CHM260
BASIC INSTRUMENTAL ANALYSIS
LABORATORY SUMMARY WRITTEN
REPORT
Name : ILYA ZAFIRAH BINTI ALI
ID No. : 2018699856
Programme : AS120
Instructor : SITI NORHAYATI BT. MOHD
TARMIZI
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 can be defined as a study of the interaction between electromagnetic (EM)
radiation and matter. The word spectroscopy is used to refer to the broad area of science
dealing with absorption, emission or scattering of electromagnetic radiation by molecules,
ions, atoms or nuclei. The type of electromagnetic radiation used in this experiment is visible
light.
b) Define the terms of transmittance and absorbance.
Transmittance can be defined as the fraction of incident radiation transmitted through
the sample medium. Absorbance is 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 𝜆𝑚𝑎𝑥 .
Beer’s Law can be equated with 𝐴 = 𝜀𝑏𝑐 where A is absorbance, ɛ is molar
absorptivity with units of L mol-1 cm-1, b is the path length of the sample in cm, and c
is concentration in mol L−1 . 𝜆𝑚𝑎𝑥 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
Increase
ii. Transmittance
Decrease
b) Why essential to obtain the absorption spectrum of the soft drinks first before
developing a calibration curve?
Obtaining the absorption spectrum before making the calibration curve is essential
because the wavelength of maximum absorbance for a given sample can be selected
and used in absorbance measurements.
c) What is the purpose of using the ‘blank’ solution?
The 'blank' allows you to set the spectrophotometer to zero before you measure your
'unknown' solution. The 'blank' solution will contain everything that the
'unknown' solution (the one you want to measure) except for the think you wish to
measure.
d) Name the colours absorbed in soft drink samples.
Blue-green
C. Complete the following table with wavelength and absorption given by the
instructor
Wavelength (nm) Absorbance
600 0.040
580 0.214
560 0.574
540 0.770
520 0.885
500 0.796
480 0.610
460 0.383
440 0.252
420 0.213
400 0.207
380 0.194
360 0.200
D. Complete the table of soft drinks concentration (volume %) and absorbance
Solutions Concentration (%) Absorbance
1 10 0.179
2 20 0.362
3 30 0.537
4 40 0.714
5 50 0.885
Unknown sample 14 0.245
E. Report summary (1-2 pages)
a) Brief Introduction of the experiment
The word spectroscopy is used to refer to the broad area of science dealing with the
absorption, emission, or scattering of electromagnetic radiation by molecules, ions,
atoms, or nuclei. Spectroscopic techniques are some of the most widely used analytical
methods in the world today. Different regions of the electromagnetic spectrum such as
infrared, visible, ultraviolet, or X-ray radiation can be used to interact with matter.
There are many important instrumental methods of chemical analysis that rely on the
interaction between light and matter to probe chemical structure.
White light is visible light that contains all the colours of the rainbow. If white light hits
a coloured solution, the electrons in the solution will absorb some of that, while the rest
will be transmitted through the solution. It is the transmitted light that we see with our
eyes. It is important to understand that a red solution does not absorb red light; it
actually absorbs blue-green light. The solution appears red because the intensity of the
blue-green light in the original white light has been decreased. The following table
shows the relationship between the colour absorbed by a solution, and the colour we
observe.
b) State the experiment methodology
To prepare the standard solution, soft drink was poured into a beaker and it was stirred
to remove carbonation. 5.00mL of the soft drink was pipetted into a 50.00mL
volumetric flask and it was diluted with distilled water up to the mark. The solution was
shaken to homogenize it. This step was repeated using 10.00mL, 15.00mL, 20.00mL
and 25.00mL of the soft drink. To prepare the unknown solution, the soft drink was
poured without measuring the volume into a 50.00mL volumetric flask and it was
diluted with distilled water up to the mark.
Next, to determine the maximum wavelength, a cuvette was obtained, cleaned and
rinsed with distilled water. It was filled about ¾ full of blank solution (distilled water).
Any solution or fingerprints were carefully wiped away with a Kimwipe. The cuvette
was placed in the sample compartment and the clear side of the cuvette was made sure
to face the direction of the light source. 0% absorbance and 100% transmittance was
adjusted. Another cuvette was cleaned and rinsed with a small amount of the standard
solution. Any of the standard solution made previously was used. The standard solution
was filled into the cuvette about ¾ full and it was wiped with a Kimwipe. The cuvette
was placed in the sample holder and the absorbance was recorded. The cuvette was
removed. By referring to the lab manual, the wavelength was changed. The cuvette with
distilled water was inserted in the sample compartment and the absorbance was reset to
0% while the transmittance to 100%. The cuvette was removed and the cuvette
containing the same soft drink standard solution used previously was inserted. The
absorbance was recorded and this step was repeated with all wavelength.
Then, to determine the absorbance of the standard solutions and unknown sample, the
spectrophotometer was set to the maximum wavelength obtained from the previous
part. A cuvette was cleaned and rinsed with the most diluted standard solution and it
was filled with it up to ¾ full. It was cleaned with a Kimwipe. The cuvette with distilled
water was inserted into the sample compartment and reset 0% absorbance and 100%
transmittance. The cuvette was removed and the reading was recorded. The step was
repeated for all of the standard and unknown solutions.
c) State your finding (figures/calibration curve/calculation) and briefly discuss the finding
Absorbance vs. Wavelength
0.9
0.8
0.7
0.6
Absorbance
0.5
0.4
0.3
0.2
0.1
0
350 400 450 500 550 600 650
Wavelength (nm)
The maximum absorption for the soft drink is at 520nm with absorbance of 0.885.
The 𝜆𝑚𝑎𝑥 is 520nm. The colour of the soft drink is red, hence the absorbed colour is
blue-green.
Beer's Law Plot
1
0.9
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
0 10 20 30 40 50 60
y2 −y1 0.362−0.179
m= = = 0.0183
x2 −x1 20−10
y = mx + c
0.179 = (0.0183)(10) + c
0.179 = 0.183 + c
c = -0.004
The concentration of unknown sample:
y = mx + c
0.245 = (0.0183)x + (-0.004)
0.249 = 0.0183x
x = 13.6%
The concentration of the unknown sample is 13.6%
d) State the advantages and limitation (if any) of the instrument used as a conclusion.
The advantages of using Spectronic 20 are it is suited for quantitative absorption
measurements at a single wavelength and it is easy to maintain. The limitations of this
device are two separate readings has to be made on the light and changing of wavelength
is accompanied by a chance in light intensity.
e) Cited 2-3 references used.
i. https://www.acttr.com/en/en-faq/en-faq-uv-vis/134-en-faq-uv-vis-advantage-
disadvantage.html
ii. https://sciencing.com/difference-between-spectrometer-spectrophotometer-
8577067.html
iii. https://askinglot.com/what-is-the-purpose-of-the-blank-solution-in-a-beers-
law-experiment
