Practical Organic Chemistry

A Laboratory Handbook

For B. Sc. (General) Degree

(2000 Level)

Department of Chemistry

University of Peradeniya

(2023)

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PRACTICAL ORGANIC CHEMISTRY
1. MELTING POINT DETERMINATION: IDENTITY AND PURITY OF ORGANIC
COMPOUNDS
Experiment I. Determination of the mps of crude and pure benzoic acid.
Experiment 2. Identification of an unknown compound using mixed mp
2. RECRYSTALLISATION
Experiment 3. Recrystallisation of impure acetanilide from water.
Experiment 4. Recrystallization of impure salicylic acid.
Experiment 5. Recrystallisation using a solvent pair.
3. DISTILLATION, SEPARATION AND PURIFICATION OF ORGANIC LIQUIDS
Experiment 6. Separation of two miscible liquids by simple and fractional distillation.
4. REFLUXING
Experiment 7. Hydrolysis of an amide.
Experiment 8. Hydrolysis of an Ester.
5. EXTRACTION: A TECHNIQUE FOR SEPARATION AND ISOLATION
Separation of Simple Binary Mixtures of Organic Compounds.
Experiment 9. Separation of an Acidic-Neutral Mixture
Experiment 10. Separation of a Base-Neutral Mixture
Experiment 11. Acetylation of an aromatic base.
Experiment 12. Benzoylation of an amine (Schotten-Baumann reaction)
Experiment 13. Separation of a mixture of chloroaniline (base), salicylic acid (acid) and
p-dichlorobenzene (neutral substance) into its components by extraction.
ISOLATION OF NATURAL PRODUCTS.
Experiment 14. Isolation of caffeine from tea.
Experiment 15. Extraction of eugenol from cloves (steam distillation).
Experiment 16. Extraction of Cumin oil from cumin seeds.
4. SYNTHETIC REACTIONS OF ORGANIC COmpOUNDS
Experiment 17. Synthesis of Aspirin
Experiment 18. Preparation of fumaric acid from maleic anhydride.
Experiment 19. Preparation of Oximes.
Experiment 20. Acetylation of an Amine or a Phenol.
Experiment 21. Benzoylation of an Amine or a Phenol
Experiment 22. Synthesis of β-naphthyl acetate.
Experiment 23. Oxidation of β-naphthol with iron(iii) chloride to give di-β-naphthol.
Experiment 24. Oxidation of aromatic compounds.
a. Preparation of anthraquinone from anthracene
b. Preparation of benzoic acid from benzyl chloride (hydrolysis and oxidation)

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 Experiment 25. Preparation of succinanilic acid
Experiment 26. Preparation of p-nitroaniline and its benzal (benzylidene) derivative
a. Acetylation of aniline (Expt. No. 20)
b. Nitration of acetanilide
c. Hydrolysis of p-nitroacetanilide
d. Preparation of benzal (benzylidene) derivative of p-nitroaniline
Experiment 27. Nitration of phenol
Experiment 28. Preparation of 1, 3, 4, 6-di-O-benylidene-D -mannitol.
Experiment 29. Preparation of a-penta-O-acetyl- D-glucose.
Experiment 30. Preparation of β-Penta-0-acetyl-D-glucose.
Experiment 31. Carbanion additions
a. Preparation of sorbic acid from crotonaldehyde (Knoevenagel reaction)
b. Preparaton of dibenzalacetone (Claisen-Schmidt reaction)
Experiment 32. Preparation of sulphanilamide
a, Preparation of p-acetamidobenzenesulfonamide
b. Preparation of p-aminobenzenesulfonamide (sulfanilamide).
Experiment 33. Grignard Reaction
a. Preparation of the Grignard reagent
b. Synthesis of triphenylmethanol
Experiment 34. Preparation of anthranilic acid (Hoffman degradation)
5. INDUSTRIAL APPLICATIONS
Experiment 35. Extraction of a Fat/Oil
Soaps and Detergents
Experiment 36. Preparation of Soap.
Properties of soap and a synthetic detergent.
Experiment 37. Preparation of Terephthalic Acid.
Preparation of "Terylene"
Dyes and Dyeing
Experiment 38. Direct dyeing of wool and silk
Experiment 39. Direct dying of cotton.
Mordant dyeing of cotton.
Diazotisation and Preparation of Azo Dyes
Experiment 40. Preparation of Benzene-azo-2-naphthol.

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PRACTICAL ORGANIC CHEMISTRY
Organic Chemistry is concerned with the preparation of compounds, and the study of their
properties and reactions. Many experiments are designed to prepare a specific compound, but
forming the compound is less than half the problem. The mixture that results contains your product
and much more. Separation, purification, analysis and identification are required to isolate the
desired products and to prove that you have made what you set out to do.
This laboratory course has been designed to introduce many of the basic experimental techniques
used for the preparation and purification of organic compounds. It should teach you good synthetic
methods and help you to develop an analytical approach to problem solving in the laboratory. At the
beginning of the course you will be introduced to simple techniques which are used repeatedly in
experimental organic chemistry and biochemistry. You will be given many opportunities to improve
your skills in these techniques.

Come to the laboratory with a mental plan, understanding what must be done. Read the laboratory
manual, lecture notes, relevant textbooks, and be ready to start at once on the practical work for that
day. Some students attempt to conduct an experiment by holding the flask/test tube in one hand
while keeping a finger of the other hand on the relevant place in the laboratory manual. This can only
lead to trouble! The experiment is slowed to such an extent that it is not possible to complete the
experiment during the time assigned to it. Study before coming to the laboratory so that you are able
to understand what is happening during an experiment and make maximum use of your laboratory
hours. The questions after each experiment have been included to help you to be well prepared.

Training in practical chemistry must include knowledge of safety measures necessary for handling
both chemicals and equipment. It is essential that you learn and understand these precautions
thoroughly. You are therefore advised to read the relevant sections on Laboratory Safety found in
any one of the practical texts given below.

1. Elementary Practical Organic Chemistry. Part A. I. Vogel, p. 45-168

2. Organic Experiments. L. F. Fieser and K. L. Williamson, Chapter 5-9.
3. Techniques and Experiments for Organic Chemistry. A. Aul, p. 29-128.
4. Practical Organic Chemistry. F. G. Mann and B. C. Saunders.
5. Experiments in Organic Chemistry. L. F. Fieser.
6. Organic Chemistry Laboratory. O. R. Rodig, C. F. Bell and A. K. Clark, Chapters 1-5.
7. Organic Chemistry Laboratory. O.R. Rodig, C. E. Bell and A. K. Clark

1. MELTING POINT DETERMINATION: IDENTITY AND PURITY OF ORGANIC COMPOUNDS

Most crystalline organic compounds have characteristic melting points (mp) that are sufficiently
low (50°-300°) to be determined with simple equipment. Organic chemists use mps to identify
crystalline compounds and to get an indication of their purity. A nickel spatula is used to grind a small
amount of the substance on a clean, dry section of a porous tile and finely powdered material is
packed into a thin walled capillary tube sealed at one end. The capillary tube is heated in a
well-stirred liquid bath, or in a metal block. The temperature at which the substance begins to liquefy
and the temperature at which it is completely liquefied give the mp range. A pure crystalline organic
compound has a mp range of 1-2 °C. Impurities (including residual solvent) lower the mp of a
compound and also widen the mp range. The mp range can be influenced also by the size of the
crystals, the amount of material, the density of its packing and the rate of heating of the bath.

Two different compounds, even with the same mp will be mutual impurities if mixed with each other
and would lower and widen the mp range. Hence mps are used to determine whether two substances

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are the same compound or whether they are different ones. Two crystalline compounds with the
same mp are identical if their mp is not lowered when mixed.

Experiment I. Determination of the mps of crude and pure benzoic acid.

Experiment 2. Identification of an unknown compound using mixed mp

Determine the mp of the unknown compound A. Consult Table 1 and make a preliminary
identification of A. Combine equal amounts of two compounds (grind them together on a porous
plate using a nickel spatula) then place the mixture in the mp capillary tube and determine the mixed
mp. Record your observations.

Table 1
Compound mp (°C) Compound mp (°C)
Biphenyl 70-71 Benzoic acid 121-122
Phenyl benzoate 69-70 2-naphthol 121-122
m-dinitrobenzene 90-91 Cinnamic acid 133-135
o-nitroacetanilide 92-93 Urea 133-135
Acetanilide 114-115 Salicylic acid 156-158
p-benzoquinone 114-115 Benzanilide 160-161

2. RECRYSTALLISATION

Crude solid compounds isolated from a chemical synthesis or from an extraction are most often
purified by a technique known as recrystallisation, based on differences in solubility of solids in a
given solvent system. Generally the solubility of a compound increases with temperature. Therefore,
the compound can be recovered in a solid form, when a saturated solution of the compound is
allowed to cool. Purification results because the soluble impurities are present in much smaller
concentration, and hence will remain in solution when the solution is cold. The purified crystals are
separated by a procedure called suction or vacuum filtration (Fig. 1a). Insoluble impurities can be
removed by filtration of the hot solution (Fig. 1b).

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 Fig 1a. Vacuum Filtration

The most important step in recrystallisation is to find the correct solvent. An ideal recrystallisation
solvent should have the following features.

● Dissolve the crude substance readily when hot but only to a small extent when cold.
● Dissolve the impurities readily so that they will remain in solution even when cooled, or
dissolve only to a small extent. If impurities do not dissolve at all they can be removed by hot
filtration or centrifugation.
● Does not react with the substance being recrystallised.
● Have a relatively low boiling point (bp) and volatilize readily so that it can be removed readily
from the pure compound.
● Bp is lower than the mp of the solid to be purified. A situation where the solid merely "oils
out", i.e. melts, but does not dissolve is avoided.

Sometimes one single solvent does not have all of the above requirements; in such cases a second
solvent, miscible with the first, is added to the latter to form a "solvent pair", in order to modify or
intensify the properties of the first solvent and make the system suitable for recrystallisation. In the
"solvent pair"; one solvent dissolves the compound well, while the other solvent dissolves the
compound poorly. To find a suitable solvent, add a little solvent to a small amount of the substance to
be purified and boil gently. (Use a water bath if the solvent is flammable). If the compound is slightly
soluble in the cold solvent but dissolves readily in hot solvent, cool the test tube and see whether
crystallization occurs.

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 Fig. 1b: Hot Filtration

Use of Animal Charcoal (Norit).

Finely divided charcoal can be used to remove coloured, soluble impurities. These coloured
substances attach to the activated surfaces of charcoal by adsorption, and since charcoal is not
soluble in any solvent, the adsorbed impurities can be filtered off. The decolorizing action of charcoal
works best in aqueous and some alcohol solvents, least well in hydrocarbon solvents. Charcoal is
added in small amounts to the saturated solution to which a slight excess of solvent has been added.
(The hot saturated solution must be cooled briefly before charcoal is added, since the latter causes
frothing when added to a boiling solution). The solution is gently boiled/refluxed for approximately 5
min while the flask is swirled continuously to avoid bumping. Charcoal is then removed by hot
filtration.

Method I
i. Dissolve the impure sample in the minimum volume of a suitable solvent at or near the bp of
the solvent, which should be removed readily from the pure compound. To obtain a good
recovery of purified material, large volumes of solvents must not be used. About 3-5% more
solvent than the minimum required is used so that the hot solution will not be saturated. This
helps to prevent separation of crystals and clogging of filter paper during filtration of the hot
solution.
ii. Filter the hot solution (if it contains insoluble impurities) to remove insoluble impurities . Use
a fluted filter paper and a stemless (or short stemmed) funnel which should be heated (pass
hot solvent through it) before filtration. The receiving container can also be heated to avoid
immeadiate solidification of the compound.
iii. Allow the solution to cool and crystals of the compound to spontaneously grow. If the solution
cools to room temperature and no crystals are formed, the solution may be super-saturated.
Here, the walls of the container may be gently scratched, or a crystal of the pure compound
introduced to act as seeds for the crystallization to begin.
iv. Filter the crystals under suction and separate the crystals from the solution (mother liquor).
v. Wash the crystals with a little cold solvent to remove mother liquor.
vi. Air dry the crystals by draining well under suction for a few minutes to remove the last traces
of solvent. Use a clean spatula and press the crystals as dry as possible on the filter. Stop the
suction by first pulling the tubing off at the water pump (to prevent back suction) then
shutting off the pump.

Method II
Mixed solvents for recrystallization.
If the compound is very soluble in one solvent and is almost insoluble in another solvent, a mixture of
the two solvents may be used for recrystallisation. The two solvents must be completely miscible.
Pairs of solvents which are used frequently are methanol (or ethanol) and water, CH2Cl2 and hexane
(or light petroleum). Ethanol, methanol and petroleum are inflammable (use a water bath and keep
fumes away from any flames).

Experiment 3. Recrystallisation of impure acetanilide from water.

The acetanilide provided contains some water insoluble impurities. Recrystallise and record the mp
of pure dry acetanilide. Submit your sample.

Experiment 4. Recrystallization of impure salicylic acid.

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Determine the mp of the impure salicylic acid. Check the solubility of salicylic acid in water and
ethanol. Which of these is the better solvent? Recrystallise the impure sample of salicylic acid, which
contains a soluble, coloured impurity. Submit your sample and record the mp of impure and pure
salicylic acid.

Experiment 5. Recrystallisation using a solvent pair.

m-Dinitrobenzene (or p-dibromobenznene) is soluble in cold ethanol. Hence ethanol is not a good
recrystallising solvent for these compounds. These compounds have a very low solubility in cold as
well as hot water. Since ethanol and water are miscible it is possible to find a mixture of the two
solvents in which the compounds are soluble when hot but relatively insoluble when cold.

Method. Dissolve the sample in the minimum volume of hot methylated spirit using a steam bath. Add
water drop by drop until the solution is slightly turbid. Warm until a clear solution is obtained and
allow to cool. If insoluble impurities are present the hot mixture should be filtered before water is
added. Submit your sample with a record of its mp.

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Flow Chart for Recrystallization

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 3. DISTILLATION, SEPARATION AND PURIFICATION OF ORGANIC LIQUIDS

Distillation is a process which involves heating a substance (to its bp) until it vaporizes, condensing
the resulting vapors by cooling and collecting the condensate (distillate) in a separate flask. (Fig. 2). In
the process called refluxing (Fig. 3) the condensate is returned to the original vessel. Distillation is the
most common method used to separate and purify organic liquids. The temperature remains constant
throughout the distillation of a pure liquid. Bp at a given pressure is a characteristic property of a
pure liquid. The distillation technique is useful for separating a mixture when the components of the
mixture have different bps and if the following conditions apply:

● At least one component in the mixture is volatile (non-volatile solids present would be left
behind in the distillation flask).
● If two or more volatile liquids are to be separated, then they must have widely different bps
(e.g., 50˚ or more)
● The components must not from an azeotrope (see below).

Azeotrope. Sometimes a mixture of two or more liquids distils at a constant temperature and with a
constant composition even though separately the components have different bps. This occurs when
intermolecular interactions (e.g., H-bonding) take place between the components of the mixture.
Such systems are called azeotropes and an example of one is ethanol and water, which form an
azeotropic mixture at 95% and 5% respectively.

Experiment 6. Separation of two miscible liquids by simple and fractional distillation.

The extent of the separation depends on the components of the mixture as well as the apparatus and
operating conditions.

Method. Assemble a simple distillation apparatus Fig 2a (distilling flask, water bath, thermometer,
water condenser, adapter and receiver) and Fig 2b (fractionating column added for fractional
distillation). Place a mixture of acetone (15 ml) and water (30 ml), add a few boiling stones (an inert
material with small pores that provide sites where bubbles can form, so that even boiling occurs and
"bumping" is prevented) and heat the flask in a water bath. Adjust the heating so that boiling takes
place steadily and drops fall at a rate of about one per second into the receiver. Record the
temperature when you collect 2, 4, 6..... 30 ml of distillate. Record your results in a chart as shown
below.
Plot the temperature against the volume distilled, on graph paper for both simple and fractional
distillation (use two different colours to distinguish between them). Comment on the efficiency of
each type of distillation for separation of the mixture.

Simple / Fractional Distillation

Volume of the Observed Volume of the Observed Volume of the Observed
Distillate / ml Temperature Distillate / ml Temperature Distillate / ml Temperature
2 12 22
4 14 24
6 16 26
8 18 28
10 20 30

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 Fig. 2a: Simple Distillation Fig. 2b: Fractional Distillation

Fig. 2c: Steam Distillation

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 4. REFLUXING

A substance can be heated for a long time without boiling away the solvent by attaching a condenser
to the flask containing the boiling solution. The solvent vapor condenses and returns as a liquid into
the boiling solution. The continuous boiling, vaporization and back flow of condensate is called
refluxing. Refluxing is used for many reactions, and also for recrystallising when the solution process
is slow, or when recrystallising large samples from volatile inflammable solvents.

Unlike the hydrolysis of acyl halides or anhydrides, hydrolysis of amides, esters and nitrites requires
boiling with aqueous acid or base. Esters are volatile and refluxing is necessary also because most
esters are volatile and may vaporize before they are hydrolyzed.

Fig. 3a: Reflux Apparatus

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 Fig. 3b: Reflux with addition

Experiment 7. Hydrolysis of an amide.

Method. Place benzamide (1 g) and aq. 10% NaOH solution (15 ml) and some boiling stones in a 100
ml round bottomed (rb) flask fitted with a reflux water condenser and boil gently for 30 min. Test the
gas evolved with litmus. Cool the solution in ice water and add conc. HCI until strongly acidic, allow
the mixture to stand in ice water for a few minutes, filter at the pump, wash with ice cold water and
drain well. Recrystallise from hot water.

1. Determine the mp of the pure dry acid. Record the literature mp value (giving literature
source) for benzoic acid.
2. Calculate the % yield of benzoic acid.
3. What is the gas evolved during hydrolysis of benzamide?
4. Outline a mechanism for the hydrolysis of benzamide to benzoate ion in 10% NaOH and
indicate the nucleophile.
5. Why was conc. HCI added at the end of the reaction?

Experiment 8. Hydrolysis of an Ester.

Method. Follow the procedure described above using an ester (1 ml) and boil gently with aq. 10%
NaOH (15 ml) until the ester layer disappears (20 – 30 min) Cool the solution in ice water and
cautiously acidify with conc. HCI. Allow the mixture to stand in ice water for 10 min, filter at the
pump, wash with ice cold water and drain well. Recrystallise the product from water.
1. Determine the mp of the pure dry acid. Record the literature mp value (giving literature

13
 source) for benzoic acid.
2. Use the density (or specific gravity) of the ester and calculate the % yield of benzoic acid
obtained from your reaction.
3. Outline a mechanism for hydrolysis of an ester using i. 10% NaOH and ii. dil. sulphuric acid.
Indicate the nucleophile in each reaction.

5. EXTRACTION: A TECHNIQUE FOR SEPARATION AND ISOLATION

The product of a reaction must be separated from by-products, excess reactants, impurities and other
substances that are present in the reaction mixture. Similarly, a substance from a natural source is
always mixed with other substances. Extraction is a common technique used to separate a desired
organic product from a reaction mixture, or to isolate an organic substance from its natural source.

Liquid-liquid extraction is widely used for separating mixtures. The components of the mixture
can be either liquids or solids. The method is based on the difference in solubility of the components
in two immiscible solvents. An organic and an aqueous solvent are sometimes used. Inorganic
substances dissolve in the aqueous phase and can be separated from organic compounds which are
soluble in the organic solvent. In general, several extractions using a small volume of solvent are
more effective than a single extraction using a large volume of solvent.

The mixture is dissolved or suspended in the first solvent and then this solution is introduced
second solvent. Even though it is much more soluble in the second solvent than in the first, it still has
the ability to dissolve in the first solvent. The extent of this ability is called the partition coefficient of
extraction. The extraction step is repeated 2-3 times, each time removing the extracting medium (this
would be the second solvent), and then adding a fresh portion of this solvent to the separating funnel
containing the original solution of the mixture. Successive extractions are done to ensure the best
possible separation of the components in the mixture. A pictorial description of the results of
successive extraction is given in Fig.4. In general, several extractions using a small volume of solvent
are

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Fig. 4: Separation by succesive extractions.

15
more effective than a single extraction using a large volume of solvent. The solvent selected for
extraction should;
● Readily dissolve the substance to be extracted.
● Be sparingly soluble in the solvent from which the substance is to be extracted.

Emulsions
Often the two solvents in an extraction do not separate cleanly; and an emulsion (foamy layer)
forms between them. An emulsion is a finely dispersed suspension of one liquid layer in another.
Separation is not possible until the layers become distinct again. It is possible to get rid of an emulsion
by just waiting for 10 —20 min and swirling the flask gently. If water is one of the solvents adding
some saturated NaCl solution may break up an emulsion. Saturated NaCl is much less soluble in the
organic solvent than water and hence will pull the water out of the organic layer.

Method
● When using a separating funnel, always keep it upright, supported by a clamp (Fig 5b). Do not
lay it on its side, either on the bench or in the sink.
● Pour the solution to be extracted into the separating funnel (make sure the stopcock is closed
and a beaker is under the funnel). Solutions should never occupy more than 2/3 of the volume
of the funnel. The stopper should rest on a clean watch glass until just before use. Do not keep
a closed separating funnel on the ring clamp.
● Stopper the funnel, hold it inverted making sure that the fingers of hand hold on to the closed
stopcock and the forefinger of the other hand holds onto the stopper (Fig. 5a). Point the stem
away from yourself (and your neighbours), gently shake the funnel, and then open the
stopcock carefully to release the initial pressure. Close the stopcock, shake more vigorously
this time and release the pressure again. This process is repeated until no more pressure
develops. The funnel is returned to the ring clamp, the stopper is removed immediately and
the layers allowed to separate.
● After the two layers have cleanly separated, drain the heavier bottom layer through the
stopcock. The top layer can then be removed from the funnel by decanting it through the top
of the funnel.

Drying procedure
After an extraction, the organic layer is usually saturated with water, and the latter must be removed
before the organic solvent is removed. Drying agents, such as inorganic salts that can form hydrates
are used to absorb this small amount of water. Commonly used drying agents are anhydrous
magnesium sulfate, sodium sulfate, and calcium chloride (for hydrocarbons or alkyl halides).
For solutions, knowing the amount of drying agent to add is a matter of acquiring experience. In
general, the amount of drying agent added should not exceed one tenth of the liquid volume. On
swirling the corked flask, if all of the drying agent remains clumped at the bottom and does not swirl
freely, not enough of the agent has been added. But as long as some of the agent moves freely as
individual grains, then you have probably added enough. However, it might be wise to check this
swirling again in 5 minutes. The solution is allowed to dry over the salt for at least 10 minutes, with
occasional swirling. The salt is removed by gravity filtration through a fluted paper (remembering you
have just removed water from this solution, so any further glassware that this solution will come in
contact with must be totally dry).

Solvent removal
The usual methods of removing an organic solvent from the desired compound are:
1. Evaporation on a steam or hot water bath (this must be done in a fume cupboard)
2. Evaporation at reduced pressure
3. Distillation (if the solute is a high boiling liquid or solid, see Fig. 6a)
4. Evaporation on a rotary evaporator- (see Fig. 6b).

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Easily reversed chemical reactions can be used to effect separations by extraction. e.g., dil. NaOH
converts organic acids into their sodium salts. A water insoluble acid can be converted into its water
soluble sodium salt.
RCO2H + NaOH → RCO2-Na+ + H2O
A mixture of a neutral and an acidic water insoluble compound can he separated by shaking
with Na0H when the acid as its sodium salt will dissolve in the aqueous layer whereas the neutral
compound will remain in the organic layer. The organic layer can be separated and the aqueous layer
acidified using a strong acid. An acidic substance can therefore be easily separated from neutral or
basic contaminants by extraction with aqueous alkali.
RCO2-Na+ + H+ → RCO2H + Na+ CI-

Fig. 5a: Use of Separatory Funnel Fig. 5b: Separating layers in Funnel

Phenols similarly undergo salt formation with alkali, but are usually less acidic than carboxylic acids.
ArOH + Na+OH- → ArO-Na+ + H2O

Aqueous NaHCO3 converts carboxylic acids to sodium salts but is not alkaline enough to
convert phenols to their salts. Thus NaHCO3 can be used to separate a mixture containing a
carboxylic acid and a phenol.
Dilute aqueous acids can be used to extract basic compounds, particularly amines, from
neutral and acidic substances by converting them to water soluble alkylammonium salts. Alkaloids,
which are nitrogenous bases found in plants, are usually extracted from plant sources by this method.
RNH2 + H-Cl+ → RNH3+ Cl-

After separation of the organic and aqueous layers, the amine can be recovered from the
aqueous layer by making the solution alkaline.
RNH3+Cl- + Na+OH- → RNH2+ H2O + Na+Cl-

Solvents
In the following experiments, the two-phase systems usually consist of an aqueous phase and an
immiscible organic phase, the compound to be isolated being extracted into the organic phase. The
organic solvent can be diethyl ether, chloroform or petroleum ether, etc. The total volume of solvent
placed in a separating funnel should not exceed 2/3 of the total funnel volume, otherwise proper

17
mixing is difficult. Dissolve your original mixture in a minimum amount of the first solvent. For the
second solvent (the "extracting medium"), it is better to divide the given volume of extraction solvent,
and do several extractions. This gives a better result than using the entire amount for one extraction.
The total volume of the solvent used for the successive extractions usually approximates the total
volume of the other solvent e.g., 45 ml of an aqueous solution is extracted with three 15 ml portions
of diethyl ether (NOTE: ether is flammable and must be removed carefully. A rotary evaporator may
also be used, See Fig. 6).

Separation of Simple Binary Mixtures of Organic Compounds.

Carry out a small-scale test and find out the solubility of the mixture in i. acid and ii. base and
determine whether it is a neutral-acid or neutral-base mixture of organic compounds. For a
neutral-acid mixture, the acidic component is converted by NaOH into its soluble sodium salt while
the neutral substance remains undissolved. A neutral-base mixture is treated with dil. HCl, which
dissolves the basic component, leaving the neutral component undissolved. The procedures outlined
below apply only to those mixtures in which the neutral substance is insoluble in water.

Experiment 9. Separation of an Acidic-Neutral Mixture

Method. Add aqueous 5% NaOH (15 ml) to the mixture (1.5 g) and shake well in a conical flask. Check
with litmus to ensure that there is excess of base. Filter under suction and retain the filtrate for
isolation of the acidic compound. Wash the solid neutral compound C on the filter with a little 5%
NaOH, wash with water and drain well by suction. Recrystallise from a methanol, record its mp and
submit your sample of C. Cool the filtrate and add dil. HCl gradually until it is acidic to litmus. If a
precipitate is obtained, filter at the pump, wash with cold water and drain well. Recrystallise from
water, use 0.2 g of this sample for the preparation of the benzylthiouronium salt (see below), and
submit the remainder of the pure dry carboxylic acid D. If a precipitate is not obtained, use a
separating funnel to extract the solution with CH2Cl2. Dry the organic layer with anhydrous Na2SO4

18
and distill off the solvent. The remainder in the flask is the acidic component D. Determine the
melting point of the carboxylic acid D.

Preparation of benzylthiouronium salts. Dissolve 0.2 g of the carboxylic acid D in a minimum of hot
water (about 50 ml) and add 5% aqueous sodium hydroxide until the solution is just alkaline to
methyl-orange, then add 1 drop of dilute hydrochloric acid. Pour this solution of the sodium salt into
solution of 0.4 g of benzylthiouronium chloride, [C6H5CH2SC(NH2)NH2]+Cl-, in 2 ml of water, and cool
the stirred mixture in ice-water. Filter off the benzylthiouronium salt which has seperated, and
recrystallise from ethanol containing 10% of water. Determine the melting point of the
benzythiouronium salt of D.

Experiment 10. Separation of a (Solvated) Base-Neutral Mixture

Method
Measure 10 mL of the base neutral mixture (dissolved in CH2Cl2 ), and place in a 100 mL separatory
funnel. Dilute 25 mL of dil. HCl to 50 mL with distilled water, and add 15 mL of the diluted acid to the
separatory funnel. Shake well, to allow the organic base to react with the acid and extract into the
water phase. Separate the aqueous and organic layers, and repeat the extraction using 15 mL x 2 of
dilute acid solution. Check the organic layer with red litmus paper, if it turns blue, carry out another
extrac tion.
Combine the (three ) aqueous extractions, and add a base until the solution is basic to litmus, and no
further precipitate is formed . F ilter under suction , and recrystallize the crude basic compound using
water.
Evaporate the CH2Cl2 in the organic extract and recrystallize the neutral compound using methanol.
Carry out the carbylamine test on the basic component .( Remember t o hydrolyse any carbylamine
formed by cooling and adding conc . HCl at the end of the test. What happens to the carbylamines
when HCl is added?). W hich functional group is indicated by a positive carbylamine reaction.

19
ISOLATION OF NATURAL PRODUCTS.

Experiment 16. Extraction of Cumin oil from cumin seeds.

The major volatile constituent of the seeds of cumin (Cuminum cyminum) is

p-isopropylbenzaldehyde (cuminaldehyde). Cumin oil is steam distilled and cuminaldehyde is isolated
as the semicarbazone derivative.
Method. Place cumin seeds (75 g) and distilled water (250 ml) in a 500 ml round bottomed flask and
steam distil the mixture with a 250 ml conical flask as a receiver. Steam distil until no more oil is
observed in the condensate (100-150 ml of distillate). Extract the distillate with CH2Cl2 (25 ml x 2).
Dry the organic extract over anhydrous Na2SO4 and remove the CH2Cl2 by distillation. Transfer the
oily residue with a little solvent into a 50 ml conical flask. Remove the solvent on a steam bath and
prepare the semicarbazone of cuminaldehyde as follows:
Dissolve semicarbazide hydrochloride (0.1 g) and anhydrous NaOAc (0.15 g) in water (1 ml) and add
ethanol (1.5 ml). Add this solution to the cumin oil, warm briefly on a steam bath, cool and allow to
crystallise from ethanol and drain well. Record the mp of the dry semicarbazone and submit your
product.
1. Why was NaOAc added for semicarbazone formation?
2. Outline a mechanism for semicarbazone formation.

4. SYNTHETIC REACTIONS OF ORGANIC COmpOUNDS

Experiment 17. Synthesis of Aspirin

Salicylic acid has been used in medicine for many years, either as the sodium salt or as an ester.
Salicylates are antipyretics, i.e., they lower the body temperature in a person who has a fever.
Salicylates are also mild analgesics, which relieve certain types of pain (headache, rheumatism).
Sodium salicylate irritates the lining of the stomach and this is why various esters are now used in
place of the free acid or salt. Aspirin is the most common salicylate used in medicine today. It is the
sodium salt of acetylsalicylic acid.
Method. Add acetic anhydride (1.0 ml) to salicylic acid (0.5 g) contained in a dry boiling tube. Add 2-3
drops of conc. H2SO4, mix well, cork the tube lightly and warm on a boiling water bath (100°C) for 15
min. Remove the tube from the water bath and carefully add ice water (5 ml) ( cool in ice bath before
adding the ice water). Stir and cool the tube in an ice bath. Filter the product at the pump and drain
well. Recrystallize from water, record the mp and submit your sample.
● Calculate the % yield and the molar proportions of reagents used.
● Outline the mechanism of the reaction and explain why conc. H2SO4 is used.
● Why was water added at the end of the reaction?

Experiment 18. Preparation of fumaric acid from maleic anhydride.

Method. Dissolve maleic anhydride (2.5 g) in boiling water (4 ml) in a test-tube to form a solution of
maleic acid. Cool the solution by placing the test-tube in a beaker of cold water. Filter the solid acid
using a Buchner funnel, but do not attempt to wash the solid. Dry it between pads of filter papers and
determine its melting point. Collect the filtrate, add concentrated HCl(6 ml) and reflux the mixture
for about 20 minutes. Crystals are formed which can be filtered, washed with water and dried
between pads of filterpaper. Determine the melting point. [mp of maleic acid: 130o; mp of fumaric
acid: 287o).
◦ Draw the configurations of maleic acid and fumaric acid.
◦ Provide their IUPAC names including their E,Z assignments.
◦ Provide a mechanism for the maleic anhydride- fumaric acid conversion.
 ◦ Which acid shows strong C=C absorption in IR?

Experiment 19. Preparation of Oximes.

Method. Dissolve KOH (1 g) in water (3.5 ml). Cool and add to a solution of hydroxylamine
hydrochloride (1.4 g) in water (3.5 ml) contained in a boiling tube. Place the mixture in an ice bath and
add Cyclohexanone (1 ml) in small quantities while shaking the mixture and keeping the temperature
about at about 10o. Allow to stand in the ice bath for 15 min, filter the crude acetoxime at the pump
and drain well. Record the mp and submit your product in a labelled test tube.
1. Record the mp of pure acetoxime.
2. Why did you add NaOH to the reaction mixture?
3. Calculate the molar proportions of reagents used.
4. Outline a mechanism for the reaction.

Experiment 20. Acetylation of an Amine or a Phenol.

Method. Add acetic anhydride (1.5 ml) to the amine/phenol (0.5 g) contained in a dry boiling tube. Add
2 drops of conc. H2SO4 mix well and warm in a hot water bath (50-60°) for 20 min. Allow to cool to
room temperature, then pour the mixture into a beaker containing about 20 ml of water and stir well.
Filter the solid at the pump, wash with cold water and drain well. Recrystallise with water.
Decolourise if necessary. Record the mp of your product and submit the sample. Consult mp tables
and try to identify the amine/phenol.
▪ Calculate the molar proportions of reagents used
▪ Outline a mechanism and indicate the role of conc. H2SO4 in the reaction.

Experiment 21. Benzoylation of an Amine or a Phenol

Method (a): Suspend/dissolve the amine/phenol (0.5 g) in of 10% NaOH (15 ml) solution contained in a
boiling tube. Add benzoyl chloride (1 ml), cork the tube and shake the mixture vigorously for 15 min.
Filter the crude product at the pump, break up any lumps on the filter with a spatula, wash thoroughly
with water and drain well. Recrystallise with methanol, decolourising if necessary. Record the mp.
Consult mp tables and try to identify the amine/phenol.
1. Outline a mechanism for the reaction you have carried out.
2. What are the functions of NaOH in the reaction?
3. Could you have prepared the acetate of an amine or phenol under the same conditions?
Explain.
Method(b): for amines (aniline) only Carry out the following steps in a fume hood. Measure equimolar
quantities of amine(aniline-1 ml) and benzoyl chloride(1.2 ml) (10 mmol) and mix them in a small
beaker with a glass rod . HCl gas will evolve and a solid mass will form. Add 10 g of crushed ice to the
contents of the beaker and stir well. The mixture will gradually soften and product will deposit on the
walls of the beaker. Stir until the product is completely precipitated and the supernatant is clear.
Filter the product, wash with water, dry well and recrystallize with a suitable solvent.
1. What are the functions of NaOH in the reaction?
2. Could you have prepared the acetate of an amine or phenol under the same conditions?
Explain.

Experiment 24. Oxidation of aromatic compounds.

a. Preparation of anthraquinone from anthracene

Place anthracene (0.5 g) in glacial acetic acid (5 ml) in a 50 ml round bottomed flask fitted with a
reflux water condenser and heat the flask so that the acetic acid boils gently. Dissolve chromium
trioxide(1 g) in water (1 ml) and add glacial acetic acid (2.5 ml) and pour this solution slowly down the
condenser. Shake the contents of the flasks and boil gently for 15 min. Allow to cool and then pour
the contents of the flask into an excess of cold water (10 ml). Crude anthraquinone separates as a
greenish-grey powder. Stir the mixture vigorously and then filter off the anthraquinone under gentle
suction. Wash thoroughly on the filter with hot water, and drain well. Purify anthraquinone by
sublimation (figure 7a). Submit your sample of pure anthraquinone. mp determination is not required.
1. What are the industrial applications of anthraquinone?
2. Why does the oxidation of anthracene give mainly anthra-9,10-quinone?

Experiment 26. Preparation of p-nitroaniline and its benzal (benzylidene) derivative

a. Acetylation of aniline (Expt. No. 20)

b. Nitration of acetanilide

Method. Dissolve acetanilide (2.5 g) in glacial acetic acid (3 ml). Add conc. H2SO4 (5 ml) and cool in an
ice bath. Prepare the nitration mixture by adding conc. H2SO4 (1 ml) to conc. HNO3 (1.2 ml) (CARE!) in
a test tube and cool in an ice bath. Stir the acetanilide solution and slowly add the nitrating mixture
through a dropper. During addition, the temperature of the reaction mixture should be kept below
10°. When all the nitrating acid has been added allow the reaction mixture to warm up to room
temperature and stand for a further 15 min. Then pour on to crushed ice (about 30 g). When the ice
has melted, filter off the crude p-nitroacetanilide, wash with cold water and dry by suction. Leave a
little crude product for TLC and recrystallise the remainder from methylated spirit. Spot the crude
product, pure crystals and the filtrate from recrystallisation separately on a TLC plate. Use CH2Cl2 as
the eluent and develop the plate. Compare the pattern of spots obtained from each sample. The
compounds are coloured and should be visible. The colours of the spots may be intensified by briefly
exposing the TLC plate to iodine vapours.
1. Record the mps of the crude and pure p-nitroacetanilide.
2. Calculate the % yield.
3. Sketch the TLC pattern observed. Account for the differences between the patterns of
spots observed on TLC for each sample.

Experiment 31. Carbanion additions

a. Preparation of sorbic acid from crotonaldehyde (Knoevenagel reaction)

Method: Place crotonaldehyde (2 g, 2.4 ml), dry malonic acid (3 g) and dry pyridine (3 g, 3.0 ml) in a 100
ml round bottomed flask, heat under reflux on a water bath, until the evolution of carbon dioxide
ceases (two hours). Cool the reaction product in ice, acidify with a solution of concentrated sulphuric
acid (1.1 ml) in water (2.5 ml). Cool in ice, filter, wash with water and recrystallise from water, to
obtain pure sorbic acid, mp 134°C.

b. Preparaton of dibenzalacetone (Claisen-Schmidt reaction)

Method. Prepare a mixture of benzaldehyde (1.5 ml) and acetone (0.5 ml). Add about half of this
mixture to a solution of NaOH (1 g) in ethanol (10 ml) and water (15 ml), contained in a conical flask.
Cork the flask and shake vigorously. Cool if necessary. A precipitate should appear in a few minutes.
After 15 minutes add the remaining half of the benzaldehyde-acetone mixture to the alkaline
solution. Shake for a further 30 min.. Filter the crude dibenzalacetone, wash with water and
recrystallise from ethanol-water to obtain pure dibenzalacetone, mp 112°C
 5. INDUSTRIAL APPLICATIONS

Dyes and Dyeing

Dyes are coloured because they absorb light in the visible region of the spectrum. Dyes
contain a chromophore, (unsaturated group like –N=N–, C=O, C=C) which absorbs in the UV region
of the spectrum and does not impart colour to a molecule. When the chromophore is part of a
conjugated system, absorption may occur in the visible region of the spectrum. Dyes may also contain
auxochromes (groups such as –NH2, –VR2, –OH and –OR). Auxochromes have at least one unshared
electron pair and increase the wavelength as well as the intensity of light absorption. Dyes may also
contain an anchoring group which helps to bind the dye to the substance being dyed by ionic, covalent
or hydrogen bonding. The –SO3H and –CO2H are grouped as common anchoring groups.
Dyes are often classified according to the different methods used for dyeing. Several methods
are needed because many different types of fibres are now in use. e.g., polyethylene of low polarity,
cotton with medium polarity, and wool which is a highly polar polypeptide with many free amino and
carboxyl groups. A dye should penetrate the fibre and remain firmly bound during washing and
cleaning.
Direct dyes remain fixed to the cloth without additional chemicals. Wool and silk which
contain many anionic polar sites, readily bind triphenylmethane dyes e.g., malachite green. Congo red
has polar amino and sulfonate groups which can hydrogen bond to hydroxyl groups and is a direct dye
used for cellulose fibres such as cotton and rayon.

Diazotisation and Preparation of Azo Dyes

Primary aromatic amines react with cold nitrous acid to form solutions of diazonium salts.
This process is known as diazotisation. Unlike the unstable aliphatic diazonium ions, aromatic
diazonium ions are stable at low temperatures. They can be coupled with phenols to give azo dyes.
The azo dyes are only slightly soluble in water.

Experiment 40. Preparation of Benzene-azo-2-naphthol.

Method: Dissolve aniline (0.5 ml) in a mixture of water (1 ml) and conc. HCl (1 ml) in a 50 ml beaker.
Cool to 0-5°C, stir well and add 10% NaNO2 solution (3 ml) in small portions to form the diazonium
chloride. Maintain the temperature below 5°C during this addition.
Dissolve 2-naphthol (0.5 g) in 10% NaOH (2.5 ml) and cool to 0-5 °C. Add the diazonium
chloride solution slowly and with stirring to the cooled naphthol solution. Stir and allow the reaction
mixture to remain at 0-5 °C for a further 20 minutes and then filter the red crystalline product under
gentle suction. Wash with water, press the residue on to the filter with a spatula and drain well.
Submit the crude, dry product. Recrystallise a small amount from ethanol and record the mp of the
pure, dry crystals.
1.Calculate the molar proportions of the reagents used (2-naphthol, aniline, HCl & NaNO2). Explain
why these proportions are used.
2.Give the structure of benzene-azo-2-naphthol and explain why it is the product of the reaction.