UNIVERSITI TUNKU ABDUL RAHMAN

BACHELOR OF ENGINEERING (HONS)

CHEMICAL ENGINEERING

UEMK2023

INSTRUMENTAL ANALYSIS

Lab Report

Lecturer: Dr. Ng Yee Sern

Date of Submission: 21 June 2017

Tutorial Course Year

Name ID Number
Group
Fong Jia Qi 1506851 5 CL Y2S3

Ching San 1203458 1 CL Y4S3

Tee Yi Ting 1405002 5 CL Y2S2

Yong Xin Ni 1506991 5 CL Y2S3

Title

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

Objectives

1. Construct calibration curve based on Beers Law.

2. Use Beers Law to determine molar absorptivity.
3. Explain the fundamental principal behind spectrophotometric analysis.

Introduction

Aspirin (acetylsalicylic acid) is a salicylate drug which important to be used for its analgesic,
antipyretic and anti-inflammatory properties. Aspirin is a white, crystalline, weakly acidic substance
which melts at 135C. It is one of the most widely used medications as a pain reliever.

Figure 1: Preparation of Aspirin from Salicylic Acid

Electromagnetic radiation absorption by molecules form basis for a multitude of instrumental

analytical techniques. Beers Law can determine the analyte concentration from photon absorption
with wavelengths in ultraviolet range.

Direct calibration curve method can be applied for analyzing unknown sample only when standard
solutions and unknown solution are prepared and measured under exactly the same conditions.
However, substances other than analyte will affect absorbance reading of sample solution at
wavelength chosen for analysis, known as matrix effects. This can be avoid by conducting the standard
addition method so that the matrix present in the sample can affect the absorbance readings for both
the analyte in the standard and the sample. Therefore, it is possible to analyze the analyte accurately
even in the presence of matrix.

Method of standard additions can be used to improve the result in certain circumstances the matrix,
which contributes significantly to the absorbance of a sample and is also highly variable. The basic
idea is to add standard to analyte sample so that the standard is subjected to the same matrix effects as
analyte, this method assumes that the system obeys the Beers Law. Aspirin (acetylsalicylic acid)
amount in commercial analgesics can be determined when it has been hydrolyzed to salicylic acid and
diluted to a concentration where the law is obeyed.
Reagents and Apparatus

Item Description Item Category Quantity

UV-VIS spectrophotometer Equipment 1
Analytical Balance Equipment 1
Salicylic acid
Chemical 1 set
(5 standard solutions)
Sodium Hydroxide Chemical 500mL
Quartz Cuvettes Glassware 1
Beaker (100mL) Glassware 3
Water Dispenser Tool 1
Spatula Tool 1
Distilled Water Consumable 2 liter
Aspirin Tablet Consumable 1
Tissue (Kim wide) Consumable 1
Droppers Consumable 5
Results and Graph

Standard Solution

Solution Volume (cm3) Concentration (mg/L) Absorbance

1 1.00 10 0.278
2 2.00 20 0.553
3 3.00 30 0.831
4 4.00 40 1.064
5 5.00 50 1.233

Graph of Absorbance against Concentration

1.4

1.2
y = 0.0242x + 0.0655
1 R = 0.9912
Absorbance

0.8

0.6 Absorbance

0.4 Linear (Absorbance)

0.2

0
0 10 20 30 40 50 60
Concentration(mg/L)
Sample Calculations

For M1,

1000mg
0.1g(1g )
M1 =
0.1L
100cm3 ( )
100cm3

M1 = 1000mg/L

For volume of 1.00cm3,

M1 V1 = M2 V2

M1 V1
M2 =
V2

0.1L
(1000mg/L)(1.00cm3 )( )
M2 = 100cm3
0.1L
100cm3 ( )
100cm3

M2 = 10mg/L

The calculation is repeated with different volume of standard solutions.

Aspirin Solution

Ratio of salicylic acid to NaOH Absorbance Concentration (mg/L) Weight (mg)

1:10 0.114 2.00 0.5
1:50 0.553 20.14 5.035

Sample Calculations

At volume of 10cm3,

Based on the equation obtained at the graph,

y = 0.0242x + 0.0655

Where y = Y = Absorbance, m = 0.0242, x = X = Concentration of salicyclic acid and c = 0.0655

 y 0.0655
x=
0.0242

When y = 0.114,

0.114 0.0655
x=
0.0242

x = 2.00mg/L

For the weight of the acetylsalicyclic acid (ASA),

Weight = Concentration Volume

2.004mg 0.1L
Weight = ( ) (250cm3 )( )
L 100cm3

Weight = 0.5mg

The calculation is repeated with volume of 50 cm3.

Percentage Difference

Theoretical value = 500 mg = 0.5 g

|Theoretical valueExperimental value|

= 100%
Theoretical value

|5mg5.035mg|
= 100%
5mg

= 0.7%
Discussion

The purpose of this lab was to determine the actual amount of acetylsalicylic acid in the actual
aspirin tablets. Aspirin, is a widely known drug for its analgesic, antipyretic, and anti-inflammatory
properties, is a compound derived from two acids namely acetic acid and salicylic acid. To analyse the
composition of an aspirin sample or the amount of acetylsalicylic acid, the hydrolysis of the sample
by alkali into neutrality is required. By using the Double Beam Ultraviolet Absorption
Spectrophotometry, we were able to measure the absorbance of various dilutions to find the
relationship between the absorbance and the concentration of acetylsalicylic acid in the solution.

First, a stock solution was prepared by weighing 0.1g of pure salicylic acid and quantitatively
transfer to a 100cm3 volumetric flask. The substance was then mixed and diluted with 0.1 mol/L NaOH
solution to form the stock solution. The series of standard solution were then prepared by transferring
the stock solution into a 100cm3 volumetric flask. The concentration of the series standard solution
were 10mg/L, 20mg/L, 30mg/L and 40mg/L respectively.

The graph peaks are labelled and they shows the absorbance of each standard solution. The
graph result is chosen after the 250nm wavelength because the graph form before 250nm is considered
as noise due to the presence of impurities. From the graph, we find that it is a linear relationship and
we are able to calculate the amount of acetylsalicylic acid in the tablets after determining the
absorbance of solution. The amount of acetylsalicylic acid we found in the table was 0.035g more than
the amount stated 0.5g. We found the value is 0.7% more than the stated amount as shown in the
calculation part.

There are some precautions to be taken in order to get a more precise result. The cuvette that
used to contain the standard solutions needs to be wiped cleanly after each test. During the experiment,
we carefully discarded the cuvette and cleaned it every time before we filled it with another standard
solution with different concentration to prevent changes in concentration of the solution. Furthermore,
the way when we inserted the cuvette into the Varian Cary 100 UV-Vis double-beam scanning
spectrophotometer must be taken precisely. The clear and smooth surface of the cuvette must face the
light source and allows the light to pass through. While conducting the experiment, we accidentally
inserted the cuvette with rough surface facing the light source. Therefore, a mistake of curve, attached
in the appendix, which is very different with the other curves, was recorded in our result. Besides, the
original stock acetylsalicylic solution might have some impurities that lead to a secondary reaction that
produced particles that absorb the UV light and affected the experimental result.
 Besides the method of Double Beam Ultraviolet Absorption Spectrophotometry, the
experiment can be conducted by using back titration method, diazotization by using 2,4-
dichloroaniline and HPLC method.

Lab coat and gloves are worn for safety purpose.

Questions

Q1.

In this experiment, salicylic acid stock solution prepared by using 0.1 mol/dm3 sodium
hydroxide (NaOH) because NaOH can hydrolyse the salicylic acid into sodium salicylate. This
reaction assures that the aspirin is 100% hydrolysed into salicylic acid. Then, the salicylic acid will be
diluted by NaOH and reach a certain concentration.

After stock solution has been done, it will obey Beers law and its concentration can be
calculated. The reaction of salicylic acid hydrolysed by NaOH is as below:

C7 H6 O3 + 2NaOH C7 H4 O3 Na2 + 2H2 O

If the salicylic acid is not 100% hydrolysed, it cannot detect the accurate wavelength of salicylic
acid in the experiment because the other compound will affect the wavelength of the solution.

Q2.

From this experiment, the wavelength we obtained is 296nm and 297nm. In reality, the
wavelength of salicylic acid approaches 297 nm so this result can be accepted.

The absorbance of the abfive standard solutions have different values which are 0.278, 0.553,
0.831, 1.064 and 1.233 due to the different concentration of stock solution.

By using Beers Law, the absorbance (A) is equal to molar absorptivity () of solution times
the path length (b) and times the concentration of the solutions(c). In these cases, the path length and
molar absorptivity are constant because of same wavelength which is around 296nm and 297nm.
Therefore, we can generally conclude that the absorbance is proportional to the concentration of
solution.

The absorbance of the solution with ratio of 1:10 is the same with standard solution 2. Thus,
we can confirm that the concentration of the unknown solution is the same with standard solution 2
which is 20mol/L.

Conclusion

As a conclusion, the concentration of salicylic acid in the unknown sample solution is 20

mg/dm3, and the mass of aspirin in the commercial tablet was calculated to be 5.035mg, which has
0.7% difference from the accepted value of 5mg. These errors were caused by real limitation of Beers
Law rather than by human or equipment errors. Hence, the Double Beam Ultraviolet Absorption
Spectrophotometry is still a useful instrument in analytical chemistry.
References

A. M. Helmenstine. (2017). Dilutions from Stock Solutions. Retrieved, 15 June 2017, from
http://chemistry.about.com/od/chemistryquickreview/a/dilutionmath.htm

D. A. Skoog, D. M. West, F.J. Holler, S.R. Crouch. (2014). Fundamentals of Analytical Chemistry,
California: Brooks/Cole- Thomson Learning.,

J. Clark. (2006). A Double Beam Uv-visible Absorption Spectrometer. Retrieved, 15 June 2017, from
http://www.chemguide.co.uk/analysis/uvvisible/spectrometer.html

T. G. Chasteen. (2009). A Double Beam Spectrometer. Retrieved, 16 June 2017, from

http://www.shsu.edu/chm_tgc/primers/spect.html

W. Reusch. (2013). Visible and Ultraviolet Spectroscopy. Retrieved, 16 June 2017, from
http://www2.chemistry.msu.edu/faculty/reusch/VirtTxtJml/Spectrpy/UV-Vis/spectrum.htm