BASIC INSTRUMENTAL ANALYSIS

EXPERIMENT 2: UV- Visible Determination of an Unknown

Concentration of KMnO4 Solution

PREPARED BY

NAME : SITI MAIZATUL AKMA BINTI USLEY

UITM ID NUMBER : 2018422676

CLASS/GROUP : AS120 5J

PREPARED FOR

MISS NUR SYUHADA BINTI ISMAIL

SUBMISSION DATE

18 NOVEMBER 2020
Introduction
All absorbance spectrophotometers contain a light source, a sample compartment, and a
detector. Many spectrophotometers also contain one or more monochromators, a device used
to separate light into its component wavelengths. Spectrophotometers that measure in the UV
and visible region are of two general types: scanning and diode-array. In contrast, a scanning
UV-Vis spectrophotometer contains a monochromator, usually consisting of holographic
gratings, which allows light of individual wavelength to be sequentially imparted to the
sample. Spectroscopy involves the observations of absorption or emission of electromagnetic
radiation resulting from transitions of atoms or molecules from one energy level to another.
When a molecule at its “ground state” (the state of lowest energy) absorbs energy of some
type, the molecule is said to undergo a “transition” to a higher energy state. The higher
energy state is referred to as an “excited state”. A molecule can only absorb energy if the
input energy exactly matches a molecular transition from one energy level to another. Many
organic and biological molecules have transitions that occur between energy levels of
electronic states of atoms or molecules. Therefore, our focus will be on the visible and
ultraviolet regions of the electromagnetic spectrum in this lab. The visible and ultraviolet
region of interest is found between 170 and 800 nm, though the most useful region for
experimental use is between 250 and 700 nm.

Procedure
A. Preparation of the KMnO4 standard solutions.
1. 0.01 g KMnO4 is weighed accurately to the nearest mg on a weighing paper. The
reading is recorded. The solid is transferred to a 100 ml volumetric flask by using a
funnel.
2. The solid is dissolved with a few ml of distilled water. The flask is stopped and shake.
The distilled water is added to the mark and the last few drops is added by using a
medicine dropper. The flask is stoppered and shaken several times to homogenize the
solution.
3. The ‘stock’ solution is poured into a beaker. The beaker is labelled as ‘100 ppm’.
4. 5 ml of the ‘stock’ solution is pipetted and diluted with distilled water in a 100 ml
volumetric flask.
5. The solution is transferred into a beaker and the beaker is labelled as ‘5 ppm’.
6. Step 4 repeated by using 10 ml and 15 ml stock solution and then are transferred into
small beakers.
7. The beakers labelled as ’10 ppm’ and ’15 ppm’ respectively.
B. Preparation of the unknown
1. Between 5 to 20 ml of the ‘stock’ KMnO4 solution pipetted and diluted with distilled
water in a 100 ml volumetric flask.
2. The solution transferred into a beaker and it labelled as ‘Unknown’.
C. Operation of the UV-Vis Spectrophotometer
Instrument: Varian / Cary 50 UV-Vis Spectrophotometer.
Operating instructions
1. Cary Win UV icon selected.
2. ‘Scan’ clicked.
3. Setup chosen, the ‘CARY’ is clicked and then the required start and
stop scan wavelength (nm) is keyed in.

I. Y-mode (min = 0 and max = 1)

II. X-mode = 800 -200 nm
III. Beam mode = Dual Beam

4. Cycle mode is selected if more than 1 cycle is required.

5. ‘Scan Control Speed’ is selected to fast.
6. ‘Baseline’ icon and check ‘Baseline Correction Function’ clicked.
7. ‘Accessories 1’ clicked and checked on the Use Cell Charger cells
selected.
8. At the peak table option, maximum peak selected.
9. In the Auto Store icon, ‘storage on (Prompt At Start)’ was set.
10. To save the method, go to the file and the scan method is saved.
11. The ‘BLANK’ cuvette solution is put and ‘Baseline’ is clicked.
12. The ‘BLANK’ cuvette solution is put and ‘Baseline’ is clicked.
13. The ‘BLANK’ cuvette is removed and the ‘SAMPLE’ cuvette is put.
14. ‘Start’ icon clicked to start the measurement.

D. Determination of the unknown concentration

1. The concentration icon clicked.
2. Setup is chosen, Cary icon is clicked, then maximum wavelength, λ max is
keyed in.
3. Replicate = 3 is selected.
4. ‘Standard’ icon is clicked, the ‘Calibrate During Run’ function is checked.
5. The calibration standard unit (mg/L) and the number of the standard samples
is set.
6. The Fit Type (Linear Direct) is selected.
 7. Sample icon is chosen, the number of the samples are selected and the
unknown is keyed in.
8. Report icon is clicked, operator name and the comment is keyed in.
9. In the Auto Store icon, ‘Storage on (Prompt at start)’ is clicked.
10. To save the method’, ‘File Save Method As Ok’ is clicked.
11. The ‘BLANK’ cuvette is put and ‘ Zero’ is clicked.
12. The ‘BLANK’ cuvette is removed and the ‘SAMPLE’ cuvette is put.
13. ‘Start’ icon is clicked to start the concentration measurements.

Data and calculation

Name and model of instrument: UV/1800 SHIMADZU UV SPECTROPHOTOMETER
Table 2.1: Table of concentration and absorbance

Solution Concentration (ppm) Absorbance

Standard 1 5 0.155

Standard 2 10 0.231

Standard 3 15 0.299

Unknown ? 0.262

Mass of KMnO4 = 0.01 g λ max = 525 nm

Show the sample calculation for the preparation of standard 3:

y = 0.0144x + 0.0843
x = 15 mL of KMnO4 ‘stock’ solution is used to prepare standard 3 solution
y = 0.0144(15) + 0.0843
= 0.300
 A Graph of Absorbance versus Concentration of KMnO4
0.35

0.3
f(x) = 0.04 x + 0.14
0.25 R² = 0.67
Absorbance

0.2

0.15

0.1

0.05

0
0 2 4 6 8 10 12 14 16 18 20
Concentration (ppm)

y = 0.0144x + 0.0843
y = absorbance = 0.262
x = concentration
0.262 = 0.0144X + 0.0843
X = (0.262 – 0.0843) / 0.0144
= 12.34
Then, concentration of unknown is 12.34 ppm

Discussion
The objectives of this experiment are to determine the maximum wavelength of potassium
permanganate, to plot the calibration curve of potassium permanganate and to determine the
concentration of an unknown solution of potassium permanganate. At first in this experiment,
an initial permanganate stock solution is prepared and the solutions to be measured that are 5
mL, 10 mL and 15 mL and also unknown solution are diluted from a dilution of the stock.
Once the absorbance value are taken, Beer’s law plot for KMnO4 is generated and the
concentration of unknown solution can be determined.
The instrument that we have used to do this experiment is UV-Vis Spectrophotometer. UV
Visible spectroscopy measures the response of a sample to ultraviolet and visible range of
electromagnetic radiations. Molecules and atoms have electronic transitions while most of the
solids have interband transitions in the UV and Visible range. The instrument used in
ultraviolet-visible spectroscopy is called a UV/VIS spectrophotometer. It measures the
intensity of light passing through a sample (I) and compares it to the intensity of light before
it passes through the sample (Io). The ratio I/Io is called the transmittance and usually is
expressed as a percentage (%T). The absorbance, A is based on the transmittance that is
A= - log T. The UV-visible spectrophotometer can also be configured to measure reflectance.
In this case, the spectrophotometer measures the intensity of light reflected from a sample (I)
and compares it to the intensity of light reflected from a reference material (Io) such as white
tile. The ratio I/I0 is called the reflectance and usually is expressed as a percentage %R).
Instruments for measuring the absorption of UV or visible radiation are made up of the
following components that are light source, spectroscope, sample containers and a detector.
Two kinds of lamps, a Deuterium for measurement in the ultraviolet range and a tungsten
lamp for measurement in the visible and near-infrared ranges are used as the light sources of
a spectrophotometer. In tungsten lamp, a continuous spectrum of 300-3000 nm is emitted
while in deuterium lamp, only a continuous spectrum of 168-500 nm, with maximum energy
at 250 nm is emitted. Other component in this instrument is spectroscope. A spectroscope
plays a role in selecting a monochromatic light from a light source (white light).
Spectroscope include Filter type, Prism type and Grating (diffraction grating) type. In filter
type, a single wavelength can be extracted with a filter. A filter is also used in combination
with diffraction grating for filtering out stray light. For prism type, a spectrum of 175-2700
nm can be dispersed. The degree of dispersion varies with the wavelength. Then, for grating
type, dispersion is homogenous at the entire wavelength and a wide-range wavelength can be
obtained with a diffraction grating. In addition, a constant spectrum featuring a constant slit
breadth can be obtained. Next, the other component is sample container. A container that
contains a sample is usually called "cell" and two types are available that are glass and quartz
cells. Since light in the ultraviolet range with a wavelength of 340 nm or less hardly passes
through a glass cell, it is used for measurement in the visible range of 340 nm or more. On
the other hand, although a quartz cell allows passage of light in the entire wavelength in the
ultraviolet and visible ranges, it is mainly used for the measurement in the ultraviolet range
due to its high price. The last component is a detector. A detector plays a role in changing the
light transmitted from a sample into an electric signal. The detector is typically a
photomultiplier tube, phototube, photodiode and photodiode array. The maximum
wavelength of potassium permanganate that we obtained in this experiment is 525.0 nm. The
concentration of unknown solution of potassium permanganate that we obtained is 12.34
mg/L. In this experiment, some precautionary steps has been taken to avoid error from
occurred that will affect the results. For example, all the apparatus have been washed
properly with distilled water to avoid the solution from being contaminated with other
substances. Then, when I want to measure solution by using pipette and volumetric flask, my
eyes is perpendicular to the calibration mark to avoid parallax error from occurred.
Conclusion
As a conclusion, the calibration curve of potassium permanganate has been plotted. The
maximum wavelength of potassium permanganate that we obtained for this experiment is
525.0 nm and the concentration of an unknown solution of potassium permanganate is 12.34
mg/L.
References
1. (2013). In N. A. Faridah Hanum Hj Badrun, Basic Instrumental analysis Laboratory
experiments for an Introductory Course in Instrumental Analysis.
2. Zailuddin Ariffin, F. N. (2013). Basic Instrumental Analysis Laboratory Experiments
for an Introductory Course in Instrumental Analysis.
3. Scanning spectrophotometer. (n.d.). Retrieved from Chempage Laboratory resources:
http://chem.wisc.edu/deptfiles/genchem/lab/labdocs/modules/scanspec/scanspecdesc.
htm
4. UV-Vis/NIR Spectrophotometer Basic Course. (n.d.). Retrieved from Hitachi High-
Tech:
http://www.hitachi-hitec.com/global/science/uv_vis/uv_basic_8.html