The Chemiluminescence Of Luminol Biology Essay

Chemiluminiscence is the procedure whereby visible radiation is produced via a chemical reaction with the development of small or no heat. Chemiluminescence, unlike phosphorescence and fluorescence, does non necessitate to absorb an external beginning of photons to make an aroused province. Alternatively chemiluminescence occurs when a molecule is excited through a series of chemical reactions. The molecule, normally in the lower aroused vest province, can spontaneously lose its electronic energy in the signifier of a photon. In fluorescence the electronic spin ( a?†S=0 ) is preserved, while the beaming strength of chemiluminescence can be expressed as:

This equation holds true since the rate of chemiluminescence depends both on the quantum output C?CL, and besides the rate of chemical reaction, . Therefore one may observe that the grade of chemiluminescence depends greatly on the quantum output and the figure of photons emitted per chemiluminescencing molecule.

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In the practical below luminol, a chemiluminescence molecule is to be prepared from 3-nitrophthalic acid. Luminol is seen to hold many applications in life. For illustration, male fire beetles in hunt for a mate are able to bring forth certain chemicals to let their lower venters to glow. The interaction of luciferin from the fire beetle, its enzyme luciferase together with adenosine triphosphate, O and the Mg ion allows for this natural chemiluminescence. First the luciferin is adenylated by the ATP:

Luciferin + ATP & A ; agrave ; Luciferyl Adenylate + PP

The 2nd reaction involved the merchandise of the first reaction being oxidised to give decarboxylketoluciferin. This is seen to be a cyclic endoperoxide intermediate that consequences in an electronically aroused decarboxylketoluciferin, that eventually chemilumineses.

Luciferyl Adenylate + O2 & A ; agrave ; decarboxylketoluciferin* + AMP +CO2

Decarboxylketoluciferin* & A ; agrave ; decarboxylketoluciferin + photon

The decomposition of the endoperoxide consequences in an aroused carbonyl that leads to chemiluminescence.


2.1 ) Chemicals



Trade name

3-nitrophthalic acid

98 % Pure


Hydrazine 8 % aqueous solution





Hopkin and Williams

Sodium hydrated oxide



Sodium dithionite

Lab Reagent Grade

Fisher Scientific

Glacial acetic acid



Potassium Ferricyanide


Riedel de Hean

3 % Hydrogen peroxide



2.2 ) Apparatus

Analytic balance Tripod

Weighing boat Gauze

Spatula vacuity pump

Filter tubing ( dimensions 20 X 150 millimeter ) Filter funnel

10 milliliter mensurating cylinder Hirsh funnel

100 milliliter mensurating cylinder Filter paper

Stiring rod conelike flask with side arm

Thermometer trial tubing ( 20 c 150 millimeters )

Rubber bung 50 milliliter beaker

Bunsen burner 100 milliliter beaker

Erlenmeyer flask stirring rod

2.3 ) Procedure

A ) Preparation

1.000g of 3-nitrophthalic acid were weighed on an analytical balance and placed in a filter tubing of approximative dimensions of 20 ten 150 millimeter.

2 milliliter of 8 % v/v aqueous solution of hydrazine was added with attention.

This solution was gently heated over a Bunsen fire until the solid dissolved. ( The trial tubing placed about 20 centimeters off from the fire and sporadically traveling it off from the fire )

3 milliliter of Digol was added and stoppered utilizing a gum elastic spile fitted with a thermometer.

The side arm was connected to the vacuity pump and boiled to disill off extra H2O in the solution

The solution was heated to 110-130 OC so quickly heated to 200 OC.

The reaction mixture was kept at this temperature for two proceedingss.

The mixture was so cooled to 100 OC and 15 milliliter of hot H2O was added

The solution as cooled under running H2O and the intermediate merchandise ( II ) collected by suction filtration utilizing a Hirsch funnel.

The nitro-compound was transferred to a 20 ten 150 mm trial tubing

5 milliliter of 10 % Na hydrated oxide and 3.000 g of Na dithionite were added

Any staying solids on the sides of the trial tubing was washed with a minimal sum of distilled H2O

The mixture was boiled gently for 5 proceedingss while stirring

2 milliliter of glacial acetic acid was added to the mixture and the solution cooled under tap H2O.

The aminoalkane ( III ) was collected by suction filtration utilizing a Hirsh funnel

B ) Chemiluminescence Demonstration ( carried out in the dark )

2 milliliter of 10 % Na hydroxide solution was assorted with 18 milliliters of H2O

The 0.013 g of aminoalkane ( III ) was added to this solution and stirred until wholly dissolved. ( stock solution A )

The oxidizing solution ( stock solution B ) was prepared by blending 4 milliliter of 3 % w/v solution K ferricyanide to an equal volume of 3 % H peroxide.

The solution as made up to 200 milliliter utilizing distilled H2O

Chemiluminescence was observed by thining 5 milliliter of stock solution A to 40 milliliters distilled H2O.

Stock solution B was poured at the same time with the solution prepared into a funnel placed on a big Erlenmeyer flask

The flask was swirled and little parts of base were added to increase the glare.

2.4 ) Precautions

Care was taken when utilizing the Hydrazine due to its toxicity

The 3-nitrophthalic acerb solution was heated with periodic remotion from the fire.

Care was taken to reassign all the solid from the filter paper, without adding any filter paper to the solid.

The chemiluminescence presentation was carried out in a dark room so for better observation of light

2.5 ) Beginnings Of mistake

Some losingss due to transportations may hold been present taking to a lower output

Some filter paper may hold been transferred with the aminoalkane therefore taking to a higher output.

3. Consequences

Mass of aminoalkane ( III ) produced= 0.13 g

Observation- Blue fluorescence was observed upon combination of the solutions as described above

4. Discussion

Nitrophthalic acid was added to hydrazine, and with the presence of heat the nitro derived function, Luminol, is formed. This may be described as a desiccation reaction.

Luminol may so organize the Dianion by oxidization utilizing the H peroxide, Na hydrated oxide, and the usage of the K ferricyanide as the accelerator. The dianion is oxidised to the three excited province so due to a slow intercrossing system is converted to a vest dianion excited province. This compound finally decays to its land province and emits a photon of light per molecule.

The energy released in the reaction is hence seen to do the excitement of negatrons in the merchandises of the reaction therefore the exited negatrons return to the land province, and bluish seeable visible radiation is emitted. Since blood is seen to hold the metal ion Fe2+ , it may be used to oxidize the H peroxide into O and H2O. Thus the O may be used to oxidize the luminol in the same manner as seen above. Therefore blood can be seen to luminate with the add-on of luminol

The output of the aminoalkane produced was seen to be rather low. This may be caused by the overall entropically unfavorable reaction. This therefore would do low reaction efficiency. The reaction above is seen to necessitate back-to-back intermolecular and intramolecular condensations, which are likely to hold, and entropy less than nothing. Therefore with the net negative information and high temperatures, the reaction is seen to be unfavorable.


From this practical one may see that a bright bluish luminescence was observed. This therefore confirms the production of luminol. The comparatively low output was due to the fact that the reaction was entropically unfavorable.


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