CHM 260 LABORATORY REPORT

Experiment 2: UV visible Determination of an Unknown

concentration of KMnO4 Solution

Name :
Student no :
Lab Group :
Date of report Submission :
Lab Partner’s Name :
Lecturer’s Name :
OBJECTIVES

1. To investigate the maximum wavelength of potassium permanganate

2. To plot the calibration curve of potassium permanganate
3. To determine the concentration of an unknown solution of potassium permanganate

INTRODUCTION

The spectrophotometer consists of light source, a sample compartment and a detector.

Monochromator is a device that used to separate light into its component wavelength.
Spectrophotometer can be measure into two ways which is scanning and diode array. The
diode array imparts light of all wavelengths to the sample at one time. The monochromator
consist of holographic grating that allow lights of individual wavelength to be sequentially
imparted to the sample.

Furthermore, when a beam of white light passed through a prism or reflected off a
diffraction grating, it is separated into a variety of colour, the rainbow phenomenon occurs
when the sunlight passes through the water droplet on a rainy day. The rainbow show variety
of colour can be described in term of their wavelength. The wavelength is a function of
energy of photon. Eyes detect the light and brain interprets the different wavelength and
energy. Short wavelength is at region in 350nm and is in the high energy. The long
wavelength was in 750nm that indicates the colour of red. For example, the long wavelength
such as infrared and radio waves and short wavelength is X-ray and the UV visible light.

Furthermore, the method of spectrophotometer is to use absorbance of light by an

analyte, potassium permanganate (KMnO4) at a certain wavelength to determine the
unknown concentration, the instruments is UV-Vis Spectrophotometer use UV light and
visible port of electromagnetic spectrum. It is able to effects the excitation of electron in the
atomic or molecular ground states energy to the higher energy levels which is resulting an
absorbance at specific wavelength to each molecule. The human eyes can see light in the
wavelength of 400nm to 700nm.

The correlation between wavelength, colour and complementary colour

 As the colour of KMnO4 purple, it absorbs the light in the green region of the
spectrum. Thus, human eyes see the transmitted light as a mixture of a violet and red that we
experience as a deep purple spectrophotometer in the visible region sources is a common
tungsten light bulb emitting white light that collimated and focused on slit entrance and then
fall on a chromator that separated white light in its constituent’s wavelength. From the
monochromator, the light is transfers through sample then finally reach at photocell. The
photocell measures the intensity of light at each specific wavelength and record the intensity
at that wavelength, I0(𝜆). The transmittance then display in the spectrophotometer.

𝐼
T=𝐼
𝑜

In the percentage, the transmittance defines as;

𝐼
T (%)=𝐼 × 100%
𝑜

For the diode array, the photocell measure light intensity of one wavelength at a time.
The spectrophotometer records the wavelength range between 200nm to 700nm within one
second. The Beer’s Law equation state that;

𝐼
A = -log 𝐼
𝑜

= -log (T)

= 𝜀𝑏𝑐

The 𝜀 is the molar absorptivity of the compound. Typically, it has unit of L mol -1 cm-
1
. The b is the path length in a unit of cm. the concentration of a compound define as c in the
unit of molarity mol/L. if the concentration has a unit of parts per million (ppm) then the 𝜀 is
ppm-1 cm-1. A is an absorbance symbols that directly proportional to the analyte
concentration.

The calibration curve technique consist of the known concentration of analyte that
place in the spectrophotometers and is recorded the data obtain. This response is corrected by
spectrophotometer output obtain with a blank solution. Then, the data is plotted by using
graph to determine the concentration of unknown. Analyte solution absorbs light at different
wavelength. For an instance, KMnO4 absorb light in a green region of the visible spectrum
and let the red and blue light passes through. Blue and red mixture is perceived by the eye as
the typical purple colour of KMnO4. Light which absorb arrange of wavelength is shown by a
spectrum. In a UV-Vis Spectrophotometer, we plot the graph of absorbance against the
wavelength reaching the solution. This is known as absorbance spectrum.
 In addition, analyte absorbance increases as the attraction of beam increases; it is show
from the Beer’s Lambert Law equations. When the molecules absorb the radiation at specific
wavelength, the absorption spectrum shows the number of absorption band that
corresponding to the structural group within the molecule. The absorption of UV visible
radiation corresponds to the excitation of an outer electron, electronic transitions have three
types that is;

i. Transition involving the 𝜋, 𝜎 and n electron

ii. Transition involving charge transfer electron
iii. Transition involving d and f electron

Electron undergoes excited states from ground states when atom or molecules absorbs
energy. Atom rotates and vibrates with respond to each other’s in molecules. The rotations
and vibrations has discrete energy levels which packed on top of each electronic level.

The chromophores consist of valance electron of low excitation energy. The molecule
spectrum containing the chromophores is complex due to the superposition of rotational and
vibrational transition and electronic transition give the overlapping lines combinations. The
different energy level as shown below;
Most UV-Vis spectrum involves 𝑛 → 𝜋 and 𝜋 → 𝜋 ∗ transitions are generally more intense
than𝑛 → 𝜋 ∗ . This is due to the absorption peaks for these transition falls in experimentally
convenient region of the spectrum (200nm-700nm). Hence, to provided 𝜋 electron, the
transition need to be unsaturated group in a molecule. The peaks 𝑛 → 𝜋 ∗ shifted to shorter
wavelength with increase solvent polarity that increased solvation of the lone pair which
lower the energy of n orbital. This is because the attractive polarisation force between solvent
and absorber lower the energy level of both excited and unexcited. The 𝑛 → 𝜎 ∗ transition
involving the saturated compounds consists of atom with the lone pairs. Usually, it need less
energy than𝜎 → 𝜎 ∗ . The number of organic functional group with 𝑛 → 𝜎 ∗ peaks in the
region is small. The energy requiring a large energy in the transition of 𝜎 → 𝜎 ∗ due to the
electron in bonding 𝜎 orbital is excited to corresponding anti-bonding orbitals.

APPARATUS

 Beaker
 Burette
 Glass rod
 Volumetric flask 100mL
 Dropper

CHEMICALS

 Potassium permanganate (KMnO4)

 Distilled water
PROCEDURE

A. Preparation of the KMnO4 Standard Solutions.

1. The solid of KMnO4 was weighted about 0.01g or 10mg using the weighing boat.
Then, the KMnO4 was transfer into the 100mL of volumetric flask using the filter
funnels.
2. A little bit of distilled water was pour into the volumetric flask to dissolve the
solid. Then, place the stopper on volumetric flask and was shake vigorously. The
distilled water once again pours until the calibration mark and shake the solution.
3. Then. The stock solution was pour into the beaker and was labelled as 100ppm
stock solution.
4. 5mL of stock solution was transferred into the 100mL of volumetric flask by
using burette. The distilled water was pour into the flask util the calibration mark.
5. The flask was label as 5ppm solution.
6. Step 4 and step 5 was repeated by using 10mL, 15mL and 20mL stock solution
and transfer into small beaker.
7. The beaker was labelled as 10ppm, 15ppm and 20ppm.

B. Preparation of the unknown.

1. The stock solution was pipet between 5 to 20 mL by using burette and transfer
into the flask. The distilled water was added until the calibration mark.
2. The stopper was placed on the flask and was shake until the solution was
homogenous.
3. The solution was transfer into the beaker and label as Unknown.

C. Determination of Absorption Maximum

1. The cuvette was obtain and clean and rinsed with distilled water and then was
filled with distilled water about ¾ of the cuvette.
2. Wipe the cuvette with Kimwipe
3. The cuvette was label as blank.
4. Step 1 and step 2 was repeated by using the 5ppm, 10ppm, 15ppm and 20ppm
with different cuvette
5. Label the cuvette as 5ppm, 10ppm 15ppm and 20ppm solution.

D. Operation of the UV-Vis Spectrophotometer

Instruments: Varian/ Cary 50 UV-Vis Spectrophotometer

Operation Instruction
1. The Cary win UV icon was select and click Scan
2. Click the setup, and start was set to 800, stop was 400 scan controls was fast, y
mode and y-min was set to absorbance and zero respectively.
3. The baseline was click and set to correction. Then, click report and filled in our
experiment title.
4. In the auto store icon, it was set to prompt at start. Then click ok.
5. The blank was place into the instrument and baseline was click then click ok.
After the baseline show it was zero click finish, print and then places the cuvette
with different concentration.
6. Then print the results.
7. The start icon was click and wavelength was set same as step 2. The replicates
were three.
8. In the standard icon, filled the standard and concentration. The fit type was linear
direct and filled in the number of sample.
9. Step 5 and 6 was repeated.

Data

Table of concentration and absorbance

Solution Concentration(ppm) Absorbance

Standard 1 5ppm 0.0796
Standard 2 10ppm 0.1546
Standard 3 15ppm 0.2374
Standard 4 20ppm 0.3133
Unknown ? 0.2108

Graph
 Concentration Against Absorbance
0.35

y = 0.0157x + 0.0003
0.3

0.25
Absorbance

0.2

0.15

0.1

0.05

0
0 5 10 15 20 25
Concentration(ppm)

Results, Calculation and Questions

Calculations

The sample preparation for the solution can be calculated by using the following formula M 1V1 =
M2V2.

M1V1 = M2V2

Standard 1, (100ppm)(V1) = (5ppm)(100mL)

(5ppm)(100mL)
(V1) = (100ppm)
 = 5mL

Standard 2, (100ppm)(V1) = (10ppm)(100mL)

(10ppm)(100mL)
(V1) = (100ppm)

= 10mL

Standard 3, (100ppm)(V1) = (15ppm)(100mL)

(15ppm)(100mL)
(V1) = (100ppm)

= 15mL

Standard 4, (100ppm)(V1) = (20ppm)(100mL)

(20ppm)(100mL)
(V1) = (100ppm)

= 20mL

Based on the graph the slope of the graph of absorbance against concentration of potassium
permanganate was m = 0.0157. the molar absorbtivity was equal to the slope of graph. The
path length was 1 cm and the absorbance was 0.2108. according the Beer’s Law, the formula
to find concentration of the unknown was;

A = Ɛbc
𝐴
C = Ɛb

0.2108
C = (0.0157𝑝𝑝𝑚−1 𝑐𝑚−1 )(1𝑐𝑚)

= 13.4ppm

Questions

1. Why is glass in not suitable cell material for use in UV spectroscopy?

- Glass will absorb some of the ultra violet light and will cause an inaccurate result for
the experiment as the reading will be higher than the exact one.
2. State one advantage of using the UV-Vis Spectrophotometer compared to a
Spectronic 20 for this analysis
- The advantage is to record absorbance at each wavelength and rapidly scan a range of
wavelength
Discussion

In these experiments, the technique was done to determine the maximum wavelength
of potassium permanganate of stock solution. The maximum wavelength recorded was plot
the calibration curve by using the UV-Vis instruments. Hence, we can also determine the
unknown concentration of potassium permanganate for this experiment. First, the solid of
potassium permanganate was weighted roughly about 10mg on weighing boat by using
electric balance. The 10mg potassium permanganate was dissolve in the volumetric flask by
added small quantities of distilled water. The flask was seal with stopper and shakes the flask
vigorously. It was important to shake the flask because to make the solution homogenous.
Then, distilled water was pour in the flask until the calibration mark and was shake once
again. The 100ppm of potassium permanganate was label as stock solution. By using the
stock solution, 5mL, 10mL, 15mL and 20mL was prepared by pipet 5mL, 10mL, 15mL and
20mL into the different flask and add distilled water until the calibration mark. The volume
of stock solution needed was calculated by using the formula M1V1 = M2V2. The solution was
transfer into the sample bottle and label was 5ppm, 10ppm, 15ppm and 20ppm respectively.
The unknown was prepared with the same method as standard solution but the amount to
pipet from stock solution must be between 5mL to 20mL. Then, the unknown solution was
label as unknown.

For the next part of the experiment, we use the UV-Vis instrument to determine the
maximum wavelength of potassium permanganate. The cuvette was filled with distilled water
about ¾ of cuvette volume. Then, the other standard was prepared by filled the cuvette with
different amount of concentration of potassium permanganate. The instrument was setting
based on the procedure. After finish setting up, the blank solution was place into the
instrument and the baseline was click. After the blank solution became zero, the standard
solution of 5ppm was placed in UV-Vis Spectrophotometer and the maximum wavelength
was determined. The maximum wavelength is where the wavelength with higher absorbance.
The data shows that, the maximum wavelength obtained was 525nm at absorbance of 0.316.

The graph that we plot concentration against absorbance resulting the linear
exponential line. Each point was representing the wavelength and absorbance of standard1, 2,
3, 4 and unknown respectively. For the 5ppm concentration, the absorbance that we obtained
was 0.0796. The absorbance of 0.1546 was the concentration of 10ppm. The 15ppm indicates
the absorbance was 0.2374. For 20ppm, the absorbance was 0.3133 and for the unknown
concentration obtains the absorbance of 0.2108. The slope that we obtain was 0.0157 which
indicates the molar absorptivity of the potassium permanganate. By using the Beer’s Law
formula of A=𝜀𝑏𝑐, we can calculated the concentration of unknown. The b represent the path
length which is 1cm. thus, the concentration of unknown was 13.4 ppm by using this formula.
Based on the spectrogram conducted, the concentration of unknown was same, 13.4ppm as
calculated.
 Furthermore, the graph showed the R2 value that is the correlation coefficient value.
Correlation coefficient value is the statistical measure of the degree to which changes to
the value of one variable predict change to the value of another. The correlation coefficient
varies about +1 to -1. The graph showed the correlation efficient of 0.95000 and the
spectrogram resulting the R2 of 0.99982. The value of 0.99982 is the closest to +1 which
indicates the accurate results that we got. Although the graph correlation coefficient was
0.95000, but it still closest to the theoretically value as mentioned.

Conclusion

According to these experiments, the absorbance of potassium permanganate was 0.0157 ppm-
1
cm-1 and the maximum wavelength was 525nm. The Beer’s Law equation used to determine
the concentration of unknown solution that was 13.4ppm with the correlation efficient of
graph and spectrogram was 0.95000 and 0.99982 respectively.

References

David B. Green(1996). Quantitative Chemistry and Instrumental Analysis. Retrieved date by

15 July 2014.

L.E. Laverman. (n.d).Experiments in Analytical, Physical and Inorganic Chemistry – 3rd

Edition. Retrieved date by 15 July 2014.

Robert Bohman. (2006). Ultraviolet/Visible (UV-Vis) Spectroscopy of Potassium

Permanganate. Retrieved date by 15 July 2014.