ISOLATION PURIFICATION AND

STRUCTURAL DETERMINATION OF
ORGANIC COMPOUNDS

CHM 11
LECTU 2
RER : M
PERIOD R S BAKE
S : 10-1 R M.T
2noon
Monda o n
ys
 INTRODUCTION
ORGANIC COMPOUNDS WHETHER ISOLATED FROM NATURAL SOURCES OR SYNTHESIZED IN THE LABORATORY NEEDS TO BE PURIFIED.
THIS IS INORDER TO OBTAIN COMPOUNDS THAT ARE OF HIGH PURITY TO BE UTILIZED FOR VARIOUS PRUPOSES SUCH AS
PHARMEACEUTICALS, NEUTRACEUTICALS, COSMETICS, BIOMEDICS, E.T.C. SEVERAL METHODS CAN BE EMPLOYED IN THE SEPERATION
AND OR PRIFICATION OF THESE ORGANIC COMPOUNDS DEPENDING ON THE NATURE OF THE MIXTURE:

• CRYSTALLIZATION

❑ SUBLIMATION

❑ SIMPLE DISTILLATION

❑ CHROMATOGRAPHY

❑ VACUUM DISTILLATION

❑ STEAM DISTILLATION

❑ FRACTIONAL DISTILLATION

❑ DIFFERENTIAL EXTRACTION
 CRYSTALLIZATION
The principle of crystallization is based on the difference in the solubilities of the
compound and the impurities in a suitable solvent.

The impure compound is dissolved in the 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 the filtration.

The filtrate contains impurities and small quantity of the compound. If the
compound is highly soluble in one solvent and very little soluble in another solvent ,
then, crystallisation can be carried out.

Impurities which impart colour to the solution are removed by adsorbing over
activated charcoal.

For example, impure benzoic acid can be purified by dissolving in hot water, an
then adding activated charcoal to adsorb the impurities, filtering the solution of
benzoic acid and activated charcoal and then crytallizing the solution by leaving
the solution to cool down. This is because, benzoic acid is insoluble in cold water
but soluble in hot water.
SUBLIMATION

Sublimation is the transition that substances undergo when they change

from solid directly to the gaseous state without undergoing the liquid phase.

✔ It is used to separate sublimable compound from non-sublimable

impurities.

✔ In this process, the substance to be sublimed heated in a crucible which

has an inverted funnel kept above it to collect the sublimable compounds.

✔ To hasten the process, the funnel is kept cool.

✔ An example of sublimation process is the formation of fog which in

involves the changing of dry ice (solid form of CO2) directly from solid
state to the gaseous state. OtherExamples of solid that sublime include:
solid water (ice), arsenic and iodine. Sublimation is used as a purification
process for volatile organic compounds.
SIMPLE DISTILLATION

This purification process is used in the purification of miscible liquids or solvents with large
difference (about 25⁰C) in boiling points. Boiling point of a liquid is the point at which the vapour
pressure of a liquid is equal to its atmospheric pressure. Simple distillation is used to separate; (i)
Volatile liquids from nonvolatile impurities (ii) liquids having sufficient difference in their boiling
points because, liquids having different boiling points vaporize at different temperatures.

In this method, the vapours are cooled and the liquids so formed are collected separately.
Chloroform (B.P= 60⁰C) and aniline (BP.= 189⁰C) are easily separated by the technique of distillation
. The liquid mixture is taken in a round bottom 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.
VACUUM DISTILLATION OR 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.

Therefore, since the boiling point is dependent on the atmospheric pressure, the liquids will
boil at a temperature lesser than their boiling points if they were distilled in an atmosphere
having lower pressure.

The pressure is reduced with the help of a water pump or vacuum pump therefore, making the
distillation faster and preventing the liquid that decomposes close to or at its boiling point from
doing so . Glycerol can be separated from spent-lye in soap industry by using this technique.
VACUUM DISTILLATION SET UP
STEAM DISTILLATION
This technique is applied to separate substances which are steam volatile and are immiscible with
water. 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 vaporizes 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 .

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.

STEAM DISTILLATION SET-UP
FRACTIONAL DISTILLATION
Fractional distillation involves the separation of two miscible organic liquids/ solvent
having little difference in boiling points. 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 • 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.
FRACTIONAL DISTILLATION
• 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 used.

FRACTIONAL DISTILLATION
DIFFERENTIAL EXTRACTION
This method is used for immiscible liquids, that is, liquids that do not mix together. For example, oil
and water are immiscible.

The immiscible liquids are taken in a separating funnel and left undisturbed. After a while, they
separate out according to their specific gravities, with the heavier liquid at the bottom. Then they
are later collected.

Substances can also be separated according to their preferential solubility in the liquid. For
example, if phenol is to be extracted, it can be preferentially extracted using NaOH solution as one
of the liquids used.
 CHROMATOGRAPHY
Chromatography is a laboratory technique used for the separation and purification of a mixture of organic compounds.
The mixture is dissolved in a fluid called the mobile phase, which carries it through a system on which is fixed a material
called the stationary phase. The mobile phase is allowed to move slowly on the stationary phase. Due to which the
components of the mixture start separating from one another.

Chromatography is of two types:

Adsorption Chromatography

Partition Chromatography

Adsorption Chromatography

It is based on the principle that the constituents are adsorbed on an adsorbent in varying degrees. The adsorbents used
are generally silica gel or alumina. When a mobile phase moves over the fixed phase, different constituents of the mixture
get adsorbed at various distances over the fixed phase.

Adsorption Chromatography is further classified into:

Column Chromatography
CHROMATOGRAPHY
Column Chromatography

Here, a mixture is separated over a column of either silica gel or alumina, packed in a glass column. The constituent with the most affinity with the fixed phase is
adsorbed at the top, and so on. It is then retrieved by using an eluent. The solvent is then evaporated to get the constituent.
CHROMATOGRAPHY
Thin Layer Chromatography

Here a sheet of alumina is taken ( 0.2 mm thick) over which a small spot of the mixture is placed and it is kept in a suitable solvent. The solvent rises due to capillary
action and the constituents also rise with the solvent depending on their differential adsorption, and thereby, they are separated.
CHROMATOGRAPHY
Partition Chromatography: This is a process of separating mixtures of chemical compounds by passing them through a
column that contains a solid stationary phase that was eluted with a mobile phase.
This technique can be used to separate mixtures of closely related chemicals such as amino acids for identification and
further study.

Partition Chromatography is further classified into:

Paper Chromatography

Gas-Liquid Chromatography

Liquid-Liquid chromatography
CHARACTERIZATION (STRUCTURAL DETERMINATION) OF ORGANIC
COMPOUNDS
Characterization of organic compounds involves the determination of structures of organic
compounds using modern Instrumental techniques. The most common methods used are the
Spectroscopic and spectrometric methods.

Spectroscopic methods includes:

INFRARED SPECTROSCOPY (IR)

NUCLEAR MAGNETIC RESONANCE SPECTROSCOPY (NMR)

UV/VISIBLE SPECTROSCOPY (UV/VIS)

Spectrometric Methods includes:

MASS SPECTROMETRY (MS)

CHARACTERIZATION (STRUCTURAL DETERMINATION) OF ORGANIC
COMPOUNDS
Infrared Spectroscopy shows the functional groups present in an organic compound

Nuclear Magnetic Resonance establishes the connectivity of carbons and Hydrogen in a

compound. It is subdivided into: Proton (1H) NMR and Carbon-13 (13C) NMR

Mass spectrometry basically determines the Molecular mass of an organic compound

UV/Vis shows the wavelength of absorption of some functional group in the ultra violet (for
non-coloured compounds) and visible (for coloured compounds) regions of the electromagnetic
spectrum