Chemistry for BME
Chapter 6: Spectroscopic Methods
Part 3: Fourier-Transform Infrared Spectroscopy
Instructor: Dr. Vu Thanh Binh
Email: vtbinh@[Link]
Chapter 6 – Part 3 Infrared Interactions
How electromagnetic radiation interacts with matter?
Radiation Type Main Interaction
X-ray Ionization
UV-Vis Electronic excitation
Infrared Molecular vibration
Microwave Molecular rotation
Nuclear spin
Radiofrequency
transitions
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Chapter 6 – Part 3 Infrared Interactions
>14,000 cm-1 14,000 to 4000 cm-1 4,000 to 400 cm-1
Visible & UV Near IR Mid-Infrared
Electronic Transitions Molecular Vibrations Molecular Vibrations
400 to 4 cm-1 <4 cm-1
Far Infrared Microwaves
Molecular Molecular
Vibrations Rotations
Higher Wavenumber Lower Wavenumber
Higher Frequency Lower Frequency
Higher Energy Lower Energy
Shorter Wavelength Longer Wavelength
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Chapter 6 – Part 3 Infrared Interactions
• The quantum energy of infrared photons is in the range 0.001 to 1.7 eV which is in the
range of energies separating the quantum states of molecular vibrations.
• Infrared is absorbed more strongly than microwaves, but less strongly than visible light.
• The result of infrared absorption is heating of the tissue since it increases molecular
vibrational activity. Infrared radiation does penetrate the skin further than visible.
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Chapter 6 – Part 3 Infrared Interactions
• A molecular vibration occurs when atoms in a molecule are in periodic motion while the
molecule as a whole has constant translational and rotational motion.
• The frequency of the periodic motion is known as a vibration frequency, and the typical
frequencies of molecular vibrations range from less than 1013 – 1014 Hz approximately,
corresponding to wavenumbers of approximately 300 to 3000 cm-1.
Vibrational modes leading to IR absorptions:
Stretching Vibrations
A stretching vibration changes the bond length. There are two types:
Asymmetric Stretching
Symmetric Stretching Where some bonds are getting
Where two or more bonds shorter at the same time as
vibrate in and out together. others are getting longer.
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Chapter 6 – Part 3 Infrared Interactions
Bending Vibrations
(Deformation Vibrations, changing
the angle between two bonds)
In-plane:
Rocking is like the motion of
Scissoring is the movement a pendulum on a clock, but an
of two atoms toward and atom is the pendulum and
away from each other. there are two instead of one.
Out-of-plane:
Wagging is like the motion in which
Twisting is a motion as if you make a “V” sign with your
the atoms were walking on fingers and bend them back and
a treadmill. forth from your wrist. 6
Chapter 6 – Part 3 Sample Handling
v Solid Sampling Technique
Preparing solid samples:
a. Direct sampling: the solid sample is directly placed in a sample holder.
b. Pelletization technique: the solid sample is mixed with KBr and then passed under very high
pressure in a press to form small 1-2 mm thick pellets (1 cm diameter). These pellets are
transparent to IR radiation. For IR spectroscopy only KBr pellets than KBr+ drug pellets.
c. Mulling techniques: the solid sample is mixed with heavy mineral oil (Nujol) to form a paste,
which is placed between two salt plates. Nujol is transparent in most of the IR region. Other
mulling agents are CFC, hexachlorodiene, etc.
d. As solid film: the sample solution is placed on a KBr or NaCl surface and the solvent is
evaporated. On the surface of the cell, the solid sample consequently leaves behind a thin layer.
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Chapter 6 – Part 3 Sample Handling
v Liquid sample
A liquid sample is squeezed or sandwiched between two sodium chloride plates and
forms a thin film (0.10.3 mm). Sodium chloride is transparent to IR. Calcium fluoride
plates are used for water-containing samples.
v Gas sample
A gaseous sample is placed into a gas cell (10 cm) having sodium chloride walls (NaCl
allows IR).
v Solution sampling technique
The sample is dissolved with various solvents and then analyzed in the form of a
solution. (E.g. Chloroform, Carbon Tetrachloride, Carbon Disulfide)
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Chapter 6 – Part 3 Fourier Transform Spectrometers
How to make a KBr pellet?
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Chapter 6 – Part 3 Instrumentation
It is used to measure the intensity of the sample in IR spectroscopy.
The main components of an IR spectrometer are:
• Source Radiation
• Monochromators
(Wavelength selector)
• Sample cells
• Detectors
• Recording system
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Chapter 6 – Part 3 IR Spectroscopy
Infrared spectrum represents a fingerprint of a sample with absorption peaks corresponding to the
frequencies of vibrations between the bonds of the atoms making up the material.
• Infrared spectroscopy can provide identification (qualitative analysis) of every different kind
of material.
• The size of the peaks in the spectrum is a direct indication of the amount of material present,
providing quantitative analysis.
Excitation of carbonyl bond in 2-hexanone 11
Chapter 6 – Part 3 IR Spectroscopy
• Absorption occurs when IR frequency
matches the natural vibrational
frequency of a bond.
• Molecules exhibit different vibrations:
bond stretching, bond bending.
• Different functional groups absorb at
characteristic IR frequencies: C=O
(carbonyl), C–H, C–C, O–H, N–H, etc.
• The resulting IR spectrum acts as a
IR spectrum of 2-hexanone
molecular fingerprint.
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Chapter 6 – Part 3 IR Spectroscopy
Fingerprint region
• The blue-coloured line - stretching vibrations
• The green-coloured line - bending vibrations
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Chapter 6 – Part 3 IR Spectroscopy
Example:
Identify the various vibrations in the infrared spectrum of methanal (formaldehyde), H2C=O, given the
following data:
• 1750 cm-1
• 2785 cm-1
• 2850 cm-1
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Chapter 6 – Part 3 IR Spectroscopy
Example:
Identify the various vibrations in the infrared spectrum of methanal (formaldehyde), H2C=O, given the
following data: The fingerprint region from 0 to 1500 cm-1 is not
Solution: typically analyzed.
• 1750 cm-1
• 2785 cm-1
• 2850 cm-1
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Chapter 6 – Part 3 Dispersive Infrared Instruments
Diagram of a multichannel spectrometer based on a grating spectrograph with a photodiode array detector.
In ultraviolet/visible instruments, cells are always located between the monochromator and the
detector in order to avoid photochemical decomposition. In contrast, Infrared radiation is not
sufficiently energetic to bring about photodecomposition; thus, the cell compartment can be
located between the source and the monochromator.
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Chapter 6 – Part 3 FTIR Spectrometer
Illustration of FTIR spectrometer
• Fourier Transform Infrared (FTIR) spectrometry is designed to overcome the
limitations of dispersive equipment.
• It is preferable to measure all IR frequencies at the same time
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Chapter 6 – Part 3 Fourier Transform Spectrometers
Fourier transform infrared (FTIR) spectrometers offer the advantages of high sensitivity,
resolution, and speed of data acquisition. It utilizes an ingenious device called a Michelson
interferometer.
• IR radiation is divided by a beam
splitter and reflected by both a fixed
mirror and a movable mirror.
• When both mirrors are at equal
distances, the optical path difference Fixed mirror = Movable mirror?
is zero.
• The reflected beams recombine and
pass through the sample to the
Instrumentation of FTIR 18
detector.
Chapter 6 – Part 3 Fourier Transform Spectrometers
• As the movable mirror changes position, the path lengths become different, creating
an optical path difference.
• This produces constructive or destructive interference in the interferogram.
• The movable mirror position is precisely monitored using a helium–neon laser.
(Optical Path Difference) 19
Chapter 6 – Part 3 Fourier Transform Spectrometers
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Chapter 6 – Part 3 Fourier Transform Spectrometers
• Constructive Interference: Waves with matching crests and troughs combine to produce
a larger amplitude.
• Destructive Interference: Waves with opposite phases (crest and trough) cancel each
other, resulting in reduced or nearly zero amplitude.
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Chapter 6 – Part 3 Fourier Transform Spectrometers
The beam from the moving mirror travels at a different distance than the beam from
the fixed mirror. Interference pattern is created when the beams are combined.
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Chapter 6 – Part 3 Fourier Transform Spectrometers
Detector
• A device that detects the absorbance of the beam through the sample is called detector.
• The detectors used in FTIR spectrophotometer should be such that it is speed and
accuracy matches with the speed and sensitivity of the instrument.
• These detectors are classified into two categories:
Fourier transformation
Ø Thermal detector
Translated
Ø Quantum mechanical detector
by FT
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Chapter 6 – Part 3 Fourier Transform Spectrometers
Detector
Comparison
Thermo Detector Quantum Mechanical Detector
• A thermo detector uses the infrared beam as • A quantum mechanical detector uses the IR
heat. beam as light and provide for more sensitive
• Operates by detecting changes in temperature detector.
of an absorbing material.
Example Example
Pyroelectric detector Photon-sensitive semiconducting detector
Deuterated Tryglicine Sulphate (DTGS) Mercury Cadmium Telluride (MCT)
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Chapter 6 – Part 3 Fourier Transform Spectrometers
• In FTIR the change in signal is measured with time ® time domain spectra.
• Cannot deduce frequency-domain spectra from the corresponding time-domain
waveform ® need to perform Fourier transform.
• Signal obtained from the many wavelengths in the (polychromatic) source is a
direct function of the optical path difference or retardation (δ):
Where:
Ø 𝐼(𝛿) - intensity at the detector
Ø B(𝜈) - beam spectrum, or the intensity at wavenumber (𝜈)
Ø δ - optical path difference or retardation
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Chapter 6 – Part 3 Fourier Transform Spectrometers
Interferogram Spectrum
a simple sinusoidal interferogram a single sharp peak
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Chapter 6 – Part 3 Attenuated Total Reflection FTIR (ATR-FTIR)
FTIR Sample Techniques
• In the early days of infrared spectroscopy, the only available method of analysis was
transmission.
• There are four major sampling techniques in FTIR:
Ø Transmission - FTIR
Ø Attenuated Total Reflection (ATR) – FTIR
Ø External Reflection - FTIR
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Chapter 6 – Part 3 Fourier Transform Spectrometers
Most traditional FTIR Useful for analyzing coatings
Widely used because it requires Suitable for solids, liquids, and
minimal sample preparation biological samples
Three typical modes of FTIR: (a) transmission, (b) external reflection and (c) ATR modes.
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Chapter 6 – Part 3 Fourier Transform Spectrometers
What is Attenuated Total Reflectance (ATR)?
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Chapter 6 – Part 3 Fourier Transform Spectrometers
How to perform FTIR instrument?
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Chapter 6 – Part 3 Attenuated Total Reflection FTIR (ATR-FTIR)
ADVANTAGES DISADVANTAGES
Fast and easy sample analysis quantitation It lacks sensitivity when compared with other techniques
Analysis of different types of samples The ATR accessory is costly
Non-destructive technique Crystal care should be taken in case of crystals like ZnSe
Straight forward Limited measuring wave number range
Crystal material Refractive index Wave number range(cm-1) Color pH range
KRS-5 2.37 20,000–250 Red 5–8
ZnSe 2.42 15,000–600 Yellow 5–9
Si 3.42 8900–660 Grey 1–12
Ge 4.0 5500–600 Grey
Diamond 2.42 30,000–2200, 2000–400 Clear 1–14
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Chapter 6 – Part 3 Quantitative Infrared Spectrometry
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Chapter 6 – Part 3 FTIR Spectral Interpretation
Classification of IR Bands
IR bands can be classified as strong, medium or weak, depending on their relative intensities
in the infrared spectrum.
Ø A strong band covers most of the y-axis.
Ø A medium band falls to about half of the
y-axis.
Ø A weak band falls to about one third or
less of the y-axis.
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Chapter 6 – Part 3 FTIR Spectral Interpretation
• Not all covalent bonds display bands in the IR spectrum. Only polar bonds do so. These are
referred to as IR active. There will be a change of dipole moment if the atoms at each end of its
chemical bond are not the same element. So H-Cl is IR active; but Cl-Cl and H-H are silent in the
infrared region of the electromagnetic spectrum.
• The intensity of the bands depends on the magnitude of the dipole moment associated with the
bond in question:
Ø Strongly polar bonds such as carbonyl groups (C=O) produce strong bands.
Ø Medium polarity bonds and asymmetric bonds produce medium bands.
Ø Weakly polar bond and symmetric bonds produce weak or non observable bands.
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Chapter 6 – Part 3 FTIR Spectral Interpretation
Infrared Band Shapes
IR band shapes come in various forms. Two of the most common are narrow and broad.
• Narrow bands are thin and pointed, like a dagger.
• Broad bands are wide and smoother.
A typical example of a broad band is that displayed by O-H bonds, such as those found in
alcohols and carboxylic acids, as shown below.
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Chapter 6 – Part 3 FTIR Spectral Interpretation
• The typical IR absorption range for covalent bonds is 600 – 4000 cm-1.
• The graph shows the regions of the spectrum where the following types of bonds
normally absorb.
• For example, a sharp band around 2200 – 2400 cm-1 would indicate the possible
presence of a or a .
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Chapter 6 – Part 3 FTIR Spectral Interpretation
Difference between IR and FTIR?
• IR stands for Infrared spectroscopy and FTIR stands for Fourier Transform Infrared
spectroscopy. IR spectroscopy and FTIR spectroscopy are related to the infrared region of
the electromagnetic spectrum, which is light with a long wavelength and low frequency
compared to visible light.
• It covers many techniques, which is based mostly on absorption spectroscopy. Like all
spectroscopic techniques, it can be used to identify and study of analytes or components.
• IR spectroscopy is a significant technique in organic chemistry. This is a simple technique to
identify the presence of functional groups in a molecule.
• In addition, to verify the identity of a pure compound or specific impurities, the absorption
can utilize the unique collection of the band.
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Chapter 6 – Part 3 FTIR Spectral Interpretation
Difference between IR and FTIR? (continue)
Ø IR spectroscopy gets absorption of a monochromatic IR light at a time and draws the spectrum,
whereas, in FTIR, Multi-chromatic takes a summarized absorption of light and distributes it to create a
spectrum using FTIR. This is faster and more effective technique.
Ø Here are some points of difference between IR and FTIR.
• IR takes a single spectrum • FTIR employs an interferometer and takes a numbers of scans
• IR used monochromatic light whereas • FTIR scans up to 50 times in a minutes and giving better
FTIR used polychromatic light. resolution.
• Time-consuming • All analytes can be identified with a single measurement and
the interferences are resolved.
• FTIR is a fast technique than IR
• No time-consuming to prepare the sample
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Chapter 6 – Part 3 FTIR Analysis
Visualization of a chemical reaction
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Chapter 6 – Part 3 FTIR Analysis
Determination of a chemical
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Chapter 6 – Part 3 Applications of FTIR
Applications of FTIR
• Atmospheric & Environmental Monitoring
Detection of gases such as CO₂, CH₄, and ozone
• Food & Industrial Analysis
Rapid compositional analysis of oils, fats, and food products
• Materials Science & Polymers
Analysis of crystallinity, oxidation, degradation, and surface chemistry
• Biomedical & Pharmaceutical Applications
Protein structure analysis, biomaterials, and diagnostics
• Geochemical & Catalytic Studies
Investigation of mineral composition and adsorption processes
• Biophysical Studies
Detection of structural changes in biomolecules and photosynthetic systems
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