Mass Transfer

Transfer of mass from one location to another

Occurs in areas of concentration variation

Occurs till equilibrium is established

Occurs in many processes such as evaporation, adsorption,
drying, precipitation, filtration and distillation.




In bioprocess
Concentration of compounds are not uniform

Mass transfer principle is made use of to achieve this

E.g., Oxygen transfer
Solvent extraction

Aeration
Oxygen is normally supplied to microbial cultures in the form of
air, i.e cheapest source

Sterile air / oxygen which must be dispersed throughout
the fermenter

Air introduced into the fermentor is filter sterilized and
introduced via sparger which is located below the agitator

Sparger structure affects oxygen transfer in the medium as it
influences the size of the gas bubble produced

Smaller the bubble, larger the surface area to volume ratio
which provides greater oxygen transfer.

Spargers with small pore size are effective in producing small
bubbles but are prone to clogging and require high energy








Oxygen transfer
Oxygen transfer is complex and it involves a phase change
from its gaseous phase to the liquid phase which is influenced
by following factors

1. Temperature, pressure and surface area of oxygen bubbles
2. Chemical composition of the medium
3. Volume of gas introduced per unit reactor volume per
unit time
4. Type of sparger system used to introduce air into the
fermenter
5. Speed of agitation

In aerobic fermentation- oxygen should be maintained
at optimal concentration for maximal yeild



Oxygen mass balance

-The rate at which O
2
can be delivered to the biological
sysytem (OTR-Oxygen transfer rate) and the rate at which
it is utilised by microorganism (COD- Critical oxygen
demand)
Anaerobic conditions develop when the rate of Oxygen
utilization is > than OTR ( This limit growth and production)

OTR can be increased by elevating the pressure,
enriching the inlet air with O
2
and increasing agitation and
aeration
Solubility of oxygen depends on temperature
and pressure

Temperature
in C


p(O2)=100 kPa
Solubility in mg/l


p(O2)=20.9 kPa
Solubility in mg/l

Determination of OTR
OTR = Oxygen gradient

Resistance to oxygen transfer

= Oxygen gradient ( C*-C
L
)
eqn 1
K
L
a
C* =is the saturated dissolved oxygen conc.(mmol/dm
3
)
C
L
=is the concentration of dissolved O
2
at time (t) (mmol/dm
3
)
K
L
=is the mass transfer coefficient at the gas to liquid phase
(phase boundary)(cm/h)
a =




is the gas/liquid interface area per liquid volume (cm
2
cm
-3
)
Stages of resistance of oxygen transfer from gaseous phase
to an individual cell
1. Resistance within the gas film to the phase boundary

2. Penetration of the phase boundary between gas bubble and
liquid

3. Transfer from the phase boundary to the liquid

4. Movement within the nutrient solution

5. Transfer to the surface of the cell

6. Entry into the cell

7. Transport to the site of reaction within the cell
Gas bubble
Gas
film
Fluid
film
Fluid
Fluid
film
Rate of O
2
transfer from air bubble to the liquid phase is described as
dC
L
= k
L
a(C
*
- C
L
) eqn. 2
dt
Integration of eqn 1 gives
C*-C
L
= e
-KLat
eqn 3


Interms of natural log, for k
L
a determination



ln (C
*
- C
L
) = -K
L
a (t) eqn 4
K
L
a for the specific conditions is determined by plotting a semilog
graph of ln(C
*
- C
L
) against time where slope is mass transfer
coefficient (K
L
a)

Therefore K
L
a is a measure of the aeration capacity of a fermentor and
must be maintained above a minimum critical level to satisfy oxygen
requirements





K
L
a
K
L
= Is the mass transfer coefficient
a = Is the gas/liquid interface area per liquid volume (cm
2
cm
-3
)

These are difficult to measure individually and are generally
linked to give as

K
L
a = volumetric mass transfer coefficient per hr

How to improve the oxygen transport?
Increase of the O
2
-solubility
Pressure increase from 100 to 200 kPa
Increase the O
2
-content in the air
enrichment of the aeration with O
2

Use of pure O
2

Change in the phase boundary (gas/liquid)
size and distribution of the gas bubbles
contact time between the gaseous phase and the liquid
phase
Viscosity of the nutrient solution
viscosity reduction increases the relative velocity of the
gas bubbles thinner liquid film higher k
L
a-value



Scale-up
What is Scale-Up?
Increasing the scale of a fermentation. (i.e.
From lab scale to pilot scale and from pilot
scale to industrial scale
3 Stages
Bench Scale ( 2 20 L)
Pilot Scale (100 500 L)
Plant Scale (500 20,000 L)


What is a scale-up problem?
A scale-up problem is something that we do not see in
the small-scale experiment(lab scale) and are
surprised and disappointed to find in the large scale
process..

Problems associated with scale up are due to the
different ways in which process parameters are
affected by increase in size of the unit
1) Inoculum development
when a scale is increased -extra stages have to be
incorporated in the inoculum development programme

2)Sterilization
It is a scale dependent factor
When there is increase in scale of a fermentation process,
the sterilization regime should be adjusted according to the
scale
This may result in change in quality of the medium after
sterilization

Major factors involved in
Scale-Up
3. Environmental parameters
Increase in scale may results in a change in the environment
for the microorganism.
Environmental parameters may changed due to increase in scale:
i) Nutrient availability
ii) pH
iii) Temperature
iv) Shear conditions
v) Dissolved oxygen concentration
vi) Dissolved CO2 concentration
vii) Foam production




Agitation
Aeration
agi tati on
aerati on
shear
cost
foam
mi xi ng
oxygen
CO
2
Scale up window based on AGITATION /
AERATION
This illustrates the "scale-up" window defining the
operating boundaries for aeration and agitation in the scale-
up of a typical fermentation.

Agitation and aeration rate must fall between a minimum
and maximum value
Problems that may arise, when these values are not falling
within the limits.

ACTION RESULT
Minimise aeration below the limit Decrease in CO2 and
O2 levels
Maximise aeration above the limit Increased Foam formation takes
place
Minimise agitation below the limit Bulk mixing poor

Maximise agitation above the limit Shear and cost increased
Solution for scale up problem
Identify the environmental parameter affected by aeration and
agittaion. eg: Oxygen concentration, Shear, bulk mixing

Identify the process variable or variables which affects the
identified environmental parameter

Calculate the value of the process variable to be used on large
scale which results in the same environmental conditions on both
scales
Process variables which affect the mixing and mass transfer

Process Variable Mass transfer or Mixing
property affected
Power consumption per
Unit volume
Oxygen transfer rate
Volumetric air flow rate Oxygen transfer rate
Impeller tip speed Shear rate
Pumping rate Mixing time
Reynolds number Heat transfer
Scale-Up Parameters
First scale-up criterion is the preservation of
Geometrical similarity
when building a new bioreactor vessel, the geometry is
usually scaled linearly.
height to diameter ratio (H/D) or aspect ratio is kept
constant to ensure the tanks will operate similarly.


Vessel 1
Vessel 2
The aspect ratio of the vessels is 1.5.
The height and diameter between the two
vessels is scaled linearly.

By multiplying the dimensions of Vessel 1 by
6.4, the dimensions of Vessel 2 are determined.

Note: the volume of the vessels does not scale
linearly (volume of vessel 1 is 147 ft
3
and vessel
2 is 38,603 ft
3
, 262 times larger.)
To scale-up a manufacturing process from one
bioreactor to another, the process parameters are
scaled based on the following :

Agitation-based scaling parameters
Gassing-based scaling parameters

NOTE: Cannot keep all parameters constant during
scale up because they scale by different values




Scale-Up Parameters
Common Scale-Up Parameters
Agitation-based scaling parameters:
Mixing Time
Power Input per Volume (P/V)
Tip Speed
Notes: These three parameters are all dependent on agitation
rate, so all three cannot be held constant when scaling-up. For
example:
Keeping mixing time constant might cause a high
P/V that the cells cannot handle.
Scaling based on constant tip speed might cause a
low agitation rate that will not deliver oxygen
adequately.
- Thus all three scaling parameters must be
evaluated and the final scale-up agitation rate must
produce acceptable values for all three parameters.

Gassing-based scaling parameters
Vessel Volumes per Minute, VVM
Superficial Gas Velocity, V
s

Note: The two gas flow rate scaling parameters are both
dependent on the dimensions of the vessel. Scaling based on
one will greatly affect the other.
Scale-Up Parameters
Agitation Parameters
Parameter Definition Scale-Up Factor Why is this
Important?



Mixing Time


Amount of time it takes the
bioreactor to create a
homogeneous environment
N
2
=N
1
(D
1
/D
2
)
1/4
N
2
agitation speed in scale-up
N
1
agitation speed in scale-
down
D
1
impeller diameter of scale -
down
D
2
impeller diameter of scale-
up




Want to ensure that the
materials are well-mixed in a
timely manner

Power Input per
Volume (P/V)

Amount of power
transferred to a volume of
cell culture through the
agitator shaft and impellers
P/V N
3
/D
2
P- power supplied
V- Volume of Bioreactor
N- Agitation Speed
D- Impeller Diameter
Mammalian cells cannot
handle a lot of power
introduced into the culture
media as it can cause small
eddies that will shear the
fragile cell membranes


Tip Speed


Related to the shear rate
produced from the impellers
moving through the cell
culture media
N
2
=N
1
(D
1
/D
2
)

N
2
agitation speed in scale-up
N
1
agitation speed in scale-
down
D
1
impeller diameter of scale -
down
D
2
impeller diameter of scale-
up
High shear rates can cause
the cell membrane to tear
and the cells to die.
If scale-up based on
constant tip speed is
attempted, P/V and mixing
time will decrease
Gassing Parameters
Parameter Definition Scale-Up Factor Why is this
Important?

Vessel Volumes
per Minute
(VVM)
means the volume of
gas flow per bioreactor
volume per minute.


Volume of Gas Flow/time
necessary to ensure that
enough oxygen will be
supplied to the cells

Superficial Gas
Velocity (V
s
)


volume of gas per
cross-sectional area of
the vessel.
V
s
= Q
gas
/A
v

V
s
- superficial gas velocity
Q
gas
- gas volumetric flow rate
A
v
- inside cross-sectional area
of vessel
increasing V
s
causes

An increase in foam
generation
A decrease in P/V
An increase in oxygen
transfer

Scale-Up Review
Overall, scaling up process parameters is tricky

Each scale-up parameter is dependent on another.
Scaling-up based on constant P/V will affect the mixing
time and the tip speed in the bioreactor. As well, for gas
flow rates, scaling-up on constant V
s
will affect the VVM.

Cannot keep all constant during scale-up

Not one scale-up process is correct

Technicians determine which parameter is critical to the
process and try to find a happy medium between each of
the remaining parameter