Topic 2

Separation and Purification

Importance of purification
It

is important that substances used in the food,

pharmaceutical and chemical industries are pure.
Test

Pure substance

Impure substance

Melting point of a
solid

Sharp/definite
temperature

Melts over a temperature

range.
T pure > T impure

Boiling point of a
liquid
(the impurity is a
dissolved solid)

Sharp/definite
temperature

Boils over a temperature

range
T pure < T impure

Boiling point of a
liquid
(the impurity is
another liquid)

Sharp/definite
temperature

Boils over a temperature

range - starts to boil at the
b.p. of one liquid and rises to
the b.p. of the other

Chromatography

Usually produces
only 1 spot on
chromatogram

Produces more than one spot

Separating and purifying substances

To

make sense of the material world around us, we

need the methods for physically separating the
many and varied mixtures that we come across.

The

most useful separation method for a particular

mixture depends on:
1. the type of mixture, and
2. which substance in the mixture we are most
interested in

Mixture

Method

Example

use some difference in

properties (density,
solubility, sublimation,
magnetism)

iron powder from

sand

suspension of solid
(insoluble) in liquid

filtration or centrifugation

sand from sea water

two/more of
immiscible liquids

Decantation or separating
funnel

water and oil

solution of solid in
liquid

to obtain solid
evaporation/crystallisation

salt from sea water

solid + solid
(powdered
mixture)

to obtain liquid distillation

two/more liquids
mixed together
(miscible)
solution of
two/more solids
(soluble) in a liquid

pure water from

aqueous copper
sulphate

fractional distillation (based

on differences in boiling
point)

to separate different
petroleum fractions

chromatography

a mixture of food
colouring in food
sample

Separating mixture of solids

Separating insoluble solids from liquids -1-

Filtration

Separating insoluble solids from liquids -2-

Decantation

Separating immiscible liquids

Separating solution of solid in liquid to obtain solid -1-

Crystallisation

Separating solution of solid in liquid to obtain solid -2-

Evaporation to dryness

Separating solution of solid in liquid to obtain liquid

Distillation

Separating two/more miscible liquids

Fractional Distillation

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,
determine the purity of a sample.

Chromatography

is a technique of using a solvent to

separate a mixture into its components.

Paper

chromatography is probably the simplest

form to set up and is very useful if we want to
analyse the substances present in a solution.

Paper chromatography

Chromatography paper
Pencil line
Mixture to be separated
Solvent e.g. ethanol

How does chromatography work?

For example, we need to determine

the purity of a food colouring (which

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

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 colouring is a mixture of two
dyes. It is not pure.

How does the chromatogram of a pure

substance look like?

However, if there is only one spot

on the chromatogram, it means the
substance is pure.

What are Rf values? -1The positions of the solvent front and spot on a
chromatogram depend on how long the experiment was
allowed to run.

Chromatogram
after a period of

Chromatogram
after a longer

What are Rf values? -2The ratio between the

distance travelled by
the substance
and
the distance travelled
by the solvent is a
constant

This ratio is called the Rf value of the

substance.
Rf value = distance moved by the

Examples notice the same Rf values

Rf = 3 cm / 4.5 cm
= 0.67

Chromatogram
after a period of

Rf = 3.5 cm / 5.2 cm
= 0.67

Chromatogram
after a longer

Rf values
The Rf value of a substance does not change as
long as chromatography is carried out under the
same conditions
(i.e. same solvent and same temperature).
This property allows us to
substance on a chromatogram.

easily

identify

How can we identify a banned substance present in

food colouring?

Chromatography

was performed on a sample of food

colouring (X) and 4 banned dyes ( A, B, C and D).

If

X contains any of the 4 banned dyes, it is not safe to

be consumed.

What conclusions can be drawn from this

chromatogram?

- 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.

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.

How do we identify colorless substances?

Chromatography can also be used for colorless
substances such as amino acids.
To separate and analyse colorless substances, we
apply a locating agent on a chromatogram.

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