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JSPM’s

Charak College of Pharmacy & Research,

GAT No.720/1&2,WAGHOLI, Pune-NAGAR ROAD,PUNE-412 207

JSPM’s Charak College of Pharmacy & Research, GAT No.720/1&2,WAGHOLI, Pune-NAGAR ROAD,PUNE-412 207 Theory of U.V Spectrophotometry

Theory of U.V Spectrophotometry

Presented By,

Miss. Pranita Kore

Guided By,

Dr. Rajesh J Oswal Prof.Sandip Kshirsagar

Department Of Pharmaceutical Chemistry

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 Introdution  Electronic Transition’s  Chromophores  Auxochromes  Beer’s -Lambert Law 5/13/2014
  • Introdution

  • Electronic Transition’s

  • Chromophores

  • Auxochromes

  • Beer’s-Lambert Law

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I.Introduction

A.UV radiation and Electronic Excitations:

1.The difference in energy between molecular bonding, non-bonding and anti-bonding orbitals ranges from 125-650 kJ/mole

2.This energy corresponds to EM radiation in the ultraviolet (UV) region,

100-350 nm, and visible (VIS) regions 350-700 nm of the spectrum

3.For comparison, recall the EM spectrum:

g-rays

X-rays

UV

IR

Microwave

Radio

I.Introduction A.UV radiation and Electronic Excitations: 1.The difference in energy between molecular bonding, non-bonding and anti-bonding
I.Introduction A.UV radiation and Electronic Excitations: 1.The difference in energy between molecular bonding, non-bonding and anti-bonding
Visible
Visible

4.Using IR we observed vibrational transitions with energies of 8-40

kJ/mol at wavelengths of 2500-15,000 nm

5.For purposes of our discussion, we will refer to UV and VIS spectroscopy as UV

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Introduction

  • B. Band Structure

6. When these energy

levels are superimposed, the effect can be

readily

explained

any transition has the possibility of

being

observed

Energy

E 1

E 0

R 1 - R n V 4 R 1 - R n V 3 R 1
R 1 - R n
V
4
R 1 - R n
V
3
R 1 -
R n
V
2
R 1 -
R n
V
1
R 1 - R n
V o

Disassociation

R 1 - R n V 4 R 1 - R n V 3 R 1
R 1 - R n
V
4
R 1 - R n
V
3
R 1 -
R n
V
2
R 1 - R n
V
1
R 1 - R n
V
o

Disassociation

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II. Electronic transitions

The absorption of UV or visible radiation corresponds to the excitation of outer electrons.

There

are

two

types

of

electronic

transition

which

can

be

considered;

1.Transitions involving p, s, and n electrons 2.Transitions involving charge-transfer electrons

When an atom or molecule absorbs energy, electrons are promoted from

their ground state to an excited state.

In a molecule, the atoms can rotate and vibrate with respect to each other. These vibrations and rotations also have discrete energy levels, which can be

considered as being packed on top of each electronic level.

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Absorbing species containing p, s, and n electrons

Absorption of ultraviolet and visible radiation in organic molecules is restricted to certain functional groups (chromophores) that contain valence electrons of low excitation energy.

The

spectrum of

a

molecule containing these chromophores is

complex.

 

This

is

because

the

superposition

of

rotational

and

vibrational

transitions on the overlapping lines.

electronic transitions gives a

combination of

This appears as a continuous absorption band.

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Possible electronic transitios of p, s, and n electrons are;

Possible electronic transitios of p , s , and n electrons are; 5/13/2014

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s -s * Transitions

An electron in a bonding s orbital is excited to the corresponding antibonding orbital. The energy required is large.

For example, methane (which has only C-H

bonds,

and

can

only

undergo s -s * transitions) shows an absorbance maximum at 125 nm. Absorption maxima due to s - s * transitions are not seen in typical UV- Vis. spectra (200 - 700 nm)

n -s * Transitions

Saturated compounds containing atoms with lone pairs (non-bonding electrons) are capable of n - s * transitions.

These transitions usually need less energy than s - s * transitions. They can be initiated by light whose wavelength is in the range 150 - 250

nm. The number of organic functional groups with n - s * peaks in the UV region is small.

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n - p * and p - p * Transitions

Most absorption spectroscopy of organic compounds is based on transitions of n or p electrons to the p * excited state.

This is because the absorption peaks for these transitions fall in an experimentally convenient region of the spectrum (200 - 700 nm).

These transitions need an unsaturated group provide the p electrons.

in

the

molecule to

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III.Chromophores

A.

Definition

1.

Remember the

electrons present in organic molecules are

involved in covalent bonds or lone pairs of electrons on atoms

such as O or N

  • 2. Since similar functional groups will have electrons capable of

discrete classes of transitions, the characteristic energy of these energies is more representative of the functional group than the electrons themselves

  • 3. A functional group capable of having characteristic electronic

transitions is called a chromophore (color loving)

  • 4. Structural or electronic changes in the chromophore can be

quantified and used to predict shifts in the observed electronic

transitions

III.Chromophores

  • B. Substituent Effects:

General from our brief study of these general chromophores, only the weak n p* transition occurs in the routinely observed UV

The attachment of substituent groups (other than H) can shift the energy of the transition

Substituents that increase the intensity and often wavelength of an absorption are called auxochromes

Common auxochromes include alkyl, hydroxyl, alkoxy and amino groups and the halogens

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III.Chromophores

  • C. Substituent Effects

General Substituents may have any of four effects on a chromophore

i.

Bathochromic shift (red shift) a shift to longer l; lower energy

ii.

Hypsochromic shift (blue shift) shift to shorter l; higher energy

iii.

Hyperchromic effect an increase in intensity

iv.

Hypochromic effect a decrease in intensity

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e

Hypochromic Hypsochromic Bathochromic 200 nm 700 nm Fig: Absorption & intensity shifts Hyperchromic
Hypochromic
Hypsochromic
Bathochromic
200 nm
700 nm
Fig: Absorption & intensity shifts
Hyperchromic

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IV. Auxochromes

The color of a molecule may be intensified by groups called auxochromes which generally do

not absorb significantly in the 200-800nm region, but will affect the spectrum of the

chromophore to which it is attached.

The most important auxochromic groups are OH, NH2, CH3 and NO2 and their properties are acidic (phenolic) or basic.

The actual effect of an auxochrome on a chromophore depends

on

the

polarity of the

auxochrome, e.g. groups like CH3-,CH3CH2- and Cl- have very little effect, usually a small

red shift of 5-10nm. Other groups such as -NH2 and -NO2

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V. Beer-Lambert law

slit

cuvette

source

V. Beer-Lambert law slit cuvette source detector 5/13/2014
V. Beer-Lambert law slit cuvette source detector 5/13/2014

detector

V. Beer-Lambert law slit cuvette source detector 5/13/2014
V. Beer-Lambert law slit cuvette source detector 5/13/2014

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The method

is

most often

used

in

a

quantitative way to determine

concentrations of an absorbing species in solution, using the Beer-Lambert law:

A=log10 (I0/I)=ε.C.L

Where, A is the measured absorbance,

I 0 is the intensity of the incident light at a given wavelength,

I is the transmitted intensity, L the pathlength through the sample, and

c the concentration of the absorbing species.

For each species and wavelength, ε absorptivity or extinction coefficient.

is

a

constant

known as

the molar

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