassay (CLIA) antibodies are labeled
with a chemiluminescent substrate. The corresponding antigen is bound to
paramagnetic beads. During an incubation period, the labeled antibodies bind
to the antigens on the magnetic beads. Then, the conjugated beads are
isolated from solution by magnetic separation. After isolation the reaction
solution is added. This reaction solution contains the other substrate and an
appropriate enzyme to produce a chemiluminescent reaction. Light is
produced only if the labeled antibodies matched the antigens on the bead
surface.

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