MODERN INSTITUTE OF

PHARMACEUTICAL SCIENCES
GRAM ALWASA, BEHIND REWATI RANGE, SANWER ROAD, INDORE, 453111
Website: www.moderninstitutes.in Email: mipscollege@gmail.com

Academic Year: 2019-2020

Subject: Pharmaceutical Organic Chemistry
Topic: Qualitative test for different functional group

Name of Faculty: Ms. Prashansa Nema

Assistant Professor

MODERN INSTITUTE OF PHARMACEUTICAL SCIENCES

Modern Modern Nandani Medical

Incubator Laboratories Laboratories
 Index
Index
S. No. Particular Page No.
Qualitative test for the following
functional group :-
1. Alcohols 4-5
2. Qualitative Test for Alcohols 6-21
3. Carboxylic acid 22-23
4. Qualitative Test for Carboxylic acid. 24-33
5. Amide 34-35
6 Qualitative test for Amide 36-42
7. Ester 42-43
8. Qualitative test for Ester 44-47

MODERN INSTITUTE OF PHARMACEUTICAL SCIENCES

Modern Modern 2
Nandani Medical
Incubator Laboratories Laboratories
 Qualitative test for the following
functional group

1. Alcohol
2. Carboxylic Acid
3. Amide
4. Ester

3
 1. ALCOHOLS
Alcohol, Alcohol, sometimes referred to by the chemical name
ethanol, is a psychoactive drug that is the active ingredient in
drinks such as beer, wine, and distilled spirits (hard liquor).
It is one of the oldest and most common recreational
substance, causing the characteristic effects of alcohol
intoxication ("drunkenness").

Ethanol
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• Among other effects, alcohol produces a mood lift and
euphoria, decreased anxiety, increased sociability, sedation,
impairment of cognitive, memory, motor, and sensory function,
and generalized depression of central nervous system function.

Ethanol is only one of several types of alcohol, but it is the only

type of alcohol that is found in alcoholic beverages or
commonly used for recreational purposes;
other alcohols such as methanol and isopropyl alcohol.
The amount of ethanol in the body is typically quantified by
blood alcohol content (BAC); weight of ethanol per unit
volume of blood.

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 QUALITATIVE TEST FOR ALCOHOLS
• In this experiment you are going to do a series of tests in
order to determine whether or not an alcohol is a primary
(1°), secondary (2°) or tertiary (3°) alcohol.
• The tests can also determine whether or not there is a
• secondary methyl alcohol functionality in the molecule.
• You will do four chemical tests:
(1) Chromic Acid Test(or Jones Oxidation),
(2) Ritter Test using potassium permanganate
(3) the Lucas Test using ZnCl2 and HCl,
(4) the Iodoform Test.

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• There will be perform each of the first three tests three times,
once with a primary, once with a secondary and once with a
tertiary alcohol.

• The fourth test will do two times, once with a secondary

methyl containing alcohol and once with a not secondary
methyl alcohol.

• The tests on the three known alcohols can be done at the

same time.

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 Test 1 : Chromic acid
oxidation
• This test 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.
• Recall, that a carbon is oxidized when it loses a hydrogen or
hydrogens or gains a more electronegative atom.
• 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 6.1.

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9
 Procedure:
• Set-up three small test tubes in your test tube rack.
• The tubes do not need to be dry.
• Label the first test tube as a primary alcohol, the next as a
secondary alcohol and the third as a tertiary alcohol. Write
down which alcohols you are going to be using.

• Add 2 mL of acetone to each test tube and then add 3-

4 drops of your test alcohol.
Be sure that the drops fall into the acetone and do not remain
on the sides of the tube.
Add 2 drops of the Chromic Acid Test Reagent (also called the
Bordwell-Wellman Reagent).

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• Shake vigorously using a small, tight fitting cork.
• You should see a color change to a blue or blue-green or
similarly colored precipitate within a few seconds to indicate a
positive test.
• Record the result.

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 Test 2: Ritter Test

• This test is similar to the Chromic Acid Oxidation and provides

the same information.
• It is the oxidation of primary and secondary alcohols to
carboxylic acids and ketones using potassium permanganate
(KMnO4).
• Again, tertiary alcohols cannot be oxidized by this reagent
because there is no hydrogen to be lost from the carbon that
bears the OH group.
• In the Ritter Test the Mn7+ of KMnO4 (bright purple) is
reduced to Mn4+.
• The Mn4+ is brownish in color.
The reactions involved are as follows.
12
+KMnO4
& AcOH

+KMnO4 &
AcOH

13
 Procedure:
• As for the Chromic Acid Oxidation, set-up three small, labeled
tests tubes.
• Add 2 mL acetic acid to each tube. Add 3-4 drops of the test
alcohol to each tube and then add ONE drop of saturated
KMnO4 solution to each test tube.
• Shake vigorously to mix, using a small cork as before.
• For the 1°and 2° alcohols you should see a brownish color
develop as the purple KMnO4 color disappears.
• Do not add too much KMnO4. If you add an excess of this
reagent, the purple color will persist even though you have a
primary or secondary alcohol.

• With the tertiary alcohol you should see no color change since
the purple color remains.
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 Test 3: Lucas Test

• This test distinguishes tertiary, secondary and primary

alcohols from each other.
• It uses zinc chloride as the reagent in concentrated
hydrochloric acid (Lucas Reagent).
• It is based on the rate of formation of insoluble alkyl
chloride.
• An emulsion is formed.
• This test is reliable only for alcohols that are fairly soluble
in water.

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• Tertiary Alcohols : React immediately to form an emulsion of
the alkyl halide and water (cloudysolution).

• Secondary Alcohols : React in 5-10 minutes.

• Heating in warm water and shaking is sometimes necessary
with water-insoluble alcohols.

• Primary alcohols :These generally take more than one hour to

react.
• The rate of reaction of the alcohols is determined by the ease
of formation of the carbocation intermediate.
• Tertiary alcohols react quickly because they form a relatively
stable tertiary carbocation and
• secondary alcohols react more slowly because the secondary
carbocation is less stabilized than the tertiary carbocation.
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17
 Procedure:

• Set-up three small test tubes as above, labeling each of

them.
• Add 0.5 mL of the test alcohol to each test tube.
• To the first test tube, add 3 mL of the Lucas reagent, Shake
vigorously using a small cork to stopper the test tube.
• Wait at least 10 minutes. If you have a water insoluble
alcohol (you see two layers in your test tube) then repeat
the experiment using a hot water bath set at 60 °C,
immersing the test tube in the water bath immediately
after shaking and waiting the 10 minutes with occasional
shaking.
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 Test 4: Iodoform Test

• This test is slightly different from the previous three tests.

• This test does not distinguish 1°, 2°, 3° alcohol but is specific
for only one class of alcohol.
• This is the secondary methyl alcohol.
• If the alcohol contains a methyl group attached to a carbon
that also has a hydrogen and an OH group then it will give a
positive iodoform test.
The formation of a yellow precipitate indicates a positive test.

19
 R = H, CH3, CH2CH3 etc.
R

20
Procedure
To each test tube, add 4-5 drops of test alcohol, 2 mL H2O, 2 mL
10% sodium hydroxide solution.
If your alcohol does not fully dissolve, add 1 mL of ethanol.
Add a 10% solution of iodine (I2) in potassium iodide (KI)
dropwise. Keep adding the dark iodine solution with stirring until
you see a faint yellow color persisting in the test tube.
Note the reaction that is occurring.
As you add the I2 to the NaOH solution, you are forming your
sodium hypoiodite reagent.

21
 1. Carboxylic acid
• A carboxylic acid is an organic compound that contains a
carboxyl group (C(=O)OH).
• The general formula of a carboxylic acid is R–COOH, with R
referring to the alkyl group.
• Carboxylic acids occur widely. Important examples include the
amino acids and acetic acid.
• Deprotonation of a carboxyl group gives a carboxylate anion.

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3 D structure of carboxylic acid
• Carboxylic acids are versatile organic compounds.
• It has excellent physical and chemical properties.
• The carboxylic acid chemical structure contains a carbonyl
functional group and hydroxyl group.
R-COOH in which –OH is hydroxyl group
C=O is carbonyl group
R= alkyl group.
• It interact easily with polar compounds and contributes to
many important chemical reactions.
• Carboxylic acids have a tendency to donate protons and act as
acids.
• It is this property which is helpful in the identification of a
-COOH group.

23
QUALITATIVE TEST FOR CARBOXYLIC
ACID
Any of the following test can be carried out to detect the
carboxylic functional group.

1. Litmus test
2. Sodium bicarbonate test (or) Sodium hydrogencarbonate test
3. Ester test
4. Fluorescein test

24
 Test 1 : Litmus Test

• The carboxylic acid turns blue litmus red.

• Because the hydroxyl group present in -COOH is far acidic
than in alcohol.

The chemical reaction is given below.

R-COOH + H2O → R-COO– + H3O+

If the colour of the blue litmus paper changes to red then

carboxylic group is present.

25
 Procedure
• Add a drop of given organic compound on blue litmus paper.

• Observe the colour change in blue litmus paper.

• If the colour of blue litmus changes to red the presence of

carboxylic acid.

Blue litmus solution is also used in the place of blue litmus paper.

26
 Test 2 : Sodium Bicarbonate Test

• When carboxylic acid reacts with sodium bicarbonate

solution carbon dioxide is evolved with a brisk
effervescence along with sodium acetate is formed.

• The chemical reaction is given below.

RCOOH + NaHCO3 → RCOONa + H2O + CO2↑

acetic acid sod.bicarbonate Sod. acetate (brisk effervescence)

This test is used to distinguish between carboxylic acid

from phenol. Phenol does not answer to this test.
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 Procedure
• Prepare a saturated solution of sodium bicarbonate by
dissolving sodium bicarbonate in 1ml of water.

• Add the given organic compound on the saturated solution

of sodium bicarbonate solution.

• Shake the solution well.

• If there is an evolution of brisk effervescence then it

indicates the presence of carboxylic acid.
Use acid free alcohol for the test.
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 Test 3 : Ester Test
• Carboxylic acid reacts with alcohol in the presence of
concentrated sulfuric acid forms a pleasant smelling
ester.
• This reaction is known as esterification.

The chemical reaction is given below.

RCOOH + R-OH + H2SO4 → RCOO-R (Ester) + H2O

Formation of a sweet smelling compound indicates the

presence of carboxylic group in the given organic
compound.
29
 Procedure
• Mix the given compound with ethyl alcohol and concentrated
sulfuric acid.

• Heat the mixture in a dry test tube in a water bath.

• Pour the reaction mixture into a beaker carefully containing

water.

• Neutralise the excess sulfuric acid.

• If a sweet smelling substance is sensed then it indicates the

presence of acid.

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 Test 4 : Fluorescein Test
This test is given by dicarboxylic acid.
• Dicarboxylic acid on heating gives acid anhydride.
• When this anhydride is treated with resorcinol in the presence
of concentrated sulfuric acid a fluorescent dye is formed and so
this reaction is called fluorescein test.
The chemical reaction is given below.

This test should be performed only if the compound gives positive

results in litmus test and sodium bicarbonate test.
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 Procedure
• Take the given organic compound to be tested in a test
tube.
• Mix with 100mg of resorcinol and 0.5ml of concentrated
sulfuric acid.
• Heat the test tube gently on a bunsen burner.
• Pour the mixture into a beaker containing dilute sodium
hydroxide solution.
• Appearance of green colour fluorescent solution
indicates the presence of carboxylic acid group.
The resultant solution should be alkaline.

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Litmus Test Carboxylic group turns blue litmus
red.

Sodium bicarbonate test Brisk effervescence indicates the

presence of carboxylic acid

Formation of a sweet smelling

Ester test compound indicates the presence
of carboxylic group.

Appearance of green colour

Fluorescein test fluorescent solution indicates the
presence of acid group.
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 3. Amide
An amide, also known as an organic amide or a carboxamide, is
a compound with the general formula RC(=O)NR′R″, where R,
R′, and R″ represent organic groups or hydrogen atoms.
It can be viewed as a derivative of a carboxylic acid RC(=O)OH
with the hydroxyl group –OH replaced by an amine group –NR
′R″; or, equivalently, an acyl (alkanoyl) group RC(=O)– joined
to an amine group.

3-D structure of
Structure of amide amide
34
• Common examples of amides : are acetamide H3C–CONH2,
benzamide C6H5–CONH2, and dimethylformamide HCON(–
CH3)2.
• Amides are qualified as primary, secondary, and tertiary
according to whether the amine subgroup has the form –NH2,
–NHR, or –NRR′, where R and R′ are groups other than
hydrogen.
• The core –C(=O)N= of amides is called the amide group
(specifically, carboxamide group).
• Amides include many other important biological compounds,
as well as many drugs like paracetamol, penicillinand LSD.
• Low molecular weight amides, such as dimethylformamide,
are common solvents.

35
 Qualitative test for amide
Any of the following test can be carried out to detect the amide
functional group.

1. Sodium Hydroxide test

2. Hydroxamic acid test
3. Nitrous acid test
4. Hydrogen Peroxide test

36
 Test 1: Sodium Hydroxide Test
Amides are compound that contain the amide group
(-CONH2) are investigated using the sodium hydroxide solution
test.
Amides react with Sodium Hydroxide producing ammonia gas, which
reacts with hydrochloric acid producing white fumes of ammonium
chloride gas.
Amides are decomposed by NaOH to evolve ammonia. The gas can
be tested by a moist red litmus paper which is then turned blue.

RCONH2 + NaOH ==> RCOONa + NH3

amide sod. Hydroxide Sod. acetate ammonia gas

NH3 + HCl  NH4Cl

( ammonium Chloride gas white fumes)
37
 Test 2:Hydrolysis under acidic
condition
• Taking ethanamide as a typical amide.
• If ethanamide is heated with a dillute acid (such as dillute
hydrochloric acid), ethanoic acid is formed together with
ammonium ions.

CH3CONH2 + H2O + HCl  CH3COOH + NH4+ Cl-

(Amide) (Acetic acid)
In this reaction , HCl is acting as both catalyst and secondly
reacting with those ethanoate ions to make ethanoic acid.

38
Alkaline hydrolysis of aromatic amides to aromatic acid:

The soluble sodium salt of aromatic acid formed from

aromatic amides upon hydrolysis is regenerated as white
precipitate in acidic medium.
•  

39
 Biuret Reaction for aliphatic diamide:

When aliphatic diamide is heated at a temperature above its

melting point, ammonia is evolved and crystalline biuret is
formed. This biuret in alkaline medium gives a violet colour
with a drop of copper sulphate solution.

Biuret Reaction
40
 Test 3: Hydrogen Peroxide Test
• Hydrogen peroxide reacts with aromatic primary amides to
form the hydroxamic acid, which then reacts with ferric
chloride to form ferric hydroxamate complex having a violet
colour.
•  

41
Summary of all the test for amide

42
 4. Ester
• an ester is a chemical compound derived from an acid
(organic or inorganic) in which at least one –OH (hydroxyl)
group is replaced by an –O–alkyl (alkoxy)group.

• Usually, esters are derived from a carboxylic acid and an

alcohol.

3-D struture of Ester

43
• Glycerides, which are fatty acid esters of glycerol, are
important esters in biology, being one of the main classes of
lipids, and making up the bulk of animal fats and vegetable
oils.
• Esters with low molecular weight are commonly used as
fragrances and found in essential oils and pheromones.
• Phosphoesters form the backbone of DNA molecules. Nitrate
esters such as nitroglycerin, are known for their explosive
properties, while polyesters are important plastics, with
monomers linked by ester moieties.

44
 Qualitative test for ester
1. Phenolphthalein test
2. Hydroxamic acid test

• Since esters are usually colourless liquids,

• there is no simple test to detect them, apart from their
pleasant 'odour'.
• Some tests involve reacting the ester with saturated ethanolic
hydroxylamine hydrochloride + 20% methanolic KOH and
gently heated until boiling.
• Then mixture acidified with 1M HCl (aq) and FeCl3 (aq) added
dropwise.

45
 Test 1 : Hydroxamic acid test
• In this test, ester is converted to a hydroxamic acid
(HOHN-C=O), which will give positive ferrric chloride test.
If a color forms and then fades, add additional drops of 5%
ferric chloride until the color persist.

Ester

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47
 Reference
• www.thestudentroom.co.uk/showthread.php?t=4197
86
• myweb.liu.edu/~swatson/downloads/files/Experiment
_6.pdf.
• www.quora.com/How-are-carboxylic-acids-identified
• https://www.chemguide.co.uk/organicprops/amides
/hydrolysis.html
• amrita.olabs.edu.in/?
sub=73&brch=8&sim=141&cnt=1
• https://vlab.amrita.edu/?sub=2&brch=191&sim=345
&cnt=1
48
Thank You

49