A COMPILATION OF SYNTHESES OF SELECTED

ORGANIC COMPOUNDS AND NATURAL PRODUCTS

(A Laboratory Manual for Pharmaceutical Chemistry 2)

Henedina A. Maini, RPh

May 2014

FOREWORD

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 The experiments included in this manual have been selected
from various sources and were chosen in order to aid students in
their understanding of how a pharmaceutically important compound
is produced synthetically from organic and inorganic chemicals
commonly seen in the College of Pharmacy laboratory. The process
of preparation also includes testing for the identity of the products
produced using official identity tests coming from available editions
of the United States Pharmacopoeia and the National Formulary and
determination of physico-chemical constants, such as melting point,
boiling point and solubility characteristics, to name a few.

It is also a means by which students studying Pharmacy would

be introduced to the rudiments of laboratory work, giving them
ample exposure in the implementation of procedures as presented
to them in the different experiments, as well as making them
improve on their skills acquired through years of studies, instilling
in them the value of time, honesty, patience and hard work.

The experiments have been validated in order to minimize

unexpected reactions that might arise from the conduct of the
same.

It is our hope that this simple contribution would instill in the

students the importance of understanding the basics of Organic
Chemistry in relation to the biologically-active compounds
identified.

H. A. Maini, RPh

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 PROPER CONDUCT OF STUDENTS IN THE LABORATORY

It is expected of every student enrolled in Pharmaceutical Chemistry 2

to observe the following:

1. WEARING OF PROPER PPE (PERSONAL PROTECTIVE EQUIPMENT)

The safe way is the right way to do your experiment. It is without

saying that students have to be protected from the hazards of conducting an
experiment might bring, thus, a student is expected to have the following
minimum requirements when they enter the laboratory:

1. Laboratory gown
2. Laboratory shoes
3. Head cap
4. Safety goggles
5. Face mask
6. Gloves

Safety goggles should be worn at all times inside the laboratory.

Personal effects: wear proper clothing (including protective clothing when
handling corrosive, toxic, or flammable materials). Avoid loose sleeves, loose
cuffs, and bracelets. Be careful with long hair. Proper shoes are required (no
sandals).

On top of this, students in a group should have in their lockers the following:
7. Test tube holder
8. Test tube brush
9. Soap solution or detergent
10. Bottle brush
11. Oil cloth
12. Rags
13. Plastic bags
14. Pot holders
15. Various sizes of corks
16. Scissors

Be able to use all safety devices and protective equipment provided for
your use and know their location (eyewash fountain, shower, fire
extinguisher).

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 This minimum requirement would ensure to some extent that a student
would be partly ready in conducting the experiment that is assigned for the
day. Of course, other requirements for the experiment should be requested
from the Instrument Room.

2. DO NOT EAT OR DRINK IN THE LABORATORY (and do not store food in

the refrigerators). Smoking in the laboratory is absolutely forbidden.

3. HORSEPLAY in any form is dangerous and prohibited. Do not run in

laboratory areas.

4. REPORT TO YOUR LABORATORY INSTRUCTOR ALL UNSAFE

CONDITIONS, unsafe acts, and "near misses" that might cause
future accidents. Report any accident or fire, no matter how trivial, to the
laboratory Instructor.

5. HAZARDOUS CHEMICALS:
a. Be especially mindful of fire hazards when you or your lab neighbors
are working with flammable liquids.
b. Hazardous Substances: Know common explosive, toxic, and
carcinogenic materials and use them only with adequate
safeguards.

6. NEVER LEAVE A REACTION OR EXPERIMENT RUNNING

UNATTENDED, unless you have told your lab partners enough about it to
deal with potential hazards while you are away.

7. KEEP HOOD AND BENCH TOPS AREAS CLEAN AND WORKABLE

SPACE MAXIMIZED.

8. ADVANCED READING OF THE EXPERIMENT

Students are expected to have a preliminary reading of the exercises in
order for them to be aware of the conditions and requirements of the
experiment/s scheduled for the day. Plan your work. Follow instructions. If
after reading the instruction and procedure you still do not know how to do
the experiment safely, ask your laboratory instructor. Always be ready for
a quiz on the scheduled experiment.

9. PREPARATION OF THE PRE-LAB REPORT

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 Preliminary data should be obtained for the compound to be
synthesized.

10. REQUISITION OF CHEMICALS AND GLASSWARE

Requisitions for the said experiment scheduled to be performed should
be requested from the Instrument Room THREE (3) DAYS BEFORE THE
SCHEDULED EXPERIMENT by filling up the necessary forms pertaining to
the needs of the experiment. Requisitions should be SIGNED BY THE
LABORATORY INSTRUCTOR as well as the STUDENT REPRESENTATIVE OF THE
GROUP MONITOR identified or assigned for the said experiment.

Test solutions and chemicals needed for the experiment should also be
prepared (if there is a need to).

11. FOCUS AND PRESENCE OF MIND IN THE CONDUCT OF THE

EXPERIMENT/S.
There is no substitute for COMMON SENSE.
Simple rules learned through the study of chemistry, e.g. “Acid to
water NOT water to acid”, in the previous year levels would certainly help the
student in hurdling the chemical reactions that he/she will meet in this
course. Thus, focus and presence of mind during the conduct of the
experiment/s is A MUST.

LABORATORY HAZARDS OVERVIEW

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Chemical Hazards
Every chemical is a poison, depending on the dose taken. We handle
hazardous chemicals in our everyday lives, from the gasoline we use to run
cars to using chlorine bleach to clean the laundry. The keys to safe use of
these and any chemical are: understanding the hazards of each chemical;
knowing and using safety measures to minimize these hazards. However, it
is easy to forget all these and treat hazardous materials casually.
Precautionary measures have to be taken seriously. Simple precautions can
be a lifesaver; use them as you use a harness when driving a car.

Liquid Chemical Hazards

Liquid chemicals present the greatest potential risk for injury:
1. they have to be handled (transported, poured, and mixed) to be
used;
2. there’s a wide variety, each with a different set of hazards and
precautionary
measure required for use.

Chemicals in the laboratory can cause severe burns, tissue, and organ
damage, and can ignite and explode. The greatest health risks posed by
liquid chemicals are physical (fire, explosion), direct contact with skin and
eyes (tissue damage), and inhalation (pulmonary damage or long term
chronic effects).
For detailed hazard information, consult the pertinent MSDS.
Make every effort to understand the chemical processes you use and
respect the chemicals you work with. Knowing the general rules on how to
safely transport, pour, use, and dispose of these chemicals is every
laboratory user's responsibility.

Gas Hazards
Compressed gases pose both chemical and physical hazards. Some of
the gases used are inert; others are toxic, corrosive, flammable, or explosive.
The primary health risks posed by gases are the physical hazards (fire,
explosion) and inhalation (toxins and corrosives). Because of these potential
hazards, as a laboratory user, one must be always aware of the types gases
and the hazards posed in the equipment being used.

Electrical Hazards

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 Electrical shock hazards are present wherever electricity is used.
Although equipment is interlocked to prevent operator exposure, one must
be aware the electrical hazards for the tool you are using. Burns occur
wherever the body completes a circuit connecting the power source with
ground. Although the resistance of dry, unbroken skin to electric current is
relatively high, the amount of current needed to kill a person is small. It is
easy to exceed lethal levels of current, especially if the skin is broken, wet, or
damp with sweat.
Unless it is in your training, never open electrical enclosures or
cabinets on equipment, even when the power is off. If you feel an electrical
"tingle" when you touch a piece of equipment, stop using the tool and
immediately notify a maintenance staff person. Never stick your hands,
fingers or conductive tools inside equipment. Immediately notify the
maintenance staff of any potential electrical hazard that one notice.
Electronic devices that are not allowed to be used in the laboratory
are personal
listening devices (iPods and such) as these may prevent the user from
hearing alarms or lab announcements.
Other Hazards
Ultraviolet Radiation UV exposure is a potential risk.
High power UV lamps as they are mercury-based, they pose a chemical
risk. If a UV lamp should break or explode, do not attempt to clean up;
instead, isolate the immediate area and call staff.
Electromagnetic Radiation may be generated by equipment. Report any
damage to shielding on the equipment or cables.

Liquid Chemicals
Liquid Chemical Hazard Classes
Corrosive:
A corrosive (or "caustic") chemical destroys or permanently damages
living tissue. On contact, corrosives can destroy skin and underlying tissues.
Splashes in the eyes can cause blindness. Inhalation of vapors can destroy
lung tissue.
Corrosives in the laboratory include acids and bases. In case of
localized external exposure, promptly flush the affected area with plenty of
water, for at least 15 minutes. Remove clothing while under the shower and
flush for at least 15 minutes. Exposure of corrosives to the eyes is extremely
serious; flush immediately, either with a spray gun at your wet bench or the
nearest eyewash station. Eyes should be rolled up and down, and side to
side, continuously, to allow clean water to flush behind the eyeball. For any

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exposure to corrosives, one should get help. The victim should be taken to
the emergency center for evaluation and treatment.

Oxidizing agent
An oxidizing agent is a chemical compound that has a pair of electrons
to donate to an electron-accepting, reducing agent. Often, they contain
reactive oxygen. When mixed with compounds that can act as reducing
agents, the result is often a violent reaction, possibly an explosion.
Oxidizing agents should not be stored or mixed with solvents, which
generally make
excellent reducing agents. Oxidizing agents are stored in the chemicals pass-
through. One oxidizing agent is hydrogen peroxide (H2O2). Nitric acid
(HNO3) is an oxidizing agent as well as a corrosive.
In the laboratory, the main principle behind segregation of chemicals is
to keep oxidizing agents away from flammable chemicals (namely, solvents)
and any combustible materials (some chemicals, materials like laboratory
wipes).

Water reactive
Water reactive describes compounds which very quickly generate heat
and/or gas upon mixing with water. These are often concentrated acids or
bases. The primary hazard presented by water-reactive compounds is
incomplete mixing, which can lead to superheating and explosion. Thus,
water-reactive mixtures should never be poured directly into a sink drain.
Aspirating water reactive mixtures at the wet benches is standard
practice; the high dilution factor and rapid mixing dissipates heat and
prevents superheating.

Flammables
Flammables include most solvents, such as acetone, isopropanol, and
methanol. The “flash point” of a flammable is the concentration in air above
which the vapors from a flammable can ignite and explode. The source of
ignition may be heat (such as a hot plate) or a spark (such as from an
electrical tool). Because the vapors can travel over considerable distances,
the source of ignition can be far away from the flammables container itself.
To minimize hazards, always work well within the exhausted area of the
appropriate bench (behind the red line). The air pulled into the exhaust area
will keep the concentration of vapors below the flash point. Where possible,

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minimize the quantities of flammables used. Before working with
flammables, always note the location of the nearest safety shower and
fire extinguisher. Flammables should be stored in the designated
flammables’ cabinet; no flammables may be stored in the laboratory.
Flammables must be kept away from oxidizing agents.

Toxin/Poison
A toxic material is one that has poisonous or harmful effects. There are
formal, quantifiable definitions as to what comprises a toxic material and to
what degree it is toxic. These definitions are based on lethal dosages for
laboratory animals when administered orally or through inhalation.

Non-toxic
A non-toxic material is one that is not likely to result in harmful effects
with normal use. This designation is used sparingly. Pure water is considered
non-toxic.

Specific Hazardous Liquid Chemicals

Solvents
Flammable Solvents
Acetone, isopropanol, and methanol may be found in the solvent wet
benches. These chemicals are all flammable solvents with low flash points.
This means that at sufficiently high vapor concentrations, they can be easily
ignited at room temperature and, therefore, pose significant fire hazard.
Thus, solvents should not be used on or near hot plates or near any
electrical system. Solvents may also ignite or explode when brought into
contact with chemical oxidizing agents (such as many acids) and so should
not be mixed with, nor collected in the same waste container as these
compounds.
Standard solvent waste should be disposed of in the solvent carboy or
collected locally.
These and other solvents must be stored in the designated flammables
cabinet in the service area and may be transported in the laboratory only if
carried in metal carts. Solvents may be used only in designated solvent
hoods.

Chlorinated Solvents
Chlorinated solvents (such as chlorobenzene, trichloroethylene [TCE],
and methylene chloride) may be present in some processes, although these
have been phased-out of general use. Long term, repeated exposure to some

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chlorinated solvents is correlated to cancer and liver and nerve damage.
Because of environmental hazards, chlorinated solvent waste must be
collected in a waste container, separate from other kinds of liquid solvent
waste.

Oxidizing Agents
Peroxides
All peroxides are highly oxidizing materials; energy is released when
they are reacted. Some peroxides are unstable, and can explode. Extreme
care should be used in mixing solutions containing peroxides. Peroxides are
incompatible with all forms of organic solvents and flammable materials.

Nitric Acid
Nitric acid is also water reactive (heating upon addition of water). All
oxidizers should be kept away from solvents, bases, and flammable
materials.

Alkali/Bases
Alkaline compounds, or bases, are the chemical opposite of acids, and
may react violently when mixed with them. They are most commonly used in
the laboratory in lithography and etch. Alkalis are caustic, so protective gear
should always be worn when working with them to prevent contact with skin
and eyes.

Specific Hazardous Gases

Corrosive Gases
Hydrochloric acid (HCl)
HCI gas is extremely corrosive to almost everything, including stainless
steel. It is used for cleaning/etching deposition chambers.

Ammonia (NH3)
NH3 gas is a severely corrosive alkaline vapor with a pungent odor. It is
shipped in the cylinder as a liquid under its own vapor pressure of
approximately 9 atm. NH3 gas is used in oxynitride and nitride film
deposition (plasma and CVD.)

WHAT TO DO IN AN EMERGENCY

TYPE OF EMERGENCY RESPONSE

Fire Alarm Sounds • Leave the building immediately.

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 • Meet at the Evacuation Assembly Point
• DO NOT re-enter the building until cleared.

Health Threatening
Fire, toxic spills, gas leak
• Pull the fire alarm, if alarm is not already sounding.
• Leave the building immediately.
• Meet at the Evacuation Assembly Point
• DO NOT re-enter the building until cleared.

Major Earthquake
• Take cover; wait for shaking to stop.
• Leave building & meet at Evacuation Assembly Point
• DO NOT pull the fire alarm unless there is a health-threatening
hazard.

Life-Threatening Medical Emergency

(if in doubt, treat as an emergency)
• Call out for help.
• Dial the emergency number immediately (DO NOT hang up until told
to do so.)
• DO NOT move victim unless necessary.
• First aid kit/AED/Oxygen Units located near the instrument Room

Non-Health Threatening Emergency (Building and facilities)

• During work hours: call Kaycie Baliza at the Dean’s Office; in the
laboratory, call Ryan Quiming.
• After hours, call ST Gate information desk.

Electrical Power Outage

• Emergency backup lights should come on within 15 seconds. (normal
building
operations will be interrupted until regular power is restored)

Chemical Spill
• Isolate the affected area.
• Leave immediate area and make sure others leave also
• Work hours: page Kaycie Baliza of the Dean’s Office or Ryan Quiming
of the Instrument Room
• Off hours: Call ST Gate Information Desk

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Odor in the Lab
• Leave immediate area and make sure others leave
• Work hours: page staff and call Ryan Quiming of the Instrument
Room

FORMAT OF LABORATORY REPORT

The laboratory report will be a group report. Laboratory reports,

composed of the pre-lab and post lab reports, should be written in short bond
paper to facilitate the submission of the compiled report at the end of the
semester. Writing of laboratory report should follow the following format:

1. PRE-LAB REPORT (30 pts)

EXPERIMENT NO.
TITLE OF THE EXPERIMENT

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GROUP NO. START OF
EXPERIMENT (DATE)
NAMES OF GROUP MEMBERS END OF EXPERIMENT
(DATE)

I. OBJECTIVES: (Minimum of 2, maximum of 5)

II. DIAGRAM OF THE PROCEDURE
III. PRELIMINARY DATA ON THE COMPOUND TO BE PREPARED
A. PHYSICAL APPEARANCE
B. PHYSICO-CHEMICAL PROPERTIES, EX. MELTING POINT (IF SOLID),
BOILING POINT (IF LIQUID)
IV. CHEMICAL REACTION INVOLVED IN THE SYNTHESIS
V. COMPUTATION FOR THE THEORETICAL YIELD
VI. TEST FOR IDENTITY OF THE COMPOUND TO BE PREPARED
(MINIMUM OF TWO TESTS, WITH THEIR CORRESPONDING
CHEMICAL REACTIONS)

2. POST-LAB REPORT (70 PTS.)

VII. DATA/RESULTS OBTAINED FROM THE CONDUCT OF THE

EXPERIMENT (Should be written on the data sheet for each of the
experiment)
VIII. DISCUSSION OF RESULTS (This should contain the problem/s
encountered in the conduct of the experiment and what remedy/ies have
been applied to solve this/these problem/s)
IX. ANSWERS TO QUESTIONS
X. REFERENCE/S USED

NOTE: It is expected that the preparation of laboratory report

SHOULD ROTATE among the members of the group. An ASTERIX
should mark the name of the student/s who prepared the report.

LABORATORY GRADING SYSTEM

Individual as well as group grades will be generated from the:
Laboratory performance 30%
Individual 10%

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 Group 20%
Products submitted 5%
Examinations 40%
Periodical exams 20%
Quizzes 20%
Reports submitted 10%
Research on a synthesis 10%
Compilation of reports 5%

Total 100%

TABLE OF CONTENTS

TITLE PAGE

FOREWORD

PROPER CONDUCT OF STUDENTS IN THE LABORATORY. . . . . . . . . . . .

........ 3

LABORATORY HAZARDS
OVERVIEW . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5

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WHAT TO DO IN CASE OF AN
EMERGENCY . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10

FORMAT OF LABORATORY
REPORT. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12

LABORATORY GRADING
SYSTEM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13

Experiment 1. EVALUATION AND TESTING OF ORGANIC COMPOUNDS .

...... 15

Experiment 2: SYNTHESIS OF BENZOIC

ACID . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21

Experiment 3: SYNTHESIS OF SULFANILAMIDE . . . . . . . . . . . . . . . . . .

. . . . . . . . 25

Experiment 4: SYNTHESIS OF ASPIRIN

Experiment 5: SYNTHESIS OF METHYL SALICYLATE

Experiment 6: DIAZOTIZATION AND SYNTHESIS OF

AZO DYE, ORANGE II DIHYDRATE.

Experiment 7: SYNTHESIS OF METHYL ORANGE

Experiment 8: SYNTHESIS OF AMYL ACETATE

Experiment 9. SYNTHESIS OF AZELAIC ACID FROM CASTOR OIL

Experiment 10. RESEARCH ON SYNTHESIS OF

ORGANIC COMPOUNDS FROM NATURAL PRODUCTS

Experiment 1. EVALUATION AND TESTING OF ORGANIC

COMPOUNDS

I. Introduction

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 Organic compounds can be simply described as compounds consisting
mainly of the elements, carbon and hydrogen. However, there are some
other elements that can be incorporated into a chemical structure of an
organic compound such as oxygen and nitrogen, thus, producing a vast array
of organic materials that are varying in their physico-chemical properties and
characteristics. These physic-chemical characteristics can be used as a
means to identify them, if not, to characterize them apart from the other
existing chemical compounds.

Classification of organic compounds vary, depending on how they are

chemically structured. Examples of which are organic acids, having the
general formula of RCOOH; alcohols, ROH; esters, RCOOR; and ethers, ROR.
Depending on their chemical classification, the presence of these organic
compounds can be ascertained by evaluating their physical attributes and by
using standard chemical tests to identify their presence.

II. Objectives

The objectives of this experiment are to:

1. familiarize the students with the different types of organic

compounds;
2. differentiate organic compounds in terms of their intrinsic properties;
3. differentiate organic compounds in terms of their behavior towards
ignition;
4. characterize them using chemical tests; and
5. enable students to gain skills in evaluating organic materials.

III. Materials and equipment

Test tubes Test tube rack Test tube holder

Spot plate Evaporating dish Calibrated dropper
Burner Mortar and pestle Litmus papers (blue and red)
pH paper Personal protective equipment (PPE)

Solvents to use: Water

5% Sodium hydroxide
5% Sodium bicarbonate
5% Hydrochloric acid

Chemicals to evaluate: 1. Ethanol

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 2. Phenol
3. Ethyl acetate
4. Acetone
5. Formaldehyde
6. Toluene
7. Diethylether
8. Methanol
9. Salicylic acid
10. Benzoic acid

Equipment: Melting point apparatus

IV. Special instructions

Students are expected to take precautionary measures in handling the
organic compounds to be tested and reagents to be used in this experiment.
Students are NOT ALLOWED TO TASTE sample materials unless
directed.
Students are requested to observe EXTREME CAUTION when handling
chemicals.

V. Procedure
A. Evaluation of the physical state of the sample.
1. Observe the physical state of the sample at room
temperature.
2. Note the color of the sample.
3. Describe the odor of the sample.

B. Solubility property determination.

FOR LIQUID SAMPLES.
1. Calibrate a dropper to 3 ml volume.
2. Take 10 drops of each of the liquid samples to be tested and
place them separately in clean and dry test tubes numbered as the number
of the samples to be tested.
3. Drop by drop, add water by using the dry calibrated dropper.
4. Note any change in color, warming effect, effervescence.
Describe the solubility of the sample in the solvent used, depending on
the number of drops of the solvent used to solubilize the sample being
evaluated.
5. Describe whether the sample was miscible, slightly miscible
and immiscible.

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 6. If insoluble, repeat steps 2-5, this time using 5% NaOH as
solvent.
7. If soluble in 5% NaOH, repeat steps 2-5, this time using 5%
NaHCO3 as solvent.
8. If insoluble in 5% NaOH, repeat steps 2-5, this time using 5%
HCl as solvent.

FOR SOLID SAMPLES.

1. Take 100 milligrams of each of the sample solids for testing
and separately grind them well using a mortar and pestle.
2. Put the ground samples separately in a clean and dry test
tube.
3. Drop by drop, add 5% HCl by using the dry calibrated dropper.
4. Note any change in color, warming effect, effervescence.
Describe the solubility of the sample in the solvent used, depending on
the number of drops of the solvent used to solubilize the sample being
evaluated.
5. Describe whether the sample was very soluble, soluble,
slightly soluble or insoluble.
6. If insoluble, repeat steps 2-5, this time using 5% NaOH as
solvent.
7. If soluble in 5% NaOH, repeat steps 2-5, this time using 5%
NaHCO3 as solvent.
8. If insoluble in 5% NaOH, repeat steps 2-5, this time using 5%
HCl as solvent.

C. Reaction with litmus paper.

If the compound tested is soluble in water, test the nature of the
compound using red and blue litmus papers. Record the reactions observed.

D. Reaction with pH paper.

If the compound tested is soluble in water, test the nature of the
compound using pH paper. Record the reactions observed.

E. Ignition test.
Place 10 drops of the liquid sample in a small evaporating dish
and apply a lighted match to it. Do the same for the samples, except
that they should be introduced to a lighted Bunsen burner.
Observe if the sample tested is flammable or not.

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 Observe the color of the flame and the presence or absence of
soot.
Observe also the burning time.

IDENTITY TESTS
1. Ethanol
a. Mix 5 drops in a small beaker with 1 ml of KMnO4 T.S. Add 5
drops of 2N Sulfuric acid and cover the beaker immediately with filter
paper moistened with a solution recently prepared by dissolving 100
milligrams of Sodium nitroferricyanide and 250 milligrams piperazine in
5 ml water: an intense blue color is produced on filter paper, the
color becoming paler after a few minutes.

b. To 5 ml solution of (I in 10), add 1 ml of 0.1N NaOH then slowly

(over a period of 3 minutes) add 2 ml of 0.1 N Iodine: odor of iodoform
develop, and a yellow precipitate is formed within 30 minutes.

2. Phenol (USP 30)

CAUTION! Avoid contact with the skin, since serious burns
may result.

a. To a solution, add Bromine T.S. drop by drop: a white

precipitate is formed dissolving at first, but becomes
permanent as more of the reagent is added.

b. To 10 ml of the solution (1 in 100), add a drop of Ferric chloride

T.S.: a violet color is produced.

3. Ethyl acetate (NF 25)

It is easily volatilized even at low temperatures and is flammable;
when burned, a yellow flame and an acetous odor is produced.

4. Acetone
Phenylhydrazone test. All aldehydes and ketones readily form
bright-yellow to dark-red 2,4-dinitrophenylhydrazones. Yellow derivatives
are formed from isolated carbonyl groups and orange-red to red derivatives
from aldehydes or ketones conjugated with double bonds or aromatic
rings.
Mix 5 ml of the test sample, acetone, in 0.5 mL of ethanol, and
then add 0.75 mL of 2,4- dinitrophenylhydrazine reagent. Mix

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thoroughly and let sit for a few minutes. A yellow to red precipitate
is a positive test.

Reagent: Dissolve 1.5 g of 2,4-dinitrophenylhydrazine in 7.5 mL of

concentrated
sulfuric acid. Add this solution, with stirring, to a mixture of 10 mL
of water and 35 mL of ethanol.

Sodium nitroprusside Test. In a test tube, place 3 drops of acetone

and 3 ml of water. Then add a drop of 3% NaOH. Add a few drops of
glacial acetic acid and observe carefully the reaction.

5. Formaldehyde
Perform Phenylhydrazone Test using formaldehyde as the test sample.

6. Toluene
Bromine Test
a. One ml of toluene is added to a clean and dry test tube.
b. A few iron filings is added to another test tube, followed by
one ml of toluene to rinse down any iron filings stuck on the test tube
walls.
c. To each test tube was added 3 drops of bromine.
d. The test tubes were placed in a beaker of warm water for 15
minutes. The color of each test tube was observed, and whether or not
HBr was evolved; and the results recorded.

Bayer’s Test (use of aqueous KMnO4)

a. Five ml of KMnO4 solution is placed in one test tube.
b. Five drops of toluene is added to the test tube.
c. The test tube was shaken well for 1-2 minutes and the result
noted.

7. Methanol
CAUTION! Methanol is poisonous.
Use identity tests for ethanol.

8. Isopropyl alcohol

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 9. Benzoic acid (USP 32)
Prepare a saturated solution of benzoic acid in water, and filter
twice. Take 5 ml portion of the filtrate and place it in a dry and clean test
tube. Add Ferric chloride T.S.: A salmon-colored precipitate is
formed.
To a separate 10-mL portion of the filtrate, add 1 mL of 7 N
sulfuric acid and cool the mixture: A white precipitate forms in 10 min;
this precipitate is soluble in ether.

10. Salicylic acid (USP 30)

It meets the requirements for the tests for salicylates:
a. In moderately dilute solutions of salicylates, Ferric chloride T.S.
produces a violet color.
b. The addition of acids to a moderately concentrated solutions
of salicylates produces a white, crystalline precipitate of salicylic
acid that melts between 158° and 161°C.

VI. Data and results

Use Table 1 for preparing your pre-lab report starting with ethanol. Use
additional rows for the rest of the compounds for testing.

Table 1. COMPARATIVE INVESTIGATION OF SOME ORGANIC COMPOUNDS

COMPOUNDS ATTRIBUTES THEORETICAL ACTUAL DATA
DATA
1. Ethanol and so Appearance
on. Color
Odor
Melting point
Boiling point
Solubility in:
Water
5% NaOH

5% NaHCO3

5% HCl

VII. Answer to questions

1. What is the USP definition of solubility?

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2. What is the USP definition of miscibility?
3. Draw a diagram to show the solubility classification of organic compounds.
4. Show the corresponding chemical reactions for the tests for ID performed
on the sample compounds.
IX. References used
http://opencourseware.kfupm.edu.sa/colleges/cs/chem/chem303/files%5C3-
Lecture_Notes_CHEM-303_(Chapter_5).pdf
NF 25/ USP 30
USP 32

Experiment 2. SYNTHESIS OF BENZOIC ACID

I. Introduction
Benzoic acid is a colorless, crystalline solid also known as
benzenecarboxylic acid. It is the simplest aromatic carboxylic acid, with a
carboxyl group (-COOH) bonded directly to the benzene ring. It is found
naturally in the benzoin resin of a number of plants. Benzoin comes from the
bark of a number of balsams of the Styrax genus, most notably Styrax
benzoin and Styrax benzoides. S. benzoin is native to Southeast Asia, and
was traded between Indonesia and China as early as the 8th century c.e.
Benzoin was used for fragrances, spices, medicines, and incense. Today most
benzoin comes from Sumatra (Indonesia) and Laos. Healthy trees do not
produce benzoin, but an incision or wound injuring the cambium results in its
secretion.

CHEMICAL NAME = benzoic acid

CAS NUMBER = 65–85–0
MOLECULAR FORMULA = C7H6O2
MOLAR MASS = 122.1 g/mol
COMPOSITION = C(68.9%) H(4.9%) O(26.2%)
MELTING POINT = 122°C
BOILING POINT = 249°C
DENSITY = 1.3 g/cm3

II. Objectives

This experiment aims to:

Page 22 of 52
 1. synthesize benzoic acid by oxidizing toluene with potassium
permanganate; and
2. characterize the resulting product using identified physico-
chemical procedures

III. Materials and equipment

Erlenmayer flask graduated cylinder beaker

Distilled water 30-40% HCl Toluene
Oxalic acid KMnO4 pH paper
Sandbath plastic basin water pump

IV. Special instructions

Ice-cold water should be running on the condenser of the reflux set-up.

Temperature must be controlled well, since over-heating might cause
EXPLOSION.
Porous boiling chips must be placed on the reaction flask.
Do not leave your set-up unattended.

V. Procedure
A. Synthesis of the compound
A mixture of 5.0 ml toluene, 200 ml water and 8.6 grams of fine,
potassium permanganate powder are placed in a 500 ml round-bottomed
flask equipped with a condenser. The resulting mixture was allowed to
undergo reflux for 4 hours with the use of a sandbath. CAUTION! Boiling
chips made up of small pieces of broken ceramics should be added to the
mixture to even out boiling thus preventing bumping.

Page 23 of 52
 Fig. 1 Reflux set-up. (The heat source is a sand bath, placed on top of a
hotplate.)

Once the reaction is completed, the solution above the formed

manganese dioxide precipitate must be colorless. If not, discoloration of the
solution may be achieved by adding 1 ml alcohol or 0.5g oxalic acid while
heating. This will quickly reduce the remaining potassium permanganate in
the mixture.

The resulting solution is then filtered by suction or through a coffee

filter and the obtained precipitate washed with hot water. The resulting
filtrate is placed in a suitable beaker and the filtrate evaporated to about 50-
100 ml with the use of a water bath and again filtered to remove the newly
formed manganese dioxide. Wash the obtained solid with 5 ml of hot water
and cool the combined filtrate to room temperature. The combined filtrate
was acidified with concentrated HCl until it produces an acidic reaction to pH
paper. The acidity of the reaction mixture will allow the formation of benzoic
acid crystals from solution.

Page 24 of 52
 The mixture is filtered by suction and the crystalline material washed
with cold water. The washed crystals are allowed to dry in-between layers of
filter paper and finally weighed to determine the actual yield of the product.

Fig. 2. The chemical equation involved in the synthesis of benzoic acid

B. Identity tests for benzoic acid

1. Prepare a saturated solution of benzoic acid in water, and filter
twice. Take 5 ml portion of the filtrate and place it in a dry and clean test
tube. Add Ferric chloride T.S.: A salmon-colored precipitate is formed.
2. To a separate 10-mL portion of the filtrate, add 1 mL of 7 N
sulfuric acid and cool the mixture: A white precipitate forms in 10 min; this
precipitate is soluble in ether.

VI. Data and results – Accomplish the Report Sheet.

REPORT SHEET

Experiment No. and Title

Name of the student/s: Date

performed:
Section and Group No.: Date
submitted:

Theoretical Data Actual Data

Page 25 of 52
Physical appearance
Melting point/Boiling
point (as the case
maybe)
Solubility
Yield
Percent yield
Tests for Identity

VI. Qestions to be answered:

1. What is the mechanism of reaction involved in the preparation of

benzoic acid?
2. Give at least five (5) physico-chemical parameters used to ascertain
the identity of benzoic acid. Include chemical reactions (if there is any)
3. Identify the uses of benzoic acid.
4. Provide plant sources where benzoic acid can be found.
5. Give at least five official preparations where benzoic acid is an integral
part of. For what are the named preparations used for. State where these
named preparations are official.

VII. Reference used

Myers, R. L. (2007) The 100 Most Important Chemical Compounds: A
Reference Guide.
Greenwood Press: London.

Experiment. 3 THREE-STEP SYNTHESIS OF SULFANILAMIDE

I. INTRODUCTION

Sulfa drugs were discovered in the early 1900’s and were found
to be active as antibacterial agents. Sulfanilamide inhibits the

Page 26 of 52
 formation of folic acid in bacteria, thus inhibiting its growth and
development due to non-formation of THFA (tetrahydrofolic acid), an
essential compound in the development of the bacteria.

Sulfanilamide is the common name for p-

aminobenzenesulfonamide. The term sulfonamide indicates that it is
an amide of sulfanilic acid. Sulfanilic acid (precursor of sulfonamide)
can be prepared from aniline and sulfuric acid, and the name
implies an aniline unit to which has been attached a sulfonic acid
group. The names of the intermediates involved in the synthesis
starting with the chlorosulfonation of acetanilide are all based from
the parent compound.

Sulfanilamide is a white, crystalline compound, having a

molecular formula of C6H8N2SO2 used in the treatment of bacterial
infections. The term sulfanilamide is also used to describe a family
of molecules containing this functional group as exemplified by
furosemide, a diuretic; sulfadiazine and sulfamethoxazole, for the
treatment of urinary, respiratory and GIT infections.

Sulfanilamide is easily synthesized from aniline in four steps:

II. OBJECTIVE : To synthesize sulfanilamide from aniline.

III. INSTRUCTIONS:
A. Read on the physical properties (physical appearance and
melting point) of the intermediates and the product.
B. Look for chemical tests that would identify the intermediates and
the product.
C. Read on techniques for heating, vacuum filtration, decolorization,
crystallization and recrystallization, purification and melting point
determination.

Page 27 of 52
 D. Compute for the amounts (in grams or milliliters) of other
reactants and reagents based on the amount of starting material
that is assigned.
E. At the end of the laboratory period, pass a datasheet complete
with the computations of amounts of reactants, validated
amounts, and actual results of physical and chemical tests.

IV. REAGENTS : Aniline Conc. HCl

Sodium acetate Chlorosulfonic acid

Conc. HNO3 Sodium bicarbonate

Distilled water Acetic anhydride

Conc. Ammonia Ice water

V. REAGENT INFORMATION AND PRECAUTIONS:

COMPOUND CHARACTERISTICS
Conc. HCl Irritant, corrosive
Conc. Ammonia Hazardous gas, caustic and corrosive
Aniline Irritant to the eyes and skin, harmful if
ingested and inhaled and a possible
carcinogen
Sodium acetate Irritant to the eyes and skin
Chlorosulfonic acid Corrosive and reacts violently with water
Sodium bicarbonate Irritant especially to respiratory system

VI. PROCEDURE:
A. Acetanilide
1. Dissolve 1 gram of aniline in 30 ml of distilled water and I ml
of conc. HCl in a 125 ml Erlenmayer flask. If the solution is
colored, vacuum filter through decolorizing charcoal.
2. Measure out 1.2 ml of acetic anhydride and prepare a solution
of 1 gram of sodium acetate in 6 ml of distilled water.
3. Add the acetic anhydride to the solution of aniline with
stirring, and at once add the sodium acetate solution.
4. Stir the mixture, cool it in ice, and collect and weigh the
product acetanilide.
NOTE: The material needs to be completely dry before the
next step.
Characterize the acetanilide.

Page 28 of 52
B. p-Acetamidobenzenesulfonyl chloride
1. Place 0.5 gram dry acetanilide in a dry 25 ml Erlenmayer
flask.
2. Add 1.25 ml of chlorosulfonic acid(Org13) (WARNING!
Corrosive and reacts violently with water) a few drops at
atime using a Pasteur pipette (CAUTION! No metal needles!).
3. After about 10 minutes, the reaction should subside and
almost all of the acetanilide should have dissolved.
4. Heat the mixture in a hot water bath for about 10 minutes to
complete the reaction.
5. Pipet the mixture slowly with stirring into 7 ml of ice water in
another 25 ml Erlenmayer flask (WARNING! USE EXTREME
CAUTION WHEN DOING THIS!)
6. Rinse the reaction flask with cold water and stir the product
until an even suspension of white solid is obtained.
7. Vacuum filter the p-acetamidobenzenesulfonyl chloride and
wash it with water.

C. p-Acetamidobenzenesulfonamide
1. Transfer the solid (you can use the solid even if it is wet) to
the rinsed 25 ml Erlenmayer and add 1.5 ml of concentrated
ammonia and 1.5 ml of distilled water.
2. Heat the mixture to just below the boiling point on a hot plate
with occasional swirling for 5 minutes.
3. Cool the mixture in an ice bath and collect the p-
acetamidobenzenesulfonamide by suction filtration and allow
it to drain thoroughly.

D. Sulfanilamide
1. Transfer the moist solid to a 25 ml Erlenmayer.
2. Add 0.5 ml of conc. HCl and 1 ml of water.
3. Boil the mixture gently until the solid dissolves and then
continue heating at the boiling point for about 10 minutes
longer (do not evaporate to dryness).
4. Cool the solution to room temperature. No solid should
deposit. If the solid material is seen, continue heating
for a little while longer.
5. To the cool solution, add a saturated aqueous solution of 0.5
grams of sodium bicarbonate until the solution is neutral to
pH paper.
6. Cool the mixture in ice and vacuum filter the sulfanilamide.
Characterize the product.

Page 29 of 52
 7. Calculate the percentage yield of your product.

REPORT SHEET

Experiment No. and Title

Name of the student/s: Date

performed:
Section and Group No.: Date
submitted:

Starting material: ____________ g

THEORETICAL DATA ACTUAL DATA

Physical appearance

Melting point

Solubility

Yield

Percent yield Assuming 100% yield

Test for sulfanilamide

VII. Computations:
1. Determine the theoretical yield of the intermediate and the
product in your preparation.
2. Calculate the percentage yield of your product using the formula:

Percentage yield = Actual yield x 100

Theoretical yield

VIII. Questions to be answered:

Page 30 of 52
 1. What is the mechanism of reaction involved in the preparation of
sulfanilamide?
2. How are antibiotics usually prepared/obtained from natural
sources?
3. Provide chemical derivatives of sulfanilamide and for what are
these derivatives used.

Experiment 4: SYNTHESIS OF ASPIRIN

I. INTRODUCTION

Historically, the salicylates were among the first drugs to achieve

recognition as analgesics. In the 17 th century, the extract of willow,
Salix spirea, was found to produce fever-reducing properties and
was used by Jesuit missionaries (Bailey, Jr. and Bailey 1998). In
1827, Leroux isolated salicin (I), the active principle in the willow
bark and Piria, in 1838, prepared salicylic acid (II) from salicin. By
1860, Kolbe and Lautermann prepared it synthetically from phenol
(Willette 1991). Although salicylic acid is an effective antipyretic, it
causes severe stomach irritation in some people and for this reason
the research for a pain reliever continued in the late 1800s (Bailey,
Jr. and Bailey 1998). Aspirin, chemically known as acetylsalicylic
acid, the salicylate ester of acetic acid, was first prepared in 1853
by Gerhardt, but remained obscure until Felix Hoffmann, who
worked for the Bayer Company, discovered its pharmacologic
activities in 1899 when he used the acetyl derivative on his father
suffering from arthritis. The name aspirin comes from spirin, an old
name of salicylic acid or spiric acid, derived from its natural source
of spirea plants (Willette 1991). Unfortunately, even aspirin causes
stomach distress in some individuals and minor, usually clinically
unimportant, gastric or intestinal bleeding (Bailey, Jr. and Bailey
1998). Nevertheless, aspirin remains, until now, the most popular
and the most versatile drug ever to have been synthesized.

Salicylates, in general, exert their antipyretic action in febrile

patients by increasing heat elimination of the body through
mobilization of water and consequent dilution of the blood. This
brings about perspiration, causing cutaneous dilatation. This does
not occur with normal temperatures. The antipyretic and the
analgesic actions are believed to occur in the hypothalamic area of
the brain. Thye antitheumatic and antithrombotic actions of aspirin
is said to arise from its selective action on the synthesis of the

Page 31 of 52
 prostaglandin-related thromboxane and prostacyclin, which are
counterbalancing factors involved in platelet aggregation and are
released when tissue is injured (Willette 1991).

Salicylcic acid is a bifunctional compound, having both a phenolic

hydroxyl group and a carboxyl group at ortho positions of each
other. Hence, it can undergo two different types of esterification
reactions, acting either as the alcohol or the acid partner in the
reaction. In the presence of a carboxylic acid or acid derivative, e.g.
acetic anhydride, salicylic acid acts as the alcohol forming, for
example, acetylsalicylic acid; whereas in the presence of alcohols,
e.g. methanol, it forms esters such as methyl salicylate through its
carboxylic group. Because salicylic acid has both the phenolic
hydroxyl group and the carboxy of the product aspirinl group,
polymers often complicate the esterification reaction. Fortunately,
acetylsalicylic acid will react with sodium bicarbonate. This
difference in the behavior will be utilized for the purification of the
product aspirin (Pavia et al. 1976).

The most likely impurity in the final product is salicylic acid itself,
which can arise from incomplete acetylation or from hydrolysis of
the product during the isolation steps. This material is removed
during the various stages of the purification and in the final
crystallization of the product. Salicylic acid, like most phenols, forms
a highly colored complex with ferric chloride. Aspirin, which has this
group acetylated, will not give the color reaction. Thus, the
presence of this impurity in the final product is easily detected
(Pavia et al. 1976).

A simple method of the identification of aspirin is based on the

principle that aspirin forms a beautifully colored blue complex with
cupric acetate (Wilcox, Jr. and Wilcox 1995).

II. OBJECTIVES:
A. To demonstrate the process of esterification reaction;
B. To understand electronic and steric effects in esterificaton
reactions;
C. To utilize the differing rates of reactions and differences in
solubilities to optimize the yield of the desired product; and
D. To utilize chemical tests in determining the presence of functional
groups.

Page 32 of 52
III. INSTRUCTIONS:

A. Read on the physical properties (physical appearance and

melting point).
B. Look for the theoretical results of each intermediate/product in
the chemical tests given and read on the principle involved in
each test.
C. Read on techniques for heating, vacuum filtration, decolorization,
crystallization and recrystallization, purification and melting point
determination.
D. Compute for the amounts (in grams or milliliters) of other
reactants and reagents based on the amount of starting material
that is assigned.
E. Prepare a schematic diagram for the procedure.
F. Draw the set-up necessary in the procedure. Label completely.
G. At the end of the laboratory period, pass a datasheet complete
with the computations of amounts of reactants, validated
amounts, and actual results of physical and chemical tests.

IV. REAGENTS: Salicylic acid Benzene

Acetic anhydride Petroleum ether (optional)
Conc. Sulfuric acid 1% Ferric chloride
Saturated NaHCO3 Cupric acetate
Conc. HCl 95% Ethanol
V. PROCEDURE:

1. Weigh out 3.0 grams of salicylic acid and place it in a dried,

250 ml Erlenmayer flask.
2. Measure 6.0 ml of acetic anhydride and add this to your flask.
Be sure tho do this in the hood and wear your goggles.
Don’t let the acetic anhydride be in contact with your
skin and don’t let the vapors be in contact with your
eyes.
3. Carefully add 5 to 10 drops of 85% phosphoric acid, a
catalyst, to the flask and swirl to mix everything thoroughly.
4. Still in the hood, heat the mixture for about 10 minutes in a
beaker of warm water (70-80 degrees)
5. After heating, cautiously add 20 drops of distilled water.
6. Remove the reaction set-up from the hood and then add 20 ml
distilled water and cool in an ice bath. If crystals do not
appear, scratch the walls of the flask with a stirring rod to
induce crystallization. As soon the crystalline nuclei has

Page 33 of 52
 appeared, stop the process and let he crystalline material
form undisturbed.
7. Once crystallization is complete, filter the solid aspirin through
a pre-weighed filter paper using a Buchner funnel. Wash the
crystals with 2-3 ml of chilled water. The filtrate is mostly
water and can be washed down the sink. Allow filtration to go
through for about 15 minutes. Test for the presence of
unreacted salicylic acid using the ferric chloride test below
(Pavia et al. 1976). If the sample tests positive (violet colored
solution), the crude aspirin is subjected to the recrystallization
procedure as follows:

Dissolve a small sample of the final product in a minimum

amount of hot benzene, while gently and continuously heating
the mixture on the steam bath. Caution: Benzene is a
potential carcinogen. Handle it with great care. If any
solid a remains, filter the solution by gravity through a fluted
filter placed in a short-stemmed funnel w2hich has previously
been pre-heated by pouring hot benzene through it. Let the
filtered solution stand. On cooling to room temperature, the
aspirin should crystallize. If it does not, add a little petroleum
ether and cool the solution slightly in ice water, while
scratching the inside of the glass with a glass rod. Collect the
product by vacuum filtration. Repeat the ferric chloride test.

8. Place the filter paper with the product in a watch glass and
place it inside an oven at 100 degrees Centigrade for about
30 minutes until it is dry.
9. Put the dry aspirin and the filter paper into an appropriate,
pre-weighed container and weigh again.
10. Calculate the weight of your final product. Determine the
percentage yield of your product.

CHEMICAL TESTS

Ferric chloride test. To a pinch of the crude product,

add 1% ferric chloride solution and shake vigorously until the
product is dissolved. Observe any changes in the color of the
solution (Pavia et al. 1976)

Hydroxamic acid test. Dissolve a small amount of

the ester in a mixture of 1 ml of 0.5 N Hydroxylamine . (dissolved

Page 34 of 52
 in 95% ethanol) and 0.2 ml of 6N Sodium hydroxide. Heat the
mixture to boiling for a few minutes. Cool the solution and then
add 2 ml of 1N Hydrochloric acid. If the solution becomes cloudy,
add 2 ml of 95% ethanol to clarify it. Add a drop of 5% ferric
chloride solution and note whether a color is produced. If the
color fades, continue to add ferric chloride until it persists. A
positive test should give a deep burgundy or magenta color
(Pavia et al. 1976).

Test for aspirin. In a 50 ml beaker, prepare a solution of

0.05 g of cupric acetate monohydrate in 10 ml of water that has
been heated to about 55 degrees. In a 20 ml test tube dissolve
0.07 grams of acetylsalicylic acid in 1.25 ml of 95% ethanol and
add the solution to the warm solution of cupric acetate. Stir the
mixture well and set the test tube aside to cool slowly. Observe
the formation of dark blue precipitate of cupric acetylsalicylate or
cupric aspirinate (Wilcox, Jr. and Wilcox 1995).

VI. DATA AND RESULT:

REPORT SHEET

Experiment No. and Title

Name of the student/s: Date

performed:
Section and Group No.: Date
submitted:

Starting material: ____________ g

THEORETICAL DATA ACTUAL DATA

Physical appearance

Melting point

Page 35 of 52
Solubility

Yield

Percent yield Assume 100% yield

Test/s for aspirin

VII. QUESTIONS TO ANSWER:

1. What is the purpose of the concentrated acid in the acetylation
reaction?
2. Which would you expect to be a stronger acid: benzoic acid,
salicylic acid, or aspirin? Explain.
3. What is the purpose of concentrated acid in the purification of
aspirin?
4. How do the techniques of scratching and seeding induce
crystallization?
5. How is the ferric chloride test used to detect the presence of
salicylic acid? What general type of compound gives a positive
result for this test?
6. Can you use water as solvent for recrystallization? Explain.

VIII. REFERENCES:

Bailey, P.S., Jr. and Bailey, C.A., 1998. Organic Chemistry: A Brief
Survey of Concepts and Application. 5th Edition. Singapore: Prentice-
Hall Internaional, 393-394.

Pavia, D.L., Lampman, G.M., and Kriz, G.S., Jr., 1976. Introduction to
Organic Laboratory Techniques: A Contemporary Approach>
Philadelphia: W.B. Saunders, 25-30, 423.

Page 36 of 52
 Wilcox, C.F., Jr. and Wilcox, M.F., 1995. Experimental Organic
Chemistry: A Small Scale Approach. 2nd Edition. New Jersey:
Prentice-Hall, 486-487.

Willette, R.E., 1991. Analgesic Agents. In: J.N. Delgado and W.A.
Remers, ed. Wilson and Gisvold,s Textbook of Organic Medicinal and
Pharmaceutical Chemistry. 9th Edition. Philadelphia: J.B. Lippincott,
657.

Experiment 5: SYNTHESIS OF METHYL SALICYLATE

I. INTRODUCTION

Methyl salicylate, also known as salicylic acid methyl ester, oil of

wintergreen, betula oil, or methyl-2-hydroxybenzoate, is a natural
product which can be obtained from several plant species. Some of
the plants producing it are called wintergreens, hence its common
name. Plants containing methyl salicylate produce this organic ester
most likely as an anti-herbivore defense. If the plant is infested with
herbivorous insects, the release of methyl salicylate may function as
an attractant of beneficial insects that could act as natural

Page 37 of 52
 predators for the herbivorous pests. Numerous plants produce the
compound in very small amounts.

It is used as a rubefacient in deep heating liniments, and in small

amounts as a flavoring agent at no more than 0.04%. It is also used
to provide fragrance to various products and as an odor-masking
agent for some organophosphate pesticides. If applied in high
quantities, it can cause stomach and kidney problems.

It is one of the compounds that is attractive to males of various

species of orchid bees, who apparently gather the chemical to
synthesize pheromones; it is commonly used as a bait to attract and
collect these bees for study.

Methyl salicylate also has the ability to clear plant or animal

tissue samples of color, and as such is useful for microscopy and
immunochemistry when excess pigments obscure structures or
block light in the tissue being examined. This clearing generally only
takes a few minutes, but the tissue must first be dehydrated in
alcohol.

In pure form, methyl salicylate is toxic, especially when taken

internally. The lowest published lethal dose is 101 mg/kg body
weight in adult humans. It has proved fatal to small children in
doses as small as 4 ml.

Methyl salicylate can be produced by esterifying salicylic acid

with methanol.

II. OBJECTIVES:
1. To synthesize methyl salicylate by the process of esterification.
2. To know the requisites for the esterification reaction to occur.

III. REAGENTS: Salicylic acid

Methanol
Conc. Sulfuric acid
IV. REAGENT INFORMATION:

COMPOUND CHARACTERIISTIC
Salicylic acid Irritant to the eyes and skin
Methanol Toxic, CNS depressant
Sulfuric acid Highly corrosive chemical. May
cause burns.

Page 38 of 52
 V. PROCEDURE:
Place 1.0 gram of salicylic acid and 6.0 ml of methyl alcohol in a
25 ml Erlenmmayer flask. Add 4.0 drops of concentrated sulfuric
acid (REMINDER! Note the odor of the resulting solution) and then
place the test tube in a water bath previously heated to 7o degrees
and heat the solution for 15 minutes.

NOTE: The boiling point of methanol is 64.6 degrees, care should

be taken to avoiod overheating and minimize bumping.

Allow the resulting product to cool to room temperature and note

the odor before placing it in a suitable container and properly label
the product.

VI. DATA AND RESULT

REPORT SHEET

Experiment No. and Title

Name of the student/s: Date

performed:

Page 39 of 52
Section and Group No.: Date
submitted:

Starting material: ____________ g

THEORETICAL DATA ACTUAL DATA

Physical appearance

Melting point

Solubility

Yield

Percent yield Assume 100% yield

Test/s for aspirin

VII. QUESTIONS TO ANSWER:

1. Give the general reaction for the synthesis of methyl salicylate.
2. Propose a reaction mechanism for this synthesis.
3. Provide the physico-chemical characteristics of the product.
4. How can the product be tested by chemical means? Include
pertinent chemical reactions.
5. What physico-chemical methods can be resorted to, to identify
methyl salicylate? Provide the characteristic IR and NMR spectra
for this compound.

Experiment 6. DIAZOTIZATION AND SYNTHESIS OF AZO DYE, ORANGE

II DIHYDRATE Experiment 7. SYNTHESIS OF METHYL ORANGE

Page 40 of 52
I. INTRODUCTION

Azo dyes, the most commonly employed of all kinds of dye can
be prepared by azo coupling. This is due to the fact that aryl
diazonium ions (ArN N+) are weak electrophiles and give strongly
colored compounds (ArN=Nar’) by electrophilic substitution, but
only on aromatic rings which are strongly activated by hydroxyl or
amino groups. Most dyeing operations utilize hot aqueous solutions
of the dyes, many of which bear sulfonic acid groups to improve
their water solubility. Two dyes will be prepared in this experiment.
Sulfanilic acid will be diazotized by reaction with nitrous acid, and
the resulting diazonium-sulfonate inner salt will then be coupled
with 2-naphthol and N,N-dimethylaniline to give the dyes commonly
known as Orange II and Methyl orange, respectively.

II. REAGENTS:

For diazotization and synthesis of Orange II Dihydrate:

Sulfanilic acid monohydrate 2.5% aq. sodium carbonate

Sodium nitrite Ice
Concentrated HCl 2-Naphthol
10% aq. NaOH NaCl
Distilled water Ethanol

For the synthesis of Methyl Orange

N,N-dimethylaniline Glacial acetic acid

10% aq. NaOH Saturated aq. NaCl

III. REAGENT INFORMATION:

COMPOUND CHARACTERIISTIC
Sodium nitrite Decomposed even by weak acids
with the evolution of brown
fumes
Conc. HCl Irritant; corrosive
N,N-dimethylaniline Poisonous!
Glacial acetic acid Eye irritant

IV. PROCEDURES:

Procedure for diazotization of sulfanilic acid

Page 41 of 52
 Place 2.4 grams of sulfanilic monohydrate in a 125 ml
Erlenmayer flask, and dissolve it in 25 ml of 25% aqueous sodium
carbonate by boiling. Cool the solution in the flask with running
water before adding 0.95 gram of sodium nirite with stirring to
dissolve the solid. Pour the solution into a 50 ml Erlenmayer flask
containing 12.5 grams of ice and 2.5 ml of concentrated
hydrochloric acid. Within a few minutes the white diazonium
sulfonate inner salt should form a suspension. Transfer half of it to a
100 ml beaker, and keep both containers cold until each is used in
later steps.

Procedure for the preparation of Orange II dehydrate

Dissolve 0.9 gram of 2-naphthol in 5 ml of 10% aqueous sodium

hydroxide in a 100 ml beaker. (It may be necessary to warm the
mixture to dissolve the naphthol completely.) Chill the solution to
bellow room temperature before adding the contents of the
Erlenmayer flask containing the diazonium-sulfonate inner salt.
Rinse the flask with a small amount of water, and add this to the
beaker containing the 2-naphthol. The dye, as a sodium salt, forms
rapidly. Stir the resulting paste thoroughly to ensure good mixing
and, after 10 minutes, heat the mixture to dissolve all solids.
Dissolve 2.5 grams of sodium chloride in this solution, heating if
necessary. Then let the solution cool to room temperature
undisturbed. Chill the solution in an ice bath for a few minutes with
gentle stirring. Collect the product on a Buchner funnel. Since the
filtration is slow, pour small portions into the funnel successively;
then rinse the beaker with saturated aqueous sodium chloride.

Transfer the filter cake to a beaker, washing the filter paper and
funnel with a small amount of water (about 12 ml). Then, dissolve
the solid by heating the water at the boiling point, and filter through
a Buchner funnel that has been pre-heated on a steam bath. Pour
the filtrate into a 100 ml Erlenmayer flask, rinsing the filter flask
with 1-2 ml of water. The total volume should not exceed 15 ml.
Cool to 80 degrees before adding 25-30 ml of ethanol; then, set it
aside to crystallize. Before collecting the product, chill the mixture
thoroughly, and wash the crystals on the filter with a small amount
of ethanol. Weigh the product and calculate the yield. (Orange II

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 crystallizes as the dehydrate for which you must allow in the
calculation of the yield.)

Synthesis of Methyl Orange

Thoroughly mix 0.8 ml of N,,N-dimethylaniline and 0.65 ml of

glacial acetic acid in a small test tube. Dd this solution of
dimethylanilinium acetate to the suspension of the
diazoniumsulfonate inner salt in the 100 ml beaker, rinsing the test
tube with a small amount of water. Stir the mixture thoroughly, and
watch the red acid form of the dye separate, such that a stiff paste
results, in 5-10 minutes. Add 9 ml of 10% aqueous sodium
hydroxide to produce the orange sodium salt. Stir well, and heat the
mixture to boiling, at which point most should be dissolved. Place
the beaker in an ice bath and allow it to cool undisturbed.

Collect the product by filtration through a Buchner funnel using

saturated aqueous sodium chloride to rinse the beaker and to wash
the dark mother liquor from the filter cake.

Methyl orange is sometimes used as an acid-base indicator. To

observe the color changes with pH, dissolve a few crystals in a small
amount of water in a test tube, and add a few drops of dilute
aqueous hydrochloric acid and then few drops of dilute aqueous
sodium hydroxide to show that the color changes are reversible.

Determine the percentage yield for both azo dye products.

V. DATA AND RESULT

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 REPORT SHEET

Experiment No. and Title

Name of the student/s: Date

performed:
Section and Group No.: Date
submitted:

Starting material: ____________ g

THEORETICAL DATA ACTUAL DATA

Physical appearance

Melting point

Solubility

Yield

Percent yield Assume 100% yield

Test/s for aspirin

VI. QUESTIONS TO BE ANSWERED:

1. Draw the structure of methyl orange at acidic and basic pH, and
underneath each structure, indicate the color observed for each
compound.
2. What are the methods of fabric dying?
3. What are mordant dyes? Explain how mordant dyes adhere to
fabrics.

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Experiment 8: SYNTHESIS OF AMYL ACETATE

I. INTRODUCTION
Banana oil is the common name of the chemical compound
properly known as amyl acetate also known as isopentyl acetate. It
is a colorless liquid ester derived from amyl alcohol. It resembles
the smell of bananas but is not naturally found in banana fruit.

II. REAGENTS: Isopentyl alcohol Glacial acetic acid

Conc. Sulfuric acid Distilled water
5% Sodium bicarbonateSaturated aq. NaCl solution
Anhydrous magnesium sulfate

III. PROCEDURE:
1. Place 15 ml of isopentyl alcohol in a 100 ml round-bottom flask
and add 20 ml of glacial acetic acid.
2. Swirl the flask and carefully add 4.0 ml of concentrated sulfuric
acid (Caution! Highly corrosive).
3. Attach a reflux condenser and, using a heating mantle, reflux the
mixture for 1 hour (do not forget to add boiling chips). Cool to
room temperature.
4. Place the reaction mixture in a separatory funnel and add 55 ml
of cold water (Remember to rinse the reaction flask with 10 ml of
cold water and add it to the separatory funnel). Separate the
lower aqueous layer.
5. Extract the organic layer (upper layer) with 25 ml of 5% sodium
bicarbonate solution twice (test to be certain that the aqueous
layer is basic to litmus otherwise wash again). CAUTION!
Formation of carbon dioxide which will exert pressure inside the
separatory funnel.
6. Extract the organic layer with 25 ml of water. Finally, add 5.0 ml
of saturated aqueous NaCl to aid in layer separation (it removes
traces of water from the organic layer). Do not shake this
solution but simply swirl. Draw off the lower aqueous layer. Pour

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 the top organic layer into an Erlenmayer flask and dry with 2
grams of anhydrous magnesium sulfate.
7. Decant the ester into the distilling flask (make sure that the
drying agent is excluded and all glassware completely dry). Set
up the distillation apparatus. Be certain that the adapter is open
to the atmosphere and that your thermometer is placed correctly
in the distilling head(do not forget the boiling chips). Collect all
distilled material but collect the fraction between 134 and 143
degrees in a separate tared flask. Keep the receiver flask cold to
reduce the vapor escaping into the lab environment. Never distill
to dryness. Record the barometric pressure in the laboratory.
8. Weigh the product and calculate the percentage yield.

IV. DATA AND RESULT

REPORT SHEET

Experiment No. and Title

Name of the student/s: Date

performed:
Section and Group No.: Date
submitted:

Starting material: ____________ g

THEORETICAL DATA ACTUAL DATA

Physical appearance

Melting point

Solubility

Yield

Percent yield Assume 100% yield

Page 46 of 52
Test/s for aspirin

V. QUESTIONS TO BE ANSWERED:
1. Provide the chemical reaction involved in the synthesis of amyl
acetate.
2. Provide a possible reaction mechanism for the above synthesis to
occur.
3. Give reasons for the following steps:
a. swirling of the flask containing the reactants prior to addition
of sulfuric acid.
b. washing with cold water
c. extraction of the organic layer with 5% sodium bicarbonate
solution twice
d. addition of 5 ml of saturated aqueous NaCl
e. keeping the receiver flask cold
4. What are the uses of banana oil?
5. Is banana plant a tree? Justify your answer.

VI. REFERENCES:
http://bllogcritics.org/scitech/article/what-is-banana-oil/
http://www.umsl.edu/orglab/experiments/Bananaoil.html
http://www.hort.purdue.edu/newcrop/morton/banana.html

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Experiment 9: PREPRATION OF AZELAIC ACID FROM CASTOR OIL

I. INTRODUCTION

Castor oil is a vegetable oil obtained from the castor bean. It is a

colorless to very pale, yellow liquid with mild or no odor or taste.
Castor oil is composed of a mixture of triglycerides, about 75% of
which is tricinolein. The remainder consist of diricinoleoglycerides
with the third acyl group, representing either oleic, linoleic,
dihydroxystearic, or saturated (stearic or palmitic) acid.

Azelaic acid is a saturated dicarboxylic acid found naturally in

wheat, rye and barley. It is a natural substance that is produced by
Melassezia furfur, a yeast that lives on normal skin. It is industrially
produced by the ozonolysis of oleic acid. It is effective against a
number of skin conditions, such as mild to moderate acne, when
applied topically in a cream population of 20%. Azelaic acid may be
useful as a hair growth stimulant, although at present there is no
clinical study to confirm the efficacy. Its antibacterial property
reduces the growth of bacteria in the follicle (Propionibacterium
acnes and Staphylococcus epidermidis). It has a keratolytic and
comedolytic property which normalizes the disordered growth of the
skin cells, lining the follicle. Azelaic acid does not result in bacterial
resistance to antibiotics, reduction in sebum production,
photosensitivity (easy sunburn), staining of skin or clothing, or
bleaching of normal skin or clothing.

II. OBJECTIVE OF THE EXPERIMENT:

To prepare a useful chemical from a natural source.

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III. PLANT SOURCE AND REAGENTS:
Castor oil (unflavored) KOH pellets
95% ethanol Potassium permanganate
Conc. Sulfuric acid

IV. REAGENT INFORMATION:

COMPOUND CHARACTERIISTIC
95% ethanol Toxic, CNS depressant
Conc. Sulfuric acid Highly corrosive chemical. May
cause burns.
KOH pellets Caustic irritant

V. PROCEDURE:

Fifteen grams castor oil is added to a solution of 15 grams

potassium hydroxide in 45 ml 95% ethanol. The mixture is placed in
a 500 ml Erlenmayer flask equipped with a reflux condenser and is
boiled for 30 minutes. Timing starts at the appearance of the first
drop of condensate. The solution is then poured into 100 ml water
and acidified by the addition of 30 ml of 40% sulfuric acid in water.
The procedure is repeated until all the ricinoleic acid have been
liberated. The acid that separates is washed until the aqueous layer
becomes clear. The yield of crude ricinoleic acid thus obtained
approximately 16 grams.

Twelve grams of ricinoleic acid is dissolved in 80 ml water

containing 4 grams potassium hydroxide. In a one liter round-
bottomed flask equipped with a powerful mechanical stirrer are
placed 34 grams of potassium permanganate and 400 ml water at
35 degrees. The mixture is stirred to facilitate solution of the
permanganate, and, if necessary, heat is applied to maintain the
temperature at 35 degrees. When the permanganate has
completely dissolved, the alkaline solution of ricinoleic acid is added
in single portion with vigorous stirring. The temperature rises to
about 75 degrees. Stirring is continued for half an hour, or until a
test portion added to water shows no permanganate color. To the
mixture is now added a solution of 20 g concentrated sulfuric acid in
63 ml water. The acid must be added slowly and carefully to prevent
too rapid evolution of carbon dioxide with consequent foaming. The

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 mixture is heated on a steam bath for 10 minutes to coagulate the
manganese dioxide, which is filtered while still very hot. After
filtration, the manganese dioxide is placed in a 400 ml beaker and
boiled with 50 ml water in order to dissolve any azelaic acid that
may adhere to it. This mixture is filtered while hot, and the filtrate is
added to the main portion. The combined filtrates are evaporated to
a volume of about 200 ml and this solution is cooled in ice. The
crystals that separate are filtered with suction, washed once with
cold water, and dried. The yield is 3-4 grams material of melting
point 95-106 degrees. The crude substance is dissolved in 60 ml
boiling water, filtered with suction, and allowed to cool. The crystals
are filtered, washed with water, and dried, the yield, 2-3 grams;
melting point, 104-106 degrees.

VI. DATA AND RESULT

REPORT SHEET

Experiment No. and Title

Name of the student/s: Date

performed:
Section and Group No.: Date
submitted:

Starting material: ____________ g

THEORETICAL DATA ACTUAL DATA

Physical appearance

Melting point

Solubility

Page 50 of 52
Yield

Percent yield Assume 100% yield

Test/s for aspirin

VII. QUESTIONS TO ANSWER

1. Provide the chemical reactions involved in the preparation of
azelaic acid from castor oil.
2. Propose the mechanism of reaction involved based on the
provided chemical reaction.
3. Characterize castor oil. Provide the botanical origin of the plant
source.
4. Provide the names of 5 plant acids found in castor oil and give
their chemical structures, as well as their IUPAC names.

IX. REFERENCES:

Bailey, P.S., Jr. and Bailey, C.A., 1998. Organic Chemistry: A Brief
Survey of Concepts and Application. 5th Edition. Singapore: Prentice-
Hall Internaional, 393-394.

Pavia, D.L., Lampman, G.M., and Kriz, G.S., Jr., 1976. Introduction to
Organic Laboratory Techniques: A Contemporary Approach>
Philadelphia: W.B. Saunders, 25-30, 423.

Wilcox, C.F., Jr. and Wilcox, M.F., 1995. Experimental Organic

Chemistry: A Small Scale Approach. 2nd Edition. New Jersey:
Prentice-Hall, 486-487.

Willette, R.E., 1991. Analgesic Agents. In: J.N. Delgado and W.A.
Remers, ed. Wilson and Gisvold,s Textbook of Organic Medicinal and
Pharmaceutical Chemistry. 9th Edition. Philadelphia: J.B. Lippincott,
657.

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