Syllabus

Centrifugation:

Objectives, principle & applications of Centrifugation, principles, construction, working,

uses, merits and demerits of Perforated basket centrifuge, Non-perforated basket centrifuge,
semi continuous centrifuge & super centrifuge.

Chapter Outline

1) What is centrifugal force and centrifugation? Applications of centrifugation

2) Principles, construction, working, uses, merits and demerits of
a) Perforated basket centrifuge,
b) Non-perforated basket centrifuge,
c) semi continuous centrifuge
d) & super centrifuge.

 What is centrifugal force and centrifugation? Applications of centrifugation

Q.1 What is centrifugal force and centrifugation?

Ans- The earth's gravitational force is sufficient to separate many types of particles over time.
However, the length of time required in this manner of separation for most applications is
more and therefore in practice, centrifugal force is necessary to separate most particles.

Particles having a size above 5 micron sediment at the bottom due to gravitation force. Such a
suspension can be separated by simple filtration techniques. If the size of particles is less than
5 micron they undergo Brownian motion. In such suspension a stronger centrifugal force is
applied to separate the particles.

Centrifugal force is a pseudo force in a circular motion which acts along the radius and is
directed away from the centre of the circle. The force does not exist when measurements are
made in an inertial frame of reference. It only comes into play when changing our reference
frame from a ground/inertial to a rotating reference frame.

The centrifugal force’s unit is newton.

Figure No 1 Concept of centrifugal force

 With reference to the figure no 1 Considering a body of mass m rotating in a circular path of
radius r at a velocity of v. The force acting on the body in a radial direction is given by:
F = mv2/r
Where F = centrifugal force, m = mass of body, v = velocity of the body, r = radius of circle
of rotation.
The gravitational force acting upon the same body G = mg
Where, G = gravitational force
g = acceleration due to gravity
The centrifugal effect is the ratio of the centrifugal force and gravitational forces so that
(1)
Since, v = 2 r n where n = speed of rotation (r.p.m.)

(2)

where, k = 22/g = constant

D = maximum diameter of the centrifuge

D can be measured from the center of the centrifuge to the free surface of the liquid or to the
tip of the centrifuge tube.
From the equation C = kDn2 it can be concluded that
Centrifugal effect diameter of centrifuge
Centrifugal effect (speed of rotation)

Centrifugation
Centrifugation is the process that uses centrifugal force for the separation of two liquids in a
mixture. In this process, the denser component of the mixture migrates away from the axis
and the lighter component migrates towards the axis.
A centrifuge operates by using the sedimentation principle- Here the substances are
separated based on their density under the influence of gravitational force. When spun
rapidly, lighter particles stay at the top and heavier particles go to the bottom during
centrifugation.

Figure No 2 Concept of centrifugation

Factors that influence centrifugation:

 Density of both samples and solution

 Temperature/viscosity
 Distance of particles displacement
 Rotation speed

Q. 2 What are applications of centrifugation?

Ans - Centrifugation is a process which involves the use of the centrifugal force for the
sedimentation of heterogeneous mixtures with a centrifuge, used in industry and in laboratory
settings. This process is used to separate two immiscible liquids. More-dense components of
the mixture migrate away from the axis of the centrifuge, while less-dense components of the
mixture migrate towards the axis. Pharmacists, chemists and biologists may increase the
effective gravitational force on a test tube so as to move rapidly and completely cause the
precipitate ("pellet") to gather on the bottom of the tube. The remaining solution is properly
called the "supernate" or "supernatant liquid". The supernatant liquid is then either quickly
decanted from the tube without disturbing the precipitate, or withdrawn
If the particles of suspensions are very small then high centrifugal effect will be required to
separate the particles. To separate such suspensions the size of the centrifuge is kept smaller
but it is rotated at very high speed (rpm). If a large amount of material is to be separated and
a low centrifugal effect is sufficient to separate the suspension then the diameter (D) of the
centrifuge is increased and speed (n) is kept low.

Centrifugation is helpful in

1. Production of bulk drugs

After crystallization the drugs are separated from the mother liquor by centrifugation. For
example traces of mother liquor is separated from aspirin crystals by centrifugation
method.
2. Production of biological products
Most of the biological products are either proteinaceous or macromolecules. During
manufacturing they remain in colloidal dispersion in water. By normal methods of
filtration it is difficult to separate the colloid particles. In those cases centrifugal methods
are used.. Bacterial enzymes are separate from bacterial culture medium by sedimenting
the bacterial cells by centrifugation. Dirt and water are separated from olive oil and fish-
liver oils.
3. Determination of molecular weight of colloids
Polymers, proteins and such macromolecules often form colloidal dispersions. The
molecular weights of those molecules can be determined by ultracentrifugation.

Q.3 Classify centrifuges

Ans - Centrifuges can be classified on the basis of :
1) Speed
2) Temperature
3) Uses

1) On the basis of speed: The "speed" of a centrifuge is measured in rotation per minute,
or rpm. Centrifuges are generally divided into 3 categories based on their maximum
attainable speed: Low-speed : to maximum of 5 x 103 rpm.
High-speed : to maximum of 2 x 104 rpm.
Ultracentrifuges : to maximum of 105 rpm.
2) On the basis of temperature :
Refrigerated centrifuges have a built-in refrigeration unit surrounding the rotor, with
a temperature sensor and thermostat permitting selection of a particular temperature
or a permissible temperature range that is maintained during centrifugation. Many
biological samples are temperature sensitive, and centrifugation in the cold (1-4oC) is
frequently required.
Centrifuges that are not refrigerated are normally used at whatever temperature the
room they are in happens to be. This is typically described in research reports as
"room temperature" or “ambient temperature”.
3) On the basis of use :
Centrifuges Laboratory equipment
1) Swing-out arm type
2) Angle type
3) Ultracentrifuge
Commercial equipment --Perforated bowl types-
i) Batch- Top and Bottom driven
ii) Semicontinuous
iii) Ultracentrifiges
Sedimentation centrifuges
i) Vertical
a. Simple bowl
b. Bowl with plates
ii) Horizontal continuous decanters
Batch 1. Top driven 2. Under driven Semi continuous Continuous or Super centrifuge

 Principles, construction, working, uses, merits and demerits of

Perforated basket centrifuge,
Non-perforated basket centrifuge,
semi continuous centrifuge & super centrifuge

Q. 4 Explain principles, construction, working, uses, merits and demerits of

Perforated basket centrifuge,
Ans -
Bottom driven
 Figure 3 Perforated basket centrifuge (Bottom driven)

Principle
In basket centrifuges, the solids & liquids are separated by centrifugal force using a filter
media (usually a cloth) mounted over supporting mesh, which are together supported inside
the rotating basket. The slurry to be filtered is fed through the feed nozzles to the basket and
due to centrifugal force the liquid is forced out through the filter media while solids are
retained within the filter media inside the basket. These solids then separated or discharged
by various discharging methods namely - manually, bag lifting basket, through scrapper,
operated manually pneumatically / hydraulically, for which different models are available
Construction
The main parts of the Centrifuge are Bearing and Shaft, Basket, Drive and Brake
BEARING AND SHAFT: The Heavy duty bearing is chosen to withstand vibratory load and
centrifugal force and the shaft is made up of suitable material and duly balanced
independently without the basket for higher accuracy.
BASKET: The basket is made up of adequate thickness so as to withstand the loads caused
by the centrifugal forces.
DRIVE: The drive consists of Motor mounted at the basket casing driven through V belt with
provision of the tensioning the belts.
BRAKE: The Centrifuge are fitted with external shoe brakes with brake liners.
Working
The most common basket centrifuges utilize a perforated rotating basket and a filter media.
The feed material flows into the rotating basket and through the filter material under
centripetal force. Solids are retained in the basket and are removed either manually by
removing the filter bag or automatically with a peeler mechanism (scrapping) These
centrifuges are very economical to separate particles larger than .01mm and can produce very
dry cake. Additional wash cycles can be employed on the cake to remove original liquid
media. The centrifuge could be bottom top driven Figure 1 and 2 respectively.

Figure 4 Perforated basket centrifuge (Top driven)

Uses- 1) Can be used for thickening of suspensions/ slurries and also dewatering of biological
and fine solid sludge.
1) Used in waste water management to dewater the sludge obtained after pre-treatment.
 2) Can be used in variety of applications within the manufacturing and chemical
industry.
3) Can skim off oily contaminants
Advantages –
1) Same machine can be used for thickening as well as dewatering
2) Flexible in meeting process requirements
3) Little operator attention is required
4) Has skimming ability to separate oily contaminations
5) Full automation is possible
6) Clean operation
Disadvantages
1) It is not continuous operation
2) Requires special structural support unlike solid bowl centrifuge
3) Capacity is less
4) Lots of noise created during operation
5) Bowl needs washing between the shifts

Q 5 Explain principles, construction, working, uses, merits and demerits of Non

perforated basket centrifuge or Imperforate Basket Centrifuge

This centrifuge follows sedimentation mechanism without the help of porous medium.
Centrifugation follows the order of different densities between the solid and liquid phases.
Non perforated basket centrifuge principle
This centrifuge doesn’t contain any porous medium. The bowl contains a non-perforated
side wall. During centrifugation, liquid phase reach at top of the bowl and then collected.
But the solid phase retains on the side of bowl.
At the last minute of operation liquid phase is removed by skimming tube.

Fig 5. Non perforated centrifuge

Construction
 It consists of basket, which made of steel or other suitable materials.
 The basket is in vertical shaped, which is driven by a motor by using
suitable power.
 A feed pipe is adjusted to the basket.
  A skimming tube is used to collect the liquid from the top of basket.
 Scraper or sharp materials is used to collect the solid cakes from the side of
perforated wall.
Working
 Particles or suspension is introduced to basket by help of feed pipe.
 Start the motor by using suitable power system.
 Rotate the basket in maximum speed for specific period of time.
 Make sure the liquid phase float on the top of basket, while the solid phase
retains on the wall of basket. Then stop the motor.
 Collect the liquid by using the skimming tube.
 And collect the solid phase by using a scrapper or blade by hand.
Uses
 Chemical industry: Separating solids and liquids.
 Pharmaceutical industry: Purifying active ingredients.
 Food and beverage industry: Clarifying beverages and separating solids.
 Wastewater treatment: Reducing sludge volume and material recovery.
 Mining industry: Dewatering and clarifying mineral mixtures.
 Biotechnology industry: Biomolecule separation and concentration.
Advantages of Non perforated basket centrifuge
 Increased solids recovery.
 Improved product quality.
 Reduced operating costs.
 Lower maintenance requirements.
 Versatility in various industries and solid types.
Disadvantages
Here are some disadvantages of non-perforated basket centrifuge:
 Lower processing capacity.
 Longer processing times.
 Higher energy consumption.
 Limited solids discharge options.
 Higher initial cost.

Q 6. Explain principles, construction, working, uses, merits and demerits of

Semi-continuous Tubular Centrifuge

Principle- Semi-continuous centrifuges are long and thin enabling rapid acceleration and
deceleration, minimising the down-time required for the removal of the sedimented solids.
Here the radius and effective liquid thickness are both small allowing a high angular velocity
and hence high centrifugal force; small models can be used at acceleration factors up to
50,000 g, accumulating 0.1 Kg of wet deposit whereas large models, designed to accumulate
up to 5 Kg of deposit, are restricted to 16,000 g. The capacities of these centrifuges are only
moderate.

Construction and working -The Semi-continuous Tubular Centrifuge consists of machine

body, driving part / motor , tubular bowl, liquid collecting pan etc.

The material is sprayed onto the bowl from the inlet on the bottom and strong centrifuge
force is formed when the bowl rotates with the material and that makes the material flow
upper along the wall of the bowl and then separated because of the different gravity by the
centrifuge force.
Heavy liquid forms outer ring and light liquid forms inner ring. They flow to the top of the
bowl and discharge from their respective outlet. And the little solid is settled on the wall of
the bowl and discharged manually after the machine stops.

Fig 6. Semi-continuous Tubular Centrifuge

Uses-
It is mainly used for the liquid liquid separation or liquid liquid solid separation. For
examples:
1) Separation of blood (plasma and haemoglobin separation)
2) Vegetable and animal oil separation (vegetable oil degum, de-soap, fish oil polishing
3) Extraction and separation of coconut oil from coconut milk, essential orange oil, essential
rose oil etc
4) Waste water handling, waste oil treatment
5) Milk separation, coconut oil separation
Advantages
Strong in separation, simple in structure, convenient in operation and maintenance, low in
consuming power, small in ground, high in speed ,strong in separation factor, small in noise,
adaptive in material with high or low temperature.
Disadvantages-
Can not be used for heavy slurries, any solids settled must be removed by stopping the
machine
 Q 7 .Explain principles, construction, working, uses, merits and demerits of
Ultracentrifuge / super centrifuge
Principle- The ultracentrifuge is a centrifuge optimized for spinning a rotor at very high
speeds, capable of generating acceleration as high as 1 000 000 g (approx. 9 800 km/s²).There
are two kinds of ultracentrifuges, the preparative and the analytical ultracentrifuge. Both
classes of instruments find important uses in molecular biology, biochemistry,
and polymer science. Ultracentrifuges are laboratory centrifuges with rotors that spin at very
high speeds, usually ranging from 60,000 RPM and 200,000 x g to 150,000 RPM and
1,000,000 x g.
Ultracentrifuges are segmented into two main classes,
Preparative-- Preparative ultracentrifuges isolate or pellet biological particles, viruses,
organelles, membranes and biomolecules such as DNA, RNA and lipoproteins.
Analytical. --Analytical ultracentrifuges use detection systems to monitor spinning samples
in real time to determine sedimentation velocity and equilibrium used to determine shape and
mass of macromolecules.
Construction, working of Analytical ultracentrifuge
In an analytical ultracentrifuge, a sample being spun can be monitored in real time through an
optical detection system, using ultraviolet light absorption and/or interference optical
refractive index sensitive system. This allows the operator to observe the evolution of the
sample concentration versus the axis of rotation profile as a result of the applied centrifugal
field. With modern instrumentation, these observations are electronically digitized and stored
for further mathematical analysis. Two kinds of experiments are commonly performed on
these instruments: sedimentation velocity experiments and sedimentation equilibrium
experiments.
 Fig 7. Ultracentrifuge

Uses - Sedimentation velocity experiments aim to interpret the entire time-course of

sedimentation, and report on the shape and molar mass of the dissolved macromolecules, as
well as their size-distribution.

Sedimentation equilibrium experiments are concerned only with the final steady-state of the
experiment. This experiment is insensitive to the shape of the macromolecule, and directly
reports on the molar mass of the macromolecules

Analytical ultracentrifugation has recently seen a rise in use because of increased ease of
analysis with modern computers and the development of software.

SUPERCENTRIFUGE
Super centrifuge is a continuous centrifuge used for separating two immiscible liquid phases.
Principle:
 It is a sedimentation centrifuge.
 The separation is based on the difference in the densities between two immiscible liquids.
Centrifugation is done in the bowl of small centrifuge.
 During centrifugation, the heavier liquid is thrown against the wall, while the lighter liquid
remains as an inner layer.
 The two layers are simultaneously separated using modified weirs.

Fig 7. Super Centrifuge

Construction:
 It consists of a long hollow cylindrical bowl of small diameter.
 It is suspended from a flexible spindle at the top and guided at the bottom by loose-fit
bushing.  Two liquid outlets are provided at different heights at the top of the bowl, for
simultaneous recovery of the separated liquids using modified weirs.

Working:
 The centrifuge is allowed to rotate on its longitudinal axis at a high frequency usually about
2000 revolutions per minute with the help of drive-assembly.
 The feed is introduced from the bottom of the centrifuge using a pressure system.
 During centrifugation, two liquid phases separate based on the difference in their densities.
 The heavier liquid is thrown against the wall, while the lighter liquid forms an inner layer.
Both liquids rise to the top of the vertical bowl.
 The liquid-liquid interface (the so-called neutral zone) is maintained by an hydraulic
balance.  These two layers are simultaneously separately removed from different heights
through modified weirs.
 Thus, the super centrifuge can work for continuous separation of immiscible liquid phases.
Uses
 Preparative rotors are used in biology for pelleting of fine particulate fractions, such
as cellular organelles (mitochondria, ribosomes) and viruses.
 They can also be used for gradient separations, in which the tubes are filled from top
to bottom with an increasing concentration of a dense substance in solution. Sucrose
gradients are typically used for separation of cellular organelles.
 Gradients of caesium salts are used for separation of nucleic acids. After the sample
has spun at high speed for sufficient time to produce the separation, the rotor is
allowed to come to a smooth stop and the gradient is gently pumped out of each tube
to isolate the separated components.