Classifying Compounds containing Hydroxyl- and Carbonyl groups using specified tests

Abraham, K.D., Acebedo, D.M.A., Alap-ap, M.L.M., Asprec, W.A.A., Balaaldia, K.C.A. and
Basilio, Z.A.S.
2B-PH, Group No. 1, Department of Pharmacy, Faculty of Pharmacy, University of Santo
Tomas, Espaa Boulevard, 1015 Manila, Philippines
ABSTRACT
Hydroxyl- or carbonyl- containing samples was given for this experiment for analysis. Hydroxyl
group refers to a functional group containing OH- when it is a substituent in an organic
compound whereas carbonyl group refers to a divalent chemical unit consisting of a carbon and
an oxygen atom connected by a double bond [1]. Hydroxyl group is the characteristic functional
group of alcohols and phenols while carbonyl group is the characteristic functional group of
aldehydes and ketones [2]. In this experiment, several differentiating tests were conducted with
samples ethanol, n-butyl alcohol, sec-butyl alcohol, tert-butyl alcohol, benzyl alcohol, nbutyraldehyde, benzaldehyde, acetone, acetophenone, isopropyl alcohol, and acetaldehyde.
These tests include solubility of alcohols in water, Lucas test, Chromic Acid test, 2,4Dinitrophenylhydrazone test, Fehlings test, Tollens Silver Mirror test, and Iodoform test. The
first three test done helped in the identification of the structure of the alcohols and tell if they are
Primary (the alpha carbon atom where OH group is attach is attached to only one alkyl group),
secondary (the alpha carbon is attached to two alkyl group), or tertiary (the alpha carbon is
attached to three alkyl groups). The fourth test, The 2,4-Dinitrophenylhydrazon test was used to
identify the presence of carbonyl groups-, aldehydes and ketones. 2,4-Dinitrophenylhydrazone
was also used to distinguish its aromaticity. A positive result of red-orange precipitate indicates a
carbonyl group, while a yellow precipitate indicates a presence of aldehydes and ketones. The
fifth and sixth test, the Fehlings and Tollens Silver Mirror test were used to identify the
presence of an aldehyde and distinguishing an aldehyde from a ketone. In this experiment, only
an aldehyde would yield a positive result while ketones wont. And lastly, the Iodoform test was
used to ensure the presence of CH3CO group or methyl carbonyl group in the samples given.
INTRODUCTION
An alcohol is a compound that has a
hydroxyl group bonded to a sp3-hybridized
carbon atom, R-OH. They are also classified
as primary (1), secondary (2) or tertiary
(3), depending on the number of carbon
substituents bonded to the hydroxyl-bearing
carbon. Their most important physical
property is the polarity of their OH groups.
Due to the large difference in the
electronegativity of C-O and O-H, both of
their bond of an alcohol are polar covalent
and alcohols are polar molecules. Hydroxyl
group is used to describe the functional

group OH when it is a substituent in an

organic compound. Hydroxyl groups are
known for their tendency to form hydrogen
bonds either as a donor or as an acceptor.
This is also related to their ability to increase
hydrophilicity and water solubility [3].
The functional group of an aldehyde is a
carbonyl group bonded to a hydrogen atom.
Another carbonyl group is a ketone bonded
to two carbon atoms. Due to the polarity of a
carbonyl group, aldehydes and ketones are
polar compounds and interact in the liquid
state by dipole-dipole interaction. The result
is that aldehyde and ketones have higher

boiling points than those of nonpolar

compounds with comparable molecular
weight [4].
The Lucas test is performed with the use of
the Lucas Reagent, where Lucas reagent is a
solution of anhydrous zinc chloride in
concentrated hydrochloric acid. This reagent
is used to classify alcohols of low molecular
weights. This reaction shows a substitution,
which the chloride replaces a hydroxyl
group. A positive result indicated the
formation of a chloroalkane which is based
on the difference in reactivity of the three
classes of alcohols with hydrogen halides.
The difference in reactivity shows the
different ease of formation of the
carbocations. Tertiary carbons are the most
stable and Primary carbons are the least
stable. Tertiary alcohols react immediately
to with the Lucas Reagent, Secondary
alcohols react within a few minutes while
Primary alcohols do not react with Lucas
Reagent at room temperature. Meaning that
the time taken for a positive effect to be
visible is the measure of the reactivity of the
class of alcohol, and it is also the way of
determining the class of a alcohol the
solution is. Here is the reaction mechanism
in Lucas Test [5].

primary and secondary resulting in the

reduction of the orange chromium Cr6+ ion
to a blue-green Cr3+ ion.
Carboxylic acids and their derivatives are
the most abundant of all organic compounds
in living organisms and in the laboratory.
The most common derivatives of carboxylic
acids are acyl halides, acid anhydrides,
esters, and amides. They contain acyl group
attached to a nucleophilic molecule that
replaced the OH group of a carboxylic
acid. Meaning that Jones test is used to
check for the presence of a primary and
secondary alcohol both of which shows a
positive result of the formation of a green
color. Here is the reaction mechanism in
Chromic Acid Test [6].

Figure 2 : Jones test Rxn Mechanism

Figure 1 : Lucas Test Rxn Mechanism

The Chromic acid test, also known as Jones
Oxidation test, differentiates primary and
secondary alcohols from tertiary alcohols. A
primary alcohol is oxidized to an aldehyde
or to a carboxylic acid, while a secondary
alcohol to a ketone. Tertiary alcohols in the
other hand do not react. The OH-bearing
carbon must have a hydrogen atom attached
since that carbon atom is being oxidized in

The 2,4-Dinitrophenylhydrazone (2,4-DNP)

test serves as to form a derivative. The
aldehydes and ketones react with 2,4-DNP
to form a solid 2,4-DNP Derivative. If the
solid is yellow, this means that the carbonyl
group in the unknown is unconjugated while
a reddish-orange color most likely means
that the carbonyl group is conjugated.
Meaning this test help determine whether
the carbonyl group is alipathic or aromatic.
The reddish-orange color means that the
substance is aromatic and the yellow color

means that the substance is alipathic. Here is

the reaction mechanism in 2,4-DNP [7].

Figure 3 : 2,4-DNP Rxn Mechanism

The Tollens Test, also known as Silver

Mirror Test, is used to distinguish between
an aldehyde and a ketone. This test also has
the same function as the Fehlings Test. This
test uses a reagent known as Tollens
reagent, which is a colorless, basic, aqueous
solution containing solver ions coordinated
to ammonia [Ag(NH3)2+]. Tollens reagent
oxidizes an aldehyde into the corresponding
carboxylic acid. A positive result would
make the reagent metallic silver in color and
shows a mirror-like expression in the test
tube. Ketones are not reactive with the
Tollens reagent, so it would not show a
mirror-like impression in the test tube. Here
is the reaction mechanism in Tollens Test
[9].

The Fehlings test is used to determine the

presence of an aldehyde in the solution and
it doesnt react with ketones. Aldehydes
reduces copper to make the deep blue
solution of copper (II) to muddy green
solution, and then forms a brick-red
precipitate of insoluble cuprous oxide
(Cu2O). Meaning this test is usually used for
reducing sugars but us known to be not
specific with aldehydes. Here is the reaction
mechanism in Fehlings Test [8].

Figure 5 : Tollens Test Rn Mechanism

Figure 4 : Fehlings Test Rxn Mechanism

Lastly, the Iodoform test used to see the

presence of CH3CO in aldehydes and
ketones. The sample is allowed to react with
a mixture of iodine and base. The alpha
hydrogens of the carbonyl group are acidic
and will react with a base to form the anion,
which then reacts with iodine and forms
triiodo compound which then reacts with
another base to form the carboxylic acid salt
plus a iodoform. A positive result would
produce a yellow precipitate, and if there is
no CH3CO group in the sample being tested,
then it will get different result. Here is the
reaction mechanism in Iodoform Test [10].

alcohol was added. The test tube was

covered and shaken vigorously and was
allowed to stand at room temperature. The 2
other test tubes were placed with 2-3 drops
of sec-butyl alcohol and tert-butyl alcohol.
The test tubes were shaken vigorously and
the mixtures were allowed to stand. Note the
time of the formation of the cloudy
suspension or the formation of two layers.
Figure 6 : Iodoform Test Rxn Mechanism
METHODOLOGY
The materials needed for this experiment in
this experiment are the following: Lucas
Reagent, Chromic Acid reagent, 95%
ethanol, Fehlings A and B, Tollens reagent,
5% NaOCl solution, Iodoform test reagent,
2,4-dinitrophenylhydrazine, Pasteur pipette,
test tubes and vials and some beakers. While
the sample compounds needed are the
following: ethanol, n-butyl alcohol, secbutyl alcohol, tert-butyl alcohol, benzyl
alcohol, n-butyraldehyde, benzaldehyde,
acetone, acetophenone, isopropyl alcohol,
acetaldehyde.
A. Solubility of Alcohols in Water
Five test tubes were labelled and ten drops
each of ethanol, n-butyl alcohol, sec-butyl
alcohol, tert-butyl alcohol, and benzyl
alcohol were placed on the test tubes with
the use of a Pasteur pipette. Followed by
addition of 1mL of water and shaking of the
test tube. If cloudiness occurs, continue
adding a quarter of an mL of water with
vigorous shaking, until a homogenous
dispersion results. If no cloudiness results
after the addition of 2mL of water, the
alcohol is water soluble in water.
B. Lucas Test
Three test tubes were prepared and 1mL of
Lucas Reagent was dropped in each of them.
One the first test tube, 2-3 drops of n-butyl

C. Chromic acid test (Jones Oxidation)

Six test tubes were prepared as well as six
samples. The samples are n-butyl alcohol,
isopropyl alcohol, tert-butyl alcohol,
acetaldehyde, benzaldehyde, and acetone.
One drop of each sample was placed on
different test tubes and was dissolved in
1mL of acetone followed by the addition of
5 drops of the reagent dropwise with
continuous shaking. Place the test tubes in
60oC water bath for 5 minutes and note the
color of each solution.
D. 2,4-Dinitrophenylhydrazone Test
Four test tubes were labelled with the
samples to be used. The samples used were
acetone, acetaldehyde, benzaldehyde, and
acetophenone. One drop of each sample was
placed in different test tubes followed by the
addition of 5 drops of 95% ethanol with
continuous shaking. Then add 3 drops of
2,4-DNP. If no yellow or orange-red
precipitation forms, allow the solution to
stand for 15 minutes. Note the results.
E. Fehlings Test
Into each test tube, 1mL 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 into a beaker of boiling water.
Changes were observed that occurred within
10-15 minutes. The samples to be used are
acetaldehyde, acetone, benzaldehyde, and
acetophenone.

F. Tollens Silver Mirror Test

Four test tubes were prepared that contains
1mL of freshly prepared Tollens reagent
and two drops of each sample was placed
into separate test tubes. The samples were
acetaldehyde,
benzaldehyde,
acetone,
acetophenone. Shake the mixture well and
allow it to stand for 10 minutes. If no
reaction has occurred, place the test tubes in
a beaker with warm water for 5 minutes.
Record any observation.
I.
Iodoform Test
2 drops of each sample (acetaldehyde,
acetone, acetophenone, benzaldehyde, and
isopropyl alcohol) were placed in different
test tubes. 10 drops of 10% KI solution were
added. Followed by the addition of 20 drops
of fresh chlorine bleach (5% sodium
hypochlorite) were added slowly to each test
tube with continuous shaking. The formation
of a yellow precipitate was noted.
G. Hydrolysis of Acid Derivatives
Acid Halides and Acid anhydrides
In a test tube, 1mL of water was
placed, and then 10 drops of the sample was
cautiously added dropwise. The resulting
mixture was divided into two portions. 1mL
of 2% AgNO was added to the first test tube
while 1mL of saturated NaHCO3 was added
to the 2nd portion.
Esters
2mL of 25% NaOH solution was
added to 1mL of ethyl acetate. The mouth of
the test tube was covered with a marble then
the test tube was heated in a boiling water
bath for 5 minutes. Afterwards, the mixture
was neutralized with 10% HCl solution. The
test tube was wafted to take note of the odor.

Amides

Here 1 mL of benzamide was treated

with 5mL of NaOH solution and was got
immersed in a boiling water bath. During
heating, a moist red litmus paper was placed
over the test tube and any change in the
litmus paper was noted.
H. Alcoholysis: Schotten-Baumann Rxn
Acetic Acid
In this procedure, 10 drops of acetic
acid, 1mL ethanol, and 5 drops of
concentrated H2SO4 was warmed on a water
bath for 2 minutes and then note the odor of
the ester formed.
Acyl halides and Acid anhydrides
In another test tube, we placed 0.5
mL ethanol, 1mL water, and 0.2mL of acyl
halide or acid anhydride sample. Then add
2mL of 20% NaOH solution. Plug the test
tube using a cork stopper or parafilm and
shake the mixture for several minutes.
Afterwards, note the odor and the layers of
the ester formed.
I. Aminolysis: Anilide Formation
Acyl halides and acid anhydrides
A few drops of either acetyl chloride
or acetic anhydride were added to 0.5mL
aniline. The mixture was then transferred to
a new test tube containing 5mL of water.
Formation of precipitation was then
observed.
J. Hydroxamic Acid Test
Preliminary test was done by mixing
1mL of 95% ethanol and 1M HCl to
separate test tubes that contained drops of
the sample to be used. If upon addition of 1
drop of 5% FeCl3, a color other than yellow
will make the test futile. 2mL of alcoholic
NH2OHHCl and 1mL of 1M KOH was
added to separate test tubes of 2 drops ethyl
acetate and acetamide. Immersion in a
boiling water bath for 2 minutes was done;
afterwards the mixture was allowed to cool.

1mL of 5% FeCl3 was introduced in each

test tube. Then the color of the formed
precipitate was noted.

true for organic compounds that have the

same number of carbon atoms present.

RESULTS AND DISCUSSION

The solubility of alcohols decreases as the

length of the hydrocarbon chain of the
alcohol increases.

For the test of the solubility of alcohols in

water, the turbidity of the solution was
observed and it was the basis if the specific
alcohol is soluble to water. Also, the amount
of water needed to produce homogenous
dispersion was observed. Table 1 shows the
data gathered from the test.

The next test was conducted with the use of

the Lucas Reagent and it differentiated 1,
2 and 3 alcohols. Alkyl chloride formation
was observed and caused turbidity or
cloudiness. Also, the rate of the reaction was
observed. Table 2 shows the result of Lucas
Test.

Alcohol

ethanol
n-butyl alcohol
sec-butyl
alcohol
Tert-butyl
alcohol
Benzyl alcohol

Amount of
water needed
to produce
homogenous
dispersion
1mL
1.50mL
1mL

Solubility
to water

Sample
n-butyl alcohol
Sec-butyl alcohol
Tert-butyl alcohol

Observation
Homogenous solution
Homogenous solution
Formation of 2 layers

Table 2 : Lucas Test results

Soluble
Soluble
Soluble

1mL

Soluble

2mL

Insoluble

Table 1 : Solubility of Alcohols in Water

Table 1 shows the solubility of alcohols to
water and the amount of water that is needed
to produce a homogenous dispersion.Out of
the 5 alcohol samples, only Benzyl Alcohol
is insoluble. The principle behind this is that
like dissolves like. It can be said that the
other alcohols are soluble to water because
they exhibit a polar bond and water is also
known to have a polar bond as well. There
are factors that affect the solubility of
alcohols to water. One of these factors is the
number of carbon atoms present. The lower
the number of carbon atoms present, the
more soluble or miscible a substance is.
Another factor is the branching of carbon
chains. The more branching present, the
more soluble a compound is. This is only

According to the table above, n-butyl

alcohol and sec-butyl alcohol are both
soluble in the Lucas Reagent while tertbutyl alcohol formed a cloudy layer. The
absence of visible reaction at room
temperature and the appearance of the
cloudy layer on the application of heating,
represents that it is a primary alcohol. A
delayed formation of a cloudy layer; around
3-5 minutes tells us that it is a secondary
alcohol. And lastly, if the substance turn
cloudy immediately after the addition of
Lucas reagent and the phases separate means
that it is a tertiary alcohol. Basing on the
table above, tert-butyl alcohol is a tertiary
alcohol due to the immediate clouding of the
alcohol and the separation of layers.
Chromic Acid test (Jones Oxidation) permits
the conversion of primary alcohols to
aldehydes and secondary alcohols to ketones
through oxidation. Table 3 shows the results
of Chromic Acid test.

Sample
n-butyl alcohol
Tert-butyl alcohol
Acetaldehyde
Benzaldehyde
Acetone
Acetophenone
Isopropyl Alcohol

Observation
Blue-green solution
Dirt yellow solution
Blue-green solution
Blue-green solution
Green solution
Dirty yellow solution
Blue-green solution

Table 3 : Chromic Acid test results

In the data gathered, n-butyl alcohol,
acetaldehyde, benzaldehyde, and isopropyl
alcohol gave a positive result of a blue-green
solution while tert-butyl alcohol and
acetophenone both gave a dirty yellow
solution while acetone gave a green
solution. Chromic test or Jones Oxidation
involved reduction-oxidation or redox
reaction. 1 and 2 alcohols and aldehydes
went through oxidation and chromium
underwent reduction from Cr6+ to Cr3+. The
reduction of the chromium ion which is
colored orange resulted to a blue-green
solution telling us that the 1 and 2
alcohols were oxidized. Ketones are the
result of the reaction of 2 alcohols with
chromic acid, which do not oxidize further.
Tertiary alcohols are nonreactive and
aldehydes got oxidized to carboxylic acids.
The
next
test,
which
is
2,4Dinitrophenylhydrazne test / 2,4-DNP is the
test used for distinguishing the presence of
an aldehyde or ketone in a compound. Table
4 shows the data gathered from the test.
Sample
Acetaldehyde
Benzaldehyde
Acetone
Acetophenone

Observation
Yellow-orange ppt.
Yellow-orange ppt.
Canary yellow ppt.
Orange-red ppt.

Table 4 : 2,4-DNP test results

In the test, acetophenone was the only

sample to give an orange-red ppt. while the
other samples gave a yellow-orange and
canary yellow precipitates. The orange-red
precipitate represents the presence of a
conjugated carbonyl compound and the
appearance of the yellow precipitate
indicates the presence of unconjugated
carbonyl compound.
Fehlings Test is another differentiating test
for aldehydes and ketones. In this test
aldehydes reacted in this test while ketones
did not produce any reaction.
Sample
Acetaldehyde
Benzaldehyde
Acetone
Acetophenone

Observation
Brick-red ppt.
Brick-red ppt.
Dark blue solution
Royal blue solution

Table 5 : Fehlings test results

As shown on the table below above,
acetaldehyde and benzaldehyde both
produced a brick-red precipitate indicating
that they are both aldehydes. Meanwhile
acetone and acetophenone did not produce a
reaction what so ever. Fehlings test
involved reduction-oxidation or redox
reaction. Aldehydes got oxidized while
ketones did not undergo oxidation.
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 on Tollens Silver Mirror
test.
Sample
Acetaldehyde
Benzaldehyde
Acetone
Acetophenone

Observation
Silver mirror
Gray soln w/ globules
Colorless soln
Turbid gray soln

Table 7 : Tollens Silver Mirror test results

On the table above, only acetaldehyde
formed a silver mirror. Benzaldehyde, even

though it is an aldehyde did not form any

silver mirror. Acetone formed a colorless
solution and acetophenone produced a turbid
gray solution. This test is similar to
Fehlings test making it undergo redox
reaction as well. Aldehydes got oxidized
while ketones did not.
Iodoform test was used to detect the
presence of an aldehyde or a ketone in a
substance where methyl groups are attached
directly to the carbonyl carbon.
Sample
Acetaldehyde
n-butylraldehyde
Benzaldehyde
Acetone
Acetophenone
Isopropyl Alcohol

Observation
Yellow ppt.
Yellow solution
Red ppt. w/ globules
Yellow ppt.
Yellow ppt.
Yellow crystal ppt.

Table 7 : Iodoform test results

As shown in the table above, acetaldehyde,
acetone, and acetophenone produced a
yellow precipitate. Benzaldehyde resulted
produced a red precipitate with globules.
Isopropyl alcohol resulted in a yellow
crystalline precipitate and n-butyraldehyde
resulted in a yellow solution. The
appearance of a yellow precipitate or a
yellow crystal indicates a positive result. It
also tells us that the carbonyl group of the
sample has a methyl group attached directly
to it.
REFERENCES
From books
1. Brown, W., Poon, T. (2011).
Introduction to organic chemistry
international student version (5th
edition). NJ, USA: John Wiley &
Sons, Inc.
2. Zumdahl, S., Zumdahl, S. (2012).
Chemistry: An Atoms First Approach
(International edition). USA:
Brooks/Cole, Cengage Learning.

From the Internet

3. Clark, J. (2003). An Introduction to
Alcohols. Retrieved from
http://www.chemguide.co.uk/organic
props/alcohols/background.html
4. Clark, J. (2003). Aldehydes and
Ketones. Retrieved from
http://www.chemguide.co.uk/organic
props/carbonyls/background.html
5. Lucas Reagent (n.d.) Retrieved from
http://www.chemistrylearner.com/luc
as-reagent.html
6. Test for Aldehydes and Ketones (n.d.)
Retrieved from
http://academics.wellesley.edu/Chem
istry/chem211lab/Orgo_Lab_Manual
/Appendix/ClassificationTests/aldehy
de_ketone.html
7. 2,4-Dinitrophenylhydrazine test
(n.d.) Retrieved from
http://www.harpercollege.edu/tmps/chm/100/dgodambe/thedisk/qual/
dnp.html
8. Fehlings Test (n.d.) Retrieved from
https://fenix.tecnico.ulisboa.pt/downl
oadFile/3779571247498/Testes
%20de%20a%C3%A7ucaresalunos.pdf
9. Tollens Test (n.d.) Retrieved from
http://chemwiki.ucdavis.edu/?
title=Organic_Chemistry/Aldehydes
_and_Ketones/Reactivity_of_Aldehy
des_%26_Ketones/Tollens
%E2%80%99_Test
10. Clark, J. (2004). Iodoform Reaction
with Aldehydes and Ketones.
Retrieved from
http://www.chemguide.co.uk/organic
props/carbonyls/iodoform.html