Chemiluminescence in Analytical Chemistry

Contributors

Hassan Y. Aboul-Enein Pharmaceutical Analysis Laboratory, Department of Biological and Medical Research, King Faisal Specialist Hospital and Research Centre, Riyadh, Saudi Arabia

Willy R. G. Baeyens Department of Pharmaceutical Analysis, Ghent University, Ghent, Belgium

Mario Baraldini Institute of Chemical Sciences, University of Bologna, Bologna, Italy

John W. Birks Department of Chemistry and Biochemistry and Cooperative Institute for Research in Environmental Sciences, University of Colorado, Boulder, Colorado

Luca Bolelli Institute of Chemical Sciences, University of Bologna, Bologna,

Italy

Richard Bos School of Biological and Chemical Sciences, Deakin University, Geelong, Victoria, Australia

James E. Boulter Department of Chemistry and Biochemistry, University of Colorado, Boulder, Colorado

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xii	Contributors

Antony C. Calokerinos Laboratory of Analytical Chemistry, Department of Chemistry, University of Athens, Athens, Greece

Elida Nora Ferri Institute of Chemical Sciences, University of Bologna, Bologna, Italy

Fabiana Fini Institute of Chemical Sciences, University of Bologna, Bologna,

Italy

Ana M. Garcı´a-Campan˜a Department of Analytical Chemistry, University of Granada, Granada, Spain

Stefano Girotti Institute of Chemical Sciences, University of Bologna, Bologna, Italy

Massimo Guardigli Department of Pharmaceutical Sciences, University of Bologna, Bologna, Italy

Norberto A. Guzman Department of Bioanalytical Drug Metabolism, The R.W. Johnson Pharmaceutical Research Institute, Raritan, New Jersey

Knut Irgum Department of Analytical Chemistry, Umea˚ University, Umea˚,

Sweden

Marjorie Jacquemijns National Institute of Public Health and the Environment, Bilthoven, The Netherlands

Tobias Jonsson Department of Analytical Chemistry, Umea˚ University, Umea˚,

Sweden

Masaaki Kai Department of Medicinal Chemistry, School of Pharmaceutical Sciences, Nagasaki University, Nagasaki, Japan

Andrew W. Knight Department of Instrumentation and Analytical Science, University of Manchester Institute of Science and Technology, Manchester, England

Naotaka Kuroda Department of Analytical Chemistry, School of Pharmaceutical Sciences, Nagasaki University, Nagasaki, Japan

Dan A. Lerner Department of Physical Chemistry, Ecole Nationale Supe´rieure de Chimie, Montpellier, France

Contributors

xiii

Gudrun Lewin Research Institute for Antioxidant Therapy, Berlin, Germany

Mara Mirasoli Department of Pharmaceutical Sciences, University of Bologna, Bologna, Italy

Monica Musiani Division of Microbiology, Department of Clinical and Experimental Medicine, University of Bologna, Bologna, Italy

Kenichiro Nakashima Department of Analytical Research for Pharmacoinformatics, Graduate School of Pharmaceutical Sciences, Nagasaki University, Nagasaki, Japan

Kazuko Ohta School of Pharmaceutical Sciences, Nagasaki University, Nagasaki, Japan

Leonidas P. Palilis Laboratory of Analytical Chemistry, Department of Chemistry, University of Athens, Athens, Greece

Patrizia Pasini Department of Pharmaceutical Sciences, University of Bologna, Bologna, Italy

Dolores Pe´rez-Bendito Department of Analytical Chemistry, University of Co´rdoba, Co´rdoba, Spain

Einar Ponte´n Department of Analytical Chemistry, Umea˚ University, Umea˚,

Sweden

Igor Popov Research Institute for Antioxidant Therapy, Berlin, Germany

Yener Rakiciog˘lu Department of Chemistry, Istanbul Technical University,

Istanbul, Turkey

Aldo Roda Division of Analytical Chemistry, Department of Pharmaceutical Sciences, University of Bologna, Bologna, Italy

Manuel Roma´n-Ceba Department of Analytical Chemistry, University of Granada, Granada, Spain

Carmela Russo Department of Pharmaceutical Sciences, University of Bologna, Bologna, Italy

xiv	Contributors

Jose´ Juan Santana Rodrı´guez Department of Chemistry, University of Las Palmas de G.C., Las Palmas de G.C., Spain

Joanna M. Schulman Department of Botany, University of Florida, Gainesville, Florida

Stephen G. Schulman Department of Medicinal Chemistry, College of Pharmacy, University of Florida, Gainesville, Florida

Gloria Sermasi Institute of Chemical Sciences, University of Bologna, Bologna, Italy

Manuel Silva Department of Analytical Chemistry, University of Co´rdoba, Co´rdoba, Spain

Raluca-Ioana Stefan Department of Chemistry, University of Pretoria, Pretoria, South Africa

Malin Stigbrand Department of Analytical Chemistry, Umea˚ University,

Umea˚, Sweden

Jacobus F. van Staden Department of Chemistry, University of Pretoria, Pretoria, South Africa

Xinrong Zhang Department of Chemistry, Tsinghua University, Beijing, P. R.

China

Gijsbert Zomer National Institute of Public Health and the Environment, Bilthoven, The Netherlands

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Historical Evolution of Chemiluminescence

Ana M. Garcı´a-Campan˜a and Manuel Roma´n-Ceba

University of Granada, Granada, Spain

Willy R. G. Baeyens

Ghent University, Ghent, Belgium

1.	INTRODUCTION TO THE DISCOVERY AND THE
	DEVELOPMENT OF LUMINESCENCE		2
2.	THE DISCOVERY OF BIOLUMINESCENCE AND CHEMI-
	LUMINESCENCE PHENOMENA		11
3.	STUDY OF THE CHEMILUMINESCENT SYSTEMS		12
4.	THE FIRST ANALYTICAL USES OF BIOLUMINESCENCE
	AND CHEMILUMINESCENCE		22
	4.1	Chemiluminescence in the Gas Phase	22
	4.2	Chemiluminescent Systems as Indicators in Titrations	24
	4.3	Application to Firefly Luciferase Reaction in the Analysis of
		ATP	26
	4.4	The Application of Chemiluminescence in Immunoassay	27
	4.5	Chemiluminescence in Flow Analysis and Immobilization
		Techniques	28
	4.6	First Uses of Chemiluminescence as a Detection Technique in
		HPLC	29
	4.7	Application of Chemiluminescence in DNA Analysis	30
	4.8	Chemiluminescence Sensors	31
	4.9	Chemiluminescence Produced by Direct Oxidation	31
5.	CONCLUSIONS		32

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1.INTRODUCTION TO THE DISCOVERY AND THE DEVELOPMENT OF LUMINESCENCE

It has been known for centuries that many compounds emit visible radiation when they are exposed to sunlight. Luminescence phenomena, such as the aurora borealis, phosphorescence of the sea, luminous animals and insects, phosphorescent wood, etc., have fascinated man since antiquity, being reflected in the early scientific literature. Aristotle (384–322 B.C.) appears to be one of the first philosophers to recognize ‘‘cold light’’ in dead fish, fungi, and the luminous secretion of the cuttlefish [1].

These luminescence phenomena have been known since ancient times; according to the legend, about 1000 B.C., a Chinese emperor possessed a magic paint on which the image of an ox appeared at sunset. The chemical composition of the paint used was not known. This is the first known case of a man-made substance capable of storing daylight for later recovery [2].

Also, early written references citing luminescence phenomena appeared in the Chinese literature of around 1500–1000 B.C., describing glowworms and fireflies [3]. In fact, all of these first observations were related mainly to living organisms that emit light such as the fireflies, luminous bacteria and protozoa, the sea pansy, the marine fireworm, unicellular organisms such as the dinoflagellates, etc. Harvey describes in his book an interesting chapter on the many early approaches to explain luminescence, as a matter of fact an established modern scientific approach on the subject, from the seventeenth century [3].

Francis Bacon reports in 1605 the different kinds of luminescence in relation to its origins and writes: ‘‘sugar shineth only while it is scraping; and salt water while it is in dashing; glowworms have their shining while they live, or a little after; only scales of fish putrefied seem to be of the same nature with shining wood: and it is true, that all putrefaction has with it an inward motion, as well as fire or light’’ [3].

The first example of luminescence emission from solids, of which written documents exist, date from the Italian Renaissance, originating from the accidental discovery around the year 1600 (1602 or 1603) by a Bolonian shoemaker and alchemist, called Vincencio Casciarolo or Casciarolus. He melted heavy bricks, close to his house, hoping to extract precious metals from them.

These bricks, after calcination with carbon and exposure to daylight, emitted a reddish glittering in the dark. These ‘‘Bolonian stones,’’ also named ‘‘moonstones,’’ particularly those from the Monte Paterno, remain among the most famous ones and were the subject of scientific interest during the next two centuries; they were termed ‘‘phosphor’’ (Greek: ‘‘light bearer’’). They are considered the first inorganic artificial ‘‘phosphors’’ [2–4]. The first natural phosphor was diamant, whose luminescence was cited by Cellini in 1568 [5].

Historical Evolution of Chemiluminescence

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The discovery of the ‘‘Bolonian stones’’ attracted the interest of Galileo (1564–1642) and colleagues from that period who stated that a ‘‘phosphor’’ does not emit luminescence before having been exposed to natural light, in the way that light enters the stone, like a sponge taking up water, then producing light emission, so that the ‘‘phosphor’’ behavior implicated some time during which light remained for a while in the substance material [6]. In 1652 the Italian mathematician Zucchi described that the Bolonian stone emitted more intensely once exposed to brilliant light and that the color of the emitted light did not change when the stone was exposed to white light, green, yellow, or red light. He concluded that the light was not simply absorbed and emitted in an unchanged form, like a sponge, but that on the contrary, during the excitation process reactions occurred with some substance (‘‘spiritus’’) present in the brick, and that when illumination ceased, the light produced by the substance gradually diminished [3].

Originally, the Bolonian stone, as mentioned, was considered a ‘‘light sponge,’’ but clearly this term was poetic rather than exact. In this poetic way, as a matter of fact, the poet Goethe described the stone. When light of different colors impinged upon this stone, some red light was produced when exciting with blue light, red light as such being unable to do so. The light emitted by the stone in the dark did not appear to be similar to the illuminating light applied, which made clear that the Bolonian stone in fact did not act as a sponge that post factum emitted the absorbed light [7].

The delayed light emission as observed from the Bolonian stone is now classified as phosphorescence. We know now that these stones contain barium sulfate with traces of bismuth and manganese, and that the corresponding reducing process concerns the transformation of sulfate into sulfur. It is now well known that alkaline earth metal sulfates emit phosphorescence that strongly increases when traces of heavy metals are present. The so-called inorganic multicomponent compounds ‘‘phosphor’’ and ‘‘crystallophosphor’’ are in fact polycrystalline substances containing traces of some ionic activators of luminescence.

The term ‘‘phosphor’’ obviously is employed as well for the chemical element discovered by a Hamburg alchemist, Hennig Brand, in 1669, recognized to be the first scientist discovering a chemical element. In fact Brand, searching for the philosopher’s stone, distilled a mixture of sand and evaporated urine and obtained a product that was capable of shining in the dark. They called it ‘‘Brand’s phosphor’’ to distinguish it from other luminescent materials also termed ‘‘phosphor’’ [8]. Brand called the product obtained ‘‘miraculous light’’ [3]. The element P, in its white or yellow allotropic variety, emits light in the dark, but this is not a photoluminescent (phosphorescent) phenomenon but a chemiluminescence produced by the reaction of this element with oxygen, in a humid environment [9].

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The first example of protein luminescence was made by Beccari in 1746, who detected a visible, blue phosphorescence proceeding from frozen hands when entering a dark room after exposure to sunlight [10].

During the seventeenth and eighteenth centuries, numerous ‘‘phosphors’’ were discovered, but little progress in their characterization occurred. An attempt to classify luminescent phenomena, indicated by the general term ‘‘phosphorescence,’’ appears at the end of the eighteenth century. The Encyclopedia of Diderot and Alembert, in its Geneva edition of 1778–1779, mentions six classes of ‘‘phosphors’’ differentiating the slow oxidation of the metalloid from: physiological phenomena (fireflies, glowworms, mosquitos from the Venice lagoon, flies from the Antilles, sting of irritated vipers); electric phenomena (diamond, strongly rubbed tissues and clothes, Hauxbee globe); mechanical effects (friction of sugar or cadmia, metals trapped in steel or iron); physical phenomena (Bolonian stone and spar exposed to sunlight); biological phenomena (will-o’-the wisp) [4].

The first observation of fluorescence in solution occurred in 1565 by the Spanish physician and botanist Nicola´s Monardes, who noticed a blue tint in the water contained in a recipient fabricated with a specimen of wood called ‘‘lignum nephriticum’’ [3, 11]. It was known in 1570 that the blue coloration that is produced by white light from the aqueous extract of the ‘‘lignum nephriticum’’ or ‘‘peregrinum’’ disappeared in acid medium. In 1615 a similar behavior was observed from the rind of Aesculos hippocastanum in aqueous medium [4]. When placed in water, the rind of chestnut produces a colorless liquid with bluish reflections; today it is known that this originates from aesculin fluorescence [12].

The luminescent properties of the aqueous extracts of some wood specimens were of interest to many scientists of the seventeenth and the beginning of the eighteenth century. Athanasius Kircher, who investigated aqueous extracts of wood, postulated in 1646 that the observed color depended on the intensity of the ambient light [3]. Robert Boyle (1627–1691), in 1664, Isaac Newton (1662–1727), and Robert Hooke, in 1678, disagreed with Kircher, who indicated that the color of the wood extract depended on the angle of observation, being yellow for the transmitted light and blue for the reflected light [6].

In the year 1704, in an optical treatise Newton stated that the tincture of lignum nephriticum showed a variable color depending on the position of the sun and of the incident light: yellow by transparency and blue from a lateral view [4]. Although Kircher is generally recognized as the discoverer of the fluorescence phenomenon in solution, it was Boyle who was the first to describe some of the most important characteristics of fluorescence from organic solutions. Boyle, after carrying out various extractions of wood, obtained a fluorescent solution and thought this to be an ‘‘essential salt’’ present in the wood, responsible for the observed luminescence. All cited scientists describe the production of a certain ‘‘reflection’’ phenomenon, without clearly providing any differentiation between the terms ‘‘emission’’ and ‘‘reflection’’ [3]. Hooke, in 1665, mentions that be-

Historical Evolution of Chemiluminescence

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cause of internal vibrations, some matrices emit light [3]. In 1718 Newton produces a report agreeing with Hooke’s hypothesis, stating that the incandescence of luminous bodies—hot or cold—originates from vibrational movements of their particles [13].

Although numerous materials and fluorescing solutions were described in the seventeenth and eighteenth centuries, and in spite of the fact that since around 1860 mineralogists started the use of fluorescence for detection of mineral deposits, little progress was observed concerning the explanation of the phenomenon, and it was only around the mid-nineteenth century that important achievements were made in the study and understanding of luminescence phenomena.

In 1833 David Brewster (1781–1868) describes the red fluorescence from an alcoholic extract from green leaves (i.e., chlorophyll) and the fluorescence from fluorspar crystals, but he considered the effect to be caused by ‘‘dispersed’’ light, rather than by emitted light. John Herschel (1792–1871), in 1845, uses a prism to obtain crude spectral analysis of the fluorescence from quinine solutions. Apparently he did not realize that the emitted light had a longer wavelength. He observed that the solution emitted a noticeable luminescent radiation when exposed to sunlight. The solution of quinine sulfate was colorless when observed by transparency and bluish white when examined from a certain angle. He postulated that the blue light was produced at the surface of the liquid and called this phenomenon ‘‘dispersion epipolique.’’ He had already suggested in 1825 that ‘‘dispersed’’ light might be employed for the detection of small quantities of some compounds. His compatriot Brewster, who studied the dissolution of quinine and of aesculine, mentioned that the ‘‘dispersion’’ occurred much more internally than superficially. Hence, the concepts of absorption and of emission as well as the phenomenon of fluorescence had not been established yet [5, 14]. It is important to note that the ideas of light absorption and emission were suggested much earlier for the ‘‘phosphors’’ (seventeenth century) than for fluorescent materials (nineteenth century), probably because understanding of the phosphorescence emission phenomenon occurred only later [5].

Sir George Gabriel Stokes (1820–1903), physicist and professor of mathematics at Cambridge, bears the merit of having established the theoretical principles of fluorescence, in an important publication in the journal Philosophical Transactions that appeared in the year 1852. By means of a setup of prisms he obtained a solar spectrum that he utilized to illuminate a tube containing a solution of quinine sulfate in the way that the red, yellow, green, etc. light passed the solution. When coming close to the violet or further spectral zones, a blue shining was progressively produced by the solution. It is extraordinary, described Stokes, to see how the tube is illuminated instantaneously by the ‘‘invisible rays.’’ These rays are what is today called ultraviolet radiation. He stated that the blue light in fact was made by the material starting from other radiations that were absorbed by the liquid, in the way that light production was more or less important