ABSTRACT Ultraviolet and visible spectroscopy (UV-vis) is a reliable and accurate analytical laboratory assessment procedure that allows

for the analysis of a substance. Specifically, ultraviolet and visible spectroscopy measures the absorption, transmission and emission of ultraviolet and visible light wavelengths by matter.In the chemistry laboratory, ultraviolet and visible spectroscopy (UV-vis spectroscopy) is used to study molecules and inorganic ions in solution. Although they are distinct regions of the electromagnetic spectrum, the ultraviolet and visible regions of the electromagnetic spectrum are linked in UV-vis spectroscopy because similarities between the two regions allow many of the same research techniques and tools to be used for both regions. Ultraviolet/visible spectroscopy is useful as an analytical technique for two reasons. First it can be used to identify some functional groups in molecules and secondly, it can be used for assaying. This second role determining the content and strength of a substance is extremely useful. Ultraviolet/visible spectroscopy is used extensively in chemical and biochemical laboratories, for a variety of tasks. It can be used to determine small quantities such as the trace metal content in alloys or the amount of a certain drug reaching various parts of the body.

INTRODUCTION

History The foundation of UV-visible spectroscopy was laid by English physicist and mathematician Sir Isaac Newton who, by diffracting white light through a prism, showed that it consisted of light of several colors ranging from violet to red. This is now termed the UV-visible electromagnetic spectrum. German physicist Maxwell Planck (1858-1947) first postulated that the energy of the electromagnetic spectrum was not continuous, but consisted of small, discrete packets which he called quanta, and whose energy, E, was directly proportional to the frequency of the light as expressed in the fundamental equation of spectroscopy, i.e. E = h where h is a proportionality constant called Planck's constant. The direct relationship between energy and frequency shows that light of lower frequency will carry fewer energetic photons than light of a higher frequency. Ultraviolet and visible spectrophotometric methods were pioneered by American chemist Arnold Beckman, a member of the National Inventors Hall of Fame. Beckman had an early love for chemistry and completed college chemistry courses

before he began his undergraduate studies at the University of Illinois. After completing his Ph.D. at the California Institute of Technology in 1928, Beckman went on to invent many measurement devices of scientific importance. In the 1940s, he invented a spectrophotometer that had the ability to measure the transmission of visible and ultraviolet and visible light through a target. UV-visible spectroscopy has been enhanced as an experimental technique by the introduction of lasers. This field was developed in the 1950s with the initial work occurring in the microwave region, but it quickly extended to the UV-visible regions. A commonly used laser today is the ruby laser, in which the lasing medium is a rod of sapphire (AlO) with some ions replaced by Cr(III) ions, giving the crystal its bright red color.

THEORY For most spectra the solution obeys Beers Law. This states that the light absorbed is proportional to the number of absorbing molecules ie to the concentration of absorbing molecules. This is only true for dilute solutions. A second law Lamberts law tells us that the fraction of radiation absorbed is independent of the intensity of the radiation. Combining these two laws gives the BeerLambert law: Absorbance = log10I0/I = lc IO = the intensity of the incident radiation I = the intensity of the transmitted radiation = the molar absorption coefficient l = the path length of the absorbing solution (cm) c = the concentration of the absorbing species in mol dm-3 Two useful pieces of information are the molar absorption coefficient, and max which is the wavelength at which maximum absorption occurs. These two pieces of information are sometimes enough to identify a substance. However, if and max are known for a compound the concentration of the solution can be calculated. This is the most common application.

How It Works

Samples are used in solution and are placed in a small silica cell. Two lamps are used. A hydrogen or deuterium lamp for the ultraviolet region and a tungsten/halogen lamp for the visible region. In this way radiation across the whole range is scanned by the spectrometer . A reference cell containing only solvent is used. Light is passed simultaneously through the sample cell and reference cell. The spectrometer compares the light passing through the sample with that passing through the reference cell. The transmitted radiation is detected and the spectrometer records the absorption spectrum by scanning the wavelength of the light passing through the cells. A schematic diagram is shown in Fig. 1. Single beam instruments are now available as well as double beam instruments. Radiation across the whole range is monitored simultaneously. The principles are the same as for double beam instruments, but data on the reference are taken first, followed by the sample. In this way single beam instruments can record the spectrum very quickly.

Figure 1: Schematic diagram

Worked Example The UV-vis spectrum of tetraphenyclopentadienone is given below and should be similar to the one you obtained from lab. The "n" electrons (or the nonbonding electrons) are the ones located on the oxygen of the carbonyl group of tetraphenyclopentadienone. Thus, the n to pi* transition corresponds to the excitation of an electron from one of the unshared pair to the pi* orbital. Figure 2 : UV-Vis analysis of Tetraphenylcyclopentadienone

REFERENCES Beckman, A.O.; Gallaway, W. S.;Kaye, W.; Ulrich, W.F. History of Spectrophotometry at Beckman Instruments,Inc. Anal.Chem. 1977,49(3),280 A-300 A Cary, H.H.; Beckhman, A.O.,Jr. A Quartz Photoelectric Spectrophotometer. J.Opt. Soc.Am.1941,31,686-689 NIST Chemistry Web Book. Htpp:webbook.nist.gov/chemistry/First accessed 11/100. http://www.bookrags.com/Ultraviolet-visible_spectroscopy