ANALYSIS BY INFRARED (IR) SPECTOPHOTOMETRY

By Bernabas Bio (1013031017)

Chemistry Education Department, Faculty of Mathematic and Natural Sciences
Ganesha University of education. Udayana Street No. 11 Singaraja Bali 81117 Indonesia

INTRODUCTION
The Infrared (IR) spectrometry is a method of both qualitative and quantitative
analyses which based on the absorption of IR radiation. It has the wavelength region 0.75 -
1000 μm, and the wave number in the range of 13000-10 cm-1. The IR absorption spectrum
provides a “fingerprint” of a molecule with covalent bonds so can be used to identify the
molecule. In addition, the spectrum provides a quick way to check for the presence of a
given functional group in a molecule. The entire IR region can be divided into the near-IR
(from 0.75 to 2.5 mm), the mid-IR (from about 2.5 to 20 mm), and the far-IR (from 20 to
200 mm). (Robinson, et al: 2005). The wavelength used in infrared spectroscopy
equipment is on mid-IR region that is at wavelengths from 2.5 to 50 μm or the wave
number 4000-200 cm-1. The area is suitable for molecular changes in the vibration energy.
The far-IR region (wave number from 400 to10 cm-1), is useful for molecules containing
heavy atoms, such as inorganic compounds but rather requires a specific experimental
technique. But the common used is the IR spectrum which has the wave number in the
range from 4000 to 670 cm-1. This spectrum is related to the vibration and rotation
energies of molecules. (Muderawan: 2009).
Molecules with covalent bonds may absorb IR radiation and this absorption is
quantized, so only certain frequencies of IR radiation are absorbed. When radiation, (i.e.,
energy) is absorbed, the molecule moves to a higher energy state. The energy associated
with IR radiation is sufficient to cause molecules to rotate (if possible) and to vibrate.
Molecules absorb radiation when a bond in the molecule vibrates at the same frequency as
the incident radiant energy. After absorbing radiation, the molecules have more energy and
vibrate at increased amplitude. The frequency absorbed depends on the masses of the
atoms in the bond, the geometry of the molecule, the strength of the bond, and several other
factors. Note that not all molecules can absorb IR radiation. The molecules that only have
a change in dipole moment during vibration in can absorb IR radiation.
The common molecular vibrations that are excited by IR radiation are stretching
vibrations and bending vibrations. The stretching vibration involves a change in bond
lengths resulting in a change in interatomic distance, while the bending vibration involves
a change in bond angle or a change in the position of a group of atoms with respect to the
rest of the molecule. Generally, there are two types of stretching vibration, namely
symmetrical and asymmetrical stretching, and also there are four types of bending
vibration, namely scissoring, rocking, wagging, twisting. The symmetric stretching
vibration does not change the dipole moment of the molecule. This vibrational mode does
not absorb IR radiation so called IR-inactive. However, the other two modes of vibration
do change the dipole moment so called IR-active. The asymmetric stretching frequency
occurs at 2350 cm-1 and the bending vibration occurs at 666 cm-1. These types of
vibrations are drawn as follow:

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 Figure 1. Principal modes of vibration between carbon and hydrogen in an alkane
In accordance with quantum mechanics, energy is quantized, so the vibration
energy of molecules is also quantized. The frequency of vibration of the bond in the
1 h k k
molecule can be predicted by using the equation: v    5,3  10 12
 2 c  

In which v = 1/  is wave number (cm-1), h is Planck's constant, k is bond constant (N/m), c

adalah the speed of light (cm/dt) and  is reduced mass (g).
The most IR spectrometer systems were double-beam dispersive grating
spectrometers, similar in operation to the double-beam system for UV/VIS spectroscopy.
But now, these instruments have been replaced almost entirely by FTIR spectrometers
because of the advantages in speed, signal-to-noise ratio, and precision in determining
spectral frequency that can be obtained from a modern multiplex instrument.
Solid and Liquid samples
In general, spectra from solid samples are used for qualitative identification of the
sample, not for quantitative analysis. The spectrum of a solid sample is generally collected
when the sample is not soluble in a suitable IR-transparent solvent. But there are some
problems can be occurs with spectra from solid samples. Many organic solids are
crystalline materials. The mull and pellet approaches result in random orientation of the
finely ground crystals; deposition of thin films by evaporation may result in a specific
crystal orientation with respect to the light beam. However, liquid samples are the easiest
samples to be handled. Many liquids may be analyzed “neat”, that is, with no sample
preparation. Neat liquids that are not volatile are analyzed by pressing a drop of the liquid
between two flat salt plates to form a very thin film. The salt plates are held together by
capillary action or may be clamped together. NaCl and KBr are used as the plates. Volatile
liquids may be analyzed neat by using a pair of salt plates with a thin spacer in a sealed
cell. The path length of these cells depends on the spacer thickness. For neat liquids very
small path lengths, less than 0.05 mm must be used to avoid complete absorption of the
source beam. Sample sizes used for the collection of neat spectra are on the order of a few
milligrams of material.

METHODS
Equipments, Chemicals, and Instrument
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Equipments:
The equipments used in this experiment are mortar and pestle, spatula, pipette, and beaker
glass (100 mL).
Chemicals:
The chemicals used in this experiment are NaBr, unknown compound: A (solid) and B
(liquid), mixture of elaidate-oleic acid 25%, 50%, and 75%.
Instrument:
Instrument used is FTIR spectrophotometer
Procedures:
Qualitative Analysis
The measurement of the transmitance of KBr plat was done by using FTIR instrument
in the range of vibration 500 – 4000 cm-1. The unknown sample A (solid) and B (liquid)
were prepared and then their transmitance was measured by using FTIR spectrophotometer
in the range of vibration 500– 4000 cm-1. It was gotten the characteristic of transmitance
and then the functional group of these unknown compounds can be known.
Quantitative Analysis
The spectrum of a mixture of oleic-elaedic acids was measured using an infrared
spectrophotometer. The spectrum of unknown percentage of mixture of oleic -elaedic acid
was measured. Measurement of VCO was performed. After the absorbance of oleic-elaedic
acid the concentration can be known. Then the concentration of olaedic-oleic acid and the
concentration of oleic in ester VCO were determined.
EXPERIMENT RESULTS AND DISCUSSION
Infrared spectrometry is the main method in determining the molecular structure of
organic and biochemistry. Therefore, in experiments that used infrared spectrometry
sample most of the organic compounds. According to the table of observations, the
infrared spectrometer experiment was carried out qualitative and quantitative analysis of
organic samples has been prepared. In a qualitative analysis of two samples used is
unknown-solid samples (sample A) and liquid samples (sample B).In the quantitative
analysis was measurement of the spectra of elaedic-oleic acid mixture of 25%, 50%, and
75% as well as measurements of the VCO ester. From the results of measurements of this
spectrum, concentration of elaedic-oleic acid and oleic in ester VCO can be determined.
From IR analysis, the results of unknown liquid and solid samples are are shown as
follow:

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 Figure 1. The measurement result of Unknown sample A (solid)
From the result, so we get data:

Table 1. Unknown solid sample

FUNCTIONAL GROUP
Frequency range
BOND Frequency range COMPOUND
(cm-) -1
(cm )
2970 C-H 2850-2970 Alkanes
1710 C=O 1690-1760 Aldehyde, ketone,
carboxylic acid, ester
1050 C-O 1050-1300 Alcohol, ether, carboxylic
acid, ester
Based on the data, it can be predicted that the compound included to esters group. So, can
be determined the constant of C-O with the following calculation:

Known : wave number  v  = 1050 cm-1=1.050 x 105 m-1, and c = 3 x 108 m/s


 
Asked: k =…….?
Answer:
Mass of carbon:
12 10 -3 kg / mol
m1  1 atom 2.0  10  26 kg
6.02  10 23 atom / mol

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Mass of oxygen:
16 10 -3 kg / mol
m2   1 atom 2.7  10  26 kg
6.02  10 atom / mol
23

  1.15 10 26 kg

k
  5.3 10 -12

k
1050 cm 1  5.3 10 -12 s / cm
1.15 10 26 kg
1050 cm 1 k

5.3 10 s / cm
-12
1.15 10 26 kg
k
198.113  1012 s 1 
1.15 10 26 kg
k  ( 198.113  1012 s 1 ) 2 (1.15 10 26 kg)
k  (39248.843  10 24 s  2 )(1.15 10  26 kg)
k  45136.169  10  2 kg s  2
k  451.361 kg s  2
k  0.451361 10 3 N / m
So, constant bond of C=O in the liquid sample B is 0.451361x 103 N/m

Figure 2. The measurement result of unknown sample B (liquid)

Table 2. Unknown liquid sample

Frequency Bond Frequency Compound Intensity

range range (cm-1)
(cm-)
2910 C-HStr 2850-2970 Alkane Strong
1730 C=O 1690-1760 Aldehyde, ketone, Strong
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 carboxylic acid, ester
1380 C–Hbend 1340-1470 Alkane Strong

Based on the data above, it can be predicted that the compound contain C=O group
(aldehyde group). So, the bond constant of C=O bond can be determined with the
following calculation:
Known : wave number  v  = 1730 cm-1=1.730 x 105 m-1, and c = 3 x 108 m/s


 
Asked: k =…….?
Answer:
Mass of carbon:
12 10 -3 kg / mol
m1  1 atom 2.0  10  26 kg
6.02  10 atom / mol
23

Mass of oxygen:
16 10 -3 kg / mol
m2   1 atom 2.7  10  26 kg
6.02  10 atom / mol
23

  1.15 10 26 kg

k
  5.3 10 -12

k
1730 cm 1  5.3 10 -12 s / cm
1.15 10  26 kg
1730 cm 1 k

5.3 10 s / cm
-12
1.15 10  26 kg
k
326  1012 s 1 
1.15 10  26 kg
k  ( 326  1012 s 1 ) 2 (1.15 10 26 kg)
k  (106276  10 24 s  2 )(1.15 10  26 kg)
k  122217.4  10  2 kg s  2
k  1222.174 kg s  2
k  1.222  10 3 N / m
So, constant bond of C=O in the liquid sample B is 1.222 x 103 N/m
In quantitative analysis was done by measuring infrared absorbance of a mixture
containing compounds elaidat and oleat with a particular composition. Based on the
spectrum graph was obtained from the measurement of a mixture of oleic-elaidic acid the
composition of oleic-elaidic acid respectively 75%. This contain C=O groups with
different wave numbers. On the spectrum of elaidic acid is greater than oleic acid. This is
indicated by the peak of elaidic acid steeper than oleic. The composition of oleic and
elaedic in the mixturecan be calculated as follows:

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Known :

 C O olaedic  1712,79 cm -1
 C O oleiic  1743,65 cm -1

T0 elaedic = 68.3335 T0 oleic = 68.3335

Ts elaedic = 46.6668 Ts oleic = 39.16685
Answer:
a. Absorbance of elaidic:
A = -log T

T0
A  log
Ts
68.3335
 log
46.6668
 1.46
Absorbance of oleic:
A = -log T

T0
A  log
Ts
68.3335
 log
39.16685
 1.745

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Concentration of elaidic:

k
v  5.3x10 12
μ
k
1712.79  5.3x10 12
1.15x10 -26
k 1712,79

1.15x10 - 26
5.3x10 12
2
 k   1712.79 
2
    12 
 1.15x10 -26   5.3x10 
 
k
- 26
 1.04 x10 29
1.15x10
k  1.15x10 -26 x 1.04 x10 29  1196 N/m
A=k.C
1.46 = 1.196 x 103 N/m . C
C = 1.22 x 10-3 M
Concentration of oleic:

k
v  5.3x10 12
μ
k
1743.65  5.3x10 12
1.15x10 - 26
k 1743.65

1.15x10 - 26
5.3x10 12
2
 k   1743,65 
2
    12 
 1.15x10 - 26   5.3x10 
 
k
- 26
 1.08 x10 29
1.15x10
k  1.15x10 - 26 x 1.08 x10 29  1245 N/m
A=k.C
1.745 = 1.245 x 103 N/m . C
C = 1.4 x 10-3 M
Concentration of mixture= 1.22 x 10-3 M +1.4 x 10-3 M= 2.62 x 10-3 M

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b. The composition of Elaidat in Mixture
[elaidic]
% elaidic  x 100%
[mixture]
1.22 x 10 -3
 x 100%
2.62 x 10 -3
 46.56%
Composition of Oleat in Mixture

[oleic]
% oleic  x 100%
[mixture]
1.4 x 10 -3
 x 100%
2.62 x 10 -3
 53.44%
Based on the above calculations, the comparison of the composition elaedic and oleic acids in
the mixture are 46.56% and 53.44% respectively

 Quantitative Analysis in VCO

In the experiment instrument, infrared spectra, absorption measurements are also
performed on the VCO (Virgin Coconut Oil). Spectral measurements performed with a range of
500-4000 cm-1 vibration. The image measurement results are as follows.

Figure 3: infrared spectra VCO

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 Based on the spectrum, so:

FUNCTIONAL GROUP
WAVE NUMBER (cm-1)
BOND COMPOUND

725,23 C–H Alkene, aromatic rings

1111 C–O Alcohol, ether, carboxyclic acid, ester

1165 C–O Alcohol, ether, carboxyclic acid, ester

1365,6 NO2 Nitro Compound

1435,04 C–H Alkane

1743,65 C=O Aldehide, ketone, carboxyclic acid,

ester

2854,65 C–H Alkane

2924,09 C–H Alkane

Tabel 3. Wave number for VCO sample

Based on the data obtained can be calculated constants C=O bond in the following way.

Known: wave number  v  = 1743.65cm-1 and c = 3 x 108 m/s



 

asked : k =…….?
Answer:
Mass of carbon:
12 10 -3 kg / mol
m1  1 atom 2.0  10  26 kg
6.02  10 atom / mol
23

Mass of oxygen:
16 10 -3 kg / mol
m2   1 atom 2.7  10  26 kg
6.02  10 atom / mol
23

  1.15 10 26 kg

k
  5.3 10 -12

k
1743.65 cm 1  5.3 10 -12 s / cm
1.15 10  26 kg

10
 1743.65 cm 1 k

5.3 10 s / cm
-12
1.15 10  26 kg
k
329  1012 s 1 
1.15 10  26 kg
k  ( 329  1012 s 1 ) 2 (1.15 10 26 kg)
k  (108241 10 24 s  2 )(1.15 10  26 kg)
k  124477.15  10  2 kg s  2
k  1244.7715 kg s  2
k  1.245  10 3 N / m
So, constant bond of C=O in the liquid sample B is 1.245 x 103 N/m
From the value bond constant of C = O the concentration of VCO can be determined:
Known : k = 1.245 x 103 N/m
Ask : C =…….?
Answer:

Ts
A =- log
T0

17.5
A = - log
70
A = 0.602
A=kxC
C = A/k
C = 0.602 /1.245 x 103 N/m
C = 4.836 x 10-4 M
So, concentration of VCO is 4.836 x 10-4 M.

CONCLUSION
Based on the discussion above it can be concluded that the characteristic and organic compound
can be determined by knowing the frequency range. The solid sample concluded as ester whereas
liquid sample concluded as aldehyde.

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