Experiment Title: Quantitative Analysis of Aspirin Tablets by Double

Beam Ultraviolet Absorption Spectrophotometry

Subject: UEMK2023

Course: Chemical Engineering

Group: G4

Name of Lecturer: Dr. Ng Yee Sern

Name of Student Student ID No Year and Semester

Chung Wen Loong 1507063 Y2S3
Loo Wei Ting 1507760 Y2S3
Ong Kha Kuan 1507378 Y2S3
Nyoi Kai Heng 1502166 Y2S1
Tan Kian Hock 1506732 Y2S3

Received by: ____________________ Date: ________________

Lab Asst/ Lab Officer

Lecturer

Receipt of Lab Report Submission

(To be keep by student)
Experiment Title: ________________________________________

Subject: ________________________________________

Course: ________________________________________

Name of Student Student ID No Year and Semester

Chung Wen Loong 1507063 Y2S3
Loo Wei Ting 1507760 Y2S3
Ong Kha Kuan 1507378 Y2S3
Nyoi Kai Heng 1502166 Y2S1
Tan Kian Hock 1506732 Y2S3

Received by: ____________________ Date: ________________

Lab Asst/ Lab Officer

Title: Quantitative Analysis of Aspirin Tablets by Double-Beam Ultraviolet Absorption
Spectrophotometry.

Objectives:
1. To practice the instrumental analysis techniques by preparing standard solution.
2. To determine the concentration of the salicylic acid from the photon absorption by using
Beers Law.
3. To determine the amount of aspirin (acetylsalicylic acid) in commercial analgesics.

Introduction:
In 1899, the Acetylsalicylic acid (ASA) was introduced under the trademark- Aspirin. ASA
are function as a treatment on pain, inflammation and fever. However, a huge amount of ASA in
aspirin tablets will cause bad consequences on human body. Therefore, the quantitative analysis of
aspirin tablets was introduced. The purpose of the quantitative analysis of aspirin is to determine the
concentration of acetylsalicylic acid (ASA) in aspirin tablets (JENWAY).
In pharmaceutical analysis field, the method- Ultraviolet-Visible spectrophotometry is most
often used to determine the visible radiation or the amount of ultraviolet absorbed by a substance in
a solution. The Ultraviolet-Visible spectrophotometers is the instrument that determine the ratio of
the intensity of double beam of light source in the U.V-Visible region. There are 2 analysis which is
qualitative and quantitative analysis. Organic compound can be determined through qualitative
analysis by using the spectrophotometry. On the other hand, the quantitative spectrophotometry
analysis can determine the amount of absorption of electromagnetic radiation by molecules species.
This quantitative spectrophotometric analysis is governs by Beer-Lambert law.

FUNDAMENTAL BEERs LAMBERTs

LAW LAW LAW
The intensity of a With the number of absorbing As it passes through a medium of
beam of parallel molecules. Homogenous thickness.
monochromatic
radiation reduce
exponentially

Beer-Lamberts Law state that When beam of light is passed through a transparent cell
containing a solution of an absorbing substance, reduction of intensity of light may occur. (Behera,
2012) Mathematically, Beer-Lamberts Law is defined as:
A= bc
where,
A = absorbance or optical density
a = absorptivity or extinction coefficient
b = path length of radiation through sample (cm)
c = concentration of solute in solution
Both b and are constant so A is directly proportional to the concentration c.

Experimental Details:
The Varian Cary 100 UV-Visible Double Beam Scanning spectrophotometer was turned on for 15
minutes.
Preparing Standard Solution
1) 0.1g (to 0.1mg) of pure salicylic acid was weighed and transferred to a 100cm3 volumetric
flask.
2) 0.1 mol dm3 of NaOH solution was prepared after mixing and dilution.
3) The salicylic acid concentration, in mg dm-3, of the diluted standard solutions was calculated.
4) A series of five diluted solution from solution was prepared by transferred 1.00, 2.00, 3.00,
4.00 and 5.00 cm3 of stock solutions into a series of 5 100- cm3 volumetric flasks and the
concentration of the solution was labeled.
5) The salicylic acid concentration of the diluted standard solutions, in mg dm-3 was calculated.
Calibration Curve
1) The 3.00 cm3 of diluted solution of salicylic solution was scanned and the UV absorption
spectra was obtained by using Varian Cary 100 UV-Visible Double Beam Scanning
spectrophotometer.
2) The wavelength of the maximum absorbance was computed.
3) The absorbance for each of the standards at the wavelength of maximum absorbance was
measured.
4) The calibration curve for the salicylic acid standards with the measured absorbance
(Absorbance vs Concentration) was plotted.
Aspirin Analysis
1) One commercial aspirin tablet was weighed and dissolved in 0.1 dm-3 NaOH solution in a
250-cm3 volumetric flask.
2) The volumetric flask was mixed thoroughly by shaking the volumetric flask.
3) Sample solution was diluted with 0.1 mol dm-3 NaOH solution to ratio of 1: 50 and the
absorbance of this solution at the same wavelength was measured.
 4) The calibration curve was used and the concentration of salicylic acid present in the sample
solution was found.
5) The weight of acetylsalicylic acid (ASA) in the aspirin tablet that apply the appropriate
stoichiometric factors was computed. ( consider the dilutions involved)
6) The results to the content specified on the label was compared.
Unknown Analysis
1) By using the calibration curves from the scanning UV-VIS, concentration of salicylic acid in
the unknown solution, Y (after appropriate dilutions) was determined.

Reagents:
0.1 mol dm-3 NaOH
Salicylic acid
Commercial aspirin tablets
Unknown solution, Y

Apparatus:
Burette, 25.00 cm3 with graduation in 0.05 cm3 divisions
100-cm3 and 250-cm3 volumetric flasks
250-cm3 and 500-cm3 conical flasks
Analytical balance
UV-Vis double-beam scanning spectrophotometer

Results and Calculation:

Formula:

Mstd Vstd = Mstock Vstock

where,

Mstock= Initial concentration/ molarity of salicylic acid/aspirin (acetylsalicylic acid) before diluted

Vstock= Initial volume of stock solution before diluted

Mstd =Final concentration of salicylic acid/aspirin (acetylsalicylic acid) after diluted

Vstd = final volume of stock solution after diluted (100 cm3 in volumetric flasks)
Calculation of the concentration/ molarity of the salicylic acid stock solution,

Mass of pure salicylic acid

Mstock =
Volume of solvent
0.1g 1000mg 1cm3
Mstock =
100cm3 g 0.01 L
mg
Mstock = 1000
L

At 1cm3 of the stock solution:

Mstd Vstd = Mstock Vstock

Mstock Vstock
Mstd =
Vstd
mg
(1000 L ) (1cm3 )
Mstd =
(100cm3 )
mg
Mstd = 10
L
1
= 10 138.121
1000

= 7.24003 105

At 2cm3 of the stock solution:

Mstd Vstd = Mstock Vstock

Mstock Vstock
Mstd =
Vstd
mg
(1000 L ) (2cm3 )
Mstd =
(100cm3 )
mg
Mstd = 20
L
1
= 20 138.121
1000

= 1.448 104
At 3cm3 of the stock solution:

Mstd Vstd = Mstock Vstock

Mstock Vstock
Mstd =
Vstd
mg
(1000 L ) (3cm3 )
Mstd =
(100cm3 )
mg
Mstd = 30
L
1
= 30 138.121
1000

= 2.172 104

At 4cm3 of the stock solution:

Mstd Vstd = Mstock Vstock

Mstock Vstock
Mstd =
Vstd
mg
(1000 L ) (4cm3 )
Mstd =
(100cm3 )
mg
Mstd = 40
L
1
= 40 138.121
1000

= 2.896 104

At 5cm3 of the stock solution:

Mstd Vstd = Mstock Vstock

Mstock Vstock
Mstd =
Vstd
mg
(1000 L ) (5cm3 )
Mstd =
(100cm3 )
mg
Mstd = 50
L
1
= 50 138.121
1000

= 3.620 104
 Concentration of standard solution Absorbance
Standard solution
(mol/L)
1 0.000072400 0.310
2 0.000144801 0.515
3 0.000217201 0.732
4 0.000289601 1.011
5 0.000362001 1.266

Graph of Absorbance against Concentration

1.400
y = 3326x + 0.0444
1.200 R = 0.9961
1.000
Absorbance

0.800

0.600

0.400

0.200

0.000
0 0.00005 0.0001 0.00015 0.0002 0.00025 0.0003 0.00035 0.0004
Concentration (mol/L)

Graph of Absorbance against Concentration (Calibration Curve)

From the calibration curve above, the concentration of the 1:10 aspirin in the sample can be calculated.
Absorbance of the 1:10 aspirin in sample = 0.581
= 3326 + 0.0444
0.0444
=
3326
0.581 0.0444
=
3326

= 1.613 x 104

Mass of aspirin in sample in 250cm3,

1.613 x 104 1 180.157 1000

= 250 3
10003 1
= 7.265

Mass of aspirin in the tablet before dilution,

= 7.265 10

= 72.65
The content of ASA specified on the label is 100mg. So the percentage error can be calculated by:

= | | 100%

72.65100
=| | 100%
100

= 27.35%

Discussion:

The experiment that we conducted will test out the concentration of aspirin from different
samples (acetylsalicylic acid) by using Double-Beam Ultraviolet Absorption Spectrophotometry.
When the sample molecule is exposed to incoming light with energy which has the similar electronic
transition within the molecule, some of the light will be absorbed as the electron is promoted to high
energy orbital (from a full orbital into an empty anti-bonding orbital). Energy from the light will be
used to promote an electron from a bonding or non-bonding orbital into one of the empty anti-bonding
orbitals.

E = hv =

Based on the formula above, the higher the energy jump, the higher the frequency of light to
be absorbed. However, UV-visible Absorption Spectrophotometry always use wavelength instead of
frequency. Therefore, the larger the gap for energy to jump, the lower the wavelength of the light is
being absorbed. Hence, we can conclude that aspirin has a bigger energy gap which causing it to
absorb a lower wavelength light -297nm (refer to appendix figure 1.A) and it falls within the
colourless wavelength range (200nm-350nm)
According to Beers Law, A= bc , the amount of light being absorbed by a sample is
dependent on the path length (b), concentration of the sample (c) and a proportionality constant (-
molar absorptivity). As the concentration of aspirin increases, the more light will be absorbed by the
sample, hence the higher the absorbance level. Absorbance usually ranges from 0 (no absorption) to
2 (99% absorption), and is precisely defined in context with spectrometer operation. Based on the
graph that we obtained, sample 5 (1.266) has the highest value of absorbance followed by sample 4
(1.011), sample 3 (0.732), sample 2 (0.511), and sample 1 (0.310). Besides that, the fluctuation
caused before 297nm is considered as noise due to the presence of impurities. In short, when the
concentration of aspirin increases, there will be more molecules to impede the photons. Hence, this
will cause a higher concentration aspirin seem darker and providing it a higher absorbance.
In the experiment, the final salicylic acid concentration and absorbance of 5cm3 of the diluted
standard solution is the highest value which are 0.000362001 mol/L and 1.266 respectively whereas
the lowest value of the final concentration and absorbance is the dilution factor = 1:50 of 1 mg/L
sample solution (aspirin) which only have 2.39327 X 10-5 mol/L and 0.124 respectively compared
to other diluted standard solutions. 100 UV-Vis double-beam scanning spectrophotometer is used to
determine the amount of light of a specific wavelength absorbed by an analyte. The concentration of
the salicylic acid is directly proportional to the absorbance based on the formula A= bc (Chasteen,
2009). After that, the specific wavelength (297nm) is chosen based on the absorbance characteristics
of the analyte which are the salicylic acid and acetylsalicylic acid (aspirin).
There are some errors that may cause the difference in reading between the experimental
value and the calculated value for the aspirin concentration in diluted standard solutions. There is
zero percentage error in final concentration of salicylic acid as we had prepared well in the
experiment and be avoided of other careless error occurred whereas there is a percentage error of
27.35% between experimental value and the labelled value of concentration of aspirin in sample.
This is because the dilution method we used by adding sodium hydroxide (NaOH) into the sample is
not quite accurate and the quartz cuvettes are not rinsed properly by using NaOH solution during the
experiment. Beside that, the outside wall of quartz cuvettes are not cleaning up properly before
placing in the UV-VIS spectrophotometer as well as the malfunction of equipment we used will be
affected the accurateness of the results needed.
Commercially, the amount of aspirin (acetylsalicylic acid) available in aspirin tablet can be
determined by the neutralization reaction where the aspirin is being hydrolysed to salicylic acid
before diluting to a concentration in which the Beers Law is obeyed via Atomic Spectroscopy
method (Jenway, 2014). There are some reasons on why is the salicylic acid stock solution prepared
using 0.1 mol dm-3 NaOH solution. The aspirin was dissolved using NaOH instead of water because
aspirin is a weak acid, hence it only dissolves partially in water. In facts, aspirin is more soluble in
alkaline solutions such as NaOH. Thus, the aspirin will dissolve completely in sodium hydroxide and
will be converted to its sodium salt which is soluble in water to find its concentration (Ausetute,
N.D). Besides, the hydrolysis of aspirin occurs more swiftly in alkaline condition. Aspirin is the
acetate ester of salicylic acid,2-hydroxybenzoic acid. During hydrolysis, the acetyl ester breaks down
rapidly to the salicylate dianion and acetate anion in the basic medium. The dianion of aspirin are
more soluble in water as there are strong ion-dipole interactions with H2O molecules. Hence, by
adding water, the salicylate dianion will be converted into the salicylic acid (Prenzler, 2006). The
hydrolysis of aspirin is shown below:
(Jenway, 2014)

In facts, there are differences between aspirin and also salicylic acid. The chemical formula
of aspirin is C6H4OCOCH3COOH while for salicylic acid, its chemical formula is C6H4OHCOOH
(CR Scientific LLC, N.D). In term of molecular structure, aspirin is actually both an aromatic acid
and an ester. It consists of carboxylic acid functional group (R-COOH), ester functional group (R-O-
CO-R) and also aromatic group which is the benzene ring. As for salicylic acid, it is a phenolic acid
which consists of carboxylic acid functional group (R-COOH), hydroxyl functional group (R-OH)
and also aromatic benzene ring. Actually both salicylic acid and aspirin are interrelated. Salicylic
acid is the mother compound of aspirin. In the other words, the hydrolysis of aspirin will produce
salicylic acid while aspirin can be formed via the esterification of the phenolic hydroxyl group of
salicylic acid (Ausetute, N.D). The preparation of salicylic acid stock solution can be done by using
either pure aspirin or pure salicylic acid. In the experiment, 0.1g of pure salicylic acid is used. The
molecular weight of salicylic acid is 138.121g/mol (Helmenstine, 2017). By dividing 0.1g with its
molecular weight, the amount of salicylic acid used is 0.000724 moles. In the hydrolysis process, for
every mole of salicylic acid to be produced, 1 mole of aspirin was needed. Thus, the mole ratio
between them is 1. Hence, the amount of aspirin needed is also 0.000724 moles. The molecular
weight of aspirin is 180.157g/mol (ConvertUnits.Com, 2017). By multiplying the molecular weight
of aspirin with 0.000724 moles, we will obtain the weight of aspirin which is 0.1304g. In short, if
aspirin is used to prepare the desired salicylic acid stock solution, its weight needed will be 0,1304g.
The measure that should be taken due to safety is wearing a pair of gloves during conducting the
experiment. It is because sodium hydroxide solution is used during the experiment, sodium hydroxide
is corrosive and may burn your hands. Place the eyes level at the same position to the meniscus during
measuring the volume of solution needed to avoid parallax error otherwise, there will be an error in
the reading. Shake the volumetric flasks which contain the mixture of salicylic acid and sodium
hydroxide or aspirin and sodium hydroxide to ensure the concentration of the solution is correct. Use
different droppers for each solution and rinse the quartz cuvettes with sodium hydroxide for each
experiment to avoid the contamination of components between mixtures. Clean the outside wall of
quartz cuvettes before placing it into UV-VIS spectrophotometer to obtain accurate values of
absorbance. The level of solution placed in the quartz cuvettes should be about 80% otherwise the
spectrophotometer cannot detect its concentration if the level is too low or the sample will spill if the
level is too high.
There are some remarks for group 1. Group 1 has great organization and good time management
during conducting the experiment. The preparations and procedures were going well except the
testing of the absorbance of 50mg/L salicylic acid, the Group 1 was not cleaning the wall of quartz
cuvette properly before placing it into the UV-VIS spectrophotometer and resulted in the wrong
fluctuation on the wavelength-absorbance graph.

Conclusion:
As a conclusion of this experiment, we can summarize that the absorbance of light is directly
proportional to the concentration of salicylic acid. The higher the concentration of salicylic acid, the
higher the absorbance of light. Moreover, we concluded that the 1: 50 aspirin in NaOH solution is
too diluted, and it does not fulfill the Beers Law principles. After a few appropriate dilution, we
decided to use 1: 10 aspirin in NaOH solution. As result, the 1: 10 aspirin in NaOH solution fulfilled
the Beers Law principles. After doing the experiment, we conclude that the experiment value of the
mass of aspirin in sample and the labelled value have a percentage error of 27.35% which might be
for safety reason or experimental errors. Minor errors may lead to huge effect to outcome, causing
imperfect and inappropriate results to be obtained.
References:

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http://www.ausetute.com.au/aspirin.html

2. Behera, S. (2012, October 31). Analytical & Bioanalytical Techniques. Retrieved from UV-Visible
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5. ConvertUnits.Com. (2017). Molecular Weight of Aspirin. Retrieved June 25, 2017, from ConvertUnits.com:
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