Experiment 9

Classification Tests for Hydroxyl- & Carbonyl- Containing Compounds

Sunglao, A., Supan, E., Tan, C., Tayag, P., Tuason, A.
Group # 9, 2G Medical Technology, Faculty of Pharmacy, University of Santo
Tomas

Abstract
In organic chemistry, functional groups are specific groups of atoms within molecules that are
responsible for the characteristic chemical reactions of those molecules and examples of functional
groups are hydroxyl and carbonyl groups. Hydroxyl groups have one hydrogen paired with one oxygen
atom (symbolized as OH) and are usually seen in alcohols while carbonyl groups have one oxygen atom
double-bonded to a carbon atom (symbolized as C=O) and are usually seen in aldehydes and ketones. In
this experiment, several differentiating tests were conducted with the samples ethanol, n-butyl alcohol,
sec-butyl alcohol, tert-butyl alcohol, benzyl alcohol, n-butyraldehyde, benzaldehyde, acetone,
acetophenone, isopropyl alcohol and acetaldehyde. These tests include solubility of alcohols in water,
Lucas test, Chromic Acid test (Jones oxidation), 2,4-Dinitrophenylhydrazone (2,4-DNP/ 2,4-DNPH) test,
Fehlings test, Tollens Silver Mirror test and Iodoform test. A positive result on solubility was seen on
alcohols under six (6) carbon atoms while the Lucas test differentiates 1, 2 and 3 alcohols, gives a
positive result of turbidity (alkyl chloride formation), and the rate of the reaction was observed. The
Chromic Acid test/Jones Test tests for oxidizable or any compounds that possess reducing property (has
an alpha acidic hydrogen) and 1, 2 and 3 alcohols and aldehydes give a positive visible result of a bluegreen suspension. On the other hand, the 2,4-Dinitrophenylhydrazone (2,4-DNP/2,4-DNPH) test is used
as confirmatory for carbonyl groups and therefore, gives a positive result of red-orange precipitate or
yellow precipitate for aldehydes and ketones. Fehlings test and Tollens Silver Mirror test are both used
to identify aldehydes wherein a positive result of brick-red precipitate can be seen through the Fehlings
test and silver mirror for the Tollens test. Iodoform test is used as confirmatory for methyl carbinol (2
alcohol with adjacent methyl group) and methyl carbonyl groups and shows a positive result of yellow
crystals or precipitate. In conclusion, n-butyraldehyde, benzaldehyde and acetaldehyde are aldehydes
while acetone and acetophenone are ketones and all of these contain carbonyl groups. Ethanol, n-butyl
alcohol, sec-butyl alcohol, tert-butyl alcohol and benzyl alcohol are all alcohols containing hydroxyl
groups wherein 3 alcohols are the most stable while 1 alcohols are the most polar among the three.

Introduction
An alcohol is a compound that has a
hydroxyl group bonded to a saturated, sphybridized carbon atom, R-OH (Figure 1).
Alcohols are classified as primary (1),
secondary (2), or tertiary (3), depending on the

number of carbon substituents bonded to the

hydroxyl-bearing carbon (McMurry, 2010). The
most important physical property of alcohols is
the polarity of their OH groups. Because of the
large difference in electronegativity between
oxygen and carbon and between oxygen and
hydrogen, both the C-O and O-H bonds of an

alcohol are polar covalent, and alcohols are

polar molecules (Brown & Poon, 2011).
Alcohols usually have much higher boiling
points than might be expected from their molar
masses. This difference can be understood if we
consider the types of intermolecular attractions
that occur in these liquids (Zumdahl & Zumdahl,
2012).

carbocation, which in turn reacts with the

chloride ion (nucleophile) to generate an alkyl
halide product. Figure 4 shows the general
mechanism for this SN1 reaction.

Figure 1. Structure of Alcohol

The functional group of an aldehyde is a
carbonyl group bonded to a hydrogen atom
(Figure 2). The functional group of a ketone is a
carbonyl group bonded to two carbon atoms
(Figure 3). Because of the polarity of the
carbonyl group, aldehydes and ketones are polar
compounds and interact in the liquid state by
dipole-dipole interactions. As a result, aldehydes
and ketones have higher boiling points than
those of nonpolar compounds with comparable
molecular weight (Brown & Poon, 2011).

Figure 4. General Mechanism of Lucas test

The Chromic acid test (Jones oxidation)
distinguishes primary and secondary alcohols
from tertiary. Chromic acid will oxidize a
primary alcohol first to an aldehyde and then to
a carboxylic acid and it will oxidize a secondary
alcohol to a ketone. Tertiary alcohols do not
react. The OH-bearing carbon must have a
hydrogen atom attached. Since the carbon atom
is being oxidized in primary and secondary, the
orange chromium Cr6+ ion is being reduced to
the blue-green Cr3+ ion. The reactions involved
are shown in Figure 5.

Figure 2. Structure of Aldehyde

Figure 3. Structure of Ketone

The Lucas reagent is an aqueous
solution of strong acid (HCl) and zinc chloride
(ZnCl). The alcohol starting material must be
sufficiently soluble in aqueous environments for
the reaction to take place. The reaction that
occurs in the Lucas test is an S N1 nucleophilic
substitution. The acid catalyst activates the OH
group of the alcohol by protonating the oxygen
atom. The C-OH+ bond breaks to generate the

Figure 5. General Mechanism of Jones

Oxidation
The 2,4-Dinitrophenylhydrazone test
serves as a derivative formation. Both aldehydes
and
ketones
react
with
2,4dinitrophenylhydrazine to form a solid 2,4dinitrophenylhydrazone (DNP) derivative. The
color of this derivative can also provide useful
structural information. If the solid is yellow, this
most often means that the carbonyl group in the

unknown is unconjugated. A reddish-orange

color most likely means that the carbonyl group
is conjugated. In a few cases, compounds in
which the carbonyl group is not conjugated
produce orange precipitates. Simply having a
double bon or phenyl group somewhere in an
aldehyde or ketone does not necessarily mean
that the carbonyl group is conjugated. The
double bond must be separated from the
carbonyl by one single bond only. If the double
bond is further away, it is isolated from the
carbonyl and not conjugated with the carbonyl.
Figure 6 illustrates the reaction occurring in the
2,4-DNP/2,4-DNPH test.

oxidized, whereas ketones are not. Tollens test

uses a reagent known as Tollens reagent, which
is a colorless, basic, aqueous solution containing
silver ions coordinated to ammonia [Ag(NH 3)2+].
Tollens reagent oxidizes an aldehyde into the
corresponding carboxylic acid. The reaction is
accompanied by the reduction of silver ions in
Tollens reagent into metallic silver, which, if the
test is carried out in a clean glass test tube,
forms a mirror on the test tube. Ketones are not
oxidized by the Tollens reagent, so the
treatment of a ketone with Tollens reagent in a
glass test tube does not result in a silver mirror.
The reactions involved in the silver mirror test
are shown in Figure 8.

Figure 8. General Mechanism of Tollens

Silver Mirror test
Figure 6. General Mechanism of 2,4Dinitrophenylhydrazone test
In Fehlings test, the presence of
aldehydes and not ketones is detected by
reduction of the deep blue solution of copper (II)
to muddy green solution, and then form a brickred precipitate of insoluble cuprous oxide
(Cu2O). This test is commonly used for reducing
sugars but is known to be not specific for
aldehydes. Figure 7 shows the general
mechanism undergone in the Fehlings test.

Figure 7. General Mechanism of Fehlings

test
Tollens test, also known as silver-mirror
test, is a qualitative laboratory test used to
distinguish between and aldehyde and a ketone.
It exploits the fact that aldehydes are readily

The Iodoform test indicates the presence

of an aldehyde or ketone in which one of the
groups directly attached to the carbonyl carbon
is a methyl group. Such a ketone is called a
methyl ketone. In the Iodoform test, the sample
is allowed to react with a mixture of iodine and
base. Hydrogens alpha to a carbony group are
acidic and will react with base to form the anion,
which then reacts with iodine in this way to
form the triiodo compound, which the reacts
with more base to form the carboxylic acid salt
plus iodoform, a yellow precipitate. Formation
of a yellow precipitate therefore indicates the
presence of a methyl group directly attached to
the carbonyl. The mechanism of the iodoform
reaction is that of alpha-halogenation of a
carbonyl compound under basic conditions,
followed by nucleophilic displacement of the
resulting triiodomethyl group by hydroxide.
Figure 9 illustrates the reaction involved in the
Iodoform test.

Figure 9. Genral Mechanism of Iodoform test

One of the objectives of this experiment
is to distinguish whether a compound is
hydroxyl- or carbonyl-containing. Other
objectives include differentiating the three types
of alcohols, differentiating aldehydes from
ketones, and explaining the mechanisms
involved in the differentiating tests.
Materials and Methods
A. Materials
In this experiment, the materials needed are
Lucas reagent, chromic acid reagent, 95%
ethanol, Fehlings A and B, Tollens reagent, 5%
NaOCl solution, iodoform test reagent, 2,4dinitrophenylhydrazine, Pasteur pipette, test
tubes, beaker, and the sample compounds
ethanol, n-butyl alcohol, sec-butyl alcohol, tertbutyl alcohol, benzyl alcohol, n-butyraldehyde,
benzaldehyde, acetone, acetophenone, isopropyl
alcohol and acetaldehyde.
B. Methods
1. Solubility of Alcohols in Water
In the five test tubes, 10 drops each of
ethanol, n-butyl alcohol, sec-butyl alcohol, tertbutyl alcohol and benzyl alcohol were placed
using a Pasteur pipette. 1 mL of water was added
dropwise in each of the test tubes containing
alcohol while shaking the mixture after each
addition. When cloudiness resulted, 0.25 mL of
water was added at a time, with vigorous
shaking, until a homogenous dispersion
appeared. The total volume of water added was
noted. When no cloudiness resulted after the
addition of 2.0 mL water, alcohol was noted
down to be soluble in water.
2. Lucas test

Three test tubes were prepared and 1 mL of

Lucas reagent was dropped in each of the test
tubes. On the first test tube, 2-3 drops of n-butyl
alcohol was added. The test tube was shook
vigorously for a few seconds and the mixture
was allowed to stand at room temperature. The
two other test tubes were placed with 2-3 drops
of sec-butyl alcohol and tert-butyl alcohol,
respectively, shook vigorously and the mixtures
were allowed to stand. The rate of formation of
cloudy suspension or the formation of two layers
was observed.
3. Chromic Acid test (Jones Oxidation)
Using the samples n-butyl alcohol, sec-butyl
alcohol, tert-butyl alcohol, n-butyraldehyde,
benzaldehyde, acetone, and acetophenone, one
drop of each liquid or small amount of the solid
sample was dissolved in 1 mL of acetone in
different test tubes. 2 drops of 10% aqueous
K2Cr2O7 solution and 5 drops of 6 M H 2SO4
were added in each of the test tubes.
4. 2,4-Dinitrophenylhydrazone
DNP/2,4-DNPH) test

(2,4-

One drop of a liquid sample (acetone,

acetaldehyde, n-butylraldehyde, benzaldehyde
and acetophenone) was placed in a test tube and
5 drops of 95% ethanol were added. After
shaking
well,
3
drops
of
2,4dinitrophenylhydrazine were added. The
solution was allowed to stand for at least 15
minutes when no yellow or orange-red
precipitate forms.
5. Fehlings test
Into each test tube, 1 mL of freshly prepared
Fehlings reagent (made by mixing equal
amounts of Fehlings A and Fehlings B) was
placed. 3 drops of the sample to be tested were
added and the test tubes were placed in a beaker
of boiling water. Changes were observed that
occurred within 10-15 minutes. The test was
performed on acetaldehyde, n-butyraldehyde,
acetone, benzaldehyde and acetophenone.
6. Tollens Silver Mirror test

Five test tubes with 1 mL each of freshly

prepared Tollens reagent were prepared. In each
of the test tubes, 2 drops of each of the samples
(acetaldehyde, benzaldehyde, acetone, nbutyraldehyde and acetophenone) were added
separately. The mixtures were shaken and
allowed to stand for 10 minutes. When no
reaction occurred, the test tubes were placed in a
beaker with warm water for 5 minutes.
Observations were recorded.
7. Iodoform test
2 drops of each sample (acetaldehyde, acetone,
acetophenone, benzaldehyde and isopropyl
alcohol) were placed in different test tubes. 20
drops of 10% KI solution were added. While
shaking, 20 drops of fresh chlorine bleach (5%
sodium hypochlorite) were added slowly to each
tube and mixed. The formation of a yellow
precipitate was noted.
Results and Discussion
For the test for solubility of alcohols in
water, turbidity of the solution was to be
observed. Cloudiness of the solution indicated
insolubility of that specific alcohol to water. The
amount of water needed to produce homogenous
dispersion was also observed. Table 1 shows the
data gathered from the test.
Table 1. Solubility of Alcohols in Water
Alcohol

ethanol
n-butyl
alcohol
sec-butyl
alcohol
tert-butyl
alcohol
benzyl
alcohol

Amount of
water (in
mL) needed
to produce a
homogenous
dispersion
1 mL
1.50 mL

Solubility to
water

1 mL

soluble

1 mL

soluble

2 mL

insoluble

soluble
soluble

As indicated in the table, only benzyl

alcohol was insoluble in water, while ethanol, nbutyl alcohol, sec-butyl alcohol and tert-butyl
alcohol were all soluble in water. This follows
the principle like dissolves like and therefore,
it can be said that the alcohols that were soluble
in water are polar compounds since water is
polar. Of the alcohols that were soluble in water,
ethanol, sec-butyl alcohol and tert-butyl alcohol
all required only 1 mL of water to be added in
order to be considered soluble. This indicates
that there are certain factors affecting solubility.
One of these is the presence of number of
carbon atoms. The lower the number of carbon
atoms present, the more soluble or more
miscible a substance is. Branching of carbon
chains also affect solubility. The more branching
present, the more soluble a compound is. This is
only true for organic compounds that have the
same number of carbon atoms present.
The Lucas test differentiated 1, 2 and
3 alcohols. Alkyl chloride formation was
observed and caused turbidity or cloudiness. The
rate of reaction was also observed. Table 2
presents the results of the Lucas test.
Table 2. Lucas test
Sample
n-butyl alcohol
sec-butyl alcohol
tert-butyl alcohol

Reaction observed
colorless
slightly turbid
turbid

According to the table above, n-butyl

alcohol was soluble in Lucas reagent while secbutyl alcohol and tert-butyl alcohol were
observed to have a formation of cloudy layer.
Tert-butyl alcohol took the shortest time to form
the layer while sec-butyl alcohol took the
longest time. The reaction mechanism involved
in the Lucas test is based on S N1 reaction, which
depends on the formation of stable carbocations.
Reactivity of alcohols in SN1 reaction is 3 > 2
> 1. 3 alcohols formed the second layer in less
than a minute. 2 alcohols required 5-10 minutes
while 1 alcohols were usually unreactive. The
presence of ZnCl2, a good Lewis acid, made the
reaction mixture even more acidic; thus, it
enhanced the formation of carbocations.

The Chromic Acid test (Jones

Oxidation) tested for oxidizables or any
compounds that possess reducing property (has
an alpha acidic hydrogen). Table 3 shows the
results gathered from the said test.
Table 3. Chromic Acid test (Jones Oxidation)
Sample
n-butyl alcohol
sec-butyl alcohol
tert-butyl alcohol
n-butyraldehyde
benzaldehyde
acetone
acetophenone

Reaction observed
blue-green solution
blue-green solution
orange solution
blue-green solution
blue-green solution
orange solution
orange solution

According to Table 3, n-butyl alcohol,

sec-butyl
alcohol,
n-butyraldehyde
and
bezaldehyde gave a positive result of blue-green
solution while tert-butyl alcohol, acetone and
acetophenone gave a result of orange solution.
Chromic Acid test / Jones Oxidation involved
reduction-oxidation or redox reaction. 1 and 2
alcohols and aldehydes underwent oxidation and
chromium underwent reduction from Cr 6+ to
Cr3+. 1 and 2 alcohols and aldehydes reduced
the orange-red chromic acid/sulphuric acid
reagent to an opaque green or blue suspension of
Cr (III) salts in 2-5 seconds. 1 alcohols reacted
with chromic acid to yield aldehydes, which are
further oxidized to carboxylic acids. 2 alcohols
reacted with chromic acid to yield ketones,
which do not oxidize further. 3 alcohols were
usually unreactive and aldehydes were oxidized
to carboxylic acids.
The 2,4-Dinitrophenylhydrazone (2,4DNP/2,4-DNPH) test detected the presence of
carbonyl groups and tests positive for aldehydes
and ketones. Table 4 shows the results from the
test.
Table 4. 2,4-Ditrophenylhydrazone (2,4DNP/2,4-DNPH) test
Sample
acetaldehyde
n-butyraldehyde
benzaldehyde

Reaction observed
yellow ppt
yellow ppt
yellow ppt

acetone
acetophenone

yellow ppt
red-orange ppt

As indicated in the table, only

acetophenone gave a result of red-orange
precipitate while the rest of the samples gave a
result of yellow precipitate. A result of redorange precipitate indicated the presence of
conjugated carbonyl compounds while a result
of yellow precipitate indicateed the presence of
unconjugated carbonyl compounds. The reaction
of 2,4-DNPH with aldehydes and ketones in an
acidic solution is a dependable and sensitive test.
Its reaction mechanism involved condensation or
nucleophilic addition of NH2 to C=O and
elimination of H2O. Some high molecular
weight ketones may fail to react or may yield
oils. Most aromatic aldehydes and ketones
produce red dinitrophenylhydrazone while many
nonaromatic aldehydes and ketones produced
yellow products.
Fehlings
test
was
another
differentiating test for aldehydes and ketones. In
this test, aldehydes gave a positive result of
brick-red precipitate while ketones did not
produce any reaction. Table 5 presents the
results of the test.
Table 5. Fehlings test
Sample
acetaldehyde
n-butyraldehyde
benzaldehyde
acetone
acetophenone

Reaction observed
brick-red ppt
brick-red ppt
brick-red ppt
blue solution
blue solution

As shown in the given table, only

acetone and acetophenone did not react to form
a precipitate while the rest gave a positive result
of brick-red precipitate. Fehlings test involved
reduction-oxidation
or
redox
reaction.
Aldehydes were oxidized to carboxylic acids
while ketones did not undergo oxidation. In
here, copper was reduced from Cu2+ to Cu1+.
Tollens Silver Mirror test differentiated
aldehydes from ketones wherein aldehydes were

expected to be oxidized while ketones did not

undergo any oxidation. Table 6 shows the results
from the said test.
Table 6. Tollens Silver Mirror test
Sample
acetaldehyde
n-butyraldehyde
benzaldehyde
acetone
acetophenone

Result observed
silver mirror
flesh solution
light yellow solution
with globules
dark-gray solution
turbid gray solution

According to Table 6, only acetaldehyde

formed a silver mirror. The samples nbutyraldehyde and benzaldehyde, although they
are aldehydes, did not form any silver mirror.
The ketones acetone and acetophenone formed
dark-gray solution and turbid gray solution,
respectively. The preparation of Tollens reagent
was based on the formation of a silver diamine
complex that is water soluble in basic solution.
The Tollens Silver Mirror test involved
reduction-oxidation
or
redox
reaction.
Aldehydes were oxidized to carboxylic acids
while ketones did not undergo oxidation except
alpha-hydroxyketone. Silver was reduced from
Ag1+ to Ag0.
Iodoform test was used to detect the
presence of methyl carbinol (2 alcohol with
adjacent methyl group) and methyl carbonyl
groups. Table 7 shows the results from the said
test.
Table 7. Iodoform test
Sample
acetaldehyde
n-butyraldehyde
benzaldehyde
acetone
acetophenone
isopropyl alcohol

Reaction observed
yellow ppt
no reaction
red ppt with globules
yellow ppt
yellow ppt
yellow crystal ppt

According
to
the
give
table,
acetaldehyde, acetone and acetophenone gave
aresult of yellow precipitate. Benzaldehyde gave

a result of red precipitate with globules while

isopropyl alcohol gave a result of yellow crystal
precipitates. No reaction was observed from nbutyraldehyde. In this test, yellow crystals or
precipitate gave a positive result. An alkaline
solution of sodium hypoiodite, formed from
sodium hydroxide and iodine, converted
acetaldehyde and aliphatic methyl ketones into
iodoform (haloform reaction). Since the reagent
was also an oxidizing agent, alcohols which are
readily oxidized to acetaldehydes or methyl
ketones also gave a positive reaction. The
mechanism of iodoform synthesis occurred
through a series of enolate anions, which are
iodinated; hydroxide displaced the Cl3- anion
through an addition/elimination pathway.
References
From books:
Brown, W., Poon, T. (2011). Introduction to
organic chemistry international student version
(5th edition). NJ, USA: John Wiley & Sons, Inc.
McMurry, J. (2010). Foundations of organic
chemistry (Philippine edition). USA: Cengage
Learning Asia Pte. Ltd.
Zumdahl, S., Zumdahl, S. (2012). Chemistry: An
Atoms First Appraoch (International edition).
USA: Brooks/Cole, Cengage Learning.
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