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UV VIS Spectroscopy

PHRM 309

UV VIS Spectroscopy
Spectroscopy is the study of the interaction between matter and radiated energy (light).

Electromagnetic radiation (light)


Electromagnetic radiation (light) is one form of energy.

Produced by fluctuation of electric charge and magnetic field.


Electromagnetic radiation may be propagation of the energy in the harmonic wave which is produced electric charge and magnetic field atom. defined as the form of simple by oscillation of residing on the

Electromagnetic radiation is a self-propagating wave in space with electric and magnetic components. EM radiation has an electric and magnetic field component which oscillate in phase perpendicular to each other and to the direction of energy propagation.

Radio waves have longest wavelength and Gamma rays have shortest! Since electromagnetic radiation (light) is a wave, it can be characterized by either a wavelength or frequency or wave numbers.

The electromagnetic wave consists of two fluctuating fieldsone electric and the other magnetic. The two vectors are at right angles to one another, and both are perpendicular to the direction of travel.

Wavelength ()
The distance between two consecutive peaks or troughs in a particular wave. Linear distance from any point on one wave to the corresponding point on the adjacent wave.
Increasing wavelength

1 > 2 > 3
Symbol: lambda () Unit: meter (m), practically measured in micrometers (m, a millionth of a meter) or nanometers (nm, a billionth of a meter )

Frequency ()
The number of waves which can pass through a given point in one second.
Decreasing frequency 1 < 2 < 3
Symbol: nu () Units: cycle/sec, may also be denoted as hertz (Hz)

Greater the wavelength smaller the frequency.


1 Frequency Wavelength Frequency, = C/ where C is the velocity of the light, 3.00x108 msAll types of radiations travel with the same velocity and no medium is required for their propagation. They can travel through vacuum.

Wave number
Wave number is defined as the total number of waves passing through a space of unit length (1cm or 1m). We can calculate the wave number by taking the reciprocal value of wave length of a radiation. i.e.
1 Wave number = Wavelength

Energy
Energy of a particular wave of the particular radiation can also be calculated by applying the relation:
E=h. = h. (c/) Here h = Plancks constant = 6.626x10-27 erg sec = Frequency of radiation c = Velocity of EM radiation = Wavelength

Electromagnetic spectrum
The arrangement of all types of electromagnetic radiations in order of their increasing wavelength or decreasing frequencies is known as complete electromagnetic spectrum. The full electromagnetic radiation spectrum is continuous and each region merges slowly into the next. For convenience of reference, definitions of the various spectral regions have been set by the Joint Committee on Nomenclature in Applied Spectroscopy:

Fig: Electromagnetic spectrum

Region
Far ultraviolet Near ultraviolet Visible Near infrared Middle infrared Far infrared Microwave

Wavelength (nm)
10-200 200-380 380-780 780-3000 3000-30,000 30,000-300,000 300,000-1,000,000,000

Absorption spectroscopy
It may be defined as the analysis of chemical substance by the measurement of the amount of radiation absorbed by this substance. Absorption spectroscopy refers to a range of techniques employing the interaction of electromagnetic radiation (EMR) with matter.
In absorption spectroscopy, the intensity of a beam of light measured before and after interaction with a sample is compared.

Absorption spectroscopy is widely used for both qualitative (is a chemical present?) and quantitative (how much?) and structural (is it degraded?) work in a wide range of fields.

Examples of absorption spectroscopy


Infrared spectroscopy Near infrared spectroscopy Microwave spectroscopy X-ray spectroscopy (emission spectroscopy) UV-visible spectroscopy NMR spectroscopy

Vacuum ultraviolet region


The UV light below 200nm is relatively uninformative and difficult to measure. Since air also absorb radiation in this region. In order to measure absorption of UV radiation below 200nm, air must be removed from the instrument. This region is called vacuum UV region. It can excites sigma bond electrons.
The UV light above 200nm excites pi bond electrons and the region give informative spectra and can be easily measured.

Kinds of electron involved

3 kinds of electrons in organic molecules are involved


UV or Visible spectroscopy- Bonding electron (ground level)

- Non-bonding electron (ground level)


- Anti-bonding electron (excited level) Electrons of ground states are in outer most shell or valence electrons.

While two atoms form chemical bond, their atomic orbital combine together to form molecular orbital. Bonding orbital and antibonding orbital

Bonding orbital energy level is always lower than that of the original atomic orbital
Antibonding orbital energy - higher

Bonding electron
Electrons which are involved in bond formation. They are present in the lower energy level in the molecule. They are of two types: -bond electrons (electrons in the single bond, electrons in saturated compounds)

-bond electrons (electrons in double and triple bond, electrons in unsaturated compounds)

Non-bonding electron
Paired electrons in outer most shell which are not involved in bond formation between atoms in molecule. They are present in the higher energy level in the molecule than bonding electrons. They are represented by n.

Organic compounds halogen, sulphur.

contains

nitrogen,

oxygen,

Anti-bonding electron
Electrons which are excited to a higher energy level by absorbing energy (EMR) are called anti-bonding electron. They are present in the higher energy level. They are represented by: Anti-bonding sigma (*) and

Anti-bonding pi (*)

*
Energy

Vacuum UV or Far UV (<190 nm )

UV/VIS

Electron transitions

UV-VIS spectroscopy
This is the spectroscopy. earliest method of molecular

A phenomenon of interaction of molecules with ultraviolet and visible lights.

Absorption of EMR results in electronic transition of a molecule, and electrons are promoted from ground state to higher electronic states.

The radiation which is absorbed has an energy which exactly matches the energy difference between the ground state and the excited state. These absorptions transitions. correspond to electronic

At room temperature, most of the atoms, molecules and electrons are in the lowest energy orbital called ground state. The electron of atom (molecule) at ground state can absorb EMR and transit to higher energy orbital called excited state.

Atom or molecule can absorb the radiation only when the energy of proton is equal to the energy difference of the two orbitals.

* Transitions
An electron in a bonding 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 * transitions) shows an absorbance maximum at 125 nm. Absorption maxima due to * transitions are not seen in typical UV-VIS spectra.

n * Transitions
Saturated compounds containing atoms with lone pairs (non-bonding electrons) are capable of n* transitions. These transitions usually need less energy than * transitions. They can be initiated by light whose wavelength is in the range 150 - 250 nm. The number of organic functional groups with n* peaks in the UV region is small.

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

s* p*
Atomic orbital

Unoccupied levels

Energy

Atomic orbital

Occupied levels

p s
Molecular orbitals

pp*

max(Lambda max): For a given substance, the wave length at which maximum absorbance in the electromagnetic spectrum occurs is known as max.

Solvent selection criteria


Diluted solution is used for analysis- so solvents has great influence on the process. The criteria are: Should not absorb radiation (%T=100) Should be less polar- to avoid interaction Solvents do not contain double bonds are most suitable Should be free from impurities.

Solvent selection criteria


Most commonly used solvent is 95% ethanol. Commercial grade solvent contain benzene as impurity.

Solvent

At which or above which remain transparent


205 nm 210 nm

Water Methanol, Ethanol, Ether, Cyclohexane, n-hexane

Chloroform Benzene

245 nm 280 nm

Transition of electron of acetone (n *)


CH3 C = CH3

Polarity of solvent decrease


Solvents
max

H2O 264.5

CH3OH 270

C2H5OH 272

CHCl3 277

C6H14 279

(nm)

Polarity
In chemistry, polarity refers to the dipole-dipole intermolecular forces between the slightly positivelycharged end of one molecule to the negative end of another or the same molecule.
Molecular polarity is dependent on the difference in electronegativity between atoms in a compound and the asymmetry of the compound's structure. For example, a molecule of water is polar because of the unequal sharing of its electrons in a "bent" structure, whereas methane is considered non-polar because the carbon shares the hydrogen atoms uniformly.

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


Hypsochromic shift (blue shift) shift to shorter ; higher energy Hyperchromic effect an increase in intensity Hypochromic effect a decrease in intensity

Chromophores: functional groups that give electronic transitions.


Auxochromes: substituents with unshared pair e's like OH, NH, SH ..., when attached to chromophore they generally move the absorption max. to longer . Bathochromic shift: shift to longer , also called red shift. Hysochromic shift: shift to shorter , also called blue shift. Hyperchromism: increase in of a band. Hypochromism: decrease in of a band.

Hypsochromic Hypochromic

Hyperchromic Bathochromic

200 nm

700 nm

Chromophores:
Functional groups that give electronic transitions. 1. Chromophore containing electrons: ethylene

2. Chromophore containing and n electron

Auxochromes:
Substituents with unshared pair e's like OH, NH2, NHR, NR2 SH, OR, etc when attached to chromophore they generally move the max to longer and also increase the intensity of absorption. Combination of chromophore and auxochrome give rise to another chromophore Also known as color enhancing group.

Auxochrome increase conjugation of chromophore by lone pair of electron.

NH2

NH3+

Bezene Aniline Protonated Aniline max 255 nm max 280 nm max 200 nm max 203 max 1430 max ???

Biphenyl max 250 nm max 19,000


CH3

2-Methyl biphenyl max 237 nm max 10,250

max 255nm max 203

max 280nm max 1430

max 250nm max 19,000

max 237nm max 10,250

Extending conjugation has a larger effect on lmax; shift is again to longer wavelengths H H

C
H

C H

Ethylene (IUPAC name: ethene)

lmax 170 nm

H C H H H3C C H C C

H H C C H H H C lmax 263 nm conjugated triene lmax 217 nm (conjugated diene)

C
C H C

CH3

Lycopene
orange-red pigment in tomatoes

lmax 505 nm 11 conjugated double bonds

lmax 451 nm 11 conjugated double bonds

Lamberts Law
When a beam of monochromatic radiation passes through a homogenous absorbing medium, the intensity of the emitted radiation decreased exponentially as the thickness of the absorbing medium increases arithmetically.

Beers Law
When a beam of monochromatic radiation passes through a homogenous absorbing medium, the intensity of the emitted radiation decreased exponentially as the concentration of the absorbing substance increases arithmetically.

Beer-Lamberts Law
The intensity of emitted electromagnetic radiation through a sample at a given wavelength decreases exponentially with the sample thickness and concentration.

Beer-Lamberts Law
The Beer-Lambert Law is rigorously obeyed when a single species is present at relatively low concentrations.

The Beer-Lamberts Law is not obeyed:


- High concentrations - Solute and solvent form complexes - Thermal equilibria exist between the ground state and the excited state

- Fluorescent compounds are present in solution

Transmittance Transmittance is the percentage of incident light that passes through a sample of material. A= log 1/T A= absorbance T= transmittance If sample had a % transmitted light through sample of 80%. What is the absorbance of the sample? Ans: 0.096