NAME

SEGOKGO

STUDENT NUMBER

GROUP/DAY

: LORRAINE KESEGO

: 201002660

: TUESDAY

WEEK

: THIRD

DATE

: 12/MARCH/2013

TITLE: THE SIMULTANEOUS DETERMINATION OF

CAFFEINE AND ACETYLSALICYLIC ACID IN AN
ANALGESIC BY ULTRAVIOLET SPECTROPHOTOMETRY

AIM
To figure out the amount concentration of caffeine and acetylsalicylic acid in
an analgesic tablet by ultraviolet spectrophotometry.
INTRODUCTION
In this experiment, you will determine the amount of caffeine and
acetylsalicylic acid in an analgesic tablet by UV spectrophotometry. Many
molecules absorb ultraviolet or visible light. When an atom or molecule
absorbs energy, electrons are promoted from their ground state to an excited
state. Absorption spectrometry involves measuring the fraction of light of a
given wavelength that passes through a sample. When a monochromatic
light beam passes through a layer of solution with a thickness, absorptivity
and concentration of an absorbing species, as the consequence of
interactions between the photons and absorbing particles, the power of the
beam is attenuated from Po to P. The absorbance A of a solution is defined by
the equation:
A = log (P / Po) = abc;
This is Beers law, where a is a proportionality constant called absorptivity
and b is the path length of the light beam through the absorbing medium.
When the c is expressed in M (moles per liter), and b in cm, a is called the
molar absorptivity and is given the special symbol , with the units of L cm-1
mol-1. Thus,
A = bc
In this experiment, there are two components caffeine and acetylsalicylic
acid) in the analgesic tablet. You will first determine the molar absorptivity
of each component by constructing a calibration curve (absorbance vs.
concentration) with standard solutions. Then by measuring absorbance of
the tablet solution at maximum absorption wavelength of both components,
you will be able to figure out the amount of each component in the tablet.
UV-vis Spectrophotometer Systems utilize the optical bench of Ocean Optics
second-generation miniature fiber optic spectrometer, the S2000, by
mounting it onto an A/D converter and turning the system into a PC plug-in
spectrometer. The UV-vis consists of four basic elements: the PC2000 UV-vis
PC Plug-in Fiber Optic Spectrometer (200-850 nm), a miniature deuterium
tungsten light source with integrated cuvette holder, a 300-m solarization
resistant optical fiber, and OOIChem operating software. The light source
supplies light to the sample. The light transmitted through the sample is

collected and sent to the spectrometer via the fiber. The spectrometer
measures the amount of light at each wavelength in the sampled spectrum.
The A/D converter, on which the spectrometer is mounted, transforms the
analog data from the spectrometer into digital information that is passed to a
computer. Finally, the software performs basic acquisition and display
functions on your data.
PRELABORATORY ASSIGNMENT

EXPERIMENTAL SECTION
APPARATUS

REAGENTS

2* 1-cm curvets
1-ml pipette
2-ml pipette
Scanning UV-Vis spectrophotometer
14* 50ml volumetric flasks
3* 250ml volumetric flasks

Methanol
Caffeine
Acetylsalicylic acid
Analgesic tablet

a) REAGENT AND APPARATUS

b) PROCEDURE
0.117g of caffeine and 0.12g of acetylsalicylic were weighed and dissolved
into methanol. The solutions were then transferred into labeled 250-ml
volumetric flasks then filled to the mark with methanol. From the caffeine
solution, five different standards of 0.25, 0.35, 0.45, 0.55 and 0.65 ml were
prepared in labeled 50-ml volumetric flasks and they were filled to the mark
by methanol. The same procedure was also followed to prepare five
standards of acetylsalicylic and the volumetric flasks were labeled. The
pipettes were used to deliver 0.25-ml and 0.35-ml of caffeine and
acetylsalicylic acid in a 50-ml volumetric flask filled to the mark by methanol.
0.045g of analgesic tablet was weighed and dissolved with methanol. The
solution was then transferred into a 100-ml volumetric flask then filled to the
mark by methanol. 1.0-ml of the sample solution was added to each 50-ml
volumetric flasks then filled to the mark by methanol. The UV-Vis
spectrophotometer was the adjusted by setting the initial wavelength as
220nm and final as 320nm. The two curvets were used and the other had
methanol and another was used to measure the blank. The baseline
absorbance was set to zero and the other curvets were used to measure
different standards of caffeine and acetylsalicylic acid and also the sample.

RESULTS AND ANALYSIS OF DATA

From the stock solution; mass of Caffeine measured= 0.1174g & molar mass
is 194.17g/mol
Moles of Caffeine= 0.1174g/194.19g.mol-1 = 0.0006046moles
[Caffeine]= 0.0006046moles/250*10-3L = 0.002481M
Concentration of Caffeine= 2.418*10-3 M
For the standards; [caffeine] = (2.418*10-3 M*50ml)/250ml
= 4.836*10-4 M
From the stock solution; mass of acetylsalicylic acid measured is 0.1239g &
molar mass is 180.15g/mol
Moles of Caffeine= 0.1239g/180.15 g.mol-1 = 0.0006878moles
[Acetylsalicylic acid]= 0.0006878moles/250*10-3L = 0.002751M
Concentration of acetylsalicylic acid = 2.751*10-3 M
For the standards; [Acetylsalicylic acid] = (2.751*10-3 M*50ml)/250ml
= 5.502*10-4 M
The table for the % absorbance against concentration of caffeine which was
270nm
CAFFEINE
-5
CONC 10
ABS
FACTOR
CORRECTE
D
STD1
1.25
0.646
0.522
0.124
STD2
1.68
0.681
0.522
0.159
STD3
2.16
0.686
0.522
0.164
STD4
2.64
0.761
0.522
0.239
STD5
3.12
0.804
0.522
0.282
BLANK
0
0.522
SAMPLE
0.584
MIXTUR
0.663
E
The graph shows that the sample had 0.7*10-5 M of caffeine
The table for the % absorbance against concentration of acetylsalicylic acid
which was 225nm
ACETYSALICYLIC ACID
CONC 10-5
ABS
FACTOR
CORRECTE
D
STD1
1.35
3.914
3.587
0.327
STD2
1.89
3.419
3.587
-0.168
STD3
2.43
3.738
3.587
0.151

STD4
STD5
BLANK
SAMPLE
MIXTUR
E
The graph
acid.

2.97
3.51
0

3.738
3.914
3.587
3.587
3.437

3.587
3.587

0.151
0.327

shows that the sample had around 0.04*10-5 M of acetylsalicylic

GRAPHS;
a)

THE GRAPH OF ABSORBANCE AGAINST CONCENTRATION OF CAFFEINE TO DETERMINE THE CONCENTRATION OF THE SAMPLE
0.9
0.8

f(x) = 0.09x + 0.53

R = 0.98

0.7
0.6
0.5
%Absorbance 0.4
0.3
0.2
0.1
0
0

0.5

1.5

Concentration/ 10-5 M

b)

2.5

3.5

THE GRAPH OF % ABSORBANCE AGAINST CONCENTRATION OF ACETYSALICYLIC ACID TO DETERMINE THE CONCENTRATION OF SAMPLE
4
3.9
3.8

f(x) = 0.06x + 3.59

3.7 R = 0.17
3.6
% Absorbance 3.5
3.4
3.3
3.2
3.1
0

0.5

1.5

2.5

3.5

Concentration/10-5 M

DISCUSSION
The results obtained shows that there is 0.7*10-5 M of caffeine at the
wavelength of 270nm and 0.004*10-5 M of acetylsalicylic acid at the
wavelength of 225nm. The values obtained for concentration and %
absorbance of caffeine seem to be prcised because of the r-squared value
which was 0.9777 which is close to 1 whereas the values for acetylsalicylic
acid were not at all prcised because the r-squared value was 0.1673 which
far from 1. This might have been caused by random error because when
making standard solutions, some of the solution was lost through a pipette
making the solution to be less than the required amount and also during the
dilution of both the stock solutions and the standards especially for
acetylsalicylic acid because the solvent which is methanol was poured
beyond the mark.

CONCLUSION
The analgesic tablet had 0.7*10-5 M of caffeine at the wavelength of 270nm
and 0.004*10-5 M of acetylsalicylic acid at the wavelength of 225nm.

REFERENCES
1. Pfandl A., Quantitative analysis of pharmaceutical preparations
containing analgesic and antipyretic agents, 108 (1968) 568-571.
2. Turkish J., Simultaneous determination of active ingredients in
binary mixtures containing caffeine using liquid chromatographic
and spectrophotometric methods, 2 (2004)115-138.