Purification of Acetaminophen by

Recrystallization Method
EXPERIMENT 2
OBJECTIVES

• This experiment will guide students to understand and

learn the techniques and skills in the separation and
purification of acetaminophen using recrystallization
method.
THEORY

• Acetaminophen which is also commonly known as

paracetamol is an amide, a compound that is a derivative
of ammonia that has been reacted with an acidic
substance such as acetic acid.
• Acetaminophen is used to treat many conditions such as
headache, muscle aches, arthritis, backache, toothaches,
colds, and fever.
THEORY

• Recrystallization is a laboratory technique used to purify

solids based on their different solubility. A small amount
of solvent is added to a flask containing an impure solid.
The contents of the flask are heated until the solid
dissolves and then the solution is cooled. A purer solid
precipitate, leaving impurities dissolved in the solvent.
Vacuum filtration is used to isolate the crystals.
THEORY

• The purity of acetaminophen can be determined

qualitatively through its melting point. The presence of
impurities lowers its melting point. If colored impurities are
present, they can be removed by the use of small amount
of decolorizing agent such as carbon.
MATERIALS

• Acetaminophen
• Ethanol
• Ferric chloride TS (1% solution)
• Watch glass
• Beaker (100mL)
• Erlenmeyer flask (250mL)
• Hot plate
• Top Loading balance
MATERIALS

• Stirring rod
• Graduated cyclinder (50mL)
• Water bath
• Spatula
• Test tubes
• Ice bath
PROCEDURES

A. Drug synthesis: Separation and Purification of Acetaminophen

B. Identification Test
Drug synthesis: Separation and Purification of
Acetaminophen
→ Place Allow the solution
Acetaminophen to cool. When Wash with 10 mL
(1.5 g) in a 100 crystals begin to of cold water.
mL beaker. appear, place the
beaker in an ice
bath for 20
minutes. Transfer the filter
Add 20 mL of
If no crystals paper and
distilled water,
appear, scratch crystals to a
heat until the
the inside walls of watch glass and
crystals dissolve.
the beaker. let dry.

If the solution Determine the

boils and crystals Collect the mass of the
remain, add 10 crystals using purified
mL of ethanol. filtration. acetaminophen.
Identification Test

1. Solubility test
– Hot water
– Cold water
– Ethanol

2. FeCl3 Test
• Dissolve 0.1 g of the test substance in 10 mL of water and add 1
drop of ferric chloride TS.
Purification of Acetaminophen by
Recrystallization Method
RESULTS
Solubility Test
Solubility Test
Solubility Test
Solubility Test
FeCl3 Test
FeCl3 Test

Intense violet-blue
color was produced.
Purification of Acetaminophen by
Recrystallization Method
POST LAB DISCUSSION
Questions

1. How do you choose the appropriate solvent in recrystallization

method to isolate and purify certain solid drug preparations?
2. Give the chemical name of acetaminophen and its chemical
structure?
3. What are the usual side effects of acetaminophen?
4. What are the precautions in using acetaminophen?
Recrystallization

• Most common method of purifying solid organic compounds.

– A slow, selective formation of the crystal framework resulting in
a pure compound.
– Relies on the property that for most compounds,as the
temperature of a solvent increases, the solubility of the
compound in that solvent also increases. → Principle of
Recrystallization
Recrystallization Vs. Precipitation

Precipitation
A rapid formation of a solid from a solution that usually
produces an amorphous solid containing many trapped impurities
within the solid's crystal framework.
Steps

1. Find a suitable solvent for the recrystallization;

2. Dissolve the impure solid in a minimum volume of hot solvent;
3. Remove any insoluble impurities by filtration;
4. Slowly cool the hot solution to crystallize thedesired compound
from the solution;
5. Filter the solution to isolate the purified solid compound.
Finding a suitable solvent

• The compound should be very soluble at the boiling point of the

solvent and only sparingly soluble in the solvent at room
temperature.

• The unwanted impurities should be either verysoluble in the

solvent at room temperature orinsoluble in the hot solvent.
Finding a suitable solvent

• The solvent should not react with thecompound being purified.

• The solvent should be volatile enough to be easily removed from

the solvent after the compound has crystallized.
Dissolving the Solid

• Once a suitable solvent is selected, place the impure solid in an

Erlenmeyer flask and add a small volume of hot solvent to the
flask.

• Keep the solution in the Erlenmeyer flask warm on a hot plate or

in a water bath, and add small volumes of hot solvent to the flask
untilall of the solid just dissolves.
Using Decolorizing Carbon

• Decolorizing or activated carbon is used to remove the colored

impurities from the sample.

• Decolorizing carbon is very finely divided carbon that provides

high surface area to adsorb the colored impurities.

• Very little decolorizing carbon is needed to remove the colored

impurities from a solution.
Crystallizing the Solid

• Cover the flask containing the hot filtrate with a watch glass and
set it aside undisturbed to cool slowly to room temperature.

• As the solution cools, the solubility of the dissolved compound will

decrease and the solid will begin to crystallize from the solution.
Crystallizing the Solid

• It may be placed in an ice bath to increase the yield of solid.

• Do not rapidly cool the hot solution by placing the flask in an ice
bath before it has cooled to room temperature.
– This will result in a rapid precipitation of the solid in animpure form
because of trapped impurities.
If No Crystals Formed

• Scratching the inner wall of the Erlenmeyer flask with a glass

stirring rod - to release very small particles of glass which act as
nuclei for crystal growth.

• Another method of inducing crystallization is to add a small crystal

of the desired compound, called a seed crystal, to the solution.

• Seed crystal - acts as a template on which the dissolved solid will

begin crystallizing.
If No Crystals Formed

• If neither of these two techniques results in crystallization, the

compound was probably dissolved in too much hot solvent.

• Reheat the solution to boiling, boil off or distill some of the solvent,
and then allow the solution to cool to room temperature again to
effectcrystallization.
Isolating the Pure Solid

• Once the compound has completely precipitated from the solution,

it is separated from the remaining solution by filtration.

• Vacuum or Suction filtration using a Büchner funnel.

ACETAMINOPHEN

Formula: C8H9NO2
Other names: N-acetyl-para-aminophenol (APAP)
ACETAMINOPHEN

• Acetaminophen normally undergoes glucuronidation and

sulfation to the corresponding conjugates, which together
make up 95% of the total excreted metabolites.
• The alternative P450-dependent GSH conjugation
pathway accounts for the remaining 5%. When
acetaminophen intake far exceeds therapeutic doses, the
glucuronidation and sulfation pathways are saturated, and
the P450-dependent pathway becomes increasingly
important.
ACETAMINOPHEN

• Little or no hepatotoxicity results as long as hepatic GSH

is available for conjugation. However, with time, hepatic
GSH is depleted faster than it can be regenerated, and a
reactive, toxic metabolite accumulates.
SIDE EFFECTS
• Diarrhea
• Increased sweating
• Loss of appetite
• Nausea or vomiting
• Stomach cramps or pain
• Swelling, pain, or tenderness in the upper abdomen or stomach
area
PRECAUTIONS

• Although it is a relatively safe pain medication, several

precautions should be recognized, including not
exceeding the recommended maximum dosage of 4 g/d.
• A lower daily dose of less than 2 g/d is required in
patients who are chronic alcoholics or have renal
complications.
Sources

• Beale, J. M., Jr., & Block, J. H. (2011). Wilson and Gisvold’s Organic
Medicinal and Pharmaceutical Chemistry. China: Lippincott Williams &
Wilkins.
• Katzung, B., & Trevor, A. (2015). Basic & Clinical Pharmacology. USA:
McGraw-Hill Education.
• https://www.drugs.com/sfx/paracetamol-side-effects.html
Synthesis of Salicylic Acid

EXPERIMENT 3
OBJECTIVES

• To be able to prepare salicylic acid from the reaction of

wintergreen oil and sodium hydroxide.
THEORY

• Salicylic acid is a white crystalline compound, which can

be isolated from the bark of birch trees. The pure acid
possesses several useful medicinal properties.
• Unfortunately, pure salicylic acid and even the
concoctions derived from willow, make for an extremely
unpleasant medicine.
THEORY

• The salicylic acid molecule contains two acidic functional

groups, the phenol group and the carboxylic acid group.
• These groups make salicylic acid an irritating substance
that burns the sensitive linings of the mouth, throat,
esophagus, and stomach.
THEORY

• Derivatives of salicylic acid are considered among the

very important compounds in pharmaceutical industry.
Among them are methyl salicylate (oil of wintergreen) and
acetylsalicylic acid (aspirin).
• Although salicylic acid was used historically as an
analgesic (pain reliever), today it is commonly used in
ointments and plasters for the removal of warts from the
skin.
MATERIALS

• Oil of wintergreen (10mL)

• Sodium hydroxide (25%)
• HCl (conc.)
• Bromine water
• Ferric chloride solution (9% solution)
• Methanol
• Sulfuric Acid
• Blue litmus paper/ pH meter
MATERIALS

• Reflux condenser
• Erlenmeyer flask
• Test tubes
• Ice bath
• Watch glass
• Graduated cyclinder
• Spatula
• Iron stand, iron clamp
• Hot plate
PROCEDURES

A. Preparation of Salicylic acid

B. Isolation of Salicylic acid
C. Purification of Salicylic acid

Identification Test
a) Physical Test
b) Chemical test
A. Preparation of Salicylic acid

→ Mix 2.5 mL of oil The Erlenmeyer

of wintergreen flask is then
(methyl salicylate) connected to a Add hydrochloric
with 50 mL of reflux heating set- acid (drop by
Swirl the mixture
Sodium up and allowed to drop) until sample
to dissolve the
Hydroxide (25% boil for twenty pH is adjusted to
precipitate.
conc.) into an minutes until the 2. Heavy white
Erlenmeyer cloudy white color precipitate will
flask. The is dissolved into a occur.
sample will form a clear liquid and
cloudy white color. then allow it to
cool.
B. Isolation of Salicylic acid

Collect the Rinse the solid

Cool the mixture precipitated crude product well with
in an ice-water product by ice-cold water.
bath. filtration.
C. Purification of Salicylic acid

→
Allow the solution After the
Dissolve the Then heat the to cool in an ice recrystallized
product (salicylic solution if needed bath until product has dried
acid) by adding to thoroughly complete completely,
25-50 mL distilled dissolve the precipitation of measure its mass.
water in an product. the salicylic acid,
Erlenmeyer flask. then collect the
precipitate by
filtration.
Identification Test

A. Physical Test
Color:
Odor:
Appearance:
Identification Test

B. Chemical Test

1. Reaction with Bromine water

Add a small amount of salicylic acid to 2 mL bromine water.
Shake the mixture and observe for the discharge of color.
2. Reaction with Ferric chloride solution
Mix 5 mL volume of salicylic acid solution and 5 mL of ferric
chloride solution. Observe for the formation of purple solution.
Identification Test

B. Chemical Test

3. Reaction with Methyl alcohol

In a watch glass, mix together a pinch of salicylic acid, 5 drops
of methanol and 5 drops of concentrated sulfuric acid (under
fume hood). Determine the odor of the product formed.
Synthesis: Reflux Process

 Is the process of boiling reactants while

continually cooling the vapor returning it
back to the flask as a liquid.

 Principle: Condensation

 Cooling water is circulated to condense

escaping vapors.

 Boiling stones keep the boiling solution

from bumping.
Synthesis of Salicylic Acid: Chemical Reaction

2-hydroxybenzoic acid

Molecular fornula: C7H6O3

Synthesis of Salicylic Acid: Reactants
Methyl salicylate – ester
 liquid at room temp.
 volatile at varying degrees
 pleasing to nose – fruity odor
 Oil of wintergreen contains 96-98% methyl salicylate

Sodium hydroxide (NaOH) – base

Purification of Acetaminophen by
Recrystallization Method
RESULTS
Identification tests

• Color – Pink (White)

• Appearance – crystalline powder
• Odor – odorless to slight phenolic odor/characteristic odor
• Solubility – soluble in alcohol, ether, boiling water
• sparingly soluble in chloroform
Reaction with Bromine water (Bromination)
Reaction with Bromine water (Bromination)
• Formation of white precipitate – smells of antiseptic
Reaction with FeCl3 solution
Reaction with FeCl3 solution
• Test for free phenyl hydroxyl (phenol)
Reaction with Methyl alcohol
Reaction with Methyl alcohol
• Formation of an ester - sweet/fruity smell

Alcohol + Carboxylic Acid Ester + Water

R'OH + RCOOHRCOOR' + H2O
Purification of Acetaminophen by
Recrystallization Method
POST LAB DISCUSSION
Questions

1. What are the medicinal uses of salicylic acid?

2. What is the pharmacological classification of salicylic acid?
3. What are the groups of compounds that give a violet color to
ferric chloride?
4. What are the usual products that contain salicylic acid?
Salicylic Acid
 White crytalline compound from the bark and leaves of
Willow tree (Salix sp.) – birch trees

 Salicin – active extract of the bark

 Irritating substance that burns sensitive linings of the mouth,
throat, esophagus and stomach
 acidic functional groups – PhOH and COOH

 Important derivatives: ASA and methyl salicylate

 Commonly used in ointments and plasters to remove warts

Pharmacological Classification
 Salicylic acid is a keratolytic agent.

 It belongs to the same class of drugs as aspirin

(salicylates).

 It works by increasing the amount of moisture in the skin

and dissolving the substance that causes the skin cells to
stick together. This makes it easier to shed the skin cells.
 Medicinal Uses of Salicylic Acid

 It has bacteriostatic, fungicidal, and keratolytic actions

 Topical salicylic acid is also used to treat skin conditions that involve
scaling or overgrowth of skin cells:
 psoriasis (red, scaly patches on some areas of the body)

 ichthyoses (inborn conditions - skin dryness and scaling),

 dandruff, corns, calluses, and warts on the hands or feet

Medicinal Uses of Salicylic Acid

Topical salicylic acid treats acne by reducing swelling and

redness and unplugging blocked skin pores to allow
pimples to shrink.

It treats other skin conditions by softening and loosening

dry, scaly, or thickened skin so that it falls off or can be
removed easily.
Precaution with the use of Salicylic Acid
 Topical salicylic acid should not be used to treat genital

warts, warts on the face, warts with hair growing from

them, warts in the nose or mouth, moles, or birthmarks.

 Do not apply topical salicylic acid to skin that is broken,

red, swollen, irritated, or infected.

Products containing Salicylic Acid
Products containing Salicylic Acid
Products containing Salicylic Acid
Products containing Salicylic Acid