TABLE OF CONTENTS

1.0 ABSTRACT ............................................................................................................................... 4

2.0 INTRODUCTION..................................................................................................................... 5

3.0 THEORY ................................................................................................................................... 7

4.0 METHODOLOGY ................................................................................................................. 13

5.0 RESULTS ................................................................................................................................ 14

6.0 DISCUSSION .......................................................................................................................... 20

7.0 CONCLUSION ....................................................................................................................... 25

8.0 RECOMMENDATION .......................................................................................................... 25

9.0 REFERENCES ........................................................................................................................ 26

3
 1.0 ABSTRACT

The objective for this experiment is to identify the unknown compound by using Ultraviolet-
visible Spectrometer (UV-Vis) and Infrared Spectrometer (IR). UV-vis is based on the
absorption of the ultraviolet light and visible light by chemical compounds which results in
the production of distinct spectra while infrared spectrometer is used to determine the
functional group of the unknown compound. The identification of the unknown compounds is
discussed through the spectra exhibited from both UV-Vis and IR. The unknown compounds
are identified by using the plotted logarithm epsilon against wavelength graph from the UV-
Vis result and the by plotting transmittance in percentage unit against wavelength of the
compound graph from the obtained IR results. Several peaks in the IR data are observed at
different intensities to determine the functional group. From all these data, the functional
group, the specific molecules in the compound and the structure of the unknown compound
are determined. From all the obtained data from UV-Vis and IR, the data are inserted in the
equation given until the equation are balanced. It can be concluded that the compound A, C,
D, I, J and K are identified as ethanol, ethanoic acid, ethyl ethanoate, benzoic acid, ethyl
bromide and diethyl ether, respectively.

4
 2.0 INTRODUCTION

The purpose for this experiment was to identify the organic compound by using Ultraviolet-
visible Spectroscopy (UV-Vis) and Infrared Spectroscopy (IR). Laboratory spectrometers are
used to identify the specific wavelength at which those compounds absorb visible light. These
specific information from the result can be used to identify what substance in the compound.

Ultraviolet and visible absorption spectroscopy is a type of spectroscopy which involved the
t passes through a sample or reflects from a
sample surface. UV-vis is based on the absorption of the ultraviolet light and visible light by
chemical compounds which results in the production of distinct spectra. When matter absorbs
ultraviolet radiation, the electrons present in it undergo excitation. This causes them to jump
from a ground state at which to an excited state at which an energy state with a relatively
large amount of energy associated with it. It is important to note that the difference in the
energies of the ground state and the excited state of the electron is always equal to the amount
of ultraviolet radiation or visible radiation absorbed by it [1].

The next spectrometer that is used in this experiment is infrared spectrometer. The infrared
spectrometer is used to determine the functional group of the unknown compound. This is an
important step because that information is used to determine the molecular weight of the
substance and the structure of the compound. The IR spectra is normally scanned from 4000-
400 cm-1. The first region which is between 4000-1500 cm-1 is called functional group
region. The peaks in this region are characteristic of specific kinds of bonds and it is used to
identify whether a specific functional group is present. The other region which is from 1500-
400 cm-1 is called fingerprint region. The peaks in this region arise from complex
deformations of the molecule. They may be characteristic of molecular symmetry, or
combination bands arising from multiple bonds deforming simultaneously [2]. Functional
groups have characteristic absorption peaks. These peaks are identified by their frequency
(energy) in wavenumbers and intensity of the absorption whether it is weak, medium or
strong. Covalent bonds absorb infrared radiation differently because it is depending on the
mass of the atom and strength and polarity of the bonds involved. Thus, running infrared
radiation on the unknown compound and measuring the absorbance at different wavelength
sheds light on the various bonds between atoms within the compound [3]. By gathering all

5
the information from the UV-vis and Infrared spectrometer, the molecular formula and
structure of al the unknown compounds are identified.

6
 3.0 THEORY

Spectroscopy is a common term used for instrumental methods from which by detailed study
of the absorption, dispersion or emission of electromagnetic radiation through molecules,
knowledge about molecular structure is collected. For example, visible light, the spectrum of
electromagnetic radiation detectable by human eyes, with wavelengths of about 380 to 780
nm. Spectroscopy in the ultraviolet and visible regions of the spectrum is referred to as
electronic spectroscopy, as it involves excitations of electronic states [4]. Electrons in an
atom or molecule are excited when the frequency of the incident radiation is equal to the
difference between the two electronic states in the energy source. The energy difference is
determined by the electronic structure of the molecules; the lower-energy proton is thus
necessary for vibrational shifts. The absorption spectrum is considered a plot between
absorption and the wavelength (nm) of incident radiation. The Beer-Lambert law shown as:
[1]
where, A is the absorbance (= log ( )),

coefficient, l is the sample length and c is the concentration. Equation [1] is formed by the
relationship between Lambert Law which show that the absorption of light by a sample
increases as the amount of absorbing material through which light has to travel increases, that
is the pathlength, l, increases (Figure 1) [5].

Figure 1: The relationship of the Beer-Lambert Law

If the pathlength increases, this reliance on pathlength is basically due to an increased number
of absorbing molecules in the light path (Figure 2).

7
 Figure 2: The relationship between the transmittance %
and path length

It is the basis of quantitative analysis using UV-Vis. The mechanism of Beer Lambert law
depends on beam of monochromatic parallel light that exposed to the the surface of the tested
medium. The medium absorbs part of the light energy after traveling through a medium of a
certain thickness, weakening the intensity of the emitted light passing through the medium
(Figure 3). The absorption of the absorbing medium is directly proportional to its thickness
[6].

Figure 3: Mechanism of Beer Lambert Law

A brief overview of how organic molecules interact with electro-magnetic radiation is needed
when dealing with UV-vis spectroscopy applications. A molecule is excited from the ground
state to an excited state when electromagnetic radiation is absorbed [7]. This refers to the
excitation of an electron from an occupied molecular orbital to an unoccupied (or singularly
occupied) molecular orbital for electromagnetic radiation within the UV-vis spectrum. This
coincides with the excitation of an electron from an occupied molecular orbital to an
unoccupied (or singularly occupied) molecular orbital for electromagnetic radiation in the
UV-vis range. The molecule can absorb light only if the energy of the light photons
corresponds to the energy difference between the two orbitals involved in the transition.
Shown as:

8
 [2]

(in Js), and n is frequency of the light (in hertz). Since it is better to visualize light in terms of

[3]

where c is the speed of light (in ). By combining those equations, a relationship

between the energy between two molecular orbitals and the wavelength of light required for
an elec- tronic transition between these orbitals can be shown as:

[4]

Thus, a shorter wavelength of light is needed to be absorbed for a larger energy gap between
two orbitals. For this cause, light absorption in the UV or visible region usually correlates
with lower-energy transitions involving valence electrons from non-bonded orbitals (i.e., lone
pairs) or p orbitals [8].
There are three selection rules to operate: 1) an electron must not change its spin quantum
number during a transition, 2) only one electron may be excited at a time. and 3) any
symmetry must be conserved during a transition. The main problem FOR UV-Vis is that an
analyte must have chromophore which is the function group [5]. Chromophores can be
classified into three different orbital environment which are 1) s-bonds, 2) lone-pair electrons,
and 3) p-bonds. Table 1 shows, transitions involving electrons originating in s-bonds usually
consume less than 200 nm and are thus of restricted usage for structure determination. Due to
the large energy difference between the bonding and antibonding orbitals, electronic
transitions involving s-bonds usually involve higher energy. As higher energy corresponds to

compounds, including oxygen and water, are absorbed in this region, it is important to obtain
spectra under vacuum conditions to access spectral information below 190 nm in the region.
In comparison, in non-bonded orbitals or p-orbitals, there are lower energy differences
between electrons and antibonding orbitals that occur in electronic transitions above 200 nm.
Compounds that have a high degree of conjugation will absorb at longer wavelengths and are
usually colored [8].

9
 Table 1: Typical Absorbance and Intensity of Chromophores

When the structure of the molecule is identified, that is, determined through the combined use
of other techniques such as infrared (IR) spectroscopy, the detection of a chromophore in a
molecule is readily achievable. Infrared (IR) spectroscopy, is the spectroscopy branch that
deals with the electromagnetic spectrum's infrared region, which is light with a longer
wavelength and a lower frequency than visible light [9]. It encompasses a variety of
techniques, often focused on spectroscopy of absorption. It can be used, as with other
spectroscopic methods, to classify and study chemicals. The Fourier Transform Infrared
(FTIR) spectrometer is a popular laboratory instrument that uses this technique. The IR is
divided into three different region which are near, mid and far with wavelengths between 3 x
10 4 and 3 x 10 3 cm [10].

Figure 4: IR Regions

10
There are two types of vibration which are bending and stretching [9]. In particular areas, the
vibrational frequencies of a given chemical group are supposed to depend on the type of
atoms involved and the type of chemical bonds. The frequencies of the chemical groups
within these vibration regions are modulated by the group's specific environment. Therefore,
it is appropriate to conduct a comprehensive IR analysis on condensed model compounds
indifferent conditions (solvents) and/or to evaluate experimental findings using theoretical
chemistry approaches to create a direct relationship between the infrared mode frequency and
the structural properties of a given residue, or if no data are available in the tables [11].
The interferogram is transformed into IR absorption spectrum that is usually recognize with
absorption intensity or % transmittance plotted against the wavelength or wavenumber [12].
The ratio of radiant power transmitted by the sample (I) relative to the radiant power of
incident light on the sample ( ) results in quantity of Transmittance, (T). Absorbance (A) is
the logarithm to the base 10 of the reciprocal of the transmittance (T) that are used to
determine organic compound in a solution. Different transmittance contributes to different
compound such as alcohols, carboxylic acids, aldehydes, ketones and esters [13].
[5]

Figure 5: FTIR spectrum

Based on previous research by using IR spectroscopy, Venelin Enchev calculated the heats of
formation, dipole moments, polarizabilities and ionization potentials of 96 compounds, eight

11
theoretically possible tautomeric forms of N-unsubstituted pyrazolones (hydroxypyrazoles)
and 11 of their C-substituted derivatives, by means of the MNDO method, with and without
configuration interaction (CI). The MNDO+ CI results for the relative stabilities are in
agreement with the available experimental data [14]. Suman Malik et.al have been
synthesized a Schiff base of 5-acetamido-1,3,4-thiadiazole--2-sulphonamide, using a
bidentate ligand, complexes of transition metals having the general formula ML2, where M =
Mn (II), Fe (II), Ni (II) and Cu (II), were synthesized. The complexes were characterized by
elemental analysis, molar conductivity, magnetic moment, electronic, ESR and IR
spectroscopy revealed an octahedral geometry for all the complexes [15].

12
 4.0 METHODOLOGY

UV-Vis Spectroscopy
10 mL square cuvette bottles were used to fill in a blank sample and organic compound
samples to be placed into the UV-Vis Spectrometer slots. 5 mL volumetric pipette was used
to measure the organic compound labelled as C and I and put into a 20mL volumetric flask
with lid. A Shimadzu UV-Vis 1800 spectrophotometer was used to evaluate the wavelength
of each sample. The resultant spectra of the organic compound samples were recorded using
the UV-Vis Spectrometer.

Infrared Spectroscopy
Samples of organic compound were prepared and labelled as A, C, D, I, J and K. The sample
holder disc was cleaned with acetone then an organic compound sample was dropped onto it
using a dropper. Then, the disc was firmly mounted inside the holder with the screw. The
vibrational bands for the organic compound samples were recorded using the Infrared (IR)
Spectroscopy.

Safety measures
Students need to read all procedures and associated safety information prior to the start of an
experiment. Dress for work in the laboratory by wearing cloth and shoes that cover exposed
skin and protect you from potential splashes. Also, practice good personal hygiene by
washing hands after removing gloves, before leaving the laboratory, and after handling a
potentially hazardous material.

13
 5.0 RESULTS

Compound A

Figure 6: IR spectrum of compound A

14
Compound C

Figure 7: UV-Vis spectrum of compound C

Figure 8: IR spectrum of compound C

15
Compound D

Figure 9: IR spectrum of compound D

16
Compound I

Figure 10: UV-Vis spectrum of compound I

Figure 11: IR spectrum of compound I

17
Compound J

Figure 12: IR spectrum of compound J

Compound K

Figure 13: IR spectrum of compound K

18
 Table 2: Characterization of Unknown Compound A, C, D, I, J and K

BAND (cm-1) BOND GROUP COMPOUND

Compound A

3321.09 O-H stretching (broad)

2942.79 -C-H stretching

1023.84 C-O stretching (Alcohol)

(Ethanol)
Compound C

3038.96 =C-H stretching

2629.96 O-H stretching

1757.00 C=O stretching

(Carboxylic Acid)
(Ethanoic Acid)
~1700.00 C=O stretching

Compound D

2984.60 -C-H stretching

1736.53 C=O stretching (Ester) (Ethyl Ethanoate)

Compound I

3323.72 O-H stretching (broad)

2976.76 -C-H stretching

1770.48 C=O stretching

(Carboxylic Acid) (Benzoic Acid)
1087.15 C-H bending

Compound J

2927.50 -C-H stretching

2990.31 -C-H stretching

(Alkyl Halide)
564.44 C-Br stretching (Ethyl Bromide)

Compound K

2977.22 -C-H stretching

2944.48 -C-H stretching

1076.15 C-O stretching (Ether) (Diethyl Ether)

19
 6.0 DISCUSSION

Several samples of organic compounds A, C, D, I, J, K had been collected for analysis using
UV-Vis and IR Spectroscopy for identification. The identification of the unknown
compounds is discussed through the spectra exhibited from both UV-Vis and IR [16].

In the infrared spectrum to identify unknown compound A, several peaks were observed at
different intensities. The functional group region showed a broad band at 3321 cm-1 which
could be attributed to the O-H group. The stretching at 2942.79 cm-1 and 2832.05 cm-1
suggested the appearance of -C-H group which indicated that only single bond alkyl is
present. The functional group region proposed that the compound is an alcohol. However, it
did not point to a specific alcohol compound, which led towards the fingerprint region for
final identification. In the fingerprint region, the assignment of the 1023.84 cm-1 band was
favourable to C-O stretching whose strong intensity is enhanced by the presence of primary
alcohol. This has led towards the identification of the alcohol compound to be ethanol.

In a study by Deepali L. Jadhav [17], an FTIR analysis had also been conducted on pure
ethanol as illustrated in the spectrum (a) below. The resultant spectrum showed similar
vibrational activity to the one obtained in unknown compound A. In the spectrum of pure
ethanol (a), the bands were assigned to the v(O-H), vs(CH3), vs(CH3), vas(CH2), vs(CH2),
v(C-O), and v(C-C-O), respectively. Similarly in a study by Elangovan et al. [18], the peaks
were almost identical.
Another unknown compound (compound C) was analysed using IR spectroscopy and UV-Vis
Spectroscopy. To identify this compound, IR spectroscopy was used in which a band at
3038.96 cm-1 can be observed that indicated the presence of alkenyl =C-H stretch. The
presence of carbonyl group C=O can also be observed at ~1700 cm-1 band. An O-H stretching
was also identified at the band of 2629.96 cm=1. These peaks have led us to conclude that the
compound belongs to the carboxylic acid group. To distinguish between aromatic and
aliphatic carboxylic acid compounds, there are slight differences which require further
analysis. One is the absence of overtones around the ~1700 cm -1 band. Another method is
through UV-Vis spectrum. It can be observed in the spectrum the maximum wavelength of
the compound is at 210 nm with a molar extinction of 32. This wavelength could indicate the
presence of aliphatic -COOH group. Therefore, this analysis has led to the identification of
ethanoic acid. In a work by Chandraiah [19], an FTIR study had been made in ethanoic acid

20
mixtures. Similarly to the results obtained in unknown compound C, a broad O-H band could
also be observed at 3040 cm=1. A sharp band at 1070 cm=1 also made an appearance which
indicated the stretching of C=O frequency of ethanoic acid.

Unknown compound D was also analysed using IR spectroscopy. The 2984.60 cm-1 band was
determined to be the alkyl C-H stretch which are fairly ubiquitous that make it less useful in
determining the structure. However, a strong band at 1736.53 cm=1 could also be analysed in
the compound that favour towards the presence of ester (-COOR) group. This identification is
further amplified by the strong peak at ~1200 cm=1 which represents ester C-O stretching.
Therefore, this compound is concluded to be ethyl ethanoate.

Li, F. in their work on GC MS, FTIR and Raman analysis of antioxidant components of red
pigments from Stemphylium lycopersici [20] also demonstrated similar infrared spectrum as
unknown compound D. A broad stretching peak was observed at 3414 cm-1 which was
attributed to the O-H vibrational activity. The presence of the O-H stretching (which is absent
in compound D) might be a result of other compound in the ethyl acetate extraction from
Stemphylium lycopersici. Similar to unknown compound D, a stretching peak could also be
seen at around 1650 cm=1 which favoured the C=O presence. An attribution of C-O was also
evident at 1205 and 1160 cm=1 which favoured an ester group compound.

IR spectroscopy was also used to analyse unknown compound I which yielded in IR spectrum
of a number of peaks. The most notable bands in the functional group region are the ones at
3323.72 cm-1 and ~1700 cm-1. The former indicates towards the appearance of the broad O-H
stretching meanwhile the latter proposes that the C=O stretching of monomer carboxylic acid
may inhabit the compound. The in-plane aromatic C H bending vibration occurs in the
region of 1142.49 and 1087.15 cm=1 considering the bands are sharp but have weak-to
medium intensity. The slight overtones in the functional group region also suggests an
aromatic compound. To further validate this inference, UV-Vis spectroscopy was used. In the
spectrum generated, it can be interpreted that the compound has a maximum wavelength of
220 nm with a molar extinction of ~2000. This implies the absorbance of the UV light by an
aromatic compound. Therefore, it is hypothesized that compound I is benzoic acid. In a study
by C Meganathan [21], several vibrational bands could be noticed to indicate a benzoic acid
compound. A broad band appearing at 3354 cm-1 and 1765 cm-1 was characterized by the
presence of O-H and C=O stretching, respectively. Saad, B. [22] also illustrated a UV

21
spectrum of benzoic acid in his study. Based on figure above, it is observed that the
maximum wavelength of benzoic acid was approximately 230 nm which almost resembled
the UV spectrum result of compound I.

The functional group region in the IR spectrum of unknown compound J resulted in observed
band at ~2900 cm=1 which represent the alkyl C-H stretch. It is rather difficult to identify the
compound given only the C-H stretch as the functional group. However, overlapping
vibration of ~564 cm=1 could be seen in the fingerprint region which can be inferred to be a
Br compound. Therefore, the compound is likely to be an alkyl halide (R-X) of ethyl
bromide. Arlinghaus, R.T. [23] studied on the FTIR spectra of alkyl halides. In the spectrum,
there was a band observed at 603 cm=1 which was attributed to the fundamental bonding of
C-Br of methyl bromide. This spectrum further adds to the identification of compound J to be
an alkyl bromide.

The IR spectrum of unknown compound K had yielded multiple peaks of ~2900 cm -1 which
also represent the alkyl C-H stretch. The fairly less useful indicator had also led us towards
analysing the fingerprint region as well. A strong stretching at ~1100 cm-1 band could be
notably be seen in the spectrum in which the peak favoured the presence of C-O bond of
aliphatic ether [24]. Therefore, it is deduced that compound K is an ether, and more precisely,
the diethyl ether.

For compound B the IR spectrum peak is 1730 cm-1 will represent the C=O bond. Due to the
wide options on specific compound B as it has single peak the UV-vis reading need to be

compound is ethanal. Unknown compound E can be determined as the peak 2950 and 1685
cm-1 show that the compound have alkyl C-H stretch and alkenyl C=O stretch. From the

carboxyl or amino group as it absorbs light below 220nm and at wavelength of 273 nm with
-COOH functional group. It can be said that the unknown E is phthalic
acid as there are two carboxylic acid attach to the benzene.

Based on the peaks of IR spectrum for unknown compound F at 3430cm-1 there is N-H
stretch contain in the compound. At peak 1592cm-1 it can be said that unknown F carry C=C

22
bond in the compound. The maximum wavelength for the UV-
>3.2 also show appearance of N-H bond in the unknown compound F. The possible
functional group for the compound F is diamino ethylene.

IR spectrum for compound G have peak at 3037cm -1 represent the compound have alkyne C-
H stretching and peak at 1480 cm-1 show that CH2 bend with medium intensity is included in
the compound. The spectrum that has maximum wavelength of 178 nm by the reading can be
said that C=C bond exist. From the data, the compound G can be concluded as benzene.

Unknown compound H can be determined as toluene as there is a few peaks for IR spectrum
and UV-vis reading that prove it. The first peak at 3033cm-1 show that the compound carry
C-H stretching and other peak at 1501cm-1 give a meaning that it has aromatic C=C bending
in unknown H. Plus, it also has aromatic C-H bending as there is a peak of IR spectroscopy at
730cm-1. For the UV-
group.

For further confirmation of the compounds identified, few equations were made to
completely know the compounds involved in this experiment.

Through the equation when compound A that is ethanol react with reagent pyridinium
chlorochromate (PCC) with present of dichloromethane the result will get compound B that is
acetaldehyde. Due to oxidising agent PCC the primary alcohol ethanol will turn into ethanal
as the reaction is known as oxidation of alcohol [25]. Usually, the alcohol will be oxidized to
carboxylic acid when in aqueous environment but in this experiment the uses of PCC and

23
dichloromethane as solvent rather than water make the oxidation stop at aldehyde stage and
produce the acetaldehyde as product. With further experiment the compound B (aldehyde
group) react with Chromium(III) with present of sulfuric acid. The Chromium(III) act as
strong oxidising agent that not soluble in most organic solvent that why the sulfuric acid is
use as solvent as it is safe to react with the aldehyde to produce carboxylic acid [26]. The
oxidation of acetaldehyde will produce ethanoic acid and it prove that the IR spectroscopy
reading for compound C is correct.

In second equation, compound A (ethanol) react with compound C (ethanoic acid) with the
existing of HCl will produce compound D. When a carboxylic acid is treated with an alcohol
with the present of acid catalyst, an ester and water will be produced. From the explanation
the compound D, it is likely to be ethyl ethanoate as the ethanol react with ethanoic acid with
the present of HCl.

As the compound G is identified as benzene, in the equation the compound react with
chloromethane with present of FeCl3 will produce compound H. The CH3Cl in the presence
of FeCl3 act as alkylation agent and the reaction known as Friedal-
benzene [27]. Through the reaction toluene and HCl will be formed as the CH3 from CH3Cl
will attach to the benzene and one H+ from benzene will attach to Cl- to form HCl. The
compound H (toluene) next been react with KMnO4 with water to produce compound I. The
KMnO4 present as oxidising agent in this reaction and when the toluene is being oxidised it
will produce benzoic acid.

The compound K is produced when ethanol reacts with bromoethane with present of NaH
and THF. The NaH purpose isto deprotonate the ethanol (forming H2 in the process), making
it into a nucleophilic alkoxide ion, which then performs a substitution reaction [SN2
mechanism][28]. From the reaction ester is produced as diethyl ether.

24
 7.0 CONCLUSION

In conclusion, the objectives for this experiment were successfully achieved. This experiment
was conducted to identify the unknown compound by using Ultraviolet-Visible (UV-Vis)
Spectrometer and Infrared (IR) Spectrometer. Spectrometer was used in this experiment to
measure the transmittance and absorption of each compound given. UV-Vis spectrometer and
IR spectrometer were applied for this experiment to identify the compounds given by using
ultraviolet light and infrared light, respectively. The samples of organic compounds, A, C, D,
I, J, K were identified through the spectra resulted from both UV-Vis and IR. Compound B,
E, F, G, H were identified for this experiment from the chemical reaction given as a guidance
to determine the compound of A, C, D, I, J, K. It shows that compound A is identified as
ethanol due to the results represents the functional group of alcohol and as a primary alcohol
for final identification from the fingerprint region. Compound C and I is ethanoic acid and
benzoic acid respectively as the carboxylic acid group were absence in these compounds.
Functional group of esters is identified in compound D is determined as ethyl ethanoate.
Next, compound J is discovered as ethyl bromide due to the presence of alkyls halide inside
this compound. Lastly, compound K is found to be an ether group and can be labelled as
diethyl ether. At the end of experiment, all 11 compounds (A-K) were successfully identified
by using the knowledge of UV-Vis and IR spectroscopy. Furthermore, naming and justifying
every compound by using the information given were successfully identified.

8.0 RECOMMENDATION

To ensure the experiment conducted smoothly, the students should be well prepared before
conducting the experiment without facing confusion about the steps to be done during the
experiment. Next, students should learn properly how to handle the spectrophotometer as the
cuvette bottles is very sensitive. Failing to do this will influence the experiment for getting
less accurate and precision of absorbance measurements and should be carefully controlled.
Furthermore, the cleanliness of the cuvettes is very important as it has major effects on
results. Fingerprints and grease that are deposited on the cuvette windows will affects the
absorbance data. Thus, cleaning thoroughly before and after needs to be done to prevent these
matters from happening. Lastly, the eyes should be parallel to the level measurement scale of
pipette to achieve accurate reading and data, and also to avoid human error in this
experiment.

25
 9.0 REFERENCES

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[3] Kobunshi, vol. 23, no. 5, pp. 378-383,388,

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[4] H. Günzler and A. Williams, Handbook of Analytical Techniques, vol. 1 2. 2008.

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[8] A. A. Edwards and B. D. Alexander, UV-visible absorption spectroscopy, organic

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[17] Jadhav, D. L., Karthick, N. K., Kannan, P. P., Shanmugam, R., Elangovan, A., &
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