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Introduction UV spectroscopy involves the measurement of absorption of light in the visible and ultraviolet regions (visible region 400- 800 nm ; uv region 200-400nm) by the substance under investigation. Since the absorption of light involves the transition from one electronic energy level to another within a molecule, UV spectroscopy is also known as electronic spectroscopy. This technique is complementary to fluorescence spectroscopy, in that fluorescence deals with transitions from the excited state to the ground state, while absorption measures transitions from the ground state to the excited state. Principle of absorption spectroscopy Molecules containing π-electrons or non-bonding electrons (n- electrons) can absorb the energy in the form of ultraviolet or visible light to excite these electrons to higher anti-bonding molecular orbitals. The more easily excited the electrons (i.e. lower energy gap between the HOMO and the LUMO), the longer the wavelength of light it can absorb. A very important condition for a molecule to absorb electromagnetic radiation is that the energy of photon of radiation must be equal to the energy difference between two vibrational or rotational or electronic energy states of the molecule. A record of the amount of radiation absorbed or transmitted by a given sample as function of wavelength of radiation is called absorption spectrum. Beer’s Lambert law When a beam of monochromatic light is passed through a substance dispersed in a non-absorbing solvent, absorption of light is directly proportional to molar concentration of the absorbing substance as well as path length of the sample substance. Using the Beer-Lambert law: where A is the measured absorbance, in Absorbance Units (AU), is the intensity of the incident light at a given wavelength, is the transmitted intensity, L the path length through the sample, and c the concentration of the absorbing species. For each species and wavelength, ε is a constant known as the molar absorptivity or extinction coefficient. Electronic excitations sigma to sigma* transition : very high energy required consequently occur at short wavelength. Eg: methane. n to sigma* transition: occur at long wavelength than sigma to sigma*. Eg: methyl chloride . n to pi*transition: require small amount of energy and take place within the range of ordinary uv spectrophotometer. Eg : carbonyl group of saturated aldehydes and ketones. pi to pi* transitions : relatively high energy requirement than n to pi * transitions and absorption generally takes place outside the ordinary uv region. Eg : aldehydes and ketones. Effect of conjugation conjugation of double bonds lowers the energy required for electronic transition, molecules containing conjugated systems absorb radiations of longer wavelength than in case of non conjugated systems. Eg: 1,3-butadiene shows max at 217 nm in contrast to ethylene which shows at 175nm. energy gap between HOMO and LUMO decreases. as the gap decreases position of energy moves to longer wavelength which falls within the ordinary uv region.
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