1

Bioprocess Engineering I
Practical Course Manual Static KLa estimation
compiled by Dr. Sonja Diercks-Horn Fall 2011

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Introduction
The aim of this practical is to determine the oxygen mass transfer coefficient,
KLa, by static gassing out method. This method is carried out in the absence of
respiring organisms, thus there is no oxygen consumption.
Simple mass balance considerations for the concentration of dissolved oxygen giv
e:
= KLa (C*AL CAL) qO2x
(1)
where C C* AL t KLa qO2 x
= = = = = = =
oxygen concentration O2 concentration at saturation from air, in liquid time mas
s transfer coefficient specific oxygen uptake rate biomass concentration
Since the static KLa estimation is carried out in the absence of organisms equat
ion (1) is reduce to the following expression:
= KLa (C*AL CAL)
(2)
Equation (2) shows only the variation of CAL over time, the integration between
the limits t = 0 and CAL = 0 gives the following equation:
-KLa t= ln

(3)
The value C*AL for oxygen at room temperature is 10 mg L 1 (0.3125 mM) or in our
case 100 %, so equation (3) is a slope of ln(C*AL
CAL) vs t, with a slope val
ue = KLa. The experimental data to evaluate KLa must derive from different CAL
vs t values. This can be performed in a batch reactor, in which the dissolved o
xygen has been eliminated prior to any measurements by bubbling nitrogen until C
AL = 0.

3 For example: Raw data withdrawn from Erazo et al 2001

Variation of oxygen at a spirrer speed of 500 RPM
0.35 0.3 CAL (mMO2/L) 0.25 0.2 0.15 0.1 0.05 0 0 20 40 60 80 100 120 140 160 180
200 220 240 260 280 time (sec)
Fig. 1. Variation of oxygen measure in mM O2 per Litre at 500 RPM stirrer speed
The ln(C*AL CAL) was calculated and plotted against the time for the exponenti
al increase of the measured values. The linear regression was added as well as t
he formula. The KLa for this data is 41.76 h 1 calculated from the slope.
0.000 0.500 1.000 ln(C*AL CAL) 1.500 2.000 2.500 3.000 3.500 4.000 500
rpm time (sec) Linear (500 rpm) y = 0.0116x 1.1372 R = 0.9971 0 50 100 150 200
250

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Materials and Methods
Materials Fermenter with 6 L volume (20C) Carboxymethyl cellulose (Merck, 217274)
, 2110 mPas Stopwatch
Done by course instructor: Fill the vessel with 6 L water through one of the por
ts on the vessel lid Calibrate the PO2
Experiment 1 different oxygen rates
1. 2. 3. 4. 5. Set the spirrer speed to 500 rpm Sparge vessel contents with N2,
displacing O2 Monitor variation in dissolved oxygen concentration using pO2 elec
trode Allow the dissolve oxygen to fall to 0% saturation, then turn off N2 flow
Sparge vessel contents with different air ratios either at 0.2 VVM, 1 VVM or 1.5
VVM. Stopwatches must be started at the same time as when air flow starts. 6. M
onitor and record variation of dissolved oxygen concentration with respect to ti
me every 10 seconds until the same value is recorded for a span of 60 seconds. 7
. Stop the fermenter run and finally remove the water from the fermenter using t
he sample valve

5 Table 1. different oxygen rates: 0.2 VVM

seconds 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160
PO2
seconds 170 180 190 200 210 220 230 240 250 260 270 280 290 300 310 320 330
PO2
seconds 340 350 360 370 380 390 400 410 420 430 440 450 460 470 480 490 500
PO2
Table 2. different oxygen rates: 1 VVM
seconds 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160
PO2
seconds 170 180 190 200 210 220 230 240 250 260 270 280 290 300 310 320 330
PO2
seconds 340 350 360 370 380 390 400 410 420 430 440 450 460 470 480 490 500
PO2

6 Table 3. different oxygen rates: 1.5 VVM

seconds 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160
PO2
seconds 170 180 190 200 210 220 230 240 250 260 270 280 290 300 310 320 330
PO2
seconds 340 350 360 370 380 390 400 410 420 430 440 450 460 470 480 490 500
PO2
Experiment 2 different stirrer speeds
1. 2. 3. 4. 5. Set the spirrer speed either to (1) 300 rpm, (2) 500 rpm (done be
fore) or (3) 800 rpm Sparge vessel contents with N2, displacing O2 Monitor varia
tion in dissolved oxygen concentration using pO2 electrode Allow the dissolve ox
ygen to fall to 0% saturation, then turn off N2 flow Sparge vessel contents with
air at 1 VVM (liters of O2 per liter of medium per minute). Stopwatches must be
started at the same time as when air flow starts. 6. Monitor and record variati
on of dissolved oxygen concentration with respect to time every 10 seconds until
the same value is recorded for a span of 60 seconds.

7 Table 4. different spirrer speeds: 300 RPM

seconds 0 10 20 30 40 50 60 70 80 90 100 110 120 130
PO2
seconds 140 150 160 170 180 190 200 210 220 230 240 250 260 270
PO2
seconds 280 290 300 310 320 330 340 350 360 370 380 390 400 410
PO2
Table 5. different spirrer speeds: 800 RPM
seconds 0 10 20 30 40 50 60 70 80 90 100 110 120 130
PO2
seconds 140 150 160 170 180 190 200 210 220 230 240 250 260 270
PO2
seconds 280 290 300 310 320 330 340 350 360 370 380 390 400 410
PO2
Experiment 3 different viscosity of media using sodium carboxymethyl cellulose (
CMC)
1. 2. 3. 4. Fill the fermenter with 6 L of medium 1 (0.25 % CMC) using a port on
the vessel lid and a funnel Set the spirrer speed to 500 rpm Sparge vessel cont
ents with N2, displacing O2 Monitor variation in dissolved oxygen concentration
using pO2 electrode

8 5. Allow the dissolve oxygen to fall to 0% saturation, then turn off N2 flow 6
. Sparge vessel contents with air at 1 VVM. Stopwatches must be started at the s
ame time as when air flow starts. 7. Monitor and record variation of dissolved o
xygen concentration with respect to time every 10 seconds until the same value i
s recorded for a span of 60 seconds. Table 6. different oxygen rates: 0.25% CMC
seconds 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200
210 220 230 240 250 260 270
8. 9. 10. 11. 12.
PO2
seconds 280 290 300 310 320 330 340 350 360 370 380 390 400 410 420 430 440 450
460 470 480 490 500 510 520 530 540 550
PO2
seconds 560 570 580 590 600 610 620 630 640 650 660 670 680 690 700 710 720 730
740 750 760 770 780 790 800 810 820 830
PO2
Stop the fermenter run and remove medium 1 from the fermenter using the sample v
alve Fill the fermenter with 6 L of medium 2 (0.5 % CMC) using a port on the ves
sel lid and a funnel Set the spirrer speed to 500 rpm Sparge vessel contents wit
h N2, displacing O2 Monitor variation in dissolved oxygen concentration using pO
2 electrode

9 13. Allow the dissolve oxygen to fall to 0% saturation, then turn off N2 flow
14. Sparge vessel contents with air at 1 VVM. Stopwatches must be started at the
same time as when air flow starts. 15. Monitor and record variation of dissolve
d oxygen concentration with respect to time every 10 seconds until the same valu
e is recorded for a span of 60 seconds. Table 7. different oxygen rates: 0.5% CM
C
seconds 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200
210 220 230 240 250 260 270
PO2
seconds 280 290 300 310 320 330 340 350 360 370 380 390 400 410 420 430 440 450
460 470 480 490 500 510 520 530 540 550
PO2
seconds 560 570 580 590 600 610 620 630 640 650 660 670 680 690 700 710 720 730
740 750 760 770 780 790 800 810 820 830
PO2
16. Stop the fermenter run and remove medium 1 from the fermenter using the samp
le valve and clean the fermenter with water.

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References
Erazo, R. E.; Crdenas J. L. R. (2001): Determinacin experimental del coeficiente d
e transferencia de oxgeno (KLa) en un bioreactor batch. Rev. Per. Qum. Ing. Qum., V
ol. 4 (2), 22 27
Lab Report Writing Guidelines
Every student has to submit one report for the experiments. The main purpose of
lab report writing is to communicate the results to others and to enable others
to duplicate the work in a straightforward manner. A report should be as short a
s possible but contain all essential information. The lab report should be organ
ized like a scientific publication which contains the following subjects:
1. Title: This page includes a short descriptive title, the name of the person(s
) submitting the report, date of the lab course and the name of the students in
the group, the date the report is submitted, the name of the instructor. This pa
ge is not numbered. 2. Table of Content: The Table of Contents provides page loc
ations of major sections. 3. Introduction: Explains its objectives, significance
, and provides the background necessary to understand the experiment. When appro
priate, the background should indicate theoretical prediction. This section is n
ot intended to be a simple reproduction of some texts (do not forget to cite you
r references in the text, see how to cite below); instead it must reflect your u
nderstanding about information pertaining to the experiment, and must emphasize
the importance and applications of the experimental subject. Direct reproduction
s from earlier reports, books, and/or internet will be considered as cheating an
d be subjected to a penalty. 4. Materials and methods: if the procedure is the s
ame as in the manual, cite the manual. Write only the changes you made into the
lab report. 5. Results: present all data obtained and calculated, graphs and des
cription of data, e.g. SDS gels. When presenting graphs, make sure that your spe
cific measurement data points are indicated on the graphs. Do not just show a li
ne or a curve. Sufficiently detailed explanations should precede each table and/
or graph in regard to the experimental conditions, range of parameters, so as to
allow the reader to follow and understand the meaning of the information presen
ted. Each table or graph has to have a caption (Figures have legends while Table
s have headings!), proper labels and subheadings, names of parameters (with unit
s) used in axes of graphs or in the table column headings. The symbols used must
be the same as those used in the text. The results section further includes any
unexpected observations that you made during the experiment (they could help to
explain unexpected outcomes in the discussion). If you made a mistake do not tr
y to cover it up but describe it. Do not yet interpret or discuss your data, thi
s belongs to the discussion! 6. Discussion: This section places specific results
into the context of the experiment as a whole. Analyze the results and discuss
their implications, compare experimental results and

11 expected, acknowledge possible sources of error. What would you make better i
f you have to do the experiment again. 7. Reference: Good scientific practice in
cludes that all information taken from other sources (textbooks, articles, Lab M
anuals, Internet) needs to be indicated as such. In addition, the different sour
ces of information have to be cited in the correct way. The correct way means th
e way citations are placed in scientific articles. Every listed reference must b
e cited in the text of the report by author and year (for example, Bird et al.,
1960). In the last part of the lab report, you place the list of citations in al
phabetical order based on first author and you write out all authors, year of pu
blishing, title, Journal or Publisher. The internet sources are given at the end
.
Examples for a reference list: Abramowitz, M., Stegun, I.A., (1965): Handbook of
Mathematical Functions. Dover, New York Winslow, F.H., and Matreyek, W., (1951)
: Pyrolysis of Crosslinked Styrene Polymers. Journal of Polymer Science, 22, pp.
315 324.
Tasks for the lab report
Plot the measurement of the three different experiments (RPM, VVM, viscosity) in
three different plots with the PO2 values on the y axis and the time on the x a
xis Calculate the ln(C*AL CAL) and plot the three different experiments (RPM,
VVM, viscosity) in three different plots with ln(C*AL
CAL) on the y axis and t
he time on the x axis Calculate the KLa from the slope and present the values in
a table. Discuss your results

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MATERIAL SAFETY DATA SHEET
Name (chemical and trivial) Chemical formula Structural formula Sodium carboxyme
thyl cellulose
Molar mass/Molecular weight Density General classification Functional groups
GHS Pictograms H statements P statements Signal word Gloves Safety goggles Hood
Role in the experiment
Additional notes
_________________________________ Printed Name and Signature