Spectroscopy

• Four common spectroscopic techniques used

to determine structure:

– Infrared Spectroscopy (IR)

– Nuclear Magnetic Resonance Spectroscopy (NMR)
– Ultraviolet Spectroscopy
– Mass Spectrometry (MS or Mass Spec)
2
electromagnetic radiation
The position of a band is reported in wavenumbers ( u )
the number of wavelengths per cm

E = h = hc/l

Directly proportional
to energy
 Spectroscopy
The spectra
Molar Absorptivity (l/mol-cm) Electronic Spectra

51972

41578

31183

20789

10394

Nacetylindol

0 Wavelength (nm)

Infrared spectroscopy (IR)

220 230 240 250 260 270 280 290 300

(Ultraviolet / VIS) - reports on functional groups

conjugation and multiple bonds.

Nuclear magnetic resonance (NMR) Mass spectrometry (MS)*

C/H molecular skeleton molecular formula
 INTRODUCTION
IR spectroscopy is Absorption spectroscopy in which molecular
vibrations observed due to absorption of IR radiation.

Infrared radiation was discovered in

1800 by William Herschel.

5
Infrared spectroscopy is a type of absorption
spectroscopy:
– Where the amount of light absorbed by (or transmitted
through) a compound is measured as a function of the
wavelength of light
• Sample is irradiated by a light source

• Amount of light transmitted (or absorbed) at various

wavelengths is measured by a detector

• A spectrum is obtained.
– Graph of light transmitted (or absorbed) as a
function of wavelength
 Pharmaceutical Applications

 Qualitative determination of substances

 Structural elucidation by determining the functional
groups.
 Detection of impurities
 Identification of geometrical isomers for both organic
& inorganic samples
 Detection of presence of water in sample

7
 Infrared Spectroscopy
• The structure of new compounds that are
isolated from natural sources or prepared in
the lab must be determined (and/or verified).
– Chemical analysis
– Spectroscopy

• Spectroscopic techniques are non-destructive

and generally require small amounts of
sample
 Infrared Spectroscopy
Infrared Spectroscopy
• The atoms in a molecule are in constant motion.
• The covalent bond between two atoms acts like a
spring, allowing the atoms to vibrate (stretch and
bend) relative to each other.

Bonded atoms having different strengths due to different masses.

Absorption of IR energy will cause vibrational changes in molecule and a peak will be
observed, when
Conditions to obtain IR spectrum
1) Dipole moment of bonds changes during vibration.
- Vibrations that cause a significant change in the dipole
moment of a chemical bond lead to strong absorption
bands.
- Vibrations that result in no change/very little change in
dipole moment lead to very weak or no absorption
band.

• Symmetrical bonds often exhibit very weak or no

absorption band.

1) When covalent bonds are polar in nature

When sample obeys these conditions, then it gets vibrated

by absorbing radiation and gives IR spectrum.
10
 TYPES OF VIBRATIONS

Stretching Mode

Bending vibrations:

•These are also called as

deformations.
•In this bond angle is
altered.

•These are of 2 types

a) in plane bending→
scissoring, rocking
Symmetric Asymmetric
b) out plane bending→
wagging, twisting 11
 TYPES OF VIBRATIONS

In plane bending vibrations

It is an out of plane bending.
In this 2 atoms move to one
side of the plane. They move
up and down the plane.

Scissoring Rocking

Out plane bending vibrations

Twisting:
•In this one atom moves above
the plane and the other atom
moves below the plane.
Wagging Twisting 12
 Infrared Spectroscopy
• The frequency of an absorption band in an IR
spectrum depends primarily on:

– Type of vibration
• Stretching vibrations: higher frequency
• Bending vibrations: lower frequency

– Masses of the atoms in a bond

AW Freq
– Strength of the bond or bond order
BO Freq
 Factors affecting Infrared Absorptions
• Symmetry- has no dipole moment, therefore does not generate
oscillating dipole.
(vibrations give rise to absorption if it is associated with oscillating electric dipole)

• Coupling of vibrations
– in-phase and out-of phase
– Stretching and bending

• Hydrogen bonding
– Broadening of peaks

• Ring strain
– Frequency increases with increase in angle strains

• Electronic Effects
– Resonance and inductive effects (higher freq with inductive effect eg acid
chloride= 1800 while aldehyde=1730cm-1)

• Isomerism
– Constitutional, stereoisomers, conformational and tautomerism
 Radiation sources
They must produce intense & steady radiation.

Nernst Glower heated rare earth oxide rod 1-50 µm

(zirconium, yttria, thoria) (~1500 (mid- to far-IR)
K)
Globar heated Silicon Carbide rod 1-50 µm
(~1500 K) (mid- to far-IR)
W filament lamp 1100 K 0.78-2.5 µm
(Near-IR)
Hg arc lamp Hg plasma 50 - 300 µm
(far-IR)
15
 Monochromators
These allow for filtration of desired frequency of radiation
Monochromators are 2 types
1) Prismatic Monochromators:
Composed of glass or quartz and coated by alkyl halides (NaCl)

1) Grating Monochromators:
High dispersion and resolution than prismatic monochromators
Reflection Grating Monochromator are common than
Transmittance Gratings

16
 Detectors/Transducers

Thermocouple Thermoelectric effect - cheap, slow,

dissimilar metal junction insensitive

Bolometer Ni, Pt resistance Highly sensitive

thermometer (thermistor) <400 cm-1

Golay cell Metal cylinder with Xe gas
Pyro electric Tri glycine sulfate
piezoelectric material
Faster than others &
having wide
wavelength range
fast and sensitive
(mid IR)

Photoconductive PbS, CdS, Pb Se light sensitive fast and sensitive

(non-thermal) cells (near IR) 17
 Dispersive IR instrument
Sequential scanning of each wave number takes place
Introduced in 1940’s.

• In double beam spectrometer , beam separates into two and

passes to sample & reference.
• Prismatic monochromators have been replaced with Grating
monochromator.
• In dispersive IR sequential scanning of wave numbers of
light takes place
• Dispersive IR failed due to monochromator containing
narrow slits which limit the wave number of radiation.
18
 FTIR Instrumentation
FTIR collects all wavelengths simultaneously and scans at once.
Beam splitter
Fixed mirror
Movable mirror

Interferometer

Widely applied and quite popular in the far-IR

and mid-IR spectrometry. 19
 Advantages
 Fast & sensitive
 All frequencies can be modulated at once
 Simple mechanical design with only one moving part
 No stray light is involved
 Availability of easy sampling accessories
 Air pollutants like CO, ethylene oxide etc. can be analysed

20
 Sample Preparation
• Solids- MULL

– Small amount of sample is ground in agate or mullite mortar with

mulling material (colourless paraffin oil) in a thin paste.

– Sample is then transferred to a plate of NaCl or KBr and covered

with another plate.

– A thin film is produced by gentle pressure with a slight rotation.

– 2 plates with mull is placed in cell holder and the spectrum

recorded.

– Strong bands at 2900cm-1, 1470cm-1and 1370 cm-1 and weak band

at 720 cm-1is due to paraffin.
 Sample Preparation
• Solids- PRESSED PELLET

– A few milligrams of sample is ground with 100x the

quantity of matrix material (transparent material: KBr used
for this)

– Ground powder is transferred to stainless steel dice

(~13mm in diameter).

– Power is pressed into a disc between stainless steel dice.

– Well made pellet will have 80-90% transmittance in regions

below 3000cm-1.
 Sample Preparation

• Liquid and solutions

– Place one drop of the liquid onto a disc of NaCl or

KBr.

– Cover the drop with another disc.

– Mount the discs on the holder and run the spectra.

– Teflon spacers may be used to give various

pathlengths.
 Infrared Spectroscopy
• Each molecule has a unique IR spectrum.
– The IR spectrum is a “fingerprint” for the molecule.

• IR spectrum results from a combination of all possible

stretching and/or bending vibrations of the individual bonds
and the whole molecule.
– Simple stretching: ~1500-4000 cm-1.
– Complex vibrations: 600-1400 cm-1, called the
“fingerprint region.”
 Regions of IR Spectra
• 4000-2500 cm-1 N-H, C-H, O-H • 2000-1500 cm-1 double bonds
(stretching) (stretching)
• 3300-3600 cm-1 N-H, O-H •C=O 1680-1750 cm-1
• 3000 cm-1 C-H •C=C 1640-1680 cm-1
• 2500-2000 cm-1 C≡C and C≡N • Below 1500 cm-1 “fingerprint”
(stretching) region
 IR Spectroscopy
Example: IR spectrum of the some major peaks. Identify
functional group(s) present.

3403
cm-1

1604
cm-1
 O-H and N-H Bonds
• Both O-H and N-H stretches appear around
3300 cm-1, but they look different.
– Alcohol O-H
• broad with rounded tip when hydrogen bonding is
present (sharp in the absence of hydrogen bonding)
– Secondary amine (R2NH)
• Broad (usually) with one sharp spike
– Primary amine (RNH2)
• Broad (usually) with two sharp spikes.
– No signal for a tertiary amine (R3N)
 Infrared Spectroscopy
• OH

http://riodb01.ibase.aist.go.jp/sdbs/ (National Institute of Advanced Industrial Science and Technology,

1/5/11)
 NH Bend
• A broad, round peak may be observed around 1600 cm-1 for the
N – H bend, especially with primary amines.

NH2
stretch
N-H
bend
N-H bend has
a different
shape than an
aromatic ring
or C=C
 Carbon-Hydrogen Bonds

• Bonds with more s character absorb at a higher

frequency.
– sp3 (alkane) C-H
• just below 3000 cm-1 (to the right)

– sp2 (alkene or aromatic hydrocarbon) C-H

• just above 3000 cm-1 (to the left)

– sp (alkyne) C-H
• at 3300 cm-1
 Infrared Spectroscopy
• IR Spectrum of n-octane
 Carbon-Carbon Bonds
• Increasing bond order leads to higher frequencies:
– C-C 1200 cm-1 (fingerprint region)
– C=C 1600 - 1680 cm-1
– CC 2200 cm-1 (weak or absent if internal)

• Conjugation lowers the frequency:

– isolated C=C 1640-1680 cm-1
– conjugated C=C 1620-1640 cm-1
– aromatic C=C approx. 1600 cm-1

• C=C peaks are generally weak to moderate in intensity.

 Carbonyls
• Carbonyl stretches are generally strong:

– Aldehyde ~1710 cm-1

– Ketone ~1710 cm-1
– Carboxylic acid ~1710 cm-1
– Ester ~1730 - 1740 cm-1
– Amide ~1640-1680 cm-1

• Conjugation shifts all carbonyls to lower frequencies.

• Ring strain shifts carbonyls to higher frequencies. -1

1745 cm

O
H3C
Aldehydes
Carboxylic Acids
Ketones
 Esters
– C=O stretch at ~ 1730-1740 cm-1
strong
and
– C-O stretch at 1000-1300 cm-1 (broad)
(Note: other functional groups may have peaks in the 1000-1300 cm-1
region too!)
O

1743 1245
 Amides
• C=O stretch at 1640-1680 cm-1 (sometimes a double
peak)
• N-H stretch (if 1o or 2o) around 3300 cm-1
 Nitriles
• C  N absorbs just above 2200 cm-1 (med – strong)

• The alkyne C  C signal is much weaker and is just below

2200 cm-1
 IR Spectroscopy
Example: Interpret the following IR spectrum by assigning
each of the major peaks. Identify what functional group(s)
are present.

3403
cm-1

1604
cm-1

http://riodb01.ibase.aist.go.jp/sdbs/ (National Institute of Advanced Industrial Science and Technology,

12/30/09)
 IR Spectroscopy
Example: Interpret the following IR spectrum by
assigning each of the major peaks. Identify what
functional group(s) are present.

2733
cm-1
2814
cm-1

1691 1642
cm-1 cm-1

http://riodb01.ibase.aist.go.jp/sdbs/ (National Institute of Advanced Industrial Science and Technology,

12/30/09)
 Infrared Spectroscopy
Example: Which one of the following compounds is the
most reasonable structure for the IR spectrum shown
below? O O
OCH 3 OH
O O

1721

http://riodb01.ibase.aist.go.jp/sdbs/ (National Institute of Advanced Industrial Science and Technology,

12/30/09)
 Infrared Spectroscopy
Example: Which of the following vibrations will show in
the IR spectrum shown of CO2?
 Infrared Spectroscopy
Example: Which of the following vibrations will show in
the IR spectrum shown of CO2?
Identify the structure