EXPERIMENT 2.

SYNTHESIS OF AN ESTER BY FISCHER

ESTERIFICATION: SYNTHESIS OF ISOAMYL ACETATE

INTRODUCTION AND THEORETICAL BACKGROUND

Esters are a group of compounds of general formula R-CO-O-R', which are widely
distributed in nature. Simple esters usually have a pleasant smell. In many cases, the
characteristic aroma and taste of fruits and some flowers are due to compounds that have
an ester group in the structure. The organoleptic properties of many aromas are due to a
predominant single ester. This is the case of essential banana oil, of which isoamyl acetate
is the main constituent. In most cases, however, the aroma is due to a complex mixture of
esters and other compounds such as terpenes.

The triglyceride ester group is the main functional group of lipids and their derivatives (fats,
oils, waxes, etc.).It is also widely represented among substances in therapeutic applications
such as local anaesthetics (structurally derived from the natural alkaloid cocaine), some food
and cosmetic preservatives such as p-hydroxybenzoic acid derivatives and many others.

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Esters are usually prepared by reaction between a carboxylic acid derivative and an alcohol.
The reaction of an alcohol with an acid chloride or an acid anhydride is produced rapidly and
quantitatively due to the high electrophilicity of the carbonyl group of these acyl derivatives.
In these cases, the reaction proceeds in almost stoichiometric conditions.

Although this method is widely used in the laboratory, industry prefers the reaction between
alcohols and carboxylic acids, which are much cheaper.
The reaction of an alcohol with a carboxylic acid is known as Fischer esterification and is a
very slow equilibrium reaction. By adding a catalytic amount of a mineral acid such as
sulphuric, hydrochloric or phosphoric acid, both components react quickly to give an ester
and water. This is not a very exothermic transformation and the formation of the ester
proceeds to reach a maximum equilibrium concentration which is ruled by the
thermodynamic equilibrium constant (Keq)

In the mixture, we will find all components – carboxylic acid, alcohol, ester and water – and
therefore we must shift the equilibrium to the right if it is to be synthetically useful.
To do this, we can use a large excess of one of the reagents (acid or alcohol) or eliminate
the water as it is formed. Fischer's esterification is also a good laboratory synthetic method
when carboxylic acid (acetic acid, propionic) and the alcohol to be used (methanol, ethanol,
butanol, etc.) are cheap and available in large quantities (an excess can be used with no
problem).

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In this experiment we will prepare isoamyl acetate by direct esterification (Fischer
esterification) of the isoamyl alcohol (3-methylbutan-1-ol) with acetic acid. As well as being
the main component of the banana aroma, isoamyl acetate is also the alarm pheromone of
bees.
Fischer's esterification follows the general mechanism below:
Initially, the protonation of carbonyl oxygen is produced by generating more reactive species
stabilized by resonance.

Carbonyl carbon in this species is susceptible to being attacked by the hydroxyl group of the
alcohol to produce a tetrahedral intermediate. This intermediate evolves towards the ester
formation. On the contrary sense, protonation of the oxygen in the isoamyl group induces
loosing of isoamyl alcohol. In the other cases, the protonation of either of the two hydroxylic
oxygens induces the removal of water and the formation of the ester. The mechanism can be
summarized in the following stages:

This mechanism is the inverse of that found in the acid hydrolysis of esters when heating an
ester with an excess of water and acid medium. In this case, we shift the equilibrium to the
left. The hydrolysis of esters, which can also be catalysed by bases, is also of great synthetic
utility and serves, for example, to obtain fatty acids from triglycerides.

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TIMING OF THE EXPERIMENT: 2 sessions
REAGENTS TO BE USED:
Isoamyl alcohol (15.9 g) FLAMMABLE, TOXIC
Glacial acetic acid (23 mL) FLAMMABLE, IRRITANT
Concentrated sulphuric acid (4 mL) CORROSIVE (CAUSES SEVERE BURNS)

LIST OF TECHNIQUES AND BASIC PROCEDURES:

- Reaction with a reflux condenser

- Shifting an equilibrium reaction with an excess of reagent
- Using a catalyst
- Extraction of a mixture by chemical reaction
- Drying an organic phase with a drying agent
- Simple distillation at atmospheric pressure
- Boiling point

EXPERIMENTAL PROCEDURE

In a 100 mL round-bottomed flask, place 15.9 g (0.18 mol) isoamyl alcohol and then add 25
mL (0.40 mol) of glacial acetic acid (note 1). Whilst constantly stirring this mixture, add 4 mL
of concentrated sulphuric acid (note 2) dropwise.
After adding some pumice stones, heat the mixture at reflux for 1 hour, and then cool to
room temperature.
Transfer the crude reaction mixture to a separatory funnel (note 3) and add 50 mL of cold
water. Wash the flask with 10 mL of cold water and add it into the funnel. Cover the
separatory funnel and shake it several times. Separate the aqueous layer from the organic
layer (notes 4 and 5). Wash the organic layer with a 5% NaHCO3 solution (approximately 30
mL). To do this, first gently stir both layers with the uncovered funnel until no more carbon
dioxide detachment is observed (note 6). Cover the funnel and shake it a couple of times,
leaving the gas out. Continue shaking until there is no CO2 detachment. Remove the
aqueous layer (note 5) and repeat the process by adding a new portion of sodium
bicarbonate solution until the aqueous phase has an alkaline pH. Finally, wash the organic
phase with 25 mL of water, and add some NaCl if the two-layer separation is not good
enough (note 7).
Separate and dry the organic layer using anhydrous magnesium sulphate (note 8).
Filter the organic solution through filtering paper using a conical stemmed funnel (note 9).
The filtrate has to be collected directly in a dry distillation flask of appropriate size. Next, add
some porous stone and fit a dry distillation apparatus (note 10). Upon gradual heating,
collect the fraction distilling between 134°C and 141°C (note 11) in a previously weighed
flask in an ice-water bath to prevent evaporation losses.

Weigh the product and calculate the yield.

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NOTES:

Note 1: Equilibrium can also be shifted towards the formation of ester by using an excess of
isoamyl alcohol. In this case, an excess of acetic acid is used not only is it cheaper but it also
greatly simplifies the process of subsequent separation and purification because of its
solubility in water.
Note 2: Sulphuric acid is a very strong acid that releases a considerable amount of heat
when it is diluted in water or other solvents. If there is too little solvent, the heat produces
sudden evaporation and acid splints. For this reason, you should never add water or other
products to sulphuric acid but vice versa. Sulphuric acid also promotes dehydration and
carbonizing organic matter when highly concentrated. This may cause the reaction mixture to
darken and the formation of secondary products that are not always easy to eliminate. This
is worsened by the fact that sulphuric acid is very dense and tends to accumulate and form a
bottom layer in the flask. For this reason, sulphuric acid should always be added dropwise
gently stirring the content of the flask to ensure that at any moment the mixture is completely
homogeneous. If a large amount of acid has to be added or if the solution is self heated on
mixing, it may be necessary to cool the flask externally using a water bath.
To overcome these problems in organic synthesis, other less reactive but also strong acid
catalysts are commonly used, such as H3PO4 (orthophosphoric acid) or sulfonic acids such
as CH3C6H4SO3H (p-toluenesulphonic acid).
Note 3: Extraction is the most used technique for separating a specific organic compound
from a reaction mixture or for isolating it from natural sources. In general, it can be defined
as the separation of a component with the help of a solvent.
In practice, extraction with a solvent is widely used to separate (to extract) organic products
from the suspensions or watery solutions in which they are found. The technique of
extraction is also used to remove excess reagents, catalysts or by-products from the reaction
mixture, providing that they are more soluble in water than in organic solvents. This
procedure is called "washing". Liquid-liquid extraction is usually done with the help of an
extraction funnel (separatory funnel) and it basically consists of introducing the solvent (or
organic phase) and water (or the appropriate aqueous solution) and shaking. When finished,
the funnel is supported on a metal O-ring, allow the two layers to separate and remove the
lower layer by carefully opening the bottom key.

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In general, when a mixture of organic compounds is combined with a mixture of two
immiscible solvents (one of which is usually aqueous) the different compounds are
distributed between the two phases according to their relative solubility.
The ratio of concentrations between the two phases C org and Caq is known as the distribution
coefficient Kp = Corg / Caq. Depending on the nature and relative solubility of the compounds,
this coefficient may or may not be very large. If it is, the whole product will be found in the
organic phase and it will be separated with only one extraction. If it is not, much more
common, the extraction will have to be repeated with new portions of solvent to ensure that
the product has been fully recovered. If this parameter is not known, it is always better to
make several extractions to ensure that the whole product has been recovered.
When washing reaction mixtures it is very common to extract reagents or catalysts by means
of a chemical reaction. If the product that we want to remove is an acid or a base, we will do
the washing with a basic or acid solution, respectively. The acid-base neutralization will give
a salt that is extremely soluble in water and, therefore, the Kp will be very favourable.
Some important aspects that need to be taken into account in the extraction operations are:
a) The extraction funnel: It must be of the appropriate size and that the key and the cap
must adjust perfectly to ensure no losses. It must be clean and very well rinsed; traces of
soap favour the formation of emulsions. Normally, since one of the phases is usually
aqueous, it does not need to be dried. However, if it is not, it must also be well dried (for
instance, extractions with concentrated sulphuric acid in experiment 3).
b) The extraction funnel must always be kept with the key locked and the lid open in a
vertical position using a support and an O-ring. The stopper is only fitted to the funnel
when we are shaking it to perform the extraction. After use, the funnel goes back to the O-
ring and the stopper is removed.
c) The extraction operation is done as illustrated below:

d) The funnel must be shaken very gently at the beginning so that it can generate gases
or solvent overpressure. These overpressures must be released by inverting the funnel as
shown in the figure above and opening the key.

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 Overpressures are very common in two cases: 1) Extraction with low-boiling point
solvents: diethyl ether (37ºC) and pentane (40ºC). Never extract a warm solution with
these solvents. 2) Extraction with carbonate or bicarbonate solutions to neutralize acids
(as in this experiment), since it produces a large amount of CO2. In this case, the funnel
should gently swing in a vertical position with the stopper open. The funnel should only be
covered and stirred when the reaction is over and there is no strong CO2 evolving.

Note 4: In most cases the two layers separate in a few minutes to form a fine interface that
them to be distinguished and the separation process to begin (a) In other cases, emulsions
are formed that are difficult or impossible to get rid of (especially if there are surfactants such
as soap) (b) The formation of emulsions depends on the structure of the compounds, the
nature of the solvent, the concentration of the different species and sometimes the pH of the
medium. Emulsions are usually a single layer that is milky in colour or two clear layers with a
thick milky interface in between. This prevents the layers from being separated.

a) b)

When an emulsion is formed it must be separated into two phases with a clear interface.
There are several techniques for doing what is called "breaking the emulsion":
a) Modify the ionic strength of the aqueous phase: the emulsions are droplets of two
phases dispersed in each other, which remain stable and separate thanks to surface
charges that make the droplets repel each other. Adding an ionic salt (usually sodium
chloride) to the aqueous phase increases its ionic strength and breaks the equilibrium.
To do this, add some salt (ensure there is no salt in the funnel neck), put the stopper in,
and shake until the salt dissolves. Return the funnel to the vertical position, remove the
stopper and see if the emulsion has broken.

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 b) Change solvent density: in the emulsion, the droplets stay in suspension because they
have a similar density to that of the other phase. Adding a particular amount of a dense
solvent (for example, chloroform) or a low density solvent (for example, diethyl ether) can
break the equilibrium and separate the emulsion
c) Vary surface tension: a change in surface tension can favour the breaking of the
droplets and cause them to come together. Normally, some ethyl ether (which has a very
low surface tension) is added.
When choosing the extraction solvent, it should be taken into account that some solvents
have a greater tendency to form emulsions than others. Ethyl ether hardly forms emulsions
at all so, together with the ease with which it evaporates, it is one of the most widely used
extraction solvents. Chloroform, dichloromethane and sometimes toluene are more likely to
form emulsions mainly in basic media.

Note 5: Once the layers have been separated, you must know where the product is, and
which layers are the organic and the aqueous one. Normally this can be determined
empirically if you know the solvent density used. For this reason, bear in mind that all the
usual organic solvents (ethyl ether, ethyl acetate, toluene, hexane, etc.) are less dense than
water with the exeption of halogenated ones (chloroform, dichloromethane, carbon
tetrachloride, dichloroethane etc). Care must be taken that in aqueous solutions with a very
high concentration of salts (NaCl, NaOH, Na2CO3, etc.), the density of the aqueous phase
can be significantly greater than 1.0 and invert the order of the two phases. If in doubt, add a
drop of water and observe if it stays in the upper layer or goes through it and dissolves in the
lower part. In addition, add a few millilitres of water with a Pasteur pipette and observe which
layer increases in volume. This test is conclusive.
note 6: See section d) in note 3.
Note 7: Whenever acid or basic aqueous solutions are used for the final washing, one or two
washings with water must subsequently be done to remove the remains of these reagents. If
the organic product is slightly soluble in water, washing is done with a saturated NaCl
solution (brine), so the solubility of the compound is significantly reduced and losses are
minimized.
Note 8: The organic compounds and their solutions resulting from an extraction process are
saturated with moisture and must be dried carefully before isolation. To do this, add a small
amount (5-10% in total volume) of a non-reactive anhydrous salt. Stir the mixture for a few
minutes and observe the formation of lumps. We know that we have used the appropriate
amount of drying agent when it remains in powder form after addition. There are several
neutral drying agents, usually dehydrated inorganic salts. Some of the most widely used are
anhydrous sodium sulphate and anhydrous magnesium sulphate. In contact with water,
these salts quickly incorporate them into their structure as hydration waters. In particular,
anhydrous magnesium sulphate can react with 8 molecules of water to form the MgSO4 •
8H2O. There are many other drying agents and each solvent has its own drying agents. You
need to become familiar with the main ones.

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Note 9: To separate magnesium sulphate or the drying agent used, filtration by gravity is
commonly used through regular filter paper (not necessarily folded). To avoid losses in the
organic phase, once the filtration has finished, you can carefully press the filter paper to
drain what is absorbed in the drying agent. If you are filtering the solution of an organic
compound in a solvent, you can also wash the drying agent and the filter paper with a small
amount of the same solvent.
Note 10: Distillation is the most common method for purifying liquids. Depending on the
boiling point of liquids and their composition, different types of distillations can be done.
Simple distillation is used for products with a single volatile component, and fractional
distillation is used to separate volatile compounds that have different boiling points (a
difference of at least 20-30ºC). If the compound’s boiling point is less than 200-250°C, the
distillation can be made at atmospheric pressure but if it is higher, it should be done at
reduced pressure distillation (vacuum distillation) so that it does not decompose. Simple
distillation is always used to separate a volatile liquid (usually the reaction product) from the
non-volatile impurities.
The basic equipment used consists of at least six pieces: a round flask, a Claisen head (or a
three-armed piece with an elbow at 75°) with a thermometer connected to the top (either
directly or through a screw adapter or Quickfit ).

A Liebig-West or tubular condenser is connected to the inclined arm. (Remember that the
water enters the condenser at the lowest point). Finally, a receiving collector is placed to
collect the distillate. It is advisable to place an elevator under the heating bath so that the
distillation speed can be regulated. The system should always start to be assembled from
the flask, on the plate or heating bath above the elevator, as it is this that determines the
position of the whole system. Before assembling the rest of the system, the flask must be
filled with the product to be distilled, the porous stone or, alternatively, a stirring rod. The
Claisen piece is then connected to the thermometer and the condenser, in this order. At least
two clamps and two vertical stands are required to hold the system, one by the flask neck
and the other, inclined, at the midpoint of the condenser. Finally, a receiver elbow is attached
with a Keck clip or a clamping thread (if the collector has a glass screw).

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The distillation flask must be heated progressively. The distillate that boils within the correct
boiling range must be collected. If the product to be distilled is not completely dry, at first a
milky liquid forms which is a mixture of water and the product (or solvent remains). This
fraction should always be rejected and only the clear liquid should be collected.
Note 11: The position of the mercury bulb in the thermometer is very important for correctly
reading the correct distillation temperature. If the volume of liquid to be distilled is relatively
small (5-25 mL), the upper part of the mercury bulb must be more or less at the same height
as the lower point of the elbow output. This reduces the amount of distillate not collected
while the thermometer is equilibrating the temperature with the incoming vapour.

It should be taken into account that the thermometer will not mark the actual temperature of
the liquid until vapours does not completely wet the lower part of the thermometer and
reaches thermal equilibrium (which may take one or two minutes). It should also be taken
into account that at the end of the distillation, the vapour is overheated and the temperature
may be slightly above the boiling point of the pure liquid.
When larger liquid volumes are distilled, the volume of vapour is greater and the
thermometer is usually placed in the middle of the outlet tube. In this case, the volume of
distillate lost is not significant.
The thermometers commonly used in distillation equipment usually have zero displacement
(the scale starts at a point above zero) and there are two types: rod thermometer and Opal
scale thermometer. Probes and electronic thermometers are not commonly used because of
the corrosive effects of the hot vapours.

Rod thermometer Opal scale thermometer

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QUESTIONS AND TASKS

1. What is the role of the sulphuric acid added to the reaction mixture? Why is the acetic acid
itself not as effective as a catalyst?
2. What is the limiting reactant in the reaction? Would it be appropriate to invert the
proportions of the reagents? Find the solubility in water and the boiling point of all the
components to explain your answer.
3. Why do we wash the organic solution with a solution of NaHCO3?
4. How does anhydrous magnesium sulphate work in the drying of liquids and solutions?
5. Make a list of at least 6 different common solvents and their appropriate drying agents.
6. Write the mechanism of the acid hydrolysis of an ester.
7. Propose two alternative methods for synthesizing an ester, other than nucleophilic acyl
substitution.

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