Spectroscopy

UV/Vis + FTIR

Dr. Muhammad Shoaib

 UV/Vis Spectroscopy
I. Introduction
A. UV/Vis radiations and Electronic Excitations
1. The difference in energy between molecular bonding, non-bonding and anti-
bonding orbitals ranges from 125-650 kJ/mole

g-rays X-rays UV IR Microwave Radio

Visible

2. This energy corresponds to EM radiation in the ultraviolet (UV) region, 200-400

nm, and visible (Vis) regions 400-800 nm of the spectrum.

3. UV/Vis spectroscopy is absorption spectroscopy in which light of UV/Vis region

is absorbed and results in the electronic transitions. 2
 UV Spectroscopy

C. Observed electronic transitions

Here is a graphical representation

s*
Unoccupied levels
p*

Energy Atomic orbital Atomic orbital

n

Occupied levels
p

s
Molecular orbitals
 UV Spectroscopy

C. Observed electronic transitions

possible electronic transitions that can occur, each of a different relative energy:

s*
s s* alkanes
p*
s p* carbonyls

p p* unsaturated cmpds.
Energy
n
n s* O, N, S, halogens

n p* carbonyls
p

s
C. Observed electronic transitions
Although the UV spectrum extends below 100 nm (high energy), oxygen in the
atmosphere is not transparent below 200 nm

Special equipment to study vacuum or far UV is required

Routine organic UV spectra are typically collected from 200-800 nm

s s* alkanes 150 nm

s p* carbonyls 170 nm

p p* unsaturated cmpds. 180 nm √ - if conjugated!

n s* O, N, S, halogens 190 nm

n p* carbonyls 300 nm √
 Lambert’s Law
When a beam of monochromatic light is passed through a substance, the
absorption of light is directly proportional to the path length of the sample.
A∝l
Beer’s Law
When a beam of monochromatic light is passed through solution,
absorption is directly proportional to the molar concentration of substance.
A∝c

Beer Lambert’s
A ∝ cl Law
A = cl
where  is molar attenuation coefficient molar absorptivity; molar extinction coefficient
 Transmittance and absorbance

A monochromatic light transmitted through a solution; with an incident intensity of I0 and

a transmitted intensity of I.
Transmittance, T, is the ratio of the transmitted intensity, I, over the incident intensity, I0.
T=I/Io
Its values is between 0 and 1 and more commonly expressed as a %transmittance.
T(%) = 100 I/Io
Absorbance, A, is related to the transmittance and incident and transmitted intensities as
A=log Io/I
A= -log T = cl
 Sample Handling
Virtually all UV spectra are recoded in solution phase

Cuvette can be made of plastic, glass or quartz

Only quartz is transparent in the full 200-700 nm range

Plastic and glass are only suitable for visible spectra

Concentration is empirically determined

A typical sample cell (commonly called a cuvette)

 Sample handling

• Sample cells are normally a square or rectangular tube with two polished ,
optically smooth faces on opposite sides parallel to each other.

• The distance between the inside faces is the optical path length (remember
Beer’s Law) and this can vary but the most popular size is 1cm.
 Sample handling

Solvents must be transparent in the region to be observed

The wavelength where a solvent is no longer transparent is referred to as the

cutoff wavelength.

Since spectra are only obtained up to 200 nm, solvents typically only need to lack

conjugated p systems or carbonyls.

 Sample handling
• Common solvents and cutoff wavelength in nm

Solvents Cutoff wavlength in nm

Acetonitrile 190 nm
cyclohexane 195 nm
chloroform 240 nm
1,4-dioxane 215 nm
95% ethanol 205 nm
N-hexane 201 nm
methanol 205 nm
Water 190 nm
Components of Spectrophotometer
 Lamps used in UV-Vis Spectrophotometer
• Tungsten Lamp
• Similar to a house hold lamp, contains a Tungsten filament
• Evacuated and filled with inert gas
• Filament can be heated up to 3000 K, above this Tungsten sublimate.
• Produce polychromatic light is required.
• Some amount of halogen is also introduced (usually Iodine)
• DEMERIT: It emits major portion in near IR region of the spectrum.

Deuterium Lamp (deuterium gas)

UV Region: Wavelength Range : 190~420nm

Xenon Lamp (xenon gas)

Wavelength Range : 190~800nm
 Monochromator
 It breaks the polychromatic radiation into component wavelengths.

 It consists of :

 Entrance slit – defines narrow beam of radiation from source.

 Collimating lens (polished surface) - collimates the lights.

 Prism (make-quartz)- disperses the light into specific wavelength.

 Focusing lens -captures the dispersed light & sharpens the same to the sample cuvette
via exit slit

 Exit slit- allows the corrected wavelength of light to the sample cuvette.
 Photosensitive Detectors
 The detectors are devices that convert radiant energy into electrical signal.
 A Detector should be sensitive, and has a fast response over a considerable
range of wavelengths.

 In addition, the electrical signal produced by the detector must be directly

proportionate to the transmitted intensity.
Amplifier
•The alternating current generated in the photocells is transferred to the amplifier.
•The amplifier is coupled to a small servometer.
•Current generated in the photocells is of very low intensity, amplifier, amplify the
signals many times so we can get clear and recordable signals.
Recording devices
•Most of the time amplifier is coupled to a pen recorder which is connected to the
computer.
•Computer stores all the data generated and produces the spectrum of the desired
compound.
 Types
The components of a single beam spectrophotometer
Light source - white light of constant intensity

slits

Grating
slits Separates white light
Phototube
into various colors
detects light &
measures intensity Rotating the grating
Sample
changes the wavelength
When blank is the sample going through the sample
Po is determined,
otherwise P is measured
 Double Beam Spectrophotometer

Beam Chopper Semi-transparent

Mirror
Sample

Tungsten Slit Quartz Photo-

Grating
Lamp Cuvette multiplier
Reference
(Blank)
Mirror Mirror
III. Chromophores: Structural feature causing absorption of UV/Vis light
C. Substituent Effects
i. Bathochromic shift (red shift) – a shift to longer wavelength; 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

Hyperchromic

Hypsochromic Bathochromic

Hypochromic

200 nm 700 nm
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IV. Structure Determination
C. Aromatic Compounds
1. The allowed transition is not in the routine range of UV obs. at 180 nm, and is
referred to as the primary band
2. The forbidden transition is observed if substituent effects shift it into the obs. Region
204 nm; this is referred to as the second primary band
3. At 260 nm is another forbidden transition referred to as the secondary band.

Compound Peak 1 (in nm/ε) Peak 2 (in nm//ε) Peak 3 (in nm/ε) Color
Benzene 184/60000 204/7900 256/200 colorless liquid
Naphthalene 221/133000 286/9300 312/289 white solid
Anthracene 256/180000 375/9000 pale yellow
Naphthacene 278/170000 474/10000 yellow

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 Structural characterizations
transition with
Compound λ(nm) Intensity/ε
lowest energy
CH4 122 intense σ-σ*(C-H)
CH3CH3 130 intense σ-σ* (C-C)
CH3OH 183 200 n-σ* (C-O)
CH3SH 235 180 n-σ* (C-S)
CH3NH2 210 800 n-σ* (C-N)
CH3Cl 173 200 n-σ* (C-Cl)
CH3I 258 380 n-σ* (C-I)
CH2=CH2 165 16000 π-π* (C=C)
CH3COCH3 187 950 π-π* (C=O)
273 14 n-π* (C=O)
CH3CSCH3 460 weak n-π* (C=S)
CH3N=NCH3 347 15 n-π* (N=N)
The portion of the EM spectrum from 400-800 is observable to humans- we (and some
other mammals) have the adaptation of seeing color at the expense of greater detail

400 500 600 700 800

l, nm

Violet 400-420
Indigo 420-440
Blue 440-490
Green 490-570
Yellow 570-585
Orange 585-620
Red 620-780
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When white (continuum of l) light passes through, or is reflected by a surface, those
ls that are absorbed l are removed from the transmitted or reflected light respectively
What is “seen” is the complimentary colors (those that are not absorbed)
This is the origin of the “color wheel”

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FTIR
 IR radiations

The wavelength of IR radiation is 800 nm – 1 mm (106 nm).

Energy 1-15 kcal/mole.

The frequency (Hz) of infrared radiations is 430 THz -300 GHz.

E of IR is 1.7 Ev-1.24 MeV.

 Very Important
Near IR 12,500 to 4,000 cm-1 800 nm to 2.5 μm
Mid IR 4000-400 cm-1 (2.5-25 μm)
Far-IR 400-10 cm-1 (25-1000 μm),

Functional group region 4000 cm-1 to 1450 cm-1

Fingerprint region from 1450 cm-1 to 400 cm-1.

FTIR gives data Time vs Intensity which is then fourier transformed to wavenumber vs
Intensity spectra.

Dispersive IR gives Frequency vs Intensity data which is then converted to

wavenumber vs Intensity spectra.
 Infrared active and inactive bonds
 Not all covalent bonds display bands in the IR, only polar bonds do so. These are referred
to as IR active. (N2 and O2 in air are IR inactive)
 Strongly polar bonds such as carbonyl groups (C=O) produce strong bands.
 Medium polarity bonds and asymmetric bonds produce medium bands.
 Weakly polar bond and symmetric bonds produce weak or non-observable bands.
 Vibrational modes

H H
C C
H H
Symmetric stretching Asymmetric Stretching (higher E)

H H H H

C C C C
H H
H H
wagging
Scissoring Rocking twisting

In plane bending out of plane

 Dispersive IR
1. There are many moving parts resulting in
mechanical slippage and wear.
2. Calibration against reference spectra is required to
measure frequency.
3. Slow scanning speed.
4. In order to improve resolution only a small amount
of the IR beam may be allowed to pass through slits.
5. Only radiation of a narrow frequency range falls on
the detector at any one time .
6. The samples is subject to thermal effect from the
focused beam .
7. Less sensitive, time consuming.
Fourier Transform IR
Only the mirror moves during an experiment
Use of a laser provides high frequency (to 0.01 cm-1).
Rapid scan speeds permit monitoring sample
undergoing rapid change.
A much larger beam may be used at all times. Data
collection is easier.
All frequencies of radiation fall on the detector
simultaneously .
The sample is not subject to thermal effects.
High sensitivity, precision & faster
 Sample Preparation
Solid sample
Solids run in solution

Solid is dissolved in a non-aqueous solvent (non reactive and is not absorbed IR).

A drop is placed on the alkali metal disc, solvent is evaporated leaving a thin film of the solute.

Case film technique

Amorphous solid is deposited from the melt on the surface of a KBr or NaCl cell

Ensured that the film is not too thick to pass the radiation.

Solvents

dry and transparent in the region of interest. A common pair of solvents is CCl4 and CS2 .
 Solid sample

Mull technique

• Crushed sample (2-5mg) is mixed with Nujol (mulling agent/HC) to make

thick paste.

• A thin film is applied onto the salt plates.

• This is then mounted in a path of IR beam and the spectrum is recorded.

 Solid sample preparation
Pressed pellet technique

Small amount of finely ground solid sample is mixed with 100 times its weight of KBr
Compressed into a thin transparent pellet using a hydraulic press.
These pellets are transparent to IR radiation and it is used for analysis.
 Liquid sample

A drop of a liquid organic compound is placed between a pair of polished

sodium chloride or potassium bromide plates, referred to as salt plates.

Dilution with appropriate solvent may be necessary (CCl4, CS2, CH3Cl).

Solvent must be transparent in the region of interest.

Salt plates are hygroscopic and water soluble (avoid water).

Special cells are used for water containing samples (BaF2, AgO).
 Gas sample
Gas samples are enclosed in a gas cell with windows,

The length of the cell can be centimeters to over 10 meters.

This cell is placed in the beam path much like the solid or liquid sample with the
exception that gas cell has two windows that the IR beam needs to pass through.

Temperature and pressure are controlled if quantitative analysis is required.

 Radiation sources

The radiation source is composed of an inert solid which is electrically heated to a

temperature in the range 1500-2000 K. The heated material will then emit infrared
radiation.

Nernst glower

 Globar source

 Incandescent lamp

 Tungsten lamp
 Radiation source
Nernst Glower
• Composed of rare earth oxides e.g. zirconium oxide, yttrium oxide.
• Platinum ends leads to passage of electricity.
• Temperature: 1000-1800 oC.

 Used as thermal light source.

It is a silicon carbide rod of 5 to 10 mm
diameter and 20 to 50 mm length.
It is electrically heated up to 1000 to 1650 °C.
Incandescent lamp
Temperature can be raised to 1100 oC.
 Nichrome wire is tightly wound.
 Gives less energy long life then Nernst and Goblar.
 Michelson interferometer
core of FTIR spectrometers, splits one beam into two
so that the paths of the two beams are different.

Then the Michelson interferometer recombines the

two beams and conducts them into the detector where
the difference of the intensity of these two beams are
measured as a function of the difference of the paths.
 Detectors
(1). Thermal detector:
Thermocouple
Thermistors (
Pyroelectric (
Bolometer (
(2). Photodetector:
Photodiodes:

Photomultiplier Tube:
 Effect of H-bonding
In UV/Vis: When hydrogen bonds were formed between these molecules, a blue shift in the
absorption maximum is observed in addition to a decrease in the total absorption intensity.

IN FTIR: The shift in frequency of about 300cm−1 arises because H-bonding weakens the
O−H bond; its absorption frequency then will be lower.
The association band is broad.

What is the effect of H-bonding on chemical shift in NMR?

Thank You