Centrifugation

Centrifugation

When a solid-liquid suspension is rotated in a cylindrical

container (bowl) the suspension is subject to a centrifugal
force in the radial direction.
Centrifugation is a process by which solid particles separated
from a liquid using centrifugal force as a driving force.
The centrifugal force can be many times greater than the
force of gravity , allowing even very small particles or
particle slightly denser than the fluid to settle.
The rate of centrifugation is specified by the angular velocity
measured in revolution per minute (rpm), or acceleration
expressed as g.
The conversion factor between rpm and g depend on the
radius of the sample in the centrifugal rotor.
The particles setting velocity in centrifugation is a function
of their size and shape, centrifugal acceleration ,the volume
fraction of solid present, the density difference between the
particle and liquid, and viscosity.

Induced
gravitational
field

Supernatant

Precipitate
Suspension

In
process

Separation
complete

Centrifuges are classified into two categories:

Laboratory centrifuges
Preparative centrifuges

Laboratory centrifuges
Centrifuge
tube

Suspension

Supernatant

Precipitate

Rotor

Used for small-scale separation and particle free sample preparations

Typical liquid volumes handled is about 1 5000 ml
The material to be centrifuged is distributed in centrifuge tubes
Tubes are attached rotor in a symmetric manner
Two types of rotors: fixed rotors and swing out rotors
Induced gravitational field move particles towards the bottom of the tubes
Typical rotation speeds:1,000 15,000 rpm
Induced gravitational field is measured in terms of the G value
G value depends on the rotation speed as well as the manner in which the
centrifuge tubes are held by the rotor

Laboratory centrifugation
r 2 2rn 2
G

g
g
G value will depend on the location
Highest: bottom of tube
Lowest: top of tube
Particle experience variable G values during their motion
Average value is frequently used
Typical G values: 1,000 20,000

Rotors
Fixed angled rotors have smaller difference between rmax and rmin
Time required for precipitation is less with the fixed angled rotor
Distance travelled by particles is less with the angled rotor
Fixed angled rotors are heavier and require much higher energy to operate
Swing out rotors are preferred for centrifuging substances such as cells and
coarse particles
Precipitated macromolecules and finer particles are centrifuged using fixed
angled rotors
Fixed angled rotor

Rmax - Rmin

Swing-out rotor

Rmax - Rmin

Preparative centrifugation
Handle larger liquid volumes (i.e. 1 to several thousand litres)
Range of designs
Common feature: rotating chamber into which the suspension is fed from one
end while the supernatant and precipitate is collected from the other end in a
continuous or semi-continuous manner
Most common type: tubular centrifuge
Typical rotating speed: 500 - 2000 rpm
Supernatant
collection
Precipitate collection

Developing
precipitate
layer

Rotating tubular
bowl
Annular space

Supernatant

Feed

CENTRIFUGE

Ultracentrifugation:

Ultracentrifugation

DENSITY GRADIENT CENTRIFUGATION

Important technique for purifying proteins and

particularly nucleic acids.

Density gradient
centrifugation

DNA separation