Organic Laboratory Techniques and Manuals

For pharmacist students

Debrecen
2009

I. SAFETY GUIDELINES
The introduction of organic laboratory techniques has significantly
lessened many of the dangers found in organic laboratories; however, you
still must take many precautions. Your laboratory instructor will advise you
of the specific rules for your laboratory. The following list of safety
guidelines should be observed in all organic laboratories.
1. Eye Safety
Always Wear Approved Safety Glasses or Goggles. This sort of eye
protection must be worn whenever you are in the laboratory. Even if you are
not actually carrying out an experiment, a person near you might have an
accident that could endanger your eyes, so eye protection is essential. We
recommend that contact lenses not be worn in the laboratory, even when
safety goggles are worn over them. Contact lenses can trap chemicals against
the eye and make it difficult to wash the eye quickly. Soft contact lenses
present an additional hazard because they can absorb harmful chemical
vapors. In case any chemical enters your eyes, wash immediately with water.
2. Fires
Use Care with Open Flames in the Laboratory. Because an organic
chemistry laboratory course deals with flammable organic solvents, the
danger of fire is frequently present. Because of this danger, DO NOT
SMOKE IN THE LABORATORY. Further-more, exercise supreme
caution when you light matches or use any open flame. Always check to see
whether your neighbors on either side, across the bench, and behind you are
using flammable solvents. If so, either delay your use of a flame or move to a
safe location, such as a fume hood, to use your open flame. Many flammable
organic substances are the source of dense vapors that can travel for some
distance down a bench. These vapors present a fire danger, and you should be
careful, since the source of those vapors may be far away from you. Do not
use the bench sinks to dispose of flammable solvents. If your bench has a
trough running along it, pour only water (No flammable solvents!) into it.
The troughs and sinks are designed to carry water from the condenser hoses
and aspiratorsnot flammable materials.
Learn the Location of Fire Extinguishers, Fire Showers, and Fire
Blankets. For your own protection in case of a fire, you should learn
immediately where the nearest fire extinguisher, fire shower, and fire blanket
are. You should learn how these safety devices are operated, particularly the
fire extinguisher. Your instructor can demonstrate how to operate it.
If you have a fire, the best advice is to get away from it and let the
2

instructor or laboratory assistant take care of it. DON'T PANIC! Time spent
in thought before action is never wasted. If it is a small fire in a container, it
usually can be extinguished quickly by placing a wire gauze screen with a
ceramic fiber center or, possibly, a watch glass, over the mouth of the
container. It is good practice to have a wire screen or watch glass handy
whenever you are using a flame. If this method does not take care of the fire
and if help from an experienced person is not readily available, then
extinguish the fire yourself with a fire extinguisher.
Should your clothing catch on fire, DO NOT RUN. Walk purposefully
toward the fire shower station or the nearest fire blanket. Running will fan the
flames and intensify them.
3. Organic Solvents: Their Hazards
Avoid Contact with Organic Solvents. It is essential to remember that
most organic solvents are flammable and will burn if they are exposed to an
open flame or a match. Remember also that upon repeated or excessive
exposure some may be toxic or carcinogenic (cancer causing) or both. For
example, many chlorocarbon solvents, when accumulated in the body, cause
liver deterioration similar to the cirrhosis caused by the excessive use of
ethanol. The body does not rid itself easily of chlorocarbons nor does it
detoxify them; thus, they build up over time and may cause illness in the
future. Some chlorocarbons are also suspected to be carcinogens.
MINIMIZE YOUR EXPOSURE. Long-term exposure to benzene may
cause a form of leukemia. Don't sniff benzene, and avoid spilling it on
yourself. Many other solvents, such as chloroform and ether, are good
anesthetics and will put you to sleep if you breathe too much of them. They
subsequently they enhance its effect. Pyridine causes temporary
impotence. In other words organic solvents are just as dangerous as
corrosive chemicals, such as sulfuric acid, but manifest their
hazardous nature in other, more subtle ways.
If you are pregnant, you may want to consider taking this
course at a later time. Some exposure to organic fumes is inevitable,
and any possible risk to an unborn baby should he avoided.
In this textbook, carcinogenic chemicals or highly toxic solvents
are called for only in a few experiments and when they are
absolutely necessary to perform a particular procedure. Special
precautions will be given when the use of these chemicals is
required.
Minimize any direct exposure to solvents and treat them with
respect. The laboratory room should be well ventilated. Normal
cautious handling of solvents should not cause any health problem.

If you are trying to evaporate a solution in an open container, you

must do the evaporation in the hood. Excess solvents should he
discarded in a container specifically intended for waste solvents,
rather than down the drain at the laboratory bench.
A sensible precaution is to wear gloves when working with
solvents. Do Not Breathe Solvent Vapors. If you want to check the
odor of a substance (not a solvent), you should be careful, not to
inhale very much of the material. A technique for smelling minute
amounts of a substance is used. You should pass a stopper or spatula
moistened with the substance (if it is a liquid) tinder your nose.
Alternatively, you may hold the substance away from you and waft
the vapors toward you with your hand. But you should never hold
your nose over the container and inhale deeply:
On page 6 the hazards associated with organic solvents you are
likely to encounter in the organic laboratory are discussed in detail.
By using proper safety precautions, your exposure to harmful
organic vapors will be minimized and should present no health risk.
4. Waste Disposal
Do Not Place Any Organic Liquids or Solids into Sinks; Use
Waste Containers. Many organic substances are toxic. Flammable,
and difficult to degrade: it is not acceptable to dispose of organic
solvents or solids by pouring them down the sink. Municipal
sewage-treatment plants are not equipped to remove these materials
from sewage. Furthermore, with volatile and flammable materials, a
spark or an open flame can cause an explosion in the sink or further
down the drains.
The appropriate disposal method for wastes is to place them into
appropriately labeled waste containers. These containers should he
placed in the hoods in the laboratory.
Specific guidelines for disposing of waste will be determined by
the people in choice of your particular laboratory and by local
regulations. One system for handling waste disposal is presented
here. For each experiment in this textbook, you will he instru cted to
dispose of all wastes in one of the following ways:
Nonhazardous Solids: Nonhazardous solids such as paper and corks
can be placed into an ordinary wastebasket.
Broken Glassware: Broken glassware should be put into ordinary
wastebasket.
Organic Solids: Solid products that are not turned in or any other
organic solids should be disposed of in the container designated for

organic solids.
Inorganic Solids: Solids such as alumina and silica gel should he
placed into a container specifically designated for them.
Nonhalogenated Organic Solvents: Organic solvents such as diethylether, hexane, toluene, or any solvent that does not contain a halogen
atom, should he disposed of in the container designated for
nonhalogenated organic solvents.
Halogenated Solvents: Methylene chloride (dichloromethane),
chloroform, and carbontetrachloride are examples of common
halogenated organic solvents. Dispose of all halogenated solvents
into the container designated for them.
Strong Inorganic Acids and Bases: Strong acids such as
hydrochloric, sulfuric, and nitric acid and strong bases such as
sodium hydroxide and potassium hydroxide should be neutralized,
diluted with water, and poured down the drain.
Heavy Metals. Many heavy metal ions such as mercury and
chromium are highly toxic and should be disposed of into
specifically designated waste containers.
5. Use of Flames
Even though organic solvents are frequently flammable (for
example, hexane, diethyI-ether. methanol, acetone, and petroleum
ether), there are certain laboratory procedures for which a flame may
be used. Most often these procedures involve an aqueous solution. In
fact, as a general rule, use a flame to heat only aqueous solutions.
Heating methods that do not use a flame are discussed in detail . Most
organic solvents boil below 100C and an aluminum block, sand bath,
or water bath may be used to heat these solvents safely. A listing of
common organic solvents is given. Solvents marked in that table with
boldface type will burn. Diethyl ether, pentane, and hexane are
especially dangerous, because in combination with the correct
amount of air, they may explode.
Some common sense rules apply to using a flame in the
presence of flammable solvents. Again, we stress that you should
check to see whether anyone in your vicinity is using flammable
solvents before you ignite any open flame. If someone is using a
flammable solvent, move to a safer location before you light your
flame. Your laboratory should have an area set aside for using a
burner to prepare micropipets or other pieces of glassware.
The drainage troughs or sinks should never be used to dispose of
flammable organic solvents. They will vaporize if they are low boiling

and may encounter a flame further down the bench on their way to the
sink.
6. Inadvertently Mixed Chemicals
To avoid unnecessary hazards of tire and explosion, never pour
any reagent back into a stock bottle. There is always the chance that
you may accidentally pour back some foreign substance that will react
explosively with the chemical in the stock bottle. Of course, by
pouring reagents back into stock bottles you may introduce impurities
that could spoil the experiment for the person using the stock reagent
after you. Pouring things back into bottles is thus not only a
dangerous practice, but it is also inconsiderate. This also means that
you should not take more chemicals than you need.
7. Unauthorized Experiments
You should never undertake any unauthorized experiments. The
risk of an accident is high, particularly with an experiment that has not
been completely checked to reduce the hazard. You should never work
alone in the laboratory. The laboratory instructor or supervisor must
always be present.
8. Food in the Laboratory
Because all chemicals are potentially toxic, you should avoid
accidentally ingesting any toxic substance; therefore, never eat or
drink any food in the laboratory. There is always the possibility that
whatever you are eating or drinking may become contaminated with a
potentially hazardous material.
9. Clothing
You should always wear shoes in the laboratory. Even open -toed
shoes or sandals offer inadequate protection against spilled chemicals
or broken glass. Do not wear your best clothing in the laboratory
because some chemicals can make holes or permanent stains on your
clothing. To protect yourself and your clothing, it is advisable to wear
a full-length laboratory apron or coat. When working with chemicals
that are very toxic, you should wear some type of gloves. Polyethylene
gloves provide good protection. Disposable surgical gloves may offer
protection when working with some chemicals. On a final note, you
should tie back hair that is shoulder length or longer, especially if you
are working with a burner.

10.

First Aid: Cuts, Minor Burns, and Acid or Base Burns

Note: Any injury, no matter how small, must be reported to your

instructor immediately.
If any chemical enters your eyes, immediately irrigate the eyes
with copious quantities of water. Tempered (slightly warm) water if it
is available, is preferable. Be sure that the eyelids are kept open.
Continue flushing the eyes in this way for 15 minutes.
In case of a cut, wash the wound well with water, unless you
are specifically instructed to do otherwise. If necessary, apply
pressure to the wound to stop the flow of blood.
Minor burns caused by flames or contact with hot objects may be
soothed by immediately immersing the burned area in cold water or
cracked ice for about 5 minutes. Applying salves to burns is
discouraged. Severe burns must be examined and treated by a
physician.
For chemical acid or base burns, rinse the burned area with
copious quantities of water for at least 10 minutes. Then you can
neutralize this area use 5% solution of NaHCO 3 (in the case of acid)
or 0,5% solution of acetic acid (in the case of base) to wash skin!
If you accidentally ingest a chemical, immediately begin
drinking large quantities of water while proceeding immediately to
the nearest medical assistance. It is important that the examining
physician he informed of the exact nature of the substance ingested.
CO MMO N S O L VE NT S
Most organic experiments involve an organic solvent at some
step in the procedure. A list of common organic solvents follows,
with a discussion of toxicity, possible carcinogenic properties, and
precautions that you should use when handling these solvents. A
tabulation of the compounds currently suspected of being
carcinogens appears at the end of this chapter.
Acetic Acid. Glacial acetic acid is corrosive enough to cause
serious acid burns on the skin. Its vapors can irritate the eyes and
nasal passages. Care should be exercised not to breathe the vapors
and not to allow them to escape into the laboratory.
Acetone. Relative to other organic solvents, acetone is not very
toxic. It is flammable, however. Do not use near open flames.
Benzene. Benzene can cause damage to bone marrow; it is a

cause of various blood disorders, and its effects may lead to

leukemia. Benzene is considered a serious carcinogenic hazard.
Benzene is absorbed rapidly through the skin. It also poisons the
liver and kidneys. In addition, benzene is flammable. Because of its
toxicity and its carcinogenic properties, benzene should not be used
in the laboratory; you should use some less dangerous solvent
instead. Toluene is considered a safe alternative solvent in
procedures that specify benzene.
Carbon Tetrachloride. Carbon tetrachloride can cause serious
liver and kidney damage as well as skin irritation and other
problems. It is absorbed rapidly through the skin. In high
concentrations, it can cause death, owing to respiratory failure.
Moreover, carbon tetrachloride is suspected of being a carcinogenic
material. Although this solvent has the advantage of being
nonflammable (in the past, it was used on occasion as a fire
extinguisher), it should not be used routinely in the laboratory since
it causes health problems. If no reasonable substitute exists,
however, it must be used in small quantities, as in preparing
samples for infrared (IR) and nuclear magnetic resonance (NMR)
spectroscopy. In such cases, you must use it in a hood.
Chloroform. Chloroform is like carbon tetrachloride in its
toxicity. It has been used as an anesthetic. However, chloroform is
currently on the list of suspect carcinogens. Because of this, do not
use chloroform routinely as a solvent in the laboratory.
Occasionally, it may be necessary to use chloroform as a solvent
for special samples. Then, you must use it in a hood. Methylene
chloride is usually found to be a safer substitute in procedures that
specify chloroform as a solvent. Deuterochloroform CDCI3 is a
common solvent for NMR spectroscopy. Caution dictates that you
should treat it with the same respect as chloroform.
1,2-Dimethoxyethane (Ethylene Glycol Dimethyl Ether or
Monoglyme). Because it is miscible with water, it is a useful
alternative to solvents such as dioxane and tetrahydrofuran, which
may be more hazardous. 1,2-Dimethoxyethane is flammable and
should not be handled near open flames. On long exposure to light and
oxygen, explosive peroxides may form. 1,2-Dimethoxyethane is a
possible reproductive toxin.
Dioxane. Dioxane has been used widely because it is a
convenient, water-miscible solvent. It is now suspected, however, of
being carcinogenic. Additionally, it is toxic, affecting the central
nervous system, liver, kidneys, skin, lungs, and mucous membranes.
Dioxane is also flammable and tends to form explosive peroxides

when it is exposed to light and air. Because of its carcinogenic

properties, it is no longer used in the laboratory unless absolutely
necessary. Either 1.2-dimethoxyethane or tetrahydrofuran is a suitable,
water-miscible alternative solvent.
Ethanol. Ethanol has well-known properties as an intoxicant. In
the laboratory, the principal danger arises from fires, since ethanol is
a flammable solvent. When using ethanol, take care to work where
there are no open flames.
Diethyl Ether (Ether). The principal hazard associated with
diethyl ether is fire or explosion. Ether is probably the most
flammable solvent one is likely to find in the laboratory. Because the
vapors are much more dense than air, they may travel along a
laboratory bench for a considerable distance from their source before
being ignited. Before you begin to use ether, it is very important to be
sure that no one is working with matches or any open flame. Ether is
not a particularly toxic solvent, although in high enough
concentrations it can cause drowsiness and perhaps nausea. It has
been used as a general anesthetic. Ether can form highly explosive
peroxides when exposed to air. Consequently, you should never distill
it to dryness.
Hexane. Hexane may be irritating to the respiratory tract. It can
also act as an intoxicant and a depressant of the central nervous
system. It can cause skin irritation because it is an excellent solvent
for skin oils. The most serious hazard, however, comes from its
flammable nature. The precautions recommended fo r the use of
diethyl ether in the presence of open flames apply equally to hexane.
Methanol. Much of the material outlining the hazards of ethanol
applies to methanol. Methanol is more toxic than ethanol; ingestion
can cause blindness and even death. Because methanol is more
volatile, the danger of fires is more acute.
Methylene Chloride (Dichloromethane). Methylene chloride is
not flammable. Unlike other members of the class of chlorocarbons, it
is not currently considered a serious carcinogenic hazard. Re cently,
however, it has been the subject of much serious investigation, and
there have been proposals to regulate it in industrial situations where
workers have high levels of exposure on a day-to-day basis.
Methylene chloride is less toxic than chloroform and carbon
tetrachloride. It can cause liver damage when ingested, however, and
its vapors may cause drowsiness or nausea.
Pyridine. There is some fire hazard associated with pyridine.
The most serious hazard arises from its toxicity, however. Pyridine
may cause depression of the central nervous system; irritation of the

skin and respiratory tract; damage to the liver, kidneys, and

gastrointestinal system; and even temporary sterility. You should
treat pyridine as a highly toxic solvent and handle it only in the fume
hood.
Tetrahydrofuran. Tetrahydrofuran may cause irritation of the
skin, eyes, and respiratory tract. It should never be distilled to
dryness, since it tends to form potentially explosive peroxides on
exposure to air. Tetrahydrofuran does present a fire hazard.
Toluene. Unlike benzene, toluene is not considered a
carcinogen. However, it is at least as toxic as benzene. It can act as
an anesthetic and cause damage to the central nervous system. If
benzene is present as an impurity in toluene, then one must expect
the hazards associated with benzene to manifest themselves. Toluene
is also a flammable solvent, and the usual precautions about working
near open flames should he applied.
You should not use certain solvents in the laboratory because of
their carcinogenic properties. Benzene, carbon tetrachloride,
chloroform, and dioxane are among these solvents. For certain
applications, however, notably as solvents for infrared or NMR
spectroscopy, there may be no suitable alternative solvent. When it
is necessary to use one of these solvents, safety precautions are
recommended.
Because relatively large amounts of solvents may be used in a
large organic laboratory class, your laboratory supervisor must take
care to store these substances safely. Only the amount of solvent that
is needed for a particular experiment being conducted should be kept
in the laboratory room. The preferred location for bottles of solvents
being used during a class period is in a hood. When the solvents are
not being used, they should be stored in a fireproof storage cabinet
for solvents. If possible, this cabinet should be ventilated into the
fume hood system.

10

II. Laboratory Techniques

II.1. Proper Use of Heating Equipment
Introduction
A variety of methods for heating chemicals can be used in the
laboratory. The ones we shall include are the Bunsen burner, the hot plate,
the heating mantle, the oil bath.
Heating with a Bunsen Burner
Discussion
The Bunsen burner is used routinely in the general chemistry
laboratory, but they are to be avoided if possible in the organic laboratory.
Many organic chemicals are flammable, and an open flame is an invitation to
starting a laboratory fire or an explosion of combustible vapors.
WARNING! Never use a bunsen burner in the presence of diethyl ether or
other highly flammable solvents.
In the organic chemistry laboratory, the Bunsen burner provides a
high-temperature heat source (1000C) for heating water or for heating a
water-solid suspension. The Bunsen burner can also be used for the rare
distillation in which a high temperature is required.
Apparatus
Figure 1. shows the proper method of using the Bunsen burner for
heating.

11

Figure 1. Heating a beaker with a Bunsen burner.

Procedure
When heating a glass flask of any type, insert either a wire screen, or a
wire screen-asbestos pad between the flame and the flask (shown in Figure
1 ) to distribute the heat evenly. Distributing the heat is necessary to prevent
cracking of flasks, especially the round-bottomed type.
Heating with a Hot Plate
Discussion
The variable-temperature hot plate is a very versatile heating unit. If
available, it can be used under almost any circumstances, although it is not
practical for heating round-bottomed flasks. The main advantage of using a
hot plate is that it provides a flameless variable-temperature heat from a flat
surface, so no additional support is normally required when using a flatbottomed flask.
WARNING! The hot surface can ignite some flammable substances
especially ether that have low auto ignition temperatures.
Apparatus

12

A hot plate with a variable-temperature control and no exposed heating

coils is readily available in most chemistry laboratories.
Procedure
For liquids with low boiling points, use the lower temperature settings
of the hot plate. For liquids with higher boiling points, use the higher
temperature settings.
CAUTION: Place applicator sticks or boiling chips in the flask to prevent
bumping.
Heating with a Heating Mantle
Discussion
A heating mantle is a specialized kind of heating device designed to be
used only with round-bottomed flasks when liquids are being heated under
reflux or are being distilled. There are different heating mantle is required for
each size of flask.

Figure 2. Variac and a cutaway view of a heating mantle containing a

round-bottomed flask.

Apparatus
Figure 2. shows a Variac and heating mantle that fits flasks ranging in
size from 25 mL to 500 mL.

13

Procedure
The heating temperature at which a heating mantle operates can
be controlled by plugging it into a Variac, as shown in Figure 2. A low
voltage setting on the Variac allows one to operate the heating mantle at
low temperatures. As the voltage setting is increased, the temperature at
which the heating mantle operates also increases. When electric power
is being received by the heating coil, most models display a red
indicator light. Start the Variac at a low voltage setting and increase
gradually until you get the proper boiling rate.
Heating with an Oil Bath
Discussion
An oil bath serves exactly the same purpose as a heating mantle
and is used in exactly the same way. An oil bath consists of a pan of
mineral oil containing a heating coil in the bottom that is plugged into a
Variac. Varying the voltage controls the temperature of the oil. A low
voltage gives a low temperature. As the voltage is increased, the
temperature of the oil bath increases. An oil bath should be used only to
heat round-bottomed flasks under reflux or during a distillation.
Apparatus
Figure 3. shows a proper oil-bath apparatus.

14

Figure 3. Hearing a round-bottomed flask with an oil bath.

Procedure
When you use an oil bath, you must support the round-bottomed
flask by clamping it to a ring stand. As shown in Figure3., only half
the flask should be immersed in the oil. Start the Variac at a low
voltage setting and increase gradually until you get the proper reflux or
distillation rate. W A R N I N G ! Hot oil is dangerous and can cause
severe burns. It also has a bad odor and makes flasks dangerously
slippery. Do not use an oil bath if a heating mantle is available.
WARNING! Accidental addition of water to a hot oil bath can cause
serious splattering.
Heating with a Hot-water Bath
Discussion
The hot-water bath consists of just a beaker or a pan of boiling water.
Apparatus and Procedure

15

To make the hot-water bath, heat a beaker of water to boiling, using a

hot plate or if no flammables are involved, use a Bunsen burner. CAUTION:
Be sure to put boiling chips in the water to prevent bumping.
Heating Under Reflux
Discussion
When a liquid boils, it vaporizes and thus can escape from a vessel.
The amount of liquid solvent in the flask is thereby reduced. If the solvent
vapors are cooled in a condenser to regenerate the liquid state, the liquid
returns to the flask and no solvent is lost. This process is called heating
under reflux or refluxing.
Apparatus
CAUTION: Before you insert any ground-glass joints into the
flask, cover them with a very thin layer of stopcock grease.

16

Figure 4. Apparatus for heating a liquid under reflux. Ground-glass joints

should be greased very lightly by applying stopcock grease to the glass joint
and then removing the excess with a paper towel.
After applying the grease to the glass joint, wipe clean with a paper
towel to remove the excess. CAUTION: Any excess grease on a ground-glass
joint goes into the flask when heated.
The grease prevents the two ground-glass joints from fusing. This is
especially necessary when dealing with alkaline solutions. WARNING! Do
not stopper the top of the reflux system. If a reflux system is not open to
the atmosphere, it will explode.
Procedure
17

Place the liquid to be refluxed in the flask and add three or four
boiling chips. Bring the liquid to a gentle boil, using the appropriate heating
apparatus. The vaporized solvent will condense in the condenser and fall
back into the reaction flask. The condenser must have cold water flowing
through it as shown.
CAUTION: Three to four boiling chips are required to prevent
bumping. Bumping is a process in which a solvent becomes superheated and
then undergoes a sudden release of a large vapor bubble, explosively forcing
liquid out of the flask.
CAUTION: When you are heating a suspension of a solid in a liquid
under reflux, the liquid must be boiled gently to prevent collection of the solid
material in the condenser. Otherwise the condenser may become plugged,
creating an explosion in the flask as the pressure of the evaporated gases
increases.
II.2. Drying Liquids.
Introduction
Water is slightly soluble in most organic liquids. Water introduced
during a reaction or extraction procedure can interfere with subsequent
reactions, crystallization of a product, determination of a melting point, or
obtaining an infrared spectrum. Thus water is removed from an organic liquid
by adding a drying agent.
Low-boiling-point organic solvents, or on occasion a high-boilingpoint liquid, are removed from a dissolved solid by evaporation. Solvents are
removed under reduced pressure if there is a possibility of decomposition of
components at medium to low temperatures.
Drying Organic Liquids
Discussion
Several anhydrous inorganic salts that react with water to form
hydrated salts are useful as drying agents for removing water from organic
solvents.
MgSO4 + 7 H20 MgSO4x7xH2O
The various drying agents differ in effectiveness, rate of removal,
and capacity to remove water from an organic liquid (Table 1.). Their
effectiveness in removing all water present is the major criterion for
choosing which drying agent to use. However, the chemical properties of
the drying agents shown in table limit their use. Magnesium sulfate, a

18

Lewis acid, is not generally used with amines, and sodium or

potassium carbonate cannot be used with acids.
Procedure
Add a volume of powdered drying agent equal to
approximately 5% of the volume of the liquid to be dried. Gently swirl
the liquid-drying agent mixture for 4 to 5 minutes or longer as directed.
The drying agent tends to become gummy as it reacts with water and
sticks to the sides of the vessel. If all the drying agent appears gummy
or if you are in doubt that all possible drying has taken place add an
additional 5% by volume of the drying agent. After drying is
complete, remove drying agent by gravity filtration, using a fineporosity filter paper.
Name
Magnesiu
m supfate

Hydrate

Effectivene
ss

Rate

Capacit
y

Use

MgSO4x7H2
O

Medium

Rapid

High

General
except for
amines

Sodium
sulfate
Calcium
chloride

Na2SO4x7H2
O

Low

Mediu
m

High

General

CaCl2x2H2O

High

Rapid

Low

Hydrocarbon
es, alkyl or
aryl halides

Calcium
sulfate
Sodium
carbonate

CaSO4x1/2H2
O

High

Rapid

Low

General

Potassium
carbonate

Molecule
sieves

Na2CO3x10H
2O

Medium

Mediu
m

Mediu
m

K2CO3x1/2H2
O

Medium

Mediu
m

Mediu
m

High

High

High

Table 1. Drying agents

II. 3. Evaporation of a Liquid

19

Amines,
esters,
amides,
ketones,
alcohols
Amines,
esters,
amides,
ketones,
alcohols
General

Discussion
To recover a solid dissolved in a liquid, you can evaporate the
solution to dryness. You can also remove low-boiling-point liquids
from high-boiling-point liquids before you proceed with an
experiment.
The Rotary E v a p o r a t o r
This piece of apparatus, usually communal in the teaching laboratory, is
designed for the rapid removal of large quantities of volatile solvent at
reduced pressure from solutions, leaving behind a relatively involatile
component.

Figure 5. Typical example of a rotary evaporator (the clip holding the

evaporating flask has been omitted for clarity).
Rotary evaporation finds greatest use for removing extraction and
chromatography solvents used in the isolation and purification of reaction
products. The principle of operation which distinguishes the apparatus from
that used for ordinary reduced pressure distillation is the fact that the
distillation flask is rotated during the removal of solvent.
A wide range of designs is available commercially for a variety of
specific uses, but a basic model most commonly encountered in the
teaching laboratory is depicted in Figure 5.
Correct Use of the Rotary Evaporator
Ensure that the receiving flask is empty of solvent and that water is
passing through the condenser coils at a slow but steady rate. Turn on the

20

water aspirator to its fullest extent and then attach the evaporating flask (with
adapter if required) to the vapor duct, using a clip to ensure that the flask
stays in place. Support the flask lightly with the hand, commence slow
rotation and then close the stopcock at the end of the condenser. When the
manometer indicates a significant reduction of pressure within the system (if
no manometer is attached to the apparatus, listen for a marked change in tone
of the sound made by the water rushing from the aspirator) it is safe to
remove your hand and regulate the speed of rotation to spread the solvent out
around the flask without causing it to splash. If the mixture commences to
boil uncontrollably, temporarily open the stopcock at the top of the condenser
to allow entry of air and then reclose the stopcock. Once the evaporation
from the solution has stabilized, the evaporating flask may be introduced into
a bath of warm water if desired. However, be ready to remove the flask
immediately there is any indication that the mixture is beginning to boil too
vigorously. The majority of common solvents such as ether or light
petroleum have boiling points well below room temperature at the reduced
pressures possible using this system, so you must exercise great care when
heating the evaporating flask. With the more volatile solvents it is advisable
to place the flask in a cold water bath at the outset and then allow the bath to
warm up slowly during the course of solvent removal. The last traces of
solvent are difficult to remove from samples, particularly the kinds of gummy
materials which are often isolated from reactions, and so it is necessary to
leave the residue on the rotary evaporator for at least 5 min after the last of
any solvent has been seen running into the receiver.
When you are satisfied that all of the solvent has been removed
from your sample, stop the flask rotating and raise it from the heating
bath. If you reverse the order of these operations you will get wet with
spray from the flask! Open the stopcock to allow air into the system,
supporting the Ink with your hand, remove the flask and turn off the
aspirator and condenser water. Empty the contents of the receiver
flask into the container designated for used solvents (not down the
sinks!) and check that you have left no material adhering to the inside
of the vapor duct as this will not only reduce your yield, but also
contaminate the next user's sample.
Vacuum Pumps
The common procedures which call for reduced pressure in the
organic chemistry laboratory are filtration with suction and reduced
pressure distillation. The former technique, which simply requires a
source of suction, is adequately served by use of a water aspirator,
although the reduced pressures which can he achieved with this

21

simple apparatus (ca. 1020 mmHg) are frequently sufficient for

use with reduced pressure distillations. However, very high boiling
materials, or purifications involving sublimations require recourse
to a vacuum of 0.1-1.0 mmHg which is provided by an oil
immersion rotary vacuum pump. A higher vacuum is achieved by
using a mercury vapor diffusion pump in series with a double action
rotary oil pump, but such conditions are required only rarely in the
laboratory and will not be discussed here. It should not be forgotten,
however, that mass spectrometric analysis would be impossible
without the ability to achieve reduced pressures in the region of 10 -6
mmHg.
Using a Water Aspirator
Whereas the flow of water passing through a condenser does not
need to be more than a gentle trickle, water aspirators must never be
used with the water at less than full blast. (Why do students always
seem to get this one the wrong way round?) With the water turned
on full, check that the air bleed screw on the side arm is open (as
these little screws have a tendency to become stuck or get lost, some
aspirators do not have them, but an additional stopcock attached to
the water trap will serve the same purpose) and then attach the
pressure tubing to the apparatus to be evacuated. Close the air bleed
(or stopcock on the water trap) and observe the pressure drop on the
manometer. For vacuum filtrations, the actual quality of the vacuum
is largely unimportant but, of course, it will be necessary to note and
regulate this pressure if you are carrying out a vacuum distillation.
The critical stage in working with water aspirators comes when
the vacuum is to be released. It is imperative that the water supply to
the aspirator remains on until the pressure within the system has been
allowed to return to that of the atmosphere. If you do not observe
this simple procedure, the inevitable result will be a suck -back of
water into the apparatus.
With suction filtration it is frequently sufficient simply to remove the
pressure tubing from the side arm of the receiving vessel before
turning off the tap, although this lazy practice runs the risk of spillage
when the tubing is suddenly removed and air rushes into the
receiver. The correct procedure for both reduced pressure filtration
and distillation involves the unscrewing of the air bleed on the side
arm of the aspirator (or opening the stopcock on the water trap) until
the manometer registers a steady increase in pressure within the
system, or the lone of the water rushing through the aspirator changes

22

abruptly. Do not continue to unscrew the air bleed, otherwise the screw
will drop out and be lost for ever down the sink. It is always good
practice when carrying out reduced pressure distillations to allow the
residue in the distilling flask to cool down to near room temperature before
admitting air, particularly if the flask has been heated strongly during
distillation. The
II.4. Stirring
There are three main ways that mixtures can be agitated by
hand, with a magnetic stirrer and with a mechanical stirrer; only the
last two require any particularly sophisticated equipment! Remember
that homogeneous solutions do not in general require any stirring
after the initial mixing. The exceptions to this rule are reactions
which are carried out at low temperatures (for instance, reactions
involving alkyl lithium reagents or diisobutylaluminum hydride),
and in such cases, agitation . is required for heat dispersal rather than
for mixing of reagents.
Magnetic Stirrers
Magnetic stirring is the method of choice if an extended period
of continuous agitation is required, since it is easy to set up the
apparatus; particularly for small scale set-ups or closed systems. The
main drawback to the technique is that it cannot cope with viscous
solutions or reactions which contain a lot of suspended solid. In
addition, volumes of liquid much greater than I L are not stirred
efficiently throughout their whole bulk. The magnetic stirrer may also
be equipped with a hotplate, and these combined stirrer hotplates are
particularly versatile pieces of apparatus. In general, the larger the
volume of material to be stirred, the more powerful the motor needed
and the longer the magnetic stirrer bar required.
Mechanical Stirrers
Larger scale reactions or viscous mixtures require the greater power
of an external motor unit turning a stirrer blade. It is highly advantageous
for the motor to possess a variable speed control and a typical model is
shown in Figure 6. These units are rather heavy and so it is necessary to
support them firmly.

23

Figure 6. Typical mechanical stirrer.

II. 5. Separation of Compounds by Distillation Introduction
All distillations depend on the difference between the boiling points of
the compounds being separated. As the temperature of a solution rises, the
substance that has the lower boiling point vaporizes faster than the substance
that has the higher boiling point. Thus the vapor formed is richer in the
lower-boiling-point substance than the original solution was. However, some
liquid mixtures form an azeotrope, or constant boiling-point mixture. In
such cases, the vapor formed by boiling the solution has the same
composition as the liquid azeotrope.
Simple Destillation
Discussion
Knowing how to use simple distillation is of great value in separating
a liquid from a solid, or in separating two liquids that have a large difference
between their boiling points. When you are separating a liquid from a solid, a
simple distillation apparatus (Figure 6.) is sufficient to separate the
components completely.

24

Figure 6. Apparatus for simple distillation. Each piece that could possibly
fall should be attached with a clamp to a ring stand. Ground-glass joints
should be greased very slightly by applying stopcock grease and then
wiping with a paper towel.
For two liquids with widely different boiling points, a good but not a
total separation will be obtained. If the liquids have similar boiling points, the
first fraction collected will contain a greater proportion of the low-boilingpoint liquid than was present in the original solution. The separation of
liquids with similar boiling points can thus be considered only a crude one.
Apparatus
Figure 6. shows the proper setup for a simple distillation apparatus.
All ground-glass joints should be lightly greased to prevent fusing. This is
done by applying stopcock grease to the glass joint and then wiping with a
clean paper towel.
The vacuum adapter outlet must be open, so that distillation is done at
atmospheric pressure and so that no pressure builds up as vapors form.
Procedure
Use a heating mantle or a steam cone as the heat source for most
distillations. Place the solution or mixture to be distilled in the roundbottomed flask attached to the distilling head. Add a few boiling chips or an

25

applicator stick. Be sure that cold water is flowing through the condenser in
the proper direction (bottom to top). Turn on the heating device and adjust it
to an appropriate setting to obtain the temperature necessary for a moderate,
easily controlled rate of distillation.
CAUTION: Do not overheat. Overheating causes too-rapid distillation. This
results in poor separation of the components.
Collecting Fractions of Distilled Liquid
There are two basic methods for collecting fractions of distilled
liquids. In the simpler method, you must decide before beginning the process
what distillation temperature increments are to be used in changing the
vessels in which the distillation is being collected. We might, for instance,
collect all the liquid distilling between 20C and 60C in the first flask and
label it "Fraction 1." The second fraction might contain the liquid collected in
the 6080C range, and so on. When we collect fractions by this method,
we usually discard all fractions but the one we intend to subject to further
distillation. If we are distilling the liquid from a solution containing a solid,
we simply collect the liquid in one fraction and either use it or discard it as
directed.
In the second method of collecting fractions, you remove the initial
collection vessel as soon as you obtain a relatively constant distillation
temperature. The liquid obtained at the relatively constant temperature is the
second fraction. As soon as the temperature begins to undergo a definite rise,
you again replace the collection flask. This process is continued until all the
liquid has been distilled. Figure 7. is a graph showing a hypothetical
separation of two liquids into several fractions.

26

Figure 7.: Idealized distillation curve for a solution composed of two miscible
(soluble) liquids.
Fractional Distillation
Discussion
Fractional distillation is designed to separate liquids that have only a
small difference between their boiling points. The fractional distilling
columns shown in Figure 8. are the essential instruments for separating
liquids with similar boiling points. Although there is a variety of fractional
distillation columns, only three are in common use in undergraduate organic
chemistry. The Vigreux column, which is effective only for total separation of
liquids having a difference in boiling points in excess of 60C.
The other two fractional distillation columns in Figure 4.7. are just 20
cm condensers packed with glass beads, glass helices, little glass rings
(Raschig rings), the commercial product Heli-Pak, or stainless-steel wool
sponge. A packing of glass helices in a 20 cm condenser makes a fractional
distillation column capable of completely separating liquids differing in
boiling points by 36C. The Heli-Pak makes the more effective fractional
distillation column. A column of 20 cm of Heli-Pak allows complete
separation of liquids with a difference in boiling points of only I7C.

27

Figure 8.: Three fractional distillation columns: (a) Vigreux column;-(b)

column packed with glass beads; (c) column packed with stainless steel wool
sponge.
Stainless-steel wool sponge, one of the cheapest packings. Steel wool
has the disadvantage of reacting with some organic chemicals, such as alkyl
halides. Of course, some liquid is lost in the distillation process by hold-up in
the fractional distillation column.
Apparatus
Figure 9. shows the proper setup of a fractional distillation apparatus.
All ground-glass joints should be lightly greased by applying stopcock grease
to the glass joint. They should then be wiped with a clean paper towel.
Procedure
Distill the liquid slowly, preferably using a heating mantle to enhance
separation of components. If the laboratory temperature is low, the fractional
distillation column may have to be insulated by wrapping it with a narrow
strip of aluminum folded over a thin layer of glass wool to form a flat strip.
Insulation of fractional distillation columns maintains a proper temperature
gradient, allowing one to collect 510 drops of distillate per minute. Change
the collection flask each time you have a significant temperature change.

28

Figure 9.: Apparatus for fractional distillation. Each piece that could
possibly fall should be attached with a clamp to a ring stand. Ground-glass
joints should be greased very lightly by applying stopcock grease and then
wiping with a paper towel.
The first flask will contain only the liquid distilled before the
temperature is stabilized. The second flask will contain the liquid of lowest
boiling point after the temperature stabilizes. Once the temperature begins
to increase by 2-VC, use another collection flask. As soon as the distillation
temperature again becomes constant, use still another collection flask. This
procedure continues until all the liquid has been distilled. Figure 7. is a
graph of temperature versus time. It shows the temperature ranges at which
various fractions are collected.
Vacuum Distillation
Introduction

29

Many liquids with high boiling points may decompose if distilled at

atmospheric pressure. To prevent a liquid from decomposing, we distill it
under reduced pressure, a technique called vacuum distillation. As the
atmospheric pressure in the distillation flask is reduced, the boiling point is
lowered, since a liquid boils when its vapor pressure is equal to the pressure
in the distillation flask. Thus, if the pressure in the flask is reduced to near
vacuum conditions, the liquid boils at a temperature far lower than its 1atm-pressure boiling point. To successfully conduct a vacuum distillation,
one must consider several factors. We shall describe each factor in detail in
separate sections.
Figure 10. illustrates the relationship between boiling point and pressure.
Note the large decrease in boiling point as the pressure of a system
approaches 0 Ton. Thus to effectively reduce a compound's boiling point the
pressure must be reduced to less than 50 Torr.

Figure 10.: Boiling point versus pressure for water, benzaldehyde and
diethylaniline.
To understand the use of manometers, remember that standard
atmospheric pressure is 760 Torr, which is a pressure capable of supporting a
760-mm column of mercury in an evacuated glass tube. This system is called
a Torricelli barometer (Figure 5.2.). Therefore a pressure of 1.0 Torr is equal
to the pressure exerted by a 1.0-mm column of mercury.
Apparatus
If the organic kit does not contain an ebulliator, construct one from a
6-mm glass or disposable pipet. Two styles of ebulliator (Figure 11.) can be
30

made. Style (a) has a larger capillary tube and is stiff. The last 5-10 cm of style
(b) is a very thin capillary tube which is flexible enough to stir the solution as
air bubbles pass through it during the vacuum distillation. Figure 12.
illustrates a properly assembled apparatus for vacuum distillation.
CAUTION: Lightly grease all ground-glass joints with Lubriseal, Apiezon, or
other stopcock grease before use. All rubber tubing except for the two tubes
on the condenser must be heavy-walled pressure tubing.
The system is normally connected to a water aspirator. The trap is necessary to
pre-vent contamination of the system by backup of water from the aspirator if
there is a sudden change in water pressure in the line.

Figure 11.: Two styles of ebulliators: (a) stiff end; (b) flexible end.

31

Figure 12.: Vacuum distillation apparatus including ebulliator, manometer

and trap.
Each piece that could possibly fall should be attached with a clamp to
a ring stand. All ground-glass joints should be greased very lightly by
applying stopcock grease and then wiping with a paper towel.
Fraction Collectors
Several types of fraction collectors are available. One is shown in
Figure 13. This is a special vacuum adapter having four ground-glass joints to
which flasks are attached. To change collection flasks, rotate the apparatus
until another flask is in position.

32

Figure 13.: Fraction collectors for use in vacuum distillations.

Closed-end Manometer
Figure 14. shows a standard commercial model and a laboratory
model of a manometer with one closed end. As shown in part (b), the closed
arm of the manometer, when not connected to a vacuum distillation system, is
full of mercury. After the open arm of the manometer is connected to a vacuum
distillation system, the arm is partially evacuated. When the pressure is
sufficiently reduced, there is a shift of the mercury into the arm open to the
vacuum distillation system as shown in part (c).

33

Figure 14. Closed-end manometers: (a) commercial type; (b) laboratory type
not attached to vacuum distillation system; and (c) laboratory type attached to
vacuum distillation system.
The pressure above the mercury in the closed arm is 0.0 Ton.
Therefore, the difference in mercury levels between the two columns is a
direct measure of the partial pressure of the vacuum distillation system to
which it is connected. Remember that 1.0 mm of mercury is equal to a
pressure of 1.0 Ton. Psystem = Hg
Open-end Manometer
The least-used manometer is the open-end manometer, which is
simply a U-tube in which each side is 1 meter long. Part (a) of Figure 15.
shows that if the pressure is identical in both arms the mercury level is
identical in both arms. When a partial vacuum is created in one arm, there is a
corresponding shift in the mercury column into the arm that is under reduced
columns is a direct measure of the reduction of the pressure in the system
compared to the atmospheric pressure in the laboratory. The partial pressure
in the vacuum distillation system can be determined by use of the following
equation: Psystem = Patmosphere - Dhmanometer.

34

Figure 15.: Open-end manomether

Distillation Procedure
Step 1: Assemble the apparatus illustrated in Figure 12., including trap and
attached manometer.
Step 2: Attach system to water aspirator.
Step 3: Place liquid to be distilled in distillation flask.
Step 4: Turn on water to condenser.
Step 5: Turn water aspirator on by opening valve to maximum water flow.
Step 6: Raise heating mantle or oil bath and slowly raise temperature until
distillation begins.
CAUTION: Raising the temperature too rapidly can cause overheating and
decomposition of material, creating a fog in the distillation flask.
If decomposition begins due to overheating, remove source of heat and cool
before proceeding.
Step 7: Maintain a distillation rate of approximately one drop per second.
Changing Collection Flasks
If no fraction collector is available, single flasks must be used. To
change single flasks, follow this procedure when distillation temperature
begins to increase.
Step 1: Turn off heat source and lower it so heating of flask is discontinued.

35

Step 2: Turn off vacuum by removing of vacuum tube from vacuum adapter,
and wait for the pressure of system has reached equilibrium with atmospheric
pressure. Replace collection flask.
Step 3: Turn on vacuum.
Step 4: Raise heat source into position and turn on to continue distillation.
Shut-down Procedure
Step 1: Remove and turn off heat source.
Step 2: Turn off vacuum by removing of vacuum tube from the vacuum
adapter, and wait for the pressure of system has reached equilibrium with
atmospheric pressure.
Steam Distillation
Introduction
One can also use distillation to separate liquid mixtures in which the
liquids are not totally miscible or soluble. The procedure involves the use of
water, in the form of liquid or steam, to create a mixture of two immiscible
liquids in the distillation flask. Since the solvent mixture always boils at a
temperature below that of either component, high-boiling-point substances
can be removed from natural sources or tarry mixtures at low temperature.
This prevents possible decomposition. When the vapors are condensed, the
organic liquid usually separates from the water.
Live-steam Distillation
Discussion
Live-steam distillation is ideally suited for use with finely powdered
materials or unusually viscous liquid substances.
Apparatus
Figure 16. shows the proper setup of apparatus for live-steam
distillation. Note that a bent 67 mm glass tube extends to the bottom of the
three-necked round-bottomed flask. Grease ground-glass joints very lightly by
applying stopcock grease to a glass joint and then wiping with a clean paper
towel.

36

Figure 16.: Apparatus for live steam distillation. Each piece that could
possibly fall snout, be attached with a clamp to a ring stand. Ground-glass
joints should be greased very lightly by applying stopcock grease and wiping
with a paper towel.
Procedure
Set up the apparatus as shown in Figure 16.. No boiling chips are
necessary. Place the liquid or solid to be subjected to steam distillation in the
three-necked round-bottomed flask. If the material is a solid, add enough
water to just cover the solid. Open the screw clamp on the rubber tubing at the
bottom of the steam trap. Connect the top tube of the steam trap to the boiler
and turn on the steam. Leave the bottom tube on the steam trap open until all
the water drains away and a steady flow of steam is established. Then close
the steam trap. This causes the steam to flow into the distillation flask.
During the distillation, water collects in the steam trap and must be drained
out occasionally.
Collect the condensed water and organic material as a single fraction.
It is advisable to complete a steam distillation in one laboratory session, but if
you can't, use the following procedure.
II.6. Filtration
There are few experiments in organic chemistry that do not involve at
37

least one filtration step. Filtration of a suspension to remove insoluble solids

is a fundamental technique of preparative chemistry and is achieved by
allowing the liquid to pass through a porous barrier, such as filter paper or
sintered glass, whereby the solid remains on the barrier. In many cases,
gravity alone is sufficient force for the liquid to pass through the porous
barrier; this is referred to as gravity filtration. However, many organic
solids are bulky, and filter slowly under gravity alone. In these cases the
process is speeded up considerably by using the suction or vacuum
filtration technique, in which a partial vacuum is applied to the filter flask
(which must be thick-walled), and hence air pressure on the surface of the
liquid in the filter funnel forces it through the porous barrier.
The choice of which filtration technique to use depends on what you are
trying to achieve, but, in general, the following rules apply: if you want the
filtered liquid (filtrate) use gravity filtration. If you want the solid
material use suction filtration.
Thus, if you are trying to remove small amounts of unwanted insoluble
impurities, say, from a solution, gravity filtration using a folded (fluted) filter
paper is often better. In cases where you want to collect a solid material, such
as precipitated or recrystallized product, it is much better to use suction
filtration.
Gravity Filtration
Gravity filtration is a simple technique only requiring a filter funnel, a
piece of filter paper and a vessel, called the filter flask, usually an
Erlenmeyer, for collecting the filtrate. Glass funnels are available in several
sizes. Always use the correct size of filter paper for the filter funnel; after
folding (see below), the filter paper should always be below the rim of the
glass funnel. As a rough guide, use the size of filter paper whose diameter is
about 1 cm less than twice the diameter of the funnel. For example, for a
funnel with a diameter of 6 cm use a filter paper with a diameter of 11 cm.
The purpose of folding or fluting the filter paper is to speed the
filtration by decreasing the area of contact between the paper and the funnel.
Everyone develops their own way of fluting a filter paper, but one way is
shown in Figure 17. Start by folding the paper in half, then in half again, and
then crease each quarter into four more sections with alternate folds in
opposite directions. Pull the paper into a half circle and arrange it into a
pleated fan, ensuring that each fold is in the opposite direction to the previous
one. Pull the sides apart and place the paper into the funnel.
Always support the filter funnel in a metal ring or clamp (Figure 18). It
is very tempting to place the funnel directly into the filter flask with no
additional support; this is bad practice since the whole assembly is top heavy
38

and is easily knocked over. The solution to be filtered is then simply poured
into the filter paper cone, and the filtrate is collected.

Figure 17.: How to flute a filter paper.

Figure 18..: Gravity filtration.

Vacuum Filtration
Carrying out a filtration using vacuum is faster than using gravity
alone, but requires some additional equipment. Since the technique relies on
producing a partial vacuum in the receiving flask, a thick-walled filter flask
39

with a side arm, often called a Buchner flask, must be used, although smaller
quantities can be filtered using a Hirsch tube. The flask should be securely
clamped and attached to the source of vacuum by thick-walled, flexible
pressure tubing. Such tubing is heavy and will almost certainly cause
unsupported flasks to topple over. The source of vacuum in the organic
laboratory is almost invariably the water aspirator and the filter flask should
be protected against suck-back of water by a suitable trap. The most useful
design of trap incorporates a valve for controlling and releasing the vacuum.
Different types of funnel are also used in suction filtration, the usual types are
the so-called Bchner and Hirsch funnels. The Hirsch funnel with its sloping
sides is particularly suited to the collection of smaller amounts of solids. Both
funnels contain a flat perforated plate or filter disk at the bottom, which is
covered with a piece of filter paper. Always use a filter paper of the correct
diameter. Never attempt to use a larger piece and turn up the edges; if the
paper is too large, trim it to size with scissors.

Figure 19.: Suction filtration using (a) a Bchner funnel or (b) a Hirsch
funnel for smaller quantities.
Finally. in order to ensure an adequate seal between the filter funnel and
the flask, the funnel is placed on top of the flask through a Neoprene filter
adapter. The completed assembly for suction filtration using a Bchner or
Hirsch funnel is shown in Figure 19.
Before starting the filtration, wet the filler paper with a little solvent the
same solvent as that used in the solution that is about to be filtered. Turn on
the water aspirator gently, and ensure that the dampened paper is sucked
down flat over the perforated filter disk. Pour the mixture to be filtered onto
the center of the filter paper, and slowly increase the suction. The partial
vacuum in the filter flask results in rapid filtration. With very volatile

40

solvents, do not apply too strong a vacuum, otherwise the filtrate will boil
under the reduced pressure.
When all the liquid has been sucked through, release the vacuum. Wash
the collected solid with a little cold clean solvent and re-apply gentle suction.
Do not wash solids under strong suction because the solvent passes through
too quickly. Another advantage of suction filtration is that continuation of the
suction for a few extra minutes results in fairly effective drying of the solid.
To make this drying as effective as possible, press the solid flat onto the filter
plate using a clean glass stopper, and then maintain the suction to remove the
last traces of solvent, sucking the solid as dry as possible. This process, which
involves drawing large volumes of air through the solid, is a quick way of
drying solids, although it should not be used for compounds which are air or
moisture sensitive.

Figure 20. Some typical sintered glass filter funnels.

After completion of the suction filtration, always release the vacuum
and disconnect the flexible tubing from the filter flask before turning off the
water aspirator.
An alternative to filter paper as a porous barrier is sintered glass. A
variety of filter funnels containing sintered glass disks of varying size and
porosity is commercially available (Figure 20). These make excellent,
although more expensive, alternatives to a Bchner funnel plus filter paper.
In the simplest form, a sintered glass filter funnel has a stem for use with a
Neoprene adapter and filter flask as above. More sophisticated versions
have a standard-tapered ground glass joint with a built-in side arm for
attaching to the aspirator. These can be used for filtering directly into a
standard-tapered joint round-bottomed (not Erlenmeyer unless thick walled)
flask, ready for subsequent evaporation or reaction.
II.7. Extraction of a liquid using a separatory funnel
Discussion

41

Separatory funnels are designed primarily for extracting a compound

from a solution. The liquid to be extracted is often water. Most organic
compounds are more soluble in water-insoluble organic solvents such as
dichloromethane or ether than they are in water. Therefore a compound in
water will readily partition between solvent and water when the two liquids
are mixed together in a separatory funnel (Figure 21).

Figure 21. Distribution of molecules, between water and an organic solvent.

(a) Molecules are in the water layer before being mixed with extracting
solvent. (b) Molecules are distributed between the water and the organic
solvent after the two liquids have been mixed and allowed to separate.
In using ether to extract a compound from water, use three or more
smaller portions rather than one large portion of equal total volume. A total of
300 mL of ether will extract approximately three times as much as a single
100 mL portion.
When one is extracting an organic compound from water, the major
organic solvents used are methylene-chloride, hexane, and diethyl ether. In
contrast, we can extract an organic solvent with water to remove unwanted
components, a process termed washing. If the organic layer contains an acid
to be removed, water or a weak aqueous base solution is used. To remove a
basic compound from an organic layer, a dilute aqueous acid solution is used.
Unfortunately, during the washing of an organic solvent, especially ether
with water, part of the ether or the solvent dissolves in the water and is lost.
To overcome the loss of ether, we should use relatively small volumes of
water. Moreover, the ether layer dissolves water until it contains approximately 78% water by volume. Therefore, a saturated aqueous sodium
chloride solution is often used in place of water for the final washing. The
presence of sodium and chloride ions in the water dramatically reduces water's
ability to dissolve the ether phase, and in fact removes some water already
dissolved in an ether layer.
Apparatus
42

Figure 22. shows the proper setup.

Figure 22. Apparatus for extraction of a liquid by means of a separatory

funnel.
Procedure
The entire series of operational steps described below is normally repeated
two or three times to obtain a thorough extraction of a material from a liquid.
A fresh sample of the extracting solvent has to be used each time the
procedure is repeated.
Step 1: Place the liquid to be extracted in the reparatory funnel (Figure 22).
CAUTION: Be sure the stopcock is closed.
Step 2: Add the extracting solvent to the liquid in the separatory funnel. A
solvent denser than the liquid to be extracted settles to the bottom to form a
lower layer (Figure 23a). If the solvent to be used in extracting the liquid is
less dense than the liquid, it forms a layer on top (Figure 23b). Table 2. lists
various common solvents and their densities.

43

Figure 23.: Extraction of a water solution in a separatory funnel using (a)

a solvent more dense (density greater than 1.0 g/mL) than the solution being
extracted, and (b) a solvent less dense (density less than 1.0 g/mL) than the
solution being extracted
.
Solvent
Approximate density, g/mL
Hexane
0.69
Benzene
0.88
Toluene
0.87
Diethyl-ether
0.71
Water
1.0
Saturated NaCl
1.2
Methylene-chloride
1.34
Chloroform
1.5
Table 2.: Common Solvents Used for Liquid-Liquid Extractions
Step 3: After the separatory funnel is stoppered, slowly turn it upside down and
open the stopcock to let the solvent vapors or gases escape (Figure 24.). It is
wise to place a paper towel over the stem outlet to avoid spattering at this
stage.

44

Figure 24.: Solvent vapors or gases escape from a separatory funnel held
upside down with the stopcock open. CAUTION: Hold glass stopper and
separatory funnel firmly with one hand.
CAUTION: When you are turning the separatory funnel upside down, be sure
to keep a hand over the glass stopper so that it doesn't fall out and release the
liquid inside.
CAUTION: If the vapors are not allowed to escape, gas pressure builds up,
eventually forcing out the stopper and the liquid.
Step 4: Close the stopcock and gently twirl and shake the solution plus the
extraction solvent.
Step 5: Stop twirling or shaking the separatory funnel and repeat Step 3 to
release the vapors.
Step 6: Repeat the sequence of Steps 3, 4, and 5 several times; then go to
Step 7a or 7b as required.
Step 7a: Separation step: If the extraction solvent is more dense than the
liquid being extracted, the extraction solvent is the lower layer. Drain off the
extraction solvent by slowly opening the stopcock after you have removed
the stopper. To remove some traces of water from the organic liquid or to
break up an emulsion if one has formed, drain the lower extracting solvent
layer through a cotton wad placed in a glass funnel, as shown in Figure 25.

45

Figure 25.: Drying the extraction solvent by passing it through a loose wad
of cotton.
Step 7b: Separation step: If the extraction solvent is less dense than the liquid
being extracted, the extraction solvent is the top layer. Drain off the liquid
being extracted by slowly opening the stopcock after you have removed the
stopper. Then drain off the extraction solvent through a cotton plug placed in
a glass funnel, as shown in Figure 25. The cotton plug removes some traces
of water or breaks up an emulsion if one has formed.
II.8. Crystallizations
Introduction
Crystallization is a process used to purify compounds that exist as
solids near room temperature. We shall describe two different methods of
conducting crystallization. The first, crystallization using vacuum filtration, is
the most common method and the only one that can be used if large quantities
are to be purified. However, for purifying small quantities of solids, many
people prefer the second technique, crystallization using centrifugal force, if
the equipment is available.
Both crystallization techniques depend on the generally greater
solubility of solids in hot solvents, compared with cold. A solid is dissolved
in a minimum of boiling solvent and crystallization takes place when the
solution is cooled. Impurities are usually present in only trace amounts and

46

our choice of solvent is made to promote the crystallization of the desired

products while the impurities remain in the cold solvent. The desired crystals
can be separated from the impurities by filtration or centrifugation. Several
repetitions of the process eventually result in pure crystals of the desired
compound.
Choice of Solvents
The choice of solvents to be used in crystallizing a solid is important.
There are only a few solvents used to any degree. The primary rule to
remember is "like dissolves like." In other words, nonpolar solvents dissolve
nonpolar solids and polar solvents dissolve polar compounds. Table 3. is a
list of solvents and their polarities, de-scribed as low, medium, or high.
In some cases a mixed-solvent system is used. In a mixed-solvent
system, a compound is dissolved in a minimum of boiling solvent. Then,
before the solution is cooled, a second miscible solvent of different polarity is
added. Often the original choice of solvents to be used in a crystallization is
made by little more than the-trial-and-error method, using a small amount of
the material to be crystallized in each trial until the proper selection can be
made.
Solvent and type
Polarity
Boiling point (C)
Pure hydrocarbons
Pentane
36
Low
Benzene
80
Low
Toluene
111
Low
Mixture of hydrocarbons
Hexanes
Low
Petroleum ether
30-60
Low
Ligroin
60-90
Low
Alkyl chlorides
Dichloromethane
40
Medium
Chloroform
61
Medium
Carbon tetrachloride
77
Low
Alcohols
Methanol
65
Very high
Ethanol
78
high
Ethers
Diethyl ether
36
Low-medium
Dioxane
101
Low-medium
Ketones
Aceton
56
Medium
47

Esters
Ethyl acetate

66

Medium

Table 3.: Common solvents for use in crystallization

Crystallization using Vacuum filtration
Discussion
Before crystallizing by vacuum filtration, set up a vacuumfiltration apparatus, using a Hirsch funnel or the appropriate size of
Bchner funnel.
Determining the appropriate amount of solvent to dissolve a
given amount of crystals is also a trial-and-error procedure. Remember
that excess solvent can he boiled off and more solvent can always be
added if needed. Furthermore, for convenience you should, if at all
possible, choose a solvent with a low boiling point, since it evaporates
faster when heated on a steam bath. If the boiling point of the solvent is
below 80C, the steam bath is the ideal heating device. If the solvent to be
used has a boiling point above 80C, however, a hot plate is the preferred
heating device.
Procedure
Step I: Set up the vacuum-filtration apparatus.
Step 2: Put the solid in round-bottom flask and take a condenser over it. Add
the minimum amount of solvent that will dissolve the solid at an elevated
temperature. Boil the solvent and solid on a steam bath or a hot plate until the
crystals dissolve in the boiling solvent.
CAUTION: Use a boiling chips.
Step 3: If the solution appears cloudy or contains particles of inert materials,
it should be filtered while it is hot, using gravity filtration
Step 4: Cool the solution, usually by placing the flask in an ice bath.
Step 5: To induce crystallization of an organic solid, you often have to
scratch the beaker or flask on the interior sides and bottom with a glass rod or
metal spatula. If you can't induce crystallization.by scratching, then add a
single crystal of the desired compound, if available, to induce crystallization.
Step 6: After the crystals are formed, remove them from the solution by
vacuum filtration, using a Hirsch or a Bchner funnel.
Special Techniques

48

The crystals can be air-dried by leaving them in the Hirsch funnel (or
Bchner funnel) and sucking air through them. The crystals can also be dried
under IR lamp.
CAUTION: Crystals having a low melting point below 110C cannot be dried
under IR lamp.
II.9. Melting Point
Introduction
By definition, the melting point of a compound is the temperature
at which it changes from solid to liquid form at standard atmospheric
pressure. In practice, the melting point obtained is not a single temperature,
but a range of 0.5 degree or more in which the solid melts. All melting points
are recorded in degrees Celsius (C).

This diagram shows the method of putting a solid into a melting-point tube:
(a) the melting tube; (b) dipping the open end of the tube into a solid; (c)
solid near open end of melting-point tube; (d) dropping the melting-point
tube down a 30- to 50-cm glass tube to strike a hard surface (causing the solid
to fall to the closed end of the melting-point tube); and (e) solid at the bottom
of the melting-point tube.

49

II.10. Thin-layer Chromatography

Introduction
Thin-layer chromatography is useful in analyzing isolated or
commercial products to determine the number and possible identity of
components present. In addition, thin-layer chromatography is often ideal for
following the progress of a reaction, for observing the column
chromatography separation of compounds, or for determining the purity of a
compound after crystallization or distillation.
A chromatography tank is prepared and a solvent (called the running
solvent or eluant), usually chosen by trial and error, is placed in the bottom.
A thin-layer is spotted with a solution containing compounds to be analyzed.
This plate is inserted in the tank to develop a chromatogram.
As the eluant moves up the plate, the various compounds present in
the spot on the plate also migrate upward. The rate at which a compound
migrates up the plate depends on the polarity of the compound, the solid
support, and the eluant. Each compound in the spot is attracted to both the
solid support and the running solvent. If a compound tends to be highly
attracted to the solid support and not the solvent, it spends less time dissolved
in the solvent, and therefore does not migrate up the plate to any degree. On
the other hand, another compound that is more readily dissolved in the
solvent migrates farther up the chromatography plate or paper. Thus the
compounds in a spot are separated from each other on the chromatogram.
Obviously, the degree of separation of components in a mixture depends on
both the solvent and the solid support used. To locate the compounds on the
chromatogram, one must usually develop the chromatogram. When one is
analyzing a dye mixture, the spots are readily visible because of their inherent
color. For detecting compounds that are not colored, however, the
chromatogram can be detected by UV lamp, or can be sprayed with
ninhydrin, placed in iodine vapor, or treated in some other way that will
render the individual spots on the plate visible.
To help identify the compounds present, we can calculate an Rf value
(retardation factor) for each spot present on the chromatogram. In each thinlayer analysis, there is variation in the size of the plate, the thickness of the
solid support, the distance the initial spot is placed from the bottom of the
plate, and the distance the solvent is allowed to migrate up a plate.
Calculating the Rf value for each spot on a chromatogram a value that is
relatively independent of these variables makes possible comparisons
between chromatograms or comparisons with a table of known Rf values. We
can calculate the Rf value by the following formula, in which all distances are
measured from the placement of the initial spot (Figure 26.):

50

Figure 26.: Measurements of solvent and spot distance for determination of

Rf value.
Preparing a Chromatography Tank
Discussion
The size of a chromatography tank is varied according to the size of
the thin-layer plates, whichever are used. Wide-mouthed screw-cap jars or
bottles are ideal for use as chromatography tanks for most experiments. A
filter-paper liner is usually placed in a chromatography tank to help form an
atmosphere saturated with the running solvent, thereby preventing
evaporation of the liquid from the chromatogram.
Low polarity
Cyclohexane
Petroleulm ether
Benzene
Toluene
Carbon tetrachloride

Medium polarity High polarity

Chloroform
Methanol
Dichloromethane Ethanol
Ethyl acetate
Diethyl-ether
1-Buranol
2-Propanol
Table 4.: Solvents used in thin-layer chromatography

Table 4. lists common solvents used for thin-layer and paper

chromatography. The choice of an appropriate solvent system is often a
matter of trial and error. In many cases specialized solvents are specified.
Procedure

51

Select an appropriate size screw-cap jar or bottle for the tank. Line the
inside of the bottle with filter paper (see Figure 27). The liner, on being
saturated with the running solvent, helps maintain an appropriate
environment. The atmosphere should be saturated to prevent evaporation of
solvent from the surface of the chromatograms as they are developed.
After selecting a running solvent, place it in the bottom of the bottle,
replace the top, and shake the whole to saturate the filter-paper liner. The
depth of the liquid in the bottle should not exceed 0.5 cm.
CAUTION: The surface of the running solvent must not be above the
initial spot on the paper or thin-layer chromatogram.

Figure 27.: A chromatography tank

Running a Chromatogram; Determining Rf Values
Discussion
After you prepare the chromatography tank, select the proper solid
support for thin-layer plate and spot the chromatogram, run the
chromatogram. After you develop the chromatogram, determine the Rf
values.
Procedure
The procedure for running a chromatogram is as follows.
Step 1: Place the paper or thin-layer chromatogram in the chromatographic
tank previously prepared and replace the lid.
CAUTION: Be sure that the bottom of the chromatogram does not touch the
filter-paper liner in the tank.
CAUTION: Be sure that the initial spot is above the level of the solvent in the
tank when the end of the chromatogram is resting on the bottom of the tank.

52

Step 2: Remove the chromatogram before the solvent reaches the top of the
paper, silica gel G, or other solid support, and immediately mark the highest
point the solvent has reached.
Step 3: If necessary, develop the chromatogram as prescribed in the
experiment and mark the location of the center of each spot .
Step 4: Measure the distance (dsolvent) between the initial spot and the highest
point reached by the solvent and record the measurement (ds). Measure the
distances between the initial spot and the center of each spot, dA, dB, dC, ... ,
representing the location of each compound separated. Record each distance
(Figure 28.).
Step 5: Calculate the Rf (retardation factor) for each spot and record.
Determine the Rf value for each spot by dividing the distance the spot has
moved from the initial site by the solvent front distance, dS. For example, the
calculated value for the Rf for compound A in Figure T17.7 is:

Figure 28.: A developed chromatogram, showing initial and other spots and
all measurements that must be made and recorded.
Developing the Chromatogram
Discussion
If colorless compounds are chromatographed, they can be detected
under UV lamp, or they must be treated after the chromatogram has been run,
to develop a color. Only when each spot representing a compound has been
treated to reveal its location can you determine the Rf values and analyze the
chromatogram.
II. 11. Column Chromatography

53

Introduction
Column chromatography is a technique for separating large amounts
of compounds. It relies on the relative polarities of the molecules involved.
The investigator fills a glass tube with a fine-powdered polar material (solid
support) such as alumina or silica gel in an organic solvent (Figure 29).
A solution containing the compound is applied to the surface of the
solid support and then an organic solvent is drained through the column. The
liquid drained through the column is collected as a series of fractions in
separate containers. The passing of the solvent through the column is
continued until all the desired compounds present in the solution placed on
the packed column have been eluted from the column and collected as
separate fractions.
The rate at which a compound is eluted from the column depends on
its polarity, the polarity of the solid support, and the polarity of the solvent
used to elute the compounds from a column. A compound is attracted both to
the solid support, such as alumina, and to a solvent such as methylene
chloride. If it is more attracted to the solid support than to the solvent, it
spends less time in the solvent and therefore migrates down the column
slowly. In contrast, a compound that is more attracted to the solvent spends
more time in the eluting solvent and therefore migrates down the column
more rapidly. It exits from the column long before the compound more
tightly bound to the solid support material. Often the initial eluting solvent,
which is relatively nonpolar, is exchanged for a more polar solvent during
subsequent stages of elution of the column. The compounds actually form
separate bands on the column. Some of these bands may be visible, or may be
made visible with ultraviolet light. These bands move down and out of the
column as solvent passes through the solid support.

54

Figure 29.: Column packed with solid support in an organic solvent.

Packing a Chromatography Column
Discussion
There are two basic types of chromatography columns (Figure 30).
The preferred one (Figure 30a) contains a stopcock for controlling flow of the
solvent. A buret can be used in many instances. The second model has a
solvent-insoluble plastic tube (usually Tygon) attached to the exit of the
column that can be closed and opened by means of a screw clamp.

Figure 30.: Chromatography columns: (a) with stopcock; (b) with Tygon
tube and screw clamp.

55

The two methods of packing a column are dry pack and slurry pack.
Dry pack is the easier method.
Procedure
Dry Pack Place a small wad of glass wool in the bottom of the
chromatography column. Fill the column three-fourths full of eluting solvent
(Figure 31a).
CAUTION: The wad of glass wool must be thick enough to prevent
passage of solid support material, but not thick enough to cause air to be
trapped in large amounts.
Drain a small amount of solvent from the column. Remove air bubbles from
the glass wool plug by tamping it with a solid glass rod.
CAUTION: The glass wool must be free of air bubbles before you
continue.
Place a powder funnel on top of the column and add a thin layer of sand on
top of the glass wool. Then slowly add solid support, such as alumina or
silica gel, as shown in Figure 31b. To help pack the column, occasionally
drain small amounts of the solvent from the column. Also gently tap the
column with a piece of pressure tubing to help the solid support settle. The
column support must be packed gently until it no longer settles. The final
column of solid support should be 10 to 15 times as tall as it is wide.
CAUTION: Never drain out solvent to the extent that the surface of
the solid support is exposed.
A small layer of sand is normally placed on top of the solid support to
protect the surface. It too must be below the surface of the eluting solvent.

56

Figure 31.: Chromatography columns: (a) solvent added to column; (b) solid
support being added to column.
CAUTION: The top of the solid support must be level and no bubbles
should be in the column. Bubbles or a top that is slanted cause the
compounds, in the form of separate bands moving down the column, to be
deformed and thus become mixed as they are eluted by the solvent (Figure
32).

57

Figure 32.: Two things to avoid. (a) Deformed bonds of compounds on a

column caused by a slanted top. (b) A large air bubble in the solid support.
Slurry Pack This method is identical to the dry pack, except that the
solid support is placed in the solvent to make a thin slurry. This slurry is then
added to the column filled with solvent. In all other respects, slurry-pack
procedures are identical to dry-pack procedures.
Placing a Compound Mixture to Be Separated on a Column; Eluting
Compounds

Procedure
Step 1: Drain the solvent from the column until the solvent layer is level with
the surface of the solid support.
Step 2: Dissolve compounds either in a minimum of the same solvent used in
packing the column, or in a second prescribed solvent. [Note: If compounds
are al-ready liquid, place the liquid directly on the column (Step 3).]
Step 3: Carefully pipet or pour solution from Step 2 onto surface of column.
CAUTION: When placing solution containing compounds in column, do not
disturb surface.
Step 4: Drain solvent from column until liquid surface in column is again
even with the top of the solid support.
Step 5: With as little turbulence as possible, add eluting solvent to column
until it is nearly filled. The initial eluting solvent is usually the same as that in
which solid support was originally packed. [Note: Maintain a uniform solvent
58

head (Figure 33) to create enough force to provide an even flow rate. A
solvent head is the solvent above the solid support in the chromatography
column.] To maintain a solvent head for a large chromatography column,
place a capped separatory funnel containing solvent at the top of the column,
as shown in Figure 33. Then open the stopcock. [Note: As the solvent drains
from the column in Step 6, the level of the solvent drops lower than the
bottom of the separatory funnel stem. When this happens, a bubble of air
enters the separatory funnel through the stem and some solvent drains out.
Bubbles of air continue to enter the funnel and solvent drains out until the
bottom of the separatory funnel stem is again below the surface of the solvent
in the column.]
Step 6: Drain the solvent from the column at a rate of 15 drops per second.
Change the collection vessel as directed, or every time a volume of solvent
equal to 5% of the volume of the support has exited from the column.
CAUTION: Always add more eluant solvent before the level of the
solvent reaches , the upper surface of the sand layer.
Step 7: [Note: This step is used only if there is to be a change of eluting
solvents.] Drain out all solvent until liquid surface is even with the upper
surface of the sand in the top of the column. Repeat Step 5 to add new eluting
solvent. Then proceed to Step 6.
Step 8: Evaporate to dryness each solvent fraction collected .

59

Figure 33.: Chromatography column with separatory funnel to maintain a

solvent head

Monitoring Column Chromatography

Discussion
It is necessary to monitor a chromatography column to determine
whether all compounds have been eluted, whether separation has occurred,
60

and to determine which evaporated fractions may be combined. The column

chromatography procedure is usually, but not always, continued until all
materials are eluted. An individual compound rarely comes off in a single
fraction. Therefore fractions are often combined until all of a pure component
is collected together. Unfortunately, bands often overlap on the column and
therefore some fractions can contain two components. These fractions must
not be combined with the fractions containing only one compound.
Let us consider a standard column chromatography of two
compounds, A and B.
Fractions 1-6: No compounds
Fractions 7-15: Compound A
Fractions 16-20: Mixture of compounds A and B
Fractions 21-28: Compound B
Fractions 29-30:No compounds
Therefore fractions 7-15 are combined to obtain compound A.
Fractions 21-28 are combined to give compound B, and fractions 16-20 are
either chromatographed again or discarded.
Procedure
Normally we use thin-layer chromatography to monitor a column. We
prepare microscope slides having a solid support and solvent identical to that
used in the column chromatography. We spot a sample of starting material,
the fraction to he analyzed, and a standard (If available) on the thin-layer
plate for analysis
CAUTION: Since a solvent fraction is often very dilute, it usually requires
evaporation to near dryness before it can be analyzed.

61

III. Preparation and reaction of organic compounds

The goal of this part of organic laboratory practice is to use some
principle techniques of organic chemistry (crystallization, distillation,
filtration, thin-layer chromatography, determination of melting point).
1. Crystallization
Objectives:
To demonstrate the crystallization and learn the techniques of
crystallization
Chemicals and equipment
Bunsen burner
Round-bottomed flask
Filter paper
Acetanilide (2g)
Erlenmeyer flask
Capillary

Screen
Condenser
Funnel
Benzanilide (2g)
Bchner funnel
Water

Water bath
Boiling chips
Clamp
Charcoal
Filter flask
Methanol

1.1 Crystallization from water

Procedure
Place the acetanilide in a round-bottomed flask (50 cm3), and add 10
cm3 water and some boiling chips into the flask. Take a condenser into the
flask, and bring the mixture to the boiling point by heating with Bunsen
burner. Add solvent dropwise through the condenser with continued heating
until the material has solved. Add small amount of charcoal to remove
colored minor impurities. Filter hot solution into an Erlenmeyer flask (use
gravity filtration). Cool the clear solution to allow crystallization. Collect the
crystals by filtration and dry it on air. Determine the weight and the melting
point of crystals, and calculate the yield of crystallization.
1.2 Crystallization from methanol
Procedure
Place the benzanilide in a round-bottomed flask (50 cm3), and add 10
cm methanol and some boiling chips into the flask. Take a condenser into
the flask, and bring the mixture to the boiling point.
3

62

CAUTION: Methanol is flammable organic solvent. Do not use

Bunsen burner in the same hood. Use hot water bath heating by hot plate
as heat source.
Add solvent dropwise through the condenser with continued heating
until the material has solved. Add small amount of charcoal to remove
colored minor impurities. Filter a hot solution into an Erlenmeyer flask (use
gravity filtration). Cool the clear solution to allow crystallization. Collect the
crystals by filtration and dry it on air. Determine the weight and the melting
point of crystals, and calculate the yield of crystallization.
2. Controlling of purity by thin-layer chromatography, (TLC) and
determination of melting point. Distillation.
Objectives:
1. To demonstrate the simple and vacuum distillation.
2. To learn the techniques of distillation.
3. To demonstrate the TLC and learn the techniques of TLC.

Chemicals and equipment

Water
Boiling chips
Distillation head
Acetanilide
Ethyl-acetate

Water bath
Clamps
Condenser
Benzanilide
Running tank

Vacuum capillary
Round-bottomed flask
Vacuum adapter
Hexane

2.1 Vacuum and simple distillation

Procedure
Assemble a vacuum distillation apparatus and place 100 cm3 water
into the distillation flask.
CAUTION: Check intact of distillation apparatus. Grease lightly all
ground-glass joints with stopcock grease before use. Use safety glasses over
the distillation.
Turn on water to condenser. Turn water aspirator on by opening valve
to maximum water flow, and attach system to water aspirator. Raise heating
mantle and slowly raise temperature until distillation begins.
CAUTION: Raising the temperature too rapidly can cause
overheating and decomposition of material, creating a fog in the distillation
flask. Never evaporate all of the solution.
63

Record the exact boiling point and pressure (C/Hgmm).

Shut-down procedure: remove and turn off heat source, when 30 cm3 of
water has distillated. Turn off vacuum by removing of vacuum tube from the
vacuum adapter, and wait for the pressure of system has reached equilibrium
with atmospheric pressure.
Resume distillation at atmospheric pressure until small amount of
water (30 cm3) remains in distillation flask. Turn off heat source, and
disassemble your apparatus after cool down.
2.2 Thin-layer chromatography (TLC)
Control the purity of acetamide and benzamide by TLC, and
determine the Rf values.
Procedure
Solve some crystals in acetone and spot the chromatogram. Run the
chromatogram, using hexane : ethyl-acetate = 3:1 as running solvent. After
you develop the chromatogram, detect spots by UV lamp, and determine the
Rf values.
3. Isolation of nicotine from tobacco leaves.
Nicotine is a major alkaloid of tobacco (Nicotiana tabacum). In pure
form is colorless poisonous oil. It is used as insecticide. Nicotine is soluble in
water, mild base and evaporates with steam. Nicotine-dipicrate can use for
precipitate from water, because of its low solubility in water.

H
N
CH3

N
H

nicotine
C10H14N2
Mol. Wt.: 162.23
Bp: 246-247C

NH x
CH3

O2N

NO2

NO2

nicotine-dipicrate
C22H20N8O14
Mol. Wt.: 620.45
Mp: 218C

Background to the Procedure

Tabacco leaves contain nicotine as a salt of malic acid, which is
soluble in water. Nicotine can release from this salt with weak base (MgO),
and then separate with steam distillation, and isolate its dipicrate.

64

Chemicals and equipment

Water
Magnesium-oxide

Tabacco leaves
Picric acid

Sodium-chloride
Steam distillation apparatus

Procedure:
To the aqueous solution of sodium-chloride (25g NaCl in 75 cm3
water) in steam distillation flask add 5g of tobacco leaves, and shake it.
While it is standing, assemble the steam distillation apparatus, and start to
heat the boiler. After 30 minutes add the suspension of magnesium-oxide
(1g) in water (25 cm3) to the tobacco leaves, and attach the distillation flask
to the steam distillation apparatus. Start the distillation and collect ca. 100
cm3 of distillate. This distillate contains nicotine base.
Add the saturated aqueous solution of picric acid (0,6g picric acid).
Nicotine-dipicrate crystallizes as yellow crystals. Put it to an ice bath, and
then filter, washed with water (2x20 cm3), and dry under IR lamp. Measure
the mass of product, determine nicotine contents of tobacco.
Measure the melting point of nicotine-dipicrate and determine the Rf
value, using next procedure: Solve a small amount of dipicrate (0,01g) in hot
water in test tube, and add aqueous solution of ammonia to it until the pH
turn to alkaline. Put 1cm3 of ether to the test tube and shake it. Spot the
chromatogram, and develop it using toluene : methanol = 4:1 as running
solvent. Use Dragendorff-reagent to detect of alkaloids.
4. Isolation of caffeine from tea leaves
Caffeine is a major alkaloid of tea (Camellia sinensis). The drug
caffeine is used as a cardiac, respiratory, and mental stimulant. It is also often
used as a vascular cephalagic, especially for migraine headaches. Caffeine
belongs to a group of alkaloid compounds called the xanthines.
O
H 3C

CH3
N

N N
CH3
caffeine
Besides being found in tea leaves, caffeine is present in coffee, kola
nuts and cocoa beans. As much as 5% by weight of the leaf material in tea
plants consist of caffeine.
O

65

Background to the Procedure

Tea leaves consist mostly of cellulose a water-insoluble polymer of
glucose, tannins (phenolic compounds, compounds that have an -OH directly
bonded to an aromatic ring) and a small amount of chlorophyll.
The idea in this experiment is to extract the water soluble materials in
the tea leaves into hot water. The hot solution is allowed to cool and the
caffeine is then extracted from the water with dichloromethane (methylene
chloride). Since caffeine is more soluble in dichloromethane than it is in
water, it readily dissolves in the dichloromethane. However, the tannins are
slightly soluble in the dichloromethane. But we want to separate the caffeine
from the tannins by having the caffeine dissolve in the dichloromethane and
the tannins remain in the water. We can do this by taking advantage of the
fact that phenols are acidic enough to be converted to their salts
(deprotonation of the -OH group) by reacting with sodium carbonate. So, we
will add sodium carbonate to the water and the tannins will be converted to
phenolic anions, which are not soluble in the dichloromethane but are soluble
in highly polar water.
Consequently, as you extract the caffeine from the water into the
dichloromethane do not shake the separatory funnel vigorously.
Chemicals and equipment
Water
Sodium-carbonate
Separatory funnel

Ice bath
Dichloromethane
Erlenmeyer flask

Tea leaves
Magnesium-sulphate
Baker

Procedure:
Open two tea bags and place tea leaves (3g) into 100 cm3 hot water,
and leave it to stand for ten minutes. Subsequently separate the solution from
teal leaves and cool it to room temperature with ice bath. Add 2,0g of
sodium-carbonate to the solution, and extract with dichloromethane (2x20
cm3). Dry the organic solvent under anhydrous magnesium-sulphate, and
evaporate the solvent. Determine the caffeine content of the tea leaves.
Control the purity of caffeine by TLC (running solvent: toluene: methanol =
4:1) and determine the melting point of caffeine. Use ammoniummolibdenate for detect the TLC of caffeine.

66

5. Separation of organic compounds with liquid-liquid extraction.

Identification of hydrocarbons
5.1. Separation of organic compounds with liquid-liquid extraction.
The use of a separatory funnel is a common operation in organic
chemistry. In the course of separating a mixture (or working up a raction)
chemists frequently take advantage of the acidic or basic properties of a solute
to partition it between an organic phase and an aqueous phase.
1,3-dinitrobenzene (1,3-DNB) could be transformed into 3-nitroaniline (3-NA) by partial reduction. We can separate the product of this
reaction from starting material, because there are different acidic-basic
properties:
NO2

NH2

NO2

(NH4)2Sx
reduction

natural
organic soluble

NH3
acid
NO2

NO2

basic
organic soluble

acidic
water soluble

Chemicals and equipment

Water
10 % HCl
Separatory funnel

Dichloromethane
10% NaOH
Erlenmeyer flask

Mixture of 1,3-DNB and 3-NA

Methanol
Round-bottom flask

Procedure:
To 20 cm3 of dichloromethane in Erlenmeyer flask add a mixture of
1,3-dinitrobenzene and 3-nitro-aniline (0,4g) and spot thin-layer
chromatogram (do not develop the chromatogram). Pure the solution into
separatory funnel, and extract with 10% HCl solution (10 cm3). Separate the
organic (lower phase) and water phase (upper phase). Pure the organic phase
again into separatory funnel, extract with water (10 cm3), and separate them.
Isolation of 3-nitro-aniline (3-NA)
Take the Erlenmeyer flask with combined inorganic phases (water) in
an ice bath, and add 10% NaOH solution into the flask until the solid product

67

is precipitated. Filter the solid product (use dipolder), wash with water (2
cm3) and dry on air. Determine the yield of separation and melting point.
Isolation of 1,3-dinitrobenzene (1,3-DNB)
Dry the organic phase under MgSO4. Filter the solvent into roundbottom flask and evaporate the solvent using rotatory evaporator. Solve the
residue in methanol (3 cm3) and add 3 cm3 of water to it. Filter the
precipitation (use dipolder), wash with water (2cm3), and dry on air.
Determine the yield of separation and melting point.
Check the purity of compounds by TLC. Spot the chromatogram (to
the same plate as earlier; see the scheme), and develop the chromatogram
using ethyl-acetate : toluene = 4:1 as running solvent. Determine the Rf value
of compounds.

3-NA

mixture

1,3-DNB

5.2. Identification of hydrocarbons

Hydrocarbons with multiple bonds (unsaturated hydrocarbons except
most cycloalkanes) react with bromine. Alkenes and alkynes undergo an
addition reaction with bromine.
Br
R
Br2
R
R
R
Br
The double bond of an alkene becomes a single bond and one bromine
atom becomes bonded to each of the carbons that shared the double bond. No
other product is formed; the alkene and bromine simply add together, which
is why its called an addition reaction. The triple bond of an alkyne also
undergoes an addition becoming a single bond, but in this case each of the
carbons that were joined by the triple bond will now hold two bromine atoms.

68

Procedure:
In a hood, 0.1g or 2 drops of the compound is added to 0,5 cm3 of
dichloromethane, and a 2% solution of bromine in dichloromethane is added
drop by drop, with shaking, until the bromine color persists.
Bromine has a reddish-brown color. All of the other substances in
these reactions are colorless. So, when bromine is added to an alkene or
alkyne the red-brown color dissipates quickly, often almost instantly.
Bromine can also react with an alkane, but this reaction requires heat
or ultraviolet light to be successful, and the reaction is a substitution, not an
addition: a hydrogen is replaced by a bromine and hydrogen bromide is a
byproduct.
R

Br2

+ HBr

Br

Since this reaction does not take place in the absence of ultraviolet
light or heat, if bromine is added to an alkane under these conditions (room
temperature and no sunlight or other source of UV) the reddish-brown color
of bromine will persist. Aromatic compounds with alkyl side chain also react
under this reaction condition.
Procedure:
In a hood, 0.2g or 5 drops of the compound is added to 1 cm3 of
dichloromethane, and 6 drops of a 2% solution of bromine in
dichloromethane is added. Light the test tube with UV lamp for two minutes.
Check the evolution of hydrogen bromide gas with wet pH indicator paper.
with UV light
color of pH indicator

without UV light
Hexane
Cyclohexene
Benzene
Toluene
Styrene
Unknown

Aromatic compounds can be alkylated with alkyl-halogenides in the

presence of Lewis acids (AlCl3) (Friedel-Crafts reaction). Using chloroform
as a reagent furnish triaryl-methane, which forms a color salt.

69

R
3R

+ CHCl3

AlCl3

AlCl3
R

HAlCl3
R

Procedure:
To 0,5 cm3 of dry chloroform in a test tube add 2 drops or 0.01g of the
compound. Mix thoroughly, and incline the test tube so as to moisten the
wall. Then add some crystals of anhydrous aluminum chloride so that some
of the powder strikes the side of the test tube. Note the color of the powder on
the side, as well as the solution. Discussion: The colors produced by the
reaction of aromatic compounds with chloroform and aluminum chloride are
quite characteristic.
Compounds
Hexane
Benzene
Toluene
Styrene
Unknown

Color

Dilute aqueous potassium permanganate oxidizes alkenes to geminal

diols. [Diol means two -OH groups. Geminal means on adjacent carbons].
Alkynes are oxidized to geminal diketones. (Bayer probe)
OH
R KMnO4
R
R
R
OH

O
R

KMnO4

O
In these processes the purple potassium permanganate is reduced to a
brown precipitate of manganese dioxide. Since the potassium permanganate
is soluble in water, but neither the water nor the potassium permanganate are
soluble in the hydrocarbon, reaction takes place at the water-hydrocarbon
interface, and is somewhat slow. Consequently, it may take several minutes
for a brown precipitate to form. Alkanes and aromatic rings are unreactive
toward dilute aqueous potassium permanganate. Easily oxidized compounds
(aldehydes; formic acid and its esters; alcohols with trace impurities; phenols
and aryl amines; mercaptans and thioethers) also give a positive test, but

70

carbonyl compounds which decolorize bromine/carbon tetrachloride usually

give a negative test.
Procedure:
To 0,5 cm3 of acetone (distillated from KMnO4) add 0.01g or 2 drops of
the compounds. Then add 1 drop of a 1% aqueous potassium permanganate
solution and shaking until the purple color of the permanganate persists. The
disappearance of the purple color and the appearance of a brown suspension
is a positive test.
Compounds
Hexane
Cyclohexane
Benzene
Toluene
Styrene
Unknown

6.

Preparation and purification of terc-buthyl-chloride.

Identification of organic halides (test tube reactions)

6.1. Preparation and purification of terc-buthyl-chloride

Synthesis of alkyl-halogenids by nucleophilic substitution (SN1)

H3C

CH3
OH
CH3

cc. HCl
water

C4H10O
Mol. Wt.: 74.12
Mp: 25C
Bp: 83C
d: 0.789 g/cm3

H3C

CH3
Cl
CH3

C4H9Cl
Mol. Wt.: 92.57
Mp: - 27C
Bp: 51C
d: 0.847 g/cm3

Chemicals and equipment

terc-butanol
Sodium-kloride

cc. HCl
Calcium-kloride

71

Sodium-hydrogencarbonate
Separatory funnel

Simple distillation
apparatus

Erlenmeyer flask

Procedure:
Shake the mixture of terc-butanol (10cm3) and cc. HCl (25cm3) in
separatory funnel for five minutes, and then leave to separate the phases.
Separate the water phase (lower phase), and wash the organic phase with
saturated aqueous solution of NaHCO3, then saturated aqueous solution of
NaCl, and dry over CaCl2.
Caution: If separation isnt successful, you can experience evolution
of gas (CO2), which can cause overpressure in seperatory funnel.
While the terc-buthyl-chloride is dried, assemble a simple distillation
apparatus, and after filtration purify the terc-buthyl-chloride by fractional
distillation.
6.2. Identification of organic halides
If a copper wire is heated in the flame of Bunsen burner a green color
is usually imparted to the flame in the presence of halogenic compounds
(Beilstein test).
Procedure:
Heat the tip of a copper wire in a burner flame until there is no further
coloration of the flame. Let the wire cool slightly, then dip it into the
unknown (solid or liquid) and again, heat it in the flame. A green flash is
indicative of chlorine, bromine, and iodine; fluorine is not detected because
copper fluoride is not volatile. The Beilstein test is very sensitive, thus
halogen-containing impurities may give misleading results.
Compuond
n-Buthyl-bromide
Allyl-bromide
Chlorbenzene

Color of flame

Alkyl-, allyl- and benzyl-halogenides react in

substitution reaction with alcoholic solution of silver nitrate
R OC2H5
R Hlg

AgNO3

HNO3

nucleophilic

+ AgHlg

C2H5OH
R ONO2 +

Procedure:
72

C2H5OH

+ AgHlg

Place 2 drops of compound into a test tube. Add 0.5 cm3 of a saturated
ethanolic silver nitrate solution to the material in each test tube. After the
addition, shake the test tube vigorously to ensure adequate mixing of the
compound and the solution. Record the time required for some precipitate to
form. If no precipitate is seen after 5 minutes, heat the solution on the steam
bath for approximately 5 minutes. Note whether some precipitate forms in the
test tube.
If there is some precipitate, note its color. Add 2 drops of 5% nitric
acid, and note if the precipitate dissolves. Silver halides are insoluble in
dilute nitric acid; silver salts of organic acids are soluble. Acyl-halide,
carboxylic acid and -halo-ethers also give positive probe
Compound
n-Buthyl-bromide
sec-Buthyl-bromide
terc-Buthyl-bromide
Allyl-bromide
Benzyl-chloride
Chlorbenzene

7. Separation of acetanilide and m-dinitrobenzene by column

chromatography

Chemicals and equipment

mixture of acetanilide and
m-dinitrobenzene

Hexane Ethyl
acetate = 2:1

Silicagel

Procedure:
Solve 0,1 g of mixture of acetanilide and m-dinitrobenzene in small
amount (0,5 cm3) of eluent (if is it necessary you can add some drops of
EtOAc) Determine the components of mixture by thin-layer chromatography
using hexane ethyl-acetate = 2:1 as running solvent (detection: under UV
light).
Make column from 5g silica gel using Slurry pack method (the
silica gel is placed into the eluent (hexane ethyl-acetate = 2:1) to make a
thin slurry. This slurry is then added to the column filled with solvent).
Place the solution of the mixture carefully to the column using
Pasteur-pipette. Elute the components from the column with eluent (hexane
73

ethyl-acetate = 2:1) (collect fractions with 4 cm3 volume). Determine the

components of all the fractions by TLC.

Isolation of S-(+)-Carvone from caraway.

Carvone is a member of a family of chemicals called terpenoids.
Carvone is found naturally in many essential oils, but is most abundant in the
oils from seeds of caraway (Carum Carvi). Carvone forms two mirror image
forms or enantiomers: S-(+)-carvone smells like caraway. Its mirror image,
R-()-carvone, smells like spearmint. The fact that the two enantiomers are
perceived as smelling differently is proof that olfactory receptors must
contain chiral groups, allowing them to respond more strongly to one
enantiomer than to the other. Not all enantiomers have distinguishable
odours.
O

H
S-(+)-carvone

R -(-)-carvone

S-(+)-Carvone is the principal constituent (50-70%) of the oil from

caraway seeds (Carum carvi), which is produced on a scale of about 10
tonnes per year. It also occurs to the extent of about 40-60% in dill seed oil
(from Anethum graveolens), and also in mandarin orange peel oil. R-()Carvone is present at levels greater than 51% in spearmint oil (Mentha
spicata), which is produced on a scale of around 1500 tonnes annually. This
isomer also occurs in kuromoji oil. Some oils, like gingergrass oil, contain a
mixture of both enantiomers. Many other natural oils, for example
peppermint oil, contain lower concentrations of carvones.
Objectives: Isolation of carvone from caraway by steam distillation
and transformation into 2,4-dinitrophenylhydrazone derivative.
NO2
O

N
2,4-DNP

N
H

NO2

H
C10H14O
Mol. Wt.: 150.22
b.p.: 227-230 C

C16H18N4O4
Mol. Wt.: 330.34
o.p.: 189 C

74

Isolation of Carvone:
To the caraway (25 g) in steam distillation flask add 100 cm3 water
and attach the distillation flask to the steam distillation apparatus. Start the
distillation and collect ca. 150 cm3 of distillate. This distillate contains
carvone. Extract this distillate with dichloromethane (3x20 cm3), dry the
organic solvent under anhydrous magnesium-sulphate, and evaporate the
solvent.
Preparation of 2,4-dinitrophenylhydrazine reagent
On a steam bath solve 0,2 g of 2,4-dinitrophenylhydrazine in 85%
phosphoric acid and dilute with ethanol (1,5 cm3).
Preparation of 2,4-dinitrophenylhydrazone derivative of carvone
Add 2,4-dinitrophenylhydrazine reagent to the solution of carvone
(0,3 g) in ethanol (2,5 cm3). The 2,4-dinitrophenylhydrazone derivative of
carvone crystallizes as red crystals. Filter off the crystals and washed with
water (2x20 cm3). Control the purity of crystals by measurement of melting
point.

Isolation of Piperine from black pepper.

Piperine is the alkaloid responsible for the taste and smell of black
pepper. It has also been used in some forms of traditional medicine and as an
insecticide.
O
O

C17H19NO3
Mol. Wt.: 285.34
m.p.: 130-132 C

Piperine has been found to inhibit human P-glycoprotein and

CYP3A4, enzymes important for the metabolism and transport of xenobiotics
and metabolites. In animal studies, piperine also inhibited other enzymes
important in drug metabolism.
Experiment: Place the black pepper (10g) in a round-bottomed flask
(50 cm3), and add 10 cm3 dichloromethane and some boiling chips into the
flask. Take a condenser into the flask, and reflux by heating with water bath
for 20 minutes. Cool it down to room temperature, filter off the pepper and
evaporate the solvent. Add ether (5 cm3) to the residue and evaporate it. The
residue is crystallized from ether (5 cm3). Filter off the yellow crystals and
dry it on air. Control the purity of piperine by measure of melting point and
calculate the piperine content of black pepper.

75

Hydrolysis of Piperine in alkaline media.

The hydrolysis of piperine with potassium hydroxide furnishes
piperidine and piperinic acid:
O
O

O
N

KOH

OH

C12H10O4
Mol. Wt.: 218.21
m.p.: 216-217 C

C17H19NO3
Mol. Wt.: 285.34
m.p.: 130-132 C

+
N
H2Cl

C5H12ClN
Mol. Wt.: 121.61
m.p.: 216-221 C

Experiment: Place the piperine (1 g) in a round-bottomed flask (50 cm3),

and add 10 cm3 of 10 % alcoholic solution of KOH and some boiling chips
into the flask. Take a condenser into the flask, and reflux by heating with
water bath for 1 hours. Then evaporate ethanol using normal distillation
apparatus, while the round bottom flask is placed into ice bath. While this
alcoholic solution contains piperine, potassium salt of pipperinic acid is
crystallized in distillation flask. Suspend this salt in a hot water and add cc.
HCl solution to it. Filter off the precipitated piperinic acid and dry it under IR
lamp.
Add cc. HCl acid to the alcoholic solution of piperidine and evaporate the
solvent using rotatory evaporator. Measure the melting point of both
compounds.

76

Identification of hydroxyl derivatives of hydrocarbons.

1. Lucas test of alcohols
Aliphatic alcohols react with different rate with the Lucas reagent
(concentrated hydrochloric acid solution of zinc chloride) depending on the
order of substitution of the carbon carrying the OH group. The reaction has
SN1 mechanism for tertiary and secondary alcohols. Thus tertiary alcohols
react immediately, while secondary alcohols react much slower (1-5 min) and
heating can be applied to make the reaction faster. While primary aliphatic
alchols do not react within one hour without heating. During a positive test,
the solution becomes cloudy due to the formation of the alkyl chloride which
is not soluable in aqueous HCl and a separate layer may appear if larger
amounts of reagents are applied.
R O
H

ZnCl2

R +

R O ZnCl2
H

[Zn(OH)Cl2]

ZnCl2

H2O

HCl
RCl

Experiment: To 10 drops of the Lucas reagent, two drops of the studied

alcohol are added, shaked and allowed to stand at room temperature. The
changes were monitored. If no changes were observed, heat it up to 50 C
and keep at that temperature for 15 minutes.
Butane1-ol

Butane2-ol

tercButanol

Benzylalcohol

unknown

Structure of
alcohol
Observations
Remarks: The alcohol should dissolve in hydrochloric acid and hence this
test is reliable up to six carbon-atom alcohols. Due to mechanism of the
reaction, benzyl and allyl alcohols reacts the same manner like tertiary
alcohols.

77

2. Oxidation of alcohols by Jones reagent

Various oxidizing agents based on Cr (VI) have been used to oxidize
secondary alcohols to ketones. The most commonly used reagent is chromic
acid (H2CrO4). Chromic acid is usually prepared by adding Cr(VI) oxide
(CrO3) or sodium dichromate (Na2Cr2O7) to aqueous sulfuric acid. As
chromic acid oxidizes the alcohol to the ketone, chromium is reduced from
the +6 oxidation state (H2CrO4) to the +3 oxidation state (Cr3+). The use of
CrO3 in aqueous acetone is usually called the Jones oxidation (or oxidation
by the Jones reagent).
OH
3
R

O
+ 2 H2CrO4 + 6 H

+ 2 Cr

3
R

+ 8 H2O

Oxidation of Primary Alcohols to Aldehydes

The oxidation of aldehydes to carboxylic acids in aqueous solutions is
easier than oxidation of primary alcohols to aldehydes; thus it is difficult to
stop the oxidation at the aldehyde stage.
R

OH

ox.

R CHO

ox.
R

OH

Primary and secondary alcohols are rapidly oxidized by a solution of CrO3 in

aqueous sulfuric acid. Chromic oxide (CrO3) dissolves in aqueous sulfuric
acid to give a clear orange solution containing Cr2072- ions. A positive test is
indicated when this clear orange solution becomes opaque and takes on a
greenish cast within 2 s.
Experiment: To one drop of the studied alcohol, eigth drops of acetone
(distilled over KMnO4) are added, and then carefully one drop of Jones
reagent.
Buta
ne-1ol

2methylprop
ane-1-ol

Glyce
rol

Structure
of
alcohol
Observati
ons

78

Phen
ol

tercButa
nol

Benzylalc
ohol

unkno
wn

3. Reaction of diols or poliols with Cu (II) ions

Vicinal diols, acting as bidendate ligands, form a complex with copper
ions, whose colour can be distinguisehd well from the colour of the complex
formed from Cu(II) hydroxyde due to the excess NaOH.
OH
OH

Cu2

Propan
e-1-ol

O
Cu
O
H

HO
HO

Propan
e-1,2diol

1Methoxypropa
ne-2-ol

Glycer
ol

H
O
O

Dgluco
se

Unkno
wn

Structure
of alcohol
Observatio
ns

Experiment: 2 ml water and one drop of 10 % CuSO4 solution are added

to approximately 30 mg or five drops of alcohols followed by the addition of
8-10 drops of 8% NaOH solution.
4. Reaction of phenols and enols with ferric ions
Phenols form a colourful complex with ferric ions whose colour depends
on the substituent of the aromatic ring.
H
ArO
6 ArOH

H
ArO

Fe3

OAr
Fe

ArO

OAr

+ 3H

OAr
H

Studied compounds: propane-2-ol, glycerol, phenol, 2-naphthol (-naphthol),

pentane-2,4-dione (acetoacetone), ethyl acetoacetate.

Propan

Glycer

Phen

2-

79

pentan

ethyl

unkno

e-2-ol

ol

ol

Napht
ol

e-2,4dione

acetoacet
ate

wn

Structure
of alcohol
Observati
ons
Experiment: 1 ml water is added to 1drop or ~15 mg of studied compound
in a test tube. Then one drop of 2.5% ferric chloride is added to this and the
changes are monitored. 1 ml bromine in water is added to the ferric chloride
solution of ethyl acetoacetate, and shaked in order to disrupt the enol-Fe(III)
complex by bromination of the double bond (the yellow colour of the
aqueous ferric chloride reappears).
Remarks: Enols or enol tautomeric forms react in a similar way with ferric
chloride as phenols. Bromine can ruin the complex by brominating the enol
form. Pentane-2,4-dione does not give a positive test, since in its enol form,
the OH is hydrogen-bonded to the carbonyl group. 2-Naphthol does not give
a positive either, since the resultant complex would be sterically hindered.

Preparation of cyclohexanone
OH

O
HOCl

C6H12O
Mol. Wt.: 100.16
m.p.: 24 C
b.p.: 161 C

2,4-DPH

C6H10O
Mol. Wt.: 98.14
b.p.: 157 C

HN
N
O2N

NO2

C12H14N4O4
Mol. Wt.: 278.26
m.p.: 162 C

Mix 0.2 cm3 cyclohexanol and 0.5 cm3 acetic acid in a test tube. Add
dropwise 5 cm3 of sodium hypochlorite to the reaction mixture within 10
minutes, and then wait for 20 minutes. Subsequently add 2 cm3
dichloromethane to the reaction mixture and shake it. Separate the organic
layer with Pasteur pipette and isolate the cyclohexanone as 2,4dinitrophenylhydrazone derivative. Add 4 cm3 2,4-dinitrophenylhydrazine
reagent to the organic layer and filter off the precipitation. Wash with water
and dry it. Check the purity of the product by TLC (toluene : ethyl acetate =
4:1) and the melting point measurement.
80

Preparation of 1,3-Dinitrobenzene (microscale)

(Electrophilic aromatic substitution, nitration)
NO2

HNO3
H2SO4
NO2
C6H6
Mol. Wt.: 78,11
b.p.: 80 C

C6H4N2O4
Mol. Wt.: 168,11
m.p.: 89 C

In a 100 ml Erlenmeyer flask, 2 ml cc. sulfuric acid and 2 ml cc. nitric

acid are stirred carefully while wearing protective google and gloves. Then
the flask is put in cold water bath in order to cool it down. 0.5 ml benzene is
measured into a test tube and added by a Pasteur pipette in three aliquots to
the prepared nitrating acid. During the addition of benzene (5 min), the flask
is shaded and cooled with water bath. As the addition of benzene is
completed, the reaction mixture is heated in a water bath at 100 C for ten
minutes. Then it is allowed to cool down and poured onto approximately 30
ml smashed ice. The formed yellow precipitate is filtered with vacuum and
washed with water twice. The purity of the product is checked by thin layer
chromatography and melting point measurement.

Preparation of 3-nitroaniline by the partial reduction of 1,3dinitrobenzene

NO2
Na2S

NO2

(X-1)S H2O

xS NaOH
NH2

NO2

In a three-neck flask equipped with mechanic stirrer, condenser and

dropping funnel, 2.0 g 1,3-dinitrobenzene is suspended in 100 ml water. Then
the stirring is started and the mixture is boiled. The solution of 3.0 g
crystalline potassium sulfide in 10 ml water is put in the dropping funnel. To
the boiling reaction mixture, 0.8 g sulfur and the potassium sulfide solution
are added successively in four portions. Sulfur is added through a funnel for
solid by removing the dropping funnel, and then after the replacement of the
dropping funnel, the solution is also added. Between each portion 5 minutes
81

interval is held. After completing the addition, the reaction mixture is stirred
and boiled for additional 20-30 minutes. Meanwhile the equipment for
vacuum filtration is to be set. As the reaction is completed the solid sulfur is
removed by filtering the hot solution. The crude product is crystallizing from
the filtrate upon cooling. The crystals are separated by filtration, washed with
some cold water and recrystallized without further drying. The weight of the
recrystallized product should be around 1.5 g. The purity of the crude product
and the recrystallized product is checked by TLC (eluent: toluene/ethyl
acetate 1:4) and melting point measurement.

Identification of amino derivatives of hydrocarbons.

1. The Hinsberg Test
Primary and secondary amines react with sulfonyl chlorides to form
sulfonamides:
R NH2 +

R'

H
N

O
Ar S Cl
O

O
Ar S NH N-substituted sulfonemide
O R

O
Ar S Cl
O

O R'
Ar S N
O R

N,N-disubstituted sulfonemide

Sulfonamide formation is the basis for a chemical test, called the

Hinsberg test that can be used to demonstrate whether an amine is primary,
secondary, or tertiary. A Hinsberg test involves two steps. First, a mixture
containing a small amount of the amine and benzenesulfonyl chloride is
shaken with excess potassium hydroxide. Next, after allowing time for a
reaction to take place, the mixture is acidified. Each type of amine primary, secondary, or tertiary - gives a different set of visible results
after each of these two stages of the test.
Primary amines react with benzenesulfonyl chloride to form Nsubstituted benzenesulfonamides. These, in turn, undergo acid-base
reactions with the excess potassium hydroxide to form water-soluble
potassium salts. (These reactions take place because the hydrogen attached
to nitrogen is made acidic by the strongly electron-withdrawing -SO2group.) At this stage our test tube contains a clear solution. Acidification
of this solution will, in the next stage, cause the water-insoluble Nsubstituted sulfonamide to precipitate:

82

R NH2 +

O
Ar S NH
O R

O
Ar S Cl
O

NaOH

water insoluble

O
Ar S N
O R

Na

O
Ar S NH
O R

HCl

water insoluble

water soluble

Secondary amines react with benzenesulfonyl chloride in aqueous

potassium hydroxide to form insoluble N,N-disubstituted sulfonamides
that precipitate after the first stage. N,N-Disubstituted sulfonamides do not
dissolve in aqueous potassium hydroxide because they do not have an
acidic hydrogen. Acidification of the mixture obtained from a secondary
amine produces no visible result - the nonbasic N,N-disubstituted
sulfonamide remains as a precipitate and no new precipitate forms:
R'

H
N

O R'
N,N-disubstituted sulfonemide
Ar S N
O R
water insoluble

O
Ar S Cl
O

If the amine is a tertiary amine and if it is water insoluble, no apparent

change will take place in the mixture as we shake it with benzenesulfonyl
chloride and aqueous KOH. When we acidify the mixture, the tertiary
amine dissolves because it forms a water-soluble salt.
R''
NH

Cl R'
R
water soluble

HCl
R'

R''
N

O
Ar S Cl
O
R

no reaction

Studied compounds: aniline, diethylamine, triethylamine

Procedure: To 1.5 ml 10% NaOH solution, two drops or ~30 mg studied
compounds and twodrops of benzenesulfonyl chloride are added in a test
tube and shaked for 10 minutes. If no change is observed the mixture is
slightly heated for 3-5 minutes and it is allowed to stand. If there is no
precipitate 0.5 ml aliquot of the sample is acidified with 10% HCl
solution.
If precipitate formation is observed from the basic solution, 1ml water is
added. If the precipitate does not dissolve the basic solution is removed
and 1ml 10% HCl solution is added.
Observations
Alkaline solution Acidic solution
Aniline
Diethylamine
Triethylamine
83

Unknown
Remark: For primary amines, the test is reliable for up to seven carbon
atoms, since with longer carbon chain the solubility of the sulfonamide
decreases.

2. Reactions of amines with nitrous acid

Nitrous acid (HONO) is a weak, unstable acid. It is always prepared
in situ, usually by treating sodium nitrite (NaNO 2 ) with an aqueous
solution of a strong acid. Nitrous acid reacts with all classes of amines.
The products that we obtain from these reactions depend on whether the
amine is primary, secondary, or tertiary and whether the amine is
aliphatic or aromatic.
Reactions of Primary Aliphatic Amines with Nitrous Acid
Primary aliphatic amines react with nitrous acid through a
reaction called diazotization to yield highly unstable aliphatic diazonium
salts. Even at low temperatures, aliphatic dia-zonium salts decompose
spontaneously by losing nitrogen to form carbocations. The carbocations
go on to produce mixtures of alkenes, alcohols, and alkyl halides by
removal of a proton, reaction with H 20, and reaction with X -:
Diazotizations of primary aliphatic amines are of little synthetic
importance because they yield such a complex mixture of products
RNH2 + NaNO2 + 2HX

0 - 5 C

R N N X

+ NaX + 2H2O

Aliphatic diazonium salt

(highly anstable)
-N2
Alkenes, alcohols, alkylhalides

R + X

Reactions of Primary Arylamines with Nitrous Acid

Primary arylamines react with nitrous acid to give arenediazonium
salts. Even though arenediazonium salts are unstable, they are still far more
stable than aliphatic diazonium salts; they do not decompose at an apprecia-

84

ble rate in solution when the temperature of the reaction mixture is kept
below 5C:
ArNH2 + NaNO2 + 2HX

0 - 5 C

Ar N N X

+ NaX + 2H2O

Arenediazonium salt
(stable if kept below 5 C)

Diazotization reactions of primary arylamines are of considerable

synthetic importance because the diazonium group (NN), can be replaced
by a variety of other functional groups.
Reactions of Secondary Amines with Nitrous Acid
Secondary aminesboth aryl and alkylreact with nitrous acid to
yield N-nitrosoamines. N-Nitrosoamines usually separate from the reaction
mixture as oily yellow liquids.
(CH3)2NH + HCl + NaNO2

H
N

CH3

(H3C)2N N O

N
N

+ HCl + NaNO2

N-nitrosodimethylamine

O
CH3 N-nitroso-N-methylaniline

Reactions of Tertiary Amines with Nitrous Acid

When a tertiary aliphatic amine is mixed with
nitrous acid, an equilibrium is established among the
tertiary amine, its salt, and an N-nitrosoammonium
compound.
While N-nitrosoammonium compounds are stable at low
temperatures, at higher term-peratures and in aqueous acid they
decompose to produce aldehydes or ketones. These re-actions are of little
synthetic importance, however
2 R3N + HX + NaNO2

R3NH X + R3N N O

Tertiary arylamines react with nitrous acid to form C-nitroso

aromatic compounds. Nitrosation takes place almost exclusively at the
para position if it is open and, if not, at the ortho position. The reaction
is another example of electrophilic aromatic substitution.
H3C
N
H3C

+ HCl + NaNO2

85

H3C
N
H3C

O
N

Observations
Buthylamine
Aniline
Triethylamine
Diethylamine
Nitrobenzene
Unknown
Experiment: Five drops of the studied compounds are disolved in cc HCl
solution (0.5 cm3) in a test tube and then cool down in an ice bath. Add three
drops of freshly cooled NaNO2 solution and wait for 2-3 minutes. If do not
observe neither gas formation nor precipitation add two drops of the solution
of 2-naphtol to it. Record the observations.
3. The Rimini reaction of aliphatic primary amines
The Schiff base formed from an aliphatic primary amine and acetone
gives colourful complex in the presence of sodium pentaciano-nitrosoferrate
(Na2[Fe(CN)5(NO)].
O
RNH2

H3C

NHR
CH3

H3C

NA2[Fe(CN)5NO]

CH3

colourful complex

Schiff base

Observations
Buthylamine
Aniline
Piperidine
Triethylamine
Nitrobenzene
Unknown
Experiment: One drop of the studied compound is dissolved in 3 ml water
and 1 ml acetone, and then one drop of freshly prepared (Na2[Fe(CN)5(NO)]
solution is added.
4. Reaction of aniline derivatives with bromine

86

Similarly to phenols, aniline derivative and N-alkylated aniline

derivatives are strongly activated for electrophilic aromatic substitution and
hence they can be readily brominated by aqueous bromine solution.

Observations
Cyclohexylamine
Aniline
N,N-dimethylaniline
Unknown
Experiment: Two drops of the studued compound are dissolved in 2 ml
water and aqueous bromine solution is added in a hood with continuous
shaking.
5. Complex formation of amine with Cu(II) ions.
Aliphatic amines form characteristic blue tetraamine complex with Cu(II)
ions, while the complexes formed with aromatic or secondary amines
precipitates due to their poor solubility.
Observations
Buthylamine
Aniline
Piperidine
Triethylamine
Unknown
Experiment: In a test tube, one drop of the studied compound is added to 0.5
ml water, then one drop of 10% CuSO4 solution is added and the mixture is
shaked. If there is no precipitate formation, more studied compound is added
dropwise while shaking the test tube.
Remark: The test is not specific. Other compounds like poliols, phenols,
enols also react with Cu(II) ions.

Chemical tests for oxo compounds

1. Chemical test for the identification of oxo compounds
Aldehydes and ketones give colourful insoluble 2,4-dinitrophenylhydrazones with 2,4-dinitro-phenylhydrazyne:
87

O
R

O2N

NH2
NH

O2N

N
H

NO2

O2N

NH2
NH

O2N

R
N

NO2
yellow/orange crystals

aldehyde

R
N
H

NO2

R
N

NO2
yellow/orange crystals

ketone

Experiment: To 8 drops of 2,4-dinitro-phenylhydrazyne reagent in test tube,

1 drop of or a few crystals of the studied compound are added. Formation of
coloured precipitate or crystals implies positive test.
formaldehyde benzaldehyde acetophenone

unknown

Structure of
oxo
compounds
Observatoins
Structure of
hydrazones

2. Oxidation of aldehydes by neutral potassium permanganate solution.

In contrast to ketones, aldehydes can be readily oxidized neutral
potassium permanganate solution.
O
R
H
aldehyde

KMnO4
R

OH

+ MnO2

KMnO4

no reaction

R
R
ketone

Experiment: In a test tube, 5 dropps or ~30 mg studied compound is

dissolved in 0.5 ml N,N-dimethylformamide and 1ml 1% potassium
permanganate is added and the mixture is shaked.
formaldehyd benzaldehyd

88

aceton

acetophenon

unknow

Structure of
oxo
compounds
Observation
s
3.Oxidation of oxo compounds by Jones reagent.
With Jones reagent (CrO3 and cc H2SO4 in acetone), the aldehydes are
oxidized much faster to the corresponding carboxylic acids than the ketones
and the aliphatic and aromatic aldehydes also show different reactivity.
O
R
H
aldehyde

CrO3
R

OH

CrO3

R
R
ketone

mixture of carboxylic acid

Experiment: In a test tube, two drops or ~15 mg of the studied compound is

dissolved in 0.5 ml N,N-dimethylformamide and 1 drop of Jones reagent is
added. The mixture is shaked and allowed to stand for for a few minutes. If
there is no change after a few minutes the test tube should be slightly heated.
formalde
hyde

propionalde
hyde

benzalde
hyde

aceto
ne

acetophe
none

unkno
wn

Structure
of oxo
compoun
ds
Observat
ions

4.Reaction of aldehydes with Tollens reagent

The ease with which aldehydes undergo oxidation provides a useful
test that differentiates aldehydes from most ketones. Mixing aqueous silver
nitrate with aqueous ammonia produces a solution known as Tollens' reagent.
The reagent contains the diamminosilver(I) ion, Ag(NH3)2+. Although this
89

ion is a very weak oxidizing agent, it oxidizes aldehydes to carboxylate

anions.

Ag

R'

Ag(NH3)2
O

OH

Experiment: In a test tube, to 1 ml of Tollens reagent, 1 drop of the studied

compound is added and while shaking the reaction is monitored for 5
minutes. If no change is observed the test tube is slightly heated in a 60 C
water bath for 3 minutes.
formalde
hyde

propionalde
hyde

benzalde
hyde

aceto
ne

acetophe
none

unkno
wn

Structure
of oxo
compoun
ds
Observat
ions

5. Iodoform test of methyl ketones

When methyl ketones react with halogens in the presence of base,
multiple halogenations always occur at the carbon of the methyl group.
Multiple halogenations occur because introduction of the first halogen (owing
to its electronegativity) makes the remaining hydrogens on the methyl
carbon more acidic:
When iodine is the halogen component, the bright yellow solid iodoform
(CHI3) results. This version is the basis of a laboratory classification test for
methyl ketones and methyl secondary alcohols (which are oxidized to methyl
ketones first under the reaction conditions):
O

O
CH3

+ 3I2

+ 3 OH

CI3

+ 3 I + 3 H2O

+ OH

CI3

90

R' +

CHI3
yellow crystals

Ag

Experiment: In a test tube, to two drops of the studied compounds 6 drops of

10 % NaOH sollution is added. The test tube is put in a 60 C water bath for
half a minute and KI-I2 solution is added dropwise until the brown colour
remains. The pH is checked and if it is acidic 5-6 drops 10 % NaOH solition
is added, heated.
formaldehyde

ethanol

acetone acetophenone

unknown

Structure of
oxo
compounds
Observations

Preparation of iodoform
O

I2

CHI3

NaOH
C3H6O
Mol. Wt.: 58.08
b.p.: 56-57 C

CH3COONa

Mol. Wt.: 393.73

o.p.: 119-123C

Add 1.0 g iodine and 0.5 cm3 acetone to the test tube. Place the test
tube to the ice bath and add dropwise 5 cm3 10 % solution of sodium
hydroxide while shake the test tube. The product precipitated as yellow
crystals. Filter off the product, wash with water and then with ethanol. Dry it
on air. Check the purity by melting piont measurement.

Preparation of benztriazol
(Diazotation and intramolecular ring closure of the diazonium salt)
NH2
NH2

NaNO2
CH3COOH
0-5 C

N+

NH2

C6H8N2
Mol. Wt.: 108.14
m.p.: 102 C

O
C CH3
O

- CH3COOH

N
N
N
H
C6H5N3
Mol. Wt.: 119.12
m.p.: 99-101 C

91

0.54 g 1,2-phenylenediamine is dissolved in the mixture of 1.2 ml

acetic acid and 2.5 ml water in a test tube. The test tube containing the
resultant solution is placed in ice-water bath. When the mixture is cooled
down, the precooled solution of 0.35 g sodium nitrite and 1 ml water is added
in small portions under continuous shaking and cooling. It is allowed to stand
for 5 minutes and then 25 % NaOH solution is added until it is neutralized
while still keeping the test tube in the ice-water bath (the pH is checked with
universal indicator paper). The precipitated substane is filtered in vacuum,
washed with water and dried. The purity of the product is checked by TLC
(eluent: chloroform/methanol 2:8) and melting point measurement.

Preparation of benzoic acid

O
COOH
1) NaOCl
2) HCl
C7H6O2
Mol. Wt.: 122.12
m.p.: 122.5 C

C8H8O
Mol. Wt.: 120.15
b.p.: 202 C

In a test tube, 0.2 ml acetophenone and 7 ml 5% sodium hypochlorite

is mixed (acetophenone is situated above the aqueous layer). Then the
mixture is placed in 50 C water bath and it is allowed to stand for 30
minutes while shaked and stirred occasionally. Formation of chloroform at
the bottom of the test tube indicates the completion of the reaction. The
reaction mixture is allowed to cool down and the chloroform layer is
removed by a Pasteur pipette. The sodium salt of the prepared benzoic acid
remains in the aqueous layer. While cooling, the solution is acidified with 10
% HCl, and the precipitated white crystals were filtered in vacuum, washed
twice with 3 ml cold water and air-dried. The purity of the product is checked
by TLC (eluent: hexane/acetone 7/3).

Preparation of 2,3-Diphenylquinoxalin

92

NH2

C6H5

C6H5

C2H5OH

+
NH2
C6H8N2
Mol. Wt.: 108.14
m.p.: 102 C

C14H10O2
Mol. Wt.: 210.23
m.P.: 95 C

C6H5

C6H5

C20H14N2
Mol. Wt.: 282.34
m.p.: 123-124 C

In 5 ml round-bottom flask equipped with a condenser, 0.25 g 1,2diphenylethane-1,2-dione (benzil or dibenzoyl) dissolved in 1 ml ethanol. To
the hot solution, 0.14 g o-phenylendiamin dissolved in 1 ml ethanol is added
and the mixture is boiled at 100 C on sand bath for 30 minutes. Then it is
allowed to cool to room temperature and the precipitate is filtered off, washed
with some ethanol (1-2 ml) and dried. Its purity is checked by TLC
(chloroform/methanol 9:1).

Preparation of 2,6-dibenzylidene-cyclohexanone
O

O
PhCHO
NaOH

C20H18O
Mol. Wt.: 274.36
m.p.: 118 C

C6H10O
Mol. Wt.: 98.14
b.p.: 155 C

0,1 g cyclohexanone and 0,22 g benzaldehyde are dissolved in

ethanol (1 cm 3 ) and 0.1 cm 3 5 M solution of sodium-hydroxide is
added to it. Shake the test tube and wait for 1 hour. The product
precipitate from the solution, filter off, wash with the mixture of
ethanol and water (ratio: 1 to 1) and then with cool methanol. Dry it
and check the purity by TLC and melting point measurement.

Preparation of acetylsalycilic acid (esterification with anhydride)

93

COOH
OH

COOH

Ac2O
cc. H2SO4

O
O

C7H6O3
Mol. Wt.: 138.12
m.p.: 155-156 C

C9H8O4
Mol. Wt.: 180.16
m.p.: 135 C

1 g salycilic acid and 1.4 ml acetic acid anhydride is measured into 50

ml round flask and two drops of conc. H2SO4 is added. By using a condenser,
the mixture is kept at 60 C for 20 minutes. Then 15 ml water is added and it
is allowed to cool to room temperature. The obtained precipitate is filtered
and the crude product is recrystallized from acetic acid/water 1:1.

Isolation and saponification of trimyristin from nutmeg

The nutmegs Myristica are a genus of evergreen trees indigenous to

tropical southeast Asia and Australasia. They are important for two spices
derived from the fruit, nutmeg and mace.
Trimyristin is an ester with the chemical formula C45H86O6. It is a
saturated fat which is the triglyceride of myristic acid. Trimyristin is found
naturally in many vegetable fats and oils. In nutmeg it accounts for the sweet
smell and taste of the spice. Trimyristin is a white to yellowish-gray solid that
is insoluble in water, but soluble in ethanol, benzene, chloroform,
dichloromethane, and ether.
O

O
O
O

O
O

C45H86O6
Mol. Wt.: 723.16

The isolation of trimyristin from powdered nutmeg is a common

introductory-level college organic chemistry experiment. It is an
uncommonly simple natural product extraction because nutmeg oil generally
consists of over eighty percent trimyristin. Trimyristin makes up between 2025% of the overall mass of dried, ground nutmeg. Separation is generally

94

effected by steam distillation and purification uses extraction from ether

followed by distillation or rotary evaporation to remove the volatile solvent.
The extraction of trimyristin can also be done with diethyl ether at room
temperature, due to its high solubility in the ether.
Experiments: Powdered nutmeg (5 g) and ether (50 cm3) is stirred in
an Erlenmayer flask (250 cm3) for 10 minutes in a hood. Filter off the nutmeg
and evaporate the solvent using rotatory evaporator. Add 5-10 cm3 acetone to
the residue and put the solution to the freezer. The trimyristin is precipitated
from the solution. Filter off the white crystals, dry on air and measure the
mass of product, determine trimyristin contents of nutmeg.
Saponification of trimyristin: Add 10 cm3 6M aqueous solution of
sodium hydroxide to the trimiristine (0,3 g) in ethanol (10 cm3) in round
bottom flask. Take a condenser into the flask, and reflux by heating with
water bath for 60 minutes. Then pour onto the water (100 cm3) and add cc.
HCl (7 cm3) to it. Filter off the precipitation, wash with water. Control the
purity by TLC (benzene : ethanol = 9:1, detection: iodine).
O

O
O
O

O
O
C45H86O6
Mol. Wt.: 723.16
mp.: 55-56 C
NaOH

OH
OH

HO

C14H28O2
Mol. Wt.: 228.37
m.p.: 53-54 C

Preparation of N-benzoyl glycin

95

OH

O
O
H2N

COOH

C2H5NO2
Mol. Wt.: 75.07
m.p.: 245 C

Cl

H
N

COOH

NaOH

C7H5ClO
Mol. Wt.: 140.57
b.p.: C

NaCl

C9H9NO3
Mol. Wt.: 179.17
m.p.: 187 C

0.5 g glycin is dissolved in 5 ml 10% NaOH solution in a 25 ml

Erlenmeyer flask. Under stirring, 0.9 ml benzoyl chloride is added dropwise
and the mixture is allowed to stand for 20 minutes at room temperature while
it is stirred occasionally. Then the flask is placed in icy water and acidified
with cc. HCl. The obtained precipitate is filtered and washed with 33ml
water. The product is recrystallized from water.

96