FACULTY OF APPLIED SCIENCES

SPECTROCHEMICAL METHODS OF ANALYSIS

(CHM 580)

EXPERIMENT 1 & 7: FOURIER TRANSFORM INFRARED SPECTROSCOPY (FTIR)

AND NUCLEAR MAGNETIC RESONANCE (NMR) – ANALYSIS OF ASPIRIN-
PHENACETIN-CAFFEINE (APC) TABLET

NAME : HUSNA INSYIRAH BT SAMAD

STUDENT ID : 2017411714
COURSE CODE : AS202
GROUP MEMBERS : NOR AINA ATHIRAH BT MOHD ROZI
SARAH DAYANA BT SHAHRUL BAHARI
LECTURER’S NAME : PN HALIZA BT KASSIM
DATE OF EXPERIMENT: /3/2019
DATE OF SUBMISSION : 23/5/2019
ABSTRACT
In this experiment, each component of APC tablet will be analysed using FTIR and
NMR Spectroscopy. Instead of identifying the functional groups present in standard
compounds Aspirin, Phenacetin and Caffeine, the functional groups of unknown Sample E
also being identified using FTIR Spectroscopy. We use NMR Spectroscopy to identify the
major peaks in NMR spectra of standard compounds of aspirin, Phenacetin and Caffeine. By
having the major peaks of the standard compounds, we can identify which standard
compounds are present in the unknown sample. Hence, we can predict the chemical structure
of unknown sample E by using both data from FTIR and NMR techniques. We prepare the
solid samples of standard compounds and sample E by mixing the samples with KBr. The
functional groups of standard Phenacetin that we identified from the IR spectra are aromatic
and ether. Theoretically, phenacetin should have functional group of aromatic, ether and
amide. In standard Aspirin and ASA, we obtained two functional groups which are
carboxylic acid and aromatic. Functional groups of amine, imine and alkenes are obtained in
standard caffeine. Then, only carboxylic acid group is obtained in sample In NMR spectra,
Phenacetin produces peaks for structure carbonyl, amine, benzene ring , ethoxy and methoxy.
Aspirin produces peak for structure carbonyl, carboxylic acid and benzene ring. ASA only
produces peak for carbonyl and benzene ring. Caffeine produces peak for amine and imine
while sample E produces peak for methyl and methylene structure.

OBJECTIVES
1. To identify functional groups in IR spectra of standard compounds Aspirin,
Phenacetin and Caffeine.
2. To identify functional groups present in aspirin-phenacetin-caffeine tablet.
3. To identify functional groups present in an unknown sample.
4. To identify major peaks in NMR spectra of standard compounds of Aspirin,
Phenacetin and Caffeine.
5. To identify the presence of Aspirin, Phenacetin and Caffeine in unknown tablet
sample.
6. To predict the chemical structure of unknown sample by using both data from FTIR
and NMR technique.

LIST OF CHEMICALS USED

 1. Aspirin
2. Phenacetin
3. Caffeine
4. Aspirin-phenacetin-caffeine tablet
5. KBr pellets of KBr powder
6. CaCl3
7. Acetone

LIST OF APPARATUS USED

1. Agate mortar and pestle
2. Spatula
3. A KBr handpress with a die set
4. Pastuer pipette
5. Conical vial
6. NMR tube

INSTRUMENT
1. PERKIN ELMER SPECTRUM ONE FOURIER TRANFORM INFRARED (FTIR)
2.

EXPERIMENTAL PROCEDURE
Procedure for solid samples:
A. Mixing of sample and KBr:
1. The agate mortar and pestle were removed from the desiccator in the fume hood.
2. 1mg of solid sample was grinded into powder in agate mortar for about 1 minute.
3. 80mg of KBr powder was added into the sample powder and were grinded for about
30 seconds with pestle.
4. The mixture was scraped into the middle with a spatula and the mixture was grinded
again for about 15 seconds.
5. The mixture was heaped in the centre of the mortar by using the spatula.
6. The KBr must kept back into the desiccator after we used.

B. Preparation KBr pellets

1. One fourth of the KBr mixture was took and transferred into the collar of the
handpress.
2. The envil was placed along with die pin so that it came contact with the samples.
3. The de set was lifted carefully by holding the lower envil. The collar must be stayed
in place.
4. The handle of the handpress was opened until the upper ram of the handpress. The
handle was closed.
5. The dial pressure was rotated until the ram of the handpress slightly touched the upper
envil on the die assembles.
6. The unit was tilted back in order to hold the die set from falling off. The handle was
opened.
7. The pressure dial was rotated clockwise in one half turn. The mixture was compress
slowly while closing the handle in 2 minutes.
8. The unit was tilted back, the handle was opened and the die set was removed from the
unit carefully.
9. The pellet was weighed and inspected. The collar containing the KBr pellet was
placed onto a sample holder.

C. Obtaining an IR spectrum
1. There must be nothing in the sample compartment of the IR spectrometer.
2. The instruction provided by instructor was used to operate the spectrophotometer.
3. A background spectrum was obtained.
4. The sample holder was placed in the sample compartment and the data acquisition
was started.
5. If spectrum was not acceptable, a new sample was prepared and the was repeated until
an acceptable spectrum was obtained.

D. Determination of the spectrum of each separate components using NMR

spectroscopy.
1. 30mg of aspirin, phenacetin and caffeine were weighed in different conical vial.
2. About 0.5ml of the deuterated chloroform, CDCl 3 was transferred into the sample
using a clean, dry Pasteur pipette. The conical vial was swirled to help dissolve the
sample.
 3. The sample was completely dissolved after the conical vial was swirled. A little more
solvent was added if necessary to fully dissolve the sample.
4. The solution was transferred into NMR tube using a clean, dry Pasteur pipet. The
solution was transferred carefully to avoid breaking the edge of the fragile NMR tube.
5. A clean pipet was used to add enough deuterated chloroform to bring the total
solution height to about 4-cm from the bottom once the solution has been transferred
to the NMR tube.
6. The NMR tube was capped and make sure the cap was on straight and tight. The
NMR tube was inverted several times to mix the contents.
7. The sample was ready to recorf its NMR spectrum. The NMR tube was inserted into
its holder and adjusted the depth by using the gauge provided.
8. For cleaning purpose, the sample was transferred into same conical vial, partially
refilled with acetone using Pasteur pipet, the cap was replaced carefully and the tube
was inverted several times to rinse it.
9. The acetone was removed and ot was repeated for 2 times and the NMR glass tube
was put into an oven for approximately 2 hours.

E. Analysis of the APC tablet

1. 100mg of the tablet was weighed approximately. The sample was prepared as
described as in Part D.

RESULT
Standard Phenacetin
Vibration Literature wavenumber (cm- Experimental wavenumber
1) (cm-1)
C-H stretching (aromatic) 3100-3000 3073.05
C=C stretching (aromatic) 1600-1430 1447.85
Overtone and combination 2000-1700 2019.59
bands (aromatic)
In-plane C-H bonding 1275-1000 1020.17
(aromatic)
C-O-C stretching (ether) 1100 1106.94

Structure Chemical shift Multiplicity Integral

Literature Experimental
O=CCH 2–3 2.02 – 2.49 Singlet 3H
 -NH 1 -2 1.28 Singlet 1H
Phenyl 6–8 6.83 – 7.45 Doublet 4H
O-CH2 3–4 3.98 Singlet 4H
O-CH2CH3 3-4 3.36 Singlet 3H

Standard Aspirin
Vibration Literature wavenumber (cm-1) Experimental wavenumber
(cm-1)
O-H stretching 3300-2500 2587.67
(carboxylic acid)
C=C stretching (aromatic) 1600-1430 1458.28
Overtone and combination 2000-1700 1754.35
bands (aromatic)
In-plane C-H bonding 1275-1000 1013.35
(aromatic)

Structure Chemical shift Multiplicity Integral

Literature Experimental
O=CCH3 2–3 2.26 – 2.51 Doublet 4H
O=COH 10 -13 13.16 Singlet 1H
Phenyl 6–8 7.22 – 7.94 Quartet 4H

Standard ASA
Vibration Literature wavenumber (cm-1) Experimental wavenumber
(cm-1)
O-H stretching 3300-2500 2546.88
(carboxylic acid)
C=C stretching 1600-1430 1458.28
Overtone and combination 2000-1700 1754.81
bands (aromatic)
In-plane C-H bonding 1275-1000 1013.23
(aromatic)

Structure Chemical shift Multiplicity Integral

Literature Experimental
O=CCH3 2–3 2.28 – 2.51 Doublet 3H
Phenyl 6–8 7.20 – 7.94 Quartet 4H

Standard Caffeine
Vibration Literature wavenumber (cm- Experimental wavenumber
 1) (cm-1)
C-N stretching (amine) 1380–1000 3073.05
C=N stretching (imine) 1690-1640 1660.10
=C-H (alkene) 3100-3000 3113.23
C=C stretching (alkene) 1680-1600 1020.17
=C-H out of plane bending 1000-600 610.16
(alkene)

Structure Chemical shift Multiplicity Integral

Literature Experimental
CH3-N-(C=O)2 2–3 2.49 Singlet 3H
-R-N-CH3 2.3 – 3.0 3.19 – 3.85 Singlet 6H
N=CHN 7.5 – 8.3 8.05 Singlet 1H

Sample E
Vibration Literature wavenumber (cm-1) Experimental wavenumber
(cm-1)
O-H stretching 3300–2500 2675.80
(carboxylic acid)
C=O stretching 1700 1704.17
(carboxylic acid)
C-O-H in-plane bending 1430 1431.47
(carboxylic acid)

Chemical shift Splitting Integration Probable structure

0.85 Triplet 3H R-CH3
1.23 Triplet 2H R-CH2-R

DISCUSSION

Aspirin-Phenacetin-Caffeine is pain relieving tablets that contain aspirin, phenacetin and

caffeine that are represented on the market by various brand names. The components of APC
tablets can be analysed using various techniques. In this experiment, each component of APC
tablet will be analysed using FTIR and NMR Spectroscopy. For standard Phenacetin, the
functional group present in the IR spectra are aromatic and ether. Theoretically, there is
another one functional group which is amide at 3500-3800 wavenumber. In NMR spectra,
Phenacetin produces peak at 2.02 and 2.49 with singlet multiplicity and integral of 3H which
indicates a carbonyl structure. Amine structure was obtained at peak of 1.28 with 1H integral
of singlet multiplicity. Benzene ring was obtained at peak 6.83and 7.45 with 4H integral of
doublet multiplicity. Ethoxy and methoxy structure were obtained at peak of 3.36 and 3.98
with 4H and 3H integral of singlet multiplicity. Aspirin produces peak at 2.26 and 2.51 with
doublet multiplicity and integral of 4H which indicates carbonyl structure. Carboxylic acid
was obtained at peak of 13.16 with 1H integral of singlet multiplicity. Benzene ring was
detected at peak of 7.22 and 7.94 with 4H integral of quartet multiplicity. Supposedly, ASA
should have the same structure as aspirin. But, ASA only produces peak for carbonyl and
benzene ring structure at the same integral and multiplicity as Aspirin. For Caffeine, the
spectrum produces peak at 8.05 that have 1H integral of singlet multiplicity which indicates
imine structure. Amine structure was obtained as there are peak at 3.19 and 3.85 for 6H
integral of singlet multiplicity. In sample E, chemical shift of 0.85 was obtained for methyl
structure with 3H integral of triplet splitting. Chemical shift at 1.23 indicates the methylene
structure with 2H of triplet splitting.

QUESTIONS
1. What is the relationship between wavelength and wavenumber?
Wavelength is inversely proportional to the wavenumber. Wavelength is the distance
over which a cycle repeats while wavenumber is the number of full cycles in a unit
distance.

2. Describe the conditions for IR absorption to occur.

There are two conditions in order for IR absorption to occur. First, there must be a
change in the dipole moment of the molecule as a result of a molecular vibration
(rotation). The change in dipole moment allows interaction with the alternating
electrical component of the IR radiation wave.

3. What is the source of IR radiation?

Heat or thermal radiation.

4. KBr pellet is used for the preparation of the solid sample for FTIR spectrometry.

a) What is the advantage and disadvantage of using KBr pellet.

 The advantage is that KBr has no absorption in the IR above 250cm -1 so that an
unimpeded spectrum of the compound is obtained. The disadvantage is that Br
from the KBr can often replace ligands in the compound whose spectrum is
desired. If this is not realized by the experimenter, misinterpretation of the
spectrum will be resulted.

b) What are the factors caused the unsatisfactory pellet.

Overloading the KBr pellet with the samples causes the pellet to become opaque.
Then, the solid sample is not extremely finely divided and well mixed.

5. We have to calibrate IR instrument before running the sample.

a) What is the purpose of instrument calibration.
To ensure correct response of the instrument.

b) Why do we use polystyrene for calibration.

Polystyrene film is made by the polymer of styrene. Its having highly durability
and stabled at any temperature because it is made by the polymers. The film
produces sharp absorbance bands at known wavelengths which can be used to
verify that the instrument’s spectrum is accurate.

6. Explain how the NMR instrument operates.

The NMR spectrometer generate a constant external magnetic field onto the test
sample, and impose a radio frequency energy on it, via an electric coil to detect the
frequency of the nucleus spin precession (measured the electric signals of the
magnetic field, and by FFT, one can get the spectrum). And the molecular structure
can be determined.

7. What is the chemical shift and what the chemical shift indicates?
Chemical shift is the resonant frequency of a nucleus relative to a standard in a
magnetic field. The position and number of chemical shifts are diagnostic of the
structure of a molecule. Chemical shifts also used to describe signals in other forms of
spectroscopy such as photoemission spectroscopy.

8. What are the factors that affect the chemical shift value?
 Electron density, electronegativity of neighbouring groups and anisotropic induced
magnetic field effects.

9. In NMR spectroscopy, what are the advantages using a magnet with as great as field
strength as possible?
Produces high resolution NMR because resolution directly depends on magnetic field
strength. High resolution NMR does not produces broader peaks which can easily
overlap one another causing issues in resolving complex structures.

10. Why the deuterated solvent is used to dissolve the sample for NMR analysis?
To avoid swamping by the solvent signal, to stabilize the magnetic field and to
accurately define 0ppm.

11. Calculate the chemical shift in ppm for a proton that has resonance 128Hz downfield
from TMS on a spectrometer that operates at 60 Mhz.
δ = ѴH - ѴTMS
ѴNMR
= 128Hz – 0Hz x 106 ppm
60 x 106 Hz
= 2.13 ppm

CONCLUSION
The functional groups in IR spectra of standard compounds Aspirin, Phenacetin and
Caffeine are identified. The functional groups present in the unknown sample E are also
identified. The functional groups of standard Phenacetin that we identified from the IR
spectra are aromatic and ether. In standard Aspirin and ASA, we obtained two functional
groups which are carboxylic acid and aromatic. Functional groups of amine, imine and
alkenes are obtained in standard caffeine. Then, only carboxylic acid group is obtained in
sample E. The major peaks in NMR spectra of standard compound of Aspirin, Phenacetin and
Caffeine are identified. The presence of Aspirin, Phenacetin and Caffeine in unknown tablet
sample are identified. The chemical structure of unknown sample E was predicted by using
both data from FTIR and NMR techniques which Palmitic acid.

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