CENTRIFUGATION

➢ Centrifugation is a process of phase separation by the application of centrifugal

force.
➢ It is possible, when the separation of phases in difficult by normal filtration.

CENTRIFUGE (Equipment):
It is a Rotating machine, in which centrifugal force is applied.
USES:
Separation of solid from liquid (or) separation of two immiscible liquids.

OBJECTIVES:
1. To obtain almost, dry solid cake from suspension.
2. To separate two immiscible liquids based on density.
3. To obtain insulin in pure state.
4. To evaluate the sedimentation rate in suspension and shelf-life of emulsion.
5. To determine the molecular weight of colloidal compounds.
6. To isolate bacterial cells, fungal cells from fermentation media.
7. To separate crystals such as aspirin from mother liquor.
8. To receiver lanolin from wool.
9. To separate drugs from body, urine, etc,..
10. To separate organic, inorganic, biological (or) microbiological, crystallin
compounds from suspension.
11. To refine/purify oils of animals and vegetable origins.
eg: Olive oil, fish liver oil etc,.
PRINCIPLE:
CRITICAL SPEED:
Speed at which the frequency of rotation matches the natural frequency
rotating part and the vibration.
CENTRIPETAL FORCE:
It is a force that acts towards the centre, hence it brings the object towards
the centre of rotation.
CENFRIFUGAL FORCE:
It is a force that acts against the centripetal force with an equal and opposite
rotation. Hence, it brings the object away from the centre of rotation.
CONDITIONS:
1. STATIONARY BOWL:
Influence of gravity, solids settle down liquids floats above the solid.
2. ROTATING IMPERFORATED BOWL:
Bowl is rotated about its vertical axis.
Both solid and liquid acted upon by gravity act as downward direction and
force act as horizontally.
3. ROTATING PERFORATED BOWL:
Bowl rotated about its vertical axis also perforated.
Influence of centrifugal force, solids tightly pressed against the inner
vertical wall of the bowl.
Shelf stress: directly proportional to square of the diameter of the centrifuge.
➢ An empty centrifuge is rotated, it create a stress on the centrifuge wall.
➢ At minimum self stress in smaller diameter centrifuge, a high centrifugal force is
achieved.

PRECAUTIONS:
➢ Material of construction have at least 1% elongation properties. So that risk of cracking
is decreased.
eg: Carbon steel (or) stainless steel with rubber (or) neoprene coating.
➢ For safe operation, damping is provided by elastic cushions.
APPLICATION OF CENTRIFUGATION:
1. In production of biologicals. eg: Insulin.
2. In biopharmaceutical analysis. eg: separation of drug from blood, urine
etc,..
3. In bulk drug industry. eg: crystals of aspirin.
4. Standardization of biphasic system.
5. Determination of molecular weight of colloids.
6. Isolation of bacterial cells, mycelium etc,.,
7. In refining (or) purification of animals (or) vegetable oil
8. In paint and varnish industries.
PRINCIPLE OF CENTRIFUGATION (OR) THEORY:
Principle of sedimentation, where the acceleration at centrifugal force causes
denser substance to separate out along the radical direction at the bottom of the tube.
➢ Lighter objects will tend to move to the top of the tube.
➢ Higher dense object will sink (sediment).
➢ If there is no different in density (isopycnic conditions), the particles stay steady.
➢ Even tiny difference in density to separate particles, gravity can replaced by more powerful
centrifugal force.
➢ Size about 5µm sediment at bottom, separated by simple filtration.
➢ Less than 5µm undergoes Brownian motion, a stronger centrifugal force is applied to
separate.
 V F = mv2
M R
r F F = Centrifugal force
center of centrifuge centrifuge tube V = Velocity of the body
R = Radius of circle of rotation

Gravitational force acting upon the same body

G = mg Acceleration due to gravity
Centrifugal effect is the ratio of the centrifugal force

Centrifugal force = Gravitational force acting radially

Gravitational force
C = F/ G
= mv2/ mgr
= v2/gr
V = 2π rn

n speed of rotation, unit s-1

 F/G = (2π rn)2
Gr
= 4π2 r2 n2
Gr
= 4π2 r2d2
G
Centrifugal force directly proportional to diameter, square of speed of rotation with same six
of centrifuge at high speed than large centrifuge.
TYPES OF CENTRIFUGAL SEPARATORS:
Two types of centrifugal techniques,
1. Differential centrifugation (or) differential pelleting.
2. Density gradient centrifugation
i) Rate zonal centrifugation
ii) Isopycnic centrifugation.
1. DIFFERENTIAL CENTRIFUGATION:
• Particles of different densities (or) sizes in a suspension will sediment at different
rate with larger and denser particles sedimenting faster.
• Sedimentation rate increased by centrifugal force.
• Suspension of cells subjected to a series of increasing centrifugal force will yield a series of
pellets with decreasing sedimentation rate.

A B C D
USES:
Harvesting cells (or) producing crude subcellular fractions from tissue
homogenate.
DISADVANTAGE:
Contamination.
2. DENSITY GRADIENT CENTRIFUGATION:
➢ Preferable for purification of subcellular organelles and macromolecules.
➢ Placing larger ( top) after layers of ( down) gradient media as sucrose in tube.
➢ Layer placed in discontinuous mode.
➢ In top place cell fraction, then centrifuged.
i. RATE- ZONAL CENTRIFUGATION:
• Cross-contamination avoided by layering the sample as a narrow zone on top of a
density gradient.
• Gradient stabilizes the bands and provides a medium of increasing density and viscosity.

Sample
zone
C.F Density
gradient
• Speed at which particles sediment depends their size and mass instead of density.
• Band more down through the density medium.
• Particles of similar sizes form as faster sedimenting move ahead of the slower once.
• Because of the density of the particles is greater than the density of the gradient all
particles will eventually form a pellet.

2. ISOPYCNIC CENTRIFUGATION (or) BUOYANT (or) EQUILIBRIUM SEPERATION:

• Particles separated on the basis of their density.
• Particle size only affects the rate at which particles move until their density is the same as
the gradient medium (density must be greater than density of particles).
• In this method, particles will never sediment to bottom of tube.

uniform mixture under centrifugal force particle

with gradient move until their density is same
medium to medium
• Particles of a specific density sediment until reach the point, where their density in
the same to media (i,e) equilibrium position.
• The gradient is then solid to isopycnic and particles separated according to
buoyancy.
• Density of biological particles is sensitive to osmatic pressure of gradient so its
depend on gradient medium used.
USES:
• Analytical purpose.
• Separation of biological.
eg: lymphocytes.
CENTRIFUGES:
MAJOR COMPONENTS:
DRIVE MECHANISM:
It is a source of rotary motion and it powered by an electric motor air, oil,
turbines etc,..
ROTOR:
It is the large rotating element of a centrifuge into (or) onto which sample is
loaded. It is drive fixed axis (or shaft) by drive mechanism with energy.
It should well-balanced, so as to decrease vibration and strain on tha shaft.
TYPES:
Low speed to increase of ~ 5×103 rpm
Based on temperature High speed to increase of ~ 2×104 rpm
Ultracentrifuge to maximum of ~ 105 rpm

Centrifuge in cold condition 1-40 c.

TYPES OF ROTOR:
1. Fixed angle rotor
2. Swinging bucket rotor.
1. FIXED ANGLE ROTOR:
Sample placed into shields (or) opening in the rotor at are particular pre-set
angle then tilled with their tops closer to shaft than their bottom.
2. SWINGING BUCKET ROTOR:
• Tubes are initially vertical.
• Bottom of sample tube then swing outward as shaft rotates.
• Tubes actually run horizontally.
• Useful for density gradient centrifuge.
ADVANTAGE:
Shows sedimentation as parallel bands.
CLASSIFICATION:
1. BASED ON MECHANISM:
Centrifuge

centrifugal sedimentors centrifugal filter

eg: non-perforated basket centrifuge eg: perforated basket centrifuge
2. LABORATORY/SMALL SCALE:
a) Swing-out type
b) Angle type
c) Ultra centrifuge
3. COMMERCIAL/ INDUSTRIAL LARGE SCALE:

Perforated bowl type/filtration Non perforated bowl type/

sedimentation centrifuge

Batch operation semi-continuous continuous Batch operation semi-continuous continuous

eg:1. top driven eg: Perforated eg: 1. top driven eg: conical disk
2. under driven semi-continuous/ 2. under driven centrifuge
centrifuge Short cycle/
Automatic batch
centrifuge Horizontal verticle
centrifuge centrifuge
CENTRIFUGAL FILTERS:
PRINCIPLE:
In centrifuge filter solid phase gets retained on the porous medium due to
difference in the densities of two phases ( solid and liquid).
Centrifugal filter consist of a porous wall (filter medium) which retain the
solid phase to pass through it.
Followings example of centrifugal filters.
1. Perforated basket centrifuge
2. Perforated semi-continuous centrifuge.
MERITS:
1. Suspension of high solid content can be centrifuged.
2. The final product can be obtained with very low moisture content.
3. Separation of the dissolved particle from the solid cake is possible.
DEMERITS:
1. Continuous operation is not possible.
2. Removal of hard cake is difficult with is formed due to centrifugal force.
3. Its operation is complicated.
4. Non economical.
APPLICATION:
1. If the solid phase recovery is the primary aspect, the centrifugation is the more
appropriate separation technique.
2. In the removal of undesirable solid particles from liquid phase.
eg: Removal of precipitated protein in the process of insulin manufacture.

PERFORATED BASKET CENTRIFUGE:

PRINCIPLE:
This centrifuge is based on the filtration mechanism, in which solid can be separated into
the porous medium due to difference in densities of solid and liquid phase. The basket
contain a porous wall which is covered with a filter medium. During centrifugation liquid
phase passes through a porous medium while solid particles are retained in the basket.
CONSTRUCTION:

1. It contains a perforated basket which may be made of any suitable metal like steel
(or steel coated with vulcanite) copper or metal.
2. These material should be corrosion resistant.
3. The diameter of basket is 0.9m and its capacity is 0.085m3 .
4. Perforation should be selected based on the size of solid to be filtered.
5. If the size of perforation is bigger than the particle to be filtered, then filter cloth is
used as lining inside the basket.
6. The basket is mounted on a vertical shaft, rotated by a motor with the help of
suitable power system.
7. The starting power of motor is 5kw and running power is 2kw.
8. A casing around the perforated basket is provided to collect the filtrate and to
convey it to outlet.
WORKING:
1. Feed is introduced into the basket while it is stationary. Feed should be optimum so
as to minimize the stream on the basket while centrifugation.
2. Feed should be loaded in the basket in proper manner.
3. The basket is operated at 1000 rpm.
4. It requires high power for starting i,e 5kw speed, but the power required for running
is small i,e 2kw.
5. Due to centrifugation, the solid particle get retained on the porous medium, while
the liquid passes through the membrane and is collected at the discharge.
6. The cake that is formed is made to dry by spinning it within the basket.
6. When the drying is complete the motor is switched off.
7. The cake can be separated by cutting it with a blade which is then manually
collected and unloaded.
MERITS:
1. Very compact and occupies less space.
2. It can separate solid from slurry containing high proportion of solid.
3. Highly viscous liquid can also be centrifuged.
4. The product obtained very low moisture content compared to product of filtration.
5. It separate the dissolved solid while in thermal drying process, the dissolved solid
cannot be separated.
6. The centrifugal process of rapid.
DEMERITS:
1. The operation is complicated.
2. It is a batch operation, so operation is not possible.
3. The centrifuge cannot be used for continuous operation because it undergo large amount
of stress.
4. Forms a hard cake whose removal is difficult.
APPLICATION:
1. Separation of drug crystals from mother liquor.
eg: Aspirin
2. Removal of undesirable solid from liquid.
eg: Separation of precipitated protein from insulin.
3. Centrifuge is useful for separating and drying crystalline solid which result in free-flowing
product.
PERFORATED SEMI-CONTINUOUS CENTRIFUGE:
PRINCIPLE:
Perforated semi-continuous centrifuge also known as short cycle or automatic batch
centrifuge works on the principle of filtration. The separation occurs through a filter medium
based on the difference in density of solid and liquid phase.

CONSTRUCTION:
1. It consist of a vertical perforated basket which rotates on a horizontal axis.
2. The slurry to be filtered is introduced from the feed tube.
3. Wash pipe (for washing the filter cake) is also introduced parallel to the feed pipe.
3. The feeds controls the thickness of feed and automatically stop its supply.
4. When the discharge of crystal is desired the hydraulic cylinder is attached in such
as way that the discharge chute inter the basket from its sides.
WORKING:
1. Feed is introduced in the basket through the feed tube.
2. The basket is allowed to rotate along the horizontal axis
3. Due to the process of centrifugation, solid gets retained in the filter in a basket and the
filtrate have the basket from its outlet the solid crystals are washed and the washed
water is drained out which from the outlet
4. The feeder maintain the feed thickness i,e a thickness of 50-70mm is preferred.
5. When the hydraulic cylinder is attached along with the discharge chute. The knife is
lifted, it cuts the filter cake such that a layer of crystal is left back. This layer of crystal
now serve a filter medium in next cycle.
6. The feeder and the hydraulic cylinder makes the process semi-automatic, hence the
name semi-automatic/semi-continuous centrifuge.
7. The crystal obtained contain 2-4% of moisture.
MERITS:
Semi-continuous centrifuge is semi-automatic.
DEMERITS:
1. Crystal may brake while discharging.
2. Designing and operation of the centrifuge is complicated.
3. Increased labour requirements.
4. Higher power consumption.
CENTRIFUGAL SEDIMENTATION:
Commonly employed centrifuged sedimentors include, continuous horizontal centrifuge,
vertical centrifuge, conical basket centrifuge/De-Laval clarifier and non-perforated basket
centrifuge.
APPLICATION:
1. Removal of precipitated protein from insulin liquor in insulin manufacturing.
2. Purification of oils (olive and fish liver oil) by removing dirt and water.
3. Separation of plasma from whole blood.
4. Collection of starch after washing and purification.
5. In purification of bacterial enzymes.
eg: Manufacturing if antibiotics.
6. Rapid separation of suspension having low solid concentration.
7. Recovery of lanolin from wool by scouring process.
MERITS:
1. These centrifuge occupy very less space.
2. They are highly effect in case of sedimentation of a sludge
3. Separation of coarse particle from fine particle is possible by controlling the rotation speed
and flow rate.
4. Separation of immiscible liquid can be achieved on continuous operation.
DEMERITS:
1. Construction is complicated.
2. Limited capacity.
NON-PERFORATED BASKET CENTRIFUGE:
PRINCIPLE:
Non-perforated basket centrifuge works on the principle of sedimentation. It cause
sedimentation of solid particle based on the difference in the densities of solid and liquid phase.
DESING:

1. A non-perforated basket is suspended on a vertical shaft.

2. It is made of suitable metal like steel.
3. The basket is driven by a motor.
4. Skimming tube is used for removing low density solid which gets deposited at liquid
surface.
WORKING:
1. It is an underdriven non-perforated basket centrifuge.
2. The suspension/slurry is introduced into the basket through the feed inlet.
3. The motor is started to rotate the basket at high speed.
4. The high density solid get deposited at the bottom and on the side walls of the basket, while
the liquid floats on the top.
5. When a considerable amount of solid get deposited, the motor is switched off. The liquid is
separated and then the solid is separated off manually.
6. The low density solid get deposited on the surface of the liquid. Hence, a skimming tube is
used for the separation of solid.
APPLICATION:
This centrifuge is useful only when the solid offer high resistance to liquid so that the centrifuge
process becomes easy.
SUPERCENTRIFUGE- VERTICLE CENTRIFUGE:
PRINCIPLE:
It works on the principle of sedimentation. It enables the separation of two immiscible
liquid based on the difference in their densities due to the centrifugal force, the heavier liquid
is thrown against the wall. While the lighter liquid from the inner layer (near center).
DESIGN:
1. It consist of a hallow cylinder bowl.
2. The diameter ranges from 1.75-5 inches with a length 9-30 inches.
3. It is attached to a flexible spindle at the top and to a loose-fit bushing at the bottom.
4. The bowl is mounted in such as way that it can be rotated on its longitudinal axis.
5. The liquid which is is to be separated is introduced into the bowl under pressure through
feed inlet.
6. Two outlet provisions are made at different height at top of the centrifuge.
7. Modified weirs can be used for simultaneously recovery of separated liquid.
WORKING:
1. Feed is admitted the bottom of the centrifuge through a feed nozzle under vaccum system.
2. Power is supplied to the entire system to rotate it in vertical manner.
3. The speed of rotation is up to 2000rpm.
4. By the centrifugation process, the heavier liquid phase is thrown out against the wall, while
the lighter liquid settle as an inner layer.
5. Hydraulic cylinder maintains, the liquid-liquid interface (neutral zone) between two
immiscible liquid.
6. Separated liquid phase are collected from the top of the bowl through outlet with the help
of modified weirs.
7. Simultaneous removal of two immiscible liquid is possible enabling continuous operation.
MERITS:
Efficiency of separation is high.
DEMERITS:
Output is limited.
Modification:
Tubular bowl centrifuge:
1. Used for the separation of solid and liquid when solid content is low.
2. From a suspension, the solid get accumulated on the wall, while the liquid can be clarified
from the top of centrifugal force.
3. If necessary, the deposited solid is collected at regular intervals.
4. Limited capacity.
APPLICATION:
1. These are widely employed in food, biochemical and pharmaceutical industry.
2. Used to separate phase of emulsion.
3. Clarification of liquid can also be carried out.