Chemiluminescence

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A chemoluminescent reaction in an Erlenmeyer flask Chemoluminescent reaction.jpg
A chemoluminescent reaction in an Erlenmeyer flask

Chemiluminescence (also chemoluminescence) is the emission of light (luminescence) as the result of a chemical reaction, i.e. a chemical reaction results in a flash or glow of light. A standard example of chemiluminescence in the laboratory setting is the luminol test. Here, blood is indicated by luminescence upon contact with iron in hemoglobin. When chemiluminescence takes place in living organisms, the phenomenon is called bioluminescence. A light stick emits light by chemiluminescence.

Contents



Physical description

As in many chemical reactions, chemiluminescence starts with the combining of two compounds, say A and B, to give a product C. Unlike most chemical reactions, the product C converts to a further product, which is produced in an electronically excited state often indicated with an asterisk:

A + B → C
C → D*

D* then emits a photon (hν), to give the ground state of D: [1] I

D* → D + hν

In theory, one photon of light should be given off for each molecule of reactant. In practice, the yield ("quantum efficiency") is often low owing to side reactions.

For example, A could be luminol and [B] hydrogen peroxide. D would be 3-APA (3-aminophthalate).

Chemiluminescence differs from fluorescence or phosphorescence in that the electronic excited state is the product of a chemical reaction rather than of the absorption of a photon. It is the antithesis of a photochemical reaction, in which light is used to drive an endothermic chemical reaction. Here, light is generated from a chemically exothermic reaction. The chemiluminescence might be also induced by an electrochemical stimulus, in this case is called electrochemiluminescence.

Bioluminescence in nature: A male firefly mating with a female of the species Lampyris noctiluca. Lampyris Noctiluca (firefly) mating.gif
Bioluminescence in nature: A male firefly mating with a female of the species Lampyris noctiluca .

Liquid-phase reactions

Chemiluminescence was first observed with lophine (triphenylimidazole). [2] When in basic solution, this compound converts to the imidazolate, which reacts with oxygen to eventually give a dioxetane. Fragmentation of the dioxetane gives the excited state of an anionic diamide. [3]

Steps leading up to chemiluminescence of lophine. LophineMech.svg
Steps leading up to chemiluminescence of lophine.

Chemiluminescence in aqueous system is mainly caused by redox reactions. [4]

Chemiluminescence after a reaction of hydrogen peroxide and luminol Chemiluminescence.jpg
Chemiluminescence after a reaction of hydrogen peroxide and luminol

Gas-phase reactions

Green and blue glow sticks Knicklichter.jpg
Green and blue glow sticks
The activated NO2[] luminesces broadband visible to infrared light as it reverts to a lower energy state. A photomultiplier and associated electronics counts the photons that are proportional to the amount of NO present. To determine the amount of nitrogen dioxide, NO2, in a sample (containing no NO) it must first be converted to nitric oxide, NO, by passing the sample through a converter before the above ozone activation reaction is applied. The ozone reaction produces a photon count proportional to NO that is proportional to NO2 before it was converted to NO. In the case of a mixed sample that contains both NO and NO2, the above reaction yields the amount of NO and NO2 combined in the air sample, assuming that the sample is passed through the converter. If the mixed sample is not passed through the converter, the ozone reaction produces activated NO2[] only in proportion to the NO in the sample. The NO2 in the sample is not activated by the ozone reaction. Though unactivated NO2 is present with the activated NO2[], photons are emitted only by the activated species that is proportional to original NO. Final step: Subtract NO from (NO + NO2) to yield NO2 [8]

Infrared chemiluminescence

In chemical kinetics, infrared chemiluminiscence (IRCL) refers to the emission of infrared photons from vibrationally excited product molecules immediately after their formation. The intensities of infrared emission lines from vibrationally excited molecules are used to measure the populations of vibrational states of product molecules. [9] [10]

The observation of IRCL was developed as a kinetic technique by John Polanyi, who used it to study the attractive or repulsive nature of the potential energy surface for gas-phase reactions. In general the IRCL is much more intense for reactions with an attractive surface, indicating that this type of surface leads to energy deposition in vibrational excitation. In contrast reactions with a repulsive potential energy surface lead to little IRCL, indicating that the energy is primarily deposited as translational energy. [11]

Enhanced chemiluminescence

Enhanced chemiluminescence (ECL) is a common technique for a variety of detection assays in biology. A horseradish peroxidase enzyme (HRP) is tethered to an antibody that specifically recognizes the molecule of interest. This enzyme complex then catalyzes the conversion of the enhanced chemiluminescent substrate into a sensitized reagent in the vicinity of the molecule of interest, which on further oxidation by hydrogen peroxide, produces a triplet (excited) carbonyl, which emits light when it decays to the singlet carbonyl. Enhanced chemiluminescence allows detection of minute quantities of a biomolecule. Proteins can be detected down to femtomole quantities, [12] [13] well below the detection limit for most assay systems.

Applications

Biological applications

Chemiluminescence has been applied by forensic scientists to solve crimes. In this case, they use luminol and hydrogen peroxide. The iron from the blood acts as a catalyst and reacts with the luminol and hydrogen peroxide to produce blue light for about 30 seconds. Because only a small amount of iron is required for chemiluminescence, trace amounts of blood are sufficient.

In biomedical research, the protein that gives fireflies their glow and its co-factor, luciferin, are used to produce red light through the consumption of ATP. This reaction is used in many applications, including the effectiveness of cancer drugs that choke off a tumor's blood supply[ citation needed ]. This form of bioluminescence imaging allows scientists to test drugs in the pre-clinical stages cheaply. Another protein, aequorin, found in certain jellyfish, produces blue light in the presence of calcium. It can be used in molecular biology to assess calcium levels in cells. What these biological reactions have in common is their use of adenosine triphosphate (ATP) as an energy source. Though the structure of the molecules that produce luminescence is different for each species, they are given the generic name of luciferin. Firefly luciferin can be oxidized to produce an excited complex. Once it falls back down to a ground state a photon is released. It is very similar to the reaction with luminol.

Many organisms have evolved to produce light in a range of colors. At the molecular level, the difference in color arises from the degree of conjugation of the molecule, when an electron drops down from the excited state to the ground state. Deep sea organisms have evolved to produce light to lure and catch prey, as camouflage, or to attract others. Some bacteria even use bioluminescence to communicate. The common colors for the light emitted by these animals are blue and green because they have shorter wavelengths than red and can transmit more easily in water.

In April 2020, researchers reported having genetically engineered plants glow much brighter than previously possible by inserting genes of the bioluminescent mushroom Neonothopanus nambi . The glow is self-sustained, works by converting plants' caffeic acid into luciferin and, unlike for bacterial bioluminescence genes used earlier, has a relatively high light output that is visible to the naked eye. [20] [21] [22] [23]

Chemiluminescence is different from fluorescence. Hence, fluorescent proteins such as Green fluorescent protein are not chemiluminescent. However, combining GFP with luciferases allows bioluminescence resonance energy transfer (BRET), which increases the quantum yield of light emitted in these systems.

See also

Related Research Articles

Biophotons are photons of light in the ultraviolet and low visible light range that are produced by a biological system. They are non-thermal in origin, and the emission of biophotons is technically a type of bioluminescence, though bioluminescence is generally reserved for higher luminance luciferin/luciferase systems. The term biophoton used in this narrow sense should not be confused with the broader field of biophotonics, which studies the general interaction of light with biological systems.

<span class="mw-page-title-main">Bioluminescence</span> Emission of light by a living organism

Bioluminescence is the production and emission of light by living organisms. It is a form of chemiluminescence. Bioluminescence occurs widely in marine vertebrates and invertebrates, as well as in some fungi, microorganisms including some bioluminescent bacteria, and terrestrial arthropods such as fireflies. In some animals, the light is bacteriogenic, produced by symbiotic bacteria such as those from the genus Vibrio; in others, it is autogenic, produced by the animals themselves.

<span class="mw-page-title-main">Nitric oxide</span> Colorless gas with the formula NO

Nitric oxide is a colorless gas with the formula NO. It is one of the principal oxides of nitrogen. Nitric oxide is a free radical: it has an unpaired electron, which is sometimes denoted by a dot in its chemical formula. Nitric oxide is also a heteronuclear diatomic molecule, a class of molecules whose study spawned early modern theories of chemical bonding.

<span class="mw-page-title-main">Glow stick</span> Self-contained, short-term light source

A glow stick, also known as a light stick, chem light, light wand, light rod, and rave light, is a self-contained, short-term light-source. It consists of a translucent plastic tube containing isolated substances that, when combined, make light through chemiluminescence. The light cannot be turned off and can be used only once. The used tube is then thrown away. Glow sticks are often used for recreation, such as for events, camping, outdoor exploration, and concerts. Glow sticks are also used for light in military and emergency services applications. Industrial uses include marine, transportation, and mining.

<span class="mw-page-title-main">Luciferase</span> Enzyme family

Luciferase is a generic term for the class of oxidative enzymes that produce bioluminescence, and is usually distinguished from a photoprotein. The name was first used by Raphaël Dubois who invented the words luciferin and luciferase, for the substrate and enzyme, respectively. Both words are derived from the Latin word lucifer, meaning "lightbearer", which in turn is derived from the Latin words for "light" (lux) and "to bring or carry" (ferre).

<span class="mw-page-title-main">Phosphorescence</span> Process in which energy absorbed by a substance is released relatively slowly in the form of light

Phosphorescence is a type of photoluminescence related to fluorescence. When exposed to light (radiation) of a shorter wavelength, a phosphorescent substance will glow, absorbing the light and reemitting it at a longer wavelength. Unlike fluorescence, a phosphorescent material does not immediately reemit the radiation it absorbs. Instead, a phosphorescent material absorbs some of the radiation energy and reemits it for a much longer time after the radiation source is removed.

<span class="mw-page-title-main">Luminol</span> Chemical compound

Luminol (C8H7N3O2) is a chemical that exhibits chemiluminescence, with a blue glow, when mixed with an appropriate oxidizing agent. Luminol is a white-to-pale-yellow crystalline solid that is soluble in most polar organic solvents, but insoluble in water.

<span class="mw-page-title-main">Luciferin</span> Class of light-emitting chemical compounds

Luciferin is a generic term for the light-emitting compound found in organisms that generate bioluminescence. Luciferins typically undergo an enzyme-catalyzed reaction with molecular oxygen. The resulting transformation, which usually involves breaking off a molecular fragment, produces an excited state intermediate that emits light upon decaying to its ground state. The term may refer to molecules that are substrates for both luciferases and photoproteins.

Photodissociation, photolysis, photodecomposition, or photofragmentation is a chemical reaction in which molecules of a chemical compound are broken down by photons. It is defined as the interaction of one or more photons with one target molecule.

<span class="mw-page-title-main">Diphenyl oxalate</span> Chemical compound

Diphenyl oxalate is a solid whose oxidation products are responsible for the chemiluminescence in a glowstick. This chemical is the double ester of phenol with oxalic acid. Upon reaction with hydrogen peroxide, 1,2-dioxetanedione is formed, along with release of the two phenols. The dioxetanedione then reacts with a dye molecule, decomposing to form carbon dioxide and leaving the dye in an excited state. As the dye relaxes back to its unexcited state, it releases a photon of visible light.

<span class="mw-page-title-main">Emmett Chappelle</span> American scientist (1925-2019)

Emmett W. Chappelle was an American scientist who made valuable contributions in the fields of medicine, philanthropy, food science, and astrochemistry. His achievements led to his induction into the National Inventors Hall of Fame for his work on bioluminescence, in 2007. Being honored as one of the 100 most distinguished African American scientists of the 20th Century, he was also one of the members of the American Chemical Society, the American Society of Biochemistry and Molecular Biology, the American Society of Photobiology, the American Society of Microbiology, and the American Society of Black Chemists.

<span class="mw-page-title-main">Firefly luciferase</span>

Firefly luciferase is the light-emitting enzyme responsible for the bioluminescence of fireflies and click beetles. The enzyme catalyses the oxidation of firefly luciferin, requiring oxygen and ATP. Because of the requirement of ATP, firefly luciferases have been used extensively in biotechnology.

<span class="mw-page-title-main">Firefly luciferin</span> Chemical compound

Firefly luciferin is the luciferin, or light-emitting compound, used for the firefly (Lampyridae), railroad worm (Phengodidae), starworm (Rhagophthalmidae), and click-beetle (Pyrophorini) bioluminescent systems. It is the substrate of luciferase, which is responsible for the characteristic yellow light emission from many firefly species.

A photocyte is a cell that specializes in catalyzing enzymes to produce light (bioluminescence). Photocytes typically occur in select layers of epithelial tissue, functioning singly or in a group, or as part of a larger apparatus. They contain special structures termed as photocyte granules. These specialized cells are found in a range of multicellular animals including ctenophora, coelenterates (cnidaria), annelids, arthropoda and fishes. Although some fungi are bioluminescent, they do not have such specialized cells.

<span class="mw-page-title-main">Horseradish peroxidase</span> Chemical compound and enzyme

The enzyme horseradish peroxidase (HRP), found in the roots of horseradish, is used extensively in biochemistry applications. It is a metalloenzyme with many isoforms, of which the most studied type is C. It catalyzes the oxidation of various organic substrates by hydrogen peroxide.

<span class="mw-page-title-main">Renilla-luciferin 2-monooxygenase</span>

Renilla-luciferin 2-monooxygenase, Renilla luciferase, or RLuc, is a bioluminescent enzyme found in Renilla reniformis, belonging to a group of coelenterazine luciferases. Of this group of enzymes, the luciferase from Renilla reniformis has been the most extensively studied, and due to its bioluminescence requiring only molecular oxygen, has a wide range of applications, with uses as a reporter gene probe in cell culture, in vivo imaging, and various other areas of biological research. Recently, chimeras of RLuc have been developed and demonstrated to be the brightest luminescent proteins to date, and have proved effective in both noninvasive single-cell and whole body imaging.

<span class="mw-page-title-main">Electrochemiluminescence</span> Emission of light from electrochemical reactions

Electrochemiluminescence or electrogenerated chemiluminescence (ECL) is a kind of luminescence produced during electrochemical reactions in solutions. In electrogenerated chemiluminescence, electrochemically generated intermediates undergo a highly exergonic reaction to produce an electronically excited state that then emits light upon relaxation to a lower-level state. This wavelength of the emitted photon of light corresponds to the energy gap between these two states. ECL excitation can be caused by energetic electron transfer (redox) reactions of electrogenerated species. Such luminescence excitation is a form of chemiluminescence where one/all reactants are produced electrochemically on the electrodes.

<span class="mw-page-title-main">Peroxyoxalate</span>

Peroxyoxalates are esters initially formed by the reaction of hydrogen peroxide with oxalate diesters or oxalyl chloride, with or without base, although the reaction is much faster with base:

Photoelectrochemical processes are processes in photoelectrochemistry; they usually involve transforming light into other forms of energy. These processes apply to photochemistry, optically pumped lasers, sensitized solar cells, luminescence, and photochromism.

Chemiluminescence is the emission of light through a chemical reaction. It contrasts with fluorescence, which is excited by a light source. During chemiluminescence, the vibrationally excited product of an exoergic chemical reaction relaxes to its ground state with the emission of photons. Since the process does not require excitation light, problems in its application caused by light scattering or source instability are absent, and there is no concern about autofluorescence in the background, which can lead to highly sensitive deep tissue imaging.

References

  1. Vacher, Morgane; Fdez. Galván, Ignacio; Ding, Bo-Wen; Schramm, Stefan; Berraud-Pache, Romain; Naumov, Panče; Ferré, Nicolas; Liu, Ya-Jun; Navizet, Isabelle; Roca-Sanjuán, Daniel; Baader, Wilhelm J.; Lindh, Roland (March 2018). "Chemi- and Bioluminescence of Cyclic Peroxides". Chemical Reviews. 118 (15): 6927–6974. doi:10.1021/acs.chemrev.7b00649. PMID   29493234.
  2. Radziszewski, B. R. (1877). "Untersuchungen über Hydrobenzamid, Amarin und Lophin". Berichte der Deutschen Chemischen Gesellschaft (in German). 10 (1): 70–75. doi:10.1002/cber.18770100122.
  3. Nakashima, Kenichiro (2003). "Lophine derivatives as versatile analytical tools". Biomedical Chromatography. 17 (2–3): 83–95. doi:10.1002/bmc.226. PMID   12717796.
  4. Shah, Syed Niaz Ali; Lin, Jin-Ming (2017). "Recent advances in chemiluminescence based on carbonaceous dots". Advances in Colloid and Interface Science. 241: 24–36. doi:10.1016/j.cis.2017.01.003. PMID   28139217.
  5. "Luminol chemistry laboratory demonstration" . Retrieved 2006-03-29.
  6. "Investigating luminol" (PDF). Salters Advanced Chemistry. Archived from the original (PDF) on September 20, 2004. Retrieved 2006-03-29.
  7. Rauhut, Michael M. (1985), Chemiluminescence. In Grayson, Martin (Ed) (1985). Kirk-Othmer Concise Encyclopedia of Chemical Technology (3rd ed), pp 247 John Wiley and Sons. ISBN   0-471-51700-3
  8. Air Zoom | Glowing with Pride Archived 2014-06-12 at the Wayback Machine . Fannation.com. Retrieved on 2011-11-22.
  9. Atkins P. and de Paula J. Physical Chemistry (8th ed., W.H.Freeman 2006) p.886 ISBN   0-7167-8759-8
  10. Steinfeld J.I., Francisco J.S. and Hase W.L. Chemical Kinetics and Dynamics (2nd ed., Prentice-Hall 1998) p.263 ISBN   0-13-737123-3
  11. Atkins P. and de Paula J. p.889-890
  12. Enhanced CL review. Biocompare.com (2007-06-04). Retrieved on 2011-11-22.
  13. High Intensity HRP-Chemiluminescence ELISA Substrate Archived 2016-04-08 at the Wayback Machine . Haemoscan.com (2016-02-11). Retrieved on 2016-03-29.
  14. "Ecophysics CLD790SR2 NO/NO2 analyser" (PDF). Archived from the original (PDF) on 2016-03-04. Retrieved 2015-04-30.
  15. Stella, P., Kortner, M., Ammann, C., Foken, T., Meixner, F. X., and Trebs, I.: Measurements of nitrogen oxides and ozone fluxes by eddy covariance at a meadow: evidence for an internal leaf resistance to NO2, Biogeosciences, 10, 5997-6017, doi : 10.5194/bg-10-5997-2013, 2013.
  16. Tsokankunku, Anywhere: Fluxes of the NO-O3-NO2 triad above a spruce forest canopy in south-eastern Germany. Bayreuth, 2014 . - XII, 184 P. ( Doctoral thesis, 2014, University of Bayreuth, Faculty of Biology, Chemistry and Earth Sciences)
  17. Kinn, John J "Chemiluminescent kite" U.S. patent 4,715,564 issued 12/29/1987
  18. Kuntzleman, Thomas Scott; Rohrer, Kristen; Schultz, Emeric (2012-06-12). "The Chemistry of Lightsticks: Demonstrations To Illustrate Chemical Processes". Journal of Chemical Education. 89 (7): 910–916. Bibcode:2012JChEd..89..910K. doi:10.1021/ed200328d. ISSN   0021-9584.
  19. Chemiluminescence as a Combustion Diagnostic Archived 2011-03-02 at the Wayback Machine Venkata Nori and Jerry Seitzman - AIAA - 2008
  20. "Sustainable light achieved in living plants". phys.org. Retrieved 18 May 2020.
  21. "Scientists use mushroom DNA to produce permanently-glowing plants". New Atlas. 28 April 2020. Retrieved 18 May 2020.
  22. "Scientists create glowing plants using mushroom genes". the Guardian. 27 April 2020. Retrieved 18 May 2020.
  23. Mitiouchkina, Tatiana; Mishin, Alexander S.; Somermeyer, Louisa Gonzalez; Markina, Nadezhda M.; Chepurnyh, Tatiana V.; Guglya, Elena B.; Karataeva, Tatiana A.; Palkina, Kseniia A.; Shakhova, Ekaterina S.; Fakhranurova, Liliia I.; Chekova, Sofia V.; Tsarkova, Aleksandra S.; Golubev, Yaroslav V.; Negrebetsky, Vadim V.; Dolgushin, Sergey A.; Shalaev, Pavel V.; Shlykov, Dmitry; Melnik, Olesya A.; Shipunova, Victoria O.; Deyev, Sergey M.; Bubyrev, Andrey I.; Pushin, Alexander S.; Choob, Vladimir V.; Dolgov, Sergey V.; Kondrashov, Fyodor A.; Yampolsky, Ilia V.; Sarkisyan, Karen S. (27 April 2020). "Plants with genetically encoded autoluminescence". Nature Biotechnology. 38 (8): 944–946. doi:10.1038/s41587-020-0500-9. ISSN   1546-1696. PMC   7610436 . PMID   32341562. S2CID   216559981.