UNIVERSITY OF RWANDA

COLLEGE OF MEDICINE AND HEALTH SCIENCE.

SCHOOL OF MEDICINE AND PHARMACY
DEPARTMENT OF CLINICAL PHARMACY AND PHARMACY
PRACTICES.
BACHELOR WITH HONOUR IN PHARMACY

LECTURER: Dr Théoneste UMUMARARUNGU

LEVEL: I

Date: 5TH AND 6TH MARCH, 2024 on Tuesday and Wednesday

Location : HUYE CAMPUS AT BIO-COMPLEX LAB

Time: On Tuesday from 8:30 am-17:OO pm and on Wednesday from 9:00

am-12:00 pm

MODULE: FUNDAMENTAL CHEMISTRY

GROUP 7: MEMBERS
NAMES REGISTRATION NUMBERS
LIZA GAKIRE Lucretia 223000057
NIYOMURINZI Van Roben 223000646
NIYONZIMA Bonaventure 223001011
NKURUNZIZA Vital 223001957
NSENGIMANA Flugence 223001340
NTAKIRUTIMANA Jean Pierre 223002127
RIBERAKURORA Andre 223000511
RUHIRWA Johnson 223001606
RUKUNDO Jean Claude 223001696
RWIBUTSO Prince 223000975
 LABORATORY REPORT ON THE SYNTHESIS OF
ACETAMINOPHEN.

PART I: INTRODUCTION

1. LITERATURE REVIEW(BACKGROUND OF ACETAMINOPHEN)

Acetaminophen is also known as paracetamol and it is mostly used as an antipyretic and

analgesic1. Acetaminophen is preferred alternative to aspirin (acetylasalicylic acid) because
have the same effect. Acetaminophen is usually used as pain reliver and has got the brand name
of Tylenol2.

Acetaminophen is crucial in experiment during studying the effectiveness of Infrared and

Raman spectroscopy in both qualitative and quantitative determinations3. In laboratories
Acetaminophen is easily to synthesize and stable as it is synthesized from reacting para-
aminophenol and acetic anhydride in presence of sulphuric acid to produce Acetaminophen and
acetic acid4.

There are different schools that recently have conducted this experiment and one them is
Arizona State University in United state where they did this same experiment in Lab number of
PSH 334 from 10:35a.m-1.25p.m by Augushi Parikh with Lab partner Doree Dispo on
21stoctober 2021 and the crude acetaminophen they obtained had melting point of 166.2oc-
168oc and had percentage purity of 44.3% but firstly Acetaminophen was synthesized by
Joseph Von Mering in 1893 and was found that the melting point of pure Acetaminophen is
between 169-170.5oc5.

2. AIMS OF THE EXPERIMENT

The aim of our experiment was to synthesize crystallized Acetaminophen by reacting p-
aminophenol with Acetic anhydride and carrying out its quality Analysis using Thin layer
chromatography.

PART II: Methods:

1. LIST OF REAGENTS AND PRODUCTS IN PREPARATION OF

ACETAMINOPHEN.

Reagents:

 para-aminophenol

 Acetic anhydride

 Distilled water
Products:

 Acetaminophen
 Acetic acid

Reagents used for Thin Layer Chromatography(TLC)

 ⮚ Hexane
⮚ Ethyl acetate
⮚ Methanol
The volume ratio is 16:12:2 respectively.
Other reagents used are Standard Acetaminophen tablets and Ethanol
2. LIST OF EQUIPMENTS

⮚ 100ml round-bottomed flask

⮚ aluminium foil
⮚ Heating mantle
⮚ Clamp and stand
⮚ Spatula
⮚ Vacuum pump
⮚ Filter paper
⮚ Glass rod
⮚ TLC Plate
⮚ TLC Tank
⮚ TLC Spotter
⮚ Hirsch funnel
⮚ Electric Oven
⮚ Pencil
⮚ Buchner flask
⮚ Electronic beam balance
⮚ Ruler
⮚ Air condenser
⮚ Goggles and gloves
⮚ U.V Lamp
⮚ White tissue
⮚ Measuring cylinder
⮚ 5ml Graduated pipette

⮚ Digital Melting point Apparatus

⮚ Mortar and pestle

3.HAZARDS ASSOCIATED WITH THE REAGENTS

During our experiment there was various chemical hazards associated with the reagents we
have used where everyone should take care as to avoid those hazards as to keep him or her safe.
Chemical hazards can be irritation,sensitization, carcinogenicity and also can be physical
hazards such as flammability,corrosion and explosibility6.

1. Chemicals hazards of reagents used:

A. para-aminophenol:

para-aminophenol has been found to have different effects to human health as It has acute
health effects, chronical health effects and long term effects which can be developed by
someone when contact, inhaled and swallowed7.

When para-aminophenol gets cantact with the skin can cause skin irritation and also lead to
irritation of the eye8. The high level of para-aminophenol in the blood this lead to reduction
in capacity of the red blood cells to transport oxygen gas in the blood which further cause
headache, fatigue,dizziness, blue coloring of the skin and eyes9.

In addition, para-aminophenol has chronical health effects as it may cause mutation in

genetic make up which may lead development of cancer10 and also it may cause skin
allergy which further cause itching and skin rash, para-aminophenol lead to kidney failure
when become accumulated in the body11.

B. Acetic anhydride:

Acetic anhydride has great health hazards which can be seen after immediately exposure to it.
Some health hazards of acetic anhydride can cause irritation of the skin,eyes and redness of
skin and eyes which can further cause loss of vision12.

It has been found that Acetic anhydride is flammable above 39oc

Which lead to fire in laboratories13.

2. Chemical hazards of products produced:

A. Acetaminophen:

Acetaminophen as it is mostly used as an antipyretic and analgesic but has also serious health
hazards because when the skin and eyes get contact with acetaminophen cause irritation and
development of skin allergy14.
When it is inhaled has been found that acetaminophen cause respiratory irritation which further
can lead to lung damage15.

Acetaminophen has effect to environment as it is toxic to aquatic animals where it is long term
effect hazard to environment where aquatic animals especially fish die and cause loss of
ecosystem16.

B. Acetic acid:

Acetic acid has health hazards when is inhaled or when you get contact with it, because it is
corrosive to eyes,skin and respiratory tract as acetic acid when it is inhaled can cause lung
oedema and prolonged contact with the skin cause dermatitis17.
3. Hazards associated with TLC Reagents
A. Methanol:
Methanol has great health hazards such as can cause skin irritation and prolonged or repeated
contact with it lead to redness of the skin and when it is ingested lead to nausea and vomiting18.
Exposure to high concentration of methanol lead to headache, dizziness and fatigue and also it
is flammable19.
B. n-Hexane:
When you exposure to high concentration of hexane can lead to headache, feel dizzy, fatigue
and hexane damage the liver20. In addition hexane is flammable as it is dangerous on fire21.
C. Ethyl acetate:
Ethyl acetate also has serious health hazards as it cause drying and cracking of the skin in
contact with it and cause liver and kidney failure as it affect kidney and liver functions22.

4. PROCEDURES OF THE EXPERIMENT

Experiment was started by taking a dry round-bottomed flask of 100 ml and 1.0006g of p-
aminophenol( a brown powder) was measured by using an electronic beam balance and placed
in that round-bottomed flask by using aluminium foil as to avoid spread of powder to the
surrounding. After that ,3.0 ml of distilled water was measured by using a graduated pipette
and poured into that round-bottomed flask and we make sure that all powder of para-
aminophenol on the aluminium foil has cleaned into the flask as the way to minimize the errors
and then 1.1ml of acetic anhydride was measured by using graduated pipette and added to that
round-bottomed flask.

After, the round-bottomed flask was connected to an air condenser fixed on clamp and stand
and it’s inlet tube was connected to the tap water as outlet tube was left in the basin. Inlet tube
allows cold water to get in the air condenser and outlet tube allows condensed vapors to leave
the air condenser in form of liquid to the basin and then round-bottomed flask was buried
nearly at the bottom of sand bath of the heating mantle so that it can absorb heat as fast as
possible and the charger of heating mantle was plugged to electricity and switched on.

As the solution was heated, the powder of p-aminophenol started slowly to be dissolved. After
a few minutes, the mixture in round-bottomed flask started to boil, forming bubbles and water
vapors was observed inside, indicating that our solution reached the boiling point.
After 10 minutes, heating the solution was stopped as it’s components had dissolved and
heating mantle was left behind to allow the solution to cool down in 13 minutes.

After cooling down, The air condenser was detached from the round-bottomed flask then by
using a glass rod we scratched inside of the round-bottomed flask as to initiate crystallization in
order to get crude acetaminophen, and then our flask was placed in round-bottomed flask
holder for further cooling and crystallization of the solution.

After two hours , we cut a filter paper that has the same circumference as the open of the
Hirsch funnel and then the mass of that filter paper was found to be 0.5027g by using
electronic beam balance.Then process of filtering the solution started by putting that filter
paper on the Hirsch funnel placed on the Buchner flask which was connected to a vacuum
pump and ethanol was poured on the filter paper as to make it damp and fit into the Hirsch
funnel then after crude acetaminophen mixture from round-bottomed flask was poured on the
filter paper inside the funnel where the power switched on to initiate filtration. After crude
acetaminophen remained on filter paper which was dark brown solid and taken to the
electronic oven for making it dry completely for fifteen hours until next day.

After that, TLC Analysis started before the end of our first day of experiment to find whether
the crude acetaminophen obtained from the experiment has the same content as standard
acetaminophen and three solutions for TLC plate spotting were prepared . Firstly, standard
acetaminophen tablets was ground into powder by pestle and mortar. Secondly, three beakers
were rinsed by ethanol to avoid contamination of solutions. After that, Small amount of crude
acetaminophen from filter paper was poured in one beaker followed by addition of small
amount ethanol then powder of standard acetaminophen was poured in second beaker
followed by adding small amount of ethanol finally small amount of para-aminophenol was
poured in third beaker followed by also addition of small amount of ethanol and all beakers
were labeled by the name of solution that they contain.

After that we prepared a mobile phase for TLC analysis where 16 ml of Hexane, 12 ml of
Ethyl acetate, and 2 ml of Methanolwere measured by using a graduated pipette with pipette
filler and poured into TLC Tank (a transparent cuboidal glass container).

After, we waited for fifteen minutes so that the equilibrium of mobile phase reached between
three solutions used. By the time we were waiting for the equilibrium to be reached we were
spotting the TLC Plate( a rectangular plate that had two different colored sides, the front side
had light white, and the back one had silver-white color)23. We had drawn a horizontal line that
has three spaces along in the distance of 1cm of line where we spotted the solutions that we
used in TLC analysis and that horizontal line we had drawn it above 2.0 cm from the bottom of
the TLC plate using a pencil and a ruler .

After that, we took TLC spotter( micro-cylinder having one open end and other closed end)24
and was rinsed by ethanol, dried by using white tissue and used to spot three solutions from
beakers by spotting each of the three solutions six times to make sure that spots look clear as
good. The first spot was spotted as a solution of p-aminophenol, the second spot was spotted as
a solution of standard acetaminophen and the last was spotted as a solution of crude
acetaminophen we got from the experiment but we washed and rinsed TLC spotter by using
ethanol for every solution we were going to spot to avoid contamination and also dried on
white tissue.
After a short period of time as we had waited for the spot to become dried and after TLC Plate
was inserted in the TLC Chamber that contains the prepared mobile phase and left it for fifteen
minutes.

After that 15 minutes, TLC Plate was removed and directly the line was drawn where the
solvent had reached by using a pencil. After that, TLC Plate was taken and carried to be
observed under the Ultra-Violet Lamp to see clearly the spots and the distance moved by them
and a picture of TLC Plate was taken by using a phone so that we can see it clear when we
were calculating the retention factor.
 After fifteen hours on next day, The filter paper was removed from the oven and we weighed
the mass of it with dried acetaminophen by using an electronic balance and we obtained a mass
of 1.3738g.
PART III: Results and Discussion

A. RESULTS WE OBTAINED FROM THE EXPERIMENT.

Figure1: Reaction between p-aminophenol and acetic anhydride

Table1. Shows the data we had used during experiment.

Chemicals and items Mass/ gram Volume/ ml

p-aminophenol 1.0006 -
Acetic anhydride - 1.1000
Water - 3.0000
Filter paper 0.5027 -
Filter paper and product 1.3738 -
Crude acetaminophen 0.8711 _

Molar Mass of Para-Aminophenol (C6H7NO): = (6 x Atomic Mass of Carbon) + (7 x Atomic Mass of

Hydrogen) + Atomic Mass of Nitrogen + Atomic Mass of Oxygen = (6 x 12.01) + (7 x 1.008) + 14.01 +
16.00 = 72.06 + 7.056 + 14.01 + 16.00 = 109.126g/mol
Therefore, the molar mass of para-aminophenol is 109.126g/mol

We measured the mass of p-aminophenol of 1.0006g and by considering the percentage purity
of p-aminophenol of 98%, we found the actual mass of it
98
as:������ ���� = 100x1.0006=0.9805g

0.9806
Number of moles of para-aminophenol= 109.126=8.98x10-3moles
According to the chemical equation on figure1 we can calculate the number of moles
of acetaminophen produced by considering the molar ratio 0f 1:1 from the equation.
Then the number of moles of acetaminophen is 8.98x10-3moles and now we can now
 calculate the mass of acetaminophen produced.
C8H9NO2 = (8 × molar mass of carbon) + (9 × molar mass of hydrogen) + (1 × molar
mass of nitrogen) + (2 × molar mass of oxygen).
C8H9NO2 = (8 × 12.01 g/mol) + (9 × 1.01 g/mol) + (1 × 14.01 g/mol) + (2 × 16.00 g/mol)
C8H9NO2 ≈ 151.16 g/mol + 9.09 g/mol + 14.01 g/mol + 32.00 g/mol ≈ 206.26 g/mol.
Therefore, the molar mass of acetaminophen (C8H9NO2) is approximately 206.26 g/mol. Now
mass of acetaminophen is 8.98x10-3molesx206.26g/mol=1.85g
And this mass was theoretical yield we had expected to get but the actual mass of
acetaminophen we obtained from experiment was 0.8711g.

We found out also which reactant was limiting reagent as we had the density of acetic
anhydride,percentage by mass of it which was 98%MW on the bottle and its volume used .
The density of acetic anhydride was 1.08g/cm3 on the bottle and volume we used was 1.1cm3
����
that means there is 98g of acetic anhydride in 100g of solution. ������� = ������ , volume
���
of acetic anhydride in the stock is = �.���/��=91ml
Molar mass of acetic anhydride is (4 * 12.01 g/mol) + (8 * 1.01 g/mol) + (2 * 16.00 g/mol) =
102.09 g/mol
����
Number of moles of acetic anhydride in 91ml= �� ,
98�
Now, it’s moles is = 102.09�/���=9.59x10-1moles in 91ml. Therefore the number of moles of
�.�����−������
acetic anhydride in 1.1ml= ��
x1.1=1.16x10-2moles and according to the chemical
equation the molar ratio between acetic anhydride and acetaminophen is 1:1 therefore also the
number of moles of acetaminophen is 1.16x10-2moles and now the mass of acetaminophen
1.16x10-2molesx206.26g/mol=2.39g and this comfirmed that acetic anhydride is excess reagent
of the reaction.

Therefore, we can calculate the percentage yield of acetaminophen produced by using

theoretical mass we obtained from p-aminophenol as it is limiting reagent by using this
formular where percentage yield=(actual yield/theoretical yield)×100
0.8711
��������� ����� =
1.85
x100=47.1%.

We had measured the melting point of crude acetaminophen by using Digital Melting Point
apparatus by setting temperature of 100oc and Rise temperature of 10oc/min and we found that
crude acetaminophen melting point between 170.1oc-175.5oc.

RESULTS FROM TLC PLATE

 Distance moved by solution from mobile phase

Spot of impurities from p-aminophenol

Spot of impurities from crude acetaminophen

Distance for mobile phase

Figure2: TLC plate image from the TLC chamber under U.V lamp

Distance moved by spots:solvent=5.7cm,Product=2.5cm,SA=2.4cm,PA=2.5cm

TLC Plate shows three spots, the first is for p-aminophenol, the second for standard
paracetamol in middle and the last one on right side of TLC plate corresponds to the crude
acetaminophen.
CALCULATION OF RETENTION FACTOR

Retention factor(Rf) is calculated as the distance traveled by the compound divided by the
distance traveled by the solvent front25.
�.�
Retention factor for p-aminophenol=�.�=0.43
�.�
Retention factor for standard acetaminophen=�.�=0.42
�.�
Retention factor for crude acetaminophen=�.�=0.43

DISCUSSION OF THE OBTAINED RESULTS

We had obtained the percentage yield of 47.1% not higher as much as we was expected to get
100% because of different reasons such as the all para-aminophenol had not reacted
completely with acid anhydride, some powder of para-aminophenol had spread to the
surrounding which reduced the mass of it, the equipment we used during experiment might be
contaminated and not well cleaned by ethanol used to rinse them and cause interference of
para-aminophenol with impurities, there might be loss of mass during filtration because not all
acetaminophen has crystallized, mass of acetaminophen we using during TLC analysis might
be the cause and also we did not use sulphuric acid as catalyst where others did the same
experiment used it and by comparing our results with experiment carried out in Arizona State
University in United state they got percentage yield of 44.3% and they had obtained melting
point of 166.2oc-168oc where for us we got melting point of 170.1oc-175.5oc and this shows
that we tried to do better than them as our melting point we got approaches the melting point of
pure Acetaminophen which is between 169-170.5oc5.
By discussing about the results we obtained TLC analysis we had seen that our crude
acetaminophen had moved higher a little bit distance than a standard acetaminophen which
caused by crude acetaminophen is less polar than standard acetaminophen and this helped us
to confirm that our product we obtained from the experiment also contains acetaminophen as
both they shown blue coloured spots on TLC plate and also the retention factors for them are
mostly similar.

Part IV: Conclusion

A. BRIEF SUMMARY OF THE OBTAINED RESULTS

In summary, we had obtained the percentage yield of 47.1% which was not higher as 100% for
different reasons we have mentioned in part of discussion and by comparing to others who did
the same experiment shows that our experiment had become more success than their
experiments like one that conducted in Arizona State University in United state they got
percentage yield of 44.3% and they had obtained melting point of 166.2oc-168oc where for us
we got melting point of 170.1oc-175.5oc and this shows that we tried to do better than them as
our melting point we got approaches the melting point of pure Acetaminophen which is
between 169-170.5oc5.

By focusing on results obtained from TLC analysis showed that our experiment of synthesize
acetaminophen was successful as our product and standard acetaminophen had shown the same
spots of blue coloured on the TLC plate and their retention factors was mostly the same.

B. RECOMMENDATIONS
By looking through whole experiment we conducted of synthesize acetaminophen from para-
aminophenol we may recommend the following as to make easier next experiments that will be
conducted by other classes on the same experiment, such as to buy electric socket where
students will use to connect air condenser to electricity and to make sure that all laboratory
equipment are well prepared before the experiment starts because affected us much to move to
next room to find them which lead to delay of the results.

Part V: References

1) Pasquarella, K; Jardine, K; Hill, K; Jones, E; Elia, R; Gibbs, G; Sonntag, M; Tribe, L.

Synthesis and characterization of acetaminophen: An experimental and theoretical
laboratory for the undergraduate curriculum. J. Chem. Educ. 2021
2. Tribe, L.; Kostka, K. Peer-Developed and Peer-Led Labs in General Chemistry. J. Chem.
Educ. 2007, 84, 1031.
3. Prescott, L. Paracetamol: past, present, and future. American Journal of Therapeutics. 2000
7(2), 143-148
4. New Jersey Department of Health Senior Services: Hazardous Substances Fact Sheet(online
document on aminophenol).2007.Available from
https://www.nj.gov/health/eoh/rtkweb/documents/fs/0078.pdf
5. New Jersey Department of Health Senior Services: Hazardous Substances Fact Sheet(online
document on acid anhydride).2005.Available from
https://www.nj.gov/health/eoh/rtkweb/documents/fs/0005.pdf
6. New Jersey Department of Health Senior Services: Hazardous Substances Fact Sheet(online
document on acetaninophen).2008 pages:1-6.Available from
https://cdn.caymanchem.com/cdn/msds/22629m.pdf
7. New Jersey Department of Health Senior Services: Hazardous Substances Fact Sheet(online
document on acetic acid).2007.revision 2016 1.Available from
https://www.nj.gov/health/eoh/rtkweb/documents/fs/0004.pdf
8. Brune K. The early history of non-opioid analgesics. Acute Pain
1997; 1: 33-40.
9. Drews J. Drug discovery: a historical perspective. Science 2000;
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10. Morse HN. Ueber eine neue darstellungsmethode des acetylamido
phenole [A new method of preparing acetomidophenole]. Ber
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11. Cahn A, Hepp P. Das antifebrin, ein neues fiebermittel [Antifebrin, a
new antipyretic]. Zentralbl Klin Med 1886; 33: 561-4.
12. Drugs discovered through serendipity in the laboratory. In: Sneader
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