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Infrared spectrophotometry
-Very important in modern organic pharmaceutical chemistry. -Deals with infrared region of electromagnetic spectrum. -A type of absorption spectroscopy. -Routine tool for determining functional groups, identifying compounds & analysing mixtures. -Hence, When infrared light is passed through a sample of an organic compound, some of the frequencies are absorbed, while other frequencies are transmitted through the sample without being absorbed. -When absorbance or transmittance are plotted against frequency, the result is an infrared spectrum. -The IR region of the electromagnetic spectrum may be divided into three main sections: 1)Near IR(overtone region) 0.8-2.5m(12,500 4,000cm-1) 2)Middle-IR(vibration-rotation region) 2.5 - 50 m(4,000-200cm-1) 3)Far-IR (rotation region) 50-1000m(200-10cm-1) Units and relation: ___ 1____= Wavenumber= Frequency(Hz) Wavelength(m) (cm-1) Velocity of light(cm/s) 1/ = = /c Wavenumber (cm-1 )= 104 / m The units in common use are: 1Angstrom unit = 1A = 10-10metre = 10-8cm 1nanometre = 1nm = 10A = 10-7cm 1micrometre = 1m = 104A= 10-4cm Velocity of light = c = 3 X 108ms-1 -The fundamental region is between 2.5-25m (4000 400cm-1). -It provides the greatest information for the elucidation of molecular structure. -Most IR spectrophotometers are limited to measurements in this region.

Working principle: Absorption of IR takes place: a)When the energy of the radiation exactly matches the difference in energy level E between the vibrational quantum number. b)Dipole Moment Changes during Vibrations & Rotations. -The relative positions of atoms in a molecules fluctuate continuously. -Modes of movement are rotations & vibrations. (Origin of IR spectra) -IR radiation is not enough to bring about electronic transitions as in ultraviolet & visible radiation. -IR corresponds to molecular species with small energy differences between various vibrational & rotational states.

-Each vibrational level comprises many rotational levels. -Matching of radio frequency to natural(resonant frequency) is followed by absorption of radiation. -A molecule must undergo a net change in dipole moment as it vibrates or rotates to absorb IR radiation. e.g. HCl (Asymmetric charge distribution around molecule) -the chlorine has a higher electron density than the hydrogen. - V0 V1 is the ,fundamental absorption of molecule . -In this condition, the alternating electric field of the radiation interact with the molecule and cause changes in the amplitude(not frequency) of one of its motion or molecular vibration. -Vibrational energy change accompanied by rotational energy change. -However, the homonuclear species due to lack of net change in dipole moment are inactive. e.g N2, Cl2 etc. Modes of Vibration -It is easy to define the number & modes of vibration in di or triatomic molecules. -Hence, they can be related to energies of absorption. -It is not easy to do so in multi-atomic molecules because of many vibration centers & their interaction among several centers. -Categories of vibration a)Stretching b)Bending a)Stretching: -It is obtained by continuous changing the bond length. -As if the atoms are connected by a spring vibrating with a particular frequency. This is of 2 types: i)Symmetric(Both bond lengths are increased or decreased) ii)Asymmetric(One bond length is increased & another bond length is decreased) b)Bending: -It involves a change in the angle between two bonds. -It is of 4 types: i)Scissoring(Decrease in the bond angle) ii) Rocking(The bonds moves together in one direction) iii) Wagging(Out of plane bending together) iv)Twisting(Out of plane bending with twisting)

-No. of normal modes of vibration for non-linear a polyatomic molecule is 3n-6 , for linear molecule is 3n-5, where, n is No. of atoms. -Not all the vibrations of a molecule give rise to IR absorption. -It is because certain vibrations are inactive (e.g. in symmetric molecules) or weakly active. -A diatomic molecule can be considered as two masses connected by a spring. -A disturbance of one of these masses along the axis of the spring results in a vibration called Simple harmonic motion. -The frequency of the vibration is: = 1 . f. (m1+ m2 ) / m1. m2 S-1 2 Where, is frequency (vibration/second), f= force constant m1 & m2 are individual masses of atoms.

Instrumentation: i)Dispersive(Conventional) ii)Interferometry or FT-IR.(Advanced). 1) Dispersive instruments(Optical null & Ratio)

i)Light from source(A) is split into two equal beams. Sources:Nichrome wire, Nernst glower: globar etc.(Temp.1200- 2000)0C , 5000-7000600cm-1 ii)One beam(B) passes through the sample other behaves as reference beam. iii)Both beams are allowed to pass through the chopper(C). iv)Both beams are alternately reflected from (C) to grating monochromator(D). (v)The grating slowly rotates & individual frequencies are sent to the detector (E). Thermocouple, balometer, Golay pneumatic, Pyroelectric detectors. Infrared(thermal) energy is converted to electrical energy. vi)Upon absorption of radiation of a particular frequency by a sample, the detector alternately receives radiation of an intense beam(reference) & a weak beam(sample) . -It causes pulsating or A.C. flowing from E to F. -In case of No. absorption by sample, both the beam would have been of equal intensity. -Then, current from E to F would have been D.C. -The amplifier is designed to only to amplify A.C.

-Amplifier(F) is connencted to a small- servo-motor(G). -Upon receiving the out-of-balance signal, F causes G to drives optical wedge(H) into reference beam. -The driving continues till the E receives radiation of equal intensity from sample & reference beam. -Movement of H(attenuator) is coupled to a pen recorder(J). -Movement of H in & out of the reference beam shows as absorption bands on the printed spectrum. -The instrument balances out by optical means ,i.e. differential between the two beams -So that it is a double-beam optical Null recording spectrometer. -In expensive instruments the intensity of both the sample & reference beams can both be measured & ratioed. -This alleviates a major deficiency in optical null method(get ride of comb attenuator) . 2) Interferometry/FT-IR spectrometry. -Supersedes dispersive instrument. -It is IR spectrometry based on Michelson interferometry & Fourier-transform. - Far more superior to dispersive type by sensitivity, resolution ,speed & wavelength accuracy.

Fig.1

Fig.2

Fig.3 FT-IR (Fig.2) -IR radiation(5000-400)cm-1split into two beams. -One beam of fixed length, another of variable length (movable mirror). -When difference in the corresponding wavelength is an integer multiple of the invariable wavelength, result is constructive interference. -Destructive interference, when the difference is an odd integer multiple of one quarter of the wavelength. -The result is an interferogram. -Fourier transformation convert this time-domain interferogram into frequency domain. -Movement of mirror B is continuous & smooth. -This movement gives rise to the complete IR spectrum virtually similar with the spectrum that is generated dispersive instrument.

Sampling techniques: -Techniques depend upon state of samples: Gas, Liquid, Solid

Gases: -The gas sample is introduced into a gas cell(a). -The cell is equipped with IR transparent windows e.g. NaCl. Liquids: -A liquid sample is squeezed between two IR transparent windows (e.g NaCl flats). -Thickness of the film should be maintained 0.01-0.1mm.

-The assembled pair of the flats & liquid film are mounted in the sample beam as in (b). -Solids: Three techniques: i)KBr discs: ii)Mulls: iii)Deposited films i)KBr discs: -Prepared by grinding the sample (0.1- 2.0%) with KBr. -Then the mixture is compressed to discs. -Hard mortar & pestle are used for grinding. -The mixture is compressed to disks in special die(C).

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-Pressure about 10 tonnes load is applied. ii)Mulls(paste): -Prepared by grinding the sample with a drop of oil. -The mull is then squeezed between IR transparent windows iii)Deposited film: -Sample solution is prepared in a volatile solvent. -Solid films can be deposited onto NaCl discs after evaporation of the solvent. e.g. polymer or fatty materials. Application: i)Qualitative ii)Quantitative i)Qualitative: -All absorption bands in an IR spectrum may be considerable as a mean of identification. -Two compounds probably one if their spectra agree in all respects(position & intensity). -Identification can be done in two steps. i)Determination of functional group examining the group frequency region (3600-1250) cm-1 . ii)Involves the detail comparison of finger print region (1200-600)cm-1. Identification of impurities: -May give additional peaks than the reference spectra Study of hydrogen bonding: Shift in the spectral region, broadening of the peak -Study of polymers -Ratio of cis- trans isomers Approximate positions of some infrared abs. bands __________________________________________ Group Wave No.(cm-1 ) Wavelength(m) C-H(aliphatic) 2700-3000 3.33-3.70d C-H(aromatic Str.) 3000-3100 3.23-3.33 O-H(Phenolic) 3700 2.70 O-H(Phenolic hydrogen 3300-3700 2.70-3.03

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bonding) S-H N-H C-O C=O(Aldehyde) C=O(Ketone) C=O(Acid) C=O(Ester) C=N C-C Group C=C CC CN CH3 -, CH2C-F C-Cl C-Br C-I C-H(aromatic,out of plane bending Aromatic overtone & combination ii)Quantitative: -Based on the same underlying principle of UV/Vis. Spectrophotometry. -Analysis is possible using various techniques: i)Calibration curve of Abs. Vs.Conc. ii)Standard addition technique. iii)If Beers law is shown to be obeyed, direct comparison of sample & standard may be used. e.g . Acetazolamide, Diethyltoluamide, Activated dimethicone, phenobarbitone etc. 2570-2600 3300-3370 1000-1050 1720-1740 1705-1725 1650 1700-1750 1590-1660 3.85-3.89 2.97-3.03 9.52-10.00 5.75-5.8 5.80-5.86 6.06 5.71-5.88 6.02-6.23

Wave No.(cm-1 ) 1620-1670 2100-2250 2100-2250 1350-1480 1000-1400 600-800 500-600 500 700-610 2000-1650

Wavelength(m) 5.99-4.76 4.44-4.76 4.44-4.76 6.76-7.41 7.14-10.00 12.50-16.67 16.67-20.00 20.00

-Very less sensitive than electronic transition technique.