Measurement of kLa
Static method (Oxygen-Balance Method)
Sulfite Oxidation Method
Dynamic Method
Static method (Oxygen-Balance Method):
This technique is based on the equation for gas-liquid
mass transfer .
N A k L a (C * C L )
In the experiment, the oxygen content of gas streams
flowing to and from the fermenter are measured.
From a mass balance at steady state, the difference in
oxygen flow between inlet and outlet must be equal to
the rate of oxygen transfer from gas to liquid:
NA is the total quantity of gas absorbed by the system
per unit time and per unit volume
where VL is the volume of liquid in the fermenter,
Fg is the volumetric gas flow rate,
CAG is the gas-phase concentration of oxygen, and
subscripts i and o refer to inlet and outlet gas
streams, respectively.
Since gas concentrations are generally measured as
partial pressures,
Advantage of this method is that the method can
be applied to fermenters during normal operation.
It depends on accurate measurement of gas
composition, flow rate, pressure and temperature
Sulfite Oxidation Method
It is referred as Chemical method(Cooper et al., 1944)
It is based on the oxidation of sodium sulfite to sodium sulfate
in the presence of catalyst (Cu++ or Co++).
This reaction has following characteristics to be qualified for
the measurement of the oxygen-transfer rate:
1. The rate of this reaction is independent of the concentration
of sodium sulfite within the range of 0.04 to 1 N.
2. The rate of reaction is much faster than the oxygen transfer
rate; therefore, the rate of oxidation is controlled by the rate of
mass transfer alone.
Procedure: Fill the fermenter with a 1 N sodium sulfite
solution containing at least 0.003 M Cu++ ion.
Turn on the air and start a timer when the first bubbles
of air emerge from the sparger.
Allow the oxidation to continue for 4 to 20 minutes,
after which, stop the air stream, agitator, and timer at the
same instant, and take a sample.
Mix each sample with an excess of freshly pipetted
standard iodine reagent.
Titrate with standard sodium thiosulfate solution
(Na2S2O3) to a starch indicator end point.
Once the oxygen uptake is measured, the kLa may be
calculated by using
rA= kL a (C*AL-CAL)
where CAL is zero and C*AL is the oxygen equilibrium
concentration.
Dynamic Method
It is based on an unsteady-state mass balance for
oxygen.
The main advantage of the dynamic method over
the steady-state technique is the low cost of the
equipment needed.
During the re-oxygenation step, the system is not
at steady state.
The rate of change in dissolved-oxygen
concentration during this period is equal to the
rate of oxygen transfer from gas to liquid, minus
the rate of oxygen uptake by the cells:
where qox is the rate of oxygen consumption.
We can determine an expression for qox by
considering the final steady dissolved-oxygen
concentration, CAL.
When CAt = CAL, - dCAL/dt= 0 because there is no
change in CAL with time.
Substituting this result gives
Problem#1
A 20L stirred fermenter containing a Bacillus thuringiensis
culture at 30°C is used for production of microbial
insecticide, kLa is determined using the dynamic method.
Air flow is shut off for a few minutes and the dissolved-
oxygen level drops; the air supply is then re-connected.
When steady state is established, the dissolved-oxygen
concentration is 78% air saturation. The following results
are obtained.
Problem# 2
Factors Affecting Cellular
Oxygen Demand
Bubble Size
Gas hold-up
Gas Velocity
Type of gas Sparger
Type of agitation
Power input to agitator
Temperature
Pressure
Antifoaming agents
Presence of cells
Surface-active solutes.
