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,
Fehling’s 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 blue-
green 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. Fehling’s 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 Fehling’s
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 number of carbon substituents bonded to the

hydroxyl-bearing carbon (McMurry, 2010). The
An alcohol is a compound that has a most important physical property of alcohols is
hydroxyl group bonded to a saturated, spᶟ- the polarity of their –OH groups. Because of the
hybridized carbon atom, R-OH (Figure 1). large difference in electronegativity between
Alcohols are classified as primary (1˚), oxygen and carbon and between oxygen and
secondary (2˚), or tertiary (3˚), depending on the hydrogen, both the C-O and O-H bonds of an
alcohol are polar covalent, and alcohols are carbocation, which in turn reacts with the
polar molecules (Brown & Poon, 2011). chloride ion (nucleophile) to generate an alkyl
Alcohols usually have much higher boiling halide product. Figure 4 shows the general
points than might be expected from their molar mechanism for this SN1 reaction.
masses. This difference can be understood if we
consider the types of intermolecular attractions
that occur in these liquids (Zumdahl & Zumdahl,
2012).

Figure 1. Structure of Alcohol

The functional group of an aldehyde is a

carbonyl group bonded to a hydrogen atom Figure 4. General Mechanism of Lucas test
(Figure 2). The functional group of a ketone is a
carbonyl group bonded to two carbon atoms The Chromic acid test (Jones oxidation)
(Figure 3). Because of the polarity of the distinguishes primary and secondary alcohols
carbonyl group, aldehydes and ketones are polar from tertiary. Chromic acid will oxidize a
compounds and interact in the liquid state by primary alcohol first to an aldehyde and then to
dipole-dipole interactions. As a result, aldehydes a carboxylic acid and it will oxidize a secondary
and ketones have higher boiling points than alcohol to a ketone. Tertiary alcohols do not
those of nonpolar compounds with comparable react. The OH-bearing carbon must have a
molecular weight (Brown & Poon, 2011). 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 Figure 5. General Mechanism of Jones

Oxidation
The Lucas reagent is an aqueous
solution of strong acid (HCl) and zinc chloride The 2,4-Dinitrophenylhydrazone test
(ZnCl₂). The alcohol starting material must be serves as a derivative formation. Both aldehydes
sufficiently soluble in aqueous environments for and ketones react with 2,4-
the reaction to take place. The reaction that dinitrophenylhydrazine to form a solid 2,4-
occurs in the Lucas test is an SN1 nucleophilic dinitrophenylhydrazone (DNP) derivative. The
substitution. The acid catalyst activates the OH color of this derivative can also provide useful
group of the alcohol by protonating the oxygen structural information. If the solid is yellow, this
atom. The C-OH₂+ bond breaks to generate the most often means that the carbonyl group in the
unknown is unconjugated. A reddish-orange oxidized, whereas ketones are not. Tollens’ test
color most likely means that the carbonyl group uses a reagent known as Tollens’ reagent, which
is conjugated. In a few cases, compounds in is a colorless, basic, aqueous solution containing
which the carbonyl group is not conjugated silver ions coordinated to ammonia [Ag(NH3)2+].
produce orange precipitates. Simply having a Tollens’ reagent oxidizes an aldehyde into the
double bon or phenyl group somewhere in an corresponding carboxylic acid. The reaction is
aldehyde or ketone does not necessarily mean accompanied by the reduction of silver ions in
that the carbonyl group is conjugated. The Tollens’ reagent into metallic silver, which, if
double bond must be separated from the the test is carried out in a clean glass test tube,
carbonyl by one single bond only. If the double forms a mirror on the test tube. Ketones are not
bond is further away, it is isolated from the oxidized by the Tollens’ reagent, so the
carbonyl and not conjugated with the carbonyl. treatment of a ketone with Tollens’ reagent in a
Figure 6 illustrates the reaction occurring in the glass test tube does not result in a silver mirror.
2,4-DNP/2,4-DNPH test. 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 2,4- The Iodoform test indicates the presence
Dinitrophenylhydrazone test of an aldehyde or ketone in which one of the
groups directly attached to the carbonyl carbon
In Fehling’s test, the presence of is a methyl group. Such a ketone is called a
aldehydes and not ketones is detected by methyl ketone. In the Iodoform test, the sample
reduction of the deep blue solution of copper (II) is allowed to react with a mixture of iodine and
to muddy green solution, and then form a brick- base. Hydrogens alpha to a carbony group are
red precipitate of insoluble cuprous oxide acidic and will react with base to form the anion,
(Cu2O). This test is commonly used for reducing which then reacts with iodine in this way to form
sugars but is known to be not specific for the triiodo compound, which the reacts with
aldehydes. Figure 7 shows the general more base to form the carboxylic acid salt plus
mechanism undergone in the Fehling’s test. 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 7. General Mechanism of Fehling’s Figure 9 illustrates the reaction involved in the
test Iodoform 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
 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,
Figure 9. Genral Mechanism of Iodoform test respectively, shook vigorously and the mixtures
were allowed to stand. The rate of formation of
One of the objectives of this experiment cloudy suspension or the formation of two layers
is to distinguish whether a compound is was observed.
hydroxyl- or carbonyl-containing. Other
objectives include differentiating the three types 3. Chromic Acid test (Jones Oxidation)
of alcohols, differentiating aldehydes from
Using the samples n-butyl alcohol, sec-butyl
ketones, and explaining the mechanisms
alcohol, tert-butyl alcohol, n-butyraldehyde,
involved in the differentiating tests.
benzaldehyde, acetone, and acetophenone, one
Materials and Methods drop of each liquid or small amount of the solid
sample was dissolved in 1 mL of acetone in
A. Materials different test tubes. 2 drops of 10% aqueous
K2Cr2O7 solution and 5 drops of 6 M H2SO4
In this experiment, the materials needed are were added in each of the test tubes.
Lucas reagent, chromic acid reagent, 95%
ethanol, Fehling’s A and B, Tollens’ reagent, 4. 2,4-Dinitrophenylhydrazone (2,4-
5% NaOCl solution, iodoform test reagent, 2,4- DNP/2,4-DNPH) test
dinitrophenylhydrazine, Pasteur pipette, test
tubes, beaker, and the sample compounds One drop of a liquid sample (acetone,
ethanol, n-butyl alcohol, sec-butyl alcohol, tert- acetaldehyde, n-butylraldehyde, benzaldehyde
butyl alcohol, benzyl alcohol, n-butyraldehyde, and acetophenone) was placed in a test tube and
benzaldehyde, acetone, acetophenone, isopropyl 5 drops of 95% ethanol were added. After
alcohol and acetaldehyde. shaking well, 3 drops of 2,4-
dinitrophenylhydrazine were added. The
B. Methods solution was allowed to stand for at least 15
minutes when no yellow or orange-red
1. Solubility of Alcohols in Water precipitate forms.

In the five test tubes, 10 drops each of 5. Fehling’s test

ethanol, n-butyl alcohol, sec-butyl alcohol, tert-
butyl alcohol and benzyl alcohol were placed Into each test tube, 1 mL of freshly prepared
using a Pasteur pipette. 1 mL of water was Fehling’s reagent (made by mixing equal
added dropwise in each of the test tubes amounts of Fehling’s A and Fehling’s B) was
containing alcohol while shaking the mixture placed. 3 drops of the sample to be tested were
after each addition. When cloudiness resulted, added and the test tubes were placed in a beaker
0.25 mL of water was added at a time, with of boiling water. Changes were observed that
vigorous shaking, until a homogenous dispersion occurred within 10-15 minutes. The test was
appeared. The total volume of water added was performed on acetaldehyde, n-butyraldehyde,
noted. When no cloudiness resulted after the acetone, benzaldehyde and acetophenone.
addition of 2.0 mL water, alcohol was noted
6. Tollens’ Silver Mirror test
down to be soluble in water.

2. Lucas test
 Five test tubes with 1 mL each of freshly butyl alcohol, sec-butyl alcohol and tert-butyl
prepared Tollens’ reagent were prepared. In each alcohol were all soluble in water. This follows
of the test tubes, 2 drops of each of the samples the principle “like dissolves like” and therefore,
(acetaldehyde, benzaldehyde, acetone, n- it can be said that the alcohols that were soluble
butyraldehyde and acetophenone) were added in water are polar compounds since water is
separately. The mixtures were shaken and polar. Of the alcohols that were soluble in water,
allowed to stand for 10 minutes. When no ethanol, sec-butyl alcohol and tert-butyl alcohol
reaction occurred, the test tubes were placed in a all required only 1 mL of water to be added in
beaker with warm water for 5 minutes. order to be considered soluble. This indicates
Observations were recorded. that there are certain factors affecting solubility.
One of these is the presence of number of carbon
7. Iodoform test atoms. The lower the number of carbon atoms
present, the more soluble or more miscible a
2 drops of each sample (acetaldehyde, acetone, substance is. Branching of carbon chains also
acetophenone, benzaldehyde and isopropyl affect solubility. The more branching present,
alcohol) were placed in different test tubes. 20 the more soluble a compound is. This is only
drops of 10% KI solution were added. While true for organic compounds that have the same
shaking, 20 drops of fresh chlorine bleach (5% number of carbon atoms present.
sodium hypochlorite) were added slowly to each
tube and mixed. The formation of a yellow The Lucas test differentiated 1˚, 2˚ and
precipitate was noted. 3˚ alcohols. Alkyl chloride formation was
observed and caused turbidity or cloudiness. The
Results and Discussion rate of reaction was also observed. Table 2
For the test for solubility of alcohols in presents the results of the Lucas test.
water, turbidity of the solution was to be
Table 2. Lucas test
observed. Cloudiness of the solution indicated
insolubility of that specific alcohol to water. The Sample Reaction observed
amount of water needed to produce homogenous
n-butyl alcohol colorless
dispersion was also observed. Table 1 shows the
sec-butyl alcohol slightly turbid
data gathered from the test.
tert-butyl alcohol turbid
Table 1. Solubility of Alcohols in Water

Alcohol Amount of Solubility to According to the table above, n-butyl

water (in water alcohol was soluble in Lucas reagent while sec-
mL) needed butyl alcohol and tert-butyl alcohol were
to produce a observed to have a formation of cloudy layer.
homogenous Tert-butyl alcohol took the shortest time to form
dispersion the layer while sec-butyl alcohol took the
ethanol 1 mL soluble longest time. The reaction mechanism involved
n-butyl 1.50 mL soluble in the Lucas test is based on SN1 reaction, which
alcohol depends on the formation of stable carbocations.
Reactivity of alcohols in SN1 reaction is 3˚ > 2˚
sec-butyl 1 mL soluble
> 1˚. 3˚ alcohols formed the second layer in less
alcohol
than a minute. 2˚ alcohols required 5-10 minutes
tert-butyl 1 mL soluble while 1˚ alcohols were usually unreactive. The
alcohol presence of ZnCl2, a good Lewis acid, made the
benzyl alcohol 2 mL insoluble reaction mixture even more acidic; thus, it
enhanced the formation of carbocations.
As indicated in the table, only benzyl
alcohol was insoluble in water, while ethanol, n-
 The Chromic Acid test (Jones acetone yellow ppt
Oxidation) tested for oxidizables or any acetophenone red-orange ppt
compounds that possess reducing property (has
an alpha acidic hydrogen). Table 3 shows the
results gathered from the said test. As indicated in the table, only
acetophenone gave a result of red-orange
Table 3. Chromic Acid test (Jones Oxidation) precipitate while the rest of the samples gave a
result of yellow precipitate. A result of red-
Sample Reaction observed orange precipitate indicated the presence of
n-butyl alcohol blue-green solution conjugated carbonyl compounds while a result
sec-butyl alcohol blue-green solution of yellow precipitate indicateed the presence of
tert-butyl alcohol orange solution unconjugated carbonyl compounds. The reaction
n-butyraldehyde blue-green solution of 2,4-DNPH with aldehydes and ketones in an
benzaldehyde blue-green solution acidic solution is a dependable and sensitive test.
acetone orange solution Its reaction mechanism involved condensation or
acetophenone orange solution nucleophilic addition of NH2 to C=O and
elimination of H2O. Some high molecular
weight ketones may fail to react or may yield
According to Table 3, n-butyl alcohol, oils. Most aromatic aldehydes and ketones
sec-butyl alcohol, n-butyraldehyde and produce red dinitrophenylhydrazone while many
bezaldehyde gave a positive result of blue-green nonaromatic aldehydes and ketones produced
solution while tert-butyl alcohol, acetone and yellow products.
acetophenone gave a result of orange solution.
Chromic Acid test / Jones Oxidation involved Fehling’s test was another
reduction-oxidation or redox reaction. 1˚ and 2˚ differentiating test for aldehydes and ketones. In
alcohols and aldehydes underwent oxidation and this test, aldehydes gave a positive result of
chromium underwent reduction from Cr6+ to brick-red precipitate while ketones did not
Cr3+. 1˚ and 2˚ alcohols and aldehydes reduced produce any reaction. Table 5 presents the
the orange-red chromic acid/sulphuric acid results of the test.
reagent to an opaque green or blue suspension of
Table 5. Fehling’s test
Cr (III) salts in 2-5 seconds. 1˚ alcohols reacted
with chromic acid to yield aldehydes, which are Sample Reaction observed
further oxidized to carboxylic acids. 2˚ alcohols
acetaldehyde brick-red ppt
reacted with chromic acid to yield ketones,
n-butyraldehyde brick-red ppt
which do not oxidize further. 3˚ alcohols were
usually unreactive and aldehydes were oxidized benzaldehyde brick-red ppt
to carboxylic acids. acetone blue solution
acetophenone blue solution
The 2,4-Dinitrophenylhydrazone (2,4-
DNP/2,4-DNPH) test detected the presence of
carbonyl groups and tests positive for aldehydes As shown in the given table, only
and ketones. Table 4 shows the results from the acetone and acetophenone did not react to form
test. a precipitate while the rest gave a positive result
of brick-red precipitate. Fehling’s test involved
Table 4. 2,4-Ditrophenylhydrazone (2,4- reduction-oxidation or redox reaction.
DNP/2,4-DNPH) test Aldehydes were oxidized to carboxylic acids
while ketones did not undergo oxidation. In
Sample Reaction observed here, copper was reduced from Cu2+ to Cu1+.
acetaldehyde yellow ppt
n-butyraldehyde yellow ppt Tollens’ Silver Mirror test differentiated
benzaldehyde yellow ppt aldehydes from ketones wherein aldehydes were
expected to be oxidized while ketones did not a result of red precipitate with globules while
undergo any oxidation. Table 6 shows the results isopropyl alcohol gave a result of yellow crystal
from the said test. precipitates. No reaction was observed from n-
butyraldehyde. In this test, yellow crystals or
Table 6. Tollens’ Silver Mirror test precipitate gave a positive result. An alkaline
solution of sodium hypoiodite, formed from
Sample Result observed sodium hydroxide and iodine, converted
acetaldehyde silver mirror acetaldehyde and aliphatic methyl ketones into
n-butyraldehyde flesh solution iodoform (haloform reaction). Since the reagent
benzaldehyde light yellow solution was also an oxidizing agent, alcohols which are
with globules readily oxidized to acetaldehydes or methyl
acetone dark-gray solution ketones also gave a positive reaction. The
acetophenone turbid gray solution mechanism of iodoform synthesis occurred
through a series of enolate anions, which are
iodinated; hydroxide displaced the Cl3- anion
According to Table 6, only acetaldehyde through an addition/elimination pathway.
formed a silver mirror. The samples n-
butyraldehyde and benzaldehyde, although they References
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