SPECTROSCOPIC AND CHROMATOGRAPHIC METHODS

PRACTICUM OFFICIAL REPORT

INFRARED SPECTROPHOTOMETER (FTIR)

Compiled by:

Mawar Khurrotul Khasaniyah / 23030234089 / KA 2023

CHEMISTRY STUDY PROGRAM

DEPARTMENT OF CHEMISTRY
FACULTY OF MATHEMATICS AND NATURAL SCIENCE
STATE UNIVERSITY OF SURABAYA
2025
A. Experiment Title
InfraRed Spectrophotometer (FTIR)

B. Experiment Purpose
1. Making the infrared spectrum of a compound
2. Interpreting the infrared spectrum of a compound

C. Experiment Date
Wednesday, 23th April 2025

D. Experiment Time
01.00 – 03.30 pm

E. Literature Review
1. FTIR
One instrument that uses the principle of spectroscopy is
FTIR (Fourier Transform Infrared). Fourier Transform Infrared is
infrared spectroscopy equipped with a fourier transform for
detection and analysis of the spectrum results. Infrared spectroscopy
has benefits for the identification of organic compounds because of
its very complex spectrum, a complex spectrum because it consists
of many peaks that indicate the presence of functional groups
characterized by wave numbers. The purpose of IR analysis is to
estimate the functional groups contained in the complex compound
and determine the shift in functional groups from the initial ligand
to the complex compound formed (Sanjiwani et al., 2020).
This FTIR instrument is used to analyze organic and
inorganic molecules qualitatively and quantitatively with a wave
number range of 14000 cm-1-10 cm-(Faturachman et al., 2025). The
FTIR method has several advantages including simple preparation,
fast scanning, high resolution, does not take a long time, does not
damage, almost all compounds show absorption of infrared radiation
 and without using harmful reagents (Auha & Alauhdin, 2021).
Meanwhile, the disadvantage of this method is that the sampling
space is relatively small, which may block the infrared rays. The
validation parameters in this method are precision, accuracy,
linearity, limit of detection (LOD), and limit of quantification (LOQ)
(Faturachman et al., 2025).
In qualitative analysis, it is used to determine the structure
and functional groups of the compounds being analyzed. Analysis
of the functional group of a compound is done by comparing the
absorbance band formed in the infrared spectrum using the spectrum
of the comparison compound. Meanwhile, quantitative analysis is
used to determine the concentration of a compound from the
analyzed sample. The principle in quantitative analysis is that the
energy absorbed at a certain wave number is proportional to the
number of energy-related bonds, so with a greater concentration of
compounds (Faturachman et al., 2025).

2. FTIR Working Principle

FTIR spectroscopy or infrared spectroscopy is an analytical
method based on the principle of interaction of a chemical
compound with electromagnetic radiation which will produce a
vibration (vibration) of a polyatomic chemical bond or functional
group of chemical compounds (Moros et al., 2010). The working
principle of FTIR is the interaction between energy and material.
Some infrared passing through the slit to the sample is absorbed by
the sample and others are transmitted through the sample surface so
that infrared light passes through the detector and the measured
signal is then sent to the computer and recorded in the form of peaks
(Faturachman et al., 2025).
 Figure 1. Infrared Spectroscopy Schematic (Auha &
Alauhdin, 2021)
The technique of reading molecular vibrations in IR
spectrometry can be used in two variants, namely transmission and
reflectance. The transmission method is used to test the effects of
the absorption of IR radiation in the sample volume. It is possible to
test samples in solid, liquid and gaseous form using the right
procedure. In this method, the sample is directly exposed to laser
light to determine the vibrational pattern of the molecules. In this
method, solid samples are made into clear pellets with the help of
KBr. KBr sample pellets are then analyzed using an FT-IR
spectrophotometer. Measurements were made in the range of wave
numbers / cm-1 4000-400. reflectance measurement using UATR is
one of the techniques in measuring molecular absorption / vibrations
with an FT-IR spectrophotometer. The sample is not directly
exposed to laser light, but is reflected or reflected to determine the
vibrational pattern of the molecule (Sulistyani & Huda, 2018).
As is well known, the atoms in a molecule do not stand still
but vibrate. The energy of most molecular vibrations corresponds to
the infrared region. Molecular vibrations can be detected and
measured in the infrared spectrum. When infrared radiation is passed
through a snippet, the molecules can absorb (absorb) energy and
there is a transition between the ground state and excited state
vibrational levels. The absorption of energy at various frequencies
can be detected by an infrared spectrometer, which plots the amount
of infrared radiation transmitted through a sample as a function of
 the frequency (or wavelength) of the radiation. The plot is called an
infrared spectrum which will provide important information about
the functional groups of a molecule (Sulistyani & Huda, 2018).

3. Function Group Spectrum

Infrared spectroscopy has benefits for the identification of
organic compounds because of its very complex vibrational
spectrum, a complex spectrum because it consists of many peaks that
indicate the presence of functional groups characterized by wave
numbers. The IR spectrum is the amount of IR radiation that is
transmitted through a snippet as a function of the frequency
(wavelength) of the radiation (Sanjiwani et al., 2020).

Table 1.1 Table of IR groups (Sanjiwani et al., 2020)

In the infrared spectrum, the wave number is plotted on the

X-axis and the percent transmittance (%T) is plotted on the Y-axis
(Figure 2). The wave number, sometimes called the frequency, is
proportional to the radiant energy. Percent transmittance can also be
expressed as absorbance, which is the intensity of absorption by the
sample molecules (Auha & Alauhdin, 2021). The relationship
between absorbance and transmittance is expressed by the following
equation:
 Gambar 2. Examples of infrared spectra, (a) Wave number
vs percent transmittance (%T), (b) Wave number vs percent
absorbance (A) (Auha & Alauhdin, 2021)
Infrared spectra containing absorbance or percent
transmittance relationships with wave numbers provide information
about the structure of a molecule. The region at wave numbers
4,000-1,500 cm-1 provides information about the main functional
groups, while the 1,500-1,000 cm-1 region is called the fingerprint
region (Auha & Alauhdin, 2021).
F. Tools and Materials
a) Tools
1. PerkinElmer Spectrum Two Infra Red Spectrophotometer
2. Beaker glass 100 mL 2 pieces
3. Spatula - 1 piece

b) Materials
1. Compound X

G. Experiment Flow
1. Operation Phase
a. Basic operations

Instrument IR
1. Connect the FTIR cable to the socket
2. Click the instrument buttom
3. Turn on the computer
4. Open the software
IR Spectrum software display

5. Make sure the scan, background, and monitor menus

6. Clean the diamond set and sample holder with alcohol
7. Click background and wait until 100%
Liquid sample

8. Fill in the sample 10 according to the name of the

sample being background
9. Pour directly into the diamond circle
10. Install the iron tube on the lever
11. Point the lever at the center of the diamond cell circle
12. Click the sample table, make sure the preview menu is
ready checked
13. Click scan, then turn the level clockwise until pressure
reaches 60 on the scale
14. Click scan again, wait until 100% for the peak to appear
15. Lift and take a sample on the diamond
16. Clean with alcohol
Result (spectrum)
b. Printing spectrum

Spectrum
1. Right click then add text to add text
2. Click file and select a storage location
3. Click the lable menu then print and save according to
the name
4. Click process, click peak, then print and save (for bring
up the table)
5. Click print then select 5. Click print then select
storage and save location storage and save location

PDP Excel/CSV

c. Split the spectrum

Data in CSV format
1. Click split on the menu bar
2. Select the split type
3. Right click then add text (to add information on each
spectrum)
4. Click print file, select a storage location the save

Result

d. Shows peaks in more detail

Peak
1. Click set up
2. Click peak detection
3. In the sholds menu, reduce the peak level from 0,0% to 0,5%
4. Click print to refresh
5. Click back on the label menu
6. Turn off labels menu
Result

e. Turning off the instrument

Instrumen IR
1. Clean the diamond set and sample holder with alcohol
2. Close the IR spectrum software
3. Remove all cables
4. Turn off the computer
IR Spectrum
2. Sample Testing

Sample compoung X
1. Connect the FTIR cable to the socket
2. Click the instrument buttom
3. Turn on the computer
4. Open the software
5. Make sure the scan, background, and monitor menus are
active
6. Clean the diamond set and sample holder with alcohol
7. Click background and wait until 100%
8. Fill in the sample 10 according to the name of the
sample being analyzed
9. Pour directly into the diamond circle
10. Install the iron tube on the lever
11. Point the lever at the center of the diamond cell circle
12. Click the sample table, make sure the preview menu is
ready checked
13. Click scan, then turn the level clockwise until pressure
reaches 60 on the scale
14. Click scan again, wait until 100% for the peak to appear
15. Lift and shake a sample on the diamond set and clean
with alcohol.
16. Add text, right click then add text
17. Save the resulting spectrum by clicking file, selecting a
location and save.
IR spectrum

3. Interperting the IT spectrum

Instrument IR
1. Determine the presence or absence of a number of functional
groups
2. Information on the structure the compound at the peak is
given C=O, O–H, N–H, C–O, NO2
3. The mine check is carried out on functional groups according
to the sample as measured by its
IR Spectrum
H . Observation Results

No Experimental Procedure Observation Results Reaction Conclusion

Before After
−1
1. Operation Phase Sample X Sample X + Based on the practicum
3320 − 3310 𝑐𝑚
(s): White Methanol: : Alkyne C-H that has been done, it
a. Basic Operation powder Colorless Stretch can be concluded that
solution −1 sample x is form nine
Methanol: 1760 − 1740 𝑐𝑚 peak in analysis FTIR
Colorless Sample X + : Alkyl carbonate
−1 3317.98: Alkaline C-H
FTIR: Form 1490 − 1410 𝑐𝑚 stretch
nine peak : Carbonate ion 2943.41: Alkana
−1
1225 − 950 𝑐𝑚 : 2831.39: O-CH₃, C-H
Liquid 1741.71: Alkyl
Aromatic C-H
sample: −1 Carbonate
−1 1100 − 900 𝑐𝑚 : 1449.42: Carbonate Ion
3317. 98 𝑐𝑚 Silicate ion
−1 1214.53: Aromatic C-H
−1
2943. 41 𝑐𝑚
−1 705 − 570 𝑐𝑚 : 1114.28: Aromatic C-H
2831. 39 𝑐𝑚 C-S Stretch 1020.63: Silicate ion
−1
1741. 71 𝑐𝑚 (Nandiyanto et.al., 625.73: C-S Stretch
−1 2019)
1449. 42 𝑐𝑚 −1
Possible compounds
−1 2853 − 2962 𝑐𝑚
1214. 53 𝑐𝑚 : Alkana carbonate ester
−1
1114. 28 𝑐𝑚 (Sanjiwani et.al.,
−1
1020. 63 𝑐𝑚 2020)
−1 −1
625. 73 𝑐𝑚 2850 − 2815 𝑐𝑚
: Methyl ether
O-CH₃, C-H stretch
(Nandiyanto et.al.,
 2019)

B. Printing Spectrum Solid sample Solid sample

−1 −1
3413. 72 𝑐𝑚 3413. 72 𝑐𝑚 : H
−1
1664. 55 𝑐𝑚 Bonding
−1
−1
1112. 36 𝑐𝑚 1664. 55 𝑐𝑚 :
C=C Alkene
−1
1112. 36 𝑐𝑚 :
C-Namine

C. Split the Spectrum

D. Shows Peaks in More Detail

E. Turning off The Instrument

2. Sample Testing

3. Interpreting The IT Spectrum

I. Analysis
The practicum entitled Infrared Spectrophotometer (FTIR) was
carried out on Wednesday, April 23, 2025, at 01.00 - 03.30 WIB in the
Analytical laboratory of Surabaya State University. This practicum aims to
(1) Making the Infrared spectrum of a compound, (2) Interpreting the
infrared spectrum of a compound.
This practicum was conducted using an InfraRed spectophotometer.
There is a difference between spectophotometry and spectophotometer.
Spectophotometry is the method of analysis used, while the
spectophotometer is the tool used. Spectophotometry is an analytical
method based on the absorption of photons, which are particles of light that
cause electromagnetic radiation. Electromagnetic radiation is an
electromagnetic wave that has a spectrum with a certain wavelength.
Electromagnetic radiation consists of various kinds, namely radio waves,
microwave waves, infrared waves, and color waves. Color Spectrum
Waves., X-Ray Waves, and Gamma Ray Waves.
The spectrophotometer used in this experiment is an IR
spectrophotometer. IR spectophotometer is used to determine the functional
groups or substituents of a compound based on the absorption pattern of
infrared light by molecular bonds. Functional groups are the constituent
substituents of a compound. IR spectrophotometer can identify functional
groups because each type of functional group in the molecule has a
characteristic vibrational frequency that can absorb infrared light at a certain
wavelength, resulting in a unique spectrum pattern (fingerprint) or produce
peaks that can be used to recognize the presence of these functional groups
in a compound. In addition to vibrational energy, various kinds of energy
are known in physics, such as translational energy, rotation, etc. Vibrational
energy itself is vibrational energy while this peak is the area with the highest
or optimal energy.
Each compound that has a similar or same functional group, the
absorption area will be typical. If a compound has the same functional
group, then in an IR instrument they will show an absorption band (peak) in
almost the same region of the spectrum, because the functional group
absorbs infrared radiation at a similar vibrational frequency. For example, a
compound has an OH functional group, when it is fired by IR radiation that
absorbs a certain radiation, it is the functional group.
In the experiment conducted, sample X was obtained in liquid form
in the form of a colorless solution. In addition to liquid samples, there are
also samples in the form of solids in the form of white powder which are
tested by other groups. The difference is that for liquid samples, they are
diluted first into a solution using methanol. The use of methanol as a solvent
aims to produce more accurate analysis results because methanol is cleaner
and not easily contaminated by impurities that can affect the analysis results.
The steps in this experiment can be divided into the following:
1. Sample Preparation
Sample preparation aims to prepare samples before being tested using
IR spectophotometry. For solid samples, the first step is to weigh the
sample x as much as 1 gram and then put it in a small beaker. For liquid
samples, after weighing 0.3 grams using an analytical balance, the next
step is to dissolve it in a 10 ml volumetric flask with methanol solvent.
The step is to dissolve sample x first in a beaker and then put methanol
and then stirred and then put it in a 10 ml measuring flask.
2. Operation Stage
This stage contains the steps in running IR spectrophotometry
correctly starting from the process of turning on to printing the
spectrum. This stage is divided into several stages, namely:
a. Operation stage
This stage aims to find out how to operate IR spectrophotometry
starting from the process of turning on until the spectrum is obtained.
The first step in this experiment is to connect the FTIR cable to the
socket first. After that click the instrument button and get a computer
that turns on. After the computer turns on. The next step is to open
the IR spectrophotometer software. After the application is open, the
next step is to make sure the soon, background and monitor menus
are active. Once active, you can then clean the diamond set and
sample holder using methanol. The purpose of using methanol to
clean the diamond set is to clean the set from contaminants without
affecting and interfering with the analysis process because it does
not absorb waves and methanol only cleans the diamond set because
of its volatile nature. After the diamond set has been cleaned, the
baground is clicked and waited until 100%. The next step is to fill in
the sample ID with the name of the sample being analyzed. Next,
sample x is placed on the diamond set circle until it covers the black
circle on the diamond set. Once covered, the iron tube is rotated on
the lever to be directed right in the center of the diamond set circle
for solid samples. However, for diamond set circle samples that have
been dripped with samples, there is no need to rotate the iron tube
on the lever. After closed fitting in the middle of the sample then
click scan, and rotate clockwise until the pressure reaches scale 60.
Then scan back to 100% to bring up the peak. After the peak appears,
the next step is to remove the sample from the diamond set and clean
it again using methanol and tissue.The result of this step is that the
results are obtained in the form of a spectrum. The following is the
spectrum produced by our group from sample x :
 b. Printing the Spectrum
This step aims to find out how to save the spectrum results.
The first step, right-click on the spectrum then add text to add text.
Next, select the storage location and then save. Click the label menu,
then print and save according to the desired name to display the label
on the silver. Click process then select peak table then save. To save
it can be in pdf or Excel / CSC. To save in pdf form, just do it directly
like the initial step and to save it in excel form, click export then
select save.
c. Squinting the spectrum
The first step is that the data in CSV form is clicked on the
menu bar, then select the split type then right-click then add text.
Next, just print the file and then select the storage location for the
file.
d. Turning off the instrument
This step aims to find out how to turn off the IR instrument
correctly. First, clean the diamond set using methanol, then close the
IR spectrum software. Next, turn off the computer as usual and don't
forget to remove all labels.
3. Interpreting IR spectra
The first step in this process is to determine the presence or absence
of a number of functional groups. The trick is to find the IR absorption
theory for each cluster in the IR cluster table. If so, then list the wave
numbers that appear on the peak and then analyze which part of the peak
is included in the functional group that has been made the theory. After
that, the main check of the functional groups of the chemical structure is
carried out.
 From the results of the spectrum produced by our group using liquid
samples, a total of 9 peaks were obtained, from these 9 peaks can be divided
into stretching and bending vibrations. Stretching in the peak spectrum is a
peak that has a loose-shaped spectrum and is not too sharp, while for binding
is a peak that is close together, tight and pointed. Based on the theory we
use, namely for each functional group and its absorption area is as follows:
1. 3320-3310 cm-1 = C-H alkyne stretch
2. 1760-1740 cm-1 = Alkyl carbonate
3. 1490-1410 cm-1 = Carbonate ion
4. 1225-950 cm-1 = C-H aromatic
5. 1100-900 cm -1 = Silicate ion
6. 705-570 cm-1 = C-S stretch
7. 2850-2815 cm-1 = metoxy, methyl eter O-CH3, C-H stretch
(Nandiyanto et al., 2019)
8. 2853-2962 cm-1 = Alkana
(Sanjiwani et al., 2020)
Based on the analysis of the absorption area, 9 peaks were obtained
with details, C-H alkyne stretch group with a spectrum of 3317.98 cm-1,
alkane group with a spectrum of 2943.41 cm-1, O-CH3 and C-H groups with
a spectrum of 2831.39 cm-1, alkyl carbonate group with a spectrum of
1741.71 cm-1, carbonate ion group with a spectrum of 1449.42 cm-1,
 aromatic C-H group with a spectrum of 1214.53 and 1114.28 cm-1, silicate
ion group with a spectrum of 1020.63 cm-1, and C-S stretch group with a
spectrum of 625.73 cm-1. While the solid sample produced 3 peaks, namely
the H bonding group at 3413.72 cm-1 spectrum, C=C alkene group at
1664.55 cm-1 spectrum, and C-N amine group at 1112.36 cm-1 spectrum.
The red graph is the result of measurement by group 3 using solid
samples and the black graph is the result of measurement by group 5 using
liquid samples. From the results of the functional groups in sample x testing,
it is likely that compound x is a carbonate ester.

J. Conclusion
Based on the experiments that have been carried out, the conclusions
obtained from this experiment are:
1. The cluster data obtained in the liquid X sample are C-H alkyne stretch
group with a spectrum of 3317.98 cm-1, alkane group with a spectrum
of 2943.41 cm-1, O-CH3 and C-H groups with a spectrum of 2831.39
cm-1, alkyl carbonate group with a spectrum of 1741.71 cm-1, carbonate
ion group with a spectrum of 1449.42 cm-1, aromatic C-H group with a
spectrum of 1214.53 and 1114.28 cm-1, silicate ion group with a
spectrum of 1020.63 cm-1, and C-S stretch group with a spectrum of
625.73 cm-1
2. The difference in cluster data between the liquid X sample and the solid
X sample is obtained, namely the liquid X sample has more peaks than
the solid X sample. In addition, the functional groups detected in the
liquid X sample are more than the solid X sample.

K. Suggestion
The advice that can be given is that when using an infrared
spectrophotometer instrument, you must be careful not to damage the
instrument because it is a fairly expensive tool for now. In addition, in
analyzing using infrared spectrophotometers, it is necessary to first learn to
 know how to operate an infrared spectrophotometer instrument so that the
resulting graph is as desired.

L. References

Auha, A. N., & Alauhdin, M. (2021). Indonesian Journal of Chemical

Science Development and Validation of Infrared Spectroscopy
Methods for Rutin Compound Analysis. Indonesian Journal of
Chemical Science, 10(2). http://journal.unnes.ac.id/sju/index.php/ijcs
Faturachman, G. F., Ramanda, A. A., Maharani, S., Latif, L. A., Belo, G. A.
G., & Ayubi, S. G. Al. (2025). Application of Fourier Transform
Infrared Spectroscopy (FTIR) for Quantitative Analysis of
Pharmaceutical Compounds. Indonesian Journal of Pharmaceutical
Education, 5(1). https://doi.org/10.37311/ijpe.v5i1.23309
Moros, J., Garrigues, S., & de la Guardia, M. (2010). Vibrational
spectroscopy provides a green tool for multi-component analysis. TrAC
Trends in Analytical Chemistry, 29(7), 578–591.
Sanjiwani, N. M. S., Mariati, N. P. A. M., Sudiarsa, I. W., Paramitha, D. A.
I., Chandra, A. A. W., Ariawan, I. M. D., Megawati, F., & Dewi, N. W.
T. (2020). PEMBUATAN HAIR TONIC BERBAHAN DASAR LIDAH
BUAYA DAN ANALISIS DENGAN FOURIER TRANSFORM
INFRARED. 21(1), 249–262.
Sulistyani, M., & Huda, N. (2018). Indonesian Journal of Chemical Science
Perbandingan Metode Transmisi dan Reflektansi pada Pengukuran
Polistirena Menggunakan Instrumentasi Spektroskopi Fourier
Transform Infrared. Indonesian Journal of Chemical Science, 7(2).
http://journal.unnes.ac.id/sju/index.php/ijcs

M. Attachment
a. Question and Answer
1. Can the sample to be analyzed by FTIR contain water?
Explain your answer!
In general, water should be avoided in FTIR samples unless it is
an inherent part of the analyte being studied. This is because
water contains O–H bonds that absorb very strongly in the
infrared region, particularly near 3400 cm⁻¹ (O–H stretching)
and 1640 cm⁻¹ (O–H bending). These absorption bands are
 typically broad and intense, which can interfere with or obscure
the peaks of other functional groups. Consequently, this can lead
to inaccurate identification of compounds and, in some cases,
make quantification unreliable or impossible.

2. From the interpretation of the infrared spectrum of a

compound, can the structure of the compound be known?
structure of the compound? Explain your answer!
In general, FTIR spectral interpretation cannot definitively
or completely determine a compound’s full molecular structure,
but it is highly effective for identifying the functional groups
present in a molecule. FTIR operates on the principle that
chemical bonds between atoms absorb infrared radiation at
specific frequencies, which are influenced by the mass of the
atoms and the strength or stiffness of the bond. Since each
functional group exhibits characteristic vibrational frequencies
(vibrational modes), they can be detected by their unique
absorption peaks in the IR spectrum.

3. Explain whether infrared spectrum data can be used to

know that an organic chemical reaction can take place?
Yes, infrared (IR) spectral data can be effectively used to
monitor or confirm that an organic reaction has occurred,
particularly when it involves changes in functional groups.
Since FTIR detects specific bond vibrations within molecules,
the emergence or disappearance of certain absorption peaks in
the IR spectrum can signal a chemical transformation. For
instance, during the reduction of a carbonyl group (C=O) to an
alcohol (–OH), the distinct peak near 1700 cm⁻¹, characteristic
of the C=O bond, will vanish and be replaced by a broad
absorption around 3200–3600 cm⁻¹, indicating the formation of
a hydroxyl group. Likewise, in an esterification reaction
 between a carboxylic acid and an alcohol, the resulting spectrum
will show the loss of the –OH peaks associated with the starting
materials.

b. Calculation
❖ Known:
➢ Mass: 0.3gr = 300 mg
➢ Diluted in 10 mL = 0.01 L
❖ M: 𝑀𝑎𝑠𝑠 (𝑚𝑔)/ 𝑉𝑜𝑙𝑢𝑚𝑒 (𝑙) = 300 (𝑚𝑔)/ 0.01 𝑙 = 30.000 𝑝pm
c. Documentation

Weighed sample X as Result of weighed Diluted sample X

much as 0,3 grams sample

Result of dilutes sample Result of spectrum IR

in measuring flask sample X
PRALAB
18/4/2025