Analysis of Common Functional Groups

Experimental Problem

To analyse functional group present in given mono-functional organic compound(s)

Educational Purpose

By performing this lab, the student will learn to distinguish between:

1. Aromatic and non-aromatic compound,
2. Acidic, Basic and neutral functional groups
3. Characteristic tests
4. How compounds are analysed?

Tutorials

 Functional groups

What To Turn In

1. Questionnaire: prelab and postlab

1. Introduction

Many organic compounds contain other atoms besides carbon that contribute significantly to the
physical and chemical properties of the compound. Heteroatoms are atoms other than carbon or
hydrogen and the groups they form, functional groups. A functional group is the atom, or
atoms, that are the center of reactivity of a molecule. Although there are a wide variety of
organic compounds, most of them are composed of the elements from the upper right hand
portion of the periodic chart: C, H, N, O, S, Halogen. These compounds can be categorized by
certain structural and reactive features, dictated by the way carbon bonds to itself or another
element, e.g. carbon double bonded to oxygen. Such a grouping of compounds provides us with
the concept of chemical families. These special bonding arrangements have different reactivities
or functions and associated with each family is particular functional group.
Rigorously speaking the functional group is not the whole molecule but only that collection of
atoms that provides a specific chemical function. For example, the chemical family of alcohols is
characterized by the function of the hydroxyl (OH) group, and aldehydes, ketones are
characterized by carbonyl groups (C=O). There are various combinations of hydroxyl groups in
molecules along with carbonyl groups and these combinations can lead to hydroxyketones and
aldehydes (the basis for sugars) as well as to carboxylic acids. Indeed, esters, and amides also
have carbonyl groups (C=O) but differ in their combination with an additional structural feature.
To understand the reactivity of chemical families one must consider the interplay between the
various structural features in that molecule, and identifying the fundamental functional groups is

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a good place to start. To identify the functional groups present in given organic compound is
subjected to following examination systematically.

2. Practical Notes:

Before outlining the general scheme, please consider following points which are of practical
importance.

(a) Quantities of substance for tests. For most tests about 0.1 g solid or 0.1 - 0.2 mL (2 - 3 drops)
of liquid material (NOT MORE) should be used. In general, lower qunatities of substance gives
the most satisfactory results. Students should not waste much time and material by taking too
large quantity of substance for analysis.

(b) Reagents likely to be met within organic analysis are on the reagent shelves. Students are
advised to develop a general knowledge of the physical characteristics of common organic
compounds. If in doubt about the expected result of a test between a certain compound and a
reagent, carry out a trial test with a known compound or blank test (control i.e. everything
without your organic compound) and compare with the unknown.

(d) If a practical book instructs one to use larger quantities (3 - 4 g or more), the quantities
should be scaled down to 1 g or 1 mL of the unknown substance and corresponding quantities of
reagents should be used.

(e) Students are strongly advised against carrying out unnecessary tests, since not only are they a
waste of time but also increase the possibility of error. Thus it is pointless to first test for alcohol
or ketone in a basic compound containing nitrogen! Instead tests for amines, etc. should be done
on such a compound.

(f). A systematic approach cannot be over emphasised in group classification tests to avoid
confusion and error.

3. Preliminary examination
It provides useful information about the given organic compound and it includes

3.1 Physical state: solid (generally, higher molecular weight carboxylic acid, aromatic phenols,
amides) or liquid (generally, lower molecular weight carboxylic acid, alcohol, ether, ester,
aldehyde, ketone)

3.2 Colour:

Observation Inference
Colourless Carbohydartes, aldehydes, ketones, carboxylic
acids, alcohols, esters
Colourless turning reddish or pinkish (due to Phenols, amines, aminophenols

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slow oxidation in air)
Reddish orange Nitroamines, azpo compounds, napthaquinones
Yellow Nitro compounds, quinines, iodoform
Greenish yellow p-Nitoso compounds

Note: The above information is useful if given organic compound is pure. Sometimes colour is
due to impurity/ies present in the compound.

3.3 Odour: Some organic compounds have characteristic odour

Observation Inference
Bitter almond odour Nitrobenzene, benzaldehyde, benzonitrile
Cinnamon like Cinnamaldehyde
Pleasant fruity Esters
Sweet odour Chloroform, alcohols
Phenolic Phenols
Irritating pungent Lower acids, acid halides, aldehydes
Vinegar like Acetic acid
Fish like Aliphatic amines

Note: Do not inhale vapours of compound directly

3.4 Flame test: Heat the organic compound in a spatula over a low flame. Note the colour of
flame and any odour coming out of it.

Observation Inference
Non-sooty flame Aliphatic compounds
Sooty flame Aromatic compounds
Ammoniacal odour Urea, thiourea etc.
Charring with sweet smell Carbohydrates
/burnt sugar

Note: Some aliphatic compounds like chloroform, carbon tetrachloride etc. produces sooty
flame.

3.5 Solubility test: To a given compound (0.1g/0.2mL) add the corresponding reagent in which
solubility to be determined. Experiment to be performed in a systematic manner as given in
table, it helps to predict the nature of functional group.

Reagent and Test Class Group of Compounds

Soluble in cold or hot water. Lower members of series.
Neutral, acidic or basic. (Test
(If the unknown is soluble do Neutral, e.g. alcohols; Acidic,
with litmus or universal
NOT perform solubility tests e.g. acids, phenols; Basic, e.g.
indicator paper)
below) amines
Aqueous solution is acidic to
Phenols, acids
litmus
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Aqueous solution is alkaline to
Amines
litmus
Aqueous solution is neutral to Aliphatic alcohols, aldehydes,
litmus ketones, esters
Soluble in dil. HCl Basic Most amines
Soluble in dil. NaOH Acidic Most acids, most phenols.
Most carboxylic acids,
Soluble in NaHCO3 Strongly acidic
polynitro phenols
Hydrocarbons, nitrohydro-
carbons, alkyl or aryl halides,
Insoluble in water, acid and
Neutral esters and ethers. Higher
alkali
molecular weight alcohols,
aldehydes and ketones

Organic Compound

water

insoluble soluble

6M NaOH test with pH paper

insoluble soluble acidic basic

6M HCl neutral

insoluble soluble
carboxylic carboxylic amines
acids acids (low MW)
(high MW) (low MW)
aldehydes
ketones
alcohols
aldehydes amines
(low MW)
ketones (high MW)
alcohols
(high MW)
or
alkanes
alkenes
alkyl halides
(low or high MW)

Figure 1. Solubility test flowchart

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4. Detection of extra elements: The elements C, H and O are assumed to pbe present in an
organic compound. N, X= Cl, Br, I and S may also be present in an organic compound. The
presence of theses extra elements may be confirmed by Lassaigne’s test.
Note: This you will not be performing in lab

5. Detection of Unsaturation

5.1 Bromine Test for Multiple Bonds (Alkenes/alkynes): Bromine readily adds across the
carbon-carbon double bond of an alkene to produce a dibromoalkane. A red solution of bromine
in methylene chloride will quickly become colorless as the addition reaction with the alkene
takes place. The general equation describing the test is:

Procedure: Dissolve organic compound (0.1 g/ 0.1 mL) in methylene chloride (2mL) and add
dropwise with shaking a solution of bromine in methylene chloride (5%, 2-3 drops). Discharge
of bromine colour without evolution of HBr shows the presence of unsaturation in given organic
compound

Note: Dense white fumes when a glass rod dipped in ammonia solution is brought near mouth of
test tube if HBr is evolved in the reaction. If this is happening, suggesting electrophilic aromatic
substitution which happens in aromatic compounds with electron donating groups present in
benzene ring. The white or yellow precipitate is due formation of 2,4,6-tribomophenol/ aniline
derivative. E.g. phenols, aniline etc.

5.2 Baeyer Test for Multiple Bonds (alkene/alkynes): This test is positive for double and triple
bonds but not for aromatic rings. It depends on the conversion of the purple ion MnO 4- to a
brown precipitate of MnO2 following the oxidation of an unsaturated compound.

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Procedure: Dissolve organic compound (0.1 g/ 0.1 mL) in water or acetone(if compound is
insoluble in water) (2mL) and add dropwise with shaking a aqueous solution of potassium
permanganate (2%, 2-3 drops). Discharge of purple colour shows the presence of unsaturation in
given organic compound

Note: Other easily oxidized compounds also give a positive test with potassium permanganate
solution. These include aldehydes, some alcohols, phenols, and aromatic amines.

6. Detection of functional groups

6.1 Carboxylic acids:

(a). Litmus test: Because of their acidic nature, carboxylic acids turn blue litmus to red

Procedure: Add blue litmus solution (1 drop) to an aqueous solution of given organic compound
(1 mL). Appearance of red colour suggests presence of carboxylic group.

(b). Sodium bicarbonate test: Sodium hydrogen carbonate reacts with carboxylic acids to give
the sodium salt of the acid and liberates carbon dioxide (brisk effervescence). If the acid is
insoluble in water and the reaction is sluggish dissolve the acid in methanol and add carefully to
a saturated sodium hydrogen carbonate solution, when a vigorous effervescence will be
observed.

R-CO2H + NaHCO3  RCO2-Na+ + CO2 + H2O

Procedure: To a saturated solution of sodium bicarbonate in water (1 mL) add the given organic
compound (a pinch). Effervescence suggests presence of carboxylic group.

(c). Fluorescein Reaction (Dicarboxylic acids):

The anhydrides of aromatic 1,2-dicarboxylic acids on heating with resorcinol gives a dye
fluorescein. This dye in NaOH solution gives a yellowish red solution with green fluorescence.

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Procedure: In a dry test tube, mix a small amount (0.05g) of dicarboxylic acid (such as Succinic
acid or phthalic acid) with roughly double the amount of resorcinol (0.1g) and about 2 to 3 drops
of concentrated H2SO4. Gently fuse on a small flame. Pick up the fused mass at the tip of a
glass rod and dip into dilute NaOH solution (2 mL). A red dye with strong green fluorescence
appears.

6.2 Phenols

(a). Solubility: If compound acidic and soluble in NaOH and does not produce CO2 on treatment
with NaHCO3 solution

(b). Bromine water: Phenols are generally highly reactive towards electrophilic reagents and are
readily brominated by bromine water. e.g.

Procedure: Dissolve or suspend about 0.05 g of the compound in dilute hydrochloric acid (2
mL) and add bromine water dropwise or bromine in glacial acetic acid until the bromine colour
remains. A white precipitate of the polybromo phenol may form. Solid bromophenol derivatives
can be used for the confirmation of the structure of a phenol.

(c). Ferric chloride test: Most phenols react with iron (III) chloride to form coloured complexes.
The colours vary - red, purple, blue or green - depending on various factors, e.g. the phenolic
compound used, the solvent, concentration. Since some phenols do not give colours, a negative
test must not be taken as significant without supporting information.

6 PhOH + FeCl3  H3[Fe(OPh)6] + 3HCl

Procedure: Dissolve 0.05 g of the compound in 2 mL water (or a mixture of water and ethanol if
the compound is not water-soluble) and add an aqueous solution of neutral ferric chloride drop
wise (0.5 mL). Observe any colour changes which may occur.

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Note: A very dilute solution of organic compound is required.

6.3 Alcohols

(a). Ceric ammonium nitrate test: Alcohols replace the nitrate ion in ceric ammonium nitrate,
change the colour of the solution from yellow to red.

(NH4)2[Ce(NO3)6] + ROH  (NH4)2[Ce(OR)(NO3)5] + HNO3

Procedure: Dissolve given organic compound (10 mg) in water (1mL) and add ceric
ammonium nitrate solution (2-3 drops). Immediately observe the colour change. Appearance of
red colour shows the presence of alcoholic group.

(b). Lucas test: The tests for the hydroxyl group not only detect the presence of the group, but
may also indicate whether it is primary, secondary or tertiary.

RCH2OH + HCl + ZnCl2  No reaction

R2CHOH + HCl + ZnCl2  R2CHCl+ H2O

R3COH + HCl + ZnCl2  R3CCl+ H2O

Alcohols are soluble in Lucas reagent but alkyl chorides are insoluble.

Procedure: Add Lucas reagent (2-3mL) to the given organic compound (1mL) in a test tube.
Shake well and note the time required for separation of two distinct layers.

Tertiary alcohol Layers separate immediately

Secondary alcohol Layers separate takes 5-10 mins
Primary alcohol A clear homogeneous solution is obtained

6.4 Aldehydes and ketones

(a). 2,4-Dinitrophenylhydrazine (Brady's reagent) A test for the carbonyl group (>C=O) in
aldehydes and ketones. 2,4-Dinitrophenylhydrazine gives sparingly soluble yellow or red 2,4-
dinitrophenylhydrazones with aldehydes and ketones.

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Procedure: Add 2,4-DNP reagent (3 mL) to 2 drops of the compound in a test tube and shake. If
no precipitate forms immediately, warm on water bath and allow to stand for 5 - 10 minutes. A
crystalline precipitate indicates the presence of a carbonyl compound.

(b). Iodoform test for Methylketones (CH3CO-): This test is used to identify methyl ketones
(ketones with a methyl group on one end). It uses the haloform reaction, in which the methyl
ketone is oxidized to a carboxylic acid and iodoform, a yellow solid, is produced.

Formation of solid iodoform (yellow) is a positive test. (Iodoform can be recognized by its odor
and yellow color and, more securely, from the melting point 119-123oC).

Procedure: Dissolve 0.1 g (or 5 drops) of the compound in 2 mL of water; if it is insoluble in

water add sufficient dioxane to produce a homogeneous solution. Add NaOH (5%, 2 mL)
solution and then introduce the potassium iodide - iodine reagent dropwise with shaking until a
definite dark colour of iodine persists. Allow to stand for 2 - 3 minutes; if no iodoform separates
at room temperature, warm the test tube in a beaker of water at 60oC. Add a few more drops of
the iodine reagent if the faint iodine colour disappears. Continue the addition of the reagent until
a dark colour is not discharged after 2 minutes heating at 60 oC. Remove the excess of iodine by
the addition of a few drops of dilute sodium hydroxide solution with shaking, dilute with an
equal volume of water, and allow to stand for 10 minutes. The test is positive if a yellow
precipitate of iodoform is deposited.

Note: The reaction is given by acetaldehyde and simple methyl ketones. Alcohols containing the
CH3CHROH group (ethanol) will be oxidised under the reaction conditions and also give a
positive test.

6.4.1 Aldehydes including some carbohydrates only (reducing properties)

(a) Fehling's test: Fehling's is always prepared fresh in the laboratory. It is made initially as two
separate solutions, known as Fehling's A and Fehling's B. Fehling's A is a blue aqueous solution
of copper(II) sulfate, while Fehling's B is a clear solution of aqueous potassium sodium
tartrate (also known as Rochelle salt) and a strong alkali (commonly sodium hydroxide).
Equal volumes of the two mixtures are mixed to get the final Fehling's solution, which is a deep
blue colour. In this final mixture, aqueous tartrate ions from the dissolved Rochelle
salt chelate to Cu2+ (aq) ions from the dissolved copper(II) sulfate, as bidentate ligands giving
the bistartratocuprate(II)4- complex as shown below. The tartarate ions, by complexing copper
prevent the formation of Cu(OH)2 from the reaction of CuSO4.2H2O and NaOH present in the
solution.

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Aldehydes reduce Fehling's solution to yellow or red copper (I) oxide and get itself oxidized to
carboxylate ion.

Procedure: Add 2 drops (or 0.05 g) of the compound and Fehling's solution A (1mL, aqueous
alkaline potassium tartrate) and Fehling's solution B (1mL, Copper sulphate solution) and heat
the test tube on a boiling water bath for 3 - 4 minutes. Appearance of red precipitate suggest
aldehyde group presence.

Note: The test is positive for aliphatic aldehydes, but is often indecisive for aromatic aldehydes.

(b) Tollen's reagent (Ammonical silver nitrate solution): Aldehydes are readily oxidised to
carboxylic acids and will reduce Tollen's reagent to produce a silver mirror on the inside of a
clean test tube.

Procedure:
Note: First clean up a test tube with a little hot nitric acid (fume cupboard) and rinse with
distilled water.
Preparation of the reagent: To silver nitrate solution (1 mL) add a few drops of sodium
hydroxide. Then add dilute ammonium hydroxide dropwise until the precipitate just dissolves.
Add 2 - 3 drops of the compound in methanol to Tollen's solution (2 - 3 mL) contained in a very
clean test tube. If no reaction takes place in the cold, warm gently in a water bath. Formation of
silver mirror or a black precipitate is a positive test for aldehyde group.
Note: After the test, pour the contents of the test tube into the sink and wash the test tube with
dilute nitric acid. Any silver fulminate present, which is highly explosive when dry, will be
destroyed.

6.5 Amides (-CONH2)

(a) NaOH test: Amides are decomposed by NaOH to evolve ammonia.

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 Amide Sodium salt of carboxylic acid Ammonia

Procedure: To 0.2 g of compound add aq. NaOH (1mL) and heat the test tube on a burner gently
for 1 - 2 minutes. Show either a moist glass rod with dil. HCl or moist red litmus paper to the
neck of test tube. The evolved gas gives either dense white fumes or turn red litmus to blue.

(b) 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. Biuret is a
condensation compound of urea, equivalent to two molecules of urea less one of ammonia. This
biuret in alkaline medium gives a violet colour with a drop of copper sulphate solution.

Biuret

The term biuret can also apply to the functional group and class of organic compounds with the
general structure RHN-CO-NR-CO-NHR where R is an organic residue.
The Biuret reagent is made of potassium hydroxide (KOH) and hydrated copper(II) sulfate,
together with potassium sodium tartrate. The reagent turns from blue to violet in the presence
of proteins, blue to pink when combined with short-chain polypeptides, violet for urea.

Procedure: To 0.2 g of compound was heated on a flame for 1 min. Upon cooling add aq.
NaOH (1mL) and few drops of copper sulphate. A bluish violet color suggest positive test for
urea.

(c) 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.

Aromatic amide Sodium salt of Aromatic Acid Aromatic acid

Procedure: To 0.5 g of compound containing antibumping granule, add aq. NaOH (20%, 10
mL) and heat the test tube on a burner with gauze wire gently for 10 - 15 minutes. Heat the
contents till there is no evolution of ammonia (till no colour change of red litmus paper). Upon
cooling, add aq. HCl and a white ppt. due to corresponding acid is obtained.

6.6 Primary Amine (-NH2)

Amines undergo a variety of reactions with nitrous acid (HNO2) depending on whether the
amine is primary, secondary or tertiary and whether the amine is aliphatic or aromatic. The
reaction of primary amines (either aliphatic or aromatic) with nitrous acid results in the
formation of a diazonium salt. The diazonium salts of primary aliphatic amines are unstable and
quickly decompose to give carbonium ions and N2, with the gas rapidly bubbling out of solution.
The diazonium salts of primary aromatic amines, on the other hand, are stable in aqueous

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solutions at temperatures or 0-5 C°, at higher temperatures they decompose with loss of N2. If
the aromatic diazonium salts are kept at low temperatures and treated with a nucleophile such as
a phenol, they will form colored azo dyes. When naphthol is added to the suspected diazonium
salt, the formation of a red color or precipitate is indication of the formation of an azo dye.

Diazotisation:

At low temperature (0-5oC) aromatic primary amines dissolved in strong acids (HCl & H2SO4)
reacts with nitrous acid (NaNO2 +HCl) to form water soluble diazonium salts.

Aliphatic primary amines do not form stable diazonium salts under similar condition. They react
with nitrous acid to yield alcohols and nitrogen (causes rapid foaming).

Azo dye formation for aromatic primary amines:

Aryldiazonium salts react with aromatic rings of phenols to form highly coloured azo
compounds. These reactions are called coupling reactions.

Procedure: In one test tube, labeled A, dissolve 0.5 mL g of aniline in 2-3 mL dil HCl and cool
in ice bath at 0-5oC. In test tube B, dissolve 0.3 g of sodium nitrite in 2-3 mL of distilled water
and cool in ice bath at 0-5oC, In test tube C, dissolve 0.2 g of 2-Napthol in aqueous KOH in 2-3
mL and cool in ice bath at 0-5oC. Note: Low temperature is required for this reaction, so check
the temp. before mixing. Now, add contents of B to A and cool again before addition of contents
C to AB mixture. An orange red shows the presence of aromatic primary amine.

Prelab:
1. Why carboxylic acid give brisk effervescence with saturated sodium carbonate solution while
phenol and alcohol does not? Draw an equation to represent this reaction.
2. How will you distinguish propan-2-one and pentan-3-one? Draw an equation to represent this
reaction.
3. Draw the reaction between cyclohexene and bromine in CCl4. What type of reaction is this?
4. What type of reaction is the reaction between an alkyl halide and alcoholic silver nitrate?
How would you expect the structure of the alkyl halide to affect the rate of this reaction?

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7. Identification of known Organic Compounds (perform tests on organic compound batch
given, 4 samples given/4 students, individually work on one compound/person)

Carefully record your observations. Accurately describe each positive test and include factors
such as reaction time, color change, precipitate formation, and the need for heating, stirring, or
shaking.

8. Preliminary tests
8.1 Solubility Tests
Indicate which compounds are soluble in water, and for those that are soluble, indicate
whether the solution is acidic, neutral, or basic.

Record in your notebook

(a) Name of students in Batch__1._________2.__________3._________4.__________

(b) Number on Compounds given: ___________

(c) Preliminary tests (in order as indicated in text)

(d) Respective tests performed (in order)

Experiment Observation Inference

Compound “ X”

Physical Characteristics

State

Colour

Odour

Flame test

Solubility

Cold water

Hot water

Dil. HCl

Aq. NaOH

Test for Unsaturation

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Bromine water test

KMnO4 test

Functional Group Detection

etc.

Result:

The Given Organic Compound

No. __ is acidic/basic/neutral, aromatic/aliphatic contains __________functional group

No. __ is acidic/basic/neutral, aromatic/aliphatic contains __________functional group

No. __ is acidic/basic/neutral, aromatic/aliphatic contains __________functional group

No. __ is acidic/basic/neutral, aromatic/aliphatic contains __________functional group

Post-lab:
1. Can you suggest reason why lower molecular weight alcohol is soluble in water?
2. Why resorcinol is not highly soluble in water but dissolves quickly in aqueous NaOH and
gives dark color?
3. Suggest two strategies for separation of a mixture of aniline and benzoic acid in lab.

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