University of Al-Ameed

College of Pharmacy

Advanced pharmaceutical analysis

lecture 3
5th class/ 2nd semester

Infrared spectroscopy
IR Spectroscopy

I. Introduction

A. Spectroscopy is the study of the interaction of matter with the

electromagnetic spectrum

1. Electromagnetic radiation displays the properties of both particles and

waves

2. The particle component is called a photon

3. The energy (E) component of a photon is proportional to the frequency.

Where h is Planck’s constant and n is the frequency in Hertz (cycles per
second)

E = hn

4. The term ―photon‖ is implied to mean a small, massless particle that

contains a small wave-packet of EM radiation/light – we will use this
terminology in the course
2
IR Spectroscopy

5. Because the speed of light, c, is constant, the frequency, n, (number

of cycles of the wave per second) can complete in the same time, must
be inversely proportional to how long the oscillation is, or wavelength:
c
___ hc
n= ___
 E = hn =
l l
c = 3 x 1010 cm/s

6. Amplitude, A, describes the wave height, or strength of the oscillation

7. Because the atomic particles in matter also exhibit wave and particle
properties (though opposite in how much) EM radiation can interact
with matter in two ways:
• Collision – particle-to-particle – energy is lost as heat and
movement

• Coupling – the wave property of the radiation matches the wave

property of the particle and ―couple‖ to the next higher quantum
mechanical energy level
3
IR Spectroscopy

8. The entire electromagnetic spectrum:

Frequency, n in Hz
~1019 ~1017 ~1015 ~1013 ~1010 ~105

Wavelength, l
~.0001 nm ~0.01 nm 10 nm 1000 nm 0.01 cm 100 m

Energy (kcal/mol)
> 300 300-30 300-30 ~10-4 ~10-6

g-rays X-rays UV IR Microwave Radio

nuclear core electronic molecular molecular Nuclear Magnetic
excitation electron excitation vibration rotation Resonance NMR
(PET) excitation (p to p*) (MRI)
(X-ray
cryst.)

Visible
4
IR Spectroscopy

The Electromagnetic Spectrum

5
IR Spectroscopy

I. Introduction

B. The IR Spectroscopic Process:

1. The quantum mechanical energy levels observed in IR spectroscopy are
those of molecular vibrations.

2. We perceive this vibration as heat.

3. When we say a covalent bond between two atoms is of a certain

length, we are citing an average because the bond behaves as if it were
a vibrating spring connecting the two atoms.

4. For a simple diatomic molecule, this model is easy to visualize:

6
IR Spectroscopy

5. There are two types of bond vibration:

• Stretch – Vibration or oscillation along the line of the bond

H H
C C
H H
symmetric asymmetric

• Bend – Vibration or oscillation not along the line of the bond

in plane out of plane

H H H H
C C C C
H H H H

scissor rock twist wag

7
IR Spectroscopy

Vibrational Modes for a CH2 Group

8
IR Spectroscopy

6. As a covalent bond oscillates – due to the oscillation of the dipole

of the molecule – a varying electromagnetic field is produced

7. The greater the dipole moment change through the vibration, the
more intense the EM field that is generated.

9
 Hooke’s law
• Bonds can be thought like a spring, and
wavenumbers can be approximated by Hooke’s
law
• Hooke’s law calculate the relationship
between wavenumber of oscillation, atomic
masses, and bond force constant.

• single bond 5 x105 dyne/cm

• double bond 10 x105 dyne/cm
• triple bond 15 x105 dyne/cm

The calculation is not highly accurate because effects arising from the
molecule environment.
Hooke’s law-Example
12
IR Spectroscopy

Regions of transparency for common infrared solvents.

The horizontal lines indicate regions where solvent transmits at least 25% of
the incident radiation in a 1-mm cell.

13
IR Spectroscopy

C. The IR Spectrum
1. Each stretching and bending vibration occurs with a characteristic
frequency as the atoms and charges involved are different for different
bonds
The y-axis on an IR In regions where
spectrum is in units of no osc. bond is
% transmittance interacting with
IR light,
transmittance
In regions where the EM nears 100%
field of an osc. bond
interacts with IR light of
the same n –
transmittance is low
hence IR light is
absorbed.

14
IR Spectroscopy

C. The IR Spectrum

2. The x-axis of the IR spectrum is in units of wavenumbers, n, which is the

number of waves per centimeter in units of cm-1 (Remember E = hn or E
= hc/l)

15
IR Spectroscopy

C. The IR Spectrum

3. This unit is used rather than wavelength (microns) because

wavenumbers are directly proportional to the energy of transition
being observed.
High frequencies and high wavenumbers equate higher energy
is quicker to understand than Short wavelengths equate higher
energy.

4. This unit is used rather than frequency as the numbers are more
―real‖ than the exponential units of frequency

5. IR spectra are observed for the mid-infrared: wavelengths: 2.5 - 50

m, wavenumbers: 4000 - 200 cm-1

6. The peaks are Gaussian distributions of the average energy of a

transition

16
IR Spectroscopy

C. The IR Spectrum

7. In general:
Lighter atoms will allow the oscillation to be faster – higher energy
This is especially true of bonds to hydrogen – C-H, N-H and O-H
Stronger bonds will have higher energy oscillations
Triple bonds > double bonds > single bonds in energy

Energy/n of oscillation
IR Spectroscopy

C. The IR Spectrum -The detection of different bonds

7. As opposed to chromatography or other spectroscopic methods, the area of

a IR band (or peak) is not directly proportional to concentration of the
functional group producing the peak

8. The intensity of an IR band is affected by two primary factors:

 Whether the vibration is one of stretching or bending

 Electronegativity difference of the atoms involved in the bond

• For both effects, the greater the change in dipole moment in a given
vibration or bend, the larger the peak.

• The greater the difference in electronegativity between the atoms

involved in bonding, the larger the dipole moment

• Typically, stretching will change dipole moment more than bending

18
IR Spectroscopy

C. The IR Spectrum – The detection of different bonds

9. It is important to make note of peak intensities to show the effect of

these factors:

• Strong (s) – peak is tall, transmittance is low (0-35 %)

• Medium (m) – peak is mid-height (75-35%)

• Weak (w) – peak is short, transmittance is high (90-75%)

• * Broad (br) – if the Gaussian distribution is abnormally broad

(*this is more for describing a bond that spans many energies)

Exact transmittance values are rarely recorded

19
Infrared Spectroscopy

II. Infrared Group Analysis

A. General
1. The primary use of the IR is to detect functional groups.

2. Because the IR looks at the interaction of the EM spectrum

with actual bonds, it provides a unique qualitative probe into
the functionality of a molecule, as functional groups are merely
different configurations of different types of bonds.

3. Since most “types” of bonds in covalent molecules have

roughly the same energy, i.e., C=C and C=O bonds, C-H and
N-H bonds they show up in similar regions of the IR spectrum.

4. Remember all organic functional groups are made of multiple

bonds and therefore show up as multiple IR bands (peaks)
Infrared Spectroscopy

II. Infrared Group Analysis

A. General
5. The four primary regions of the IR spectrum

Bonds to H Triple bonds Double bonds Single Bonds

Fingerprint
Region

O-H C≡C C=O C-C

N-H C≡N C=N C-N
C-H C=C C-O

4000 cm-1 2700 cm-1 2000 cm-1 1600 cm-1 600 cm-1
Infrared Spectroscopy
Octane
1. Alkanes – combination of C-C and C-H bonds
• C-C stretches and bends 1360-1470 cm-1

• CH2-CH2 bond 1450-1470 cm-1

• CH2-CH3 bond 1360-1390 cm-1

• C-H of (CH3 ) between 2800-3000 cm-1

• (w= weak, m= moderate, and s= strong peaks)

(w – s) (m)
Infrared Spectroscopy
1-Octene
2. Alkenes – addition of the C=C and vinyl C-H bonds
• C=C stretch at 1620-1680 cm-1 weaker as
substitution increases

• vinyl C-H stretch occurs at 3000-3100 cm-1

• The difference between alkane, alkene or

alkyne C-H is important:
• If the band is slightly above 3000 it is alkene or
alkyne C-H.
• If it is below 3000 cm-1 it is alkane C-H.

(w – m)
(w – m)
Infrared Spectroscopy
1-Octyne
3. Alkynes – addition of the C≡C and alkyne terminal C-H
bonds:
• C≡C stretch 2100-2260 cm-1; strength depends on
asymmetry of bond, strongest for terminal alkynes,
weakest for symmetrical internal alkynes.

• C-H for terminal alkynes occurs at 3200-3300cm-1

• Internal alkynes ( R-C≡C-R ) would not have this band!

(w-m)

(m – s)
Infrared Spectroscopy
Ethyl benzene
4. Aromatics
• Due to the delocalization of e- in the ring, the stretching
frequency for these bonds is slightly lower in energy
than normal C=C.
• These show up as a pair of sharp bands, 1500 & 1600
cm-1.

• C-H bonds off the ring show up similar to vinyl C-H at

3000-3100 cm-1

Overtone region

(w – m) (w – m)
Infrared Spectroscopy
4. Aromatics
• If the region between 1667-2000 cm-1 (w) is free of interference (C=O stretching
frequency is in this region) a weak grouping of peaks is observed for aromatic
systems
• Analysis of this region, called the overtone of bending region, can lead to a
determination of the substitution pattern on the aromatic ring.

Monosubstituted
G
G
1,2 disubstituted (ortho or o-)

G
1,2 disubstituted (meta or m-)
G
G
1,4 disubstituted (para or p-)

G
Infrared Spectroscopy

5. Unsaturated Systems – substitution patterns

• The substitution of aromatics and alkenes can also be discerned through the out-
of-plane bending vibration region
• However, other peaks often are apparent in this region. These peaks should only
be used for reinforcement of what is known or for hypothesizing as to the
functional pattern.
cm-1 cm-1
R
985-997 730-770
C CH2 905-915 R
690-710
H

R H
C C 960-980 R 735-770
H R
R
R R
C C 665-730 860-900
H H R 750-810
680-725
R
R
C CH2 885-895
R
R R 800-860
R R
C C 790-840
R H
Infrared Spectroscopy
Diisopropyl ether
6. Ethers –
• Show a strong band for the symmetric C-O-C
stretch at 1050-1150 cm-1

(s)
Infrared Spectroscopy
1-butanol
7. Alcohols
• Strong, broad O-H stretch from 3200-3400 cm-1

• Like ethers, C-O stretch from 1050-1260 cm-1

• Band position changes depending on the alcohols

substitution: 1° 1000-1075; 2° 1075-1150; 3° 1100-1200;
phenol 1180-1260

• The shape is due to the presence of hydrogen bonding

(m– s)
br

(s)
Infrared Spectroscopy
2-aminopentane
8. Amines - Primary
• Shows the –N-H stretch for NH2 as a doublet
between 3200-3500 cm-1 (symmetric and
asymmetric (anti-symmetric) modes)
• -NH2 has deformation band from 1590-1650 cm-1
• Additionally there is a “wag” band at 780-820 cm-1
that is not diagnostic

(w) (w)
Infrared Spectroscopy
pyrrolidine
9. Amines – Secondary
• N-H band for R2N-H occurs at 3200-3500 cm-1
as a single sharp peak weaker than –O-H

• Tertiary amines (R3N) have no N-H bond and

will not have a band in this region

(w – m)
Infrared Spectroscopy

Pause and Review

• Inspect the bonds to H region (2700 – 4000 cm-1)

• Peaks from 2850-3000 are simply sp3 C-H in most organic molecules

V-shape peak
-N—H bond for 2o
Broad U-shape peak amine (R2N—H)
-O—H bond
W-shape peak
-N—H bond for 1o
amine (RNH2)
Sharp spike 3000 cm-1
-C≡C—H bond
Small peak shouldered
just above 3000 cm-1
C=C—H or Ph—H
Infrared Spectroscopy

10. Aldehydes
• C=O (carbonyl) stretch from 1720-1740 cm-1
• Band is sensitive to conjugation, as are all carbonyls (upcoming slide)
• A highly unique sp2 C-H stretch appears as a doublet, 2720 & 2820 cm-1 called a “Fermi doublet”

(w-m)

(s)
Infrared Spectroscopy
3-methyl-2-pentanone
11. Ketones
• Simplest of the carbonyl compounds as far as IR
spectrum – carbonyl only

• C=O stretch occurs at 1705-1725 cm-1

(s)
Infrared Spectroscopy
Ethyl pivalate
12. Esters
• C=O stretch at 1735-1750 cm-1

• Strong band for C-O at a higher frequency than

ethers or alcohols at 1150-1250 cm-1

(s)
(s)
Infrared Spectroscopy
4-phenylbutyric acid
13. Carboxylic Acids:
• Gives the messiest of IR spectra

• C=O band occurs between 1700-1725 cm-1

• The highly dissociated O-H bond has a broad band

from 2400-3500 cm-1 covering up to half the IR
spectrum in some cases

(w – m) (s) (s)
br
Infrared Spectroscopy
Propionic anhydride
14. Acid anhydrides O O

• Coupling of the anhydride though the ether oxygen

splits the carbonyl band into two with a separation O
of 70 cm-1

• Bands are at 1740-1770 cm-1 and 1810-1840 cm-1

• Mixed mode C-O stretch at 1000-1100 cm-1

(s) (s)
Infrared Spectroscopy
Propionyl chloride
O
15. Acid halides
• Clefted band at 1770-1820 cm-1 for C=O
Cl
• Bonds to halogens, due to their size occur at low
frequencies, only Cl is light enough to have a band
on IR, C-Cl is at 600-800 cm-1

(s)

(s)
Infrared Spectroscopy
pivalamide
O
16. Amides
• Display features of amines and carbonyl compounds
• C=O stretch at 1640-1680 cm-1 NH2

• If the amide is primary (-NH2) the N-H stretch occurs from

3200-3500 cm-1 as a doublet
• If the amide is secondary (-NHR) the N-H stretch occurs at
3200-3500 cm-1 as a sharp singlet

(m – s) (s)
Infrared Spectroscopy

17. Nitro group (-NO2)

2-nitropropane
• Proper Lewis structure gives a bond order of 1.5 from
nitrogen to each oxygen O
O
N
• Two bands are seen (symmetric and asymmetric) at
1300-1380 cm-1 and 1500-1570 cm-1

• This group is a strong resonance withdrawing group and

is itself vulnerable (susceptible) to resonance effects

(s) (s)
Infrared Spectroscopy

18. Nitriles (the cyano- or –C≡N group) propionitrile

• Principle group is the carbon nitrogen triple bond at 2100-2280 cm-1 N
• This peak is usually much more intense than that of the alkyne due to C
the electronegativity difference between carbon and nitrogen

(s)
Infrared Spectroscopy
Fingerprint region
In the region from  1300 to 400 cm-1, vibrational frequencies are affected by the
entire molecule, as the broader ranges for group absorptions in the figure below –
fingerprint region.
Absorption in this fingerprint region is characteristic of the molecule as a whole. This
region finds widespread use for identification purpose by comparison with library
spectra.
H.W: does the fingerprint region useful in characterization of drug molecules?
43
Infrared Spectroscopy

Effects on IR bands
1. Conjugation – by resonance, conjugation lowers the energy of a double or triple
bond. The effect of this is readily observed in the IR spectrum:
O
O

1684 cm -1 1715 cm -1
C=O C=O
• Conjugation will lower the observed IR band for a carbonyl from 20-40 cm-1
provided conjugation gives a strong resonance contributor
O
C X X= NH 2 CH 3 Cl NO 2
H3 C
1677 1687 1692 1700 cm-1

O O
O
N C
H 2N C CH 3 vs.
CH3
O

Stron g resonance con tributor Poor reso nance contributo r

(canno t resonate with C=O)
• Inductive effects are usually small, unless coupled with a resonance
contributor (note –CH3 and –Cl above)
Infrared Spectroscopy

Effects on IR bands
2. Steric effects – usually not important in IR spectroscopy, unless they reduce the
strength of a bond (usually p) by interfering with proper orbital overlap:
O O

CH 3
C=O : 1686 cm-1 C=O : 1693 cm-1
• Here the methyl group in the structure at the right causes the carbonyl
group to be slightly out of plane, interfering with resonance

3. Strain effects – changes in bond angle forced by the constraints of a ring will
cause a slight change in hybridization, and therefore, bond strength

O O O
O O

1815 cm-1 1775 cm-1 1750 cm-1 1715 cm-1 1705 cm-1

• As bond angle decreases, carbon becomes more electronegative, as well

as less sp2 hybridized (bond angle < 120°)
Infrared Spectroscopy

Effects on IR bands
4. Hydrogen bonding
• Hydrogen bonding causes a broadening in the band due to the creation
of a continuum of bond energies associated with it
 In the solution phase these effects are readily apparent;
 in the gas phase where these effects disappear or in lieu of steric effects,
the band appears as sharp as all other IR bands:

Gas phase spectrum of

1-butanol

Steric hindrance to H-bonding

in a di-tert-butylphenol
OH

• H-bonding can interact with other functional groups to lower frequencies

H
O O

C=O; 1701 cm-1

Intermolecular H-bonding increases as the
concentration of the solution increases or when the
sample is analysed neat. This lowers frequency of OH
absorption to 3550-3200cm-1.

This is because the stronger the hydrogen-bonding

causes:
• the longer the O-H bond (more polarized)
• the lower the force constant
• the lower the vibrational frequency
• the broader the band (H-bonding is not uniform
throughout the sample) and the more intense (due to
increased polarization of the O-H bond) the absorption
band.
48
5- Electronic Effect of Substituents (affecting position of C=O
absorption)
Replacement of an alkyl group of a saturated aliphatic
ketone by heteroatom (X) shifts the carbonyl absorption.
The direction of the shift depends on whether inductive
effect or resonance effect predominates.

Inductive effect reduces the length of the C=O bond,

therefore, increases its force constant and the frequency
of absorption will increase.

49
Resonance effect increases the length of the C=O
bond, therefore, reduces its force constant and the
frequency of absorption will decrease.

50
(for monomer)

51
Can IR spectroscopy distinguish the following
isomers?

52
Infrared Spectroscopy
Main uses of IR spectroscopy:
1. Fundamental chemistry:
a. Determination of molecular structure.
b. Determination of molecular geometry.
e.g. Determination of bond lengths, bond angles of gaseous molecules.
1. Qualitative analysis: – simple, fast, nondestructive
i. Monitoring trace gases: NDIR.
ii. Rapid, simultaneous analysis of GC, moisture, N in soil.
iii. Analysis of fragments left at the scene (place) of a crime.
2. Quantitative analysis:
A. determination of hydrocarbons on filters.
B. determination of hydrocarbons in air.
C. determination of hydrocarbons in water
Infrared Spectroscopy

Near-infrared and Far-infrared absorption

The techniques and applications of near-infrared (NIR) and

far-infrared (FIR) spectrometry are quite different from those
discussed above for conventional, mid-IR spectrometry.

Near-infrared: 0.8 -2.5 m, 12500 - 4000 cm-1

Mid-infrared: 2.5 - 50 m, 4000 - 200 cm-1
Far-infrared: 50 - 1000 m, 200 - 10 cm-1

Divisions arise because of different optical materials and instrumentation.

IR alone cannot determine a structure.
Some signals may be ambiguous.
The functional group is usually indicated.
The absence of a signal is definite proof that the
functional group is absent.
Correspondence with a known sample’s IR
spectrum confirms the identity of the compound.

55
• Applications of IR :
• Applications of IR Fourier Transformed Infrared (IR) is important tool for

1. the solid state characterization of pharmaceutical solids and for the

identification of their chemical structures. Generally this method is applied
in combination with other methods for solid state characterization of
pharmaceutical solids (e.g. X- ray powder diffraction, DSC, TG).

2. characterization of polymorphism and detection in drug product. this

method is also suitable for the identification/ detection of the polymorphic
form in the tablet.

56
Advantages of FTIR :
1. Very high resolution required- Gaseous Mixtures

2. Study of samples having very high absorption

3. Study of samples with weak absorption bands

4. Used in protein structure determination

5. Very small sample size: Obtaining Reflection spectra IR

emission study.

Applications of ATR-FTIR :
• Surface analysis of biological structures
• Monitoring reaction processes
• Evaluation of fermented foods.

57
Pharmaceutical applications of Mid-IR
Mid-IR and Raman spectroscopy are versatile tools in pharmaceutics

and biopharmaceutics, with a wide field of applications ranging from

characterization of drug formulations to elucidation of kinetic

processes in drug delivery.

New developments in applications of these methods for studying drug

delivery systems.

FTIR-ATR is a well-established standard method used to study drug

release in semisolid formulations, drug penetration, and influence of

penetration modifiers; it is also capable of in vivo studies.

58
 Alzheimer’s Disease (AD)
• NIR FT-Raman spectra of control
and tumour brain tissue (white
and grey matter of normal brain
tissue identifiable).

• Raman spectra of AD brain tissue

show distinct differences from
normal tissue spectra

• Mizuno A, Kitajima H, Kawauchi K, Muraishi S and Ozaki Y

(1994) Near infrared Fourier transform Raman spectroscopic
study of human brain tissues and tumours J. Raman Spectrosc.
25 265–9

59
Pathology:
Breast implant Breast tumor

60
References:
1. Spectrometric Identification of Organic Compounds by Silverstein, Bassler
and Morrill;
2. Applications of absorption spectroscopy of organic compounds by Dyer JR.
3. Organic Chemistry by McMurry; 5thed; Thomason learning CA, USA 2000.
4. J. Workman, A.W. Springsteen, “Applied Spectroscopy”, Academic Press,
1998.
5. J.M. Hollas, “Modern Spectroscopy”, John Wiley&Sons, 1996.
6. B. Stuart, W.O. George, D.J. Ando, “Modern Infrared Spectroscopy”, John
Wiley&Sons, 1997.
7. N.N. Colthup, L.H. Daly, S.E. Wiberly, S.E. Wiberly, “Introduction to Infrared
and Raman Spectroscopy”, Academic Press, 1997.
8. B. Schrader, D. Bougeard, “Infrared and Raman Spectroscopy: Methods and
Applications”, John Wiley&Sons, 1995.