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CHEM 200L EXPT # 10 2FMT GROUP # 4

CLASSIFICATION TESTS FOR CARBOXYLIC ACIDS AND ITS DERIVATIVES

Gregorio, A., Inocencio, P., Latayan, S., Laurente, T.*, Lazaro, C., Mangabat, R.
Department of Medical Technology, Faculty of Pharmacy
University of Santo Tomas

ABSTRACT

Carboxylic acids are organic compounds which contain at least one carboxyl group
(C(=O)OH). Some of its derivatives that were used in this experiment are: acyl halides, acid
anhydrides, esters, and amines, and the different sample compounds that were used for each
derivative are: acetyl chloride, acetic anhydride, ethyl acetate, acetamide, and benzamide
respectively. The different classification tests that were performed on the individual compounds
are: hydrolysis, alcoholysis (Schotten-Baumann reaction), aminolysis, and hydroxamic acid test.
These were done in order to determine if there is a carboxylic acid and/or its derivative present.
The objectives of this experiment are: 1. To differentiate the reactivities of carboxylic acid and its
derivatives. 2. To distinguish carboxylic acids and their derivatives using the different
classification tests listed beforehand. 3. To explain through chemical equations and mechanisms
the reactions that were involved in each test.

INTRODUCTION

Carboxylic acids are generally weak most biological pathways and many
acids that contain a carbonyl group (C=O) industrial processes. For example, acyl
which is attached to a hydroxyl group (-OH), halides are usually used as a starting
these two together is also known as a material for synthesis due to its high
carboxyl group. These derivatives have a reactivity. Acid anhydrides are commonly
carbonyl group attached to either a used in the preparation of acetate esters,
substituent or an electronegative element aspirin and a precursor to various resins.
which would determine the reactivity of the Amides, along with acids and esters, are
compound. The most common of its abundant in living organisms proteins,
derivatives, which were used in the nucleic acids and many pharmaceutical
experiment, are acyl halides (RCOX), acid drugs have amide functional groups.
anhydrides (RC(O))2O, esters (RCOOR),
and amides (RCONH2). Majority of
carboxylic acids derivatives are involved in
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All Carboxylic IUPAC Name Chemical

carboxylic acids Acid Structure
Derivative
derivatives yield the
parent carboxylic Acyl Halide Acetyl Chloride
(C2H3OCl)
acid upon reaction
with water.
Moreover, more
Acid Acetic Anhydride
reactive acid Anhydrides (C4H6O3)
derivatives easily
react to be
Esters Ethyl Acetate
converted to less (C4H8O2
reactive
derivatives. Their
Amides Acetamide
reactivity in (C2H5NO)
decreasing order is
as follows: acyl
halides, acid anhydrides, esters, amides,
and carboxylate ions. Nucleophilic acyl
substitution is the reaction mechanism that
Figure 2. Nucleophilic acyl substitution in
basic condition.

is involved with carboxylic acid to produce

its derivative. It is where a nucleophile
attacks the carbonyl carbon, forming an
unstable tetrahedral intermediate, which
Table 1. Carboxylic acids common
modifies a carboxylic acid derivative by
derivatives with proper nomenclature and
eliminating the weakest base. Confirmatory
chemical structure.
tests for carboxylic acid derivatives include:
Hydrolysis, alcoholysis (Schotten-Baumann
reaction), aminolysis, and hydroxamic acid
METHODOLOGY
test.
Figure 1. Nucleophilic acyl substitution in A. Hydrolysis of Acid Derivatives
acidic condition.

Acid Halides and Acid Anhydrides
1 mL of distilled water was placed inside
a test tube, then 10 drops of the sample
was added dropwise. The resulting mixture
was was divided into two portions then was
placed in two separate test tubes. The first
test tube was added with 2% Silver nitrate,
while 1 mL of saturated sodium bicarbonate
was added to the second test tube.
Formation of precipitate and evolution of
gas was noted.
Esters
2 mL of 25% sodium hydroxide solution
was added to 1 mL. The mouth of the test
tube was covered with a marble and was
allowed to heat in a boiling water bath for 2
minutes. The remaining mixture was
acidified with 10% hydrochloric acid
solution. The test tube was wafted to take
note of the odor.
Amides
Benzamide was treated with 5 mL of
10% sodium hydroxide solution and was
immersed in a boiling water bath. During
heating a moist red litmus paper was placed
near the mouth of the test tube.
B. Alcoholysis; Schotten-Baumann
Reaction
10 drops of glacial acetic acid was
placed inside a test tube. 1 mL of ethanol
and 5 drops of concentrated sulfuric acid
was warmed in a water bath for 2 minutes,
3
the odor was taken note of. While in another
test tube, a mixture of 0.5mL of ethanol, 1
mL of water, and 0.2 mL 20% sodium
hydroxide solution was added. The test tube
was covered with parafilm and was shaken
for several minutes. The formation of two
layers was taken note of.
C. Aminolysis: Anilide formation
0.5 mL of aniline was placed inside two
test tubes and 3 drops of acyl halide and
acetic anhydride was added dropwise into
the two test tubes respectively and was
mixed. The mixtures were transferred into
two new test tubes with 5 mL of water.
Formation of precipitate was noted.
D. Hydroxamic Acid Test
A preliminary test was done by mixing 1
mL of 95% ethanol and 1 M hydrochloric
acid to separate test tubes that contained 2
drops of the samples (ethyl acetate and
acetamide) to be used. If upon addition of 1
drop of 5% ferric chloride and there was no
yellow solution produced then another test
is conducted. 2 ml of alcoholic
hydroxylamine and 1 ml of 1M potassium
hydroxide was added to the separate test
tubes. The test tubes were then immersed
in a water bath for 5 minutes, then the
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mixtures were allowed to cool. After

cooling, 1 ml %% of ferric chloride was
added and the color of the formed
precipitate was noted.

RESULTS AND DISCUSSIONS Figure 3. Results of Hydrolysis of acetyl

A. Hydrolysis chloride and acetic anhydride

Table 2. Results observed from the

hydrolysis of acyl halides and acid
anhydrides

Sample +H20 +2% +Saturated CH 3 COCl+ H 2 O CH 3 COOH + HCl

AgNO3 NaHCO3
C 4 H 6 O 3+ H 2O 2 CH 3 COOH
Acetyl Warming White Evolution of
Chloride effect ppt. gas
Figure 4. Reaction mechanism of hydrolysis
of acetyl chloride and acetic anhydride
Acetic Warming Evolution of
Anhydride effect - gas
Upon adding 2% AgNO3, a white
precipitate was formed in the first test tube
A warming effect was observed in because AgCl(s) was formed. Acetic
the hydrolysis of both acetyl chloride and anhydride did not yield a precipitate due to
acetic anhydride due to the exothermal the absence of a halide which would react
reaction and formation of the parent to Ag+ to form a silver halide. Effervescence
carboxylic acid, acetic acid. was due to the evolution of CO2 gas as
indicated by the formation of bubbles when
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saturated NaHCO3 was added to the Table 4. Results observed from the
hydrolysis of amide
second test tube.
Sample +10% NaOH w/ heating

Table 3. Results observed from the Benzamide Red litmus turned blue
hydrolysis of esters

Sample +25% NaOH w/ heating

The hydrolysis of an ester in a basic for 5 mins, +10% HCl
medium, such as in 25% NaOH, will Ethyl acetate Vinegar-like odor
undergo a process called saponification. It
is where an ester is converted in to the
carboxylate salt of its parent carboxylic acid
Figure 6. Results observed from the
and an alcohol from which the ester was
hydrolysis of benzamide.
formed. The resulting solution was acidified
with 10% HCL, yielding acetic acid, the The hydrolysis of amides involves the same
esters parent carboxylic acid. mechanism as the other carboxylic acid
derivatives, which is nucleophilic acyl
'
RCOO R + NaOH RCOONa+ R ' OH substation, that yielded ethanoic acid and
an amine (NH3). The Red litmus paper
RCOONa+ HCl RCOOH + NaCl
turned to blue due to the fact that there is
NH3 present, which is a Lewis base.
C 4 H 8 O 2+ NaOH CH 3 COONa+ CH 3 CH 2 OH

CH 3 COONa+ HCl CH 3 COOH + NaCl

RCON R ' 2+ NaOH RCOONa+ R 2' NH
Figure 5. Reaction mechanism of
Hydrolysis of an ester in a basic medium.
RCOONa+ H RCOOH + Na

Figure 6. Reaction mechanism of

Hydrolysis of amides

B. Alcoholysis, Schotten-Baumann
Reaction
Table 5. Results observed from the
alcoholysis of acyl halide and acid
anhydride.
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Sample +Ethanol,+H2O, 20% Acetyl chloride Fruity Odor, formation

NaOH of 2 layers
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Acetic acid Fruity odor; formation of Table 6. Results observed from the
2 layers aminolysis of acyl halide and acid
anhydride.
Ethyl acetate was produced via
Sample +aniline,+ H2O
Fischer Esterification with the use of glacial
acetic acid by nucleophilic acyl substitution Acetyl chloride White ppt.

of ethanol. Heating the alcohol and the acid Acetic anhydride White ppt.
in the presence of an acid catalyst, in the
form of conc. H2SO4, causes ester
formation also known as esterification, Both acetyl chloride and acetic
along with dehydration. anhydride that underwent aminolysis with
Alcoholysis reactions are usually aniline (an amine) produced a white
carried out with the presence of NaOH, so precipitate indicating the formation of
that it would react with HCl to prevent any anilide, upon the addition of water. The
other unnecessary reactions. appearance of white precipitate in acetic
' '
RCOX+ R OH RCOO R + HX anhydride took more time than in acetyl
chloride due to the fact that the leaving
' '' ''
RCOOCO R + R OH RCOO R + R ' COOH group reactivity of the former is weaker than
the latter.
Sample Preliminary Upon
test addition of D. Hydroxamic
5% FeCl3
Ethyl acetate Yellow solution Deep Acid Test
burgundy
solution
Table 7. Results
Acetamide Yellow solution Deep
burgundy observed from the
solution hydroxamic acid test
CH 3 COCl+ CH 3 CH 2 OH C 4 H 8 O2+ HCl
of esters and amides.

C 4 H 6 O 3+CH 3 CH 2 OH C 4 H 8 O 2+CH 3 COOH A preliminary test was done prior to

the confirmatory test to eliminate the
Figure 7. Reaction mechanism of
alcoholysis of acetyl chloride and acetic phenols and enols that give color with ferric
acid. chloride in an acidic solution, that may give
a false positive result in the ferric

C. Aminolysis, anilide formation hydroxamate test. During this preliminary

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test, both samples yielded a yellow color

indicating that it is now safe to proceed to
the proper confirmatory test. REFERENCES
During the proper confirmatory test,
only ethyl acetate produced a deep McMurry, J. 2010. Fundamentals of Organic
burgundy color because it is an ester. Chemistry. Brooks Cole Publishing
Esters react with basic hydroxylamine to Company.Australia
produce a hydroxamic acid, which will react
with acidic FeCl3 to form the magenta Vogel, A. I., & Furniss, B. S. (1996). Vogel's
colored ferric hydroxamate. textbook of practical organic
chemistry. Edinburgh: Longman.

H. (2015). Definition of Saponification.

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