UV-VIS Experiment AS230

UNIVERSITY TEKNOLOGI MARA
FACULTY OF APPLIED SCIENCES
MST516
LABORATORY REPORT
ULTRAVIOLET-VISIBLE SPECTROSCOPY
NAME : QAMARUL IZZAT BIN SHAHROM
ID : 2016647236
GROUP : AS2303M2
PREPARED FOR : DR MOHD MUZAMIR BIN MAHAT

EXPERIMENT 1
TITLE
ULTRAVIOLET VISIBLE (UV-VIS) SPECTROSCOPY
OBJECTIVES
1. To determine wavelength of maximum absorption, max of the solution.
2. To determine absorbance of solution of different concentration of curcumin at the

wavelength of maximum absorption.
3. To determine the concentration of unknown solution.
INTRODUCTION
UV-VIS spectroscopy is a very functioning device in order to achieve many

objectives from detecting functional group in a materials or detection of impurities
and so many more. Theoretically, UV electromagnetic radiation can range from
200-400nm and for Visible can range from 200-700nm. Sometimes, visible
electromagnetic radiation can exceed a bit from 700nm. In this introduction, a very
typical instrumentation set for spectrometer of UV-VIS are shown below. It is quite
straightforward. The light source that emits the light whether UV or visible light will
pass through the first slit and the separated into its components wavelength by the
diffraction grating or prism. After that, the separated beam that becomes
monochromatic single wavelength will be split into two intensity beams by
half-mirror. One of the split beam will pass through a small transparent cuvette
containing the sample solution. This beam will be regarded as sample beam. The other
beam, will act as reference beam that pass through the same cuvette containing the
reference solvent. All these two intensities of light will be detected by the electronic
detectors. The detection of these beams by the electronic detectors will be compared.
The intensity of reference beam will experience little or no absorption, defined as Io.
Meanwhile, the intensity of the sample beam is defined as I.
MATERIALS
Curcumin Stock solution of 100 ppm, one unknown curcumin solution and methanol
Glassware : Six 10 ml volumetric flask, two 10 ml pipets, and one 150 ml beaker
PROCEDURE
1) 10 ml each of 5, 10, 15, 20, and 25 ppm solution of curcumin is prepared from
stock solution of 100 ppm.*
*Use dilution formula :
M1V1 = M2V2
Where M1 = Concentration of stock solution
V1 = Volume required from stock solution
M2 = Concentration of working solution
V2 = Volume of volumetric flask

Measurement of samples using PERKIN ELMER LAMBDA 35 Spectrophotometer :
2) All the instrument and computer are switched on.
3) Icon LAMBDA 35 is double clicked and wait until all the process (initializing).
4) Scan icon is clicked to scan the spectrum within the specified range wavelength
by keying in the values of the wavelength to start and the wavelength to end.
5) Inst icon is clicked to state whether the absorbance (A) value or transmission
(%T) value will be measured.
6) Sample icon is clicked to state the result filename, the number of sample, and
named the sample identify in arrangement flow.
7) After that the start icon is clicked.
8) Wait until please insert your name sample appear then sample is inserted into
the cell compartment of the instrument that is hearer without taking out the
reference sample. The compartment is closed and OK is clicked.
9) The step is repeated for the next sample.
10) After finishing all the scanning, x-axis and y-axis is clicked to change the formal
of abscissor range (vertical) and ordinate range (horizontal).
11) Peak Icon is clicked to get the automatic peak label; peak choosed display both.
12) V-cursor is clicked to find other peaks and make sure that v-cursor is on the peak.
13) Then, V-cursor is double-clicked to get another peak and the step was repeated
for another peak labelling.
14) After finishing, all the peaks are labelled. ABC is click then sample of the
experiment is named and title experiment is clicked OK.
15) Our sample is printed directly or the file is clicked and copy option is chosen to
clipboard. Then pasted to word file Microsoft Word.
RESULTS AND DISCUSSION
Dilution calculation by using formula:
M1V1 = M2V2
Concentration of working solution,M2 Value of Volume required from stock

solution, V1 (ml)
5 100 (V1) = 5 (10)

100 (V1) = 50
V1 = 0.5
10 100 (V1) = 10 (10)

100 (V1) = 100
V1 = 1.0
15 100 (V1) = 15 (10)

100 (V1) = 150
V1 = 1.5
20 100 (V1) = 20 (10)

100 (V1) = 200
V1 = 2.0
Concentration of solution (ppm) Absorbance (A)

5 0.07
10 0.21
15 0.14
20 0.27
25 0.34
Figure 2: The graph of concentration of solution versus absorbance.

Figure 3: Spectrum Graph
Graph of Absorbance vs Concentration

0.4
0.35
0.3
Absorbance (A)
0.25
0.2
0.15
0.1
0.05
0
0 5 10 15 20 25
Concentration (ppm)
Figure 4: Graph of Absorbance vs Concentration

In this experiment, we prepared 5 solution with different concentration to
determine their maximum absorption. We had plotted a graph of concentration of
solution versus absorbance. We can see that concentration of solution is directly
propotional to absorbance. The absorbance increase when the concentration of the
solution is increased. As we look at the Figure 3, we can say that the wavelength of
maximum absorption is 525.42 at the absorbance of
As from the results we achieved, we can consider that the experiment that we did
is quite a successful experiment. This is because we can show the relation of
concentration and absorbance that is directly proportional. Practically, we achieved
the results as expected from the theoretically. But all human make mistakes, and so
did us. At the concentration of 15 ppm, we have undergone some clumsiness in the
preparation of the solution. Based on Figure 4, it can be seen how the curve of the
graph went down on the 15 ppm.
This happens because of several errors of our own. Firstly, we did not really
well-prepared before doing the experiment. We did not study the experiment really
well and this leads to the error. Secondly, while handling the experiment we were too
clumsy about the preparation of the solution. We have done errors especially at the
time where our eyes must be sharp at the lever knob, the usage of the dropper, and the
handling of the cuvettes. We must follow the instruction from the manual lab how to
handle that instrument well.
CONCLUSION
From this experiment, we know how to use the instrument and its function. We
also can determine the wavelength of maximum absorption, max of the solution
and absorbance of solution of different concentration of curcumin at the wavelength
of maximum absorption. we can conclude that this experiment is successful because
we had achieve all the objectives given for this experiment.
REFERENCES
1. William, R. (2013). Visible and Ultraviolet Spectroscopy. Retrieved October 29,

2017, from
http://www2.chemistry.msu.edu/faculty/reusch/virttxtjml/spectrpy/uv-vis/spectrum.ht
2. Michigan State University, Department of Chemistry. Retrieved 29, 2017, from

http://www2.chemistry.msu.edu/faculty/reusch/virttxtjml/spectrpy/uv-vis/uvspec.htm#
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UNIVERSITY TEKNOLOGI MARA

FACULTY OF APPLIED SCIENCES

NAME : QAMARUL IZZAT BIN SHAHROM

PREPARED FOR : DR MOHD MUZAMIR BIN MAHAT

ULTRAVIOLET VISIBLE (UV-VIS) SPECTROSCOPY

1. To determine wavelength of maximum absorption, max of the solution.

2. To determine absorbance of solution of different concentration of curcumin at the

3. To determine the concentration of unknown solution.

UV-VIS spectroscopy is a very functioning device in order to achieve many

*Use dilution formula :

Where M1 = Concentration of stock solution

V1 = Volume required from stock solution

M2 = Concentration of working solution

V2 = Volume of volumetric flask

2) All the instrument and computer are switched on.

7) After that the start icon is clicked.

9) The step is repeated for the next sample.

RESULTS AND DISCUSSION

Dilution calculation by using formula:

Concentration of working solution,M2 Value of Volume required from stock

5 100 (V1) = 5 (10)

10 100 (V1) = 10 (10)

15 100 (V1) = 15 (10)

20 100 (V1) = 20 (10)

Concentration of solution (ppm) Absorbance (A)

Figure 2: The graph of concentration of solution versus absorbance.

Graph of Absorbance vs Concentration

Figure 4: Graph of Absorbance vs Concentration

1. William, R. (2013). Visible and Ultraviolet Spectroscopy. Retrieved October 29,

2. Michigan State University, Department of Chemistry. Retrieved 29, 2017, from

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