CLASSIFICATION TESTS FOR CARBOXYLIC ACIDS AND THEIR DERIVATIVES

Cabalo, A.*, Cachuela, B., Casapao, C., Daza, C., de Belen, G.

Department of Pharmacy, Faculty of Pharmacy
University of Santo Tomas

ABSTRACT
Carboxylic acids and their derivatives are organic compounds that bears an acyl group
that may be attached to an electronegative atom such as oxygen, nitrogen or sulfur, or a
substituent (-Y) that can act as the leaving group in nucleophilic acyl substituion (S N Acyl). In SN
Acyl, carboxylic acids and their acid derivatives would act as an electrophile and would be
attacked by a nucleophile. All carboxylic acid derivatives if subjected to hydrolysis would
produce their corresponding parent carboxylic acid. Among all the acid derivatives acyl halides
are the most reactive and amides are the least. The objective of this experiment is to distinguish
the different carboxylic acids and their derivatives and to classify them according to their
reactivities. Classification tests such as hydrolysis, alcoholysis, aminolysis and hydroxamic acid
test was performed on acetyl chloride, acetic anhydride, ethyl acetate, acetamide and
benzamide. The results has shown that positive results such as the odor, precipitate formed and
evolution of a gas in hydrolysis would determine the presence of their carboxylic acids. This is
true for acetyl chloride, acetic anhydride, ethyl acetate and benzamide. While the odor of the
resulting mixture in alcoholysis proves the presence of the ester that was formed upon addition
of ethanol to acetic acid, acetyl chloride and acetic anhydride. Formation of a white precipitate in
aminolysis of acetyl chloride and acetic anhydride is a positive result of the presence of an
anilide which is acetanilide. Hydroxamic Acid Test is used to identify the presence of the
functional group of esters and amides. A positive result of red-purple colored solution was seen
in ethyl acetate and acetamide. Overall, determining the reactivities of carboxylic acids and their
derivatives can be obtained by performing different classification tests. This is made possible
due to the difference in their substituents and the reagents used.
____________________________________________________________________________
in rancid odor of sour butter and hexanoic
INTRODUCTION

acid in aroma of goats.

Carboxylic Acid is a class of organic

compound that bears a carboxyl group
Figure 1. Structure of a Carboxylic Acid

(COOH). The carboxyl group is named as

such because of the presence of a carbonyl
group (C=O) and a hydroxyl group (-OH).
The chief and most obvious characteristic of
carboxylic acids is their acidity. They react
with bases such as sodium hydroxide to
produce metal carboxylate salts, RCO2-M+.
Many carboxylic acids are found in nature
such as acetic acid in vinegar, butanoic acid

Carboxylic

Acid

derivatives

are

compounds that bears an acyl group

(RC=O) bonded to an electronegative atom
(O, N or S) or substituent (-Y) which is the
leaving

group

in

nucleophilic

acyl

substitution (SN Acyl). There are many acid

derivatives known but the most common

ones are the acid halides, acid anhydrides,

undergo basic hydrolysis but in esters it is

esters and amines.

also known as saponification.

In a nucleophilic acyl substitution, an

addition of a nucleophile initially forms a
tertahedral intermediate that eliminates one

Figure 4. General Mechanism of Acid Halide Hydrolysis

of the two substituents (the leaving group)

originally bonded to the carbonyl carbon.

Figure 5. General Mechanism of Ester Hydrolysis

Figure 2. General Mechanism of SN Acyl

All

acid

derivatives

can

be

Acid halides are the most reactive

hydrolyzed to yield a carboxylic acid but the

carboxylic acid derivatives. They can be

conditions required range from mild to

converted

severe.

into

many

other

kinds

of

compounds by nucleophilic acyl substitution

Alcoholysis reactions are usually

mechanism.

carried out in the presence of pyridine or

sodium hydroxide to react with the hydrogen
chloride formed. Acid halides and acid
anhydrides react with an alcohol to yield an
ester. A primary alcohol would react faster to
Figure 3. Interconversions of carboxylic acid derivatives

Hydrolysis is a typical SN Acyl

an acid halide or acid anhydride than a

secondary or tertiary alcohol.

process and is intiated by the attack of

water. The hydroxide ion would attack the
carbonyl

group

to

give

tetrahedral

Figure 6. General Mechanism of Alcoholysis

intermediate and the expulsion of the

substituent would follow to produce the

Aminolysis is a chemical reaction in

carboxylic acid and a by-product. Hydrolysis

which a compound is reacted with ammonia

of acid halides and acid anhydrides can

or an amine. The reaction of acid halides

undergo under normal conditions since they

and acid anhydrides to an ammonia or

are both very reactive. Esters and amides

amine will yield an amide.

collected. The samples used are acetyl

chloride, acetic anhydride, ethyl acetate,
acetamide and benzamide.
Figure 7. General Mechanism of Acid Halide Aminolysis

I.

Hydrolysis of Acid Derivatives

Hydroxamic Acid is a class of

A. Acyl

organic compound bearing the functional

group

RC(O)N(OH)R1 which is

halides

and

Acid

anhydrides

usually

Using the samples acetyl

prepared from esters, acid chlorides or

or

chloride and acetic anhydride, 10

Hydroxamate Test is a chemical test that

drops were cautiously added to the

detects the presence of the functional

test tube filled with 1 mL of water. A

groups of an ester and an amide. A positive

warming effect was noted. The

result of red-purple color shows that an

resulting mixture was then divided

ester or an amide has been converted into a

into two portions. To the first portion,

hydroxamic acid.

1 mL of

carboxylic

acids.

Hydroxamic

Acid

2% AgNO3 was added.

Formation

of

precipitate

was

observed. To the second portion, 1

mL

of

saturated

NaHCO3

was

added. An evolution of a gas was

Figure 8. Functional Group of a Hydroxamic Acid

observed.
B. Esters
The sample used for the

Figure 9. Structure of a Hydroxamic Acid

The objectives of this experiment is

hydrolysis of esters is ethyl acetate.

the

In a test tube of 1 mL ethyl acetate,

carboxylic acids and their derivatives and to

2 mL of 25% NaOH solution was

distinguish

added. The mouth of the test tube

to

differentiate

the

them

reactivities
using

of

different

was covered with a marble. The test

classification tests.

tube was then heated in a boiling

METHODOLOGY

water bath for 5 minutes. The

The materials and equipments were

resulting mixture was neutralized

prepared and cleaned prior to the start of

with 10% HCl solution and with a

the

wafting motion, the odor was noted.

experiment

to

prevent

possible

contamination. Reagents and samples were

C. Amides

The sample used for the

added

to

each

test

tube.

The

hydrolysis of amides is benzamide.

mixtures were shaken for several

In a test tube, 1 mL of benzamide

minutes. The odor and layers of

was treated with 5 mL of 10% NaOH

ester formed was observed.

solution. The resulting mixture was

heated until it boils. A piece of red

III.

litmus paper was placed on the

Anhydrides

mouth of the test tube to test the

reaction of gas evolved. The litmus
papers

change

of

color

was

observed.
II.

Aminolysis : Anilide Formation

Acyl
halides
and
Acid

Prior to the test 5 mL of water

was placed into two different test tubes.
In one test tube of water, a mixture of 23 drops acetyl chloride and 0.5 mL

Alcoholysis : Schotten-Baumann

aniline was added and to the second

Reaction

test tube of water, a mixture of 2-3 drops

A. Acetic Acid

acetic anhydride and 0.5 mL aniline was

added. The precipitate formed was

In a test tube, a mixture of 10

observed.

drops of acetic acid, 1 mL ethanol

and 5 drops of concentrated H2SO4

IV.

Hydroxamic Acid Test

was placed in a water bath for two

This test was performed on ethyl

minutes. The odor of the ester

acetate and acetamide. The samples did

formed was observed.

not undergo a preliminary test before the

B. Acyl

halides

and

Acid

test was conducted. In a mixture of 2 mL

anhydrides

of alcoholic NH2OH HCl and 1 mL of 1M

Two mixtures were prepared

KOH, 2 drops of ethyl acetate was

prior to the test and the samples

added. In another mixture of 2 mL of

used were acetyl chloride and acetic

anhydride. To the first test tube, 0.5
mL ethanol, 1 mL of water and 0.2
ml of acetyl chloride were mixed. To
the

second

test

tube,

0.5

mL

ethanol, 1 mL of water and 0.2 mL of

acetic anhydride were mixed. Two
milliliters of 20% NaOH solution was

alcoholic NH2OH HCl and 1 mL of 1M

KOH, 2 drops of acetamide was added.
The two resulting mixtures were heated
in a boiling water bath for two minutes.
After heating, the mixtures were cooled.
One milliliter of 5% FeCl3 was added to

each mixture. The color of the resulting

effervescence or the evolution of carbon

solution was observed.

dioxide. Both acetyl chloride and acetic

anhydride

RESULTS AND DISCUSSION

showed

bubbling

effect

therefore both samples yielded a carboxylic

Table 1. Hydrolysis of Acyl halides and Acid anhydrides

Sample

Warming
effect

Upon
addition
of
2%
AgNO3

Upon
addition
of
saturate
d
NaHCO3

Acetyl
Chloride

Warm
(+)

White
ppt (+)

Bubbling
effect (+)

Acetic
Anhydrid
e

Room
Temp (-)

Turbid
solution

Bubbling
effect (+)

acid upon hydrolysis.

Figure 10. Hydrolysis of Acyl Halide and Acid Anhydride

Test Results
Table 2. Hydrolysis of Esters

Sample

Observations

Ethyl Acetate

Plastic balloon odor

According to Table 1, acetyl chloride

had a warming effect when added to water

Ethyl acetate is the ester of acetic

while acetic anhydride showed none. Upon

acid and ethanol. In the experiment, since a

addition of 2% AgNO3, acetyl chloride

base (NaOH) was used to hydrolyze the

showed a formation of a white precipitate

sample a carboxylic acid salt was formed

while a turbid solution in acetic anhydride.

but the salt was easily converted into a

The turbid solution of acetic anhydride may

carboxylic acid with the addition of the HCl.

be due to the fact that is reacts slower than

Ethyl acetates original fruity odor became a

acetyl chloride.

plastic balloon-like odor due to the formation

The warming effect present was due

of the acetic acid and ethanol.

to the formation of acetic acid. This

formation

underwent

an

exothermal

reaction. Acetic anhydride did not show a

warming effect that may be due to possible
contamination or undesired experimental
errors.

Figure 11. Hydrolysis of Esters Test Results

The addition of NaHCO 3 determines

the presence of a carboxylic acid. A positive
result

would

show

the

formation

of

Table 3. Hydrolysis of Amides

Sample

Observations

Benzamide

Red to blue litmus

paper

After treating benzamide with NaOH

and upon heating, the ammonia gas is given
off which can be proven by the change of
color of the litmus paper from red to blue.

Figure 12. Hydrolysis

of Amides

Figure 13. Litmus

Paper Results

Figure 14. Alcoholysis of Acetic Acid Test Results

Table 5. Alcoholysis of Acyl halides and Acid anhydrides

Sample

Odor

Formation of
layers

Acetyl
Chloride

Fruity

None

Acetic
Anhydride

Fruity

None

Acetyl chloride and acetic anhydride

reacts with ethanol to yield the ester, ethyl
ethanoate. The difference between the two

Table 4. Alcoholysis of Acetic Acid

Sample

Observations

Acetic Acid

Plastic balloon odor

is that they produced different by-products.

Acetyl chloride would give off the hydrogen
chloride gas while acetic anhydride would

Alcoholysis

or

esterification

and

hydrolysis are reverse reactions. When

acetic acid is subjected to ethanol and

produce ethanoic acid. The fruity odor

produced was due to the ester formed since
esters generally has sweet pleasant smells.

concentrated sulfuric acid it produced the

ester, ethyl acetate. Concentrated sulfuric
acid was needed as an acid catalyst
because the reaction is slow and reversible.
Concentrated sulfuric acid was used to
reduce the chances of the reverse reaction
happening. The plastic balloon odor of the
resulting mixture was due to the formation
of ethyl acetate. The reverse reaction is
found on table 2.

Figure 15. Alcoholysis of Acyl Halides and Acid

Anhydrides Test Results

Table 6. Aminolysis of Acyl halides and Acid anhydrides

Sample

Appearance
of Precipitate

Upon
additiom
water

Acetyl
Chloride

White ppt

dissolved

Acetic
Anhydride

No ppt

(-)

solution

of
A positive result is exhibited by a
red-purple color solution. According to Table
7, both ethyl acetate and acetamide turned
into a brown turbid solution upon addition of
5%

FeCl3.

The

solution

was

actually

burgundy in color which is a shade of

Anilides are amides in which the

reddish brown. Brown turbid solution was an

hydrogen of the amido group is replaced by

incorrect description of the actual result.

phenyl. The white precipitate in acetyl

This was due to insufficient knowledge of

chloride

of

describing colors. Therefore both ethyl

acetanilide. Acetic anhydride should have

acetate and acetamide showed a positive

shown a white precipitate for the reason that

result.

is

due

to

the

formation

it will also produce acetanilide when reacted

to aniline. A possible contamination must
have lead to the incorrect result. Acetanilide
has a water solubility of 6.93 x 103
milligrams per liter at 77o Fahrenheit or 25o
Celsius. Acetanilide is slightly soluble in
water at room temperature that is why upon
addition of water the precipitate dissolved.

Figure 17. Hydroxamic Acid Test Results

CONCLUSION
It can be concluded that several
classification tests can be conducted to
differentiate the reactions of carboxylic acids
and their derivatives. Nucleophilic acyl

Figure 16. Aminolysis of Acyl Halids and Acid Anhydrides

Test Results
Table 7. Hydroxamic Acid Test

Sample

Upon addition of 5%
FeCl3

Ethyl Acetate

Brown
solution

turbid

Acetamide

Brown

turbid

substitution is the main mechanism of which

hydrolysis,

alcoholysis

and

aminolysis

undergoes.
Hydrolysis of acetyl chloride, acetic
anhydride, ethyl acetate and benzamide
showed

positive

results

to

prove

the

presence of their carboxylic acid and their

by-product/s. Alcoholysis of acetic acid,

Retrieved from November 26, 2016, from

acetyl chloride and acetic anhydride showed

http://www.vanderbilt.edu/AnS/Chemistr
y/Rizzo/Chem220b/Ch20.pdf

positive results to prove the presence of

their ester. Aminolysis of acetyl chloride
showed a positive result to prove the
presence of the formation of anilide. Postive
results in hydroxamic acid test on ethyl
acetate and acetamide proves the presence
of their functional group. Inaccurate results
may

have

been

due

to

possible

contamination and error in performing the

experiment.
Overall

the

experiment

was

success in achieving its objectives.

REFERENCES
[1] Chemical Tests for Unknowns. (n.d.).
Retrieved November 26, 2016, from
http://cactus.dixie.edu/smblack/chemlabs/ch
emical_tests_for_unknowns.pdf

[8] Aminolysis. (July 2016). Retrieved

November
26,
2016,
from
https://en.wikipedia.org/wiki/Aminolysis
[9] Hydrolysing Amides. (2004). Retrieved
November
28,
2016,
from
http://www.chemguide.co.uk/organicprops/a
mides/hydrolysis.html
[10] Carboxylic Acid. (2016) Retrieved
Novermber
28,
2016
from
https://www.britannica.com/science/carboxyl
ic-acid/Synthesis-of-carboxylicacids#ref999793
[11] Ethyl Acetate. (2016). Retrieved
November
28,
2016,
from
https://en.wikipedia.org/wiki/Ethyl_acetate
[12] The Mechanism for the Esterification
Reaction. (2002). Retrieved November 28,
2016,
from
http://www.chemguide.co.uk/physical/cataly
sis/esterify.html

[2] Identification of Unknowns. (n.d.).

Retrieved November 26, 2016, from
http://www.umsl.edu/~orglab/experiments/U
NKEXP.html

[13] The Reaction of Acid Anhydrides with

Water, Alcohols and Phenol. (2004).
Retrieved November 28, 2016, from
http://www.chemguide.co.uk/organicprops/a
nhydrides/oxygen.html

[3] Hydroxamic Acid (Nov. 16, 2015).

Retrieved November 26, 2016, from
https://en.wikipedia.org/wiki/Hydroxamic_aci
d

[14] Bathan, G., Bayquen, A., Crisostomo,

A., et al. (2014). Laboratory Manual in
Organic Chemistry (Revised Edition).
Quezon City : C & E Publishing Inc.

[4] McMurry, J. (2016). Organic Chemistry.

Singapore: Cengage Learning Asia Pte Ltd.

[15] Phenylamine as a Primary Amine.

(2004). Retrieved November 28, 2016, from
http://www.chemguide.co.uk/organicprops/a
niline/amine.html

[5] Carboxylic Acid. (March 29, 2016).

Retrieved November 26, 2016, from
https://www.britannica.com/science/carboxyl
ic-acid
[6] Carboxylic Acid Derivatives. (n.d.).
Retrieved November 26, 2016, from
http://crab.rutgers.edu/~alroche/Ch21.pdf
[7]
Carboxylic
Acid
Derivatives
:
Nucleophilic Acyl Substitution. (n.d.).

[16] Anilide. (n.d.) Retrieved November 28,

2016,
from
http://www.merriamwebster.com/medical/anilide
[17] What is the solubility of acetanilide?
(n.d.). Retrieved November 28, 2016, from
https://www.reference.com/science/solubility
-acetanilide-2ffd508a52eda2e8