Experiment 11: Quantitative Determination of Copper (II) Concentration by

Spectrophotometry
C. Grefaldia1, C. Hernia2
1 College of Home Economics, University of the Philippines, Diliman, Quezon City 1101
2 National Institute of Geological Sciences, College of Science, University of the Philippines, Diliman, Quezon City 1101

Performed 5 July 2017; Submitted 7 July 2017

Answers to Questions Moreover, limitation is due to the presence of stray

radiation. The radiation exiting from a monochromator is
1. What is the significance of the addition of ammonia to often contaminated with minute quantities of stray
radiation (radiation from the instrument outside the
Cu (II) solutions?
wavelength of band selected). This kind of radiation is
usually due to reflection and scattering by the surfaces of
This is performed to forward the reaction between copper
lenses, mirrors and windows. [3]
and ammonia to form a copper-ammonia blue complex in a
reaction as follows:
4. Why is it significant to scan over a wavelength range?
Why is the analytical wavelength used in the
Cu + 4NH3 [Cu(NH3)4]2+
determination of the absorbance of the standard and
sample solutions?
[Cu(NH3)4]2+ complex gives a more intense colour than a
standard copper solution which allows the
It is important to scan over a wavelength range in order to
spectrophotometer to examine it more effectively. [1]
maintain precision over measurement readings. Errors in
the wavelength scale of a spectrophotometer can have a
2. Why is Beer-Lambert Law expressed in terms of
significant impact on photometric measurements even
absorbance instead of transmittance?
when wavelength is not called out specifically as an
acceptance criterion for the analytical result.
The Beer-Lamberts law gives a convenient linear
relationship between the absorbed energy with the
5. Why do we have to measure absorbance reading
concentration of the absorbing species which makes the
against reagent blank solutions?
calculation easy.
This is to ensure that the readings are correct.
A = abc [2]
Spectrophotometer is set at a particular wavelength and
this wavelength is where the component being measured
where a is the absorptivity, b is the path length and c is the
absorbs the most. But, sometime the component being
concentration of the component.
measured contains other components beside the
component being measured. So, in order to minimize this
Since the absorptivity of the solution and the path length are
error in reading, a blank solution that doesnt contain the
kept constant throughout the experiment, we can establish
component being measured is measured and this provides
that absorbance is directly proportional to the
a base reading. Therefore, when the solution being
concentration of the analyte while transmittance is
measured is put in the instrument, the reading will be only
proportional to the intensity of the sunshine that has
due to the component. [1]
entered the pattern.
6. What is the significance of the y-intercept of your
A = log 1/T [1]
calibration curve? Discuss its deviation from the
theoretical value.
3. What are the limitations of the Beers Law?
Utilizing the equation y= mx + b, absorbance could be
controlled by comparing it to y. this is determined by the
One limitation to Beers Law is the assumption that the
spectrophotometer. X is equal to the concentration of the
radiation that reaches the sample is of a single wavelength
solution, m is equal to the absorptivity and b is the y-
that is, that the radiation is purely monochromatic. Beers
intercept. In instances where there is no y-int , it signifies
Law is not applicable to polychromatic light since it causes
that there is no deviation from the theoretical value. In the
negative deviation to Beers Law.
y-int, there is no concentration of Cu(II) in light of the fact
that x = zero. Hypothetically, the absorbance now would be
Another limitation is that at high concentrations, solute
zero on the grounds that there is no coloured solutuon
molecules can cause different charge distribution on their
because of the absence of copper which implies that no light
neighboring species in the solution. Since UV-visible
ought to be absorbed because of the solution being vapid or
absorption is an electronic phenomenon, high
colourless. The presence of a y-int even in the wake of
concentrations would possibly result in a shift in the
discrediting the affecting of the cell and the alkali solution
absorption wavelength of the analyte. At times, even
on the estimations of absorbance means that something else
electrolyte concentrations (such as those present in buffers)
is retaining light because of the presence of an absorbance
play an important role in altering the charge distributions
value even at zero ppm concentration. For this situation, a
and affecting UV-visible absorbance.
y-int would deviate from the theoretical value by expanding
the estimations of absorbance. [1]
 [4] Skoog et al, Applications of Absorption Spectroscopy.
7. Cite other analytical applications of Pharmatutor.org. n.d. 2010 Retrived 6 July 2017
spectrophotometry.

Spectrophotometry is applied in different fields. In

microbiology, it is used in order to quantify the number of
microbes in a specimen and detect impurities. For instance,
benzene, an impurity found in cyclohexane, can be
distinguished at 255nm. [1] The dissociation constants of
acids and bases can likewise be determined
spectrophotometrically from graphing absorbance and
wavelength at various pH values. Drugs can be
quantitatively analyzed by making a solution of drug and
measuring the absorbance at a particular wavelength.
Chemical kinetics can also be examined through
spectrophotometry. This is performed by measuring
absorbance changes based on UV radiation passing a
reaction cell. [4]

8. What are the possible sources if errors and their

effect on the calculated parameters? Rationalize.

Possible sources of errors for this experiment would be

random, systematic and gross. Random errors are often
unavoidable. They result from the use of glassware and may
influence the precision of the data. Systematic errors could
happen from method or instrument. The absorbance value
gotten from the spectrophotometer may increase if the
reaction cell was held improperly. This is a direct result of
the fingerprints might be left in the cell. These fingerprints
may absorb light from the spectrophotometer which builds
absorbance value. Since a similar cell was utilized for all
tests in this trial, it is essential that it is washed with the
solution going to be tested thoroughly. This is done to
guarantee that the right concentration of the Cu(II) is being
analyzed or the absorbance value may increase or diminish
relying upon the contaminated concentration. The reagent
blank test must be performed keeping in mind the end goal
to discredit the impacts of the cell and the ammonia solution
on absorbance or the absorbance value will increase.
Instrumental errors may occur in this experiment from the
spectrophotometer. Any stray light in the
spectrophotometer will decrease the absorbance value.
This is due to the additional presence of unabsorbed light.
The cell/cuvette used for analysis must be standardized
because a wider cell results in a longer path length. The
spectrophotometer should likewise be assessed before use
to guarantee it is as yet viable for examination. Gross
mistakes can come about because of contaminated
glassware particularly amid preparation of the solutions.
Undesirable reagents may enter the prepared stock
solutions and affect the concentration of copper which will
influence the absorbance value. This will influence the
development of the calibration curve which will affect the
estimations for the concentration of the unknown sample.
All concentrations must be uncontaminated before testing
to keep such mistakes from happening.

References
[1] Skoog,, Fundamentals of Analytical Chemistry, 8th ed.;
Brooks/Cole-Thomson Learning: California, 2004. Print
[2] UPD Chem 26.1 Analytical Chemistry Laboratory
Manual, Institute of Chemistry, University of the
Philippines, 2017 edition. Print.
[3] Tissue, Brian, Beer-Lambert Law, 2000
http://www.tissuegroup.chem.vt.edu/chem-
ed/spec/beerslaw.html Retrieved 6 July 2017