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  • Print publication year: 2007
  • Online publication date: November 2012

Chapter E1 - Visible and IR absorption spectroscopy

from Part E - Optical spectroscopy


Brief historical review and biological applications


In OpticksIsaac Newton dealt with the formation of a spectrum by a prism, and the composition of white light and its dispersion. The Latin word, spectrum, means an appearance; a spectrum is obtained when radiation is broken up into its colour or wavelength distribution.


The astronomer, William Herschel, discovered infrared (IR) radiation.


The physicist Johann Wilhelm Ritter discovered ultraviolet (UV) radia-tion.


Joseph von Frauenhofer showed that the Sun's spectrum contained dark lines (later named Frauenhofer lines), indicating that light of the corresponding colour was missing because of absorption.


August Beer stated the empirical law, which was named after him, that there is an exponential dependence between the transmision of light through a substance, the concentration of the substance and the path length of the beam through it. The law is also known as the Beer–Lambert law or the Beer–Lambert–Bouguer law, in recognition of the work of Pierre Bouguer (1729) and Johann Heinrich Lambert (1760). Gustav Kirchhoff's discovery that each pure substance has a characteristic spectrum provided the basis for analytical spectroscopy. Gustav Kirchhoff and Robert Bunsen identified the chemical elements in the Sun by analysing its spectrum. Johann Jacob Balmer identified a numerical series in the spectrum of hydrogen. Joseph John Thomson discovered the electron. Max Planck introduced the concept of quanta in the treatment of heat radiation and laid the foundation of quantum theory. He was awarded the Nobel prize in 1918.

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Suggestions for further reading
Historical review
Seibert, F. B. (1995). Infrared spectroscopy applied to biochemical and biological problems. Meth. Enzymol., 246, 501–526.
Chalmers, J. M., and Griffiths, P. R. (eds.) (2002). Handbook of Vibrational Spectroscopy, 5 volumes. Chichester: John Wiley and Sons Ltd.
Gauglitz, G., and Vo-Dinh, T. (eds.) (2003). Handbook of Spectroscopy, 2 volumes. Weinheim: Wiley-VCH Verlag GmbH KgaA.
Jakson, M., Sowa, M. G., and Mantsch, H. H. (1997). Infrared spectroscopy: a new frontier in medicine. Biophys. Chem., 68, 109–125.
Tinoco, I. Jr. (1995). Optical spectroscopy: General principles and overview. Meth. Enzymol., 246, 13–18.
Brief theoretical outline
Taillandier, E., Firon, M., and Liquier, J. (1991). In Spectroscopy of Biological Macromolecules, eds. Hester, R. E. and Girling, R. B.. Cambridge: Royal Society of Chemistry.
Mathies, R. (1995). Biomolecular vibrational spectroscopy. Meth. Enzymol., 246, 377–389.
Sauer, K. (1995). Why spectroscopy? Which spectroscopy? Meth. Enzymol., 246, 1–10.
Tinoco, I. Jr, Sauer, K., and Wang, J. C. (1998). Molecular structure and interactions: physical chemistry. In Principles and Applications in Biological Science. Ch. 10. Molecular structures and interactions: Spectroscopy. Upper Saddle River, NJ: Prentice Hall.
The UV–visible spectral range
Miura, T., and Thomas, G. J. Jr, (1995). Optical and vibrational spectroscopic methods. In Introduction to Biophysical Methods for Protein and Nucleic Acid Research, eds. Glasel, J. A. and Deutcher, M. P.. London: Academic Press.
Miyazawa, T., and Blout, E. R. (1961). The infrared spectra of polypeptides in various conformations: Amide I and II bands. JACS, 83, 712–719.
Heberle, J., and Gensch, Th. (2001). When FT-IR spectroscopy meets X-ray crystallography. Nature Struct. Biol., 8, 195–197.
Surevicz, W. K., Mantsch, H. H., and Chapman, D. (1993). Determination of protein secondary structure by Fourier transform infrared spectroscopy: a critical assessment. Biochemistry, 32, 389–394.
Susi, H., and Byler, D. M. (1986). Resolution-enhanced Fourier transform infrared spectroscopy of enzymes. Meth. Enzymol. 130, 290–311.
Braiman, M. S., and Rotschild, K. J. (1988). Fourier transform infrared techniques for probing membrane protein structure. Ann. Rev. Biophys. Chem., 17, 541–570.
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Martin, J-L., and Vos, M. H. (1992). Femtosecond biology. Annu. Rev. Biophys. Biomol. Struct., 21, 199–222.
Gevert, K. (1993). Molecular reaction mechanisms of proteins as monitored by time-resolved FTIR spectroscopy. Curr. Opin. Struct. Biol., 3, 769–773.
Taillandier, E., and Liquier, J. (1992). Infrared spectroscopy of DNA. Meth. Enzymol., 211, 307–352.