Methods of Purification and Analysis

Pure Substance – single substance not mixed with anything else. For example; white sugar, gold, iron,
copper sulfate crystals, distilled water etc
Impure Substance – contains two or more substances. Its quantity is more on Earth.
For example; seawater, milk (fats & dissolved solids), air, soil, steel etc

Obtaining Pure Substances

The components of mixtures do not lose their individual properties. They can be separated by using the
suitable physical method.
Purification – It the process of separation of mixtures into pure substances by using physical methods
without chemical reactions.
A physical property must be different for two or more substances to be separated.
It is important to determine the purity of substances in medicines, food and beverage industries etc.
The impurities in medicine, food and beverage can produce undesirable side effects. So purity is ensures
for safer consumption.

Filtration
Filtration – separates insoluble solid from a mixture usually liquid.
Filtration is used to separate a mixture of a liquid (or solution) and an insoluble solid. The insoluble solid is
collected as the residue while the liquid is collected as the filtrate.
Mixture is poured through a filter with tiny holes made of paper. Large solid particles cannot pass through
the pores and trapped in it as residue while tiny liquid particles pass through as filtrate.
For example; sand can be separated from a mixture of water and sand by filtration. The sand being
insoluble left over filter paper as residue and water is collected as filterate.

The purification of water requires different steps. The filtration is one the step in this process.

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Evaporation to Dryness
It is used for the separation of dissolved solid, which does not decomposed on heating to dryness, from a
solution. Examples: the group I metal carbonates and chlorides.

Crystallisation
It is used for separation of dissolved solid, which decomposed on heating to dryness from a solution.
Examples: Nitrates, sulfates, ammonium compounds etc

Why crystallisation occur?

The crystallization depends on the solubility of solute in a solvent.

Solubility: The amount of solute dissolved in 100g of solvent to make saturated solution at a given
temperature is called solubility of a solute.
Every solute has different solubility in a solvent at given temperature.
Solubility of most solutes decrease as temperature decrease. When solution cools, solution can’t hold more
solute (saturated) so the extra solute separates as pure crystals.

Process: Gently heat the solution to make saturated solution or to the crystallization point. Then gradually
cools it down. The solute separates as crystals. Filter out the crystals and wash with very little distilled
water and then dry in the folds of filter paper or sunlight.
For example; the crystals of copper (II) sulfate can be obtained from a saturated solution of copper (II)
sulfate by crystallization.

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Sublimation
It is used for separation of a mixture of solids in which one of solid sublimes from other solids with their
high melting point. Examples of solids that sublime on heating includes iodine, ammonium chloride,
naphthalene (moth balls) etc.
For example; iodine can be separated from a mixture of sand and iodine by sublimation.

When mixture of iodine and sand is heated, iodine sublimes (turns into vapour directly) then cools and
crystallise when it reaches colder area.

Separation of Immiscible Liquids (liquids which do not mix together)

The separating funnel is used to separate immiscible liquids on the basis of their different density. The
denser liquid will flow out first from the flask. For example; a mixture of oil and water.

The two liquids insoluble to each other will create two layers of overlying liquids of each type.
To separate the liquids, take the stopper off and turn the tap on to run the denser liquid at the bottom off
the funnel and leave the less dense liquid in the funnel by turning the tap off and reset the stopper at its
original position.

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Simple Distillation

It is used to obtain solvent from a solution containing non – volatile solute (high b.p.) and for separation of
mixture of liquids if the difference between their boiling points is more than 20 oC.
For example water from a solution of salty water.
Simple distillation consists of two processes in it.
• Evaporation
• Condensation

Process of Distillation:
Solution is heated in the distillation (boiling) flask. The steam (solvent) is produced which rises to the
condenser. The steam is cooled in the condenser to form liquid water. It is collected in the receiving flask.
Solute (salt) remains in the flask as it has high boiling point.

When the solution is heated, its temperature

increases. The thermometer records a temperature
of 100 oC when the solution boils.
This is the temperature of water vapours.
The temperature remains unchanged until all the
water has boiled off.

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Fractional Distillation

It is used to separates mixture of miscible (soluble) liquids on the basis of their different boiling points.
It also consists of two processes in it.
• Evaporation
• Condensation
A fractionating column is attached with the boiling flask and condenser. The glass beads in the
fractionating column provide the greater surface area for vapours to condense. In fractional distillation, the
liquid with the lowest boiling point distilled over first, then next and so on.
The vapours of liquids with higher boiling point condense in the fractionating column and return to the
boiling flask.

The fractionating column separates the mixture of liquids. The temperature in the fractionating column
decreases upward.
Thermometer measure the temperature of vapours reaching there. The temperature on thermometer
increases when first fraction distilled over and vapours of next fraction touch the bulb of thermometer.
Condenser will cool down the vapours and turn them into liquid state. The water must enter into
condenser from the bottom and leave the condenser from the top.
The mouth of receiving flask must be open. If it is closed then pressure builds up in the apparatus.

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The change in temperature as a solution of ethanol (78 oC) and water (100 oC) undergoes fractional
distillation is shown below.

The temperature of the mixture increases as it is

heated. The ethanol distilled over first at 78 oC.
The temperature remains constant until all the
ethanol distilled over. The temperature then
increase until 100 oC.
At this temperature, the water distills over.
The temperature remains constant as water is
being distilled.

Uses of fractional distillation:

• Separates pure oxygen and pure nitrogen from liquefied air
• Separates substances in petroleum (crude oil) into fractions
• Separates alcohol to produce alcoholic drinks

Chromatography

Chromatography – a method of separating and identifying mixtures.

There are different techniques of chromatography. The paper chromatography is one of them.

This technique used a solvent to separate the mixture into its components on the basis of their different
solubility in the solvent.

Procedure for Paper Chromatography

Take a strip of chromatographic paper.

Draw a line with a pencil across a piece of

chromatography paper. This line should be about
1 cm from the bottom of the paper. Do not use a
pen as the colours in the ink may move up the
chromatography paper with the solvent.
Put a spot, using a teat pipette or a capillary tube,
of the mixture of dyes on the pencil line and
allow it to dry.

Suspend the chromatography paper in a glass jar ot tank that contains a small amount of suitable solvent so
that the bottom of the paper goes into the solvent. It is important that the solvent is below the pencil line so
that the inks/colourings don't just dissolve in the solvent.

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Put a lid on the jar so that the atmosphere becomes saturated with the solvent. This is to stop evaporation
of the solvent from the surface of the paper.
When the solvent has moved up the paper to about 1 cm from the top, remove the paper from the jar and
draw a pencil line to show where the solvent got to. The highest level of the solvent on the paper at any
time is called the solvent front.
Leave the paper to dry so that all the solvent evaporates.
The different solute in the sample separates on the basis of their different solubility in the solvent.

The dye will dissolve in solvent and travel up the

paper at different speed. Hence they are separated
and coloured spot appeared in different places on
the paper. The dye (solute) which is more soluble
in the solvent will travel a longer distance from
the starting line and vice versa. The patter of dyes
(solute) obtained on the paper is called a
chromatogram.

The colouless substances are also separated and analysed by chromatography. After the same
chromatographic process as mentioned above, the locating agent is sprayed onto the paper.
The locating agent reacts with the colourless substances and makes the coloured spots. Thus their position
becomes visible.

Rf Value
Rf stands for retardation factor. It is defined as; It is the ratio between the distance travelled by the
solute (dye) and the distance travelled by the solvent front is a constant. This ratio is called the Rf value of
the substance.

distance travelled by solute

Rf =
distance travelled by solvent front

Rf is the ratio thus has no unit. The value of Rf

does not change as long as chromatography is
carried out under the same conditions i.e. same
solvent and same temperature. The Rf value of a
substance is less than one (1) but more than Zero
(0). If a substance has zero value of Rf then it is
insoluble in the solvent. Bigger the value of Rf
then more is the solubility of substance in that
solvent and vice versa.
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Using Paper Chromatography in Analysis
The paper chromatography can be used to identify the particular dyes in a mixture. If you think that your
mixture (m) could contain dyes d1, d2, d3 and d4 then you can carry out an experiment to determine this.
A pencil line is drawn on a larger sheet of paper and pencil marks are drawn along the line to show the
original positions of the various dyes placed on the line. One spot is your unknown mixture; the others are
single, known dyes. The chromatogram is then allowed to develop as before.

The mixture (m) has spots corresponding to dyes

d1, d3 and d4. They have the same colour as
spots in the mixture, and have travelled the same
distance on the paper. Although dye d2 is the
same colour as one of the spots in the mixture, it
has travelled a different distance and so must be a
different compound.

Paper Chromatography can be used to;

• Separates dyes in the ink and pigment in the plants
• Separates amino acids obtained from proteins
• Identify poison or drugs and to detect the traces banned substances in food

Checking the Purity of Substances

Many substances are not pure in the nature. It is important to determine the purity of the substances.
For example; impurity in medicine must be detected as impurity may produce undesirable effects when the
medicine is consumed.
Purity is also important in the food and beverage industry. Chemicals such as preservatives and dyes used
in food and beverages must be safe for consumption.
Melting point, boiling point and chromatography are used to check the purity of substances.

Melting Point
Every pure substance has a fixed value of melting point. The temperature remains constant in melting
process. There are two effects if the impurity is present in a substance.
• Melting point of a substance decreases
• Substance melts over a range of temperature
Melting point is usually used to check the purity of solids.

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Boiling Point
Every pure substance has a fixed value of boiling point. The temperature remains constant in boiling
process. There are two effects if the impurity is present in a substance.
• Boiling point of a substance increases
• Substance boils over a range of temperature
Boiling point is usually used to check the purity of liquids.

Chromatography
There are certain substances which decomposed before the melting and boiling points.
Chromatography can be used to check the purity of such substances. A pure substance will produce only one
well-defined spot on a chromatogram. If impurities are present then several spots will be seen on the
chromatogram.

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