Spectroscopic Techniques and Applications Module 3A. INTRODUCTION The branch of physical chemistry which deals with the study of interaction of ‘electromagnetic radiations with matter is known as spectroscopy. The data obtained from these interactions isused fordetermination of molecular structure. In this section of book, we will be dealing with UV spectroscopy, IR-spectroscopy and ‘H-NMR’ spectroscopy. However before their detailed study, itisimportant to go through following important topics. Electromagnetic Radiations Electromagnetic radiation is a form of energy that is all around us and takes many forms such as X-rays, microwaves, gamma rays, etc. Sunlight is also a form of electromagnetic energy and the visible light constitute the small portion of electromagnetic spectrum. As the name indicates, these radiations have both electric and magnetic fields associated with them. Unlike sound waves and water waves, these do not require medium for their propagation i.e., these waves (e.g., light) can ravel across the great vacuum of space. These radiations are produced due to vibration of charged particle. Characteristics : 1. These radiations have dual character i.e., both particle as well as wave nature. 2. These radiations are associated with both electric and magnetic waves, oscillating perpendicular to one another as well as their direction of propagation, as shown in Fig. 3.1. Wavelength Electric flela Direction of pveceton ‘Magnetic fleld Direction of propagation Ge Fig, 3-1. Electromagnetic radiation=U, These radiations do not require any material medium for their propagation, . Ina vacuum, . ; oe all electromagnetic waves always travel at the same speed ie, (3 108 nvs), Peed of i, 5: All the electromagnetic waves are characte and velocity (v) which are related to one another ised by their wavelength (7.), freaveney 6 1 and y (wave number) = a where v is expressed in orl Cc ’ S Where v is expressed in s 6, Electromagnetic radiations are composed of photons. Each photon is associateg ,.. energy given by Planck’s equation, E = /y, where ‘is Planck’s constant. * Regions of Electromagnetic Spectrum almost the same speed ie. travel with the specs All the electromagnetic radiation: light (3 x 108 m/s“). Thus, the various electromagnetic radiations differ from one anothe; . their wave-lengths and the frequencies. “7he complete arrangement Of different types electromagnetic radiations in an increasing order of their wavelengths or decreasin, order of their frequencies is called electromagnetic spectrum.” spectrum, there are waves with the highest frequencies while at the other end are the waves ws the lowest frequencies. At the lowest frequencies, we have radiowaves. As we move» microwaves, infrared rays, visible light, UV rays and X-rays, gamma rays are encounters; followed by the highest frequency cosmic rays. Energy or Frequency ”. At the one end of ty Gamma | X-Rays uv Infrared Microwaves} Radio | Long rays Waves Rays Rays VISIBLE Wavelength Fig. 3.2. Electromagnetic spectrum The various regions of ‘electromagnetic spectrum are as follows 1. Radio frequency region (3 * 106— 3 x 10" Hz). The energy of this region is ud fr the reversal of spin ofa nucleus or electron and is thus dealt with nmr and esr spectroscopy: 2. Microwave region (3 * 10! ponds wit the energy difference between various rotational dealtwid rotational spectroscopy. 3. Infra-red region ( : the energy difference between various v' vibrational spectroscopy. —3 x 10!? Hz). The energy of this region corres I evels of the molecules and is thus +5 x 10!2—3 « 10" Hz), The energy of this region corresponds wid ibrational levels of the molecules and is thus dealt"{Visible and ultraviolet region @ x 10" — 3 x 1016 Fiz). The energy of this region corresponds to the energy difference between various energy levels of the molecules and is thus dealt with electronic spectroscopy, 5, X-ray region (3 x 10'S —3 x 10!8 Hy), This deals with the energy changes involving inner electrons ofan atom ora molecule, 6. y-ray region (3 x 10!8_ 3 x 100 Hz). This deals with the energy changes involving rearrangement of nuclear particles, types of Spectroscopy : 1. Emission Spectroscopy : The electrons of atoms or molecules ofa chemical substance undergo transition from ground state to excited state when subjected to jntense heat or electric discharge. Since the excited state is unstable, so they return to lower stable state by emitting the absorbed radiation in the form of light. On passing the emitted radiation through prism and then allowed to fall on photographic plate, emission spectrum is obtained in the form of bright lines upon a dark background. 2.Absorption Spectroscopy : Itstudies the absorption of electromagnetic radiation depending upon the molecular structure. On passing a beam of electromagnetic radiations: through asubstance, theradiations may either be absorbed or transmitted. depending upon the wavelength oftheradiation and the molecular structure. Absorption of radiation leads to the increase in the energy of the molecule which ultimately leads to electronic excitation. The absorption of radiation is observed instrumentally by passage of radiation through a sample of substance and the analysis of the transmitted light gives the absorption spectrum. The field of absorption spectroscopy can be divided into the following types, depending upon the nature of electromagnetic radiations : (a) UV-Visible (6) IR (c) Microwave (d) Nuclear magnetic resonance (e) Electron-spin resonance 3.2, PRINCIPLE OF ULTRAVIOLET-VISIBLE SPECTROSCOPY Itisalsoreferred as electronic spectroscopy. Itis used primarily to measure the multiple bonds or aromatic conjugation within molecules. UV-visiblespectroscopy requires electromagneticradiations ofhigh energy. The UV-region extends from 100—400: nm while the visibleregion extends from400 -800nm. The UV-region is further divided into near UV (200-400 nm) and far UV (100-200 nm). The region below 200 nm is named as vacuum UV because the molecules of air (O,N etc.) absorb radiation in this region, thus this region can be studied only by using evacuated instruments. Principle : When electromagnetic radiations falling in the UV-visible region are passed through the solution of organic compounds, the radiations are either absorbed or transmitted depending on the wavelength of radiation and the structure of compound. The absorption of light results in the transition of electron from lower electronic ground state to higher electronic excited state, Since it involves electron-excitation process, thus UV is sometimes called electronic Spectroscopy.Wh inti the molecules of compounds, photon-electron col tak ac a eri eee matches with the energy difference betieg a . a . Elect ieee and the excited state, electronic transition takes place, ie, thy aE =E, ~ Ey =hv= He where, ‘AE? is the energy difference between ground state and excited state. E) = Energy of ground state E, = Energy of excited state ‘v’ = frequency of radiation 3.3. ABSORPTION LAWS The absorption of light by molecules is governed by the absorption laws, discussed as follows. 1. Beer’s Law : It states that the amount of light absorbed is directly Proportional to the molar concentration of the absorbing substance, , 2 Lambert’s Law : It states that the fraction of | light absorbed by a homogeneous Medium is directly proportional to the path length of the sample of the absorbing substance. 3. Beer’s Lambert Law: This law i ofabsorbing substance. Mathematically, log Me C Beer’s law) Jog Het (Lambert’s law) | Combining Beer’s and Lambert’s law, we have | rf log 1c Cl og XC I, log 0 = ec} log P= 8 where, Ig = Intensity of incident light I = Intensity of transmitted light C = Conc. of the absorbing substance in mol! 1 = path length of sample € = molar absorptivity or molar extinctions coefficient or extinction Coefficient log Ip/I — absorbance (A) of the solution or optical density.‘A’ is the measure of absorption of radiation by a sample having values between 0-2. Thus, Beer-Lambert law may also be expressed mathematically as : A =log a =eCl itations Li Itis strictly obeyed while dealing with the UV spectra of single species. It fails in the following cases : () Sample containing different structural forms (ce, Keto-enol) in equilibrium. (i) Solutions where solute (sample) and solvent interact with one another to form complexes. (iii) Fluorescent compounds. Molar Absorptivity _—_ee Molar absorptivity (e), also known as molar extinction coefficient, is a measure of how well achemical species absorbs a given wavelength of light. It is the characteristic property of a compound and is quite independent of its concentration or the path length of light. Its value ranges from zero to 10°. The values above 104 are termed high intensity absorptions and the values below 10° are termed low intensity absorptions. Mathematically, log #2 = 2.c on log Ig/I Cl or eee ee Cl (mole Lt!) x (cm) A . 2 or 6 = = £21000 em" (= L= 1000 em3) xem mole 1000 cm? Thus, the units of molar absorptivity = 1000 cm?/mole or Area/mole But these units are never used conventionally. 3.4. BAND NATURE OF UV-SPECTRUM Absorbance of UV radiations by a molecule results in various types of electronic transitions, €8. xn",n—2',n—+0" etc. The radiations absorbed by the molecules appear as bands in the spectrum rather than peaks. The appearance of bands rather than peaks is due to the reason that electronic transitionsare always associated with vibrational and rotational transitions because of which broader; 7 peaks takes place which results in the formation of bands. SS E, (Electronic level) ¥ Enid sat Rotational levels within a vibrational level v \ Vibrational levels within 7 Hlectconic level Vo Fig. 3.3. Rotational and vibrational levels within electronic levels Each electronic level (E) has many vibrational levels (Vo, Vj, V2» V3 etc.) and each vibration level has many rotational levels in it. The electronic transition always results in change in vibrational as well as rotational energy levels. Thus a variety of wavelengths are absorbed 2. which electronic transition takes place e.g. Ep to E,. Thus band rather than a sharp peak is obtained in UV-spectrum. 3.5. TYPES OF ELECTRONIC TRANSITIONS E, (Electronic level) Ground state On the absorption of energy by a molecule in the UV region, changes are produced in the electronic energy of the molecule. This is due to the transition of electrons from bonding or non. bonding molecular orbitals (c, n, ot 2) to the next higher energy orbital i.e., an anti-bonding molecular orbital (o* or *). g The various electronic energy levels are diagrammatically represented as in Fig 3.6, along with the possible electronic transition. is Ig o* —— nt n-po* abot npr |non* ‘Antibonding orbitals Energy Non-Bonding Ronding orbitals 6 Fig 34, Electronic enery levels and elecionie tansitiong- ceteornnt , | Theelectronie transitions occurring in UV Worc* Q)nso* region of the following type: @aor not t involves the transition of cl TRY antibonding sigma (o*) orbital. Iti ma bond is very stron; » therefore the (1) o> 6* Transitions orbital to the higher ener alkanes. Since {ron from bonding sigma (0) he only type of transition seen in short wavelengths ved in the vacuum or far ultraviolet region (~ 150 nm). . methane absorbs at 125 nm, propane absorbs at 135 nm while eyelopropane absorbs at 195 nm due to the presence of, strain which raises the energy of its ground state. @) 71> ot Transitions ¢ 11 involves the transition of non-bonding electrons (p tone pair of electrons in hetero atoms such as NO, S, halogens, ele.) to the higher energy antibonding o* molecular orbital. These transitior require lower energy than o> o* trans and hence oecur at longer wavelength than a> transitions. It has been found thatthe energy eauited for thls tnsition depends on the electronegativity ofthe helero-atom. Higher the electron igher will be the frequency or energy required for n> o* tansigons. &8- (D CH3C1 absorbs at iyyq. 173 nm while CHL absorbs at max 258 nm. This difference can be explained by considering the large electronegativity of Cl than | Gi) CH5OH absorbs at 195 nm while (CH. )3 N absorbs at 227 nm. (3) 5 =* Transitions : It involves the citation of x-electron to an antibonding x* MO. This type of transitions are seen in unsaturated hydrocarbons (i.e., alkenes and alkynes), aromatic compounds, carbonyl compounds, etc. These transitions occur at less energy than n> s* tansitions. The intensity of such transitions is usually very high (ice, large value of Exnay)> eg., (i) In saturated aldehydes and ketones, > n* transition occurs at about Amay 185 nm. (i Inethene, x > x* transition occurs at 175 nm. tivity of hetero atom, his (iii) In acetylene, x— x* transition occurs at about 170 nm, (4) 2 >=* Transitions : Itinvolves the excitation of unshared electron pair or non-bonding electrons present on hetero atoms to an antibonding x* MO. Itrequires the least amount o fenergy than all the other types of transition and thus occurs at long wavelengths. These transitions are forbidden by symmetry cosiderations, thus the intensity of such transitions are very low (i.e, small value of yay 15)- e.g., In saturated aliphatic ketones, 7 — n* transition occurs at about 280 nm. 3.5.1. Concept of Chromophore and Auxochrome Chromophore. The term ‘chromophore’ was originally applied to system having functional group which imparts colour to the system or compound, For example, nitro group is a chromophore since it imparts yellow colour to the compound.aS en now-a-days, the term ‘chromophore’ is used in broader sense. Chromophore jg Sy t as any group that absorbs electromagnetic radiations in the visible or ultraviolet x, ed may or may not impart colour to the compound. a Som« i i it i foe e of the important simple chromophoric groups along With Amax ANd eax are listed, Table 3.1. Some Simple Chromophoric Groups Chromophore ‘Amax (2m) Emax Enylenic Sc=cC 175 (n> n*) 15,000 Acetylenic, —C = C— 170 (n> n*) 10,000 Carbonyl, Sexo 165 (n +0*) 5,000 190 (n->7*) 16,000 280 (n >n*) 15 oH Carboxylic —C=O 208 (n > 2*) 32 OR Ester, -¢=0 211 (a >x4) 57 Nitro, —NO, 201” > 0%) 5,000 274(n +2") 7 Az0, -N=N— 338 (n> n*) 4 In general, chromophores are classified into two types : (a) Chromophores containing multiple bonds such as Se=cd’ and —C2C— which undergo * 5 n* transitions. (d) Chromophores containing multiple bondsas well as lone pair of electrons such: as >C=0. rg0 bothx>n* and > n* transitions, _N—which unde! F 2N—wh the same functional groups generally absorb at nearly same wavelength having, ee ‘Compounds value Of &yax 5 but the position of ‘ J ye same me i max and shi (Aman) with at attaching auxoehromes oF by conjugation to chrom a 40—cH,_Np, Due to eee the energy difference between the ground and excited state of the group reases, a8 a result, the absorption shifts to longer wavelength (i.c., bathochromic shift) with spe simultaneous increase in the intensity of the absorption band. For example, an auxochrome —NH) when attached to benzene ring (chromophore) shifts be Apa longer wavelength with the simultaneous increase in Epyays Le. oa 9 Aniline shifts t Amax=255nm SSSI y= 280nm (Benzene) Amitine) shifts to Emax = 203 Sc = 1430 3.6. SHIFT OF BANDS WITH SOLVENTS, DOUBLE BONDS (CONJUGATION), CARBONYL COMPOUNDS AND AROMATIC COMPOUNDS Chromophores such as Seac’ —C=#C,-Cen, Se-o, ete. show sorption below 220 nm i.e., in far ultraviolet region which cannot be easily studied using commercially available spectrophotometers. The position of absorption maximum (i.e. Linay) aad the intensity of absorption (i.e. eax) can be modified by substitution of some groups or by ‘te change of solvent, as discussed below : 1. Bathochromic or Red shift : It involves the shift of absorption maximum towards the nger wavelength. It can be brought about by one of the following methods :cen See AI di] (@) By attaching an auxochrome to chromophore. When an auxochrome such -° : OH, or —NUy is attached to chromophore ( Sc=cX or benene ring), extends Conjugag of chromophore by sharing its non-bonding pair of electrons. ‘This extended conjugation the decrease in the energy difference between the ground and excited state and heng, max towards longer wavelength, a. For example, Benzene absorbs at Ajyay 255 nm while the substitution of hydrog benzene by —NHj leads to bathochromie shi, ic. aniline absorbs at Ang 280 nm, (6) By conjugation of two or more nophores. Bathochromic shift is also obser, When two or more chromophores are present in conjugation in a molecule. For example ethylene absorbs at Ay, 170 nm while 1, 3-buladiene absorbs at Apyay 217 nm. : (c) By change of solvent. Generally, change in the polarity of the solvent causes the significant shift in the absorption maximum (Amax) Of polar compounds only. For example Acetone shows. Amax at 264-5 nm in water and 270, nm in hexane. Thus, the use of solvent of lower polarity produces a red shift in the 7 — x* absorption, of carbonyl compounds. leads © shiny en of 2. Hypsochromic or Blue shift. It involves the shift of absorption maximum towards the shorter wavelength. It can be brought about one of the following ways : \ a @ By attaching an auxochrome to the c=0 by an auxochrome like OH, Cl, NH; etc. shifts the — n* transitions towards shorter wavelength. C=O group : Substitution of H-atom of H, O O 0 2 Z A Z H3C—C. H,C—C HC—C H;C—C Na et a ONES nee 28. Acetaldehyde Acetyl chloride Acetamide Acetic acid Day 293 0 may 235 nm AK pay 214 nim pay 208 am This shift is because of I-effect of the hetero atom which causes the lone pair of electrons present on carbonyl oxygen atom to get more tightly held by it, as a result the energy of orbital decreases more than that of x* and thus the energy gap between n and x* orbital increases and hence the absorption shifts towards the shorter wavelength. (& By increasing the polarity of the solvent. Increase in the polarity of the solvent sit’ n-— x* transition to shorter wavelength. This is duc to the greater stabil state i.¢., n-orbital through dipole-dipole interaction, hydrogen bonding or solvation. Asa rest the energy gap between n> x* orbital increases and hence absorption shifts to shorter waveleng , Acetone absorbs at 279 nm in hexane while 270 nm in ethanol and 264 nm in wale egree ~~ (¢) By removing conjugation in a system. For example, Aniline absorbs at Apa 280 nm, nd its protonation gives anilinium ion (where N has no lone pairs present on it which can ugate with benzene ring) which absorbs at 4,9, 203 nm. 3, Hyperchromic effect. It involves the increase in the intensity of absorption. It is usually seen in bathochromic shift. For example, the introduction of methyl group in position 2 of yridine iNCTEASCS trax from 2750 to 3650. 4, Hypsochromic effect. It involves the decrease in the intensity of absorption. It is usually «een in hypochromic effects. For example, the introduction of methyl group in position 2 of piphenyl distorts the geometry of biphenyl, as a result ey, decreases from 19000 to 10250. . Effect of Solvents Solvent plays an important role in UV spectroscopy. The solvent selected should be the one - that does not itself get absorbed in the region under investigation. A solvent should be transparent ina particular region. The most commonly used solvents are follows : Solvent A, for absorption Water 191 nm. Ether 215 nm Methanol 203 nm Ethanol 204 nm Chloroform 237nm Carbontetrachloride 265 nm Benzene 280 nm Tetrahydrofuran 220 nm Some of the solvent effects are as discussed below : 1. Due to the solvation, the fine structure of the absorption bands is blurred out and as a result a smooth curve is obtained. 2. The change in polarity of the solvent has no significant effect on the absorption of non- polar compounds such as dienes, aromatic hydrocarbons, etc. 3. The change in the polarity of the solvent usually shift the position of absorption band of polar solvents. Two types of shifts are usually observed in polar compounds. These are as follows : () n> x* and n — o* transitions : Increase in the polarity of the solvent shifts n > 2* transitions to shorter wavelength (i.e. blue shift). This shift is because of the greater stabilisation ofthe ground state than the excited state through dipole-dipole interaction, hydrogen bonding or Solvation with polar solvent. As a result, the energy difference between the ground state and the &xcited state increases and thus the absorption shifts to shorter wavelength. For example, Acetone absorbs at 279 nm in hexane, 270 nm in ethanol and 264 nm in water.gic i spe a" AE, 2 n—Y— Non-polar ~ solvent a n Polar solvent AE2 > AE, Fig, 3.5. Effect of polarity of solvent on 1 —> 7 transition. Similarly, the polarity of solvent shifts the 1 o* transition towards shorter wavelength, (ii) x x* transitions: Increase in the polarity of the solvent shifts x x* transition to long, wavelength (i.e. red shift). This shift is because of the, greater stabilisation of the excited state tha, the ground state through dipole-dipole interactionsand H-bonding with theexcited state. Asaresyh the energy gap between the ground and the excited state decreases and the absorption shifts toward; longer wavelength. For example, in ethene, the ground state is non-polar due to equal distribution of electrons between the carbon atoms. But in the excited state, molecule becomes polar due to the shifting of electrons towards one of the carbon atom, i.e. Coe Nb bbe dSe=cd >Sc= cd Polar Non-polar Excited state Ground state ‘Asa result, x x* transitions of ethene shifts to longer wavelength in polar solvent. Effect of Double Bonds Conjugation ‘on is a mechanism to delocalise electrons i.e., a way by which one end of the th the other chemically. Conjugation of double bond with anothe" double bond or a C=O group causes the absorption bands to shift to longer wavelength with greater intensity (i.€., &max)- FOr example, ethylene absorbs at about 175 nm while butadien® absorbs at about 217 nm ‘and trans-1, 3, 5-hexatriene absorbs at 256 nm. Thus, each double bond extending the conjugation causes the absorption lo occur at longer wavelength by about 40 nm in the lower polyenes. Explanation Epa Ethylene is composed 3. Conjugati molecule may communicate wil ofsigma bond, formed by overlap of yp? hybrid orbitals and x-bont. formed by the Tateral overlap of unbybridise 2p- aa ls, n-bond comprises of x-bonding at x*-antibonding molecular orbit ee a ence in energies between them. Due @ large energy separation, a transition In ethene takes place by absorption of radiations # short wavelength (4 below 200 nm) falling in fir UV region, very— : E a dee, jugation / In butadiene, there are two x-bonds in conjugation with one another. Extended eee so the interaction of two ethylenic units with one another and thus resulting in the form: ; a . a . + 0. apt MO’s ie., x and x (bonding) while x} and x} (antibonding) as shown in the figure 3 mE x LUMO ene LUMO nt 175 nm 217 nm mt CH=CH, ma Ethene H,C=CH—CH=CH, Butadiene Fig. 3.6. Molecular orbitals of ethene and butadiene Iris clear from the figure 3.4 that conjugation increases the energy of HOMO (i.e. 7) and decreases the energy LUMO (e., x3). Asa result, the energy level involved in x—+x* transitions (i.e., z,anda3*) come closer and thus this transition can occur at less energy ie., at longer wavelength (217nm)than the corresponding x—>x* transition in ethylene (175 nm). Onadding more conjugated double bonds to a molecule, the energy required to reach the first excited state further decreases, and thus shifting the n> x* transition to more longer wavelength. This isthe reason that 1, 3, S-hexatriene absorbs at 2, 258 nm and 1, 3, 5, 7-octatetraene absorbs Bigg 290M. Sufficient conjugation shifts the absorption to such longer wavelengths falling in the ‘visibleregion ofthe spectrum, thus resulting in colour in compounds with sufficient conjugation. Forexample, lycopene (red-pigment of tomatoes) and trans f-carotene (an orange pigment of carrot) have eleven conjugated double bonds. Ithas been found that stereostructural features may effect the absorption ofa conjugated diene. Forexample, in stilbene, the E-isomer absorbs at longer wavelength with greater intensity than the Corresponding Z-isomer. This can be explained by taking into consideration, the concept of Coplanarity which is essential for the effective: overlap of x-orbitals so that x>2* transition can occur with increased ease,