METHODS OF PURIFICATION OF ORGANIC COMPOUNDS

Once an organic compound is extracted from a natural source or synthesised in the laboratory,
it is essential to purify it. Various methods used for the purification of organic compounds are
based on the nature of the compound and the impurity present in it.

The common techniques used for purification are as follows :

(i) Sublimation
(ii) Crystallisation
(iii) Distillation
(iv) Differential extraction and
(v) Chromatography

Finally, the purity of a compound is ascertained by determining its melting or boiling point.
Most of the pure compounds have sharp melting points and boiling points.

New methods of checking the purity of an organic compound are based on different types of
chromatographic (e.g Column Chromatography, Gas Chromatography; GC, High Performance
Liquid Chromatography; HPLC, etc) and spectroscopic techniques ( e.g Ultraviolet radiation;
UV, Infrared; Ir, Mass Spectroscopy; MS, X-ray Crystallography etc).

Sublimation

This method is used for solids that move directly from the solid state to the gas phase ( vapour
phase) without passing through the liquid phase. The purification technique based on the above
principle is known as sublimation and is used to separate sublimable compounds from
nonsublimable impurities.

Crystallisation

This is one of the most commonly used techniques for the purification of solid organic
compounds. It is based on the difference in the solubilities of the compound and the impurities
in a suitable solvent. The impure compound is dissolved in a solvent in which it is sparingly
soluble at room temperature but appreciably soluble at higher temperature. The solution is
concentrated to get a nearly saturated solution. On cooling the solution, pure compound
crystallises out and is removed by filtration. The filtrate (mother liquor) contains impurities and
small quantity of the compound. If the compound is highly soluble in one solvent and very little
soluble in another solvent, crystallisation can be satisfactorily carried out in a mixture of these
solvents. Impurities, which impart colour to the solution are removed by adsorbing over
activated charcoal. Repeated crystallisation becomes necessary for the purification of
compounds containing impurities of comparable solubilities.

Distillation
This important method is used to separate

(i) volatile liquids from nonvolatile impurities and

(ii) the liquids having sufficient difference in their boiling points.

Liquids having different boiling points vaporise at different temperatures. The vapours are
cooled and the liquids so formed are collected separately. Chloroform (b.p 334 K) and aniline
(b.p. 457 K) are easily separated by the technique of distillation.

The liquid mixture is taken in a round bottom (R.B) flask and heated carefully. On boiling, the
vapours of lower boiling component are formed first. The vapours are condensed by using a
condenser and the liquid is collected in a receiver. The vapours of higher boiling component
form later and the liquid can be collected separately.

Fractional Distillation: If the difference in boiling points of two liquids is not much, simple
distillation cannot be used to separate them. The vapours of such liquids are formed within the
same temperature range and are condensed simultaneously. The technique of fractional
distillation is used in such cases. In this technique, vapours of a liquid mixture are passed
through a fractionating column before condensation. The fractionating column is fitted over the
mouth of the round bottom flask as seen below.
Vapours of the liquid with higher boiling point condense before the vapours of the liquid with
lower boiling point. The vapours rising up in the fractionating column become richer in more
volatile component. By the time the vapours reach to the top of the fractionating column, these
are rich in the more volatile component.

Fractionating columns are available in various sizes and designs as shown below. A
fractionating column provides many surfaces for heat exchange between the ascending vapours
and the descending condensed liquid. Some of the condensing liquid in the fractionating
column obtains heat from the ascending vapours and revaporises. The vapours thus become
richer in low boiling component. The vapours of low boiling component ascend to the top of
the column. On reaching the top, the vapours become pure in low boiling component and pass
through the condenser and the pure liquid is collected in a receiver. After a series of successive
distillations, the remaining liquid in the distillation flask gets enriched in high boiling
component. Each successive condensation and vaporisation unit in the fractionating column is
called a theoretical plate. Commercially, columns with hundreds of plates are available.
Different types of fractionating columns.

One of the technological applications of fractional distillation is to separate different fractions of

crude oil in petroleum industry.

Distillation under reduced pressure: This method is used to purify liquids having very high
boiling points and those, which decompose at or below their boiling points. Such liquids are
made to boil at a temperature lower than their normal boiling points by reducing the pressure
on their surface. A liquid boils at a temperature at which its vapour pressure is equal to the
external pressure. Glycerol can be separated from spent-lye in soap industry by using this
technique. The pressure is reduced with the help of a water pump or vacuum pump as shown
below
Steam Distillation: This technique is applied to separate substances which are steam volatile
and are immiscible with water. In steam distillation, steam from a steam generator is passed
through a heated flask containing the liquid to be distilled. The mixture of steam and the
volatile organic compound is condensed and collected. The compound is later separated from
water using a separating funnel. In steam distillation, the liquid boils when the sum of vapour
pressures due to the organic liquid (p1) and that due to water (p2) becomes equal to the
atmospheric pressure (p), i.e. p =p1+ p2. Since p1 is lower than p, the organic liquid vaporises
at lower temperature than its boiling point. Thus, if one of the substances in the mixture is
water and the other, a water insoluble substance, then the mixture will boil close to but below,
373K. A mixture of water and the substance is obtained which can be separated by using a
separating funnel. Aniline is separated by this technique from aniline – water mixture.

Differential Extraction

When an organic compound is present in an aqueous medium, it is separated by shaking it with

an organic solvent in which it is more soluble than in water. The organic solvent and the
aqueous solution should be immiscible with each other so that they form two distinct layers
which can be separated by separatory funnel. The organic solvent is later removed by
distillation or by evaporation to get back the compound. Differential extraction is carried out in
a separatory funnel as shown
If the organic compound is less soluble in the organic solvent, a very large quantity of solvent
would be required to extract even a very small quantity of the compound. The technique of
continuous extraction is employed in such cases. In this technique same solvent is repeatedly
used for extraction of the compound.

Chromatography

Chromatography is an important technique extensively used to separate mixtures into their

components, purify compounds and also to test the purity of compounds. The name
chromatography is based on the Greek word chroma, for colour since the method was first used
for the separation of coloured substances found in plants. In this technique, the mixture of
substances is applied onto a stationary phase, which may be a solid or a liquid. A pure solvent,
a mixture of solvents, or a gas is allowed to move slowly over the stationary phase. The
components of the mixture get gradually separated from one another. The moving phase is
called the mobile phase. Based on the principle involved, chromatography is classified into
different categories.

Two of these are:

(a) Adsorption chromatography, and

(b) Partition chromatography.

Adsorption Chromatography: Adsorption chromatography is based on the fact that different

compounds are adsorbed on an adsorbent to different degrees. Commonly used adsorbents are
silica gel and alumina. When a mobile phase is allowed to move over a stationary phase
(adsorbent), the components of the mixture move by varying distances over the stationary
phase. Following are two main types of chromatographic techniques based on the principle of
differential adsorption.

(i) Column chromatography, and

(ii) Thin layer chromatography.
 Column Chromatography: Column chromatography involves separation of a mixture over
a column of adsorbent (stationary phase) packed in a glass tube. The column is fitted with a
stopcock at its lower end. The mixture adsorbed on soluble in the organic solvent, a very
large quantity of solvent would be required to extract even a very small quantity of the
compound. The technique of continuous extraction is employed in such cases. In this
technique same solvent is repeatedly used for extraction of the compound.

Thin Layer Chromatography: Thin layer chromatography (TLC) is another type of adsorption
chromatography, which involves separation of substances of a mixture over a thin layer of an
adsorbent coated on glass plate. A thin layer (about 0.2mm thick) of an adsorbent (silica gel or
alumina) is spread over a glass plate of suitable size. The plate is known as thin layer
chromatography plate or chromaplate. The solution of the mixture to be separated is applied as
a small spot about 2 cm above one end of the TLC plate. The glass plate is then placed in a
closed jar containing the eluant.

As the eluant rises up the plate, the components of the mixture move up along with the eluant
to different distances depending on their degree of adsorption and separation takes place. The
relative adsorption of each component of the mixture is expressed in terms of its retardation
factor i.e. Rf value

Distance moved by the substance from base line (x)

Rf =
Distance moved by the solvent from the base line (y)

The spots of coloured compounds are visible on TLC plate due to their original colour. The
spots of colourless compounds, which are invisible to the eye but fluoresce in ultraviolet light,
can be detected by putting the plate under ultraviolet light. Another detection technique is to
place the plate in a covered jar containing a few crystals of iodine. Spots of compounds, which
adsorb iodine, will show up as brown spots. Sometimes an appropriate reagent may also be
sprayed on the plate. For example, amino acids may be detected by spraying the plate with
ninhydrin solution.

Partition Chromatography: Partition chromatography is based on continuous differential

partitioning of components of a mixture between stationary and mobile phases. Paper
chromatography is a type of partition chromatography. In paper chromatography, a special
quality paper known as chromatography paper is used. Chromatography paper contains water
trapped in it, which acts as the stationary phase. A strip of chromatography paper spotted at the
base with the solution of the mixture is suspended in a suitable solvent or a mixture of solvents
This solvent acts as the mobile phase. The solvent rises up the paper by capillary action and
flows over the spot. The paper selectively retains different components according to their
differing partition in the two phases. The paper strip so developed is known as a
chromatogram. The spots of the separated coloured compounds are visible at different heights
from the position of initial spot on the chromatogram. The spots of the separated colourless
compounds may be observed either under ultraviolet light or by the use of an appropriate spray
reagent as discussed under thin layer chromatography.