Cagatan, Jay Marie B.

October 16,
2020
Experiment 3
Detection of Functional Groups in Organic Substances

I. Introduction

In certain respects, qualitative organic analysis, an identification of organic compounds

on the basis of their physical and chemical properties, is similar to the identification of
plants and animals on the basis of their taxonomy, structural characteristics and presumed
natural relations. It is relevant to always identify a particular functional group in the
molecules of chemical compounds to identify any organic compound into a given family.

Functional groups affect the physical, chemical, and spectral properties of an organic
compound. By examining such physical properties, analyzing its chemical activity with
various classification reagents, and examining other spectral data, a chemist may define
the functional groups of a compound.

In this experiment it will subject a series of organic compounds to specific chemical

reactions in order to identify which class of functional group the compound belongs to.
During the course of this experiment, there would be a test for saturated and unsaturated
hydrocarbons (Bromine test and Baeyer’s test), test for reactions of Alcohol (The Lucas
test, Iodoform Test and Ferric Chloride test with phenol), test for the reaction of
Aldehydes and Ketones (Schiff’s test and Reduction test), test for the reaction of
Carboxylic Acids (NaHCO3 test), and lastly test for the reaction of Amines ( Nitrous
Acid test). The data will be presented in a tabular form showing the test done and its
result and remarks.
II. Objectives:

At the end of the experiment, the students should be able to:

1. Determine which functional group is present in organic substances using the different
test or reagent.
2. Identify the different ways in detecting functional groups in organic substances.

III. Apparatus& Chemicals

Apparatus Used:
• Test tubes (3)
• Test tube rack
• Pipette
• Glass bottle
• Retort stand
• Bunsen Burner
• Beaker (3)
• Buchnur funnel
• Buchnur flask
• Siphon pump
• Flask
• Dropper
• Graduated cylinder
• Retort stand
• Wire gauze
• Tripod
• Red litmus paper
• Spatula
Chemicals Used:
• Organic Compound
• Dilute Hydrochloric acid
 • Organic Compound
• Sodium nitrate
• B- naphthol
• Distilled water
• Dilute sodium hydroxide
• Primary Amines
• Secondary Amines
• Tertiary Amines
• Sodium Nitrite
• Distilled Water
• Hydrochloric acid
• Sodium hydroxide solution (25%)
• Distilled water
• Benzene sulphonyl chloride
• Concentrated Hydrochloric acid
• Vinegar
• Baking soda

IV. Procedures

Characteristic reactions differentiating saturated aliphatic hydrocarbons from unsaturated

aliphatic hydrocarbons (Unsaturation)

Bromine Test
• Add dropwise solution of bromine water to about 3.0 ml of the sample of organic
solvents. Mix carefully the contents of the test tube. The orange color of bromine quickly
disappears to the moment, when total multiple bonds are saturated.

Baeyer’s Test
 • Add dropwise 0.1 % acetone solution of KMnO4 to about 1 ml of the examined sample.
Mix the solution carefully after the addition of each drop and wait for the disappearance
of the KMnO4 pink color. The quantity of decolorized solution and the speed of its
decolorizing depend on the number of multiple bonds in the molecule of the examined
compound.

Characteristic reactions of alcohols

Luca’s Test
• Add 5 ml of Lucas’ reagent to three dry test tubes with ground-in stoppers, containing
respectively primary, secondary, and tertiary alcohols (0.5 ml). Mix the content for a
while, put them away to the laboratory rack, and check the time difference, necessary in
each case for turbidity and two-phase solution formation.
Iodoform test by Lieben
• 0.5 ml of ethanol solution add about 1 ml of iodine in potassium iodide solution ( er
solution, till yellow color of iodine will dissapear. After mixing and heating the tube
content to 60o C, specific aroma and yellow crystals of iodoform will appear.
Reaction of phenol with ferric chloride
• Add 1 drop of 2% FeCl3 solution to 1% phenol water solution (3 ml). A violet color of
this mixture will form. The color disappears after the addition of some drops of 1 M
sulfuric acid.

Characteristic reactions of aldehydes and ketones

Schiff’s reagent test – detection of aldehydes
• 2 ml of formic aldehyde solution add a few drops of Schiff’s reagent (basic fuchsin in
sodium meta-bisulfite solution) and mix. After a few minutes, a purple (amaranth) color
will appear.
Reduction tests of aldehydes
Fehling’s Test
• In a test tube, mix 1 ml of Fehling I reagent and 1 ml of Fehling II. Heat until boiling
and then add a few drops of formic solution. Mix well and prolong the heating in a water
 bath. Observe the precipitation of the orange-red colored sediment of cuprous oxide
Cu2O.

Tollen’s Test (Silver Mirror Test)

• carefully cleaned the test tube add 1 ml of 0.1 M AgNO3 solution and then start adding
dropwise 2 M water ammonia solution (ammonium base NH4OH). After mixing, the
initial AgOH precipitate will solubilize, forming [Ag(NH3)2]OH complex compound.
Add to the tube 5 drops of glucose solution, mix, and heat tube in a boiling water bath.
Watch the appearance of a silver mirror on the tube walls.

Characteristic reactions of carboxylic acids

Reaction of carboxylic acids with sodium bicarbonate NaHCO3

• Add about 1 ml of 5% baking soda (NaHCO3 solution) to 2 ml of vinegar (acetic acid -
CH3COOH, a short-chain carboxylic acid solution). After shaking, observe the release of
CO2 gas bubbles, gathering on the container walls.
Characteristic reaction of aliphatic amines
Nitrous Test
• Dissolve 2-3 drops of your liquid amine in 2.0 mL of 2.0 M HCl in a small test tube.
Cool to 5-10 °C in an ice bath and add 5 drops of cold 20% aqueous sodium nitrite
solution. The immediate evolution of a colorless gas (nitrogen) indicates a primary,
aliphatic amine.

V. Data & Results

Test for Results Remarks
Saturated & Unsaturated Saturated & Unsaturated
hydrocarbons (Unsaturation): hydrocarbons (Unsaturation
A) Bromine Test Orange-red color disappears A) It shows
decolorization when it
C2H4 + Br2 → C2H4Br2
is shaken with the
colorless
presence of alkene, so
it is unsaturated.
B) Baeyer's Test Pink color disappears B) Decolorization
KMnO4 + H2O → 2KOH + from clear and purple
2MnO2 + 3[O]-C=C- + H2O→-
to cloudy and yellow
C(OH)C(OH)- + KOH
(Colorless) or brown is positive,
also indicates
unsaturation and
presence of alkene

Reactions of Alcohols:

Reactions of Alcohols:
A) Lucas Test Primary: Remains colourless and
A) Presence of
forms an oily layer when heated Alcohol and it is
unsaturated
Secondary: Appearance of
turbidity after an hour
Tertiary: Appearance of turbidity
immediately

B) Iodoform Test Formation of yellow precipitate B) It is saturated with

the Presence of
CHI3 + HCOONa + 5 NaI + 5 ethanol
H2O
yellow precipitate
C) It is saturated with
C) Ferric chloride Test with the Presence of phenol
Formation of stable-colored violet
Phenol
complexes.
Fe(OAr)3 + 3HCl violet complex

Reactions of Aldehydes & Reactions of Aldehydes &

Ketones: Ketones:
A) Schiff’s Test nssnjsjs A) It is saturated and
 ndnsnsns Appearance is of purple coloration has the presence of the
aldehyde group.
B) Reduction Tests:
B.1 Fehling’s Test Formation of red precipitate.
B.1) Fehling’s Test
Cu2O(s) + RCOO- + 3H2O
It is saturated and
red colored precipitate forms.
indicates the presence
of an aldehydegroup.

B.2) Tollen’s Test

Produces silver mirror. It is saturated and
B.2 Tollen’s Test
RCOONH4+2NH3+H2O+2Ag indicates the presence
(Silver Mirror)
of aldehydes.

Reactions of Carboxylic Acids: Produces carbon dioxide gas in the Unsaturated and
(NaHCO3 Test) form of a brisk effervescence indicates Presence of
HCOONa + H2O + CO2 carboxylic acid
(Gas bubbles)

• Unsaturated and
Reaction of Amines: Primary: Evolution of gases
indicates the Presence
(Nitrous Acid Test) ROH + H2O + N2 of Amine

Secondary: Formation of yellow • Saturated and

indicates the presence
oily nitrosoamine
of amine
RN – NO + H2O

Tertiary: Formation of soluble • Unsaturated and

indicates the presence
nitrite salts. of amine.
H2NH+ ONO -

VI. Analysis (Discussion)

Saturated hydrocarbons contain carbon-carbon single bonds are called saturated

hydrocarbons. They are also called Paraffins or Aliphatic Hydrocarbons. They may have straight
chain, branched or ring structure. Unsaturated hydrocarbons that contain carbon-carbon double
bond or triple bond are called unsaturated hydrocarbons.

Tests for Unsaturation

There are two tests for determining unsaturation in an organic compound.
1. Bromine Test
In this test, the orange-red colour of bromine solution disappears when it is added to an
unsaturated organic compound (unsaturated hydrocarbon).

2. Baeyer’s Test (Alkaline KMnO4 Test)

In this test, pink colour of KMnO4 disappears, when alkaline KMnO4 is added to an
unsaturated hydrocarbon. The disappearance of pink colour may take place with or without
the formation of brown precipitate of MnO2.

Tests for Alcoholic Group

Alcohols are compounds in which the hydroxyl group (-OH) is linked to aliphatic carbon chain
or in the side chain of an organic compound. Depending upon the number of hydroxyl group,
alcohols are classified as mono (contain only one –OH group), di (contain two –OH groups) and
trihydric (contains three –OH groups).
1. Lucas Test
Primary, secondary and tertiary alcohols react in different way with Lucas’ reagent, the
solution of anhydrous zinc chloride in concentrated hydrochloric acid. Primary alcohols,
containing less than six carbon atoms, do not react with the reagent, and give clear, slightly
dark solution. With secondary alcohols, turbidity of the solution 1 appears, forming two
phase solution after 1-1.5 hours. In the presence of tertiary alcohols, the solution turbidity
and solution separation for two phases form very quickly. Tertiary alcohols react with
concentrated HCl even in the absence of ZnCl2.

2. Iodoform Test
This test is given by acetaldehyde, all methyl ketones and all alcohols containing CH3-CH-
OH group. When alcohol is warmed with sodium hydroxide solution and iodine, a yellow
precipitate of iodoform is formed.

3. Ferric Chloride Test with Phenols

Ferric chloride reacts with the hydroxyl groups of phenols and phenol derivatives and forms
stable-colored complexes: violet, violet-bluish, blue, green and red. This reaction depends on
the structure and presence of complex-created groups (aldehyde, ketone, carboxylic,
hydroxyl, or sulfonic) located in the orto-position of phenols. Phenols without complex-
created groups in the orto-position react with FeCl3 only in water, forming unstable complex
color compound
Differentiating tests for aldehydes and Ketones:

1. Schiff’sreagenttest–detectionofaldehydes
To 2 ml of formic aldehyde solution add a few drops of Schiff’s reagent (basic fuchsin in
sodium meta-bisulfite solution) and mix. After a few minutes, a purple (amaranth) color will
appear.

2. Reduction tests of aldehydes

These tests reveal reducing properties of aldehydes and carbohydrates. They are based on the
reduction of heavy metal ions and simultaneous oxidation of carbonyl group of aldehydes or
monosaccharides to carboxyl group.

a. Fehling’s test
This is an important test to distinguish aldehydes from ketones. The reagents used in
this test are Fehling’s solution A and Fehling’s solution B. Fehling’s solution A is an
aqueous solution of copper sulphate and Fehling’s solution B is a clear solution of
sodium potassium tartrate (Rochelle salt) and strong alkali (usually NaOH).

The final Fehling’s solution is obtained by mixing equal volmes of both Fehling’s
solution A and Fehling’s solution B that has a deep blue colour. In Fehling’s solution,
copper (II) ions form a complex with tartrate ions in alkali. Aldehydes reduces the
Cu(II) ions in the fehling’s solution to red precipitate of cuprous oxide(copper (I)
oxide).
 b. Tollen’s Test
Tollen’s reagent is ammoniacal silver nitrate. Aldehydes react with Tollen’s reagent
to form elemental silver, accumulated onto the inner surface of the reaction vessel,
producing silver mirror on the inner surface of the vessel.

Tests for Carboxylic group

Carboxylic acids are organic compounds containing carboxyl functional group. It is of two
types aliphatic and aromatic. Aliphatic acids are soluble in water where as aromatic acids are
sparingly soluble in water. Formic acid and acetic acid are the simplest aliphatic acid and
benzoic acid is the simplest aromatic acid. Formic acid and acetic acid are liquids.
Carboxylic acids such as benzoic acid, oxalic acid, phthalic acid, tartaric acid etc are
colourless crystalline solids.

1. Sodium Hydrogen Carbonate Test

Carboxylic acids reacts with sodium hydrogen carbonate to produce carbon dioxide gas
which can be seen in the form of a brisk effervescence.

Tests for Amines

Amines are derivatives of ammonia in which one or more hydrogen atoms are replaced
by alkyl or aryl groups.
When one of the three hydrogen atoms is replaced by alkyl or aryl group, primary amine
is formed. It is generally represented as RNH2.
When two of the three hydrogen atoms are replaced by alkyl or aryl group, secondary
anime is formed. It is generally represented as R2NH.

When all the three hydrogen atoms are replaced by alkyl or aryl substituents, tertiary
amine is formed. It is generally represented as R3N.
1. Nitrous acid Test
Primary aliphatic amines react with nitrous acid to produce nitrogen gas which is seen as
bubbles.

Secondary amines react with nitrous acid to form a yellow oily nitrosoamine.

Tertiary amines react with nitrous acid to form soluble nitrite salts.
VII. QUESTIONS:
1. What is a functional group?
Functional group are atom or group of atoms that replaces hydrogen atoms from
hydrocarbon. They may be –OH, -COOH, -CO, -CHO, -Cl, -COCl, -COOR etc.
Functional groups are responsible for the characteristics of a molecule

2. How can you distinguish between methanol and ethanol chemically?

Methanol and ethanol are distinguished by a type of test known as Iodoform.
When ethanol is warmed with iodine in the presence of NaOH, it forms a yellow
colored precipitate but methanol do not react positively to Iodoform test.

3. How to prepare a nitrous acid solution?

Nitrous acid is prepared by having solid sodium nitrate and cold dilute HCl are
the chemicals you want to prepare nitrous acid. Nitrous acid is a weak monobasic
acid. Solid sodium nitrate (NaNO3) is heated to decompose solid sodium nitrite
(NaNO2) and oxygen (O2) gas. This reaction is an oxidation-reduction reaction.
Solid sodium nitrite is dissolved in water to prepare aqueous sodium nitrite. No
reaction is occurred in this step. Only a dissolving occurs. Add cold dilute HCl to
aqueous sodium nitrite solution. It gives dilute nitrous acid and NaCl solution.

4. What are the significance and the composition of the following reagents: Lucas, Schiff’s,
& Tollen’s?
The Lucas test in alcohols is a test to differentiate between primary, secondary,
and tertiary alcohols. It is based on the difference in reactivity of the three classes
of alcohols with hydrogen halides.
The Schiff’s test is a relatively general chemical test for detection of many
organic aldehydes that has also found use in the staining of biological tissues.
 The Tollen’s test is a qualitative laboratory test used to distinguish between an
aldehyde and a ketone. It exploits the face that aldehydes are readily oxidized,
whereas ketones are not.

5. Name at least two (2) other tests not mentioned in this experiment for functional group
detection.
Other tests that were not mentioned in this experiment are, Biuret reaction, for the
test for amides and action of Borsche’s reagent, for the test for aldehydes or
ketones.

VIII. Conclusion

The chemical reactions used in functional group analysis must be sufficiently selective
with respect to the functional group being analyzed. The content of the functional group
can be measured by determining either the concentration of the products formed or the
consumption of a volatile reagent. A very significant aspect of experimental organic
chemistry is the study and characterization of unknown organic compounds. The findings
of this experiment indicate the analysis and result of the various experiments in order to
identify functional groups in organic substances. With their accompanying observation of
color and how to detect the elements. With respect to the functional group being studied,
the chemical reactions used throughout functional group analysis must be appropriately
accurate.

IX. References
Amrita (2013). Retrieved from amrita.olabs.edu.in., (12 October 2020)Tests for the
functional groups. amrita.olabs.edu.in/?brch=8&cnt=1&sim=141&sub=73
Bhagya. (2018, October 11). Retrieved from https://www.learncbse.in/tests-for-the-
functional-groups-present-in-organic-compounds/
(n.d.). Retrieved from https://www.umb.edu.pl/photo/pliki/WF_jednostki/zaklad-chemii-
medycznej/lab._5.pdf
 (n.d.). Retrieved from http://wwwchem.uwimona.edu.jm/courses/Fehling.html
(n.d.). Retrieved from William Reusch, Professor Emeritus (Michigan State U.),
Functional Groups

VIDEO REFERENCES:
https://youtu.be/vbt_e_FFGhc
https://youtu.be/fcZ29aASk1o
https://youtu.be/yjLB_ntM324
https://youtu.be/biV1tzpUAwU
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https://youtu.be/vBMGNzRYngk
https://youtu.be/s3s3aLx3cfU
https://youtu.be/4yPjkRLcauI
https://youtu.be/iOeDDme-Tl0

X. Documentation