ASSIGNMENT IN ORGANIC CHEMIS

TRY LAB
submitted by: evangelista, tristan nicole b. bsmt one b
submitted to: maam mabolo
PURIFICATION

What is
Purification in a chemical context is the physical separation of a chemical substance
Purification?

of interest from foreign or contaminating substances. Pure results of a successful

purification process are termed isolate.
The following list of chemical purification methods should not be considered
exhaustive.

Affinity
Purification

1. Affinity Purification is used to purify proteins by

retaining them on a column to their affinity to
antibodies, enzymes or receptors which have
been immobilized on the column.
Example:
Tandem

Affinity

Purification
(TAP)

is

purification
technique
studying

for
protein-

protein
interactions.
Principle:
Tandem
Affinity
Affinity Purification is one of the most diverse and
powerful
chromatographic methods
Purification
for purification of a specific molecule or a group of molecules from complex mixtures

 It is based on highly specific biological interactions between two molecules such as

interactions between enzyme and substrate, receptor and ligand, or antibody and
antigen.
These interactions which are typically reversible are used for purification by placing
one of the interacting molecules referred to as affinity ligand onto a solid matrix to
create a stationary phase while a target molecule is in the
Filtration
mobile phase.
solvent

Filtration is a mechanical method to separate solids

filter paper

from liquids

filter funnel

or gases by passing the feed stream

through a porous sheet such as a cloth or membrane,

residue

which retains the solids and allows the liquid to pass

through. The liquid which has passed through filter is

clamp

called the filtrate.

Example:
Using

filtrate

coffee

filter

to

separate

the

coffee

flavor from the coffee beans.

Principle:

Example of Filtration

The volume of filtrate collected per unit time (dV/dt) is

termed

as

the

rate

of filtration.
As the filtration proceeds, solid particle accumulate on the filter medium
forming

packed

bed

of

solids, called filter cake.

Centrifugation

3. Centrifugation is

process

in

which

light

particles are revolved at high speed with the help

of an electric motor so that the fine particles
which do not settle at bottom would settle down.
Example:

Centrifugation of

blood

separates

layers

into

Centrifugation
of
where
blood samples

particles

samples
the

(erythrocytes)
the

HEAVIEST
move

to

bottom
of the test tube.

Principle:
Centrifugation is based on the fact that any object moving in a circle at a steady
angular velocity is subjected to an outward directed force , F. The magnitude of
this force depends on the angular velocity in radians, omega, and the radius of
rotation, r, in cm.

F= 2r

Evaporation

4. Evaporation is used to remove volatile liquids from

non-volatile solutes which cannot be done through
filtration due to the small size of the substances.
Example:
Evaporation of water steam from kettle.
Principle:
The fastest moving molecules (those with
the highest kinetic energy) at the surface of the liquid have enough energy to
break the attractive bonds with other molecules. They then escape the
surface of the substance. Obviously, this only occurs with the molecules at
the surface of the substance.
Since at higher temperatures the molecules have more kinetic energy, more
of them are likely to escape, and so evaporation occurs more quickly at
higher temperatures.
In general, evaporation occurs because systems seek equilibrium (there is a
low concentration of molecules in the air, and a high concentration in the
liquid).

5. Crystallization separates a product from a liquid feed stream, often in

extremely pure form, by cooling the feed stream or adding precipitants which
lower the solubility of the desired product so that it forms crystals. The pure
solid crystals are then separated from the remaining liquor by filtration or
centrifugation.

Crystallization

Example: Crystallization of salt from sea water.

Principle:
Compounds (solutes) tend to be more soluble in hot liquids (solvents) than
they are in cold liquids. If a saturated hot solution is allowed to cool, the
solute is no longer soluble in the solvent and forms crystals of pure
compound.
6. Recrystallization: In analytical and synthetic chemistry work, purchased
reagents of doubtful purity may be recrystallized, e.g. dissolved in a very pure
solvent, and then crystallized, and the crystals recovered, in order to improve
and/or verify their purity.

Recrystallization

Example:
Recrystallization of fossils.
Principle:
The amount of solute that can be dissolved by a solvent increases with
temperature.

7. Distillation, widely used in petroleum refining and in

purification of ethanol separates volatile liquids on
the basis of their relative volatilities.
Example:
Distillation of water/ distilled
water.
Principle:

Distillation

Every element can exist in three states: as a liquid, as a solid

and as a vapor, which mostly depend on it's temperature. This
applies to water, too. So, water can be found as ice, water and
steam.

8. Water purification combines a number

of methods to produce potable
or drinking water.

Example:
Purified water.
Principle:
Works on the same principle as rapid gravity filters, differing in that the
filter medium is enclosed in a steel vessel and the water is forced through
it under pressure.

SEPARATION

What is
Separation?
A separation process,

or a separation technique, or simply a separation, is a

method
to
achieve
any mass
transfer phenomenon
that
converts
a mixture of substances into two or more distinct product mixtures (which may be
referred to as fractions), at least one of which is enriched in one or more of the
mixture's constituents.
The following are some methods of separating mixtures.

1. Precipitation is
any
product
of
the
condensation of atmospheric water vapour that
falls
under
gravity.
The
main
forms
of precipitation include drizzle, rain, sleet,
snow, graupel and hail.
Example:
Double diffusion (ouchterlony), Single diffusion
Immunoelectrophoresis and Immunofixation.

(radial

immunodiffusion),

Principle:

Soluble antigen + antibody (in proper proportions) ->

precipitate.
Lattice formation (antigen binds with fab sites of 2 antibodies).

visible

2. Sedimentation is
the
tendency
for
particles in suspension to settle out of the
fluid in which they are entrained and come
to rest against a barrier.
Example:
Grave
Sedimentation

l in water, sand in water.

Principle:
Heavy paricles settle faster in a
medium.

3. Decantation is a process for the separation of mixtures, by

removing a layer of liquid, generally one from which a
precipitate has settled.
Example:
Boiling tea bags in water to make the tea for iced tea. One has
to decant the brownish liquid from the tea bags, they remain
behind in the boiling pot and the tea is deposited into another
container.
Principle:
Separating a liquid from a solid by gently pouring the liquid
from the solid so as not to disturb the solid.

Decantation

4. Drying is a mass transfer process consisting of

the removal of water or another solvent by
evaporation from a solid.
Example:
Drying clothes by hanging it on a hanger facing
the sunlight.
Drying

Principle:

The migration of moisture from the interior of an individual grain to the

surface, and the evaporation of moisture from the surface to the
surrounding air.
The rate of drying is determined by the moisture content and the
temperature of the grain and the temperature, the (relative) humidity and
the velocity of the air in contact with the grain.

5. Chromatography is the separation of a mixture

by passing it in solution or suspension or as a
vapor (as in gas chromatography) through a
medium in which the components move at
different rates.
Example:
The separation of a mixture by passing it in
solution or suspension or as a vapor (as in gas
chromatography) through a medium in which the
components move at different rates.
Principle:

The basic paper chromatography system.

In chromatography a liquid is pumped through a bed of particles. The liquid is

called the mobile phase and the particles the stationary phase. A mixture of the
molecules that shall be separated is introduced into the mobile phase.
6. Flocculation is the process by which individual particles of clay aggregate into
clotlike masses or precipitate into small
lumps. Flocculation occurs as a result
of a chemical reaction between the clay
particles
and
another
substance,
usually salt water.
Example:
Flocculation of Algae inside the fish
tank.
Principle:
Flocculation processes constitute an important part of treatment of
surface water or of waters with high-level contamination.

7. Electrophoresis is a technique used in

laboratories
in
order
to
separate
macromolecules based on size. The
technique applies a negative charge so
proteins move towards a positive charge.
This is used for both DNA and RNA analysis.

Example:

Electrophoresis

The process of administering medicine, iontophoresis.

Principle:
An idealized, simplified situation: an isolated charged particle in a non conducting
medium. The force experienced by a particle in an electrical field is given by
Coulombs law, F = ZeE (E-electric field: potential per unit length) The viscous
resistance of the medium to the motion: -fv (f: the frictional factor) The viscous
resistance of the medium just balances the driving force. fv = F = ZeE.

8. Zone Refining is a method of purifying a crystalline

solid, typically a semiconductor or metal, by causing a
narrow molten zone to travel slowly along an otherwise
solid rod or bar to one end, at which impurities
become concentrated.
Example:
The preparation of a transistor
or diode semiconductor, an ingot of
germanium is first purified by zone
refining.
Then
a
small
amount
of antimony is placed in the molten zone,
which is passed through the pure germanium. With the proper
choice of rate of heating and other variables, the antimony can be
spread evenly through the germanium. This technique is also used
for the preparation of silicon for use in computer chips.
Principle:
Impurities are more soluble in the molten form than solid.