1

1.0 ABSTRACT
Fourier transform infrared spectroscopy (FT-IR) is the favoured method for infrared detection of
chromatographically separated species. It is a device that splits a beam of radiation into two and
then recombines the two resultant beams after introducing a phase difference between them.
Ultraviolet and visible light (UV-vis) spectroscopy is essential optical methods for polymer
characterization. UV-vis is restricted to polymers that have specific chromophores such as aromatic
groups, conjugated double- or triple bonds, carbonyls or azo-subunits: When molecules absorb light
in the UV-vis range, electronic transitions are induced in the molecules between electronics levels
(commonly

and

transitions) in the most important range of the wavelength, that is

between 200 nm to 600 nm. The purpose of this experiment is to identify 12 compounds (A-L) using
your knowledge of UV-Vis and IR spectroscopy. Infrared (IR) spectroscopy is a technique used in
organic chemistry to identify the presence of certain functional groups in a molecule. The absorption
of UV or visible radiation relates to the excitation of outer electrons. There are 3 types of electronic
transition to be considered; 1) Transitions involving , and n electrons 2) Transitions concerning
charge-transfer electrons 3) Transitions relating to d and f electrons. To conduct both experiment,
samples of unknown compounds (of A-L) is scanned by using both (UV-vis) spectroscopy and FTIR
spectroscopy. Based on range of values of
max
, compound A is most likely be ethanol, compound B
is ethanol, compound C is ethanoic acid, compound D ethyl ethanoate, compound F is urea or
carbamide, G is benzene, H is toluene, I is benzoic acid, J is alkyl halide , K is diethyl ether and L is
cyclohexane. To avoid photokeratitis; for exposure to visible radiation, to avoid photoretinitis; and
for visible and IR radiation to avoid cataract after prolonged exposure and chorio-retinal injury from
low-luminance IR irradiation sources, the threshold limit values (or levels of exposure) for exposure
to UV radiation published by the ACGIH should be obeyed.















2
2.0 INTRODUCTION
2.1 Fourier Transform InfraRed
FTIR stands for Fourier Transform InfraRed Spectrometer or simply known as infrared spectroscopy.
It is applicable in such as organic synthesis, polymer science, petrochemical engineering,
pharmaceutical industry and food analysis.
FTIR is generally based on The Michelson Interferometer Experimental Setup; an example is shown
in Fig. 1. The interferometer consists of a Beam splitter, a fixed mirror, and a mirror that translates
back and forth, very precisely. The beam splitter is made of a special material that transmits half of
the radiation striking it and reflects the other half. Radiation from the source strikes the beam
splitter and separates into two beams. One beam is transmitted through the beam splitter to the
fixed mirror and the second is reflected off the beam splitter to the moving mirror. The fixed and
moving mirrors reflect the radiation back to the beam splitter. Half of this reflected radiation is
transmitted and half is reflected at the beam splitter, resulting in one beam passing to the detector
and the second back to the source. The radiation range for FTIR is at 4000 - 600

.



Figure 1 A schematic of a generic Michelson Interferometer

2.2 UV-Visible
Ultraviolet-visible spectroscopy or ultraviolet-visible spectrophotometer (UV-Visor UV/Vis) refers
to absorption spectroscopy in the ultraviolet-visible spectral region. This means it uses light in the
visible and adjacent (near-UV and near-infrared (NIR)) ranges. The absorption in the visible range
directly affects the perceived colour of the chemicals involved. In this region of the electromagnetic
spectrum, molecules undergo electronic transitions. This technique is complementary
to fluorescence spectroscopy, in that fluorescence deals with transitions from the excited state to
the ground state, while absorption measures transitions from the ground state to the excited state.
The radiation range is at 200-780nm.


3
UV-Vis spectroscopy is routinely used in analytical chemistry for the quantitative determination of
different analytes, such as transition metal ions, highly conjugated organic compounds, and
biological macromolecules. Spectroscopic analysis is commonly carried out in solutions but solids
and gases may also be studied.
Solutions of transition metal ions can be colored (i.e., absorb visible light) because d electrons within
the metal atoms can be excited from one electronic state to another. The colour of metal ion
solutions is strongly affected by the presence of other species, such as certain anions or ligands. For
instance, the colour of a dilute solution of copper sulfate is a very light blue;
adding ammonia intensifies the colour and changes the wavelength of maximum absorption
Organic compounds, especially those with a high degree of conjugation, also absorb light in the UV
or visible regions of the electromagnetic spectrum. The solvents for these determinations are often
water for water-soluble compounds, or ethanol for organic-soluble compounds. (Organic solvents
may have significant UV absorption; not all solvents are suitable for use in UV spectroscopy. Ethanol
absorbs very weakly at most wavelengths.) Solvent polarity and pH can affect the absorption
spectrum of an organic compound. Tyrosine, for example, increases in absorption maxima and
molar extinction coefficient when pH increases from 6 to 13 or when solvent polarity decreases.
While charge transfer complexes also give rise to colours, the colours are often too intense to be
used for quantitative measurement.

3.0 OBJECTIVE
The purpose of this experiment is to identify 12 compounds (A-L) using your knowledge of UV-Vis
and IR spectroscopy.

4.0 THEORY
4.1 FTIR
Infrared (IR) Spectroscopy is a technique used in organic chemistry to identify the presence of
certain functional groups in a molecule. IR lights are electromagnetic radiation with a wavelength
longer than visible lights. The molecule will cause a change in dipole moment when it undergoes
rotational or vibration when it is being absorbed by the IR spectroscopy.
Absorption occurs when the energy of the beam of light (photons) are transferred to the molecule.
The molecule will have a high energy forces that will cause it to get excited thus moves to a higher
energy state. The energy transfer will take place in the form of electron ring shifts, molecular bond
vibrations, rotations and translations. IR is commonly focusing on stretching and bending. The
movements of bending and stretching are as shown in Figure 2.


4

Figure 2 Movements of bending and stretching of molecules.

4.2 UV-Vis
The Beer-Lambert law states that the absorbance of a solution is directly proportional to the
concentration of the absorbing species in the solution and the path length. Thus, for a fixed path
length, UV/Vis spectroscopy can be used to determine the concentration of the absorber in a
solution. It is necessary to know how quickly the absorbance changes with concentration. This can
be taken from references (tables of molar extinction coefficients), or more accurately, determined
from a calibration curve.
A UV/Vis spectrophotometer may be used as a detector for HPLC. The presence of an analyte gives a
response assumed to be proportional to the concentration. For accurate results, the instrument's
response to the analyte in the unknown should be compared with the response to a standard; this is
very similar to the use of calibration curves. The response (e.g., peak height) for a particular
concentration is known as the response factor.
The wavelengths of absorption peaks can be correlated with the types of bonds in a given molecule
and are valuable in determining the functional groups within a molecule. The Woodward-Fieser
rules, for instance, are a set of empirical observations used to predict
max
, the wavelength of the


5
most intense UV/Vis absorption, for conjugated organic compounds such as dienes and ketones. The
spectrum alone is not, however, a specific test for any given sample. The nature of the solvent, the
pH of the solution, temperature, high electrolyte concentrations, and the presence of interfering
substances can influence the absorption spectrum. Experimental variations such as the slit width
(effective bandwidth) of the spectrophotometer will also alter the spectrum. To apply UV/Vis
spectroscopy to analysis, these variables must be controlled or accounted for in order to identify the
substances present.

The absorbance of a solution increases as reduction of the beam increases. Absorbance is directly
proportional to the path length, b, and the concentration, c, of the absorbing species. Beer's
Law states that
A = bc, where is a constant of proportionality, called the absorbtivity.
Different molecules absorb radiation of different wavelengths. An absorption spectrum will show a
number of absorption bands corresponding to structural groups within the molecule.

4.3 Electronic transitions
The absorption of UV or visible radiation corresponds to the excitation of outer electrons. There are
three types of electronic transition which can be considered;
1. Transitions involving , , and n electrons
2. Transitions involving charge-transfer electrons
3. Transitions involving d and f electrons (not covered in this Unit)
When an atom or molecule absorbs energy, electrons are promoted from their ground state to an
excited state. In a molecule, the atoms can rotate and vibrate with respect to each other. These
vibrations and rotations also have discrete energy levels, which can be considered as being packed
on top of each electronic level.

Absorbing species containing , , and n electrons


6
Absorption of ultraviolet and visible radiation in organic molecules is restricted to certain functional
groups (chromophores) that contain valence electrons of low excitation energy. The spectrum of a
molecule containing these chromophores is complex. This is because the superposition of rotational
and vibrational transitions on the electronic transitions gives a combination of overlapping lines. This
appears as a continuous absorption band.
Possible electronic transitions of , , and n electrons are;


4.4
*
Transitions
An electron in a bonding orbital is excited to the corresponding antibonding orbital. The energy
required is large. For example, methane (which has only C-H bonds, and can only
undergo
*
transitions) shows an absorbance maximum at 125 nm. Absorption maxima due
to
*
transitions are not seen in typical UV-Vis. spectra (200 - 700 nm)

4.5 n
*
Transitions
Saturated compounds containing atoms with lone pairs (non-bonding electrons) are capable
of n
*
transitions. These transitions usually need less energy than
*
transitions. They can
be initiated by light whose wavelength is in the range 150 - 250 nm. The number of organic
functional groups with n
*
peaks in the UV region is small.

4.6 n
*
and
*
Transitions
Most absorption spectroscopy of organic compounds is based on transitions of n or electrons to
the
*
excited state. This is because the absorption peaks for these transitions fall in an
experimentally convenient region of the spectrum (200 - 700 nm). These transitions need an
unsaturated group in the molecule to provide the electrons.


7
Molar absorbtivities from n
*
transitions are relatively low, and range from 10 to100 L mol
-1
cm
-
1
.
*
transitions normally give molar absorbtivities between 1000 and 10,000 L mol
-1
cm
-1
.
The solvent in which the absorbing species is dissolved also has an effect on the spectrum of the
species. Peaks resulting from n
*
transitions are shifted to shorter wavelengths (blue shift) with
increasing solvent polarity. This arises from increased solvation of the lone pair, which lowers the
energy of the n orbital. Often but not always, the reverse (i.e. red shift) is seen
for
*
transitions. This is caused by attractive polarisation forces between the solvent and the
absorber, which lower the energy levels of both the excited and unexcited states. This effect is
greater for the excited state, and so the energy difference between the excited and unexcited states
is slightly reduced - resulting in a small red shift. This effect also influences n
*
transitions but is
overshadowed by the blue shift resulting from solvation of lone pairs.

5.0 APPARATUS
1. FTIR spectrometer
2. Ultravioletvisible spectroscopy.


















8
6.0 PROCEDURE
6.1 Procedure for FTIR
1. Computer was turned on. Clicked at software-Spectrum.
2. Background was clicked and last Ready for scan sample was appeared.
3. Instrument-scan was clicked. The box was appeared. The information was keyed in.
4. Scan was clicked and the range to run sample was insert. Start to end.
5. Apply was clicked and scanned when sample was ready.
a. Top plate was cleaned with acetone.
b. Liquid sample sample was dropped into hole of top plate.
c. Solid sample Sample was inserting into hole and was purged. Do not purge more
than 60.
d. Sample was ready to scan.
e. Top plate was cleaned again with acetone after used.
6. Edit was clicked the result was copied into Microsoft Excel.
7. Exit was clicked to log out from program.

6.2 Procedure for UV-Vis
To turn on equipment
1. Instrument was turned on. It took approximately 5 minutes to warm up.
2. Software Perkin Elmer UV VIS/NIR was turned on.
3. Went to New and Method was clicked.
4. Type of instrument was selected (high performance UV VIS/NIR) and then the lambda was
chose for your instrument (Lambda 750).
5. Next was clicked and method type was selected.
6. After that, scanned and next was clicked (twice).
7. Save/Finish and named of method then ok was clicked.
To create method
1. Went to data collection.
2. Wavelength was set depending on manual lab/test (ex: from 300nm until 500nm)
3. Went to ordinate mode and the parameter was chose (absorption, transmittance or
reflection).
4. Went to sample info then the quantity of sample was insert, hit Enter.
5. The first sample was put as a blank.
6. The sample Id was edited with other samples (ex: bromide 1, bromide 2 and etc).
7. The wavelength that was desired to start was set like before this, then apply was clicked and
ok.
8. Button start was clicked.
9. Ok was clicked.
10. Both with blank in the sample holder were insert, the clear windows aligned with the optical
path.


9
11. Blank at beam no. 2 was removed, replaced with the other sample and ok was clicked.
12. Then, we got the spectrum.
13. The procedure no. 4 was repeated until all sample finished.
To save data
1. Went to file, save as was clicked.
2. Task was clicked and name of the file was insert and was saved.
3. After that went to Processing, Add was clicked then what process was selected (ex: peak
table).
4. Went to report, template was clicked. Type of report that was chose (ex: scan with peak
table) then saved.
5. Went to the icon printer report and thick printer file.
6. We chose where we want to save our file then saved.
To shutdown method
1. Went to report and file was clicked.
2. Exit and yes was clicked.
3. Equipment was off, PC was shut down and lastly power supply.


















10
7.0 RESULT
Table 1: The identification results from experimental of FTIR and UV-Vis.
Unknown
compound
(sample)
UV-Vis

max
(nm)
IR band
(cm
-1
)
Descriptions of
functional groups and
its bonding
Name of the compound
and its structure
A 210.02 3041.12
2630.40
Alcohol(O-H)
Alkane(C-H)
Ethanol


C 210.19 2634.90
2630.64
1757.25

Alkane(C-H)
Acid(O-H)
Carbonyl(C=O)
Acetic acid/Ethanoic
acid
(Carboxylic acid)



D 210.09 2984.67
1233.60
1096.8
Alcohol(O-H)
Ester(C=O)
Ether(C-O)
Ethyl ethanoate
(Ester group)

J 210.32 559.55

Alkyl Halide(C-Br) Bromoethane
(Alkyl Halide group)



K 210.03 2976.59
1297.13

Alkane(C-H)
Ether(C-O)
Diethyl ether
(aldehyde group)

I 227.62 3071.10
2669.54
1916.17
Aromatic(C-H)
Acid(O-H)
Alkene(C=C)
Benzoic acid







11
Table 2: The identification results from the given data in lab manual.
Unknown
compound
(sample)
UV-Vis

max
(nm)
IR band
(cm
-1
)
Description Name of functional
group and its structure
B
max
= 289 nm
log10 = 1.19

1730 cm
-1
Aldehydes (C=O) Ethanal (Acetaldehyde)

E
max
= <210,
220 and 273
nm
log10 = >4,
3.86 and 2.86
2950 cm
-1

1685 cm
-1


Alkanes (C-H)
Alkenes (C=C)
Carboxylic acids (C=O)

Acetylsalicylic acid
(Aspirin)



F
max
= <220 nm
log10 = >3.2
3430 cm
-1

1592 cm
-1

Amines (N-H)
Nitro compounds
(NO
2
)
Urea (Carbamide)



G
max
= <178 nm
log10 = 4.84
3037 cm
-1

1480 cm
-1


Aromatic ring (C-H)
Aromatic rings (C=C)

Benzene



H
max
= 260 nm
log10 = 2.43

3033 cm
-1

1501 cm
-1

730 cm
-1


Aromatic ring (C-H)
Aromatic rings (C=C)
Alkenes (C-H)

Toluene


J

-
958.74 cm
-1


Alkenes (C-H)

Bromoethane



L

-
2900 cm
-1

1463 cm
-1


Alkanes (C-H)
Alkanes

Cyclohexane





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8.0 DISCUSSION
8.1 Compound A
It has been determined that sample A is from the alcohol group. In the FTIR, the peak of 3041.12 cm
-
1
from the spectrum has been recorded and showed that the compound contains OH bonding. It
can easily be detected because for alcohol group no matter of what alcohol molecule involves this
very broad peak of O-H will appear. Another peak obtained is 2630.4 cm
-1
, which states the
functional group is alkane, (C-H). Both alcohol and alkane group is stretching. From the result in UV-
VIS, it showed that compound A contains chromophore which act as absorbing group. Therefore the
mostly compound that have those value is an ethanol.

8.2 Compound B
The functional group for compound B can determined by the value of
max
and and IR which is 289
and 1730 cm
-1
respectively. From the value, it showed that compound B is ethanal or in aldehyde
group. The value of
max
and absorptivity indicates that the compound contain chromophore. The IR
value indicates that C=O is stetching. Therefore, from the information given it showed that the
compound is ethanal.

8.3 Compound C
Compound C is determined as ethanoic acid. In the FTIR spectrum, the peak value 1757.25 cm
-1

showed the carbonyl group, C=O and the compound is stretching. The intensity of a compound is
strong. The other peak in the spectrum is 2630.64 cm
-1
which showed the alkene group, C=H in the
compound. The IR value of 2634.90 cm
-1
showed O-H bonding which indicates that the functional
group of this compound is carboxylic acid. UV-Vis result also indicates that the compound contains a
chromophore.

8.4 Compound D
Compound D is identified to be ethyl ethanoate. According to IR results, it shows that the compound
has peak of 1233.60 cm
-1
which indicates that C=O of ester is stretching. The other peak with value
of 2984.67 cm
-1
means that presence C-O of alkane is stretching. From UV-Vis analysis, the
compound observed indicates present of chromophore with a
max
of 210.09nm. Moreover,
compound D identification also can be determined based on the equation given in the lab manual
which is:


As had been discussed earlier, compounds A and C are identified to be pentanol and
carboxylic acid respectively. An alcohol reacts with carboxylic acid to give esters a reaction that is
common in both laboratory and living organisms. Esters are produced when carboxylic acids are
heated with alcohols in the presence of a concentrated sulphuric acid catalyst This reaction is called
esterification reaction.



HCl


13
8.5 Compound E
Compound E can be determined easily from the value of UV-Vis and IR that existing in the lab
manual sheet. The compound is found out to be acetylsalicyclic acid. The range values of
max
in UV-
Vis is <210,220 and 273 and log
10
= >4, 3.86 and 2.86. in addition, the IR value is 2950 and 1685 cm
-
1
. From the values, the functional group of alkanes, (C-H), alkenes (C-C) and carboxylic acids (C=O)
are determined. IR spectrum value of 2950 cm
-1
for peak one is C-H alkane stretch. Other peak value
is 1685 cm
-1
that stand for C=O carboxylic acid stretching. The result in UV-Vis showed that the
compound contains chromophore.

8.6 Compound F
Compound F is easy to determine because the value of IR and UV-Vis are given in the lab manual.
The
max
and log
10
of compound F are <220 and >3.2 respectively. UV-Vis detects that compound
having chromophores. IR result showed that the peak values are 3430 cm
-1
and 1592 cm
-1
. The peak
of 3430 cm
-1
is a group of amines or amides, N-H and 1592 cm
-1
aromatic ring, C=C stretching. From
the result, it shows that the compound is urea or carbamide.

8.7 Compounds G, H and I
Based on the information given, compound G is observed to be benzene. The IR band shows that the
first peak is at 3037cm
-1
which clearly indicates that C-H of aromatic ring is stretching. Another peak
is at 1480cm
-1
shows that there is C=C aromatic ring is stretching. On the other hand, compound H is
identified to be a toluene. From IR results, there are three peaks given which are 3033, 1501, and
730cm
-1
. These indicate that there are aromatic rings stretching in first and second peak and C-H of
alkenes stretching at the third peak. Both of G and H compounds contains chromophore as both UV-
Vis results shows that they have
max
of 178 and 260nm respectively.
For compound I, it is identified as a benzoic acid. From IR result shows that there is four
peaks exist in the spectrums which are at 3071.10, 2669.54, and 1916.17 cm
-1
. All four peaks indicate
that there are aromatic, acid, acyl halide and alkene functional group presented in the compound.
Based on the UV-Vis results, the compound observed contains chromophores. To be proved, the hint
given in lab manual is used to ensure that the all identification is true. Based on the equation:





The first reaction which is from compound G converts to H is called Friedel-craft alkylation
where an alkyl group can be added to a benzene molecule by an electrophile aromatic substitution
reaction to form alkyl benzene [3]. For instance, a benzene react with bromochlorine would form a
toluene:

14
The second reaction involves the oxidation process of compound H which an oxidizing agent
potassium permanganate,

. Compound H which is toluene will react with

to from a
benzoic acid.





8.8 Compounds J and K
Based on the data given for compound J, the IR band states that the peak formed is at the fingerprint
which is 958.74. In general, CX of alkyl halide vibration frequencies appear in the region 850-515
cm
-1
, sometimes out of the range of typical IR instrumentation. According to the experimental
results of FTIR, the peak shows at 559.15 cm
-1
indicates that there is a stretching present in C-Br of
alkyl halide. For compound K, there are two peak observe in IR spectrum which are 2976.59 and
1297.13 cm
-1
which indicates the present of C-H of alkane and C-O of ether. Therefore, the
compound K is observed to be diethyl ether. Since both compounds are single bond, the UV-Vis
spectrum is negligible. The hint given in the lab manual is referred as a confirmation of the
compounds identity.



This reaction is involve between an alkoxide with a primary haloalkane reacts which a
present of a strong base to form ether [4]. The reaction called Williamson Ether Reactions.


8.9 Compound L
Compound L can be determined by the IR value given in the lab manual. The peak given is 2900 and
1463 cm
-1
. Both of peak showed that the functional group is alkane, C-H. At 2900 cm
-1
the type of
vibration is stretching while at 1463 cm
-1
the type of vibration is bending. From the result, it showed
that the compound is cyclohexane.








15
9.0 CONCLUSION
In brief, the unknown compounds are identifiable by using ultraviolet and visible radiation and FTIR.
Compound A is most likely be ethanol, compound B is ethanol, compound C is ethanoic acid,
compound D ethyl ethanoate, compound F is urea or carbamide, G is benzene, H is toluene, I is
benzoic acid, J is alkyl halide , K is diethyl ether and L is cyclohexane.

10.0 RECOMMENDATIONS
The threshold limit values (or levels of exposure) for exposure to UV radiation, to avoid
photokeratitis; for exposure to visible radiation, to avoid photoretinitis; and for visible and IR
radiation to avoid cataract after prolonged exposure and chorio-retinal injury from low-luminance IR
irradiation sources published by the ACGIH should be obeyed.

11.0 REFERENCES
1. Theory of Ultraviolet (UV-Vis) Spectroscopy. (nd). Retrieved from
http://www.chem.ucla.edu/~bacher/UV-vis/uv_vis_tetracyclone.html.html
2. Visible and Ultraviolet Spectroscopy. (nd). Retrieved from
http://www2.chemistry.msu.edu/faculty/reusch/virttxtjml/spectrpy/uv-vis/spectrum.htm
3. FTIR Spectroscopy Basic Theory. (nd). Materials and Engineering Research Institute Sheffiled
Hallam University. Retrieved from https://www.shu.ac.uk/research/meri/research/ftir-
spectroscopy-basic-theory