CSB Chap 4
CSB Chap 4
CSB Chap 4
Downstream processing of
bio-product
(Chapter 4: Sedimentation)
Contact:
Mrs. Rozyanti Mohamad
06-551 2077 / 012-652 0389
[email protected]
Sedimentation
Sedimentation is the tendency for
particles in suspension to settle out of the fluid due to
gravitational or centrifugal acceleration
g = 9.81 ms-2
2R
R: distance of particle
from the center of
rotation
: angular velocity
(rad/s)
Equation of motion
Nearly all analyses of separation by sedimentation begin
with the equation of motion of a particle with:
oRadius,
oDensity,
oMass, m=(4/3)
i : inertial acceleration
b : buoyancy of particles
d : Stokes drag force
(Eq. 4.1)
Equation of motion
Velocity in a centrifugal field at steady state or
centrifugation equation, :
2a 2 ( 0 ) 2 R
=
......( Eq.4.2)
9
: dynamic viscosity
o
: medium density
Equation of motion
If the particle moving out, R is not constant (at t = 0, R =
R0).
The equation become:
R 2a 2 ( 0 ) 2t
ln =
......( Eq.4.4) Where t = time
9
R0
Creeping flow
o Creeping flow conditions are usually satisfied in
sedimentation, whose Reynolds number is very small
(Re < 1).
Re =
2a
......( Eq.4.5)
1
Layer the solution of
decreasing density
Starting from the
bottom until the vessel
is filled.
The resulting gradient is
like a staircase.
2
Centrifuge at extremely
high speed
Resulting in isothermal
stratification of densityforming solute.
3
Gradient mixing method
Consist of two
cylindrical container :
one containing
concentrated solution
and another containing
dilute solution
Equipped with stirring
apparatus and link each
other
produce outflow with
linear salt gradient.
Hindered settling
o When concentration of
sedimenting particles
increase, velocity
become decrease.
o In hindered settling,
particles are so close
and they have a major
effect on each other.
Sedimentation coefficient
o When a body force is applied, velocity through a viscous
medium is usually proportional to the accelerating field
o In the case of sedimentation, the resulting constant (a property
of both particle and medium) is sedimentation coefficient
defined as;
S 2
R
Where
= dR/dt
S = the Svedberg (1S = 10-13s)
Equivalent time
o First, we define G as the dimensionless acceleration, the ratio
of centrifugal to gravitational acceleration for a particular
centrifuge:
2R
G
g
Equivalent time
o A rough approximation of the difficulty of a given separation by
centrifugation is the product of the dimensionless acceleration
(G) and time required for separation (t), called as equivalent
time:
2R
Equivalent time Gt =
t ......(Eq .4.7)
g
Eukaryotic cells
Protein precipitates
Bacteria
Ribosome
= 0.3 x 106 s
= 9 x 106 s
= 18 x 106 s
= 1,100 x 106 s
Sigma analysis
o Commonly used in industry
o Use the operation constant () to characterize a centrifuge into
which feed flows at volumetric flowrate (Q):
Q = {g}{}
Where;
2a 2 ( 0 ) g
vg = sedimentation velocity at 1xg: g =
Sigma analysis
o The sedimentation velocity at 1xg can directly
determine using:
2a 2 ( 0 ) g
g =
9
R
g ln
R0
2t
Centrifuge:
Basis of separation
Basis of separation
Size
Shape
Density
Centrifuge:
Methods of separation
o Methodology:
o Utilizes density difference between the
particles/macromolecules and the medium in which these
are dispersed
o Dispersed systems are subjected to artificially induced
gravitational fields
Principle of centrifugation:
Stokes Law
The concept of using centrifugal force to efficiently separate
components of differing densities
The fundamental principle that applies to centrifugation is
Stokes Law
Formula for determining the rate of sedimentation
It states that a particle moving through viscous liquid attains a
constant velocity or sedimentation rate
The rate can be very slow for particles whose:
(1) density is close to that of the liquid
(2) for particles whose diameter is small
(3) where the viscosity is high
Principle of centrifugation:
Stokes Law
Stokes Law: =
d 2 2r (Pp Pi )
18
......(Eq .4.8)
Where:
= rate of sedimentation (cms-1)
D = particle diameter (cm)
= angular velocity about the axis(radianss-1)
r = distance of particle from the axis of rotation (cm)
Pp = density of particle (g cm-3)
Pi = density of liquid (g cm-3)
= viscosity of liquid (g cm-1sec-1)
Principle of centrifugation:
Centrifugal acceleration
Replacing gravitational acceleration with the acceleration
generated by a rotating centrifuge results in faster
sedimentation.
Centrifugal acceleration can be thousands of times greater
than that of gravity, so the centrifugal sedimentation rate is
thousands of times greater.
What is RPM..?
What is RCF..?
Principle of centrifugation:
Centrifugal acceleration
RPM
(Revolutions per
minute)
RCF
(Relative centrifugal
force) is measured in
force x gravity or gforce.
Principle of centrifugation:
Centrifugal acceleration
Relationship between RCF and RPM:
2
RPM
RCF = 11.2 r
1000
Where;
RCF
RPM
r
Exercise #1
The sedimentation coefficient for smaller ribosomes from E.coli
was determine at 70S. Estimate how long it would take to
completely clarify a suspension of these ribosomes in a high
speed centrifuge operating at 10,000 rpm with a tube containing
the ribosomes suspension in which the maximum distance of
travel particles radially outward is 1 cm and the initial distance
from the center of rotation to the particle nearest is 4 cm.
Exercise #1 (solution)
Based on the equation for sedimentation coefficient;
dR
dR 1
2
s=
s dt =
2
dt R
R
R
R0
Exercise #1 (solution)
To determine the maximum time required, we evaluate R at the
maximum travel of the cells from the center of rotation (5 cm)
R
1h
ln
(
)
ln
5
4
R
3600 s
t= 20=
= 8.1 h
2
s
rev 2 rad 1 min
13
x
x
10,000
70x10 s
min
rev
60 s
Exercise #2
If bacterial cell debris has Gt = 54 x 106 s, what is the
centrifuge speed needed for a full sedimentation in 2 h
of time using 10 cm diameter centrifuge?
Exercise #2 (solution)
From the equation for equivalent time:
1
2
m
2R
rad
Gtg 54 x 10 s x 9.81 s 2
Gt =
t=
= 1,213
=
g
s
Rt 0.05 m x 2(3600)s
1
2
rad 1 rev 60 s
= 1,213
x
x
s 2 rad min
= 11,590 rpm
Exercise #3 (solution)
1) Calculate the speed of rotation in RPM. Given:
(a) r = 30 mm, RCF = 6,000 x g
13,360 rpm
(b) r = 100 mm, RCF = 8,000 x g 8,452 rpm
2) Solve RCF value for:
(a) RPM = 40,000, r = 105.4 mm
(b) RPM = 80,000, r = 210.8 mm
188,877xg
1,511,014xg
Production centrifuge
The concept of centrifugal force (a.k.a centripetal force) can be
applied in rotating devices such as centrifuges, when they are
analyzed in a rotating coordinate system.
A centripetal force is a force that makes a body follow a curved
path.
Production centrifuges
Production centrifuges:
Tubular bowl centrifuge
Liquid enters the
bowl through an
opening in the center
of the lower bowl
head.
Particles are
centrifuged towards
the wall of the
cylinder
Production centrifuges:
Tubular bowl centrifuge
The equation of motion is used to determine
the flowrate (Q).
dR 2a 2 ( 0 ) 2 R
=
Radial direction:
9
dt
Production centrifuges:
Tubular bowl centrifuge
Integrating dR (between R0 and R1) and integrating dz
(between 0 and L) and solve for Q:
2a 2 ( 0 ) L( R02 R12 ) 2
Q=
R
9
ln 0
R1
L( R 2 R 2 ) 2
0
1
=
R
g ln 0
R1
Production centrifuges:
Disc bowl centrifuge
Feed suspension
enters on the axis
of rotation
Feed is forced to
the bottom of
rotating bowl
Pressure forces
the suspension
upward
Heavier fluid is
forced through
holes at the end
of each disc
Dense particles
are ejected
through a nozzle
Addition of flocculants :
Forming interparticle
molecular bridges
between particles
Reducing the
repulsive forces
between cells.
D =
kT
6 a
kT
v
D/L
Particle-free overflow
exits the upper end
Particle-rich suspension
leaves in the underflow
Centrifugal elutriation
Similar to inclined sedimentation and field-flow fractionate
(sedimentation in presence of fluid flow).
In a centrifugal elutriation (a.k.a counterstreaming
centrifugation), fluid is continuously pumped in the opposite
direction to that sedimentation.
Industrial application
Industrial centrifuges can be classified into two main types:
sedimentation and filtering centrifuges.
1. Sedimentation centrifuges
Use centrifugal force to separate solids from liquids, as well as
two liquids with different specific gravities (decanter, disk-stack,
solid-bowl basket and tubular bowl centrifuges)
2. Filtering centrifuges
Use centrifugal force to pass a liquid through a filtration media,
such as a screen or cloth while solids are captured by the filtering
media. Filtering centrifuges primarily deal with perforate basket,
pusher and peeler centrifuges
Industrial application
Wastewater processing
Deals with separation of municipal, farm, DAF (dissolved air
flotation), trap grease, drilling mud, and environmental
wastewater sludges.
Chemical processing
Which produces raw products such as acids, salts, oil refinery
by-products, polymers, oil-water-solids, and so on.
Pharmaceutical and Biotechnology industries
That manufacture drugs, vaccines, medicines, penicillin,
mycelia, E-coli bacteria, algae, enzymatic waste, etc.
Industrial application
Fuel and Biofuel industry
Including synthetic fuels, biodiesel, ethanol, cellulosic ethanol,
algae biomass dewatering; fuel and lube oil purification, etc.
Food Processing
Which deals with refining of vegetable oils, dairy (milk, cheese,
etc.); poultry, swine and beef rendering; yellow, white, and brown
grease separation; fruit and vegetable juice; beer, wine and liquor
clarification, etc.
Mining and mineral processing
Including coal, tar sands, copper, precious metals, calcium
carbonate, kaolin clay, and many more.
Industrial application
Sedimentation
principles
Equation of
motion
Gravitational
acceleration
Centrifugal
acceleration
Sensitivities
Reynolds
No.
Equilibrium
sedimentation
Production
centrifuge
Methods
Layer the
solution of
decreasing
density
Centrifuge
at high
speed
Gradient
mixing
method
Coefficients
Sedimentatio
n coefficient
Equivalent
time
Sigma
analysis
Tubular
centrifuge
Disk
centrifuge
Flocculation
and
sedimentation
Sedimentation
at low
acceleration
Diffusion,
Brownian
motion
Isothermal
settling
Convective
motion and
Peclet No.
Inclined
sedimentation
Field flow
fractionatio
n
Centrifugal
elutriation
Exercise #4
It is desired to achieve complete recovery of bacterial cells from
fermentation broth with a pilot plant scale tubular centrifuge. It
has been already determined that the cells a approximately
spherical with radius of 0.5 m and have density of 1.10 g/cm.
The speed of the centrifuge is 5000 rpm, the bowl diameter is 10
cm, the bowl length is 100 cm, and the outlet opening of the
bowl has a diameter of 4 cm. Estimate the maximum flowrate of
the fermentation broth that can be attained (viscosity of water is
0.01 g/cm.s).
Exercise #4 (solution)
The flow rate can be estimated by:
Q = {g}{}
where
6
3
g
m
10
cm
2 0.5x106 m x (1.10 1.00) 3 x 9.81 2 x
2
3
2 ( 0 )g
cm
s
m
g =
=
g
9
9 0.01
cm s
cm
= 5.45x106
s
2
Exercise #4 (solution)
For complete recovery of tubular bowl centrifuge;
L( R 2 R 2 ) 2
0
1
=
R
g ln 0
R1
rev 2 rad
(100 cm )x 52 2 2 cm 2 x 5000
x
min
rev
=
2
m
5 100 cm 60 s
9.81 2 x ln x
x
s
2
m
min
= 2.01 x 106 cm 2
Exercise #4 (solution)
Therefore,
Q = {g}{}
cm
L
60 s
6
2
= 5.45x106
x
2.01x10
cm
x
x
s
103 cm3 min
= 0.66
L
min