Department of Chemistry

University of Bahrain
CHEMY310

Experiment.4
UV/VIS
Spectrophotometry

Name: Zahra Ali Isa Albasri

ID: 20165036
Sec: 02
Abstract:
The UV/Vis spectrometry is powerful technique, that is used in several fields
such as : medicine , forensic . in this experiment we used a spectrophotometer
to determine the absorbance for Cobalt and Nickel at a wave length of 510 nm
and 395 nm , for different concentrations .

Introduction :
Spectrophotometry is an analytical technique which uses light to measure the
chemical concentrations of certain samples. The spectrophotometer will
measure the amount of photons (intensity of the light) absorbed after it passes
through the sample solution. The concentration of the sample can be
determined by measuring the intensity of the light detected. In general, a
spectrophotometer consists of two devices; a spectrometer and a photometer.
First, the spectrometer produces the wavelength for the desired range in the
experiment. The lens transmits a straight beam of light (photons) that will pass
through a prism and split into several wavelengths. Then a slit transmits only
the desired wavelengths. Secondly, the desired wavelengths pass through the
solution in the cuvette and the photometer will detect the amount of photons
and send the signal to a galvanometer or a digital display. For this
experiment, UV-visible spectrophotometry will be used to
determine the concentrations of cobalt and nickel in an unknown
solution. Light will be used are (395 and 510nm) of electromagnetic
radiation spectrum. In spectrophotometry, absorbance of the sample is
dependent and directly proportional to the concentration of the absorbing
species. This can be related with the Beer-Lambert Law: A=ε L c where ε is the
molar absorptivity for the absorbing species, L is the path length (the distance
the light passes through the absorbing species), and c is the concentration of the
absorbing species.

Materials and Method:

25ml volumetric flasks, pipettes, 100ml beakers, measuring cylinders
CO(NO3)2(0.15M) and Ni(NO3)2 (0.15M) .
UV Spectrophotometry is the method used.
For procedure refer to lab manual .
Results and calculations

Volume Calculations:

M x V = M2 x V2

M2 ∗ V2
V=
M

(0.025) ∗ (10)
V1 = = 1.6𝑚𝑙
(0.15)

(0.0375) ∗ (10)
V2 = = 2.5𝑚𝑙
(0.15)

(0.075) ∗ (10)
V3 = = 5.0𝑚𝑙
(0.15)

(0.1125) ∗ (10)
V4 = = 7.5𝑚𝑙
(0.15)
ƛ=395nm

Concentration absorbanceof absorbance

no. Volume (M) Cobalt of Nickel
1 1.6 ml 0.025 -0.009 0.118
2 2.5ml 0.0375 -0.004 0.192
3 5.0ml 0.075 0.007 0.685
4 7.5ml 0.1125 0.04 0.78
Absorbance vs.Concentration y = 0.5387x - 0.0252
cobalt 395nm R² = 0.9335

0.06
Absorbance nm

0.04
0.02
0
-0.02 0 0.02 0.04 0.06 0.08 0.1 0.12
Concentration M

Absorbance vs.Concentration y = 8.1787x - 0.0674

Nickel 395nm R² = 0.9202

1
Absorbance nm

0.8
0.6
0.4
0.2
0
0 0.02 0.04 0.06 0.08 0.1 0.12
Concentration M

For ƛ=510nm

Concentration Absorbance absorbance

no. Volume (M) of Cobalt of Nickel
1 1.6 ml 0.025 0.112 -0.001
2 2.5ml 0.0375 0.166 0.008
3 5.0ml 0.075 0.341 0.306
4 7.5ml 0.1125 0.543 0.399
 Absorbance vs.Concentration y = 4.92x - 0.017
cobalt 510nm R² = 0.9982

0.6
Absorbance nm

0.5
0.4
0.3
0.2
0.1
0
0 0.02 0.04 0.06 0.08 0.1 0.12
Concentration M

Absorbance vs.Concentration
y = 5.0373x - 0.1368
Nickel 510nm R² = 0.9428
0.5
0.4
Absorbance nm

0.3
0.2
0.1
0
-0.1 0 0.02 0.04 0.06 0.08 0.1 0.12
Concentration M

For unknown no. 1 Absorbance @395nm = 0.147 and @510nm=0.243

From Graphs of Cobalt :

Slope of Graph @ 395nm = ε1 = 0.5387
Slope of Graph @ 510 nm = ε2 = 4.9
From Graphs of Caffeine
Slope of Graph @ 395nm = ε1’= 8.1787
Slope of Graph @ 510nm = ε2’ = 5.0373
A1 = ε1C1L + ε1’ C2L @ 395nm
A2 = ε2 C1L + ε2’C2L @ 510nm

0.147 = 0.5387C1+8.1787C2 …….>Eq1

0.243= 4.9C1+5.0373 C2 …….>Eq2

0.147– 8.1787 𝐶2
𝐶1 =
0.5387
Substitute value of C1 at eq2
Using the solver in calculator,

C2 = 0.0157M
C1= 0.1635 M

Discussion:
In this experiment, we determined the absorbance of least concentrated
caffeine and aspirin using UV-spectrophotometer and it was found to be
395nm and 510nm hence we set our spectrophotometer on these both
wavelengths to complete of the determination of the remaining
concentrations, then we plot the relation between absorbance and
concentrations ,and from this plot we could determine the concentration of
unknown mixture containing specific percentage of aspirin and caffeine by
using equation (1)&(2) .
The correction factor R2 = 0.92 ̴ 0.99 which that means there is no errors in
the experiment .

Conclusion:
The absorbance was taken for tow wave lengths, 395nm and 510nm, and the
concentration founded for C1 and C2 were as below:
Concentration of Cobalt, C1= 0.1635 M
Concentration of Nickel, C2 = 0.0157M
Also the “epsilon” values were:
ε1 = 0.5387, ε2 = 4.9, ε1’= 8.1787, ε2’ = 5.0373 .

Reference:
 - http://www.tandfonline.com/doi/abs/10.1080/00032710903486260
 LJ Rogers, Simultaneous determination of paracetamol, aspirin and
caffeine in tablet formulations using factor analysis, Journal of Analytical
Communications,1996.33,401-402.