Irene Martínez Pérez

ISOAMYL ACETATE SYNTHESIS

TREBALL DE FI DE GRAU

dirigit pel Sr. Herman Faes

Grau de Química

Lovaina

2018
ACKNOWLEDGEMENTS
This study was carried out in the facilities of the "University College Leuven-Limburg"
(UCLL), during an Erasmus mobility program, with the supervision of Herman Faes,
organic chemist, also member of the group “Technology Department of Sustainable
Chemical Processes” at KUL.

First of all, I want to thank my supervisor, professor Herman Faes, for accepting me on
this Erasmus semester and helping me in everything I have need, and also for teaching
me and allowing me to learn a lot about Organic Chemistry, as well as Belgium. His
energy and sense of work are an example to be followed.

I had a great support and advice throughout this study and I would like to acknowledge
to the UCLL for their contributions.

Furthermore, I would like to acknowledge to the “Universitat Rovira i Virgili” for

offering me the opportunity to do an Erasmus mobility program and also to my
mobility coordinator, Elena Fernández, for helping me from a distance in my
adaptation to the new university and environment.

And last, gratefulness to my family and friends who, even thousands of kilometres
away, they have always supported me.

For all of you, just say it has been an honour staying at Belgium with you in my final
path of the chemistry bachelor.
TABLE OF CONTENTS

1. SUMMARY................................................................................................................. 1
2. LIST OF ABBREVIATIONS AND SYMBOLS ................................................................. 3
3. OBJECTIVES ............................................................................................................... 4
4. INTRODUCTION ........................................................................................................ 6
5. LITERATURE REVIEW ............................................................................................... 13
6. MATERIALS AND METHODOLOGY .......................................................................... 21
6.1. CHEMICAL PRODUCTS...................................................................................... 21
6.2. MATERIAL ......................................................................................................... 25
6.3. CHARACTERIZATION EQUIPMENT ................................................................... 26
6.4. METHODOLOGY ............................................................................................... 26
6.4.1. Sulfuric Acid Catalyst ................................................................................ 26
6.4.2. p-Toluenesulfonic Acid Catalyst ............................................................... 27
6.4.3. Amberlyst-15 Catalyst .............................................................................. 28
6.4.4. Molecular sieves ....................................................................................... 28
6.4.5. Azeotropic distillation ............................................................................... 29
6.4.6. Steglich reaction ....................................................................................... 30
7. RESULTS AND DISCUSSION ..................................................................................... 32
7.1. CATALYST ......................................................................................................... 32
7.2. REMOVAL OF WATER ....................................................................................... 34
7.3. STEGLICH ESTERIFICATION ............................................................................... 36
7.4. ISOAMYL ACETATE CHARACTERIZATION ......................................................... 37
8. CONCLUSIONS......................................................................................................... 39
9. LIST OF REFERENCES ............................................................................................... 40
APPENDIX A .................................................................................................................... 42
APPENDIX B..................................................................................................................... 43
ISOAMYL ACETATE ANALYTICAL STANDARD 1H NMR SPECTRA ................................. 43
ISOAMYL ACETATE 1H NMR SPECTRA ......................................................................... 44
APPENDIX C..................................................................................................................... 45
ISOAMYL ACETATE ANALYTICAL STANDARD INFRARED SPECTRA .............................. 45
ISOAMYL ACETATE INFRARED SPECTRA ..................................................................... 46
 1. SUMMARY
The synthesis of esters has played a most important role in organic synthesis from its
start. Esters constitute functional groups of chemical significance, in numerous natural
and synthetic compounds, and also they have utility in diverse fields both in the
industry and in laboratory. The essential feature of esterification, that particularly
distinguishes it from other reactions, lies in its broad utilization in industry. Just a brief
look reminds us of aspirin (acetyl salicylic acid), fatty acid esters, polyesters...

A variety of routes to arrive at esters are possible, and numerous methods have been
reported. Since esterification has such a long history and the reaction itself is simple,
many people take it for granted that little room is left for further scientific
improvements. On the other hand, in industry, esterification still has permanent
significance and so many new technologies remain undisclosed. Esterification has in
fact been, and is still undergoing, extensive innovations.

The biggest problem frequently encountered in esterification technology arises from

equilibration. The condensation between an alcohol and an acid is slow and reversible,
thus the equilibrium can be shifted to the desired product by adding excess of one of
the reagents or by the removal of products.

On the other hand, the rate of the reaction can be influenced by adding a catalyst such
as an acid. Mineral liquid acids, Lewis acids and Bronsted acids are widely used in
industry. However, their corrosive and hazardous nature, the subsequent need for
neutralization generating large quantities of aqueous waste, and the difficulty to
separate them from the reaction medium restrict their industrial applicability.

Innovations to further improve the efficiency and sustainability of the esterification

process are highly desirable. To accomplish a green esterification protocol, recoverable
heterogeneous solid catalysts are used.

Solid acid catalysts have received much attention as potential replacements for
mineral liquid acids. The advantages of heterogeneous solid acid catalysts include
reduction of corrosion, ease of separation from the reaction medium, good
recyclability and enhanced product selectivity.

In addition to being essential molecular components in their own right, ester groups
also play versatile temporary roles in organic synthesis for protection of carboxylic
acids and hydroxyl groups. The synthesis of natural products, especially macrolides,
sugars, and peptides, depends heavily on acylation technology.

1
Furthermore, the low-molecular-weight esters have very pleasant odours and indeed
are the major components of the flavour and odour aspects of a number of fruits.
Although the natural flavour may contain nearly a hundred different compounds,
single esters approximate the natural odours and are often used in the food industry
for artificial flavours and fragrances.

2
 2. LIST OF ABBREVIATIONS AND SYMBOLS

TsOH p-Toluenesulfonic acid

DCC Dicyclohexylcarbodiimide

DMAP 4-dimethylaminopyridine

DHU Diclyclohexylurea

DCM Dichloromethane

3
 3. OBJECTIVES

The ester group is an important functional group that can be synthesized in a number
of different ways. Fischer-Speier esterification [1] is a good method of synthesis in the
laboratory when carboxylic acid or alcohol to be used is an economic product and
available in large quantities.

The aim of this project is the synthesis of a specific ester, the isoamyl acetate, which is
obtained by the condensation of isoamyl acohol with acetic acid through a Fischer-
Speier esterification. This ester is the main constituent of the essential banana oil.

The synthesis of esters directly from alcohols and carboxylic acids plays an important
role in organic synthesis because of its utility in diverse fields both in academia and in
industry. For this reason, the main objective of this final thesis is to develop new
methods to obtain the maximum conversion of the isoamyl acetate synthesis. In this
way, the best reaction conditions for this shynthesis will be known. To achieve this
goal, different pathways will be followed.

1) Different acid catalysts can be used to increase the rate of the reaction. In this
study are used sulfuric acid, p-Toluenesulfonic acid (TsOH) and Amberlyst-15.
The last one is a solid acid catalyst, which has received much attention as
potential replacement for mineral liquid acids, due to its mild and highly
selective properties combined with its environmentally benign character.

2) The use of the Chatelier’s Principle to shift the equilibrium towards products,
by increasing the concentration of the reactants.

3) The use of the Chatelier’s Principle to shift the equilibrium towards products,
by decreasing the concentration of a product by removing it from the
equilibrium. In this case, the water will be absorbed and removed from the
reaction mixture through two techniques.

a. By adding to the reaction mixture molecular sieves, an artificial zeolite,

which preferentially adsorbs water. Two types of molecular sieves (3A
and 4A) are used either in the reaction mixture and also placed above
the reaction mixture.

b. By an azeotropic distillation with toluene. Water can form an azeotrope

with certain alcohols at specific concentrations. A special apparatus,
called a Dean Stark apparatus or trap, can be used to distil off the
water/toluene azeotrope while the ester being is formed. Removing
water from the reaction in this way will favour ester formation.

4
4) The use of carbodiimide activators through the Steglich Esterification [2]. It is
an esterification variation with dicyclohexylcarbodiimide (DCC) as a coupling
reagent and 4-N,N-dimethyilaminopyridine (DMAP) as a catalyst. The Steglich
Esterification has been used to facilitate the esterification of acids and alcohols
and does not require high temperatures.

5
 4. INTRODUCTION

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,
although not all, the characteristic aroma and taste of fruits and some flowers are due
to compounds that have the structure of the ester group (Table 1). The organoleptic
properties of many aromas are due to a single ester which predominates as is the case
with essential banana oil in which the main constituent is isoamyl acetate. In most
cases however, the aroma is due to a complex mixture of esters and other compounds
such as terpenes, norisoprenoids (e.g. damascenone), volatile phenols and other
benzene derivatives (raspberry ketone), monoterpenes (e.g. linalool, nerol, geraniol...)
Table 1. Fragances and boiling point of esters [3]

Boiling point
Ester and formula Fragance
(ºC)

101.7
Pear

98.4 Raspberry

102.3 Apple

121 Pineapple

136.8 Rum

6
 142 Banana

206 Peach

210 Orange

222 Wintergreen

For instance, esters are the class of volatile compounds that are responsible for a fruity
smell in wines. Wine is commonly referred to as a “complex matrix” and this is
certainly true. However, by breaking wine down into its fundamental components, it is
possible to understand how to better manage the vineyards and wineries to attain the
wine styles that the market desire. Esters could be considered as a whole of aromas
that are very valuable to the wine industry, but only the aliphatic monocarboxylic
esters make a significant impact in it [4].

Most of the esters in wine are formed during fermentation or during wine ageing. In
particular, esters are critical in the production of Pinotage wine. If uncontrolled, this
variety develops a pungent banana aroma from isoamyl acetate, which is not only
produced during fermentation, but also found within the grape itself. This ester is
formed from acetic acid and alcohol. The mechanism by which yeasts form esters is
believed to be a reaction catalysed by an enzyme called alcohol acetyltransferase
(AAT). This reaction uses alcohol as a substrate, co-enzyme A and ATP to form an ester.
Esters may also be formed through simple hydrogen-ion-catalysed reactions [5].

7
Furthermore, the triglyceride ester group is the main functional group of lipids and
their derivatives (fats, oils, waxes...). It is also widely represented among substances
with therapeutic use such as local anaesthetics derived from structural cocaine natural
alkaloids (Table 2) and in some food and cosmetics preservatives.
Table 2. Chemical structures of cocaine and some local anaesthetics derived from cocaine.

Cocaine

Lidocaine

Bupivacaine

Esters are used as protecting groups in synthesis to obtain chemoselectivity in a

subsequent chemical reaction. The protecting group is introduced into a molecule by
chemical modification of a functional group, to temporally mask the characteristic
chemistry of the functional group because it interferes with another reaction. The
most reactive functional groups commonly requiring protection are alcohols,
aldehydes, ketones, carboxylic acids and amines. Esters are the common protecting
groups for carboxylic acids, which have an acidic proton that can be abstracted by
bases including organometallic reagents.

8
A good protecting group should be readily, but selectively introduced to the desired
functional group in a poly-functional molecule. It should be stable to the reagents
employed in subsequent reactions steps in which the group being masked is desired to
remain protected. Moreover, it should be capable of being selectively removed under
mild conditions when its protection is no longer required.

For instance, in peptide synthesis due to the possibility of unintended reactions,

protecting groups are usually necessary. Peptide synthesis is the production of
peptides, which are organic compounds in which multiple amino acids are linked via
amide bonds, also known as peptide bonds. Peptides can be synthesized by coupling
the carboxyl group of one amino acid to the amino group of another amino acid
molecule (Scheme 1).

The use of protecting groups is essential during peptide synthesis to avoid side
reactions such as self-coupling of the activated amino acid leading to polymerization.
This would compete with the intended peptide coupling reaction, resulting in low yield
or even complete failure to synthesize the desired peptide. When protecting groups
are used, the unprotected amine of one reacts with the unprotected carboxylic acid
group of the other to form a peptide bond.

Scheme 1. Formation of a peptide bond using protecting groups

Esters are usually prepared by reaction between an acid derivative and an alcohol. An
organic base such as pyridine is usually added to react with the hydrogen chloride
formed. The reaction of an alcohol with an acid chloride or acid anhydride is produced
quickly and quantitatively due to the large electrophilicity of the carbonyl group of
these compounds so that it can work almost stoichiometric conditions (Scheme 2).

Scheme 2. Conversion of acid derivative to ester.

9
Although this is a widely used method in the laboratory, in the industry the reaction of
alcohols with carboxylic acids is preferred. Acid chlorides form hydrogen chloride upon
contact with atmospheric moisture, are corrosive and react often dangerously with
nucleophiles.

The reaction of an alcohol with a carboxylic acid is known by the name of Fischer-
Speier esterification and is a reaction of equilibrium that occurs very slowly because of
the little electrophilicity of the carboxyl group (Scheme 3). If a catalytic amount of a
mineral acid such as sulfuric, hydrochloric or phosphoric acid is added, the two
components react faster to form an ester and water. This transformation is not very
exothermic and the ester forms up to a balance concentration.

Scheme 3. Conversion of carboxylic acid to ester.

This transformation is exothermic or endothermic depending on the esters that are

formed. Hess’s law can be used to determine the overall energy required or released
for a chemical reaction. Allows the enthalpy change for a reaction to be calculated
even when it cannot be measured directly, using previously determined values for the
enthalpies of formation (Table 3). The esterification of isoamyl alcohol and acetic acid
to form isoamyl acetate will be used as an example.
Table 3. Standard enthalpies of formation of the compounds in isoamyl acetate synthesis [6].

Compound Δf H0 (kJ/mol)
Isoamyl alcohol -356
Acetic acid -484
Isoamyl acetate -438
Water -286

For Hess’s Law

𝛥𝑟 𝐻 0 = ∑ 𝛥𝑓 𝐻 0 − ∑ 𝛥𝑓 𝐻 0 =
𝑝𝑟𝑜𝑑𝑢𝑐𝑡𝑠 𝑟𝑒𝑎𝑐𝑡𝑎𝑛𝑡𝑠

𝑘𝐽 𝑘𝐽 𝑘𝐽 𝑘𝐽
= (−438 + (−286 )) − (−356 + (−484 )) =
𝑚𝑜𝑙 𝑚𝑜𝑙 𝑚𝑜𝑙 𝑚𝑜𝑙
𝑘𝐽
= 116
𝑚𝑜𝑙
As the enthalpy change is positive, this reaction is endothermic.

10
Fischer-Speier esterification is also a good method of synthesis in the laboratory when
carboxylic acid or alcohol to be used is an economic product and available in large
quantities.

Different acid catalysts can be used to increase the rate of the reaction. In this project
sulfuric acid, p-Toluenesulfonic acid (TsOH) and Amberlyst-15 are used. In the first
place, sulfuric acid is a strong mineral acid (pKa of -3.0) widely used in chemical
industry.

In the second place, TsOH is an organic compound soluble in water, alcohols and polar
organic solvents. It is a strong organic acid (pKa of -2.8) and unlike some strong mineral
acids as sulfuric acid, TsOH is non-oxidizing. These two catalysts are widely used in
industry, however their corrosive and hazardous nature, the subsequent need for
neutralization generating large quantities of aqueous waste, and the difficulty to
separate them from the reaction medium restrict their industrial applicability.

Solid acid catalysts have received much attention as potential replacements for
mineral liquid acids. The advantages of heterogeneous solid acid catalysts include ease
of separation from the reaction medium, good recyclability, reduction of corrosion and
enhanced product selectivity. The isolation and reuse of this catalyst is an example of
“Green Chemistry”, reducing consumption of non-renewable resources.

An example of heterogeneous solid acid catalyst is Amberlyst-15, a powerful and

selective acid catalyst for esterification reactions. It has played an important role in
organic synthesis, with versatile synthetic applications in different chemical
transformations as esterification. The growing interest in this catalyst is mainly due to
its mild and highly selective properties combined with its environmentally benign
character and commercial availability.

Additionally, this catalyst can be regenerated and recycled and is used in organic
synthesis as heterogeneous reusable acid catalyst. The maximum operating
temperature is 120ºC and the capacity is 4.7 mmol H+/g [7].

Amberlyst-15 is a macro reticular polystyrene based ion exchange resin with strongly
acidic sulfonic group (Table 4). It serves as an excellent source of strong acid and is
used in acid catalysed reactions as esterification. It is easy to measure, safe to use and
readily removed at the end of the reaction. [8].

11
Table 4. Different catalysts structures.

Sulfuric Acid p-Toluenesulfonic Acid Amberlyst-15

12
 5. LITERATURE REVIEW

The aim of this project is the synthesis of the isoamyl acetate by direct Fischer-Speier
esterification of acetic acid and commercial isoamyl alcohol (Scheme 4). The isoamyl
acetate, in addition to being the main component of the banana aroma, it also
constitutes the bee alarm pheromone, the release of which is a defensive reaction to
alert nearby bees. Its uses are as fruits (banana, pear, apple, berry) flavour for foods,
candy, butter, coconut, cola and rum; and also flavour for paint and insecticide.

Scheme 4. Synthesis reaction of isopentyl acetate.

Fischer-Speier esterification proceeds according to the following general mechanism

(Scheme 5). Initially, the protonation of carboxyl oxygen is produced by generating a
more reactive species that is stabilized by resonance. The carboxyl carbon develops a
full positive charge that is attacked by the nucleophilic alcohol group, giving a
tetrahedral intermediate. This tetrahedral intermediate with shorter bonds can either
lead to the formation of the ester or go backwards to the starting products. The
protonation of either of the two hydroxyls causes the removal of water followed by
loss of the catalysing proton and the formation of the ester.

13
Scheme 5. Reaction mechanism of the synthesis of isoamyl acetate

Because each of these steps is completely reversible, this process is also, in reverse,
the mechanism for the hydrolysis of an ester. The acid hydrolysis reaction of esters
takes place when heating an ester with excess water and acid medium. In this case, the
equilibrium is shifted to the left. This reaction of hydrolysis of esters, which can also be
catalysed by bases, also has a great synthetic utility and serves, for example, to obtain
the fatty acids from the triglycerides.

The equilibrium expression for this reaction is the following:

𝐾𝑒𝑞 =

For primary alcohols reacting with unhindered carboxylic acids, the K eq is

approximately of 4%. If equal quantities of the alcohol and acetic acid are allowed to
react, the theoretical yield of ester is only 67% at equilibrium. [3]

14
As a matter of fact, the Fischer-Speier esterification reaction takes advantage of Le
Chatelier’s principle to increase the amount of alcohol that is esterified. Chatelier’s
Principle is when one side of an equilibrium is altered by changing concentration or
pressure and the equilibrium shifts to counteract this change and restore the
equilibrium.

The equilibrium is shifted towards products by two different techniques:

1) Using a large excess of one of the reactants. The equilibrium shifts to use up
the increased amount of reactant, thereby producing more products.
2) Decreasing the concentration of a product by removing it from the equilibrium.
This causes more reactants to be consumed to restore the product
concentration.

In the case of isoamyl acetate synthesis, is used an excess of acetic acid because it is
cheaper and easier to separate. Additionally, the water formed as a byproduct, is
continuously removed and the equilibrium will continuously shift producing more
product until all of the reactants are consumed. The absorption of water by molecular
sieves or an azeotropic distillation with toluene could be both good options to remove
the water from the reaction mixture.

One the one hand, molecular sieve adsorbents are crystalline alumina-silicates, known
as zeolites. Their unique structure allows the water of crystallization to be removed,
leaving a porous crystalline structure. These pores or "cages" have a high affinity to re-
adsorb water or other polar molecules. Aided by strong ionic forces due to the
presence of cations such as sodium, calcium and potassium and by the absolutely
enormous internal surface area of close to 1000 m 2/g, molecular sieves will adsorb a
considerable amount of water or other fluids. If the fluid to be adsorbed is a polar
compound, it can be adsorbed with high loading, even at very low concentrations of
the fluid. Molecular sieves will therefore remove many gas or liquid impurities to very
low levels. Another feature of molecular sieve adsorbents is their ability to separate
gases or liquids by molecular size or polarity.

These crystals are characterised by a three-dimensional pore system, with identical

pores of precisely defined diameter. This structure is formed by tetrahedras of AlO 4
and SiO4. These tetrahedras are the basic building blocks for various zeolite structures,
such as zeolites A, the most common commercial adsorbent.

Due to the presence of alumina, zeolites exhibit a negatively charged framework,

which is counterbalanced by positive cations resulting in a strong electrostatic field on
the internal surface. These cations can be exchanged to adjust the pore size or the
adsorption characteristics. For instance, the sodium form of zeolite A has a pore
opening of approximately 4 Angstrom (4 x 10–10 m), called 4A molecular sieve. If the
sodium ion is exchanged with the larger potassium ion, the pore opening is reduced to
15
approximately 3 Angstrom, called 3A molecular sieve (Table 5 and 6). The ability to
adjust the pores to precisely determined uniform openings allows for molecules
smaller than its pore diameter to be adsorbed whilst excluding larger molecules, hence
the name “molecular sieve”. The different pore sizes of synthetic zeolites open up a
wide range of possibilities in terms of "sieving" molecules of different size or shape
from gases and liquids.

Table 5. Properties and characteristics of 3A molecular sieves [9].

Type 3A
Composition 0.6 K2O: 0.40 Na2O : 1 Al2O3 : 2.0 SiO2 : x H2O
Description The 3A form is made by substituting potassium cations for the
inherent sodium ions of the 4A structure, reducing the effective pore
size to 3 Å, excluding diameter > 3 Å
Properties 3 Å pore diameter, 8-12 mesh (common in liquid phase applications),
adsorbed water 19-20 % (w/w), regeneration temperature 175-260 ºC.
Major Commercial dehydration of unsaturated hydrocarbon streams,
applications including cracked gas, propylene, butadiene, acetylene; drying polar
liquids such as methanol and ethanol. Adsorption of molecules such as
NH3 and H2O from a N2/H2 flow. Considered a general-purpose drying
agent in polar and nonpolar media.

Table 6. Properties and characteristics of 4A molecular sieves [9].

Type 4A
Composition 1 Na2O : 1 Al2O3 : 2.0 SiO2 : x H2O
Description This sodium form represents the type A family of molecular sieves.
Effective pore opening is 4 Å, thus excluding molecules of effective
diameter >4 Å.
Properties 4 Å pore diameter, 8-12 mesh, adsorbed water 20-21 % (w/w),
regeneration temperature 200-315 ºC.
Major Preferred for static dehydration in closed liquid or gas systems, water
applications scavenging in printing and plastics systems and drying saturated
hydrocarbon streams. Adsorbed species include SO2, CO2, H2S, C2H4,
C2H6, and C3H6. Generally considered a universal drying agent in polar
and nonpolar media.

Molecular sieves (zeolites of types 3A and 4A) are widely used to dry laboratory
solvents. Dry organic solvents are important for many industrial sectors. Adsorptive
water removal is one technique to obtain highly pure solvents. It is widely used in the
drying, refining and purification of liquids and gases [10].

On the other hand, the equilibrium can be shifted towards products by removing the
water as an azeotrope. Water can form an azeotrope with certain alcohols at specific
concentrations. A special apparatus, called a Dean Stark apparatus or trap, can be used

16
to distil off the water as an azeotrope while the ester being is formed. Removing water
from the reaction in this way will favour ester formation.

Furthermore, another variation of an esterification is used to obtain the maximum

possible yield of the reaction, the Steglich Esterification. It is a variation of an
esterification with dicyclohexylcarbodiimide (DCC) as a coupling reagent and 4-N,N-
dimethyilaminopyridine (DMAP) as a catalyst. This reaction takes place at room
temperature and a suitable solvent could be acetonitrile.

Scheme 6. Steglich Esterification.

It is a mild reaction, which allows the conversion of sterically demanding and acid
labile substrates. It's one of the convenient methods for the formation of tert-butyl
esters because t-BuOH tends to form carbocations and isobutene after a subsequent
elimination under the conditions employed in the Fischer-Speier Esterification.

The application of the DCC method in pure organic synthesis dates back to 1967. The
Steglich Esterification proceeds according to the following general mechanism
(Scheme 7). Initially, the DCC and the carboxylic acid are able to form an O-acylisourea
intermediate, which offers reactivity similar to the corresponding carboxylic acid
anhydride. The alcohol may now add to the activated carboxylic acid to form the stable
diclyclohexylurea (DHU) and the ester [11].

17
Scheme 7. Reaction mechanism of the Steglich Esterification.

In peptide synthesis, the reaction with carboxylic acids, DCC and amines leads to
amides without problems (Scheme 8). This is because amines are more nucleophilic
than alcohols. Strong nucleophiles such as amines react readily with the O-acylisourea
and therefore need no additives such as DMAP.

Scheme 8. Fast reaction between amines and O-acylisourea.

18
In the case of alcohols, the esterification is slow (Scheme 9) and a side-reaction occurs,
diminishing the final yield or complicating purification of the product. This side-
reaction is a 1,3-rearrangement of the O-acyl intermediate to an N-acylurea which is
unable to further react with the alcohol (Scheme 10).

Scheme 9. Slow reaction between alcohols and O-acylisourea.

Scheme 10. Formation of N-acylurea through an acyl migration.

These drawbacks can be overcome by addition of catalytic amounts of p-

aminopyridines. For the efficient formation of esters, the addition of approximately 5
mol % DMAP is crucial. DMAP accelerates the reaction because, as a stronger
nucleophile than the alcohol, reacts with the O-acylisourea leading to a reactive amide
(active ester).

This intermediate cannot form intramolecular side products but reacts rapidly with
alcohols. DMAP suppresses the side reaction, acting as an acyl transfer reagent in this
way, and subsequent reaction with the alcohol gives the ester (Scheme 11).

19
Scheme 11. Reaction mechanism when the catalyst DMAP is used.

20
 6. MATERIALS AND METHODOLOGY
6.1. CHEMICAL PRODUCTS

The physical properties, hazards and safety of all chemical products used in this study
appear in Table 7.

Table 7. Physical properties, hazards and safety of the chemical products used [12].

Reactive Purity Bp MW Density Hazards Safety

(%) (ºC) (g/mol) (g/mL)
Acetic acid 99.9 118 60 1.05 H226: Flammable liquid and Use a
vapour fumehood
H312: Harmful in contact with
skin
H314: Causes severe skin burns
and eye damage
H318: Causes serious eye damage
H335: May cause respiratory
irritation

Isopentyl 99 129 88.2 0.81 H225: Highly Flammable liquid Avoid

alcohol and vapour contact
H226: Flammable liquid and with skin,
vapour eyes and
H315: Causes skin irritation clothing
H318: Causes serious eye damage
H332: Harmful if inhaled
H335: May cause respiratory
irritation
H336:May cause drowsiness or
dizziness

Sulfuric acid 96.5 337 98.1 1.84 H290: May be corrosive to metals Use a
H314: Causes severe skin burns fumehood
and eye damage
H318: Causes serious eye damage
H335: May cause respiratory
irritation

21
 p-toluene 99 38 190.22 1.24 H290: May be corrosive to metals Use a
sulfonic acid (m.p.) H302: Harmful if swallowed fumehood
monohydrate H314: Causes severe skin burns
and eye damage
H315: Causes skin irritation
H318: Causes serious eye damage
H319: Serious eye irritation
H335: May cause respiratory
irritation

Sodium Not 50 84 2.1 H319: Serious eye irritation Avoid

bicarbonate known (m.p.) contact
with eyes

Sodium 98 801 58.44 2.17 H319: Serious eye irritation Avoid

chloride (m.p.) contact
with eyes

Anhydrous Not 772 110.98 2.2 H319: Serious eye irritation Avoid
calcium known (m.p.) contact
chloride with eyes

Isopentyl Not 142 130.2 0.867 H226: Flammable liquid and Avoid
acetate known vapour contact
H315: Causes skin irritation with skin,
H319: Causes serious eye eyes and
irritation clothing
H335: May cause respiratory
irritation
H336: May cause drowsiness or
dizziness
H372: Causes damage to organs
through prolonged or repeated
exposure

Molecular Not - 202.1 - H315: Causes skin irritation Avoid

sieves known H319: Causes serious eye contact

22
 irritation with skin
H335: May cause respiratory and eyes
irritation

Toluene 99.5 110.6 92.14 0.86 H225: Highly Flammable liquid Avoid
and vapour contact
H304: May be fatal if swallowed with skin,
and enters airways eyes and
H315: Causes skin irritation clothing
H336: May cause drowsiness or
dizziness
H361: Suspected of damaging
fertility or the unborn child
H373: Causes damage to organs
through prolonged or repeated
exposure

Amberlyst-15 - - 314.4 - H319: Causes serious eye Avoid

irritation contact
with eyes

DMAP 98 110 122.2 - H301+H331: Toxic if swallowed or Avoid

(m.p.) if inhaled contact
H301: Toxic if swallowed with skin,
H310: Fatal in contact with skin eyes and
H311: Toxic in contact with skin clothing.
H314: Causes severe skin burns Wear
and eye damage nitrile
H315: Causes skin irritation gloves and
H318: Causes serious eye damage use
H319: Causes serious eye fumehood.
irritation
H331: Toxic if inhaled
H335: May cause respiratory
irritation
H411: Toxic to aquatic life with
long lasting effects

23
 DCC 99 34 206.3 - H302: Harmful if swallowed Avoid
(m.p.) H311: Toxic in contact with skin contact
H314: Causes severe skin burns with skin,
and eye damage eyes and
H317: May cause an allergic skin clothing.
reaction Wear
H318: Causes serious eye damage nitrile
H330: Fatal if inhaled gloves and
use
fumehood.

DCM 99.8 39.8 84.9 1.33 H302: Harmful if swallowed Avoid

H315: Causes skin irritation contact
H319: Causes serious eye with eyes
irritation and skin.
H335: May cause respiratory
irritation
H336: May cause drowsiness or
dizziness
H341: Suspected of causing
genetic defects
H351: Suspected of causing
cancer
H373: Causes damage to organs
through prolonged or repeated
exposure

DHU Not 230 224.3 - H302: Harmful if swallowed Use a

known (m.p.) fumehood

Acetonitrile 99.9 82 41.1 0.78 H225: Highly Flammable liquid Use a

and vapour fumehood
H302: Harmful if swallowed
H312: Harmful in contact with
skin
H319: Causes serious eye
irritation
H332: Harmful if inhaled

24
 6.2. MATERIAL

Different laboratory set-ups are used in order to do the different variations of the
reaction. First of all, a laboratory reflux apparatus for heating the chemical reaction is
used (Table 8).

Then, when the molecular sieves are used to absorb the water from the reaction
mixture, a cylindrical pressure-equalizing dropping funnel is used. The molecular sieves
are placed inside this glassware, in order to avoid the contact of these with the
reaction mixture and only have contact with the water. The side arm of the glassware
is isolated, so the vapours of water go up by this, then condense in the cooling system
above and condensing fluid gets into contact with the sieves.

Also a Dean-Stark trap is employed for the removal of water from the reaction mixture
in order to alter equilibrium concentrations. The reaction is carried out in a solvent
that forms an azeotrope with water, in this case toluene. The reaction mixture is
refluxed and the distillate condenses and drips into the trap, whereupon it separates
into two layers. The volume of condensate increases until the organic solvent drips
back into the reaction vessel, while the water remains in the trap. Thus, continual
azeotropic removal of water/toluene is effected with only a relatively small volume of
the organic solvent being required.

Finally, a Claisen fractional distillation head is used in order to separate a mixture of

components with similar boiling points.
Table 8. Differents laboratory set-ups with special glassware.

Cylindrical
Claisen fractional
Reflux pressure-equalizing Dean-Stark trap
distillation head
dropping funnel

25
 6.3. CHARACTERIZATION EQUIPMENT

Two techniques are used to confirm the presence of isoamyl acetate, nuclear magnetic
resonance spectroscopy (NMR) and infrared spectroscopy (IR).

 Spinsolve Education NMR-spectrometer (Table 9):

Table 9. NMR system data

Spectrometer type Spinsolve Education

Protocol 1D Proton
Spectral range 63 to -54 ppm
Scan type (number of scans) Standardscan (4 scans)
Aquisition time 1 minut
Linewidth at 50% peak height <0,7 Hz (for standard calibration 10% v/v
H2O/D2O sample)
Linewidth at 55% peak height <20 Hz (for standard calibration 10% v/v
H2O/D2O sample)
Field strength 1 Tesla
Proton Larmor frequency 43 Mhz

 Thermo Fisher IR200 IR-spectrometer, Serienummer AFN0600591

6.4. METHODOLOGY

The project consists in the synthesis of isoamyl acetate, a colourless liquid with
banana-like odour, by direct Fischer-Speier esterification of acetic acid and isoamyl
alcohol. The aim of the project is to make variations of the reaction to obtain the
maximum possible yield. In this way, the best reaction conditions for the synthesis of
isoamyl acetate will be known.

6.4.1. Sulfuric Acid Catalyst

In order to synthesize isoamyl acetate, using sulfuric acid as catalyst, the procedure
explained below is followed [13]. A flowchart can be found in Appendix A.

MODUS:

16 mL of isopentyl alcohol (0.147 mol) and 22 mL of glacial acetic (0.385 mol) are
added into a 100 mL Erlenmeyer. Carefully with swirling, 1 mL of concentrated sulfuric
acid (0.018 mol) is added to the contents of the flask, and also boiling stones or a
stirring bar. A reflux apparatus is assembled (Figure 1) and the reaction mixture is
heated at 100ºC reflux for 1 hour. Then the heat source is removed and the mixture
cooled to room temperature using a water bath of room-temperature water.

26
 The cooled mixture is poured into a separatory
funnel and 50 mL of cold distilled water are
added carefully. The reaction flask is rinsed
with 10 mL of cold water and this rinse is also
added to the separatory funnel. The funnel is
stoppered and inverted several times before
separating the lower aqueous layer from the
upper organic layer. The aqueous phase is set
aside.

25 mL of a 0.5M sodium bicarbonate solution

are poured into the separatory funnel. The
two phases are stirred until the evolution of
CO2 gas nearly stops. Then the funnel is
stopped, carefully is turned it upside down,
and immediately the CO2 gas that forms is
vented. The funnel is shaken and vented
frequently until no more gas evolved when the
Figure 1. Reflux apparatus.
funnel is vented. The lower aqueous layer is
removed and the extraction is repeated with another 25 mL of 0.5M sodium
bicarbonate solution. NaHCO3 extractions are continuously done until the lower layer
remains basic, using a litmus paper after the extraction. The organic layer is washed a
last time with 20 mL of 4M sodium chloride solution. The aqueous layer is removed
again and the ester is poured into a dry 50 mL Erlenmeyer flask.

The product is dried with anhydrous calcium chloride, allowing it to stand over the
drying agent for 10 minutes. The product is filtered from the CaCl2 through a small plug
of glass wool into a 50 mL round-bottomed flask. A simple distillation apparatus is
assembled using an oil-bath to heat. The fraction boiling between 134 and 145ºC is
collected in a tared 25 mL round-bottomed flask.

6.4.2. p-Toluenesulfonic Acid Catalyst

The first variation is the use of a different catalyst. Instead of sulfuric acid, p-
toluenesulfonic acid is used. This catalyst is a solid does not contain water, so by using
this catalyst, no water will be add to the reaction mixture that could influence the
equilibrium.

The procedure used to synthesize isoamyl acetate using p-toluenesulfonic acid as

catalyst is the same as using sulfuric acid. The only difference is that instead of adding
1 mL of sulfuric acid, a certain amount of p-toluenesulfonic acid is added and
dissolved. The amount of catalyst needed is known by calculating the concentration of
the sulfuric acid.

27
 96.5 𝑔 𝐻2 𝑆𝑂4 𝑚𝑜𝑙 𝐻2 𝑆𝑂4 1.84 𝑔 1000 𝑚𝐿
× × × = 18.1 𝑀
100 𝑔 98 𝑔 𝑚𝐿 𝐿

If 1 mL of sulfuric acid is used and the concentration is 18.1 M:

𝐿 18.1 𝑚𝑜𝑙
1 𝑚𝐿 × × = 0.018 𝑚𝑜𝑙
1000 𝑚𝐿 𝐿

𝑚𝑜𝑙 𝑝 − 𝑡𝑜𝑙𝑢𝑒𝑛𝑒𝑠𝑢𝑙𝑓𝑜𝑛𝑖𝑐 𝑎𝑐𝑖𝑑 𝑚𝑜𝑛𝑜ℎ𝑦𝑑𝑟𝑎𝑡𝑒 190.22 𝑔

0.018 𝑚𝑜𝑙 𝐻2 𝑆𝑂4 × × ×
𝑚𝑜𝑙 𝐻 + 𝑚𝑜𝑙

100
× = 3.46 𝑔 𝑝 − 𝑡𝑜𝑙𝑢𝑒𝑛𝑒𝑠𝑢𝑙𝑓𝑜𝑛𝑖𝑐 𝑎𝑐𝑖𝑑 𝑚𝑜𝑛𝑜ℎ𝑦𝑑𝑟𝑎𝑡𝑒
99

6.4.3. Amberlyst-15 Catalyst

The second variation is the use of a solid catalyst instead of the previous. The capacity
of Amberlyst-15 is 4.7 mmol H+/g by dry weight. The sulfuric acid (-3; 1.96 pKa) is
considered as a mono-protic acid because only the first proton is acidic enough to
acidify acetic acid (4.76 pKa). Compared to the 1 mL of acid sulfuric:

𝑚𝑜𝑙 𝑎𝑐𝑖𝑑𝑖𝑐 𝐻 +
0.018 𝑚𝑜𝑙 × = 0.018 𝑚𝑜𝑙 𝐻 + = 18 𝑚𝑚𝑜𝑙 𝐻 +
𝑚𝑜𝑙 𝐻2 𝑆𝑂4

𝑔 𝐴𝑚𝑏𝑒𝑟𝑙𝑦𝑠𝑡 − 15
18 𝑚𝑚𝑜𝑙 𝐻 + × = 3.83 𝑔 𝐴𝑚𝑏𝑒𝑟𝑙𝑦𝑠𝑡 − 15
4.7 𝑚𝑚𝑜𝑙 𝐻 +

This amount of Amberlyst-15 is added to the reaction mixture, not exceeding 120ºC. It
is removed at the end of the reaction through a filtration [17].

6.4.4. Molecular sieves

The next variation is the use of molecular sieves to

absorb the water of the reaction mixture, keeping other
parameters as the sulfuric acid catalyst constant. As well
as trying two types of molecular sieves (3A and 4A), is
also studied two ways of placing them, either in contact
with the reaction mixture or without contact.

For both types of molecular sieves (3A and 4A) is used

the same procedure. In the first case, using the “reflux
set-up” (first set-up of Table 8 and Figure 2), the
molecular sieves would be placed within the Erlenmeyer
with the reaction mixture. After the reaction, a filtration Figure 2. Set-up with molecular sieves
is needed to remove the molecular sieves. in contact with the reaction mixture.

28
 In the second case, the molecular sieves
would be placed without contact with the
reaction mixture, at a point between the
Erlenmeyer and the reflux, inside the
cylindrical pressure-equalizing dropping
funnel (second set-up of Table 8 and Figure
3). In this case, the filtration is not necessary.

In order to know the amount of molecular

sieves needed, it is calculated the amount of
water formed by the reaction.

Figure 3. Set-up with molecular sieves inside the

cylindrical pressure-equalizing dropping funnel.

0.81 𝑔 99 𝑔 𝑚𝑜𝑙 𝑚𝑜𝑙 𝐻2 𝑂 18 𝑔 1 𝑚𝐿

16 𝑚𝐿 𝑖𝑠𝑜𝑎𝑚𝑦𝑙𝑖𝑐 𝑎𝑙𝑐𝑜ℎ𝑜𝑙 × × × × × ×
𝑚𝐿 100 𝑔 88.2 𝑔 𝑚𝑜𝑙 𝑎𝑙𝑐𝑜ℎ𝑜𝑙 𝑚𝑜𝑙 1𝑔
= 2.6 𝑚𝐿 𝐻2 𝑂

Molecular sieves 3A and 4A have an adsorption water capacity of 20 % (w/w), and with
this information it can be known the amount of molecular sieves needed.

100 𝑔 𝑎𝑑𝑠𝑜𝑟𝑏𝑒𝑛𝑡
2.6 𝑔 𝐻2 𝑂 × = 13 𝑔 𝑎𝑑𝑠𝑜𝑟𝑏𝑒𝑛𝑡
20 𝑔 𝑤𝑎𝑡𝑒𝑟 𝑎𝑑𝑠𝑜𝑟𝑏𝑒𝑑

Good results are obtained in drying organic solvents with 20% (w/v) molecular sieves
[15], so the amount of molecular sieves is also calculated and studied by this way.

16 𝑚𝐿 𝑎𝑙𝑐𝑜ℎ𝑜𝑙 𝑖𝑠𝑜𝑎𝑚𝑦𝑙𝑖𝑐 + 22 𝑚𝐿 𝑎𝑐𝑒𝑡𝑖𝑐 𝑎𝑐𝑖𝑑 + 1 𝑚𝐿 𝑠𝑢𝑙𝑓𝑢𝑟𝑖𝑐 𝑎𝑐𝑖𝑑

= 39 𝑚𝐿 𝑠𝑡𝑎𝑟𝑡𝑖𝑛𝑔 𝑙𝑖𝑞𝑢𝑖𝑑

20 𝑔 𝑎𝑑𝑠𝑜𝑟𝑏𝑒𝑛𝑡
39 𝑚𝐿 × = 7.8 𝑔 𝑎𝑑𝑠𝑜𝑟𝑏𝑒𝑛𝑡
100 𝑚𝐿
6.4.5. Azeotropic distillation

Another variation is the use of a Dean-Stark trap for the removal of water from the
reaction mixture in order to alter equilibrium concentrations (third set-up of Table 8
and Figure 4). The reaction is carried out in a solvent that forms an azeotrope with
water, therefore in the reaction mixture is added 15 mL of toluene.

The boiling point of the azeotropic mixture of water and toluene is 84 ºC (b.p. toluene
110 ºC, b.p. water 100 ºC). The reaction mixture is refluxed and the distillate
condenses and drips into the trap, whereupon it separates into two layers. The volume
of condensate increases until the organic solvent drips back into the reaction vessel,

29
while the water remains in the trap. It can be observed approximately 2.6 mL of water
remained in the trap (Figure 5).

Figure 4. Dean-Stark set-up. Figure 5. Dean Stark trap with 2.6 mL of water
removed.
All the procedure is the same, except that in the distillation a Claisen fractional
distillation head is used (last set-up of Table 8). Not only to separate the mixture of
components with similar boiling points, but also to be sure that all the toluene is gone.

6.4.6. Steglich reaction

The last variation is the Steglich Esterification, used to obtain the maximum possible
yield of the reaction. This reaction is a variation of an esterification with
dicyclohexylcarbodiimide (DCC) as a coupling reagent and 4-N,N-
dimethyilaminopyridine (DMAP) as a catalyst.

In this esterification variation is followed a protocol with carboxylic acid as the limiting
reagent. In order to compare the yields, the amounts are adjusted as follows:

𝑙𝑖𝑚𝑖𝑡𝑖𝑛𝑔 𝑟𝑒𝑎𝑔𝑒𝑛𝑡: 0,147 𝑚𝑜𝑙 𝑖𝑠𝑜𝑝𝑒𝑛𝑡𝑦𝑙 𝑎𝑙𝑐𝑜ℎ𝑜𝑙 → 0,147 𝑚𝑜𝑙 𝑎𝑐𝑒𝑡𝑖𝑐 𝑎𝑐𝑖𝑑

The other reagents’ amounts are calculated using the molar ratio of the reaction
protocol followed (0.14 mmol of DMAP, 1.35 mmol of acid, 1.76 mmol of alcohol and
1.76 mmol of DCC) [16].
1.76 𝑚𝑚𝑜𝑙 𝑎𝑙𝑐𝑜ℎ𝑜𝑙
 0.147 𝑚𝑜𝑙 𝑎𝑐𝑒𝑡𝑖𝑐 𝑎𝑐𝑖𝑑 × = 0.192 𝑚𝑜𝑙 𝑖𝑠𝑜𝑝𝑒𝑛𝑡𝑦𝑙 𝑎𝑙𝑐𝑜ℎ𝑜𝑙
1.35 𝑚𝑚𝑜𝑙 𝑎𝑐𝑖𝑑

88.2 𝑔 100 𝑚𝐿
0.192 𝑚𝑜𝑙 𝑖𝑠𝑜𝑝𝑒𝑛𝑡𝑦𝑙 𝑎𝑙𝑐𝑜ℎ𝑜𝑙 × × × = 21 𝑚𝐿 𝑖𝑠𝑜𝑝𝑒𝑛𝑡𝑦𝑙 𝑎𝑙𝑐𝑜ℎ𝑜𝑙
𝑚𝑜𝑙 99 0.81 𝑔

30
 1.76 𝑚𝑚𝑜𝑙 𝐷𝐶𝐶
 0.147 𝑚𝑜𝑙 𝑎𝑐𝑒𝑡𝑖𝑐 𝑎𝑐𝑖𝑑 × 1.35 𝑚𝑚𝑜𝑙 𝑎𝑐𝑖𝑑 = 0.192 𝑚𝑜𝑙 𝐷𝐶𝐶
206.3 𝑔 100
0.192 𝑚𝑜𝑙 𝐷𝐶𝐶 × × = 40 𝑔 𝐷𝐶𝐶
𝑚𝑜𝑙 99

0.14 𝑚𝑚𝑜𝑙 𝐷𝑀𝐴𝑃

 0.147 𝑚𝑜𝑙 𝑎𝑐𝑒𝑡𝑖𝑐 𝑎𝑐𝑖𝑑 × = 0.015 𝑚𝑜𝑙 𝐷𝑀𝐴𝑃
1.35 𝑚𝑚𝑜𝑙 𝑎𝑐𝑖𝑑

122.2 𝑔 100
0.015 𝑚𝑜𝑙 𝐷𝑀𝐴𝑃 × × = 1.9 𝑔
𝑚𝑜𝑙 98

The procedure followed for this synthesis is explained below [16]:

MODUS:

To a solution of DMAP (0.015 mol) and acetic acid (0.147

mol) in acetonitrile (30 mL), a solution of isopentyl
alcohol (0.192 mol) in acetonitrile (80 mL) is added.
Then, a solution of DCC (0.192 mol) in acetonitrile (90
mL) is added dropwise to the reaction mixture at 0°C.
The reaction mixture is stirred during 3 hours at room
temperature (Figure 6), after which the acetonitrile is
evaporated in vacuo.

The residue is taken up in dichloromethane (DCM),

resulting in the formation of a white precipitate of
dicyclohexylurea. The suspension is filtered and the
solvent is evaporated in vacuo (Figure 7). Figure 4. Reaction mixture stirring at
room temperature.

Then, as in the previous procedures, the ester is

isolated by distillation, collecting the fraction boiling
between 134 and 145ºC.

The DCC and DMAP are highly toxic compounds,

therefore, it has to be worked with caution and very
safely. It is strictly important to work under
fumehood and using nitrile gloves. The nitrile gloves
show excellent resistance to most chemicals and
especially strong solvents, in addition the nitrile film
is highly resistant to punctures and tears.

Figure 5. Evaporation of solvent in vacuo.

31
 7. RESULTS AND DISCUSSION

The percentage yield of the reaction variations is compared in order to decide which
one gets the maximum amount of product possible.

Amounts of products calculated from the complete reaction of the limiting reagent is
called theoretical yield, whereas the amount actually produced of a product is the
actual yield. The ratio of actual yield to theoretical yield expressed in percentage is the
percentage yield.

𝑎𝑐𝑡𝑢𝑎𝑙 𝑦𝑖𝑒𝑙𝑑
𝑃𝑒𝑟𝑐𝑒𝑛𝑡𝑎𝑔𝑒 𝑌𝑖𝑒𝑙𝑑 = × 100
𝑡ℎ𝑒𝑜𝑟𝑒𝑡𝑖𝑐𝑎𝑙 𝑦𝑖𝑒𝑙𝑑

The theoretical yield is calculated from the limiting reagent:

0.81 𝑔 99 𝑚𝑜𝑙 𝑚𝑜𝑙 𝑖𝑠𝑜𝑝𝑒𝑛𝑡𝑦𝑙 𝑎𝑐𝑒𝑡𝑎𝑡𝑒

16 𝑚𝐿 𝑖𝑠𝑜𝑝𝑒𝑛𝑡𝑦𝑙 𝑎𝑙𝑐𝑜ℎ𝑜𝑙 × × × × ×
𝑚𝐿 100 88.2 𝑔 𝑚𝑜𝑙 𝑖𝑠𝑜𝑝𝑒𝑛𝑡𝑦𝑙 𝑎𝑙𝑐𝑜ℎ𝑜𝑙
130.2 𝑔
× = 18.9 𝑔
𝑚𝑜𝑙
𝑎𝑐𝑡𝑢𝑎𝑙 𝑦𝑖𝑒𝑙𝑑 𝑎𝑐𝑡𝑢𝑎𝑙 𝑦𝑖𝑒𝑙𝑑
𝑃𝑒𝑟𝑐𝑒𝑛𝑡𝑎𝑔𝑒 𝑌𝑖𝑒𝑙𝑑 = × 100 = × 100 = 𝑌𝑖𝑒𝑙𝑑 %
𝑡ℎ𝑒𝑜𝑟𝑒𝑡𝑖𝑐𝑎𝑙 𝑦𝑖𝑒𝑙𝑑 18.9

The yield Y is a function of reaction temperature, reaction time, type of catalyst,

reaction set-up, removal of water…

𝑌 = 𝑓(𝑡𝑒𝑚𝑝𝑒𝑟𝑎𝑡𝑢𝑟𝑒, 𝑡𝑖𝑚𝑒, 𝑐𝑎𝑡𝑎𝑙𝑦𝑠𝑡, 𝑠𝑒𝑡 − 𝑢𝑝, 𝑟𝑒𝑚𝑜𝑣𝑎𝑙 𝑜𝑓 𝑤𝑎𝑡𝑒𝑟 … )

When comparing different variations, it is strictly spoken necessary to change only one
parameter while keeping all other parameters constant. In this study, the reaction
temperature was set at 100ºC and the reaction time at 1 hour. Higher conversion
would be achieved by extending the reaction time, but 1 hour is enough to compare
the results.

In order to provide evidence that the results are reproducible, the whole experiment
independently was done twice. Therefore, the yield result is an average of 2
experiments. In all the experiments, the repetitions gave almost equal results.

7.1. CATALYST

First of all, three different acid catalysts were used to increase the rate of the reaction
and overcome the slow kinetics of the reaction. These catalysts were sulfuric acid, p-
toluenesulfonic acid and Amberlyst-15.

To compare the potential of Amberlyst-15 as acid catalyst with conventional catalysts,

the reaction yields are evaluated. The results are shown in Table 10. It can be observed

32
that the reaction with sulfuric acid catalyst ends up with a higher yield than both p-
toluenesulfonic acid (4% less) and Amberlyst-15 (13% less).

Table 10. Yield results using different catalysts.

Catalyst Percentage Yield

Sulfuric Acid Catalyst 79%
p-Toluenesulfonic Acid Catalyst 75%
Amberlyst-15 Catalyst 66%

A convenient way to express the relative strength of an acid is by the value of its pK a.

𝑝𝐾𝑎 = − log 𝐾𝑎

Small values of pKa equal large values of Ka (dissociation constant) and, as the pKa
decreases, the strength of the acid increases.

Certainly is that both sulfuric acid and p-toluenesulfonic acid are strong acids, but
there is a small difference in pKa between sulfuric acid (pKa of -3.0) and TsOH (pKa of -
2.8). So when comparing the results, it is possible that the lower acidity strength of
TsOH is the responsible of decreasing the conversion.

When comparing sulfuric acid with Amberlyst-15, a conventional heterogeneous

catalyst, the higher conversion can be explained by the initial homogeneous nature of
the hydrophilic sulfuric acid which reacts faster than heterogeneous hydrophobic
Amberlyst-15.

Heterogeneous acid catalysts suffer from severely decreased or even lost activity in the
presence of moisture due to water adsorption on the active sites. Also lower activity
may arise due to interaction of acid group with the support or due to more restricted
conformations of intermediate complexes adsorbed onto the solid surface in
comparison with homogeneous catalysts. [17] Moreover, the Amberlyst-15 was added
to the reaction mixture and in the end of the reaction it had to be removed through a
filtration. The filtration, in addition to that a part of the product was stuck to the solid
catalyst, decrease the yield.

Although higher conversion is achieved with sulfuric acid and p-toluenesulfonic acid
catalysts, their corrosive and hazardous nature and the subsequent need for
neutralization generating large quantities of aqueous waste restrict their industrial
applicability.

Due to these conventional catalysts’ disadvantages, solid acid catalysts have received
much attention as potential replacements for mineral liquid acids. The advantages of
heterogeneous solid acid catalysts include ease of separation from the reaction
medium, good recyclability, reduction of corrosion and enhanced product selectivity.

33
The isolation and reuse of this catalyst is an example of “Green Chemistry”, reducing
consumption of non-renewable resources.

Given these points, Amberlyst-15 is selected as the optimal catalyst since a high
conversion and high turnover frequency is highly desired for industrial applications.

7.2. REMOVAL OF WATER

Fischer-Speier esterification reaction takes advantage of Le Chatelier’s principle to

increase the amount of alcohol that is esterified. The equilibrium is shifted towards
products by decreasing the concentration of a product by removing it from the
equilibrium. This causes more reactants to be consumed to restore the product
concentration. Therefore, the water formed as a byproduct was continuously removed
by molecular sieves or by an azeotropic distillation with toluene.

In the first place, molecular sieves were added to the reaction mixture to adsorb
water. Two types of molecular sieves (3A and 4A) were used either in the reaction
mixture and also placed above the reaction mixture. Sulfuric acid and Amberlyst-15
were used as catalysts, but not TsOH since the result in the previous study was very
similar to sulfuric acid.

By using 13 g of adsorbent, amount calculated by the 20% (w/w) adsorption water

capacity, the yield obtained is 46%. As this result is very low and good results are
obtained in drying organic solvents with 20% (w/v) molecular sieves [15], was used 7.8
g of adsorbent instead of 13 g. The results are shown in Table 11.
Table 11. Yield results with 7.8 g of molecular sieves (3A and 4A) using different set-up and catalyst.

Percentage Yield
Set-up Catalyst
3A 4A
In contact with reaction Sulfuric Acid 72% 71%
mixture Amberlyst-15 49% 48%
Without contact with Sulfuric Acid 74% 72%
reaction mixture Amberlyst-15 65% 64%

First of all, it can be observed in all cases that the reaction with sulfuric acid catalyst
ends up with a higher yield than with Amberlyst-15, as expected from the previous
study.

To continue, there is almost no difference between the two types of molecular sieves,
but it is worth noting that in all cases 3A molecular sieves achieve between 1-2%
higher yield. 4A molecular sieve has a pore opening of approximately 4 Å (4 x 10–10 m),
but if the sodium ion is exchanged with the larger potassium ion, the pore opening is
reduced to approximately 3 Å (3A molecular sieve). Both molecular sieves adsorb

34
water (2.6 Å) because is a molecule smaller than its pore diameters, excluding larger
molecules. All other reactants and products are larger than 4 Å. The only one that
could have adsorption problems is the acetic acid, since its size is just over 4 Å. This
could explain the small improvement in the experiments with 3A molecular sieves.

In reference to the set-up, the reaction with molecular sieves in contact with the
reaction mixture ends up with a lower yield. This may be due to the fact that in this
case, a filtration after the reaction was needed to remove the molecular sieves from
the product. Also a lot of product was stuck to the solid, therefore lowering the yield.
This problem is observed when working in small quantities, at the laboratory level. But
on a large scale it is not so important, and for this reason at industrial level it can work
well.

Placing the molecular sieves above the reaction mixture ends up with a higher yield,
but it does not reach an improvement compared to the previous study, in which it was
not carried out the removal of water.

To further evaluate if the removal of water favour the ester formation, the water was
removed by an azeotropic distillation with toluene. The three different acid catalysts
were used. The results are shown in Table 12.
Table 12. Yield results with azeotropic distillation and different catalysts.

Catalyst Percentage Yield

Sulfuric Acid 85%
p-Toluenesulfonic Acid 79%
Amberlyst-15 71%

It can be observed, as expected, that the reaction with sulfuric acid catalyst ends up
with a higher yield than both p-toluenesulfonic acid (6% less) and Amberlyst-15 (14%
less).

In reference to the removal of water, a great improvement is observed comparing with

the first study. Higher conversion is achieved in all cases; a 6% higher yield with sulfuric
acid, 4% with TsOH and finally 5% with Amberlyst-15. Therefore, the use of a Dean-
Stark trap for the removal of water from the reaction mixture, in order to alter
equilibrium concentrations, is a good option to get the maximum amount of product
possible.

All things considered, it can be affirmed that the removal of water is a good option to
shift the equilibrium producing more product. The absorption of water by molecular
sieves or by an azeotropic distillation with toluene are both good options for the
removal of water.

35
On the one hand, molecular sieves are widely used in the drying, refining and
purification of liquids and gases. Also to obtain dry organic solvents which are
important for many industrial sectors. Most other drying agents, such as anhydrous
sodium sulphate or calcium chloride, would not remove water at the higher
temperatures used to make esters.

On the other hand, the azeotropic distillation with toluene is a technique often used
for the removal of water and the results obtained confirm it. Gaining 6% more
conversion in a large-scale reaction can greatly reduce economic costs and the
environmental impact.

7.3. STEGLICH ESTERIFICATION

The last variation is the Steglich Esterification, using carbodiimide activators to obtain
the maximum possible yield of the reaction. The yield obtained was the highest for the
normal set-up, 83%.

The use of DCC as a promoter represents one of the most versatile esterification
methods. Although this reagent is irritant to skin and a stoichiometric dosage or more
is necessary, this procedure enjoys various advantages. The reaction usually proceeds
at room temperature, and the reaction conditions are so mild that substrates with
various functional groups can be employed. The reaction is not sensitive to the steric
bulk of the reactants, allowing production of esters of tertiary alcohols. As such, a wide
range of applications has been achieved in the fields of natural products, peptides,
nucleotides, etc [14].

The favourable catalytic action of DMAP proves to be a useful agent in the study on
the esterification of carboxylic acids. Indeed, the addition of DMAP accelerates the
DCC-activated esterification of carboxylic acids with alcohols, such an extent that
formation of side products is suppressed and even sterically demanding esters are
formed in good yields at room temperature.

Steglich esterification has been used to facilitate the esterification of acids and
alcohols, allowing the conversion of sterically demanding substrates. The fact that this
reaction is not sensitive to the steric bulk of the reactants can explain the high
conversion obtained. Although the isoamyl alcohol is primary, it has an isopropyl group
that could present steric impediments.

It was no necessary to use the Chatelier’s Principle to shift the equilibrium towards
products, by increasing the concentration of the reactants or by decreasing the
concentration of a product. Therefore, the protocol allows high conversion without the
need of removing the water and with almost equimolar amounts of alcohol and acid.
Moreover, it is a mild reaction which allows the conversion of substrates without
requiring high temperatures.

36
 7.4. ISOAMYL ACETATE CHARACTERIZATION

The characterization of the isoamyl acetate was done by 1H NMR spectra. NMR results
obtained are exposed under Figure 8.

5’

2’ 4’
1’ 5’
3’
Figure 6. Isoamyl acetate structure.

1
H NMR (43 MHz, CDCl3): δ = 0.55 (6H, d, 5’CH3), δ = 0.98 (3H, m, 3’CH2 and 4’CH(CH3)3 ),
δ = 1.48 (3H, s, 1’CH3CO), δ = 3.61 (2H, t, 2’CH2O).

1
Figure 7. Isoamyl acetate H NMR spectra.

In figure 9 is shown the isoamyl acetate 1H NMR spectra. It can be observed that 0.55
ppm signal is a doublet that integrates six, so it fits very well with the six protons
bonded to 5’C due to the fact that they are coupled with the single proton in 4’C. These
protons are the furthest from the electronegative acetate group, so they are shielded
to a higher degree by a higher electron density of its surrounding molecular orbital.
Therefore, its NMR frequency will be shifted "upfield" with a lower chemical shift.

About the signal at 0.98 ppm, it can be said that is a multiplet that integrates three.
This is due to the fact that the proton bonded to 4'C and the two protons of 3'C have
very close signals and therefore overlap in the same peak, integrating three. It is a
complex signal (multiplet) because of the coupling with neighbouring protons.

37
The signal at 1.48 ppm is a singlet that integrates three, so it fits perfectly with the
three protons of the acyl group (1’CH3CO), that can not be coupled with any proton.

Finally, the signal at 3.61 ppm is a triplet that integrates two and corresponds with the
protons of 2’C. It is a triplet due to the fact that they are coupled with the two protons
bonded to 3’C. These protons are less shielded by such surrounding electron density
because they are next to an acetate group, an electronegative functional group that
attracts the electronic cloud. Therefore, its NMR frequency will be shifted "downfield"
with a higher chemical shift.

Appendix B contains an ampliation of the 1H NMR spectra (Figure 9), including the
spectra of isoamyl acetate analytical standard.

In addition, this characterization was supported by infrared spectroscopy. The most

important wavenumbers are 1055 cm-1 and 1210 cm-1 (νstret C-O), 1366 cm-1 (νbend CH3),
1466 cm-1 (νbend CH3 & CH2), 1739 cm-1 (νstret C=O), 2956 cm-1 (νstret CH3, CH2 & CH).

νbend CH3
& CH2
νstret CH3, CH2 & CH

νstret
νbend CH3
C=O νstret
C-O

Figure 8. Isoamyl acetate infrared spectra.

Appendix C contains an ampliation of the infrared spectra (Figure 10), including also
the spectra of isoamyl acetate analytical standard.

38
 8. CONCLUSIONS

Esterification will continue to play an important role both in organic synthesis and in
the chemical industry. However, further innovations need to be pursued in response to
the demands emerging from the continuing progress in organic synthesis.

The mail goal of this final thesis was to develop new methods to obtain the maximum
conversion of the isoamyl acetate synthesis. In this way, the best reaction conditions
for this shynthesis would be known. First of all, Amberlyst-15 is selected as the optimal
catalyst since a high conversion and high turnover frequency is highly desired for
industrial applications. High conversion is reached and the catalyst can be recycled and
reused. Special attention is given to the development of an efficient green
esterification process, although higher conversion is achieved with sulfuric acid and p-
toluenesulfonic acid catalysts.

Secondly, it is confirmed that the removal of water is a good option to shift the
equilibrium producing more product. The absorption of water by molecular sieves or
by an azeotropic distillation with toluene are both good options for the removal of
water. Particularly, the azeotropic distillation allows achieving high conversion, and
this fact, in a large-scale reaction can greatly reduce economic costs and the
environmental impact.

Finally, the use of carbodiimide activators through the Steglich Esterification is one of
the convenient methods for the formation of esters. This esterification variation
allows high conversion without the need of removing the water and with almost
equimolar amounts of alcohol and acid. Moreover, it is a mild reaction which allows
the conversion of substrates without requiring high temperatures.

It should be remembered that pot-treatment processes such as separation, isolation

and purification of the products and recovery of catalysts in practical processes often
take more time and energy than the reaction itself. Achievement of 100% yields with
equimolar amounts of reactants simplifies the separation steps and reduces excesses.

Not only esterification, but all chemical processes, should be as optimal as possible.
Since the materials involved in esterification (esters, carboxylic acids and alcohols) are
mostly non-toxic, attention has to be devoted to solvents. There are already some
processes that do no not use solvent, and it would be highly desirable to develop these
lines further. It can reasonably be said that esterification technology has the potential
to contribute a great deal to green chemistry.

39
 9. LIST OF REFERENCES

[1] Emil Fischer, Arthur Speier; “Darstellung der Ester”; Chemische Berichte, 1895, 28,
3252–3258.

[2] B. Neises, W. Steglich; Simple Method for the Esterification of Carboxylic

Acids; Angew. Chem. Int. Ed., 1978, 17 (7), 522-524.

[3] Pavia, Donald L.; Lampman, Gary M.; Kriz, George S.; Engel, Randall G.; Introduction
to Organic Laboratory Techniques: A Microscale Approach, 5th Edition; Brooks/Cole
Laboratory Series for Organic Chemistry, 2007; 109-117.

[4] Plata, C.; Millán, C.; Mauricio, J. .; Ortega, J., Formation of ethyl acetate and
isoamyl acetate by various species of wine yeasts; Food Microbiol. 2003, 20 (2), 217–
224.

[5] American Society of Enologists., C. J. van; Augustyn, O. P. H.; Wet, P. de; Joubert,
W. A. American journal of enology and viticulture; American Society of Enologists,
1979; Vol. 30.

[6] NIST Chemistry Webbook https://webbook.nist.gov/chemistry/ (Date consulted 27

May 2018)

[7] Sigma-Aldrich; Amberlyst-15 hydrogen form, Technical Information Bulletin;

https://www.sigmaaldrich.com/catalog/product/sial/216380?lang=es&region=ES
(Date consulted 26 April 2018).

[8] Pal, Rammohan; Sarkar, Taradas; Khasnobis, Shampa; Amberlyst-15 in organic

synthesis; ARKIVOC. 2012, (i) 570-609.

[9] Sigma-Aldrich; Molecular Sieves, Technical Information Bulletin;

https://www.sigmaaldrich.com/chemistry/chemical-synthesis/learning-
center/technical-bulletins/al-1430/molecular-sieves.html (Date consulted 13 March
2018).

[10] Pahl, C.; Pasel, C.; Luckas, M.; Bathen, D. J., Adsorptive Water Removal from
Primary Alcohols and Acetic Acid Esters in the ppm-Region; Chem. Eng. Data 2012, 57
(9), 2465–2471.

[11] Organic Chemistry Portal; Steglich Esterification; http://www.organic-

chemistry.org/namedreactions/steglich-esterification.shtm (Date consulted 17 April
2018)

[12] Pub Chem, Open Chemistry Database; Chemical and Physical Properties;
https://pubchem.ncbi.nlm.nih.gov/search/search.cgi (Date consulted 3 March 2018)

40
[13] R. Mohrig, Jerry; Noring Hammond, Christina; et al; Moderns Projects and
Experiments in Organic Chemistry: Miniscale and Standard Taper Microscale, 2nd
edition; W.H. Freeman and Company: New York, 2003; 93-96.

[14] Otera, Junzo; Esterification: Methods, Reactions and Applications, Wiley-VCH:

Weinheim, 2003, 15.

[15] Williams, D. B. G.; Lawton, M. J.; Drying of Organic Solvents: Quantitative

Evaluation of the Efficiency of Several Desiccants; Org. Chem. 2010, 75 (24), 8351–
8354.

[16] B. Neises, W. Steglich; Simple Method for the Esterification of Carboxylic

Acids; Angew. Chem. Int. Ed., 1978, 17 (7), 522-524.

[17] Verheyen, T.; Smet, M.; De Borggraeve, W. M.; Water Tolerant and Reusable
Sulfonated Hyperbranched Poly(aryleneoxindole) Acid Catalyst for Solvent-Free
Esterification; ChemistrySelect, 2017, 2, 1-8.

41
 REAGENTS
APPENDIX A 16 mL isopentyl alcohol
22 mL glacial acetic acid
1 mL sulfuric acid/catalyst

REACTION
Heat the reaction mixture at
reflux for 1h

SEPARATION
Separatory funnel with 50 mL
of cold distilled water

AQUEOUS LAYER
ORGANIC LAYER

AQUEOUS LAYER ORGANIC LAYER

Dry with anhydrous

calcium chloride, 10 min

FILTRATION

SIMPLE DISTILLATION
42
Collect the fraction
boiling between 135-145
C
APPENDIX B
ISOAMYL ACETATE ANALYTICAL STANDARD 1H NMR SPECTRA

43
ISOAMYL ACETATE 1H NMR SPECTRA

44
APPENDIX C
ISOAMYL ACETATE ANALYTICAL STANDARD INFRARED SPECTRA

45
ISOAMYL ACETATE INFRARED SPECTRA

46