Experiment 1 CHM260 PDF

EXPERIMENT 1:
The Visible Spectra of Soft Drinks

(Refer to Lab manual Exp. 1 and lecture notes to answer the following pre and post laboratory
questions)
A. Pre-laboratory questions.
a) Define spectroscopy and state type of electromagnetic radiation used in this experiment.
Spectroscopy is the study of the relationship and interaction between electromagnetic radiation and
matter. The type of electromagnetic radiation used was visible light.
b) Define the terms of transmittance and absorbance.

The terms of transmittance refer to the fraction of incident radiation transmitted through the sample
medium while absorbance refers to a measurement of the amount of radiant power absorbed by the
sample defined as the -ve log of T.
c) State Beer’s law mathematically and max.

Beer’s law mathematically is A = εlc and max refers to the wavelength along the absorption
spectrum where a substance has its strongest photon absorption.
B. Post-laboratory questions
a) Based on Beer’s Law, when the concentration of an analyte increases, how will the
following be affected (increase, decrease, no change).
i. Absorbance (As A = abc so, it will increase)
ii. Transmittance (As A = -log T so, it will decrease)
b) Why essential to obtain the absorption spectrum of the soft drinks first before
developing a calibration curve?
It is to select the wavelength of maximum absorbance and use it in the absorbance measurements.
c) What is the purpose of using the ‘blank’ solution?

The ‘blank’ solution is a solution that contained a very little amount analyte of interest used to
calibrate instruments and set the wavelength to 0 nm before we measure the ‘unknown’ solution.
d) Name the colours absorbed in soft drink samples.

Based on the lambda max, the colour absorbed in soft drink samples was blue-green.
C. Complete the following table with wavelength and absorption given by the instructor.
Wavelength (nm) Absorbance
600 0.029
580 0.077
560 0.202
540 0.286
520 0.323
500 0.300
480 0.239
460 0.166
440 0.149
420 0.137
400 0.142
380 0.162
360 Too dark
D. Complete the table of soft drinks concentration (volume %) and absorbance.

Solutions Concentration Absorbance
1 10 0.340
2 20 0.618
3 30 0.929
4 40 1.230
5 50 1.560
Unknown sample 38 1.197
E. Report summary (1-2 pages)

a) Brief Introduction of the experiment.
Spectroscopy is the study of the relationship and interaction between electromagnetic radiation and
matter. This includes infrared (IR), ultraviolet (UV), and X-Ray radiation. Spectronic 20 which
used visible light spectra was used in this experiment.
b) State the experiment methodology.
A. Preparation of ‘Standard’ Solutions of Soft Drink (Known Concentrations).
1. Soft drink was poured into a beaker and the carbonation was removed.
2. 5.00 mL of the soft drink was pipetted into a 50.0 mL volumetric flask and was diluted to
the mark with distilled water. It was then covered and shaken to make a homogenous
solution and was stored in a small beaker.
3. Step 2 was repeated using 10 mL, 15 ml, 20 mL, and 25 mL of soft drink.
B. Operation of The Spectronic 20 and Determination of max.
1. The Spectronic 20 was turned on and warm-up for a minimum of 15 minutes.
2. The wavelength was set to 600 nm and 0% transmittance was adjusted and a cuvette was
obtained. The tube was cleaned and rinsed with distilled water and was filled with ¾ full
of ‘blank’ solution.
3. 0 absorbance and 100% transmittance were adjusted with the cuvette that contained the
‘blank’ in the sample holder. The cuvette was then removed and it was set aside without
emptying the distilled water.
4. Another cuvette was cleaned and rinsed with a small amount of ‘standard’ soft drink that
its absorbance will be observed. Then, it was filled of ¾ full with solution. Next, it was
wiped with Kimwipe and placed in the sample holder with harsh mark aligned.
5. The absorbance recorded. The cuvette was removed, the top was closed and the wavelength
changed to a setting which 20 nm lower. 0% transmittance was reset if it had changed.
6. The cuvette of distilled water was inserted and 100% T was reset, then the cuvette was
removed. The cuvette that contained the same Soft Drink used in Step 6 was inserted. The
absorbance was read and recorded in Table 1.2.
7. Steps 6 were repeated until 360 nm, absorbance readings were taken at each 20nm interval.
The absorption spectrum of the soft drink was plotted and the max was determined by using
the graph paper.
C. Preparations of ‘Unknown’ Soft Drink Sample.
1. The soft drink was poured into a beaker and the carbonation was removed by stirring.
2. The soft drink was poured without measuring the volume into a 50 mL volumetric flask
and was diluted to the mark with distilled water.
3. Stopper was used and the flask was shaken to homogenize the solution. The colour of the
unknown solution was made sure, not darker than the most concentrated standard solution.
4. The ‘unknown sample’ was put into a cuvette until ¾ full.
D. Quantitative Analysis of The Soft Drink Solution.
1. The Spectronic 20 was set to the wavelength maximum obtained from Part B.
2. 0 and 100%T was set and the absorbance of each 5 ‘standard’ soft drink solution was
measured and recorded while the absorbance of the ‘unknown’ soft drink solution was
measured and recorded.
E. Cleaning Up.
1. The waste was poured down the drain, and all cuvettes were cleaned and dried.
c) State your finding (figures/calibration curve/calculation) and briefly discuss the finding.
Graph Absorbance Vs Wavelength(nm)

0.35 0.323
0.3
0.286
0.3
0.239
0.25 0.202
Absorbance
0.2 0.166
0.149 0.137 0.142 0.162
0.15
0.1 0.077
0.029
0.05 0
0
max
600 580 560 540 520 500 480 460 440 420 400 380 360
Wavelength (nm)
Figure 1.1
Graph Absorbance Vs Concentration (volume%)
1.8
1.56
1.6
1.4 1.23
1.2 y = 0.0305x + 0.0198
Absorbance
0.929 R² = 0.9992
1
0.8 0.618 1.197 = 0.0305x + 0.0198
0.0305x = 1.197 - 0.0198
0.6
0.34 x = 38.60
0.4
0.2
0
0 10 20 30 40 50 60
Concentration (%)
Figure 1.2
Discussion.
Based on figure 1.1, the max was determined to be 520nm while the unknown concentration with
the absorbance of 1.197 is 38.6%, obtained in figure 1.2. The theory stated that visible light absorbs
blue-green light, so it was known that the practical experiment was conducted correctly.
d) State the advantages and limitations (if any) of the instrument used as a conclusion.
In conclusion, the advantages are its high throughput and high sensitivity and not to forget that it is
less complicated for students to conduct the experiment. However, this single beam type of
spectrophotometer is a significant amount of time needed for taking the reference and making
sample measurement hence it takes a long time to conduct the experiment. Moreover, the origin of
the colour absorbed in the soft drink is blue-green, the lambda max is 520nm while the unknown
concentration of a soft drink from the calibration curve is 38.60%.
e) Cited 2-3 references used.
1. Sheffield Hallam University, accessed on 3 November 2020,
https://teaching.shu.ac.uk/hwb/chemistry/tutorials/molspec/beers1.htm
2. LibreTexts libraries, MindTouch Copyright, accessed on 1 November 2020,
https://chem.libretexts.org/Bookshelves/Physical_and_Theoretical_Chemistry_Textbook_Ma
ps/Supplemental_Modules_(Physical_and_Theoretical_Chemistry)/Spectroscopy/Electronic_
Spectroscopy/Electronic_Spectroscopy_Basics/The_Beer-Lambert_Law
3. Khan Academy, accessed on 4 November 2020,
https://www.khanacademy.org/science/organic-chemistry/spectroscopy-jay/uv-vis-
spectroscopy/v/absorption-in-the-visible-region

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EXPERIMENT 1:

The Visible Spectra of Soft Drinks

b) Define the terms of transmittance and absorbance.

c) State Beer’s law mathematically and max.

c) What is the purpose of using the ‘blank’ solution?

d) Name the colours absorbed in soft drink samples.

D. Complete the table of soft drinks concentration (volume %) and absorbance.

E. Report summary (1-2 pages)

Graph Absorbance Vs Wavelength(nm)

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