Experimental Techniques for kLa determination

http://www.youtube.com/watch?v=8441Bz07qQ0
Theory: Narasimha

dC AL
dt
(
= k L a C *AL − C AL )
integrating….

C AL 2 t
1
∫
C AL1 (C *
AL
− C AL )
dC AL = k L a ∫ dt
0

 C * − C AL1 
ln AL  = k at
 C* − C  L
 AL AL2 

 C * − C AL1 
Therefore a plot of ln AL 
C* − C  vs t should result in a straight line of
 AL AL 2 

slope kLa.

Static Gassing Out Method (Unsteady State)

• In the absence of respiring organism (no O2 consumption)

• Sparge vessel contents with N2, displacing O2

• Monitor variation in dissolved oxygen concentration (DO) using

a (polarographic) DO probe
 • Allow DO to fall to 0% saturation, then turn off N2 flow

• Sparge vessel contents with air at a known flowrate

• Monitor and record variation of DO concentration with respect

to time.

 C * − C AL1 
• plot of ln AL 
C* − C  vs t should result in a straight line of slope
 AL AL 2 

kLa

Dynamic Gassing Out Method

• In the presence of respiring organsims

• At time t0, turn off air supply to vessel

• Monitor and record reduction in DO between t0 and t1

• At t1, turn the air supply on again

• Monitor and record rise in DO

Theory:

For respiring systems

dC AL
dt
( )
= k L a C *AL − C AL − q o 2 X

Where X is the biomass concentration and qo2 is the specific oxygen

consumption rate (mmols O2 / g biomass s)
We assume that when we conduct dynamic kla determinations, that
the timescale of the experiment is several orders of magnitude lower
than that of the fermentation.
In this case, we can assume that a quasi-steady state exists at the
time of the experiment between oxygen transfer and oxygen
consumption.

i.e.

( −
k L a C *AL − C AL )
= qo 2 X

−
C ALis the quasi steady state oxygen concentration at the time of the
experiment

Substituting for qo2 X in the original equation

dC AL
dt
( ) ( −
= k L a C *AL − C AL − k L a C *AL − C AL )
Rearranging results in the following equation

dC AL
dt
(
= k L a C −AL − C AL )
Integrating, results in….

 C − − C AL1 
ln AL  = k a(t 2 − t1)
C− − C  L
 AL AL2 

Notes of the Dynamic Gassing Out Method

• Non-invasive method, suitable for use with respiring systems

• Assumes rapid disengagement of air bubbles when air supply is

turned off -inappropriate for highly viscous broths

• If headspace aeration is significant, use N2 blanket

 • Requires DO probe with fast response

• CL(1) determined by critical DO concentration for organisms for

bacteria and yeast Ccritical ~ 5-10% saturation

mould cultures Ccritical ~ 10-50% saturation
plant cultures Ccritical ~ 10-30% saturation

Important: On the degassing stage of the experiment,

dC AL
= −q o 2 X
dt

Therefore from the slope of the line in this region (if both axes of the
graph are in the correct units), the specific oxygen consumption rate
can be calculated once the biomass concentration is known (easy to
evaluate).

For the mathematically challenged, the slope of the line in a plot of

dC AL
CAL vs. time is !
dt
Doran 9.5 – Dynamic kLa measurement

An experiment was performed on an exponential phase microbial

culture, where the oxygen supply was disconnected and the DO
concentration was allowed to fall to 43.5% saturation. At this point,
aeration was resumed and the increase in DO concentration was
monitored with respect to time. From the following data of the
reoxygenation stage, calculate the gas-liquid mass transfer coefficient
for the reactor.

Time (s) % Saturation

10 43.5
20 53.5
30 60
40 67.5
50 70.5
60 72
70 73
100 73.5
130 73.5
 80

70

60

% Saturation
50

40

30

20

10

0
0 50 100 150
Time (s)

Figure 1. Plot of % saturation vs. time

Solution:

Whats the quasi-steady state O2 concentration?

73.5% sat.

Let 10s = t1

Plot

 C − − C AL1 
ln AL vs(t 2 − t1)
C− − C 
 AL AL2 
 4.5
4
3.5
3
2.5
ln... 2
y = 0.0611x
1.5 2
R = 0.9502
1
0.5
0
0 20 40 60 80
(t2-t1) (s)

Slope is equal to kLa – 0.0611 s-1

Note: due to the form of the equation the line has to intercept at
the origin.