FACULTY OF APPLIED SCIENCES

CHM 571 - BASIC INSTRUMENTAL ANALYSIS

LAB REPORT

NAME: MOHAMAD SAIFUL BIN MOHD RAFFIAH

STUDENT ID: 2021459156
CLASS GROUP: AS239 4A
EXPERIMENT 5: ULTRA VIOLET / VISIBLE SPECTROSCOPY
LECTURER: DR. NORAINI BINTI KASIM
DATE OF EXPERIMENT: 24th MAY 2023
DATE OF SUBMISSION: 02nd JULY 2023
Tittle : Ultra Violet / Visible Spectroscopy

Objective
1. To scan and obtain the absorption spectra of KMnO4 solution (20 ppm and 40 ppm) at
wavelength range 350 to 700 nm.
2. To determine the wavelength of maximum absorbance (λ max) from the spectrum.
3. To produce a standard calibration plot from a series of standard solutions and calculate the
molar absorptivity using Beer’s law.
4. To determine the concentration of KMnO4 solution.

Calculation In Pre-Lab Question

KMnO4 solution in 50 ml volumetric flask from standard solution (100 ppm)
Concentration (ppm) Volume (mL)
10 500
20 250
30 125
40 100

100 ppm KMnO4 solution is prepared from the stock solution

A series of standard solutions from the above stock solution is prepared to a concentration of 5,
10, 15, 20, 25, 30, 35 and 40 ppm.

10 ppm
M1V1 = M2V2
100 ppm(50) = 10 V2
V2 = 500ml

20ppm
M1V1 = M2V2
100 ppm(50) = 20 V2
V2 = 250ml

40ppm
M1V1 = M2V2
100 ppm(50) = 40 V2
V2 = 125ml

50ppm
M1V1 = M2V2
100 ppm(50) = 50 V2
V2 = 100ml

Introduction
UV-Vis spectroscopy is an analytical technique that compares the amount of discrete wavelengths
of UV or visible light absorbed or transmitted by a sample to a reference or blank sample. The
sample composition impacts the characteristic, which may provide information about what is in
the sample and their concentration. Considering this spectroscopy approach is based on the
usage of light, we should first look at the properties of light.

Light contains energy that is inversely proportionate to its wavelength. As a result, shorter
wavelengths of light carry more energy, whereas longer wavelengths carry less. A certain amount
of energy is required to promote electrons in a substance to a higher energy state, which can
detect as absorption. Electrons in various bonding sites in a substance require a variable amount
of energy to be promoted to a higher energy state. This is why light absorbs at different
wavelengths in different substances. Humans can see a spectrum of visible light ranging from
around 380 nm as violet and up to 780 nm as red. UV light has wavelengths that are shorter than
visible light up to 100 nm. As a result, light may be defined by its wavelength, which can be used
in UV-Vis spectroscopy to analyse or identify different substances by locating the exact
wavelengths corresponding to maximum absorbance.

The Perkin Elmer Lambda 25 UV/VIS spectrometer was used for this experiment. At first, 100 pm
KMnO4 was made from a stock solution of 1000 pm KMnO4. We created 100 pm KMnO4 and
diluted it into 5, 10, 15, 20, 25, 30, 35, 40ppm and unknown. To adjust the spectrophotometer, a
blank was run before inserting other concentrations to set the instrument to autozero. Following
that, absorption spectra of 20 and 40 pm in the 350-700 nm wavelength region were obtained.
The wavelength of maximum absorbance was determined from the data acquired as an outcome
of the absorption spectrum. The graph showed the absorbance calculated from the 5, 10, 15, 20,
25, 30, 35, and 40 ppm. The Least Squares Method was used to determine the best fit line graph.
The graph can be used to determine the concentration of an unknown substance.
Materials and Apparatus
• Stock KMnO4 solution (1000 ppm)
• KMnO4 solution (unknown concentration)
• Perkin Elmer Lambda 25 UV/VIS spectrometer

Procedures
1. A 1000 ppm standard stock solution was used to make a 100 ppm KMnO4 solution. Formula
C1V1 = C2V2 was used to determine the KMnO4 volume to be pipetted. Transferring 25 ml of
KMnO4 solution into a 250 ml volumetric flask with a pipette.
2. The solution was then diluted with deionized water until it reached the calibration point. A
stopper was placed on the volumetric flask, and the solution was shaken to blend it. The
standard solutions were then produced at concentrations of 5, 10, 15, 20, 25, 30, 35, and 40
ppm.
3. Following that 5, 10, 15, 20, 25, 30, 35, and 40 ml of the 100 ppm KMnO4 solution were pipetted
into eight separate 100 ml volumetric flasks, one of which served as a blank solution. The
solution was diluted with distilled water until it reached the calibration point.
4. The volumetric flask was covered with a stopper, and the solution was thoroughly shaken to
combine it. To eliminate impurities before measuring the wavelength of KMnO4 with a Perkin
Elmer Lambda 25 UV/VIS spectrometer, a spectrometer cuvette was rinsed with distilled water.
5. After that, the KMnO4 solution was tested at 20 and 40 ppm. The KMnO4 solution was then
partially poured into the cuvette. Before gently inserting the cuvette into the spectrometer, the
external wall of the cuvette was wiped using the tissue paper.
6. The radiation was directed at the cuvette's transparent side. The spectrometer had been turned
on and the holder was shut. Later, 5 ppm, 10 ppm, 15 ppm, 20 ppm, 25 ppm, 30 ppm, 35 ppm,
and 40 ppm were tested.
7. Finally, for all measurements, the unknown sample was run in the same method as the
cuvettes. The results were printed once all of the samples had been analysed.
Data & Results

Calculation
Concentration, Absorbance Average
Standard
ppm 1 2 3 Absorbance
Blank 0 0.003 0.003 0.002 2.667
1 5 0.120 0.120 0.119 0.120
2 10 0.306 0.305 0.306 0.305
3 15 0.347 0.347 0.346 0.346
4 20 0.478 0.477 0.477 0.477
5 25 0.574 0.573 0.573 0.573
6 30 0.761 0.761 0.760 0.760
7 35 0.863 0.863 0.862 0.862
8 40 0.937 0.937 0.936 0.936
Unknown 30 0.583 0.583 0.583 0.583

Preparation of 100 ppm KMnO4 solution from 1000 ppm standard solution, calculate the required
volume using formula C1V1 = C2V2.

C1V1 = C2V2
(1000 ppm) V1 = (1000 ppm) (250 ml)
(1000 ppm) V1 = 25000 ml
V1 = 25 ml

Preparation of 5 ppm, 10 ppm, 15 ppm, 20 ppm, 25 ppm, 30 ppm, 35 ppm, and 40 ppm of KMnO4
solution in 100 mL volumetric flask using formula C1V1 = C2V2.

1. 5 ppm
C1V1 = C2V2
(100 ppm) V1 = (5 ppm) (100 ml)
V1 = 5 ml

2. 10 ppm
C1V1 = C2V2
(100 ppm) V1 = (10 ppm) (100 ml)
 V1 = 10 ml

3. 15 ppm
C1V1 = C2V2
(100 ppm) V1 = (15 ppm) (100 ml)
V1 = 15 ml

4. 20 ppm
C1V1 = C2V2
(100 ppm) V1 = (20 ppm) (100 ml)
V1 = 20 ml

5. 25 ppm
C1V1 = C2V2
(100 ppm) V1 = (25 ppm) (100 ml)
V1 = 25 ml

6. 30 ppm
C1V1 = C2V2
(100 ppm) V1 = (30 ppm) (100 ml)
V1 = 30 ml

7. 35 ppm
C1V1 = C2V2
(100 ppm) V1 = (35 ppm) (100 ml)
V1 = 35 ml

8. 40 ppm
C1V1 = C2V2
(100 ppm) V1 = (40 ppm) (100 ml)
V1 = 40 ml
Discussion

The result was acquired to determine how the concentration of potassium manganate KMnO4,
and absorbance changed over time. The experiment's outcome was determined by the absorption
spectra through the maximum wavelength. By injecting the data of the absorbance and plotting
on the graph, it is possible to determine the concentration of the solution using the formula from
the Beer Lambert law. The formula used to determine a sample's composition based on how
much light it absorbs using Beer's law A = εbc . A compound's molar absorptivity is used to gauge
how strongly it absorbs light at a specific wavelength. While c is for the sample concentration and
b is for the path length. The molecules pass from a low energy state to a higher energy state from
the radiation field as a result of energy absorption. The amount of absorbance will increase as
the standard solution is concentrated. The sample, which may go through modifications like
association or dissociation, and the solvent used to dissolve the sample both have the potential
to introduce error or sources of error that could impair the accuracy of the results. The incident
light's wavelength may span a large wavelength range and not be strictly monochromatic.
Additionally, inaccurate readout from detectors, sample preparation, wrong wavelength settings,
and careless handling of cells may result in the measurement of transmittance and absorbance.

Conclusion
To conclude, by consideration of the beer Lambert rule, the experiment highlights the use of
ultraviolet-visible spectroscopy to measure the absorbance of the chemical potassium
manganate, KMnO4. The concentration of the solution and the length of the solution are inversely
related to the amount of light absorbed by the solution prepared by 5, 10, 15, 20, 25, 30, 35, 40,
and unknown sample.

Reference

Students’ resource: UV / Visible spectroscopy. (n.d.). RSC Education.

https://edu.rsc.org/resources/students-resource-uv-/-visible-spectroscopy/282.article