NEW EXPERIMENTS CHEMISTRY 1

NEW EXPERIMENTS CHEMISTRY

Experiment - 1

Preparation of Some (a) Lyophilic Sols and (b) Lyophobic Sols

Aim : To prepare some (a) lyophilic sols and (b) lyophohic sols.

Theory
Lyophilic sols are directly prepared by adding and shaking the substances that form dispersed phases
with suitable liquids that form dispersion media. Here, attraction between dispersed particles and dispersion
medium are responsible for the stability of colloidal state. Lyophobic sols are not obtained by simple mixing
and shaking of the substances that form the dispersed phase and the dispersion medium. Special methods
are adopted for the preparation of lyophobic sols.

Sols to be prepared
Lyophilic sols: egg albumin sol, starch sol and gum sol. Lyophobic sols: Ferric hydroxide sol,
aluminium hydroxide sol and arsenious suiphide sol.

Apparatus and chemicals required

Beakers (250 mL), watch glass, porcelain dish, measuring jar (100 mL), pipette (10 mL), graduated
pipette (20 mL). starch (500 mg), gum (500 mg), egg (1), sodium chloride (5 g), aluminium chloride (2 g), ferric
chloride (2 g) and arsenious oxide (0.2 g).

Procedure

(A) Preparation of lyophilic sols

1. Preparation of starch sol: Take 100 mL of distilled water using measuring jar and transfer it to a 250
mL beaker. Heat the beaker until the distilled water in ft boils. Make a paste of starch with 500 mg of it
in water. Transfer this starch paste slowly to boiling water in the beaker with oi1stau stoT1nL.
Continue the stirring and boiling of water for atleast 10 minutes after adding the starch paste. Thus the
starch sol is formed.

Note: The gum sol also is prepared by the same procedure with 500 mg of gum instead of 500 mg of
starch.

2. Preparation of egg albumin sol

(i) Take 5 g of sodium chloride in a 250 mL beaker and add to it 100 mL of distilled water using a
measuring jar. Prepare a clear solution by dissolving NaCl in distilled water.
(ii) Break an egg into a porcelain dish. Pipette out the albumin into the above sodium chloride
solution while stirring thoroughly. Egg albumin sol is formed.

(B) Preparation of lyophobic sols

1. Preparation of ferric hydroxide sol

(i) Take 100 mL of distilled water in a 250 mL beaker using 100 mL measuring jar and boil the
water. Add 2 g of ferric chloride powder to this boiling water while stirring well.
(ii) Take another 100 mL of distilled water into another 250 mL beaker and boil it.
(iii) Take 10 mL of ferric chloride solution prepared in step (i) and add it to boiling water prepared
instep (ii) drop by drop with vigorous constant stirring. Keep the water boiling. A brown ferric
hydroxide (Fe (OH) 3) sol is formed.

Note: Adopt same procedure to prepare Al(OH)2 sol as given for the preparation of Fe{OH). sol. Ai(OH)3 sol
is white in colour.

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 NEW EXPERIMENTS CHEMISTRY 2
CHROMATOGRAPHY
Chromatography is essentially a separation process for mixture of substances, but this is also
adapted to identify the components of mixtures. Purification of the components is also possible with this
method. Chromatography is a physical process in which the components to be separated are distributed
between two phases, one of which is stationary while the other is a mobile phase.

The stationary phase is usually in the form of a packed column in column chromatography, a thin
layer adhering to a suitable form of backing material such as glass in thin-layer chromatography. In column
chromatography, mobile phase flows through the packed column, while in thin layer chromatography, mobile
phase moves by capillary action. In thin layer chromatography, stationary phase may be either a liquid or a
solid and the mobile phase may be a liquid or a gas.

In partition chromatography, stationary phase thin film of liquid adsorbed on an essentially inert
support Mobile phase ma le a liquid or a gas. Paper chromatography is an example of partition
chromatography in which liquid present in the pores of paper is stationary phase and some other liquid is
mobile phase. Separation depends upon partition of substance between two phases and the adsorption
effects of inert support (paper) on compounds undergoing chromatographic separation. In chromatography,
substance equilibrates between a mobile and a stationary phase. The more the interaction of substance with
the stationary phase, slower is its movement. Separation of the components of a mixture by using paper
chromatography is given below.

Experiment: 5.1

Separation of Pigments Present in the Extract of Leaves and Flowers

Aim: Separation of pigments present in the extracts of leaves and flowers by paper chromatography and
determination of Rf value of components. Select the leaves and flowers whose pigments are soluble
in water.

Theory

Paper contains lots of pores, Water molecules which enter into the pores of filter paper act as the
stationary phase. Flowers and leaves are selected in such a way that their pigments are soluble in water. The
moving phase is solvent water. As the moving phase passes through the spot (the reference line) on which
the sample (extract from flower or leaf) has been adsorbed, the solvent dissolves the components more or
less readily: depending upon the solubility and carries them along with it while moving on the support. If the
pigments are not soluble in water, solvents like hexane, toluene, acetone or a mixture of solvents such as
methanol-water can be used as mobile phase. For a given solvent or solvent mixture, at constant
temperature, it is possible to determine the characteristic rate of movement of each substance on the
chromotographic paper, as the moving phase moves. This is represented by relative front or retardation factor
also called Rf value. Rf values of different compounds are different even if the mobile phase (solvent) is same.
Further more, Rf value of a compound may be different in different solvents. Rf values are calculated by using
the following equation.
Distance travelled by the substance from reference line (cm)
Rf 
Distane travelled by the solvent front from reference line (cm)

Since solvent front moves fa4ter than the compounds, the Rf value of a substance will always be less
than one. Rf value has no unit. If the compounds are coloured, then their position on the chromatographic
paper may be easily located. However, if the compounds are colourless, they may be treated with a reagent,
which imparts characteristic colour to them. This reagent is given the name developer. Iodine is the most
commonly used developer in paper chromatography.

Materials required

Whatman’s filter paper No.1, glass jar or, 100 mL beaker, test tubes, flower extract or extract of
leaves, distilled water, methanol, acetone, chloroform.

Procedure

1. Grind flowers/leaves in a mortar and transfer the paste into a test tube. Add small amounts of water,
methanol or acetone t the crushed material. Shake well. Filter and collect the filtrate in a test tube.

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 NEW EXPERIMENTS CHEMISTRY 3
2. Cut Whatman’s filter paper No.1 into a rectangular shape of 4 cm x 16 cm and mark a line 3 cm from
one of the ends of the filter paper with the help of a pencil [Fig.5. 1(a)].
3. Using a capillary glass tube, put one spot ‘A’ with the flower extract. Allow the spot to dry.
4. Using a glass rod and cellotape, hang the filter paper in a jar containing water (or 19 mL petroleum
ether, boiling range 60-80°C and 1mL of chloroform or a mixture of petroleum ether, boiling range 60-
80°C and acetone in the ratio of 9:1), so that the solvent does not touch the reference line as shown
in fig.5. 1(b)
5. Carefully observe the filter paper in the jar until the mobile phase (solvent) rises upto 2/3 of the length
of the paper [Fig. 5.1(c)].
6. Take the filter paper out, mark the solvent front, outline the spots with the help of a pencil and allow
the paper to dry.
7. Measure the distance of the solvent front and the centre of different spots with respect to the
reference line as shown in fIg.5. 1(d).
8. Find cut the number of pigments present in the extract of flowers/leaves.
9. Calculate the Rf values of different spots as explained earlier.
10 . Record your observations in a table.

Table 5.1: Separation of pigments of flowers / leaves

Distance of Distance of the

the spot from solvent front
[Link]. Colour of the spot Rf value
reference line from reference
(in cm) line (in cm)
1. Green (Chlorophyl) 5.1 cm 9.8 cm 0.52

2. Yellow (Xanthophyl) 7.2 cm 9.8 cm 0.73

3. Red (Carotone) 9.7 cm 9.8 cm 0.99

A 5.1
For Green = R f    0.52
X 9.8
A 7.2
For Yellow = R f    0.73
X 9.8
A 9.7
For Red = R f    0.99
X 9.8

Result : Rf values of components of flower / leaf are ………………………….

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 NEW EXPERIMENTS CHEMISTRY 4
Precautions

1. Dip the filter paper in the solvent in such a way that the spot of the mixture should not touch the
solvent directly.

2. Do not allow the extract spots to spread. Use capillary glass tube to put the spot on the paper.

Experiment: 5.2

Separation of Ions Pb2+ and Cd2+ in a Mixture of Compounds Using Chromatographic Technique

Aim: Separation of ions Pb2+ and Cd2+ in a mixture of compounds using chromatographic technique.

Theory

Principle for the separation of cations is same as given in experiment 5.1. The two cations to be
separated are colourless, therefore, a developer is needed. In the present: case, ammonium suiphide
(NH4)2S, can be used to locate the position of these ions on chromatographic paper. (Ammonium suiphide is
prepared by passing H2S gas through 10% liquor ammonia solution in water for 45 minutes).

Materials required

Whatman’s filter paper No. 1, glass jar, test tubes, 1-2% solution of Pb(NO3)2 and Cd(NO3)2, ethanol,
6.0 M HNO3

Procedure

1. Cut the Whatman’s filter paper No.1 in a rectangular size of 4 cm x 16cm

2. Using a capillary glass tube put a spot of the mixture on the marked reference line.

3. Using a glass rod and cellotape, hang the filter paper in a jar containing a mixture of ethanol, 6.0 M
HNO3 and distilled water in the ratio of [Link].

4. Keep the jar undisturbed till the mobile phase (solvent) rises upto two thirds of the length of the paper.

5. Take the filter paper out from the jar and mark the solvent front.

6. Spray ammonium sulphide solution on the chrornatographic paper to see yellow and black coloured
spots. Mark the position of the spots with a pencil and allow the filter paper to dry.

7. Measure the distance of the solvent front and the different spots of the cations with respect -to the
reference line. Record the observations in a table. Calculate the Rf values for each cation (Pb2+ and
Cd2+).

Table 5.2: Separation of Pb2+ and Cd2+ ions by paper chromatography

Distance of the Distance of the
component from solvent front from
[Link]. Colour of the spot Rf value
reference line reference line
(in cm) (in cm)
1. Cd+2 2.0 cm 3.5 cm 0.57
2. Pb+2 3.3 cm 3.5 cm 0.94

Calculation :
A 2.0
For Cd+2 = Rf    0.57
X 3.5
A 3.3
For Pb+2 = R f    0.94
X 3.5

Results : Rf values of components of Pb2+ and Cd2+

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 NEW EXPERIMENTS CHEMISTRY 5
Precautions

1. Dip the filter paper in the solvent in such a way that the spot of the mixture should not touch the
solvent directly.

2. Do not allow the extract spots to spread Use capillary glass tube to put the spot on - the paper.

Questions for discussion

1. How is adsorption phenomenon applied in the separation of components in a mixture by

chromatography?
2. What is the criteria of a developer?
3. What is a chromatogram?
4. What is meant by the term Rf value?
5. Name different chromatographic techniques.

CHARACTERISTIC TESTS FOR CARBOHYDRATES AND PROTEINS

9.1 Carbohydrates

Carbohydrates are optically active polyhydroxy aldehydes or polyhydroxy ketones or their derivatives.
They have the general formula Cx(H2O). They are classified into monosaccharides, disaccharides,
oligosaccharides and polysaccharides.

Monosaccharides
They are reducing sugars. They can reduce Tollen’s reagent and Fehling’s solution.
Ex: Glucose, fructose etc.

Oligosaccharldes
They yield 2-10 monosaccharides on hydrolysis. They are non-reducing sugars.
Ex: Sucrose, maltose etc.

Polysaccharldes
They contain more than 10 monosaccharide units. Their general formula is (C6H10O5)
Ex: Starch, cellulose etc.

The carbohydrates perform two important functions.

a) They act as bio-fuels to provide energy for the functioning of living organisms.
b) They act as constituents of cell membrane.

9.2 Qualitative Tests for Carbohydrates

Experiment Observation Inference

Cone. H2SO4 test: Take. small Charring takes place,
Indicates the presence
1. quantity of given compound then add burnt sugar smell is
of carbohydrate.
few drops of conc. H2SO4 and heat. observed.
Molisch Test: Take 1-2 mL of
aqueous solution of carbohydrate
A deep violet ring is
and add few drops of 10% alcoholic Indicates the presence
2. formed at the junction of
solution of alpha-naphthol. Add of carbohydrate.
the two liquids.
cone H2SO4 slowly along the sides of
the test tube.
BenedIct’s test: Take 1-2 mL of
aqueous solution of carbohydrate in Brick red precipitate is
Presence of reducing
3. a test tube arid add 1-2 mL of observed due to
sugar is indicated.
Benedict’s reagent. Keep the test formation of Cu2O.
tube in a boiling water bath.
Tollen’s test : Take 2-3 rnL of
aqueous solution of carbohydrate in
Presence of,reducing
a test tube and add 2-3 mL of Tollen’s A shining silver mirror
4. sugar is indicated.
rea,gent. Keep the test tube in a is observed,
.
boiling water bath for about 10
minutes.

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 NEW EXPERIMENTS CHEMISTRY 6
9.3 Proteins

Proteins are high molecular mass, long chain polymers -composed of a-aminoacids. The linkage
between two amino acids. is called peptide linkage. The product formed by the combination of two a-amino
acids is called dipeptide and with three amino acids is a tripeptide. A polypeptide contains large’number of a-
amino acid molecules. A polypeptide having molecular mass greater than. 10.000 is coiled protein. Proteins
are constituents of cells and hence are present in all living bodies. Some of the proteins act as enzymes arid
catalyse the biochemical reactions. Most of the hormones which control various metabolic processes are
basically proteins. Some proteins act as antibodies and protect the body from the effect of invading
substances.

9.4 Qualitative Tests for Proteins

Experiment Observation Inference

Biuret test: Take about 2 mL of
given sample solution in a test
tube and add about 2 mL of Bluish violet colouration is Indicates the presence of
1.
10% NaOH solution to it. Then, observed. protein.
add 4-5 drops of 2% CuSO4
solution to it.
Ninhydrln Test: Take 2-3 mL of
aqueous solution of given
sample in a test. tube and add Blue colouration is
2. Protein is present.
3-4 drops of Ninhydrin solution observed.
to it. Then heat
the solution.
Xanthoprotlc test: Take about
2 mL of given ‘sample in a test
Yellow precipitate is
tube and add few drops of conc. Protein is present.
observed.
HNO3. Then heat the solution.
3.
An orange colour
Cool the yellow precipitate in a Protein is present.
appears.
test tube and add a few drops of
10 M sodium hydroxide solution.

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