ANALYSIS BASED ON CHEMICAL CHARACTERIZATION METHOD (FTIR)

1. ANALYTICAL CHARACTERIZATION OF
BONE SCAFFOLD USING FTIR
• SUSHMITA DHIR

2. OBJECTIVES
Preparation of composite bone scaffold
FTIR spectra is performed to analyze the change in
chemical structure and characteristic wave number of
scaffold

3. LITERATURE SURVEY
SR
NO.
JOURNAL ( Author
and year of
publication)
TITLE INFERENCE
1. Srour, B., Bruechert,
S., Andrade, S. L. A.,
& Hellwig, P.
(2017).. Membrane
Protein Structure and
Function
Characterization,
195–203.
Secondary Structure
Determination by
Means of ATR-FTIR
Spectroscopy
Specialized infrared spectroscopic
techniques have been developed that
allow studying the secondary structure
of membrane proteins and the
influence of crucial parameters .
Hence infrared spectroscopy can be
used to identify the adapted sample
conditions.
2. Mudunkotuwa, I. A.,
Minshid, A. A., &
Grassian, V. H.
(2014). The Analyst,
139(5), 870–881.
ATR-FTIR spectroscopy
as a tool to probe
surface adsorption on
nanoparticles at the
liquid-solid interface in
environmentally and
biologically relevant
media
(i) The adsorption of different
environmentally and biologically
relevant coordinating ligands
(ii) Competitive ligand adsorption
(iii) The determination of kinetic and
thermodynamic parameters

4. 3. P, M., Rajakannu,
subashini, S, T.,
S, P., S, P. K., P,
B., &
Ponnusamy, S. K.
(2018). IET
Nanobiotechnolo
gy.
In vitro evaluation
of biodegradable
nHAPChitosan-
Gelatin-based
scaffold for tissue
engineering
application
FTIR was performed to identify important interactions
between natural polymer and bioactive component . the
functional groups PO43− and OH confirm the presence of
HAP nanoparticles, the functional groups Amide I, Amide II,
and P = O confirms the presence of chitosan nanoparticles
and the functional group CH confirms the presence of gelatin
in the scaffold. FTIR results clearly indicate the strong
bonding among chitosan, gelatin, and HAP nanoparticles
achieved by electrospun fabrication process.
4. Querido, W.,
Falcon, J. M.,
Kandel, S., &
Pleshko, N.
(2017). The
Analyst, 142(21),
4005–4017.
Vibrational
Spectroscopy and
Imaging:
Applications for
Tissue
Engineering
FTIR analysis can provide a rich and detailed range of
information using a nondestructive, label-free approach.
Spectroscopic analysis modalities are a compelling
alternative to standard evaluations due to their inherent
contrast-free nature, the ability to
investigate extracellular matrix and scaffold remodeling, and
the potential for non-destructive
evaluations in some applications.
5. Shai, Y. (2013).
Biochimica et
Biophysica Acta
(BBA) -
Biomembranes,
1828(10), 2306–
2313.
ATR_FTIR studies
in pore forming
and membrane
induced fusion
peptides
(i) Best choice to study the structure and organization of
membrane proteins and membrane-bound peptides in
biologically relevant membranes
(ii) Able to analyze material under a wide range od
conditions including solids, liquids, and gases.
(iii) Elucidation of component secondary structure elements
(iv) Determination of peptide orientation in the membrane,
secondary structures of very small peptides

5. DEFINITION
• Fourier-transform infrared spectroscopy (FTIR) is a technique used to
obtain an infrared spectrum of absorption or emission of a solid, liquid or
gas.
• An FTIR spectrometer simultaneously collects high-spectral-resolution
data over a wide spectral range.
Image source : Theophanides, T. (1984). Fourier transform infrared spectroscopy. Dordrecht: D. Reidel.
Fig 01 : Diagrammatic representation of the spectrometer

6. INTRODUCTION
• Infrared spectroscopy is the study of interactions between matter and
electromagnetic fields in the IR region.
• In this spectral region, the EM waves mainly couple with the molecular
vibrations.
• An infrared spectrum represents a fingerprint of a sample with absorption
peaks which correspond to the frequencies of vibrations between the bonds
of the atoms making up the material.
• In general, a frequency will be strongly absorbed if its photon energy
coincides with the vibrational energy levels of the molecule.
• IR spectroscopy is therefore a very powerful technique which provides
fingerprint information on the chemical composition of the sample.
• It can identify unknown materials.
• It can determine the quality or consistency of a sample.
• It can determine the amount of components in a mixture.

7. PRINCIPLE
• In FTIR analyses, Infrared light from the light source passes through a
Michelson interferometer along the optical path.
• As the moving mirror makes reciprocating movements, the optical path
difference to the fixed mirror changes, such that the phase difference
changes with time. The light beams are recombined in the Michelson
interferometer to produce interference light.
• The intensity of the interference light is recorded in an interferogram, with
the optical path difference recorded along the horizontal axis.
• Interferogram signal is converted to a frequency spectrum by a
mathematical technique called Fourier Transformation.

8. THE MICHELSON INTERFEROMETER
Image source : Delprat, P., & Gardette, J. (1993). Analysis of photooxidation of polymer materials by photoacoustic Fourier transform
infra-red spectroscopy. Polymer, 34(5), 933-937. doi: 10.1016/0032-3861(93)90210-2
Fig 02: Schematic diagram of a Michelson Interferometer.
• A broad-band light source
• A beam splitter
• Two front surface coated
mirrors – one moving and one
fixed
• A detector.

9. INSTRUMENTAL SETUP
1. The Source: Infrared energy is emitted from a glowing black-body source.
This beam passes through an aperture which controls the amount of energy
presented to the sample (and, ultimately, to the detector).
2. The Interferometer: The beam enters the interferometer where the “spectral
encoding” takes place. The resulting interferogram signal then exits the
interferometer. The interferometer uses a reference laser for precise
wavelength calibration, mirror position control and data acquisition timing.
3. The Sample: The beam enters the sample compartment where it is
transmitted through or reflected off of the surface of the sample, depending on
the type of analysis being accomplished. This is where specific frequencies of
energy, which are uniquely characteristic of the sample, are absorbed.

10. 4. The Detector: The beam finally passes to the detector for final measurement.
The detectors used are specially designed to measure the special interferogram
signal.
5. The Computer: The measured signal is digitized and sent to the computer
where the Fourier transformation takes place. The final infrared spectrum is
then presented to the user for interpretation and any further manipulation
Fig 03: Instrumental setup of FTIR
Image source : Introduction to Fourier Transform Infrared Spectroscopy by Thermo Fischer scientific corporation

11. SAMPLE PREPARATION
• In this work the sample is made in powder form. The powder
is compressed in pellet form.
• For mid‐IR frequency range, KBr, KCl or diamond dust can be
used. In far‐infrared testing the high‐density polyethene
(HDPE) or diamond dust is suitable. For near-infrared
analysis, CsI or KBr can be selected.
• The powder sample and KBr are grounded to reduce the
particle size less than 5 mm in diameter.
• A good KBr pellet is thin and transparent.

12. SPECTRUM ANALYSIS OF SAMPLE
Fig 1. FTIR Spectra of sample with guar gum
Fig 2FTIR spectra of sample with xanthan gum

13. Table 01 : Characteristic group and IR wave
number of guar gum and xanthan gum.

14. REFERENCES
1. Querido, W., Falcon, J. M., Kandel, S., & Pleshko, N. (2017). Vibrational
spectroscopy and imaging: applications for tissue engineering. The Analyst, 142(21),
4005–4017. doi:10.1039/c7an01055a
2. P, M., Rajakannu, subashini, S, T., S, P., S, P. K., P, B., & Ponnusamy, S. K. (2018).
In vitro evaluation of biodegradable nHAP-Chitosan-Gelatin based scaffold for
tissue engineering application. IET Nanobiotechnology. doi:10.1049/iet-
nbt.2018.5204
3. Mudunkotuwa, I. A., Minshid, A. A., & Grassian, V. H. (2014). ATR-FTIR
spectroscopy as a tool to probe surface adsorption on nanoparticles at the liquid–
solid interface in environmentally and biologically relevant media. The Analyst,
139(5), 870–881. doi:10.1039/c3an01684f
4. Srour, B., Bruechert, S., Andrade, S. L. A., & Hellwig, P. (2017). Secondary Structure
Determination by Means of ATR-FTIR Spectroscopy. Membrane Protein Structure
and Function Characterization, 195–203. doi:10.1007/978-1-4939-7151-0_10
5. Shai, Y. (2013). ATR-FTIR studies in pore forming and membrane induced fusion
peptides. Biochimica et Biophysica Acta (BBA) - Biomembranes, 1828(10), 2306–
2313. doi:10.1016/j.bbamem.2012.11.027
6. : Delprat, P., & Gardette, J. (1993). Analysis of photooxidation of polymer materials
by photoacoustic Fourier transform infra-red spectroscopy. Polymer, 34(5), 933-937.
doi: 10.1016/0032-3861(93)90210-2