DETERMINATION OF COPPER (II) CONCENTRATION BY

COLORIMETRIC METHOD

N. D. E. SILAVA

INSTITUTE OF BIOLOGY, COLLEGE OF SCIENCE

UNIVERSITY OF THE PHILIPPINES, DILIMAN, QUEZON CITY, PHILIPPINES

DATE PERFORMED: SEPTEMBER 22, 2010

ABSTRACT

The experiment was done to determine the Cu (II) concentration in a sample

by using a Spectrophotometer and by applying Beer’s law. A calibration curve was
obtained by plotting the absorbance against concentration of the standard Cu(II)
solution and the concentration of the unknown solution was determined using the
equation of the line of the calibration curve. Calculated concentration of unknown
sample yielded a 13% deviation which meant that colorimetry yielded acceptable
results. Results obtained were in agreement with Beer’s law stating that
absorbance is directly proportional to concentration.

INTRODUCTION called the Beer-Lambert law, the

Colorimetry is defined as the
method of trace analysis that involves
absorption of radiations having
frequencies in the visible spectrum.
[1] this is done using a
Spectrophotometer that transmits
radiation from a source through a
sample cell and into a detector which
measures the absorbance of the
sample. [1] The wavelength of
radiation absorbed depends on the
identity and amount of substance
present in the sample. [2] The
relationship between these factors is
Bouger-Beer law or more simply,
Beer’s law.[3] which is given by
-log(T)=-log(I/Io)=εbc=A
Where T is the transmittance of the
beam or the fraction of the incident
radian power transmitted by the
sample (I/Io), “ε” is the molar
absorptivity and is replaced by the
symbol “a” if concentration is given in
grams/liter. “b” is the path length , “c”
is the concentration of the sample and
“A” is the absorbance. [3] [4] [5]

1
In this experiment, a UV-Vis
spectrophotometer was used for the
analysis of the sample. First, solutions
of 0 ml, 2 ml, 4 ml, 6 ml 8 ml and 10
ml were taken from a 2500 ppm
working solution and placed in 6 50
ml-volumetric flasks. 10 ml ammonia
solution was added to each flask and
then diluted to mark. The blank
solution was put in a plastic cuvette
and into the spectrophotometer to
minimize the error caused by the
reflected light [3] then the analytical
wavelength was set by using the most
concentrated working standard. The
calibration curve was then prepared
by using the prepared solutions and
plotting their Absorbance (A) vs
Concentration (ppm Cu). Then, a
sample of unknown concentration was
obtained from the instructor and 3
absorbance readings were taken and
the concentration was determined.
Thus, it can be said that this
experiment was done to determine the
concentration of Cu (II) ions in a
sample by taking the absorbance and
using Beer’s law to compute the
concentration.
RESULTS AND DISCUSSION
The determined analytical
wavelength (λ) was 606nm for
optimum absorption. The blank
solution was then used to reset the
values and take into account the
transmitted radiation lost due to
scattering, reflection, etc.[3] Ammonia
was then added to the samples since
Cu is weakly colored to be measured
and thus, the ammonia reacts with the
copper to form a deep blue complex.
2
Cu2+→Cu(NH3)42+ (deep blue) [2]
The values of absorbance (A) for a
2500 ppm Cu (II) standard solution
were then measured.
Volume Std(ml) Cu (II) ppm Absorbance
2 100 0.082
4 200 0.17
6 300 0.268
8 400 0.366
10 500 0.456
Table 1. Absorbance of solution
It was Absorbance was then plotted
versus Concentration in parts per
million (ppm) and the equation of the
line was determined. It was necessary
to use absorbance rather than
transmittance since absorption peaks
appear as deep valleys in a
transmittance plot whereas an
absorbance plot would produce a
linear result. [3]
Figure 1. Calibration curve
Then concentration (x) was expressed
in terms of (y) giving a working
equation
x=y+0.0148÷0.009
This determined the concentration of
the unknown solution given a value for
absorbance (Y). It could be said that
the experimental points obey Beer’s
law since they lie on a straight line,
giving a slope that represents the
product of the molar absorptivity “ε”
and the path/cell length “b”. [4 ] A 6
ml aliquot of the sample was obtained
and diluted to 50 ml. 3 Absorbance
readings were taken using the same
UV-Vis Spectrophotometer [4]

Trial Absorbance Concentration (ppm)
1 0.138 169.7777778
2 0.139 170.8888889
3 0.139 170.8888889
Average 0.139 170.5185185
Table 2. Absorbance of Unknown
sample
Trial Concentration of Unknown Sol'n (ppm)
1 1415
2 1424
3 1424
Average 1421
Table 3. Cu (II) Concentration of
Unknown Solution
The calculated value of concentration
of the Unknown Solution gives us a
13.68% deviation from the theoretical
value of 1250 ppm, as disclosed by the
instructor. Analysis of the data of the
other groups was done and a pooled
standard deviation value of 0.524 was
obtained. This shows that the values
obtained experimentally were precise.
The shape of the calibration
curve often depends on the bandwidth.
That was why the wavelength range
needed to be scanned to determine
the best analytical wavelength for
analysis. At this wavelength, change in
absorbance with concentration is at a
maximum, thus yielding greater
sensitivity and higher accuracy.
Second, molar absorptivity is constant
at this band.[3]
However, Beer’s law is also
subject to limitations. First, deviations
3
become significant at higher
concentrations [4] since the refractive
index for the absorbed radiation is
changed at high concentrations. Thus
Beer’s law is ideally applicable to
solutions with concentrations below
1x10-2 M [3] that is why parts per
million (ppm) is used to indicate the
concentration of Cu(II). Also, Beer;s law
applies to as solution that may contain
more than one kind of absorbing
substance provided that there is no
interaction between them.[5] Chemical
deviations upon dilution are also
possible, leading to lesser actual
concentration in some substances like
the chromate ion.[2] Also, there are
also instrumental limitations to
consider: Stray radiation reaching the
detector, sensitivity changes and
power fluctuation. Fourth is when a
band of wavelengths is used rather
than monochromatic radiation [3] that
causes the detector to measure
average intensity instead of the
average of the log intensities. Another
source of error is the cuvette has
fingerprints, affecting the transmitted
radiation ang causing errors in the
Absorbance.
SUMMARY AND CONCLUSIONS
It could be said that Absorbance
is directly proportional to the
Concentration of the substance by
virtue of Beer’s law and was affirmed
by the experiment performed using a
UV-Vis Spectrophotometer. The
experiment was taken to be a success
given the small value of the Pooled
Standard Deviation for the experiment.
It was suggested that more accurate
results would be obtained if a quartz
cuvette was used.
REFERENCES
[1] Pickering, William F. 1966.
“Fundamental Principles of Chemical
Analysis”. Elsevier Publishing
Company.
[2] Klingenberg, Joseph. 1965.
“Introduction to Quantitative
Chemistry”. Reinhold Publishing Group
[3] Pecsok, Robert, et al. 1976.
“Modern Method of Chemical Analysis,
2nd edition”. John Wiley & Sons Inc.
[4] Bauman, Robert. 1962. “Absorption
Spectroscopy”. John Wiley & Sons Inc.
[5] Skoog, Douglas. 1962
“Fundamentals of Analytical
Chemistry” Holt, Rinehart and Winston,
Inc.
APPENDIX
A. Computation of Cu (II) ppm
ppm Std sol'n×Lsol'n÷Ltotal=ppm
Take L sol’n = 2 ml
4
2500mgL×0.002L.05L=100 ppm
B. Concentration (x) from linear
equation given absorbance (y)
x=y+yinterceptslopeppm
Take absorbance = 0.138
x=0.138+0.01480.0009 = 169.78 ppm
C. Average Concentration and
Absorbance
(Trial1+Trial2+Trial3)/3
169.78+170.89+170.893=170.52ppm
D. Concentration of Unkown
Solution
Concentration sample×Vdiluted
sol'nVolumeAliquot=ppm
Take Concentration Sample = 169.78
ppm
169.78 ppm×0.05L0.006L=1415ppm
E. Percent Deviation
Concentration-1250ppm1250×100%=
Take Concentration = 1421 ppm
1421-12501250×100=13.68%
D. Pooled Standard Deviation

Spooled=√((i=1n1Xi-X12+j=1n2Xi- V working Cu ppm Absorbanc

X22+k=1n3Xk-X32)/(n1+n2+n3+…- std mL e
ns))
2 100 0.082
0.823045267+0.823045267+09- 4 200 0.17
3=0.524
6 300 0.268

8 400 0.366

10 500 0.456

Sample Analysis

Aliquot 6
froms
Stock

Diluted 50
Unknown

Trial Absorba Concentra

nce tion (ppm)

1 0.138 169.77777
78

2 0.139 170.88888
89
RAW DATA
3 0.139 170.88888
GROUP A 89

Average 0.138666 170.51851

Team A 6 mL
667 85

Max λ 606.0 nm

Calibration Curve

Concentra 2500
tion ppm: GROUP B

5
Team B 11 ml

Average 0.259333 289.1481481

333
max 599
wavelengt
h

Calibration
Curve GROUP C

Concentra 2500
tion ppm:
Team C 14 mL

V working Cu ppm Absorbance

std mL Max λ 602.0 nm

2 100 0.093

4 200 0.18 Calibration

Curve
6 300 0.281
Concentratio 2500
8 400 0.365 n ppm:
10 500 0.458

V working Cu ppm Absorban

Sample std mL ce
Analysis 2 100 0.085
Aliquot 11 4 200 0.183
froms
Stock 6 300 0.273

Diluted 50 8 400 0.372

Unknown 10 500 0.457

Trial Absorba Concentrati Sample

nce on Analysis
1 0.26 289.8888889 Aliquot 14
2 0.259 288.7777778 froms Stock

3 0.259 288.7777778 Diluted 50

6
Unknown

Trial Absorban Concentr

ce ation

1 0.32 362.11111
11

2 0.32 362.11111
11

3 0.32 362.11111
11

Average 0.32 362.11111

11