Fatima Saavedra

Luminol
Formal
Report
Section 10

Introduction
Chemiluminescence is a process where light is generated by a chemical reaction. This occurs
when the reaction produces a product in an excited electronic state that relaxes the ground
electronic state with concomitant emission of energy in the form of a photon-light. The excited
state may relax to the ground state through radiationless (loss of energy through emission of
visible light). This can also be seen when fluorescence and phosphorescence occur (involve
singlet and triplet electronic states).
A number of living organisms emit light through chemiluminescene reactions (they involve
naturally occurring compounds). Bioluminescence is an example of chemiluminescence in which
a biochemical process results in the emission of light. An example of this would be fireflies.
When they are in search of a mate, the male will emit flashes of visible light that result from the
reaction of luciferin with molecular oxygen in a process catalyzed by the enzyme luciferase
(Reaction 1).

Reaction 1
The light that is produced by fireflies and other bioluminescent organisms has baffled observers
for years. There are many organisms that have developed the ability to emit light. They include
jellyfish, corals, squids, fishes, insects, and marine worms. Other than fish, this list doesnt
include higher forms of life. The chemiluminescent compound in fireflies is called luciferin. The
enzyme is the one that induces it to emit the light-which is called luciferase.
The chemiluminescent properties of luminol have a number of practical applications. A forensic
assay that used to identify the presence of blood at a crime scene is one of these. When the
alkaline solution of luminol and hydrogen peroxide is exposed to blood, then the characteristic of

the blueis-green light associated with the chemiluminescence of luminol is observed. The iron
catalyst is required for the reaction is provided by the hemoglobin present in blood.
Data and Observation
1.0236 grams of 3-nitrophthalic acid was used.
Some of the product got stuck on the stirring rod.
Diluted stock solution caused a blue luminescence. When the stock was added, the luminescence
increased. The bright color faded out to a died out purple.
Discussion
Luminol is synthesized by a sequence of reactions that involves the condensation of 3nitrophthalic acid with hydrazine to produce 3-nitrophthalhydrazide, which is then reduced by
sodium dithionite to form luminol (Reaction 2). The chemiluminescent reaction is demonstrated
by mixing luminol with hydrogen peroxide and potassium ferricyanide (Reaction 3).

Reaction 2

Reaction 3
The formation of 3-Nitrophthalhydrazide is basically a reaction in which 1,2-dicarboxylic acid
and 1,2-diamine react to form a cyclic diamide or hydrazide (Reaction 4). The acid and amine
are in rapid equilibrium with the corresponding ammonium salt and the carboxylate ion (the
latter is not subject to a nucleophilic attack at the carbonyl carbon atom). The acid may undergo
nucleophilic attack; however, the small amount of it that exists in equilibrium reacts with the free
amine to produce the tetrahedral intermediate.

Reaction 4

The reaction was purified by crystallization and vacuum filtration. This purification takes
advantage of the fact that the impurities often have a different solubility than the compound that
is being purified. Acetic acid was used in the reaction instead of HCl because acetic acid is a
weak acid. Acetic acid will weakly ionize, which would lead to fewer H+ ions, while HCl would
ionize strongly.
Conclusion
The expected blue fluorescence was observed upon the combination of the solutions for the
luminal preparation. This would mean that the synthesis occurred correctly. Luminol is used in
forensic investigations in order to trace blood that could have been left behind at crime scenes.
Luminol reacts with the iron that is found in the hemoglobin. Luminol can be sprayed across an
area, and the trace amounts of the activating oxidant will cause the lumiol to emit a blue glow
that can be seen in a dark room. The glow doesnt last long, but it can be captured with a
photograph. Sources of error that could have contributed to the experiment would be unclean
glassware, incorrect measurements of products, and mixing the wrong compounds.
Reference
Experimental Organic Chemistry 10th Edition