Introduction to FT-IR Spectroscopy

July 2, 2012 1
Contents

• Infrared Spectroscopy
• Molecular Vibration
• FT-IR Spectrometer
• Interferometer
• Fourier Transformation (FT)
• Measurement Techniques
ATR: Attenuated Total Reflection
• Evaluation of IR-Spectra

2
Infrared Spectroscopy

3
Electromagnetic radiation

4
What is IR-Spectroscopy

Interaction between light and matter

• Absorption of IR light by sample

• Excitation of vibrations
5
Molecular Vibration

6
 Interaction of radiation and matter

Vibration theory
IR spectroscopy is based on the absorption of infrared light by the substance to be measured. This absorption excites molecular
vibrations and rotations, which have frequencies that are the same as those within the infrared range of the electromagnetic
spectrum.

The following simple model of a harmonic oscillator used in classical physics describes IR absorption. If atoms are considered to
be particles with a given mass, then the vibrations in a diatomic molecule (e.g. HCl) can be described as follows:

Mechanical model of a vibrating diatomic molecule

7
 Vibration theory

An accurate model of a molecule is given by the

anharmonic oscillator. The potential energy is then
calculated by the Morse equation, and is asymmetric. The
energy levels are no longer equally spaced, and are given
by:

Ev=(v + ½) h   - (v + ½)2 xGl h  

where xGl is the anharmonicity constant.

The anharmonic oscillator model allows for two important

effects:
1) As two atoms approach each other, the repulsion
increases very rapidly.
2) If a sufficiently large vibrational energy is reached the
molecule will dissociate (break apart). This is called the
dissociation energy.

In the case of the anharmonic oscillator, the vibrational

transitions no longer only obey the selection rule v = 1.
This type of vibrational transition is called fundamental Potential energy curve for an anharmonic oscillator
vibration. Vibrational transitions with v = 1, 2, 3, ...
are also possible, and are termed overtones.

8
Which kind of vibrations?

For example: water

Stretching vibrations

Symmetric stretching vibration Anti-symmetric stretching vibration

9
Which kind of vibrations?

Deformation vibrations

10
IR-Spectrum of Water

100
95 85 90
Transmission [%]
75 80
70
65
60

3500 3000 2500 2000 1500

wavenumber cm-1
11
IR-Spectrum of Hexane

n-Hexane

More than 50 different vibrations

12
 IR-Spectrum of Hexane
100
80
Transmission [%]

C-H stretch C-H deformation

40 20 60

„Fingerprint region“

4000 3500 3000 2500 2000 1500 1000

wavenumber cm-1

13
 IR radiation is devided into
3 spectral ranges

NIR MIR FIR

15.000 cm-1 4.000 cm-1 400 cm-1 5 cm-1

14
 Spectral ranges:
Near-(NIR), Middle-(MIR), Far-(FIR)Infrared

Characterization of spectral ranges:

NIR: 4000 - 15.000 cm-1
Overtones and combination vibrations
low absorption coefficient <--> high sample concentrations
Advantage: Quartz is transparent --> fibre optics, in glass vials

MIR: 400 - 4000 cm-1

Fundamental molecular vibrations: stretch and deformation vibrations
high absorption coefficient <--> low sample concentrations

FIR: 5 - 400 cm-1

backbone vibrations of large molecules, molecules with heavy atoms
low absorption coefficient, strong water vapor absorption --> vacuum spectrometer

15
Differences between NIR, MIR, FIR

Optical components:

NIR: Source: tungsten lamp

Optical material: Quartz
Detector: InGaAs, InSb

MIR: Source: Globar

Optical material: KBr, ZnSe
Detector: DTGS, MCT

FIR: Source: Globar, Hg lamp

Optical material: PE, CsI
Detector: FIR-DTGS, Bolometer

16
FT-IR Spectrometer

17
Principle of Infrared spectroscopy

Excitation of molecular vibrations,

Spectral analysis

18
Principle layout of IR spectrometers

Dispersive IR spectrometer FT-IR spectrometer

19
Layout of an FT-IR spectrometer

Electronic system Source compartment

Detector
compartment

Aperture &
Sample compartment Filter wheel

Sample position

Interferometer compartment

20
Michelson Interferometer

Fixed
mirror

Moving
mirror
L
x Source

L + x

x=0 Beamsplitter

Detector

21
22
 Origin of the Interferogram

IR light source Resulting detector signal

Intensity

Intensity

Frequency

X, moving mirror

23
 Transmission Spectrum

Detector intensity
1.) In the empty sample compartment an
Interferogram is detected. The result
of the FOURIER transformation is R(ν).

X, moving mirror

Fourier-Transformation

Single channel intensity

0.10 0.20 0.30 0.40

4000 3500 3000 2500 2000 1500 1000 500

wavenumber cm-1
24
 Transmission Spectrum

Detector intensity
2.) A second interferogram is detected with
the sample placed in the sample
compartment. The result of the FOURIER
transformation is S(ν). S(ν) shows
similarities to the reference spectrum
R(ν), but has lower intensities at the
regions the sample absorbs radiation.
X, moving mirror
Fourier-Transformation

Single channel intensity

0.40
0.30
0.20
0.10

4000 3500 3000 2500 2000 1500 1000 500

wavenumber cm-1
25
 Transmission Spectrum

3.) The transmission spectrum T(ν) is

0.10 0.20 0.30 0.40

Single channel intensity
calculated as the ratio of the sample
and reference single channel spectra:

T(ν) = S(ν) / R(ν)

ratio

100
Transmission [%]
40 60 80
20

4000 3500 3000 2500 2000 1500 1000 500

wavenumber cm-1

26
Fourier Transformation (FT)

27
The Fourier Transform

Data acquisition results in a digitized interferogram, I(x),

which is converted into a spectrum by means of the
mathematical operation called a Fourier Transform (FT).

The general equation for the Fourier Transform is

applicable to a continuous signal. If the signal
(interferogram) is digitized, however, and consists of N
discrete, equidistant points, then the discrete version of
the FT (DFT) must be used:

S(k . Δ ~
) = Σ I(n  Δx)  exp (i2πk  n/N)

The continuous variables x and ~  have been replaced

with n  Δx and k  Δ~  , representing the n discrete
interferogram points and the k discrete spectrum points.
The fact that we now have a discrete, rather than
continuous, function, and that it is only calculated for a
limited range of n (i.e. the measured interferogram has a
finite length) leads to important effects known as the
picket-fence effect and leakage.

28
Spectral Resolution

d IR Spectrum

Two separated lines with the

distance d in the spectrum lead to a wavenumber cm-1

periodic signal in the interferogram

with a distance 1/d.

Interferogram
RAYLEIGH criterion:

x  1/d

29
 Spectral Resolution

High spectral resolution

6595.5 6595.0 6594.5 6594.0 6593.5 6593. 0 6592.5 6592.0
Wavenumber cm-1

fixed
mirror M1
movable
mirror M2
L
x source

6595.5 6595.0 6594.5 6594.0

Wavenumber cm-1
6593.5 6593. 0 6592.5 6592.0

L + x
C:\Programme\OPUS5-finalrelease\DATA\Crnin2ob.0 N2O LP-GAS-CELL 24/01/89

Beam splitter
x=0

detector

Low spectral resolution

30
 Absorbance <-> Transmission - Why?

1.0
100

Transmission Absorbance

0.8
80
Transmittance [%]

Absorbance Units
TR = I / Io Lambert-Beer‘s law:

0.6
60

AB = -log (I / Io)
AB =  • c • b

0.4
40

fine features can be better AB prop. c

0.2
20

observed (linear y-scale) Quantification

than in AB
0.0
0

6000 5000 4000 3000 2000 1000 6000 5000 4000 3000 2000 1000
Wavenumber cm-1 Wavenumber cm-1

31
Measurement Techniques

ATR
(Attentuated Total Reflection)

32
ATR Theory

sample

n1 > n 2 !

sample
IR beam ATR crystal
n2
n1
Penetration Depth: θi
ATR crystal
 l : wavelength
dp  n1 : refractive index of crystal
2 n1 sin 2  n21
2
 : angle of incidence
n21: ratio of refractive index sample/crystal
Harrick approximation
33
 Penetration Depth

Calculated penetration depth for typical ATR materials

The penetration depth Dp depends on the

following parameters: Refractive index
Dp * Dp *
Material
1.) angle of incidence @ 1.000 cm-1 [µm] @ 45° [µm] @ 60°

2.) refractive indices of crystal and sample

Diamant 2.4 1.66 1.04

3.) wavelength of light

Ge 4 0.65 0.5
4.) contact between crystal and sample
Si 3.4 0.81 0.61
5.) surface of sample (rough/smooth)

6.) number of reflections in crystal ZnSe 2.4 1.66 1.04

7. (polarization of light)
AMTIR** 2.5 1.46 0.96

* : calculated for a sample with n = 1.4 @ 1000cm-1

** : AMTIR: Ge33As12Se55 - Glass

34
Penetration Depth

wavelength

2 µm 25 µm

dp
dp
sample

0.0 0.2 0.4 0.6 0.8 1.0

dP prop  Transmission
ATR
Absorbance

3500 3000 2500 2000 1500 1000

wavenumber cm-1

35
Evaluation of IR-Spectra

36
 Evaluation of Spectra

Infrared spectroscopy is an extremely efficient analytical

method due to modest operating expenditure. The
analytical results are provided within a short period of time
without the need of extensive sample preparation. In
particular, infrared spectroscopy provides data which can
be evaluated by quantity as well as by quality. The
following will describe the qualitative and quantitative
evaluation of acquired spectra.

• Qualitative evaluation of spectra

1. Identify an unknown substance

2. Check the identification of a known substance

• Quantitative evaluation of spectra

37
 Qualitative Evaluation:
Identification of Unknown Substances

a) Structural determination by interpreting spectra

A functional group within a molecule is considered as a harmonic oscillator (see vibration theory) which in a first approximation
vibrates without being affected by the rest of the molecule. This results in the fact that a particular functional group shows IR
absorption bands within characteristic spectral ranges: this is called group vibrations. This fact serves as the basis for spectral
interpretation, whereby the position, (relative) intensity and half-width of a band decide whether a band can be assigned to a
specific structural group.

Many functional groups of organic molecules show characteristic vibrations corresponding to absorption bands within defined ranges
of the IR spectrum. These molecular vibrations are mainly restricted to the functional group and do not affect the remaining
molecule, i.e. such functional groups can be identified by their absorption band. This circumstance, apart from a straightforward
acquisition technique, makes IR spectroscopy to be one of the simplest, fastest and most reliable methods when assigning a
substance to its specific class of compounds. The position and intensity of the absorption bands are extremely specific in the case of
a pure substance. This enables the IR spectrum, similar to the human fingerprint, to be used as a highly characteristic feature for
identification.
100 80
Transmission [%]
40 60
20

4,000 3,500 3,000 2,500 2,000 1,500 1,000 500

Wavenumber / cm-1

38
IR absorption of different
organic molecular classes

39
Qualitative Evaluation:
Identification of unknown Substances

b.) Comparing with spectral libraries

Besides basic spectral interpretation, various
comprehensive digital spectral libraries have been compiled
according to different chemical classes and groups of
substance. These are provided, for example, by companies
like Bruker and Sadtler. Apart from working with existing
spectral libraries, it is possible to create your own libraries
using modern spectroscopic software,see OPUS/SEARCH.
Different spectra regarding the number of bands and half-
width, may require different search algorithms. Therefore,
OPUS/SEARCH has the flexibility in providing various
search options.

40
 Qualitative Evaluation:
Check the Identity of known Substances

Infrared spectroscopy is a perfect analytical tool for quality

control. It gives the answer to the following question: “Does
the quality of the raw material delivered to the receiving
department comply with the specifications?” The underlying
concept is very easy:
identical material = identical IR spectrum
The identification is done by comparing measured spectra
with reference spectra already saved. The method is based
upon the following considerations:
• chemically different materials result in different
spectra
• real spectral differences exceed the reproducibility
of repeated measurements
• reference samples represent the expected sample
variations caused by supplier, batch, season,
purity, grain size etc.
It is important to note that the reference samples can vary
to a certain degree, a circumstance that is experienced
within quality control every day. The spectrum of the
material to be identified is compared with the reference
sample by means of a valid tolerance previously defined.
How to create a reference library and to compare spectra
will be described in the following.

41
Qualitative Evaluation:
Check the Identity of known Substances

Compare spectra

42
 Reference library structure

1.) Measure reference 2.) Calculate average spectrum 3.) Library structure
samples & threshold values & validation
Absorbance


  
Absorbance

   
  
  
   
 
 
  


Wavenumber / cm-1
Wavenumber / cm-1

43
Identifying new samples

1.) Measure new samples 2.) Compare with library 3.) Identify material

Identified
sample:
material X

44
 Quantitative Evaluation of Spectra

The basic principle for quantitative evaluation in optical

spectroscopy as well as in IR spectroscopy is the Bouguer-
Lambert-Beer Law which had already been defined in 1852.
Quantitative determinations by means of IR spectroscopy are
preferably performed in solution. Transmission T of a sample is
defined as:

T = I / I0

Io is the intensity of the incident light beam, I is the intensity of

the light beam leaving the sample. The percentage transmission
(%T) is 100 • T. When traversing the measurement cell, the light
intensity decreases exponentially:

I = I0 • exp(-2.303 • c • b)

Where  is the molar absorption coefficient (in L mol-1 cm-1), c

is the sample concentration (in mol L-1) and b the thickness of
the measurement cell (in cm). The absorption coefficient  is a
value which depends on either the wavelength or the
wavenumber, which is typical for the compound analyzed. From
the equation above, it follows that:

log (I / I0) = - • c • b, or:

A = log (I0 / I) =  • c • b

where A is the absorbance. Because of the Bouguer-Lambert-

Beer Law, the relationship between absorbance and
concentration of the absorbing substance is a linear function.

45
Quantitative Evaluation of Spectra

In practice the relationship between concentration and Calibration

absorbance is empirically determined by calibration.
Calibrating means finding the mathematical connection 4
between concentration and the measurement values.

Absorbance
3
In the first step, spectra of substances with known
composition are recorded. Then, these acquired spectra X
and the data available from a reference analysis
(concentration or substance properties) are used to 2
determine a calibration function. The software package
OPUS/QUANT provides several algorithms to do this.
1
In the second step, spectra of substances with an unknown
composition are measured and then used to determine the
properties of interest by means of the calibration function. Wavelength
There are two different forms of calibration:
Analysis
Univariate calibration (OPUS) 4
Correlates just one piece of spectral information (e.g. peak

Absorbance
height or peak area) with the reference values of the
3
calibration set.
X
Multivariate calibration (OPUS/QUANT)
2
Correlates considerably more spectral information using
larger spectral ranges with the reference values of the
calibration set. This leads to a higher degree of precision 1
and reduced chance of error. Partial Least Squares ((PLS)
is an example of this method and is implemented in
OPUS/QUANT. Concentration

46
 If you have any
further questions
about IR spectroscopy,
please contact Bruker
Optics’ application
team:

www.brukeroptics.com

North America: Europe: Asia:

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Phone: +1 978 439 9899 Phone: +49 7243 504 600 No. 9, Sheung Yuet Road, Hong Kong
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info@brukeroptics.com info@brukeroptics.de Fax: +852 4927966109
asiapacific@brukeroptics.com.hk

47
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