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The Hong Kong Polytechnic University

Department of Applied Biology and Chemical Technology

ABCT458
Advanced Analytical Techniques Lab
Report

Experiment: FTIR
Reflectance Measurements
Using A FTIR Spectrophotometer

Mar 25, 2008

Group 1B
Name (Student Number) Duty
NI Qing (05995048d) Group Leader
MAN Lai Fan (07515458d) Introduction
YEUNG Shing Hin (07533415d) Theoretical Section
LI Lai Ping (07546484d) Experimental Section
FUNG Yat Sing (07524431d) Results & Discussion

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Table of Contents
1. Introduction........................................................................................................................................................1

2. Theory................................................................................................................................................................2

A. Attenuated Total Reflection (ATR)...................................................................................................................2

B. Diffuse Reflectance Infrared Fourier Transform Spectroscopy (DRIFT).........................................................2

3. Procedure............................................................................................................................................................2

Part 1.................................................................................................................................................................2

Part 2.................................................................................................................................................................3

4. Results and Discussion.......................................................................................................................................3

(1) Comparison of the Milk Contents (ATR).......................................................................................................3

(2) Identification of Unknown Polymers with DRIFT spectroscopy....................................................................4

5. Conclusion..........................................................................................................................................................7

6. References..........................................................................................................................................................7

7. Appendices.........................................................................................................................................................8

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1. Introduction
Fourier transform infrared spectroscopy (FTIR) is a measuring technique of
getting the infrared spectra by measuring the interferogram (time-domain signal) of the
samples with the use of interferometer and then the interferogram is Fourier transformed
to give the spectrum (frequency-domain functions). Fourier transform is a data processing
tool and infrared spectroscopy is the study of interaction of infrared radiation with matter
with the resultant spectrum showing the absorption due to various distinctive functional
groups, giving a molecular fingerprint of the studied compound. FTIR can be applied for
the identification of the unknown, determination of the quality of the sample and the
components of a mixture.
There are mainly two common types of infrared sampling techniques-they are the
transmission techniques and the reflectance techniques. For the transmission mode, the
infrared radiation beam passes through the sample directly. And it is the most popular
way of obtaining the infrared spectra because of its various advantages such as it is
relatively inexpensive, having high signal to noise ratio and universal –can work on
various kinds of samples such as polymers, gases, liquids and solids that can be ground
into a powder with alkaline metal halide such as KBr pellet or suspended in an oil. In the
reflectance technique, the infrared light beam is bounced off the sample rather than
passing through the sample. The reflectance technique can solve the thickness problem
that the transmission mode may have. Hence, the method is less time consuming as there
is no need for the sample to be in the proper concentration, thickness or the proper
amount of light passing through and hence no dilution or grinding of the sample is
required. And the method is non-destructive. Hence, the technique can be applied to case
where it is difficult to study with the transmission technique. The attenuated total
reflection (ATR) and the Diffuse reflectance infrared Fourier transform spectroscopy
(DRIFTS) are studied in this experiment. DRIFTS can be used to examine powder and
solid, even very large and intractable samples such as pieces of plastic and furniture.
ATR can be applied to examine polymer films and semisolid samples such as shampoo,
toothpaste and tomato sauce etc.

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In this experiment, various milk and polymer samples are studied with the reflectance
measurement techniques using the FTIR spectrophotometer.[1-3]

2. Theory

A. Attenuated Total Reflection (ATR)


There is a phenomenon called total reflection that a light beam will be totally
reflected when it travels from a more refractive medium to less refractive one with
incident angel greater than critical angel.

Critical angel, θ c = sin − 1 η 2 where is η1 the refractive index of more refractive


η1
medium and η2 is the refractive index of less refractive medium.

In ATR, a highly refracting prism is used. Although the IR light beam is expected
totally refracted, evanescent wave will get through to the shallow surface of the less
refractive medium and absorbed by the sample that contact to the prism.

B. Diffuse Reflectance Infrared Fourier Transform Spectroscopy (DRIFT)


Diffuse reflection and specular reflection are the components observed of
reflection from solid surface. In DRIFT, light shines onto the powdered sample, specular
component, where the light is reflected directly off, was blocked and diffuse reflectance,
where the light penetrated, transmitted and reflected within the sample, was collected and
analyzed.

Both of the methods can use in sample that is not pulverized or palletized easily
which cause difficulty in analysis by typical methods.[1-3]

3. Procedure

Part 1
(The power of the PC had turned on and the spectrometer operating software had loaded)

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The parameters such as resolution were set on the spectrometer operating software. We
had made sure that the surface of the ATR prism was cleaned. The background was then
collected by operating the spectrometer. After that, a few drops of water were added on
the surface of the prism by dropper. The spectrum of water was collected. The water was
removed from the prism surface by absorbing it away with soft tissue paper. The full-
cream milk samples were then added to the surface of prism by the same steps of adding
water. And the spectrum of full-cream milk was collected by operating the spectrometer.
Finally, the operating software was operated to subtract the water spectrum from the milk
spectrum. The spectrum of full-cream milk after spectral subtraction of water was
collected. The same procedures were done for the skimmed as well as reduced-fat milk.

Part 2
The sample was inserted into the sample holder and a blank spectrum of the gold plated
surface was recorded. A piece of Si-C abrasive paper was adhered to the surface of the
platen and then placed into the sample holder to obtain the spectrum of the Si-C abrasive
paper. The platen was screwed onto the probe and the Si-C paper was rubbed against the
surface of sample (PMMA) for a few seconds. The platen was unscrewed and placed
onto the sample holder. The spectrum of the sample was obtained. The same procedure
was done on another sample (polystyrene). After recording all the sample spectra, the
operating software was operated to convert the y-axis of the spectrum from reflectance
into absorbance and then substrate the blank from the sample spectra. The OMNIC
software was operated to search the major absorption bands from a library search
functions.

4. Results and Discussion

(1) Comparison of the Milk Contents (ATR)


Four spectra of the milk samples are attached. Three of them are for each of the full-
cream milk, reduced-fat milk, and skim milk, and the other one summarizes all of them
for comparison purpose.

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Because the three milk samples differ in fat content, it is useful to identify the
characteristic peaks that result from fat. In this experiment, the absorption at about 2874
and 1747 cm-1 are used. They correspond to the C-H bond of fatty acid and C=O of ester
linkage.[4]

The absorptions at these two frequencies decrease from full-cream milk, to reduced-
fat milk, and to skim milk. This is in agreement with the fat content in the three milk
samples. It is noted, however, that some absorptions still remain even in the skim milk
sample. This may be due to the residue fat, or may be due to the hydrocarbon and
carbonyl (peptide bond) in the protein.

(2) Identification of Unknown Polymers with DRIFT spectroscopy


The powders ground from a “plastic” tray (Sample 1) and a bucket cover (Sample 2)
were put into the FT-IR spectrophotometer for spectrum acquisition. The spectra were
then compared by the reference spectrum obtained from automated library search.

Spectrum of sample 1 had closest resemblance with that of poly-(methyl


methacrylate), with a similarity of 91.96%. Sample 2 is closest to polystyrene, with a
matching score of 91.85%. The primary source of error is due to a diminished reflectance
observed at lower frequency side. The phenomenon will be discussed later.

poly-(Methyl Methacrylate) poly-Styrene


Diagram 1: Polymer structures

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The major peaks in the spectra are identified and summarized below (in cm-1)

poly-(Methyl Methacrylate) poly-Styrene

Stretching of C-H Stretching of C-H


2900~3000 2820~2920
( Saturated) (Saturated)

Stretching of C-H
1730 Stretching of ester C=O 3000~3100
(Aromatic)

Stretching of C=C
1600
(Aromatic)

Table 1: Identification of the major peaks in the two spectra

(3) Answers to the Questions

1. What are the causes of the prominent absorption features in the background
spectrum around 1200 to 2000 cm-1, 2300 to 2400 cm-1 and 3500 to 4000 cm-1?

Ans. The absorption around 2300 to 2400 cm-1 is due to the asymmetric stretching of
CO2, the absorption around 1200 to 2000 cm-1 is due to the vibrational and rotational
motion of water, the absorption around 3500 to 4000 cm-1 is the overtone of the
absorption peaks caused by water.

2.Why is there a sudden drop off of transmission to zero at frequencies below ~650
cm-1?

Ans. The IR absorption of ZnSe crystal below about 650 cm-1 causes the sudden drop off
of transmission to zero at frequencies below ~650 cm-1.

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3. Can you distinguish the different types of milk by looking at the IR spectrum
alone?

Ans. By just looking at the IR spectrum alone, the types of milk cannot be distinguished,
because the peak caused by IR absorption of fat cannot reflect the fat content in
milk, in fact, the peak of IR spectrum caused by fat is still present in the spectrum of
skim milk (this may caused by the trace amount of residual fat or other components
having the same absorption frequency), it’s impossible to distinguish the type of
milk by just looking at the IR spectrum alone.

4. Why do diffuse reflectance spectra obtain with the Si-C paper scrubbing
technique show a general decrease in reflectance towards the lower frequency end?

Ans. In the diffuse reflectance spectroscopy, the reflected light from the soild sample was
measured. However, in most cases, such light often comes along with specular
reflection, which bounces off the surface instead of penetrating into the depth of the
sample. It can interfere with diffuse reflectance, and cause distortion in the spectrum.
Some papers have described this phenomenon in detail with advanced
knowledges[5-7]. However, for the time being, it might be explained in the following
simple way.

Since diffuse reflection requires


the light to penetrate into the
sample, it inevitably travels
more path than specular
reflection. If the difference in
path length is equal to integer
of wavelength,constructive
Diagram 2: Interference between two kinds of interference occurs. If it is
reflections equal to “integer and half”

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wavelength, destructive interference occurs. So depending on the wavelength, the
reflectance will have slightly (sometimes maybe severely) altered. In the DRIFT
spectrum, the low frequency side is closer to destructive interference, thus the
observed reflectance is smaller.

5. Is background correction a necessity for the identification of the chemical nature


of the sample in diffuse reflectance measurements?

Ans. Background correction is needed for identification of the chemical nature of the
sample in diffuse reflectance measurement. The container used to contain the sample
for the test, e.g. the Si-C paper used in this experiment, would not totally reflect the
incidence light by specular way, it also reflects the light by diffuse reflectance, i.e. it
also absorbs the incidence light as the sample for test. To eliminate the effect of
diffuse reflectance of the container, background correction is needed in order to
make the spectrum of the sample given out more accurate.

5. Conclusion
The ATR infrared spectra of the three types of milk – full-cream, reduced-fat, and
skim – were obtained. The relative fat content was justified from the peaks of spectra.

Also, the compositions of the tray and bucket cover were identified with DRIFT
spectroscopy. They correspond to poly-(Methyl Methacrylate) and polystyrene.

6. References
[1] Skoog D.A, West D.M. Principles of Instrumental Analysis, 5th ed. Saunders,
Philadelphia, 1998 Chapters 16~17

[2] Kincaid J.R, Instrumental Analysis, 2nd ed. 1986 Chapter 8

[3] Smith B.C. Fundamentals of Fourier Transform Infrared Spectroscopy, CRC Press,
Chapter 4

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[4] Fernando A.I, Salvador G., Miguel de la G., Analytica Chemica Acta 513(2004),
401-402

[5] Hembree D.M.(Jr), Smyrl H.R., Journal of Applied Spectroscopy 1989 43(2) 267-274 [6]
Schmitt J.M., Kumar G., Applied Spectroscopy 1996 50(8) 1066-1072

[7] KORTE E.H., STAAT H., Journal of Molecular Structure 1990 218 381-386

7. Appendices
Appendices I~IV are the spectra of three milks (indicated by the title on the upper left
corner) and a graph including all the three spectra for absorption. The correct order are from top
to bottom: Full-cream, reduced-fat, and skim milk. There is a mistake in the original print-out of
this graph.

Appendices V~VI are the spectra for part B, the DRIFT technique for identification of
polymers

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