CENTRIFUGATION

COURSE LEARNING OBJECTIVES

At the end of this topic, student will acquire the

ability:

1) To describe the basic types of centrifuges.

2) To determine the settling velocity of particles
both under gravity and under centrifugal forces.
3) To perform engineering analyses and scaling
calculations on tubular bowl and disk-stack
centrifuges.
 INTRODUCTION
 A centrifuge is used to separate particles or even
macromolecules, e.g.: cells, sub-cellular components,
proteins, nucleic acids.
 Basis of separation is according to the size, shape and
density of the component to be centrifuged.
 Centrifugation utilizes the density difference between
the solids and the surrounding fluid.
 When a suspension is allowed to stand, the denser
solids slowly settle under the influence of gravity, the
process is called sedimentation.
 In centrifugation of biological solids such as
cells, the particles are very small, the
viscosity of the medium can be relatively
high and the particle density is very similar
to the suspending fluid.
 The methodology of centrifugation:
i. Utilizes density difference between the
particles/macromolecules and the
medium in which these are dispersed.
ii.Dispersed systems are subjected to
artificially induced gravitational fields,
which amplified the settling rates.
 Densities of biological material
Material Density (g/cm3)
Microbial cells 1.05 - 1.15
Mammalian cells 1.04 - 1.10
Organelles 1.10 - 1.60
*Proteins 1.30
*DNA 1.70
*RNA 2.00
* Macromolecules are significantly smaller than the cells,
they would settle at extremely low velocities under
gravity and hence separation would not be feasible.
Therefore, they are separated by ultracentrifugation.
 Principle of centrifugation

Centrifuges are classified into two categories:

1) Laboratory centrifuges
2) Preparative centrifuges
Laboratory Centrifuges

Figure 1: Laboratory centrifugation

 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 to 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.
 The G value is also referred to as the RCF
(relative centrifugal force) value and it depends on
the rotation speed as well as the manner in which
the centrifuge tubes are held by the rotor:

r r ( 2n )
2 2
G  ------ (2.1)
g g

where:
r = distance from the axis of rotation (m)
 = angular velocity (radians/s)
g = acceleration due to gravity (ms-2)
n = rotation speed (s-1)
 G value in a centrifuge will depend on the
location:
 Highest: bottom of tube
 Lowest: top of tube
This implies that a particle will experience
increasing G values while moving towards the
bottom of the centrifuge tube.
Average value is frequently used for process
calculations.
Typical G values: 1,000 – 20,000.
 Rotors

Figure 2: Fixed angled and swing-out rotors

 Fixed angled rotors have a lower k-factor: smaller
difference between rmax and rmin
 Hence, the 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 with high sedimentation coefficient
such as cells and coarse particles.
 Precipitated macromolecules and finer particles
are centrifuged using fixed angled rotors.
 Preparative Centrifugation
Handle larger liquid volumes (i.e.: 1 to several thousand
litres).
Come in a range of designs.
Common feature: tubular 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 bowl centrifuge (Figure 3).
Alternative centrifuge: disk-stack bowl centrifuge (Figure
4) and ultracentrifuge.
Typical rotating speed: 500 - 2000 rpm.
Figure 3: Tubular-bowl centrifuge
The product stream enters at the bottom of the
centrifuge.
High centrifugal forces act to separate out the solids
and the bulk of the solids will adhere to the walls of
the bowl.
The liquid phase exists at the top of the centrifuge.
This type of centrifuge lacks a provision for solid
rejections, so the unit must be stopped in order to
remove the solids from the walls.
Suitable for particles with low sedimentation
coefficient, e.g. protein precipitates
Figure 4: Disk-stack bowl centrifuge
 A solid bowl holds a stack of conical plates that
act to decrease the settling distance while
simultaneously increasing the settling area.
The stacking of the plates assists in the rapid
sedimentation of the solids, and the stacking
angle allows the solid particles to slide smoothly.
The feed is introduced at the top of the bowl
and then travels through discs.
As they traverse through the unit, solids
become deposited on the underside of the discs.
Any remaining liquid is discharged through an
annular slit at the top.
Large particles have higher settling velocities than small
particles. Cellular debris ends up at the outer edge of the
bowl. Soluble intracellular material passes through with the
clarified liquid. Discs give a higher sigma factor.
1) The discs split the
stream into a large Benefit of Discs
number of very thin
layers thereby
improving
separation

2) Solids flow
downwards on
bottom face of disc

3) Liquid flows
upwards on top face
of disc

4) Sigma factor 
no. of discs
 Comparison of preparative centrifuges
System Advantages Disadvantages
Tubular 1) High centrifugal 1) Limited solids
bowl force capacity
2) Good dewatering 2) Foaming unless
3) Easy to clean special skimming or
4) Simple dismantling centripetal pump
of bowl used
3) Recovery of solids
difficult

Disk 1) Solids discharge 1) Poor dewatering

centrifuge possible 2) Difficult to clean
2) Liquid discharge
under pressure
eliminates foaming
3) Bowl cooling possible
 Ultracentrifugation
 Special type of centrifuge, the rotor rotates at a much
higher speed than the standard centrifuge.
 Typical rotation speeds: 30000 - 50000 rpm
 Ultracentrifuges:
1) Analytical ultracentrifuge (AUC) is mainly used for
studying properties of macromolecules.
2) Preparative ultracentrifuges are used to separate
macromolecules such as proteins and nucleic acids.
 The high speeds used in such devices generate
considerable amounts of heat.
 Therefore cooling arrangements are required in
ultracentrifuges.
 Settling of Solids
 The major forces acting on a solid
particle settling in a liquid by
gravitational forces are: FB + FD
i) gravitational force, FG
ii) drag force, FD
iii) buoyant force, FB
 When the particles reach a
terminal settling velocity, forces
acting on a particle balance each
other, resulting in a zero net force, FG
i.e.:

FG = FD + FB ------ (2.2)
where:


FG  D 3p  p g ------ (2.3)
6

FB  D 3p  f g ------ (2.4)
6
CD ------ (2.5)
FD   f 2 A
2

FD = drag force exerted by the fluid on solid particles

CD = drag coefficient ; Dp = diameter of particle
f = fluid density; p = particle density
 = terminal velocity of a particle
A = cross-sectional area of the particles perpendicular to the
direction of fluid flow; for a sphere, A = (/4)Dp2
 In general, the centrifugation of biological solutes
occurs within the Stokes regime, i.e. Re < 0.4.

 Drag coefficient for spherical particles is:

24 ------ (2.6)
CD 
Re

Re is the Reynold’s  f Dp ------ (2.7)

Re 
number: 

 Therefore, by substituting Eq. (2.6) and (2.7) into (2.5),

the drag force is given by Stoke’s Law:

FD  3D p ------ (2.8)

 Both the gravitational and buoyancy forces can be
written as follows:


FG  FB  D3p ( p   f ) g ------ (2.9)
6
 Substitution of Eq. (2.8) and (2.9) into Eq. (2.2) results in:


3Dp 
6

D p   f g
3
p 
p   f 2
or: g  Dp g ------ (2.10)
18
g is the terminal velocity during gravity settling of a
small spherical particle in dilute suspension.
 In a centrifuge, the corresponding terminal velocity is:

p   f 2 2
c  Dp  r ------ (2.11)
18
where:
c = particle velocity in the centrifuge
 = angular velocity of the bowl (rad/s)
r = radius of the centrifuge drum

 The ratio of velocity in the centrifuge to velocity under

gravity is called centrifuge effect or G value (Eq. 2.1):

r 2
G
g
 Performance of centrifuges of different size can be
compared using a parameter called the sigma factor, .
  represents the cross-sectional area of a gravity settler
with the sedimentation characteristics as the
centrifuge.
 For continuous centrifuges,  is related to the feed
rate of material as follows:

Q
 ------ (2.12)
g
where:
Q = volumetric feed rate
g = terminal velocity of the particles in a
gravitational field
 If two centrifuges perform with equal effectiveness:

Q1 Q2
 ------ (2.13)
1 2

where subscripts 1 and 2 denote the two centrifuges.

 Eq. (2.13) can be used to scale-up centrifuge equipment.

 Equations for evaluating  depend on the centrifuge
design.
 For a tubular-bowl centrifuge:

 Assume that particle is

located at distance z from the
bottom of the centrifuge.
 It is also located at position
r from the axis of rotation.
 This position is between the
liquid surface, R1 and the
bowl radius, R0 .

Figure 5: Idealization of the

tubular-bowl centrifuge
 The particle is moving in both the z and r directions.
 Its movement in the z direction comes from convection
of the feed pumped in the bottom of the centrifuge:

dz Q
 ------ (2.14)
dt  ( R0  R1 )
2 2

where Q is the feed flow rate. Effect of gravity at z

direction is negligible. Also assume that the centrifugal
force is so high that the liquid interface R1 is constant,
independent of z.
 The particle movement in the r direction is
related to its radial position, r :

dr  p   f 2 2
 Dp  r ------ (2.11)
dt 18
 In terms of the velocity of a particle settling under
the influence of gravity:

dr  r 2

 g   ------ (2.15)
dt  g 
where g is the velocity given by Eq.(2.10).
 Combining Eq. (2.11) and (2.15) to find the
trajectory of particle within this centrifuge gives:

dr dr dt  r 2   (R02  R12 )
  g   ------ (2.16)
dz dz dt  g  Q
 If g is large, the particle will quickly reach the wall.
 If Q is increased, the particle will be swept farther up
the tube.
 For the particles that are most difficult to capture, these
particles enter the tube at r = R1 and do not reach r = R0
until the end of the unit, i.e.: at z = l (i.e.: length of
bowl).
 Integration of Eq. (2.16) for these hard-to-catch
particles gives the maximum flow possible in the
centrifuge as a function of both particle properties
(g) and centrifuge characteristics (l, R0, R1 and ),
as below:

 l ( R 02  R 12 ) g  2 ------ (2.17)
Q 
g ln( R 0 R 1 )

 Because R0 and R1 in a tubular-bowl centrifuge are about

equal, Eq. (2.17) can be approximated as:

 2 lR 2 2  ------ (2.18)
Q  g    g
 g 
where R is the average radius roughly equal to R0 or R1.
  For a disk-stack bowl centrifuge:

Objective: to find the

position of a given particle
as a function of time.
 Consider a particle located
at position (x, y):
x = distance from the edge of
the outer discs along the
gap between the discs,
y = distance normal to the
lower disc
R0 = outer edge of discs
R1 = inner edge of discs

Figure 6: Idealization of disk centrifuge

 Liquid is fed into the centrifuge, it flows upward
through the gap between the discs, entering at R0
and leaving at R1 .

 The particle is moving both in the x and y directions. Its

velocity in the x direction is due to convection and to
sedimentation:

dx
 0 c sin ------ (2.19)
dt
where:
0 = convective liquid velocity
c = particle’s velocity under centrifugation
 = angle at which the discs are tilted from vertical
 The convective liquid velocity, 0 can be defined as
follows:

 Q 
0    f ( y) ------ (2.20)
 n ( 2rl ) 
where:
Q = total flow
n = no. of discs
r = distance from the axis of rotation
l = distance between the discs (measured normal
to the disc surfaces)
f(y) = some function giving the velocity variation
across the distance between discs.
 From a mass balance, the volume of 0 averaged
over y must equal to this convective velocity:

1 l  Q 
l 0
 0 dy   
 n ( 2rl ) 
------ (2.21)

 The resulting integration gives:

1 l

l 0
f ( y ) dy  1 ------ (2.22)
 Combining Eq. (2.19) and (2.20):

dx
  0   c sin 
dt
 0
------ (2.23)
 Q 
  f ( y)
 n(2rl ) 

This implies that the convective velocity is much greater

than that of sedimentation.
 Consider motion in the y direction. From Figure 6:

dy
  c cos  ------ (2.24)
dt
 From Eq. (2.10) and (2.11), the above equation can be
rewritten as:

dy   2r 
  g   cos  ------ (2.25)
dt  g 
 Combining Eq. (2.25) with Eq. (2.23):

dy dy dt

dx dx dt
------ (2.26)
 2nl  g  2
 2
  r cos 
 Qgf ( y ) 
 

 From Figure 6, it is noted that r = (R0 –x sin)2, hence:

dy  2nl   2


g
( R0  x sin  ) 2 cos  ------ (2.27)
dx  Qgf ( y ) 

which is the trajectory of a particle between the discs and

this centrifuge.
 Similar to tubular bowl centrifuge, consider the
particles that are most difficult particles to
capture.

 These particles enter at the outer edge of the discs,

where y = 0 and x = 0.

 They are captured at the inner edge of the discs,

at y = l and x = (R0 - R1)/sin .

 After capture, they and other particles are forced

along the disc surface to the outer edge, where they
are discharged.
 The integration of Eq. (2.27) for these hard-to-capture
particles and after rearrangement gives:

 2n 2 3 
Q  g   
R0  R1 cot 
3

 3g  ------ (2.28)
  g 

 In both Eq. (2.18) for tubular bowl and (2.28) for disk
centrifuge, the quantity g is only for the particle
properties, not for the centrifuge.
 The quantity in square brackets  has dimensions of
(length)2, is not the function of particle properties, but
reflects the geometry of the centrifuge.
 A higher  factor implies a higher centrifugal force,
leading to a more efficient separation process.
EXAMPLE 4:

A laboratory bottle centrifuge is used to collect yeast cells

after fermentation. The centrifuge consists of a number of
cylinders rotated perpendicular to the axis of rotation.
During centrifugation, the distance between the surface of
liquid and the axis of rotation is 3 cm, and the distance from
the bottom of the cylinder to that axis is 10 cm. The yeast
cells can be assumed to be spherical, with a diameter of 8.0
m and a density of 1.05 g/cm3. The fluid has physical
properties close to those of pure water. The centrifuge is to
be operated at 500 r/min. How long does it take to have a
complete separation?
SOLUTION:

From Eq. (2.11):

dr  p   f 2 2
 Dp  r
dt 18
We are interested in the yeast cell which takes longest to
settle, which is that starting near the liquid surface, i.e:

t=0 r = 3 cm

Integrating the initial equation gives:

r p   f 2 2
ln  Dp  t
3 cm 18
Inserting the values given:

g
0.05 3 2
10 cm cm 4 2  500 x 2π 
ln  (8 x 10 cm)   t
3 cm 18(0.01 g/cm sec)  60sec 
t  2500 sec

Therefore, this separation takes about 40 min to complete.

EXAMPLE 5:

A continuous disk-stack bowl centrifuge is

operated at 5000 rpm for separation of baker’s yeast.
At a feed rate of 60 liters min-1, 50% of the cells are
recovered. At constant centrifuge speed, solids
recovery is inversely proportional to flow rate.
a) What flow rate is required to achieve 90% cell
recovery if the centrifuge speed is maintained at 5000
rpm?
b) What operating speed is required to achieve 90%
recovery at the feed of 60 liters min-1?
SOLUTION:

a) If solids recovery is inversely proportional to feed rate,

the flow rate required is:

50%
(60 l min1 )  33.3 l min1
90%

b) Eq.(2.13): Q1 Q2

1 2
relates operating characteristics of centrifuges
achieving the same separation. From (a), 90% recovery is
achieved at Q1 = 33.3 l min-1 and 1 = 5000 rpm.
Q2 = 60 l min-1.
From Eq.(2.13),

Q1  1 33 . 3 l min  1
  1
 0 . 56
Q2  2 60 l min

Because the same centrifuge is used and all the geometric

parameters are the same, from Eq. (2.16):

dr dr dt  r 2   ( R02  R12 )
   g   ------ (2.16)
dz dz dt  g  Q

Hence, we can get this 1  2

correlation:   0.56
1
2  2
2
Therefore:

 2
(5000 rpm ) 2
 
2
2  1
 4 .46 x 10 7 rpm 2
0 .56 0 .56

Taking the square root:

2  6680 rpm
EXAMPLE 6:

Chlorella cells are being cultivated in an open pond. We

plan to harvest this biomass by passing the dilute stream of
cells through an available disc bowl centrifuge. The settling
velocity, g for these cells has been measured as 1.07 x 10-4
cms-1. The centrifuge has 80 discs with an angle of 40, an
outer radius of 15.7 cm and an inner radius of 6 cm. We
plan to operate the centrifuge at 6000 rpm. Estimate the
volumetric capacity, Q for this centrifuge.
SOLUTION:

Given:

g = 1.07 x 10-4 cm/sec

n = 80 discs
R0 = 15.7 cm
R1 = 6 cm
 = 40

Eq. (2.28) can be used to estimate Q:

 2n2 3 3 
Q  g  R0  R1 cot 
 3g 
 Substituting the values given into Eq. (2.28):

2
2n 2
2 (80 )  2 ( rad/rev )( 6000 rev / min) 

3g 3(980 cm / sec 2 )  60(sec/min ) 

 67497cm -1

R 3
0 R 1
3
cot   (15 .7 cm ) 3 3

 ( 6 cm ) cot( 40 )


 4354 . 5 cm 3
Q  1.07 x 10
cm
4

sec
67497 cm 1 4354 .5cm 3 
3
cm
 3.14 x 10 4

sec
liters
 31
sec
Therefore, this centrifuge is adequate only for a small pond.
EXAMPLE 7:

A pilot-scale disc-stack centrifuge is tested for

recovery of bacteria. The centrifuge contains 25
discs with inner and outer diameters of 2 cm and
10 cm, respectively. The half-cone angle is 35.
When operated at a speed of 3000 rpm with a feed
rate of 3.5 liter/min, 70% of the cells are recovered.
If a bigger centrifuge is to be used for industrial
treatment of 80 liters/min, what operating speed is
required to achieve the same sedimentation
performance if the larger centrifuge contains 55 discs
with outer diameter 15 cm, inner diameter 4.7 cm
and half-cone 45?
SOLUTION:

For small-scale centrifuge:

D1 = 0.02 m  R1 = 0.01 m
D0 = 0.10 m  R0 = 0.05 m
n1 = 25 discs
1 = 3000 rpm
Q1 = 3.5 L/min
1 = 35

Large-scale centrifuge:
Q2 = 80 L/min
n2 = 55 discs
D0,2 = 0.15 m  R0,2 = 0.075 m
D1,2 = 0.047 m  R1,2 = 0.0235 m
2 = 45
2 = ????
 Since the dimension of particles does
Q  g  not change in both cases, therefore g
remains constant.

Q1 Q 2
Q  
1 2

2n 2 3
1 
3g
R0  R13 cot 
2
2 (25) 
 0.05  0.01 m x
rad 3000 rev 1 min  1
  2
3 3 3
x x
 m rev min 60s  tan 35
3 9.81 2 
 s 
 106.6 m 2
 Q2
 2  x Σ1
Q1
80
 x 106.6 m 2
35
 2436.5 m 2 Next, find 2.

2n222 3
2 
3g
R0,2  R13,2 cot2

3g2
2 
2n2 R03,2  R13,2 cot2
1/ 2
 m 1 1 
 3 x 9.81 2 x 2436.5 m x
2

 

x tan 45 x 3
 s 2 (55) (0.075) 3
 (0 .0235) 3
m 
rad rev 60 s
 658 x x
s 2π rad 1 min
 6286.7 rpm
 SUMMARY
 Centrifugation is a very powerful method for
removing insoluble matters from process streams.
 The efficiency or throughput achievable with
centrifugation improves with:
a) higher density differences between the particle and
the medium
b) larger particle sizes
c) lower liquid viscosities
 Centrifugal separation is an attractive method for solid-
liquid and liquid-liquid separations as continuous
processing is feasible, retention times can be short and
no filter aids are required.
EXERCISE:

1) A tubular bowl centrifuge is used to recover yeast cells

from a fermentation broth. At a flow rate of 10 liters/min
and 5000 rpm, 50% of the cells can be recovered. You are
now asked by your manager to increase the recovery to
90% using the same equipment.

a) What flow rate should you use?

b) If you double the rotation speed, how much does the
centrifugation velocity change?
2) A tubular bowl centrifuge is used to concentrate
a suspension of Escherichia coli prior to cell disruption. The
bowl of this unit has an inside
radius of 12.7 cm and a length of 73.0 cm. The speed of the
bowl is 16,000 rpm and the volumetric capacity is 200
liters/h. Under these conditions, this centrifuge works well.

a) Calculate the settling velocity, g for the cells.

b) After disruption, the diameter of debris is about one-
half of the original cell diameter and the viscosity is
increased four times. Estimate the volumetric capacity
of this same centrifuge operating under these new
conditions.
3) Yeast cells are to be separated from a fermentation broth.
Assume that the cells are spherical with diameter 5 m and
density 1.06 gcm-3. The viscosity of the culture broth is
1.36 x 10-3 Nsm-2. At the temperature of separation the
density of the suspending fluid is 0.997 gcm-3. 500 liters of
broth must be treated every hour.

a) Specify  factor for a suitably-sized disk-stack centrifuge.

b) The small size and low density of microbial cells are
disadvantages in centrifugation. If instead of yeast, quartz
particles of diameter 0.1 mm and specific gravity 2.0 are
separated from the culture liquid, by how much is 
factor reduced.