UV-VISIBLE

SPECTROSCOPY
 SPECTROSCOPY
 It is a branch of science that deals with the study of interaction of electromagnetic radiation
with matter.
 Electromagnetic radiation consists of discrete packages of energy, called as photons.
 Photons consists of an oscillating electric field(E) & an oscillating magnetic field(M) which are
perpendicular to each other.
 Frequency(ν): It is defined as the number of times electrical field radiation oscillates in one second. The
unit for frequency is Hertz(Hz).
1Hz = 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.
As photon is subjected to energy, so
E=hν = hc/ λ
ELECTROMAGNETIC SPECTRUM
 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 or

wavelength.

 1. Absorption Spectroscopy: An analytical technique which concerns with the measurement of

absorption of electromagnetic radiation. e.g. UV(185-400nm)/ Visible(400-800nm) Spectroscopy,

IR Spectroscopy (0.76-15 μm)

 2. Emission Spectroscopy: An analytical technique in which emission is dispersed to some property

of the emission & the amount of dispersion is measured. e.g. Mass spectroscopy
 THE LAWS OF ABSORPTION

 The absorption of light by any absorbing material is governed by two laws:

 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.
-dI/dt ∝ I
-logI/I0=e.l
 -ln(I) = kt+b - (i)
 Beer’s Law: It states that ‘The intensity of a beam of monochromatic light decrease exponentially
with the increase in concentration of the absorbing substance.’
Arithmetically; -dI/dc ∝ I
I = I0.e-kc -(ii)
 The combination of (i) & (ii) gives

A = εcl
 TYPES OF TRANSITION
 Spectrometer is an instrument designed to measure the spectrum of compound

 Instrument used to measure the absorbance in UV (200-400nm) or visible (400-800nm) region is

called UV-Visible spectrophotometer

 In Uv-Visible spectroscopy molecule undergo electronic transition involving σ,π and n electrons.

 The possible transitions are

i) σ → σ* transition
ii) π → π* transition
iii) n → σ* transition
iv) n → π* transition
i) σ → σ* transition
• σ electron from orbital is excited to corresponding anti-bonding orbital σ*.
• The energy required is larger for this transition than others.
• e.g. Methane(CH4) has C-H bond only and can undergo σ → σ* transition and shows absorbance
maxima at 125 nm
 ii) π → π* transition ; 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* transition. e.g alkenes generally absorb in the region 170 to 205
nm.
 iii) n → σ* transition : 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* transition

 iv) n → π* transition : 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
 CHROMOPHORE
 The part of a molecule responsible for imparting color, are called as chromophores. 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

 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
 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.

 e.g. Benzene λmax = 255 nm, Phenol λmax = 270 nm, Aniline λmax = 280 nm
ABSORPTION & INTENSITY SHIFTS
 BATHOCHROMIC SHIFT (RED SHIFT)
 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.
 In alkaline medium, p-nitrophenol shows red shift. Because negatively charged oxygen delocalizes
more effectively than the unshared pair of electron.
 HYPSOCHROMIC SHIFT (BLUE SHIFT)
 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.
 e.g. Aniline shows blue shift in acidic medium, it loses conjugation.
 HYPERCHROMIC EFFECT
 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.

 For example, the B-band for pyridine at 257 mµ εmax 2750 is shifted to 262 mµ εmax 3560 for 2- methyl

pyridine. Introduction of an auxochrome usually increases intensity of absorption.

 HYPOCHROMIC EFFECT

 It is defined as an effect due to which the intensity of absorption maximum decreases, i.e.,

extinction coefficient, εmax .

 For example, biphenyl absorbs at 250 µm εmax 19000 whereas 2-methyl biphenyl absorbs at 237

µm, εmax 10250 (εmax decreases). Introduction of methyl group distorts the geometry of the
molecules thus, cause hypochromic effect.
 WOODWARD-FIESER RULES FOR CALCULATING
ABSORPTION MAXIMUM IN DIENES
 Woodward formulated certain empirical rules for calculating the λ max in case of dienes, later this

rules were modified by Fieser in 1948.

 According to these rules each type of diene has a certain fixed basic value and the value of

absorption maximum (λmax) depends on:

 i)The number of alkyl substituents or ring residues on the double bond.

 ii)The number of double bonds which extend conjugation and

 iii)The presnece of polar group such as –Cl, –Br, –OR, –SR etc.
 Longer the conjugated system, greater is the wavelength of absorption maximum. The intensity

of absorption (εmax) also increases with the increase in the length of chromophore. In addition,
the presence of alkyl group on the double bond causes bathochromic shift. Different types of
double bonds in conjugation are described below:

 1. Acyclic dienes or dienes contained in an open chain system where basic unit is butadiene

system.
 2. Homo-annular conjugated double bonds are the conjugated double bonds present in the same

ring. It is also called Homodiene. Examples are mentioned below:

 3. Hetero-annular conjugated double bonds are the conjugated double bonds which are not present

in the same ring. Examples are as follows:

 4. Exocyclic and endocyclic conjugated double bonds are as follows:
 Woodward’s Rule For carbonyl compounds/Enones-
 SOLVENTS USED IN UV-VISIBLE SPECTROSCOPY
 Choice of solvent is quite important.
 For the selection of a solvent, three essential requirements are-
 1) All molecules show there particular absorption bands in absorption spectrum. So, the solvent
must be transparent within the range being examined for the substance.
 Some commonly used solvents are-
 2) The effect of solvent on absorption bands- Nature of solvent greatly affect the absorption
spectrum.
 A non- polar solvent does not hydrogen-bond with the solute, and the spectrum resemble closly to its
original one.
 Whereas in a polar solvent hydrogen-bonding forma a solute-solvent complex, and a fine structure
may disappear

UV-Visible spectra of phenol in methanol and isocyanate

 3) A good solvent has ability to influence the wavelength of UV-light that will be absorbed via
stabilization of either the ground or excited state
 As the polarity of a solvent is increased, π → π* bands undergo red shift whereas n → π* blue shifts.
This is because in π → π* transition excited state is more polar than ground state in polar solvents
whereas in n → π* transition ground state with two n electrons receives greater stability than the
excited state with only 1 n electron.
 EFFECT OF SUBSTITUENTS PRESENT ON DOUBLE BOND
 When an alkyl group or ring residue is present on the olefinic carbon atoms, the absorption band is
shifted towards longer wavelength.
.  Thus, from wavelength of absorption,
di- tri- and tetra-substituted double
bonds can be distinguished
 APPLICATIONS OF U.V. SPECTROSCOPY
 Detection of Impurities - UV absorption spectroscopy is one of the best methods for determination of
impurities in organic molecules. Additional peaks can be observed due to impurities in the sample and it can
be compared with that of standard raw material. By also measuring the absorbance at specific wavelength,
the impurities can be detected.
 U.V. SPECTRA OF PARACETAMOL (PCM)
 Structure elucidation of organic compounds.- UV spectroscopy is useful in the structure elucidation

of organic molecules, the presence or absence of unsaturation, the presence of hetero atoms.  From
the location of peaks and combination of peaks, it can be concluded that whether the compound is
saturated or unsaturated, hetero atoms are present or not etc.

 Quantitative Analysis - UV absorption spectroscopy can be used for the quantitative determination of

compounds that absorb UV radiation. This determination is based on Beer’s law which is as follows.

 A = log I0 / It = log 1/ T = – log T = abc = εbc

Where : ε -is extinction co-efficient,

c- is concentration,

b- is the length of the cell that is used in UV spectrophotometer.

 Qualitative Analysis - UV absorption spectroscopy can characterize those types of compounds

which absorbs UV radiation. Identification is done by comparing the absorption spectrum with
the spectra of known compounds.
 Detection Of Functional Groups- This technique is used to detect the presence or absence of

functional group in the compound

 Absence of a band at particular wavelength regarded as an evidence for absence of particular

group
 Examination Of Polynuclear Hydrocarbons - Benzene and Polynuclear hydrocarbons have

characteristic spectra in ultraviolet and visible region. Thus identification of Polynuclear

hydrocarbons can be made by comparison with the spectra of known Polynuclear compounds.

 Polynuclear hydrocarbons are the Hydrocarbon molecule with two or more closed rings; examples

are naphthalene, C10H8 , with two benzene rings side by side, or diphenyl, (C6H5)2, with two

bondconnected benzene rings. Also known as polycyclic hydrocarbon.

 Molecular Weight Determination - Molecular weights of compounds can be measured

spectrophotometrically by preparing the suitable derivatives of these compounds.

 For example, if we want to determine the molecular weight of amine then it is converted in to

amine picrate. Then known concentration of amine picrate is dissolved in a litre of solution and its
optical density is measured at λmax 380 nm.

 After this the concentration of the solution in gm moles per litre can be calculated by using the

following formula.
 "c" can be calculated using above equation, the weight "w" of amine picrate is known. From "c"

and "w", molecular weight of amine picrate can be calculated. And the molecular weight of
picrate can be calculated using the molecular weight of amine picrate.
THANK-YOU