University of Bahrain

College of Science
Department of Chemistry

Experiment 4
UV/Vis Spectrophotometry

Name : Mua'az Ahmed Taha

Academic Number : 20073459
Course Number : CHEMY 310
Submission Date : 2010-4-1

1
 Index

Page
o Objective………………………………….3

o Abstract…………………………………..3

o Theory…………………………………….3

o Requirements……………………………3

o Results……………………….…………..5

o Conclusions and Recommendations.....6

o References……………………………….6

2
Objective:
- To study the relation between the absorbance and the concentration for , Caffeine
and Asprin, using UV/Vis Spectrometry.
- To determine the concentration of a solution of Aspirin and Caffeine .
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 Aspirin and Caffeine
at a wave length of 273 nm and 300 nm , for different concentrations . for
instance , when the concentration was 30.4 ppm, wave length was 273 nm,
the absorbance was 1.283. From the obtain results we plotted the values
and found the slope for each curve at different wave lengths , for example :
at 273 nm for Aspirin , the slope was : 0.004. finally , from the slopes
obtained and the absorbance’s measured using the spectrophotometer for
the unknown concentration sample , the concentration is obtained to be ;
87.5249 ppm for aspirin at
Theory:

In chemistry, spectrophotometry is the quantifiable study of electromagnetic spectra. It is more

specific than the general term electromagnetic spectroscopy in that spectrophotometry deals with
visible light, near-ultraviolet, and near-infrared. Also, the term does not cover time-resolved
spectroscopic techniques.

Spectrophotometry involves the use of a spectrophotometer. A spectrophotometer is a photometer

(a device for measuring light intensity) that can measure intensity as a function of the color (or
more specifically the wavelength) of light. Important features of spectrophotometers are spectral
bandwidth and linear range of absorption measurement.

Perhaps the most common application of spectrophotometers is the measurement of light

absorption, but they can be designed to measure diffuse or specular reflectance. Strictly, even the
emission half of a luminescence instrument is a type of spectrophotometer.

The use of spectrophotometers is not limited to studies in physics. They are also commonly used in
other scientific fields such as chemistry, biochemistry, and molecular biology. They are widely used in
many industries including printing and forensic examination.

When a beam of radiation passes through a matter, a portion is frequently absorbed; this process
3
involves a transfer of radiant energy to the system, thus electrons of atoms or molecules are excited
to higher energy levels. In order for absorption to occur, the energy of the exciting photon must
match the energy difference between the ground state of the excited state of the absorbing species.

A study of the frequencies or wavelength of the absorbed radiation provides a mean of

characterizing the constituents of a sample mf matter. However, the quantity of the absorbed
radiation by certain species is a function of its concentration. The relationship between energy
absorption (Absorbance: A) and the concentration (C) of the absorbing species is given by:

A=εcl
Where :
- ε : the molar absorptivity
- l : the path length .

in practice , the quantity of the absorbed radiation is determined with a

spectrophotometer.

Absorption spectroscopy based upon ultraviolet and visible radiation is one of the most
useful tools available to the chemist for quantitative analysis. The important
characteristics of spectrophotometric and photometric methods are :

1- Wide applicability :
Enormous numbers of inorganic, organic , and bio-chemical species absorb
ultraviolet and visible radiation and are thus amenable to direct quantitative
determination .many nonabsorbing species can also be determined after chemical
conversion to absorbing derivatives. It has been estimated that over 90 % of the
analyses performed in clinical laboratories are based upon ultraviolet and visible
4
 absorption spectroscopy.

2- High Sensitivity: Typical detection limits for absorption spectroscopy range from 10 -
4
to 10-5 M .this range can often be extended to 10-6 or even 10-7 M with certain
procedural modifications.

3- Moderate to high Selectivity: often , a wave length can be found at which the
analyte along absorbs, thus making preliminary separations unnecessary.
Furthermore, where overlapping absorption bands to occur, corrections based upon
additional measurements at other wave lengths sometimes eliminate the need of
separation step .

4- Good Accuracy : the relative errors in concentration encountered with a typical

spectrophotmetric or photometric procedure employing ultraviolet and visible
radiation line in the range from 1 to 5 %. Such errors can often be decreased to a few
tenths of a percent with special precautions.

5- Ease and convenience : Spectrophotometric and photometric measurements are

easily and rapidly performed with modern instruments.in addition , the methods
readily lend themselves to automation.

Aspirin and caffeine are common components of analgesics. A two component

mixture may be analyzed by making absorbance measurements at two characteristic
maxima ( one for each component) , and solving the following pair of simultaneous
equations.

A1 = ε1c1l + ε2c2l at λ1 =273 nm

A2 = ε`1c1l + ε`2c2l at λ1 =300 nm
Where :
- ε: is the molar absorpitivity.
- λ: is the wave length.

Requirements:
- 25 ml Volumetric flasks.
- Pippettes.
- 250 ml Beaker.
- Measuring cylinders.

Data Collected:
Wave length (λ) = 273 nm Wave length (λ) = 300 nm
Caffeine Aspirin Caffeine Aspirin
Volume ppm Absorbance Volume Ppm Absorbance Volume ppm Absorbance Volume ppm Absorbance
5
2 30.4 1.283 1 60 0.296 2 30.4 0.009 1 60 1.076
4 60.8 2.323 2 120 0.552 4 60.8 0.025 2 120 1.530
6 91.2 2.41 3 180 0.781 6 91.2 0.046 3 180 2.04
8 121.6 2.44 4 240 1.061 8 121.6 0.074 4 240 2.28

Unknown mixture absorbencies:

- At (λ = 273) = 1.156
- At (λ = 300) = 0.573

Results:

For Caffeine:

6
For Aspirin :

7
Summary :

Aspirin Caffeine
Wave Length 273 nm 300 nm 273 nm 300 nm
Slope 2450 135 613 1030

Analysis for unknown mixture :

For λ = 273 nm ,
ε1c1(aspirin) + ε2c2(caffeine)
A1 =
1.156 =2450C1 + 135C2 -(1)
For λ = 300 nm ,
A2 =ε’1c1(aspirin) + ε’2c2(caffeine)
0.573 =613C1 + 1030C2 –(2)

Then by solving equations (1) and (2) , simultaneously ;

C1 = 4.56 x 10-4 M aspirin (mol/L)---(MM = 180 g/mol) ----C1=0.08208

M(g/mol)
C2 = 2.85 x 10-4 M .

Conclusion and Recommendation:

References:
1- http://en.wikipedia.org/wiki/titration
2- SKOOG/WEST/HOLLER, Fundamentals Of Analytical Chemistry, Saunders HBJ ,
6th Edition, 1992.