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) NI Qing (05995048d) MAN Lai Fan (07515458d) Duty Group Lea
der Introduction
YEUNG Shing Hin (07533415d) Theoretical Section LI Lai Ping (07546484d) FUNG Yat
Sing (07524431d) Experimental Section 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 (DRI
FT).........................................................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. Appendice
s...............................................................................
..........................................................................8
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1. Introduction Fourier transform infrared spectroscopy (FTIR) is a measuring te

chnique of getting the infrared spectra by measuring the interferogram (time-dom
ain signal) of the samples with the use of interferometer and then the interfero
gram is Fourier transformed to give the spectrum (frequency-domain functions). F
ourier transform is a data processing tool and infrared spectroscopy is the stud
y of interaction of infrared radiation with matter with the resultant spectrum s
howing the absorption due to various distinctive functional groups, giving a mol
ecular fingerprint of the studied compound. FTIR can be applied for the identifi
cation of the unknown, determination of the quality of the sample and the compon
ents of a mixture. There are mainly two common types of infrared sampling techni
ques-they are the transmission techniques and the reflectance techniques. For th
e transmission mode, the infrared radiation beam passes through the sample direc
tly. 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 polyme
rs, gases, liquids and solids that can be ground into a powder with alkaline met
al halide such as KBr pellet or suspended in an oil. In the reflectance techniqu
e, 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 prop
er amount of light passing through and hence no dilution or grinding of the samp
le is required. And the method is non-destructive. Hence, the technique can be a
pplied to case where it is difficult to study with the transmission technique. T
he attenuated total reflection (ATR) and the Diffuse reflectance infrared Fourie
r transform spectroscopy (DRIFTS) are studied in this experiment. DRIFTS can be
used to examine powder and solid, even very large and intractable samples such a
s pieces of plastic and furniture. ATR can be applied to examine polymer films a
nd 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 reflec
tance 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 refract
ive medium to less refractive one with incident angel greater than critical ange
l. Critical angel, c = sin 1 2 w ere is 1 t e refractive index of more refractive
1 medium and 2 is t e refractive index of less refractive medium. In ATR, a ig l
y refracting prism is used. Alt oug t e IR lig t beam is expected totally refra
cted, evanescent wave will get t roug to t e s allow surface of t e less refrac
tive medium and absorbed by t e sample t at contact to t e prism. B. Diffuse Ref
lectance Infrared Fourier Transform Spectroscopy (DRIFT) Diffuse reflection and
specular reflection are t e components observed of reflection from solid surface
. In DRIFT, lig t s ines onto t e powdered sample, specular component, w ere t e
lig t is reflected directly off, was blocked and diffuse reflectance, w ere t e
lig t penetrated, transmitted and reflected wit in t e sample, was collected an
d analyzed. Bot of t e met ods can use in sample t at is not pulverized or pall
etized easily w ic cause difficulty in analysis by typical met ods.[1
3] 3. Pro
cedure Part 1 (T e power of t e PC ad turned on and t e spectrometer operating
software ad loaded)
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T e parameters suc as resolution were set on t e spectrometer operating softwar

e. We ad made sure t at t e surface of t e ATR prism was cleaned. T e backgroun
d was t en collected by operating t e spectrometer. After t at, a few drops of w
ater were added on t e surface of t e prism by dropper. T e spectrum of water wa
s collected. T e water was removed from t e prism surface by absorbing it away w
it soft tissue paper. T e fullcream milk samples were t en added to t e surface
of prism by t e same steps of adding water. And t e spectrum of full
cream milk
was collected by operating t e spectrometer. Finally, t e operating software wa
s operated to subtract t e water spectrum from t e milk spectrum. T e spectrum o
f full
cream milk after spectral subtraction of water was collected. T e same pr
ocedures were done for t e skimmed as well as reduced
fat milk. Part 2 T e sampl
e was inserted into t e sample older and a blank spectrum of t e gold plated su
rface was recorded. A piece of Si
C abrasive paper was ad ered to t e surface of
t e platen and t en placed into t e sample older to obtain t e spectrum of t e
Si
C abrasive paper. T e platen was screwed onto t e probe and t e Si
C paper w
as rubbed against t e surface of sample (PMMA) for a few seconds. T e platen was
unscrewed and placed onto t e sample older. T e spectrum of t e sample was obt
ained. T e same procedure was done on anot er sample (polystyrene). After record
ing all t e sample spectra, t e operating software was operated to convert t e y

axis of t e spectrum from reflectance into absorbance and t en substrate t e bl
ank from t e sample spectra. T e OMNIC software was operated to searc t e major
absorption bands from a library searc functions. 4. Results and Discussion (1)
Comparison of t e Milk Contents (ATR) Four spectra of t e milk samples are atta
c ed. T ree of t em are for eac of t e fullcream milk, reduced
fat milk, and sk
im milk, and t e ot er one summarizes all of t em for comparison purpose.
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Because t e t ree milk samples differ in fat content, it is useful to identify t

e c aracteristic peaks t at result from fat. In t is experiment, t e absorption
at about 2874 and 1747 cm
1 are used. T ey correspond to t e C
H bond of fatty
acid and C=O of ester linkage.[4] T e absorptions at t ese two freuencies decre
ase from full
cream milk, to reducedfat milk, and to skim milk. T is is in agree
ment wit t e fat content in t e t ree milk samples. It is noted, owever, t at
some absorptions still remain even in t e skim milk sample. T is may be due to t
e residue fat, or may be due to t e ydrocarbon and carbonyl (peptide bond) in
t e protein. (2) Identification of Unknown Polymers wit DRIFT spectroscopy T e
powders ground from a plastic tray (Sample 1) and a bucket cover (Sample 2) were p
ut into t e FT
IR spectrop otometer for spectrum acuisition. T e spectra were t
en compared by t e reference spectrum obtained from automated library searc . S
pectrum of sample 1 ad closest resemblance wit t at of poly
(met yl met acryla
te), wit a similarity of 91.96%. Sample 2 is closest to polystyrene, wit a mat
c ing score of 91.85%. T e primary source of error is due to a diminis ed reflec
tance observed at lower freuency side. T e p enomenon will be discussed later.
poly
(Met yl Met acrylate) Diagram 1: Polymer structures
poly
Styrene
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T e major peaks in t e spectra are identified and summarized below (in cm
1) pol
y
(Met yl Met acrylate) 2900~3000 Stretc ing of C
H ( Saturated) Stretc ing of e
ster C=O poly
Styrene 2820~2920 Stretc ing of C
H (Saturated) Stretc ing of C
H
(Aromatic) Stretc ing of C=C (Aromatic)
1730
3000~3100
1600
Table 1: Identification of t e major peaks in t e two spectra
(3) Answers to t e Questions 1. W at are t e causes of t e prominent absorption
features in t e background spectrum around 1200 to 2000 cm
1, 2300 to 2400 cm
1
and 3500 to 4000 cm
1? Ans. T e absorption around 2300 to 2400 cm
1 is due to t
e asymmetric stretc ing of CO2, t e absorption around 1200 to 2000 cm
1 is due t
o t e vibrational and rotational motion of water, t e absorption around 3500 to
4000 cm
1 is t e overtone of t e absorption peaks caused by water.
2.W y is t ere a sudden drop off of transmission to zero at freuencies below ~6
50 cm
1? Ans. T e IR absorption of ZnSe crystal below about 650 cm
1 causes t e
sudden drop off of transmission to zero at freuencies below ~650 cm
1.
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3. Can you distinguis t e different types of milk by looking at t e IR spectrum

alone? Ans. By just looking at t e IR spectrum alone, t e types of milk cannot
be distinguis ed, because t e peak caused by IR absorption of fat cannot reflect
t e fat content in milk, in fact, t e peak of IR spectrum caused by fat is stil
l present in t e spectrum of skim milk (t is may caused by t e trace amount of r
esidual fat or ot er components aving t e same absorption freuency), its imposs
ible to distinguis t e type of milk by just looking at t e IR spectrum alone.
4. W y do diffuse reflectance spectra obtain wit t e Si
C paper scrubbing tec n
iue s ow a general decrease in reflectance towards t e lower freuency end? Ans
. In t e diffuse reflectance spectroscopy, t e reflected lig t from t e soild sa
mple was measured. However, in most cases, suc lig t often comes along wit spe
cular reflection, w ic bounces off t e surface instead of penetrating into t e
dept of t e sample. It can interfere wit diffuse reflectance, and cause distor
tion in t e spectrum. Some papers ave described t is p enomenon in detail wit
advanced knowledges[5
7]. However, for t e time being, it mig t be explained in
t e following simple way. Since diffuse reflection reuires t e lig t to penetra
te into t e sample, it inevitably travels more pat t an specular reflection. If
t e difference in pat lengt is eual to integer of wavelengt ,constructive Di
agram 2: Interference between two kinds of reflections interference occurs. If i
t is eual to integer and alf
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wavelengt , destructive interference occurs. So depending on t e wavelengt , t e

reflectance will ave slig tly (sometimes maybe severely) altered. In t e DRIFT
spectrum, t e low freuency side is closer to destructive interference, t us t
e observed reflectance is smaller.
5. Is background correction a necessity for t e identification of t e c emical n
ature of t e sample in diffuse reflectance measurements? Ans. Background correct
ion is needed for identification of t e c emical nature of t e sample in diffuse
reflectance measurement. T e container used to contain t e sample for t e test,
e.g. t e Si
C paper used in t is experiment, would not totally reflect t e inci
dence lig t by specular way, it also reflects t e lig t by diffuse reflectance,
i.e. it also absorbs t e incidence lig t as t e sample for test. To eliminate t
e effect of diffuse reflectance of t e container, background correction is neede
d in order to make t e spectrum of t e sample given out more accurate.
5. Conclusion
T e ATR infrared spectra of t e t ree types of milk full
cream, reduced
fat, and
skim were obtained. T e relative fat content was justified from t e peaks of sp
ectra. Also, t e compositions of t e tray and bucket cover were identified wit
DRIFT spectroscopy. T ey correspond to poly
(Met yl Met acrylate) and polystyren
e.
6. References
[1] Skoog D.A, West D.M. Principles of Instrumental Analysis, 5t ed. Saunders,
P iladelp ia, 1998 C apters 16~17 [2] Kincaid J.R, Instrumental Analysis, 2nd ed
. 1986 C apter 8 [3] Smit B.C. Fundamentals of Fourier Transform Infrared Spect
roscopy, CRC Press, C apter 4
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[4] Fernando A.I, Salvador G., Miguel de la G., Analytica C emica Acta 513(2004)
, 401
402
[5] Hembree D.M.(Jr), Smyrl H.R., Journal of Applied Spectroscopy 1989 43(2) 267

274 [6] Sc mitt 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 t e spectra of t ree milks (indicated by t e title on t e up
per left corner) and a grap including all t e t ree spectra for absorption. T e
correct order are from top to bottom: Full
cream, reduced
fat, and skim milk. T
ere is a mistake in t e original print
out of t is grap . Appendices V~VI are t
e spectra for part B, t e DRIFT tec niue for identification of polymers
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