Spectroscopy

Spectroscopy is the study of the interaction between radiation and matter as a function of wavelength λ. It also can refer to interactions with particle radiation or to a response to an alternating field or varying frequency ν. A plot of the response as a function of wavelength or more commonly frequency is referred to as a spectrum. Spectrometry is the measurement of these responses and an instrument which performs such measurements is a spectrometer or spectrograph

Applications ::

 Physical chemistry
 Analytical chemistry  Organic and inorganic chemistry  Astronomy……..

Spectroscopy
Classifications

Nature of Excitation measured

Measurement Processes

Common Types
 Fluorescence  X-ray  Flame  Visible  Ultra-violet  Infrared  Raman  NMR  Photoemission  Mössbauer

 Electromagnetic  Electron  Mass  Dielectric

 Absorption  Emission  Scattering

Types of Spectroscopy Modern techniques for structure determination of unknown compounds include: • • • • • • • Ultraviolet (UV)-visible spectroscopy corresponds to excitations of outer shell electron between the energy levels that correspond to the molecular orbital of the systems. Infrared (IR) spectroscopy measures the bond vibration frequencies in a molecule and is used to determine the functional group. What functional groups are present in the compound. Mass spectrometry (MS) fragments the molecule and measures the masses. What is the size and formula of the compound. Nuclear magnetic resonance (NMR) spectroscopy detects signals from hydrogen atoms and can be used to distinguish isomers. What is the carbon-hydrogen framework of the compound. Fluorescence is the emission of photons from electronically excited states of atoms, molecules, and ions it is used to determine the excited state property of the molecule. MÖssbauer spectroscopy (or γ -ray spectroscopy) is concerned with transitions between energy levels within the nuclei of atom. It can be used to determine the valance state of metal in unknown compound. Raman effect may be defined as phenomenon due to which scattering of light has a slightly different frequency from that of incident light and there occurs a change in atomic oscillation within the molecule. It is helps in the elucidation of molecular structure, for locating functional groups or chemical bonds in the molecules.

Spectroscopy and the Electromagnetic Spectrum
The electromagnetic spectrum covers a continuous range of wavelengths and frequencies, from radio waves at the lowfrequency end to gamma (γ ) rays at the high-frequency end
higher frequency shorter wavelength
lower frequency longer wavelength

Spectroscopy and the Electromagnetic Spectrum .

Electromagnetic energy is transmitted in discrete amounts called quanta Amount of energy. .62 × 10 -34 J ⋅s . corresponding to 1 quantum of energy (1 photon) of frequency (ν )∈ expressed by the Planck equation is hc ∈ = hν = The energy of a photon varies directly with its frequency but inversely with its λ wavelength h = Planck's constant = 6.

E is energy of Avogadro's number ( N A ) of quanta ∈ . or.20 × 10-4 kJ/mol E= = λ λ (m) It is known as one Einstein of energy . energy of one mole of photons of wavelength λ N A hc 1.

The atmospheric absorption below 200 nm is a blessing to all. (iii) Visible region: The region from 380-780 nm A tungsten filament lamp is generally used for the visible region of the spectrum.UV-Visible Spectroscopy • Region of greatest interest to organic chemists from 2 x 10-7 m to ~8 x 10-7 meters (i) Far (or vacuum) ultraviolet: The region from 10-200 nm can be studied in evacuated systems and is termed as “vacuum ultraviolet”. since it prevents the hazardous (high energy) ultraviolet radiation in the sunlight from striking the earth’s surface. . (ii) Near or quartz ultraviolet : The region from 200-380 nm The atmosphere is transparent in this region and quartz optics may be used to scan from 200-380 nm. including the spectroscopists.

In a typical experiment. the molecules or atoms start at lower energy and go to a higher energy level upon absorption of radiation of appropriate wavelength After n io t rp so Energy Before Ab ∆ E .

After After After Energy Before . If there are several different upper levels (and there usually are) then several transitions will be observed.Absorption can only occur when the energy of the radiation (calculated from the frequency or wavelength) matches the energy gap.

Chemical compounds are coloured because they absorb visible light. UV/visible Spectroscopy In general. even organic compounds that are colourless will absorb UV light. Absorption of visible light Where has the energy that was within the photons gone to ? .

In the excitation the energy of the whole molecule increases. After n io t rp so Energy Before Ab ∆ E .In UV/visible spectroscopy the energy of the absorbed photon is used to drive the molecule into an excited electronic state.

lies in deep UV. In organic compounds which have only single bonds between the atoms the excitation energy is very high. This excitation gives a dramatic decrease in bond order due to excitation from a bonding to an anti-bonding orbital .This overall change is typically due to promotion of a single electron from a lower to higher energy orbital. The energy of the transition depends on the gap between the two orbitals.

With increasing conjugation. . the decreasing energy gap is reflected by absorption at longer wavelengths.

17-DimethoxyViolanthrone .The structures of many coloured compounds show they are very extensively conjugated. beta-Carotene HOOC COOH O H N N H N N O trans-Crocetin O O H2N Xanthopterin OMe OMe 16.

is excited from or to.Substituents added to the compound may alter the energy of the orbitals which e. Auxochromes: substituents that alter the wavelength or intensity of the absorption due to the chromophore O NH2 ORANGE O NH2 PURPLE O O NHCH3 O OH BLUE O NHCH3 .

pH indicators. Phenolphthalein O O -2H O + O O OH HO O- colourless pink .Changes in chemical composition can give rise to pronounced colour changes since this can dramatically alter the energies of the orbitals involved in the transitions e.g.

Methyl orange H3C H3C N N N red SO3- H+ H3C H3C N H N N + orange-yellow SO3- .

e.The basic laws of photochemistry: 1. 2. The Grotthuss-Draper law: Only those radiation which are absorbed by the reacting system are effective in producing chemical change. In many cases. . the system on absorbing light may or may not result into a chemical reaction. But the reverse of this law is not always true i. the absorbed light is converted into the kinetic energy of the absorbing molecules and thereby only heat effects are produced. ∆ E = NAhν It is conventionally known as one Einstein of energy. Laws of photochemical equivalence or Einstein law: Each light absorbing molecule in a photochemical reaction absorbs only one quantum of light which causes the activation.

molarity) . since A = log10 I0 /I ε is the molar absorbtivity with units of L mol-1 cm-1 b is the path length of the sample in cm c is the concentration of the compound in solution. expressed in mol L-1 (or M.The Quantitative Picture • Transmittance: • Absorbance: A = -log10 T = log10 I0/I T = I/I0 I0 (power in) I (power out) B(path through sample) • The Beer-Lambert Law: A = ε bc Where the absorbance A has no units.

10-5 M. to achieve scan Matched quartz cuvettes Sample in solution at ca.How Do UV spectrometers work? Rotates. . System protects PM tube from stray light D2 lamp-UV Tungsten lamp-Vis Double Beam makes it a difference technique Two photomultiplier inputs. differential voltage drives amplifier.

Types of Electronic Transitions Energy absorbed from UV radiation promotes an electron from highest occupied molecular orbital (HOMO) to the lowest unoccupied molecular orbital (LUMO) σ n π σ * * π n π * σ * .

700 nm) . The energy required is large. Absorption maxima due to σ − σ * transitions are not seen in typical UV-Vis spectra (200 . methane (which has only C-H bonds. For example. and can only undergo σ − σ * transitions) shows an absorbance maximum at 125 nm.σ .σ * Transitions (120-200 nm) • An electron in a bonding σ orbital is excited to the corresponding antibonding orbital.

They can be initiated by light whose wavelength is in the range 150 .n − σ * Transitions • Saturated compounds containing atoms with lone pairs (non-bonding electrons) are capable of n − σ * transitions. Methanol λ Trimethylamine λ mx a mx a = 183 nm = 227 nm ε = 500 ε = 900 . The number of organic functional groups with n − σ * peaks in the UV region is small.250 nm. These transitions usually need less energy than σ − σ * transitions.

. These transitions need an unsaturated group in the molecule to provide the π electrons. This is because the absorption peaks for these transitions fall in an experimentally convenient region of the spectrum (200 700 nm).n − π * and π − π Transitions * • Most absorption spectroscopy of organic compounds is based on transitions of n or π electrons to the π * excited state.

n − π * and π − π Transitions (continue) * • Molar absorbtivities from n − π * transitions are relatively low because the electrons in the norbital are situated perpendicular to the plane of the π bond and hence the probability of the jump of an electron is very low and range from 10 to100 L mol-1 cm-1 . . π − π * transitions normally give molar absorbtivities between 1000 and 10.000 L mol-1 cm-1 .

Benzenoid bands) from the π . єmx <100 a a Hypsochromic shift (blue shift) with an increase in solvent polarity • K band (conjugation band.The corresponding absorption band in UV-Visible spectrum may be designated as follows: • R band (group type. . This band can be used to identify aromatic compound.π * transition.π * transition. High єmx ( > 104) a Example: Dienes. Konjugierte) form π . Radikalartig) originated from n . Acetophenone B band (Benzene band.π * transition of Benzene. Broad band with fine structure between 230 – 270 nm. єmx < 100 a Example: Acetone λ mx 279 nm.

.• E band (Ethylenic bands) also from π .π * transition of ethylenic band in benzene E1 band and E2 bands of benzene occur near 180 and 200 nm respectively and their molar absorptivity varies between 2000 and 14000.

but can enhance the absorption. • • • • • . e. Bathochromic shift – red shift Hypsochromic shift – blue shift Hyperchromism – an increase in absorption Hypochromism – a decrease in absorption Isobestic Point: A point common to all curves produced in the spectra of a compound.g.Important useful terms in UV-Vis spectroscopy • • • The absorbing groups in a molecule are called chromophores A molecule containing a chromophore is called a chromogen An auxochrome does not itself absorb radiation. phenol-phenolate conversion as a function of pH can demonstrate the presence of the two species in equm by the appearance of an isobestic point in the spectrum. If an auxochrome is attached to a C=C a bathochromic effect is observed and if to a double bond where n electron are available (C=O) a hypsochromic effect is observed.

• The reverse (i. π • Peaks resulting from n − * transitions are shifted to shorter wavelengths (blue shift) with increasing solvent polarity. This arises from increased solvation of the lone pair. .Solvent effect • The solvent in which the absorbing species is dissolved also has an effect on the spectrum of the species. red shift) is seen for π − π * transitions.e. which lowers the energy of the n orbital.

generally referred as “spectrograde” A good solvent should be transparent over the desired range of wavelength.Choice of solvent: The solvent should be of high purity. Usually solvents which do not contain conjugated system are most suitable for running the Uv-visible spectrum. The fine structure of an absorption band depend on the polarity of the solvent. In a polar solvent hydrogen bonding forms a solute-solvent complex and the fine structure may disappear. A solvent should be chosen so that it does not react chemically with the sample. N O O CF3 Coumarin 153 (C-153) . A nonpolar solvent does not form hydrogen-bond with the solute and the spectrum of the Solute clearly approximate to its spectrum in the gaseous state.

Solvents for UV (showing high energy cutoffs) Water CH3C≡ N C6H12 Ether EtOH Hexane MeOH Dioxane 210 210 210 210 210 210 215 225 THF CH2Cl2 235 CHCl3 CCl4 benzene Acetone 245 265 280 330 220 Various buffers for HPLC. check before using. .

-O-(acyl) (-O-COR) S-(alkyl) -NRR+ 5 nm + 6 nm. 0 nm +30 nm +60 nm (vii) (viii) (ix) (x) .Woodward-Fieser rules for substituted dienes (i) (ii) (iii) (iv) (v) (vi) Parent acyclic diene Parent heteroannular diene Parent homoannular diene Increment for substitution Alkyl group or ring residue (if the alkyl group is attached to two double bonds. count it twice) 217 nm 214 nm 253 nm +5 nm Exocyclic double bond (effect is two-fold if bond is exocyclic +5 nm to two ring) Double bond extending conjugation substituents on vinyl +30 nm carbons (for each one) Halogen (-Cl. -Br) -OR.

.

.

.

.

Longer the conjugation. . Hence the conjugation is reduced by reduction in the π orbital overlap.The effect of steric hindrance to coplanarity Woodward rules give reliable result only for those compounds in which there is no strain around the chromophore. If in a structure the π electron system is prevented from achieving coplanarity. a This departure is due to steric crowding which distorts the geometry of the chromophore. higher will be the absorption maximum and larger will be the value of extinction coefficient. there is a marked shift in the absorption maximum and ε mx ..

when mixed.Study of charge transfer complex: The formation of charge transfer complexes occurs between molecules which. allow transfer of electronic charge through space from an electron rich molecule to an electron-deficient molecule. . The bond formation between the molecules occurs when filled π orbitals (or non-bonding orbital) in the donor overlap with the depleted orbitals in the acceptor resulting in the production of new MO. The transition between these newly formed orbitals are responsible for the new absorption band observed in the charge transfer complex. The electron transfer from the donor to the acceptor is more complete in the excited State than in the ground state and the wavelength of absorption is correlated with electron affinity of the acceptor and the ionization potential of the donor.

The basis behind this method is that the acquired absorbance should be a mixture of the host.The Benesi-Hildebrand method is a mathematical approach used in the determination of the equilibrium constant K and stoichiometry of nonbonding interactions. (G0>>H0) . then the absorbance from H0 should be negligible. the Benesi-Hildebrand method can be employed. where the guest and host are reactants. as shown below. This method has been typically used to study reaction equilibriums that form 1:1 guest-host complexes. guest. To observe one-to-one binding between a single host (H) and guest (G) using UV/Vis absorbance. and the host-guest complex. With the assumption that the initial concentration of the guest (G0) is much larger than the initial concentration of the host (H0).

A binding isotherm can be described as "the theoretical change in the concentration of one component as a function of the concentration of another component at constant temperature. This change in absorbance (ΔA) is what is experimentally acquired. with A0 being the initial absorbance before the interaction of HG and A being the absorbance taken at any point of the reaction.The absorbance can be collected before and following the formation of the HG complex. one can expect that [G] = [G]0. Δε represents the change in value between εHG and εG. Using the Beer-Lambert law." This can be described by the following equation: . the equation can be rewritten with the absorption coefficients and concentrations of each component Due to the previous assumption that [G]0 >> [H]0.

Δε can be derived from the intercept while Ka can be calculated from the slope. . Further modifications results in an equation where a double reciprocal plot can be made with 1/ΔA as a function of 1/[G]0.By substituting the binding isotherm equation into the previous equation. the equilibrium constant Ka can now be correlated to the change in absorbance due to the formation of the HG complex.

Limitation: .

Color. and the Chemistry of Vision Colored organic compounds have extended conjugated systems • “UV” absorptions extend into the visible region ∀ β -Carotene has λ max = 455 nm – When white light strikes β -carotene wavelengths in the blue region are absorbed while the yellow-orange colors are transmitted to our eyes .Conjugation.

Color.Conjugation. and the Chemistry of Vision β -carotene is converted in the human body to 11-cis-retinal. an essential molecule for vision .

Conjugation. and the Chemistry of Vision In the rod cells of the eye 11-cis-retinal is converted into rhodopsin. which is accompanied by a change in geometry The change in geometry causes a nerve impulse to be sent through the optic nerve to the brain. Color. trans-rhodopsin (metarhodopsin II) is produced. a light-sensitive substance When light strikes rhodopsin. where vision is perceived Metarhodopsin II is then recycled back into rhodopsin .

Color.Conjugation. and the Chemistry of Vision The cis-trans change in bond geometry accompanying vision .

.

π • This effect also influences n − * transitions but is overshadowed by the blue shift resulting from solvation of lone pairs. . This effect is greater for the excited state. which lower the energy levels of both the excited and unexcited states. and so the energy difference between the excited and unexcited states is slightly reduced .resulting in a small red shift. red shift) is seen for π − π * transitions. This is caused by attractive polarisation forces between the solvent and the absorber.Solvent effect (cont) • The reverse (i.e.