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ABSTRACT

In this experiment we are going to identify organic compounds by using UV-Vis and IR Spectroscopy. The
purpose of this experiment is to provide an opportunity to implement the skills have gained from the first
two chapters of the Instrumental Chemistry course. We will identify 12 compounds (A-L) using knowledge
of UV-Vis and IR Spectroscopy. The experiment was conducted by testing 6 unknown compounds by using
UV-Vis and IR Spectroscopy. Each compounds has different value of frequency range, by comparing the
data we get from experiment with information given by lecturer all the compounds can be determined. The
unknown compounds are A = Ethanol, C = Ethanoic Acid, D = Ethyl Ethanoate, I = Benzoic Acid, J = Ethyl
Bromide and K = Diethyl Ether. The experiment is successfully done with some errors that may influence
the result.

INTRODUCTION
Ultraviolet Spectroscopy (UV-Vis)
Ultraviolet (UV) was discovered more than 200 years ago by Johann Ritter. In 1801, he reported in Annalen
de Physik that, on the 22nd February, he had detected solar radiation. This is referring to Oldenburg and
Eden1 in the Ultraviolet Spectroscopy and UV Lasers.
According to John and Stephen2, the infrared portion of the electromagnetic spectrums spans from about
3.8x1014 Hz to 6 x1012 Hz. If the sample is illuminate by radiation of a particular and discrete wavelength,
then absorption of that radiation might occur. Most of the common functional group absorbs IR radiation
within the range 200 780 nm. At other wavelength the radiation might not occur.

Figure 1: Lambda 750 UV/Vis/NIR Spectrophotometer (Annon, 2013)3


In this experiment, the Lambda 750 UV/Vis/NIR Spectrophotometer is designed to give highest sampling
flexibility especially it is commonly use in Chemistry, Forensic and Applied Science field. This system is
suitable for research or teaching laboratory, as accessories and detector modules can quick and easily
interchanged to provide the optimum configuration for each measurement. This instrument also maintains
critical performance at the highest level.
The main purpose of this instrument is to analyze compounds in the ultraviolet and visible regions of
electromagnetic spectrum. The degree of conjugated double or triple bonds and degree of aromaticity is the

Oldenburg, A. L ,Eden, J. G.,(2002), Ultraviolet spectroscopy and UV laser,371-477


John C. Gilbert, Stephen F. Martin.,(2002), Experimental organic chemistry : A miniscale and
microscale approach
Edition,Brooks/Cole Thomson Learning.
2

Anon,(2013). http://www.perkinelmer.com. Retrieve on 3 April 2013.

structure of binders which are evaluated by using UV-Vis spectroscopy reported by Mills and George D 4.
The UV radiation is passed through the sample and absorption radiation occurs. The radiation transmitted
is detected by photocells and the spectrometer records the absorption by comparing the difference
between the intensity of the radiation passing through and the reference cells. Nowadays, the UV-Vis
spectroscopy is being upgraded to give better results and expanding the potentials of the UV-Vis field.

IR Spectroscopy
Fourier Transform Infrared or known as FTIR is a method of infrared spectroscopy that can be used to
identify chemicals that are either organic or inorganic and also can be used on solids, liquids, and gases. It
also can measure all the Infrared (IR) wavelengths simultaneously which this feature is called the Multiplex
or Felgett Advantage and produces a full spectrum. Simply, it can absorb different measurement of the IR
frequencies by positioned a sample in the path of an IR beam.

Figure 2: Perkin Elmer FTIR Spectrum 100


In this experiment, the Perkin Elmer FTIR Spectrum 100 is used this instrument provides the highest quality
lab experience, along with the most accurate and reproducible results in the industry.
The main objective of the FTIR spectroscopic analysis is to identify of compounds by matching spectrum of
unknown compound with reference spectrum (finger printing). Moreover, it also can be used to detect
functional groups in unknown substances.
In addition, this spectroscopy can produce an infrared absorption spectrum that is like a molecular
"fingerprint" to identify the types of chemical bonds in a molecule. Just like the fingerprint, no two unique
molecular structures can produce the same infrared spectrum so this will make the infrared spectroscopy
useful for several types of analysis.

AIMS
In this experiment, we are going to
1. Identify organic compounds with are A, C, D, I, J and K by using UV-Vis and IR Spectroscopy.

Mills and George, D.,(2012), Ultraviolet/visible spectroscopy. http://www.astm.org. Retrieve


on 16 March 2013.

THEORY
The ultraviolet-visible (UV-Vis) spectrophotometer is an instrument commonly used in the laboratory to
analyze compounds in the ultraviolet (UV) and (Vis) regions of the electromagnetic spectrum. The
wavelength of UV is from 200 380 nm while wavelengths for the UV-Vis are fall between 200 and 780 nm.
It is known that, the shorter the wavelength, the higher the energy.
The UV-Vis spectroscopy works by looking at the electrons transition. The electrons transitions of a
compound can determine the wavelength and the maximum absorbance of compounds. The UV or visible
radiation is absorbed due to the excitation of the outer electrons. The electrons are promoted from their
ground state to an excited state when an atom or molecule absorbs the energy. The atoms can rotate and
vibrate with respect to each other in a molecule. The vibrations and rotations also have discrete energy
levels, which can be said as being packed on top of each electronic level. The absorption of the ultraviolet
and visible radiation in organic molecules is restricted to several functional groups (chromophores) which is
contains valence electrons of low excitation energy. The spectrum of a molecule containing the
chromophores is complex that is because the superposition of rotational and vibrational transitions gives a
combination of overlapping lines on the electronic transitions. This will show as a continuous absorption
band.

Figure 1: Possible electronic transitions of p, s, and n electrons


* Transitions
This transition is where the anti-bonding orbital is being excited by an electron in a bonding s orbital. The
energy required is large. For example, methane (which has only C-H bond, and can only undergo
* transitions) that shows an absorbance maximum at 125 nm.
n * Transitions
This transition is capable for the saturated compounds containing atoms with lone pairs (non-bonding
electrons). These transitions usually need less energy than * transitions. The light whose have the
wavelength that is in the range from 150 to 250 nm can only initiated this transition.
n * and * Transitions
The transitions of n or p electrons to the p* excited state is the most absorption spectroscopy of organic
compounds that being happened. It was been said like that because the absorption peaks for these
transitions fall in an experimentally convenient region of the spectrum (200 - 700 nm). Other than that,
these transitions also need an unsaturated group in the molecule to provide the p electrons.
The core principle used behind the absorbance in this spectroscopy is Beer-Lambert Law (Equation1). For
a single wavelength, the symbol of A is the absorbance (it is unitless and usually seen as arbitrary units),

is for the molar absorptivity of the compound or molecule in solution (M-1cm-1), b is for the path length of the
cuvette or sample holder (1 cm long), and c is the concentration of the solution (M).
A = bc

Equation (1)

However, the UV-Vis spectroscopy can identify the chromophores or known as functional group of the
compound only. While fourier transform infrared spectroscopy (FT-IR) can identify the specific functional
group for the sample of compound. The region where the specific chromofores can be detected is between
400 and 600 cm-1 .This FT-IR is a method of infrared spectroscopy that can be used to identify chemicals that
are either organic or inorganic and also can be used on solids, liquids, and gases.
Modern instruments are FTIR spectrometer that obtains the IR spectrum by Fourier transformation of the
signal from an interferometer with a moving mirror. (Sawyer et al., 2008). According to Gable (2013) the
mathematical expression of Fourier transform can be expressed as
F() =
And the reverse Fourier transform is
f(x) =
Where is an angular frequency and x is the optical path difference in our case. F() is the spectrum and
f(x) is called interferogram.

Figure 1.1 : Michelson


2006)

interferometer

experiment

In
a
consists of

interferometer

adapted

Michelson

for

(Anon,
FTIR

Stationary mirror
Moving mirror
Beamspitter
Detector
Sample

The working principle of Michelson Interferometer


1. Source of energy is send through an interferometer and onto the sample. All source radiation gets to
the sample.
2. The light passes through a beamsplitter, which sends the light in two directions at right angles. One
beam goes to a stationary mirror then back to the beamsplitter. The other goes to moving mirror.
3. The motion of the mirror makes the total path length variable versus that taken by the stationarymirror beam. When the two meet up again at the beamsplitter, they recombine, but the difference
path in path lengths creates constructive and destructive interference.
4. The recombined beam passes through the sample. The sample absorbs all the different
wavelengths characteristic of its spectrum, and this subtracts specific wavelengths from the
interferogram
5. The detector now reports variation in energy versus time for all wavelengths simultaneously.

6. A laser beam is superimposed to provide a reference for the instrument operation.

APPARATUS & CHEMICALS

UV-Vis Spetrometer

FT-IR Spectrometer

6 unknown compounds labelled


as A, C, D, I, J, and K.

Dropper

PROCEDURE
UV-Vis
To Turn on the Equipment
1.
2.
3.
4.

The instrument is turned on. It took 5 minutes to warm up.


The software Perkin Elmer UV VIS/NIR is turned on.
Went to New and Method is clicked.
Type of instrument (high performance UV/VIS/NIR) is selected and clicked Next and the wavelength
for the instruments is chosen.
5. Clicked Next and the method type is selected.
6. After that, scanned and clicked Next twice.
7. The method is saved and named. Then, clicked ok.
To Create Method
1.
2.
3.
4.
5.

Went to the data collection.


The wavelength is set depending to manual lab which is from 300 nm to 500 nm.
Went to ordinate mode and the Absorbance parameter is chosen.
Went to sample info then the quantity of sample is inserted, ENTER is pressed.
For the first sample, it was put as blank.

6.
7.
8.
9.

The sample ID is edited with A, C, D, I, J and K.


The wavelength is set for the experiment.
Clicked button start and then ok.
The sample holder is inserted with both blank sample which is contains ethanol. The fingerprints
around the glass container where the sample is filled in.
10. Next, repeat steps 8 and 9 by replaced one of the ethanol sample with sample A, C, D, I, J and K.
11. For the sample of solid I, the sample is diluted first before placed in the sample holder.

FTIR spectroscopy
1. 5 solutions of samples and 1 solid sample is prepared.
2. The computer is turned on. Clicked at software spectrum.
3. Clicked at the background and Ready for scan sample is appeared.
4. Clicked at instrument scan. The box is appeared where the information need to be key in.
5. Clicked at scan and the range to run the sample is inserted.
6. Apply is clicked and the sample is scanned when it is ready.
7. The top plate is cleaned with acetone.
8. If the sample is liquid, it is drop into hole of top plate.
9. If the sample is solid sample, it is inserted into hole and it is purged.
10. The first sample to be scanned is sample A.
11. The sample is scanned.
12. After the first sample, the top plate is cleaned with acetone.
13. Step 4 to 12 is repeated with other samples.
14. Every results is copied into Microsoft Excel.

RESULTS

IR Spectra

Compound A

100.3

3712.15

95

3672.76

90
1273.96
1329.39

85

803.23

2926.28

2881.99

80

1379.60

75

622.25
3319.21

70

2973.13

% T 65

60

879.97
1087.70

55

50

45

40

1045.93

35

30.5
4000.0

3600

Compound C

3200

2800

2400

2000

1800
cm-1

1600

1400

1200

1000

800

600 515.0

99.9

95

3711.10

90
2555.89

85

2630.14

80
1051.83

75

2933.05
3039.82
1360.70
1756.99

70

1012.62

65
928.55
888.75

60
%T
1407.85

55

604.55

50

1231.71

45

626.63

40
1287.76

35

30

25
1704.49

20
17.0
4000.0

3600

Compound D

3200

2800

2400

2000

1800
cm-1

1600

1400

1200

1000

800

600 515.0

100.2

95

2086.28
564.81
528.08

3712.07
3648.98

90

917.41
786.06
2984.71

1447.67

1300.54

85
938.15
1097.89

846.83

80
634.30
607.72

75

70

65

60
%T
55
1372.78

50

45

40

35

30
1043.58

25
1736.53

1233.50

20
17.0
4000.0

3600

Compound J

3200

2800

2400

2000

1800
cm-1

1600

1400

1200

1000

800

600 515.0

97.4
96
94
92

3711.68
3648.86
2866.70

90
2924.17

88

2978.81

1377.22

86

768.68

84

1442.47

82
80
78
% T 76
74
72
70
958.78

68
66
64

1241.01

62
60
58
559.05

55.6
4000.0

3600

Compound K

3200

2800

2400

2000

1800
cm-1

1600

1400

1200

1000

800

600 515.0

97.4
95
3711.67
1491.04

90

1297.25

794.84
934.79

2805.66
1444.69

85

845.39
2933.03
1043.80

80
1350.55

75

1381.91
2860.32

70

1076.17

2976.37

65
%T
60

55

50

45

40

35

30

1118.25

27.5
4000.0

3600

Compound I

3200

2800

2400

2000

1800
cm-1

1600

1400

1200

1000

800

600 515.0

98.6
96

3735.03

1983.75

94

1600.72
1582.28

92

1323.19

1026.21

803.11

1682.51

90

1453.05
930.07

88

1360.83
1289.52

86

682.85
546.01

1072.59

84

665.96
703.83

1393.07

82
80
78
76
74
% T 72
70
2988.99

68
66

2870.10

64
62
60
58
56
54
52
50
48
47.0
4000.0

1142.59

3600

3200

UV-Vis Spectra

2800

2400

2000

1800
cm-1

1600

1400

1200

1000

800

600 515.0

FTIR analysis
Unknown
compound
(Sample)

Band ( IR)

3319.21

O-H stretch

Compound : Ethanol

2973.13

C-H stretch

presence of alcohol group.

2926.28

C-H stretch

2881.99

C-H stretch

3039.82

O-H stretch

2933.05

O-H stretch

1756.99

C=O stretch

1704.49

C=O stretch

2984.71

C-H stretch

1736.53

C=O stretch

1233.50

C-O stretch

1043.58

C-O stretch

2988.99

O-H stretch

Compound : Benzoic Acid

1983.75

Aromatic C-H stretch

(C6H5COOH)

1682.51

C=O stretch

presence of aromatic ring and


carboxylic acid group.

(cm-1)

Descriptions

Name of compound
and functional group

Compound : Acetic/ Ethanoic


acid (CH3COOH)
presence of carboxylic acid
group.

Compound : Ethyl ethanoate


(CH3COOHCH2CH3)
presence of ester group.

2978.81

C-H stretch

Compound : Ethyl bromide

2924.17

C-H stretch

presence of alkyl halide group.

2866.70

C-H stretch

1241.01

C-H wag

559.05

C-Br stretch

2976.37

C-H stretch

2860.32

C-H stretch

1118.25

C-O-C stretch

Compound : Diethyl ether

presence of ether group.

Uv-vis analysis

Compounds

max (nm)

Intensity
(A)

Absorptions
occurrence

Descriptions

Blank

No

213.25

0.2338

No

Double bond absent

214.09

0.856

Yes

Double bond present

214.13

1.039

Yes

Double bond present

215.11

3.173

Yes

Double bond present

214.23

1.603

No

Double bond absent

213.37

0.622

No

Double bond absent

DISCUSSION
In UV-Vis analysis, the blank that we used was water which has an approximately 180
nm cut-off wavelength. Cut off wavelength is the approximate wavelengths below which it
cannot be used because the solvent itself absorbs all of the light (Skoog, et al., 2007). The
cuvette bottle has a cut-off wavelength too which for glass and plastic were ranged between

380-780 nm (Barron, 2010). On the other hand, acetone is used for cleaning the cuvettes and
the plate because it is more effective than water to remove the traces of any impurities and oily
marks on the surfaces including the fingerprints. Acetone has a higher volatility than water too.
We can find the chromophores for each sample from the UV-Vis analysis by looking at
the wavelength. For samples C, D and I, they are having double bond in the structures. While
the others, samples A, J and K does not have double bonds. In FTIR analysis, we can examine
the stretching and bonding of the compounds from the wavelength that were recorded too.
For sample A, the infrared spectrum of the compound shows that the major absorptions
are found at 3319.21 cm-1 and 2973.13 cm-1. These correspond to the OH and CH stretch.
From this, we have expected that compound A is ethanol, CH3CH 2OH. As compound A do not
has double bonds, thus it was not detected by the instrument. The peak obtained was 213.25
nm and compound A undergoes an n* transition at max = 180 nm. This was proven below.
The reaction given in the information is :

PCC
CH2Cl2

CrO3

H2SO4
Reaction 1

We know that compound B is ethanal, CH 3CHO from the infrared spectrum of 1730 cm -1
and undergoes an n* transition with max = 289 nm. The reagent, Pyridinium chlorochromate
(PCC) and dichloromethane, CH2Cl2 is a reaction of oxidation as below,

Equation 1 Oxidation of Alcohol

For sample C, the infrared spectrum of the compound shows that the major absorptions
are found at 3039.82 cm-1 and 1704.49cm-1. These correspond to the OH and C=O stretch.
From this, we have expected that compound C is ethanoic acid, CH3COOH. The peak obtained
was 214.09 nm and compound C undergoes an n* transition. From equation 1, we can see
that oxidation of aldehyde can form a carboxylic acids. That means that compound B, CH 3CHO
form compound C, CH3COOH.
For sample D, the infrared spectrum of the compound shows that the major absorptions
are found at 1736.53cm-1 and 1233.50 cm-1. These correspond to the C=O and C-O stretch.

From this, we have expected that compound D is ethyl ethanoate, CH3COOHCH2CH3. The peak
obtained was 214.13 nm and compound D undergoes an np* transition. This was proven
below.The second reaction given in the information is as follows,

A+ C

HCl

Reaction 2

From the above reaction, we know that ethanol reacts with ethanoic acid to produce
compound D. In the presence of concentrated hydrochloric acid, HCl as a catalyst, reaction
between alcohol and carboxylic acid will produce an ester (Clark, 2002). In this case, compound
D will be CH3COOHCH2CH3.
Next, we have been given the following reaction :

CH3Cl
FeCl3

KMnO4
H2O

Reaction 3

We know that compound G is benzene, C 6H5 form the infrared spectrum of 3037 and
1480 cm-1 and undergoes an pp* transitions with max = 178 nm. From the reaction 3 above, we
can decide what is compound represented by H. From the reaction of Friedel-Crafts alkylation
with chloromethane, CH3OH in the presence of iron (III) chloride, FeCl 3 as catalyst, benzene will
turn to toluene, C6H4CH3 which is compound H. Compound H form infrared spectrum of 3033,
1501, and 730 cm-1 and undergo pp* transitions with the max = 260 nm.
For sample I, the infrared spectrum of the compound shows that the major absorptions
are found at 2988.99, 1983.75 and 1682.51 cm-1. These correspond to the OH and aromatic CH and C=O stretch. From this, we have expected that compound I is benzoic acid, C6H5COOH.
The peak obtained was 215.11 nm and compound I undergoes * and n* transitions.
From reaction 3, we can see that compound I is produced from the oxidation reaction between
compound H and the potassium permanganate, KMnO4. We know that KMnO4 oxidizes the
methyl group in toluene to form an aromatic carboxylic acid. Therefore, our expectation that
compound I is C6H5COOH was true.

A + J

NaH
THF
Reaction 4

For the last reaction is the reaction where sodium hydride in the reaction is a strong
base that always be used in a nucleophilic substitution reaction between alcohols with an alkyl
halide to produce ether. The reaction above provides the hint to determine compound J and K.
We already know that A is ethanol. So, compound J should be the alkyl halide and compound K
should be the ether. Only one alkyl halide is available in the list of possible compounds. Thus,
compound J is bromoethane or also known as ethyl bromide. Compound J forms the infrared
spectrum of 559.05 and 2978.81 cm-1 which represent C-Br and C-H stretch respectively. The
peak obtained was 215.11 nm. Ethyl bromide does not have any double bond.
From reaction 4, we can recognize that, compound K is an ether. The infrared spectrum
of the compound shows that the major absorptions are found at 1118.25 cm-1. This corresponds
to the C-O-C stretch. Thus, compound K is diethyl ether, (C 2H5)2O. The peak obtained was
213.37 nm and compound K undergoes n* transition at max = 180 nm which beyond the UVvisible region.

CONCLUSION
This experiment was carried out in order to identify 6 unknown organic compounds via the FTIR
and UV-Vis analysis. Based from the experiment, compound A is an alcohol (ethanol),
compound C is a carboxylic acid (ethanoic acid), compound D is an ester (ethyl ethanoate),
compound I is an aromatic carboxylic acid (benzoic acid), compound J is an alkyl halide (ethyl
bromide), and compound K is an ether (diethyl ether). The data was analysed based on the
peaks appeared both on the IR and the UV-Vis spectra. The presence or absence of functional
groups in all of the compounds could be detected by using the IR spectroscopy. The functional
group can be known by analysed the peaks of the spectra. While the UV-Vis spectroscopy only
analysed the compounds that contain at least one double bond. The results are in good
agreement with the information found in the literature. However, there might be a few
differences from the experiment. There might be impurities in some of the samples which finally
affected the resulting peaks. This could be caused by handling the laboratory equipment and
procedures carelessly and other reasons referred to in the discussions section. In IR
spectroscopy, the obtained spectra are resulted from the absorption of light when the radiant
energy matches the energy of a specific molecular vibration. While the UV-Vis spectra
correspond to the absorption that occurs during the transition of an electron from the ground
state to one of the many vibrational and rotational energy states. Besides the minor error in the
results, this experiment has been a success since the aim of the experiment was achieved and
we gained knowledge regarding the IR and UV-Vis spectroscopy.

RECOMMENDATIONS
There are several recommendations that can be deduced from this experiment to
improve the results of this experiment. While handling the volatile compounds, it is

recommended to dilute it with the solvent in a sample bottle with stoppered to avoid the sample
from being vaporized.
The amount of compound being diluted can also be the argument. It is possibly
inaccurate using the dropper to transfer a sample portion into the sample bottle. Therefore, we
can use automatic or digital pipette that can increase the reliability in the accuracy of the sample
amount.
When conducting the UV-Vis analysis, the cuvettes should be handling with care. The
cuvette should be held on the rough sides in order to make sure there is no fingerprint on the
smooth sides that would probably affect the resulting peak. The outer of the cuvettes also must
be wiped gently with tissue so that the traces of spills can be eliminated and to avoid any
scratch on the smooth sides.
While conducting the IR analysis, the sample plate should be cleaned with acetone. The
plate should be wiped gently with tissue in order to prevent the instantaneous damage because
the plate is very sensitive. Dropping the samples onto the plate must be done immediately
because if dropping too slowly the volatile sample will vaporize and the result will not be
accurate.
In this experiment, we have 6 unknown organic compounds. Since all the compounds
are colourless, it is necessary to label the sample bottles to avoid the mismatch information of
the compound with the obtained results.

REFERENCES
1. Andrew
R
Barron,
2010,
http://cnx.org/content/m34525/latest/

Basic

of

UV-Visible

Spectroscopy

2. D. A. Skoog, F. J. Holler, S. R. Crouch, Principles of Instrumental Analysis, 6th Ed.,


Thomson Brooks/Cole (2007).
3. Clark,
J.,
2002.
The
Mechanism
for
the
http://www.chemguide.co.uk/physical/catalysis/esterify.html

Esterification

Reaction

4. Bruice, P. Y., 2002. Chapter 13 : Mass Spectrometry and Infrared Spectroscopy. In:
Organic Chemistry. 4 ed. USA: Prentice-Hall, Inc., pp. 482-525.
5. Doudlas A.Skoog, F.James Holler,Stanly R.Crouch,Principles of Instrumental Analysis
Sixth Editon,2007, Chapter 16(430-443)
6. FTIR Instrumentation. Adopted from the FTIR lab instruction by H.-N. Hsieh, New Jersey
Institute of Technology http://www-ec.njit.edu/~hsieh/ene669/FTIR.html
7. FTIR Instrumentation. Adopted from the FTIR lab instruction by H.-N. Hsieh, New Jersey
Institute of Technology
http://www.chemguide.co.uk/physical/catalysis/esterify.html