CHM260

BASIC INSTRUMENTAL ANALYSIS

LABORATORY SUMMARY WRITTEN

REPORT

Name : NURUL SHUHADAH BINTI

ABDUL HADI
ID No. : 2018637498
Programme : AS120 (5J)
Instructor : MADAM ROSLIZA BINTI ALI
 EXPERIMENT 2
UV-Visible Determination of an unknown concentration of KmnO4 Solution
A. Pre-laboratory questions

a) Explain how you prepare a 5ppm solution from a 100ppm stock solution using a 100
mL volumetric flask.
5 mL of the ’stock’ solution has been diluted with distilled water into the 100 mL
volumetric flask until the calibration mark. Then, the solution has been transferred into
a beaker.
b) What is the expected wavelength at maximum absorption (max) for KMnO4? What is
the absorptivity () value of this wavelength? (please provide the reference used to
answer the question)
The maximum absorption peaks for permanganate appeared at 310 and 530 nm
(Hamada, 2016). Molar absorptivity values are calculated to be 2.33∙104 and
2.66∙104 l/mol cm (Devi, 2012).

B. Post-laboratory questions

a) Why are glass materials not suitable for UV-spectroscopy cell holder?
Glass absorbs strongly in UV region and its application is not recommended for
wavelengths below 340 nm.
b) State one advantage of using the UV-Vis spectrophotometer compared with Spectronic
20 for this analysis.
To record absorbance at each wavelength and rapidly scan a range of wavelength.

C. Complete the table of concentration and absorbance

Solution Concentration Absorbance
Standard 1 5 0.135
× 100 = 5 𝑝𝑝𝑚
100
Standard 2 10 0.278
× 100 = 10 𝑝𝑝𝑚
100
Standard 3 15 0.418
× 100 = 15 𝑝𝑝𝑚
100
Standard 4 20 0.562
× 100 = 20 𝑝𝑝𝑚
100
Unknown A = εbc 0.251
0.251 = 0.0284 (1.00)(C)
C = 8.84 ppm
D. Report summary (1-2 pages)
a) Brief Introduction of the experiment
Ultraviolet and visible light range (UV/VIS) is widely applied in research,
production and quality control for the classification and study of substances.
UV/VIS spectroscopy is based on the absorption of light by a sample. Depending
on the amount of light and its wavelength absorbed by the sample, valuable
information can be obtained, such as the purity of the sample. Moreover, the
amount of absorbed light is related to the amount of sample, and thus, quantitative
analysis is possible by optical spectroscopy. A UV/VIS spectrophotometer
measures the intensity of light passing through a sample solution in a cuvette, and
compares it to the intensity of the light before it passes through the sample. The
main components of a UV/VIS spectrophotometer are a light source, a sample
holder, a dispersive device to separate the different wavelengths of the light and a
suitable detector. This instrument measures transmittance and the absorbance
which is defined as A = −log(Transmittance). In general, a UV/VIS spectrum is
graphically represented as absorbance as a function of wavelength. The advantage
of this representation is obvious; the height of the absorption peaks is directly
proportional to the concentration of the species. The calculation of concentration is
governed by the Lambert-Beer Law. To calculate the concentration: C = A / ε
(epsilon) x b where C = The sample concentration in mol / L or g / mL, b = Cuvette
path length in cm ε = (epsilon) sample specific constant (describing how much the
sample absorbs at a given wavelength). The ultraviolet region (about 400-190 nm)
is particularly important for the qualitative and quantitative determination of many
organic compounds, especially those with a high degree of conjugation, also absorb
light in the UV or visible regions of the electromagnetic spectrum. The solvents for
these determination are often water for water-soluble compounds, or ethanol for
organic-soluble compounds. While in the visible region (about 400-820 nm).
Spectrophotometry methods are widely used for the quantitative determination of
many trace substances, especially inorganic species.
b) State the experiment methodology
A. Preparation of the KMnO4 Standard Solutions
1. 0.01 g of KMnO4 has been weighed accurately to the nearest mg, on a weighing
paper. The reading has been recorded. Using a funnel, the solid has been
transferred to a 100 mL volumetric flask.
2. The solid has been dissolved with a few mL of distilled water. The flask has
been stoppered and shook. Distilled water has been added to the mark, using a
medicine dropper to add the last few drops. The flask has been stoppered and
shook several times to homogenize the solution.
 3. The ‘stock’ solution has been poured into a beaker. The beaker has been labelled
as ‘100 pm’.
4. 5.00 mL of the ‘stock’ solution has been pipetted and diluted with distilled water
in a 100 mL volumetric flask.
5. The solution has been transferred into a beaker and label as ‘5 ppm’.
6. Step 4 has been repeated, using 10 mL, 15 mL and 20 mL stock solution and
transferred into small beakers.
7. The beakers have been labelled as ’10 ppm’, ’15 ppm’ and ’20 ppm’
respectively.
B. Preparation of the Unknown
1. Between 5.00 to 20.00 mL of the ‘stock’ KMnO4 solution has been pipetted and
diluted with distilled water in a 100 mL volumetric flask.
2. The solution has been transferred into a beaker and labelled as ‘Unknown’.
C. Determination of Absorption Maximum (max)
1. Both cuvettes were filled with blank solution (distilled water).
2. The clear side of the cuvette has been placed facing the correct light source
direction.
3. The lid has been closed.
4. 0% transmittance and absorbance have been adjusted.
5. One of the cuvette has been filled with 20 ppm standard solution.
6. It has been placed in the sample compartment.
7. The maximum wavelength has been scanned.
D. Determination of Standard Solution and Unknown Sample’s Absorbance
1. One of the cuvette has been filled with 5 ppm standard solution.
2. It has been placed in the sample compartment to scan for the absorbance.
3. The steps for all standard and unknown sample have been repeated.
c) State your finding (absorbance spectrum/calculation) and briefly discuss the
finding.
Solution 1
5
× 100 = 5 𝑝𝑝𝑚
100
Solution 2
10
× 100 = 10 𝑝𝑝𝑚
100
Solution 3
15
× 100 = 15 𝑝𝑝𝑚
100
Solution 4
20
× 100 = 20 𝑝𝑝𝑚
100
Unknown solution
A = εbc
0.251 = 0.0284 (1.00)(C)
C = 8.84 ppm
The unknown concentration of KMnO4 solution can be determined using UV-Visible
Spectrophotometer. The standard solutions of KMnO4 can be prepared using 5 mL, 10
mL, 15 mL and 20 mL of the ‘stock’ solutions. The concentration in ppm can be
𝑣𝑜𝑙𝑢𝑚𝑒 (𝑖𝑛 𝑚𝐿)
calculated using formula, × 100 𝑝𝑝𝑚. The calculated concentration for
100 𝑚𝐿
the 0.135 absorbance was 5 ppm. The calculated concentration for 0.278 absorbance
was 10 ppm. The calculated concentration for the 0.418 absorbance was 15 ppm. The
calculated concentration for the 0.562 absorbance was 20 ppm. The plotted graph of
concentration (ppm) versus absorbance resulting in a liner exponential line, which
 means the concentration is directly proportional to the absorbance. The molar
𝑦2−𝑦1
absorptivity, ε can be obtained by using the slope of the graph, 𝑚 = resulting in
𝑥2−𝑥1
the value of 0.0284. Thus the unknown concentration of the KMnO4 solution can be
calculated by filling in the value into the formula of Beer’s Law, A=εbc and the
resulting value for the concentration was 8.84 ppm for the 0.251 absorbance. The
maximum wavelength, max of the KMnO4 solution was 525.6 nm. The plotted graph
showed the R2 value = 1 that was the correlation coefficient value. Some precaution
steps must be taken while performing this experiment - when using
spectrophotometers, all of the data must be taken on the same instruments, at the same
time, if changing instrument was needed, then data for the spectrum must be taken and
start over again to avoid inaccurate values.
d) State the advantages and limitation (if any) of the instrument used to conclude the
experiment conducted
The biggest advantage of the UV-VIS spectrometers is the accuracy of the device.
Even small UV-VIS spectrometers can give extremely accurate readings, which is
crucial when you are preparing chemical solutions or recording the movement of
celestial bodies. UV-VIS spectrometers are easy to use. Most of the ones used in
chemistry are comparable in size to electron microscopes and require the same basic
skills to use. Because they are simple to operate, there is little chance of a UV-VIS
spectrometer being used improperly. The main disadvantage of using a UV-VIS
spectrometer is the time it takes to prepare to use one. With UV-VIS spectrometers,
setup is key. Area of any outside light, electronic noise, or other outside contaminants
must be cleared as that could interfere with the spectrometer's reading. If the space
has not been properly prepared, even a small bit of outside light or vibration from a
small electronic device could interfere with the results.
e) Cited 2-3 references used.

References
Cook, M. (2018, April 29). Sciencing. Retrieved from https://sciencing.com/advantages-disadvantages-
uvvis-spectrometer-6466475.html

Devi, O. Z. (2012, January 17). SpringerLink. Retrieved from

https://link.springer.com/article/10.1007/s10812-012-9547-9

Hamada, Y. Z. (2016, November 16). eJBio. Retrieved from https://ejbio.imedpub.com/three-very-

different-uvvis-absorption-spectra-of-threedifferent-transition-metals-found-in-biological-
solutions.pdf