SEPARATING

TECHNIQUES
CHROMATOGRAPHY
LEARNING OBJECTIVES:
Within the session, you should be able to:
 Describe paper chromatography and
interpret chromatogram;
 Explain the use of a locating agent in
the chromatography of colorless
compound.
 CHROMATOGRAPHY
Chromatography is the method of separating two or
more components that dissolve in the same solvent.

Chromatography is a technique of
using a solvent to separate a
mixture into its components.

Chromatography paper
Mixture to be separated

Solvent e.g. ethanol

 CHROMATOGRAPHY

Example, we need to determine the

purity of a food coloring (which may
be made up of different dyes).
 A spot of the food coloring is applied to
the chromatography paper.
 The chromatography paper is dipped in
ethanol (solvent).
 Ethanol travels up the paper, carrying the
dyes along.
 CHROMATOGRAPHY
Note:
A dye that is more soluble in the solvent will
travel further.

Which of the following dyes that has

the highest solubility in the solvent?
A Blue dye
B Yellow dye
 CHROMATOGRAPHY

How do we interpret the result of

chromatography?
The chromatography paper with the
separated components is called a
chromatogram.

The chromatogram shows that the

food coloring is a mixture of two
dyes. It is not pure.
 CHROMATOGRAPHY
How does the chromatogram of a pure
substance look like?

It gives only one spot on a

chromatogram. Thus, it shows
the substance is pure.
 CHROMATOGRAPHY
Solve
nt
What are Rf values? front
Spo
t

The positions of
the solvent front
and spot on a
chromatogram
depend on how
long the
experiment was Chromatogram Chromatogram
after a period after a longer
allowed to run. of time. period of time.
 CHROMATOGRAPHY
The ratio between the distance
travelled by the substance
and the distance travelled by the
solvent is a constant.

This ratio is called the retention

factor or Rf value of the substance.

It will not change as long as

chromatography is carried out
under same condition (same
solvent and temperature)

This property allows us to easily identify a substance on a

chromatogram.
 CHROMATOGRAPHY
E.g. – notice the same Rf values

Rf = 3 cm / 4.5 cm Rf = 3.5 cm / 5.2 cm

= 0.67 = 0.67

Chromatogram after Chromatogram after a

a short time. longer time.
 CHROMATOGRAPHY
How can we identify a banned
substance present in food coloring?

 Identical dyes produce spots at the same height.

 Sample X does not contain the banned dyes A, B and D.
 However, X contains the banned dye C. Therefore, it must
not be consumed.
 CHROMATOGRAPHY
What other conclusions can be
drawn from this chromatogram?

 Dyes A and D are pure.

 Both dye B and dye C are mixtures of two different dyes.
 Sample X is a mixture of three dyes.
 Uses of Chromatography
Chromatography is used to
 separate the components in a sample,
 identify the number of components in a
sample,
 identify the components present in a
sample/ identify the substances
 determine the purity of a sample.
 Locating Agents
 Sometimes the substances being separated are colourless.
 Locating agents can be used to show where the spots are.
 Locating agents bind to the chemicals in the spots.
 Sometimes, another chemical is then added which reacts with
the locating agent to produce a colored spot or the
chromatogram is put under ultraviolet light and the locating
agents glow to show where the spots are.
 KEY IDEAS
 Chromatography is the method of separating two or more
components that dissolve in the same solvent. It can be used to
identify the components of a sample.
 A chromatogram shows the separated components of a sample.
 A chromatography is also used to determine the purity of a
sample. A sample gives only one spot on a chromatogram.
 Rf value = distance travelled by the substance
distance travelled by the
solvent
 A locating agent is used on a chromatogram to make colorless
substances visible. It reacts with the colorless substances to form
colored spots.
 CRYSTALLIZATION
 Crystallisation is used to obtain pure solid from its
solution  specifically used for soluble solids that
decompose during the heating.
 The principle of crystallisation is based on the extent
of solubility of solids in water at different
temperatures.
 The type of mixture to be separated by this method
is soluble solid from a solution.
Eg. Sugar solution, copper (II) sulfate solution
 CRYSTALLIZATION
How is crystallization carried out?

• The solution containing the dissolved solid is

heated until it becomes saturated.
• The hot saturated solution is allowed to cool to
room temperature.
• Pure crystals are formed in the solution after
cooling.(The crystals are removed by filtration. To purify
the crystals, they can be washed with cold distilled water
and dried between filter papers)
CRYSTALLIZATION
 CRYSTALLIZATION
Why is advisable to allow the hot saturated
solution to cool slowly?
 Saturated solution cooled quickly small or
powdery crystals formed
 Saturated solution cool slowly bigger size of
crystals formed
How do we know if a solution is saturated?
 Use a clean glass rod and dip it into the boiling solution.
When the glass rod is cooled, crystals can be seen on the
surface of the glass rod. This shows that the hot boiling
solution has become saturated.
 EVAPORATION
TO DRYNESS
 Evaporation to dryness is used to separate
soluble solid particles by heating the solution until
all water has boiled off  commonly used to
recover salt from seawater.
(Instead of boiling a solution to saturated, the solution is
boiled until all the solvent is evaporated completely, leaving
behind the crystals of dissolved solid.)
 The type of substance obtained by this method is
heat-stable substance because it will not
decompose on strong heating. Eg. Sodium chloride
Solid - Solid

sulfur

Iron +
sulfur
 MAGNETIC
SEPARATION
 Make use of the special properties of the
substance
(mixture of iron filings and sulfur…use a magnet
a. Bring the magnet to the mixture.
b. The iron filings will be attracted to the magnet
while the sulfur will be left behind. )
 SUBLIMATIO
 is used when one of theNsolid sublimes
Eg.
a. mixture of iodine and copper
b. mixture of ammonium chloride + sodium chloride
 SIMPLE
DISTILLATION
 The principle of simple distillation is based on the
boiling point of a substance.
(A liquid boils and turns into vapor at its
boiling point. When the vapor is
condensed, the pure liquid is obtained again_
 The type of mixture to be separated by this method

is a mixture containing one type of liquid only.

Eg. seawater
 SIMPLE
DISTILLATION
• The mixture is heated in the distillation flask.
• The liquid with a lower boiling point in the
mixture boils and changes into vapour.
• The vapor flows into the condenser and is
cooled by cold water in it. The vapor
condenses and changes back to liquid.
• The liquid is collected in the receiving flask
which is called the distillate.
SIMPLE
DISTILLATION
SIMPLE
DISTILLATION
SIMPLE
DISTILLATION
 Fractional Distillation
 Is used to separate a mixture of miscible liquids
with different boiling points.
 Its principle is based on the different boiling points
of liquids.
 The liquid with the lowest boiling point is distilled
first. The liquid with the highest boiling point is
distilled last.
 The type of mixture to be separated by this
method is a mixture of two miscible liquids.
 Eg. Ethanol and water, petroleum, liquid air
Fractional Distillation
 Fractional Distillation
How is fractional distillation carried out?
• The mixture is heated until it boils.
• The liquid with the lowest boiling point boils first and changes into a
vapor. The temperature remains constant.
• The vapor enters the condenser and is cooled by the cold water. It
condenses back into a liquid.
• The liquid is collected in the receiving flask and is called the distillate.
• When the liquid of the lowest boiling point has been distilled over and
collected completely, the temperature in the mixture rises again.
• When the temperature reaches the next lowest boiling point of another
liquid, the second liquid is distilled and collected in a new receiving
flask.
(The process is repeated until all the liquids in the mixture are separated
into individual receiving flasks.)
Fractional Distillation
Fractional Distillation
Fractional Distillation
Fractional Distillation
Fractional Distillation
 Fractional Distillation
What are the industrial applications of
fractional distillation?

 To separate pure oxygen and nitrogen from

liquid air
 To separate crude oils into different fractions
 To separate ethanol from the fermentation of
sugars to make alcoholic beverages.
Fractional Distillation of Crude Oil
Separating Funnel
Separating Funnel
 Separating Funnel
How is the process carried out in using a separating
funnel?
• Close the tap and pour the mixture of oil and water
into the separating funnel with a stopper placed on
top. Let the immiscible liquids to settle out.
• Remove the stopper and place an empty beaker
below the tap. Open the tap slowly and carefully
drain the water out of the funnel into the beaker.
• When the water has completely drained out, close
the tap. In this way, water is separated from the oil.