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Uv visible spectroscopy

UV-Vis Spectroscopy

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UV / VISIBLE SPECTROSCOPY
Mr. Santosh M. Damkondwar
May 3, 2018
Spectroscopy
• It is the branch of science that deals with the
study of interaction of matter with light.
OR
• It is the branch of science that deals with the
study of interaction of electromagnetic
radiation with matter.
Electromagnetic
Radiation
Electromagnetic Radiation
• Electromagnetic radiation consist of discrete
packages of energy which are called as
photons.
• A photon consists of an oscillating electric field
(E) & an oscillating magnetic field (M) which
are perpendicular to each other.
Uv visible spectroscopy
Electromagnetic Radiation
• Frequency (ν):
– It is defined as the number of times electrical field
radiation oscillates in one second.
– The unit for frequency is Hertz (Hz).
1 Hz = 1 cycle per second
• Wavelength (λ):
– It is the distance between two nearest parts of the
wave in the same phase i.e. distance between two
nearest crest or troughs.

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Uv visible spectroscopy

  • 1. UV / VISIBLE SPECTROSCOPY Mr. Santosh M. Damkondwar May 3, 2018
  • 2. Spectroscopy • It is the branch of science that deals with the study of interaction of matter with light. OR • It is the branch of science that deals with the study of interaction of electromagnetic radiation with matter.
  • 4. Electromagnetic Radiation • Electromagnetic radiation consist of discrete packages of energy which are called as photons. • A photon consists of an oscillating electric field (E) & an oscillating magnetic field (M) which are perpendicular to each other.
  • 6. Electromagnetic Radiation • Frequency (ν): – It is defined as the number of times electrical field radiation oscillates in one second. – The unit for frequency is Hertz (Hz). 1 Hz = 1 cycle per second • Wavelength (λ): – It is the distance between two nearest parts of the wave in the same phase i.e. distance between two nearest crest or troughs.
  • 7. Electromagnetic Radiation • The relationship between wavelength & frequency can be written as: c = ν λ • As photon is subjected to energy, so E = h ν = h c / λ
  • 9. Electromagnetic Radiation Violet 400 - 420 nm Yellow 570 - 585 nm Indigo 420 - 440 nm Orange 585 - 620 nm Blue 440 - 490 nm Red 620 - 780 nm Green 490 - 570 nm
  • 11. Principles of Spectroscopy • The principle is based on the measurement of spectrum of a sample containing atoms / molecules. • Spectrum is a graph of intensity of absorbed or emitted radiation by sample verses frequency (ν) or wavelength (λ). • Spectrometer is an instrument design to measure the spectrum of a compound.
  • 12. Principles of Spectroscopy 1. Absorption Spectroscopy: • An analytical technique which concerns with the measurement of absorption of electromagnetic radiation. • e.g. UV (185 - 400 nm) / Visible (400 - 800 nm) Spectroscopy, IR Spectroscopy (0.76 - 15 μm)
  • 13. Principles of Spectroscopy 2. Emission Spectroscopy: • An analytical technique in which emission (of a particle or radiation) is dispersed according to some property of the emission & the amount of dispersion is measured. • e.g. Mass Spectroscopy
  • 15. Interaction of EMR with matter 1. Electronic Energy Levels: • At room temperature the molecules are in the lowest energy levels E0. • When the molecules absorb UV-visible light from EMR, one of the outermost bond / lone pair electron is promoted to higher energy state such as E1, E2, …En, etc is called as electronic transition and the difference is as: ∆E = h ν = En - E0 where (n = 1, 2, 3, … etc) ∆E = 35 to 71 kcal/mole
  • 16. Interaction of EMR with matter 2. Vibrational Energy Levels: • These are less energy level than electronic energy levels. • The spacing between energy levels are relatively small i.e. 0.01 to 10 kcal/mole. • e.g. when IR radiation is absorbed, molecules are excited from one vibrational level to another or it vibrates with higher amplitude.
  • 17. Interaction of EMR with matter 3. Rotational Energy Levels: • These energy levels are quantized & discrete. • The spacing between energy levels are even smaller than vibrational energy levels. ∆Erotational < ∆Evibrational < ∆Eelectronic
  • 19. Lambert’s Law • When a monochromatic radiation is passed through a solution, the decrease in the intensity of radiation with thickness of the solution is directly proportional to the intensity of the incident light. • Let I be the intensity of incident radiation. x be the thickness of the solution. Then
  • 20. Lambert’s Law I dx dI  So, KI dx dI  Integrate equation between limit I = Io at x = 0 and I = I at x=l, We get, Kl I I  0 ln
  • 23. Beer’s Law • When a monochromatic radiation is passed through a solution, the decrease in the intensity of radiation with thickness of the solution is directly proportional to the intensity of the incident light as well as concentration of the solution. • Let I be the intensity of incident radiation. x be the thickness of the solution. C be the concentration of the solution. Then
  • 24. Beer’s Law IC dx dI . So, ICK dx dI .' Integrate equation between limit I = Io at x = 0 and I = I at x=l, We get, lCK I I .'ln 0 
  • 25. Beer’s Law lCK I I ..log303.2 0  lC K I I . 303.2 log 0  Where, AbsorbanceA I I 0 log E K  303.2 lCEA .. Molar extinction coefficient Beer’s Law
  • 26. Beer’s Law lCEA .. 0I I T  OR A I I T  0 loglog From the equation it is seen that the absorbance which is also called as optical density (OD) of a solution in a container of fixed path length is directly proportional to the concentration of a solution.
  • 27. PRINCIPLES OF UV - VISIBLE SPECTROSCOPY
  • 28. Principle • The UV radiation region extends from 10 nm to 400 nm and the visible radiation region extends from 400 nm to 800 nm. Near UV Region: 200 nm to 400 nm Far UV Region: below 200 nm • Far UV spectroscopy is studied under vacuum condition. • The common solvent used for preparing sample to be analyzed is either ethyl alcohol or hexane.
  • 30. The possible electronic transitions can graphically shown as:
  • 31. • σ → σ* transition1 • π → π* transition2 • n → σ* transition3 • n → π* transition4 • σ → π* transition5 • π → σ* transition6 The possible electronic transitions are
  • 32. • σ electron from orbital is excited to corresponding anti-bonding orbital σ*. • The energy required is large for this transition. • e.g. Methane (CH4) has C-H bond only and can undergo σ → σ* transition and shows absorbance maxima at 125 nm. • σ → σ* transition1
  • 33. • π electron in a bonding orbital is excited to corresponding anti-bonding orbital π*. • Compounds containing multiple bonds like alkenes, alkynes, carbonyl, nitriles, aromatic compounds, etc undergo π → π* transitions. • e.g. Alkenes generally absorb in the region 170 to 205 nm. • π → π* transition2
  • 34. • Saturated compounds containing atoms with lone pair of electrons like O, N, S and halogens are capable of n → σ* transition. • These transitions usually requires less energy than σ → σ* transitions. • The number of organic functional groups with n → σ* peaks in UV region is small (150 – 250 nm). • n → σ* transition3
  • 35. • An electron from non-bonding orbital is promoted to anti-bonding π* orbital. • Compounds containing double bond involving hetero atoms (C=O, C≡N, N=O) undergo such transitions. • n → π* transitions require minimum energy and show absorption at longer wavelength around 300 nm. • n → π* transition4
  • 36. •These electronic transitions are forbidden transitions & are only theoretically possible. •Thus, n → π* & π → π* electronic transitions show absorption in region above 200 nm which is accessible to UV-visible spectrophotometer. •The UV spectrum is of only a few broad of absorption. • σ → π* transition5 • π → σ* transition 6&
  • 37. Terms used in UV / Visible Spectroscopy
  • 38. Chromophore The part of a molecule responsible for imparting color, are called as chromospheres. OR The functional groups containing multiple bonds capable of absorbing radiations above 200 nm due to n → π* & π → π* transitions. e.g. NO2, N=O, C=O, C=N, C≡N, C=C, C=S, etc
  • 39. Chromophore To interpretate UV – visible spectrum following points should be noted: 1. Non-conjugated alkenes show an intense absorption below 200 nm & are therefore inaccessible to UV spectrophotometer. 2. Non-conjugated carbonyl group compound give a weak absorption band in the 200 - 300 nm region.
  • 40. Chromophore e.g. Acetone which has λmax = 279 nm and that cyclohexane has λmax = 291 nm. When double bonds are conjugated in a compound λmax is shifted to longer wavelength. e.g. 1,5 - hexadiene has λmax = 178 nm 2,4 - hexadiene has λmax = 227 nm CH3 C CH3 O O CH2 CH2 CH3 CH3
  • 41. Chromophore 3. Conjugation of C=C and carbonyl group shifts the λmax of both groups to longer wavelength. e.g. Ethylene has λmax = 171 nm Acetone has λmax = 279 nm Crotonaldehyde has λmax = 290 nm CH3 C CH3 O CH2 CH2 C CH3 O CH2
  • 42. Auxochrome The functional groups attached to a chromophore which modifies the ability of the chromophore to absorb light , altering the wavelength or intensity of absorption. OR The functional group with non-bonding electrons that does not absorb radiation in near UV region but when attached to a chromophore alters the wavelength & intensity of absorption.
  • 43. Auxochrome e.g. Benzene λmax = 255 nm Phenol λmax = 270 nm Aniline λmax = 280 nm OH NH2
  • 46. • When absorption maxima (λmax) of a compound shifts to longer wavelength, it is known as bathochromic shift or red shift. • The effect is due to presence of an auxochrome or by the change of solvent. • e.g. An auxochrome group like –OH, -OCH3 causes absorption of compound at longer wavelength. • Bathochromic Shift (Red Shift)1
  • 47. • In alkaline medium, p-nitrophenol shows red shift. Because negatively charged oxygen delocalizes more effectively than the unshared pair of electron. p-nitrophenol λmax = 255 nm λmax = 265 nm • Bathochromic Shift (Red Shift)1 OH N + O - O OH - Alkaline medium O - N + O - O
  • 48. • When absorption maxima (λmax) of a compound shifts to shorter wavelength, it is known as hypsochromic shift or blue shift. • The effect is due to presence of an group causes removal of conjugation or by the change of solvent. • Hypsochromic Shift (Blue Shift)2
  • 49. • Aniline shows blue shift in acidic medium, it loses conjugation. Aniline λmax = 280 nm λmax = 265 nm • Hypsochromic Shift (Blue Shift)2 NH2 H + Acidic medium NH3 + Cl -
  • 50. • When absorption intensity (ε) of a compound is increased, it is known as hyperchromic shift. • If auxochrome introduces to the compound, the intensity of absorption increases. Pyridine 2-methyl pyridine λmax = 257 nm λmax = 260 nm ε = 2750 ε = 3560 • Hyperchromic Effect3 N N CH3
  • 51. • When absorption intensity (ε) of a compound is decreased, it is known as hypochromic shift. Naphthalene 2-methyl naphthalene ε = 19000 ε = 10250 CH3 • Hypochromic Effect4
  • 52. Wavelength ( λ ) Absorbance(A) Shifts and Effects Hyperchromic shift Hypochromic shift Red shift Blue shift λmax
  • 53. APPLICATIONS OF UV / VISIBLE SPECTROSCOPY
  • 54. Applications • Qualitative & Quantitative Analysis: – It is used for characterizing aromatic compounds and conjugated olefins. – It can be used to find out molar concentration of the solute under study. • Detection of impurities: – It is one of the important method to detect impurities in organic solvents. • Detection of isomers are possible. • Determination of molecular weight using Beer’s law.
  • 56. Reference Books • Introduction to Spectroscopy – Donald A. Pavia • Elementary Organic Spectroscopy – Y. R. Sharma • Physical Chemistry – Puri, Sharma & Pathaniya