1.

0 ABSTRACT

The aim of this experiment to measure the volumetric mass transfer coefficient (kLa) of
a stirred tank reactor with bubble aeration at different temperature, aeration and agitation. This
experiment was conducted by using a bioreactor. The method used to determining the kLa value
is called the unsteady-state method because the absent of cell. Nitrogen gas was purged into
the system until the value of oxygen became 0%. The calibration was done before the reading
of dissolved oxygen (DO %) was taken. The bioreactor’s monitor was made sure that it stopped
blinking to indicate the calibration was done correctly even though the value becomes negative.
When the value became negative, the value was adjusted manually until it became zero. The
bioreactor was aerated with air for 100%. The reading was taken for every 5 seconds until the
increasing DO % reaching three stable values. To investigate the effects of different
temperatures on the kLa value, the bioreactor’s parameter such as aeration and agitation were
set fixed at 2 L/m and 400 rpm respectively. On the other hand, to investigate the effects of
different aeration on the kLa value, the bioreactor’s parameter such as temperature and
agitation were set fixed at 30°C and 400 rpm respectively. Lastly, to investigate the effects of
different agitation on the kLa value, the bioreactor’s parameter such as temperature and
aeration were set fixed at 30°C and 2 L/min respectively. Based on the unsteady state method,
the graph of ln(C*-CL) versus time was plotted to estimate the value of kLa for each
manipulated variables. Next, the kLa values obtained for each temperature, aeration and
agitation were tabulated and compared by plotting graph of kLa against its manipulated
variables. For the temperature at 35°C, 40°C, 45°C and 50°C, the kLa value obtained are 148.68
h-1, 182.52 h-1, 219.24 h-1 and 236.52 h-1. For the aeration rate at 0.5, 1.0, 1.5, 2.0 and 2.5 L/min,
the kLa obtained is 36.72 h-1, 45.36 h-1, 73.44 h-1, 95.76 h-1 and 115.20 h-1 respectively. For the
agitation speed at 200 rpm, 400 rpm, 600 rpm, 800 rpm and 1000 rpm, the k La value obtained
is 51.48 h-1, 99.36 h-1, 154.08 h-1, 185.04 h-1 and 223.2 h-1 respectively. The aims of the
experiment is achieved. Hence, the experiment was conducted successfully.
2.0 INTRODUCTION

Majority of the biochemical processes require oxygen as the source to produce the
product. Fermentation is one of the examples of the processes. The organisms needs oxygen to
initiate the reaction and yield the desired product. Hence, dissolved oxygen concentration
becomes one of the important control variables in any aerobic fermentation.

Three basic reactor types for the aerobic cultivation of suspended cells are systems with
internal mechanical agitation, bubble columns, and loop reactors (Shuler & Kargi, 2002). The
traditional fermenter is the stirred-tank reactor (Figure 2.1), the prime example of a reactor with
internal mechanical agitation (Shuler & Kargi, 2002). The main virtues of such systems are
that they are highly flexible and can provide high kLa (volumetric mass-transfer coefficient)
values for gas transfer (Shuler & Kargi, 2002). Mass transfer coefficient, kLa is a function of
the mechanics of a particular reactor namely its constant dimension and its operating
parameters. Figure 2.1 below shows the design of a stirred-tank reactor.

Figure 2.1: Stirred-tank reactor (Shuler & Kargi, 2002)

It is very important to thoroughly understand the oxygen transfer to cells in a reactor.

During aerobic fermentation, oxygen has to be supplied continuously to the reaction liquid in
order to maintain the design concentration while oxygen is consumed by the organism. The
oxygen is transferred from air bubbles and then sparged into the reaction solution and broken
up and mixed by mechanical stirring.
The kLa values are dependent on factors such as agitation rate, gas velocity, bubble size,
temperature, gas sparging system and cell morphology (Sathish & Amaranth, 2015, 7(3)). The
determination of kLa in a bioreactor is essential in order to establish its aeration efficiency and
to quantify the effects of operating variables on oxygen supply (Moutafchieva, Popova,
Dimitrova, & Tchaoushev, 2013, 48(4)). Oxygen transfer in aerobic bioprocesses is essential
and any shortage of oxygen vastly affects the process performance (Moutafchieva, Popova,
Dimitrova, & Tchaoushev, 2013, 48(4)). Therefore, oxygen mass transfer is one of the most
important phenomena in the design and operation of mixing-sparging equipment for
bioreactors (Moutafchieva, Popova, Dimitrova, & Tchaoushev, 2013, 48(4)).
Although kLa is difficult to predict, it is a measurable parameter. Four approaches are
commonly used: unsteady state, steady state, dynamic, and sulphide test (Shuler & Kargi, 2002).
3.0 AIMS

The aim of this experiment is:

1. To measure the volumetric mass transfer coefficient (kLa) of a stirred tank reactor with
bubble aeration.
2. To study the relationship between temperature, aeration and agitation between the
volumetric mass transfer coefficient (kLa).
4.0 THEORY

For industrial-scale fermenters, oxygen supply and heat removal are the key design
limitations (Shuler & Kargi, 2002). Although kLa is difficult to predict, it is a measurable
parameter (Shuler & Kargi, 2002). Four approaches are commonly used: unsteady state, steady
state, dynamic, and sulphide test (Shuler & Kargi, 2002). The way in which these methods are
applied depends on whether the test is being made on the system in the presence or absence of
cells (Shuler & Kargi, 2002).

In this experiment, the unsteady-state method, which is also called the static gassing
out method is used to determine the mass transfer coefficient (kLa). By using this method, the
oxygen transfer rate, OTR, is involved but the oxygen uptake rate, OUR, is not included due
to the absence of cells. A new reactor prior to operation can be filled with pure water or a
medium in which C* can be accurately measured. Oxygen is removed from the system by
sparging with N2 (Shuler & Kargi, 2002). With the unsteady-state method, air is then introduced
and the change in dissolved oxygen (DO) is monitored until the solution is nearly saturated.
When oxygen uptake rate, OUR = 0, the oxygen mass balance in the liquid phase can be
simplified to equation below (Moutafchieva, Popova, Dimitrova, & Tchaoushev, 2013, 48(4)).

𝑑𝐶𝐿
= 𝑘𝐿 𝑎(𝐶 ∗ − 𝐶𝐿 ) = 𝑂𝑇𝑅
𝑑𝑡

−𝑑(𝐶 ∗ − 𝐶𝐿 )
= 𝑘𝐿 𝑎
𝑑𝑡/(𝐶 ∗ − 𝐶𝐿 )

ln(𝐶 ∗ − 𝐶𝐿 ) = − 𝑘𝐿 𝑎𝑡

Where,

kL = oxygen transfer coefficient (cm/h)

a = gas-liquid interfacial area (cm2/cm3)

kLa = mass transfer coefficient (h-1)

C* = saturated dissolved oxygen concentration (mg/L)

CL = actual dissolved oxygen concentration (mg/L)

qO2 = specific rate of oxygen consumption (mmol O2/gdw.h)

X = bacteria concentration (gdw/L)

gdw = gram dry weight

OTR = oxygen transfer rate

OUR = oxygen uptake rate

Hence, a plot of ln (C*˗ CL) versus time, t, should result in a straight line of slope –kLa
(Moutafchieva, Popova, Dimitrova, & Tchaoushev, 2013, 48(4)). As shown in Figure 4.2, a
plot of log (C* - CL) versus time will give an estimate of kLa (Shuler & Kargi, 2002).

Figure 4.2: Plot of log (C*˗ CL) versus time (Shuler & Kargi, 2002)
5.0 APPARATUS

1. Bioreactor
2. Distilled water
3. Stopwatch
4. Oxygen
5. Nitrogen gas
6. HI-BLOW (HP 80) linear air pump aerator

Pump
Reactor

Bioreactor
monitor

Figure 5.1 Bioreactor

6.0 PROCEDURE

6.1 Procedures for temperature as manipulated variable

1. PO2 probe was polarized for two hours before the main experiment was started.
2. The bioreactor’s parameter such as aeration and agitation were fixed which were at 2
L/min and 400 rpm respectively.
3. The pump was switched off.
4. Temperature was first set at 30°C and prepared for two point calibration. The setting
was done before purging the nitrogen.
5. Nitrogen gas was purged into the system until the value of oxygen became 0%.
6. The bioreactor’s monitor was made sure to stop blinking to indicate the calibration
was done correctly even though the value became negative.
7. When the value was negative, the value was adjusted manually until the value was 0.
8. The nitrogen valve was closed after the value was 0 and the nitrogen wire was
disconnected from the bioreactor.
9. Pump was switched on.
10. The bioreactor was aerated with air for 100%.
11. Timer was set and reading was taken for every 5 seconds until the increasing of DO%
achieved three stable values.
12. The results were recorded and a graph of DO% versus time was plotted.
13. Step 5-12 were repeated using different temperature, which is 35°C, 40°C, 45°C and
50°C.

6.2 Procedures for aeration as manipulated variable

1. PO2 probe was polarized for two hours before the main experiment was started.
2. The bioreactor’s parameter such as temperature and agitation were fixed which were
at 30°C and 400 rpm respectively.
3. The pump was switched off.
4. Aeration was first set at 0.5 L/min and prepared for two point calibration. The setting
was done before purging the nitrogen.
5. Nitrogen gas was purged into the system until the value of oxygen became 0%.
6. The bioreactor’s monitor was made sure to stop blinking to indicate the calibration
was done correctly even though the value became negative.
7. When the value was negative, the value was adjusted manually until the value was 0.
 8. The nitrogen valve was closed after the value was 0 and the nitrogen wire was
disconnected from the bioreactor.
9. Pump was switched on.
10. The bioreactor was aerated with air for 100%.
11. Timer was set and reading was taken for every 5 seconds until the increasing of DO%
achieved three stable values.
12. The results were recorded and a graph of DO% versus time was plotted.
13. Step 5-12 were repeated using different aeration, which is 1 L/min, 1.5 L/min, 2.0
L/min and 2.5 L/min.

6.3 Procedures for agitation as manipulated variable

1. PO2 probe was polarized for two hours before the main experiment was started.
2. The bioreactor’s parameter such as aeration and temperature were fixed which were
at 2 L/min and 30°C respectively.
3. The pump was switched off.
4. Agitation was first set at 200 rpm and prepared for two point calibration. The setting
was done before purging the nitrogen.
5. Nitrogen gas was purged into the system until the value of oxygen became 0%.
6. The bioreactor’s monitor was made sure to stop blinking to indicate the calibration
was done correctly even though the value became negative.
7. When the value was negative, the value was adjusted manually until the value was 0.
8. The nitrogen valve was closed after the value was 0 and the nitrogen wire was
disconnected from the bioreactor.
9. Pump was switched on.
10. The bioreactor was aerated with air for 100%.
11. Timer was set and reading was taken for every 5 seconds until the increasing of DO%
achieved three stable values.
12. The results were recorded and a graph of DO% versus time was plotted.
13. Step 5-12 were repeated using different agitation, which is 400 rpm, 600 rpm, 800
rpm, and 1000 rpm.
7.0 RESULTS

7.1 Results for temperature as manipulated variable

Constant variables:
Aeration = 2 L/min
Agitation = 400 rpm

Time (s) Temperature 35ºC (C* = 7 mg/L)

PO2% CL C*-CL ln(C*-CL)
5 0.20 0.0140 6.9860 1.9439
10 3.78 0.2646 6.7354 1.9074
15 11.30 0.7910 6.2090 1.8260
20 20.50 1.4350 5.5650 1.7165
25 30.40 2.1280 4.8720 1.5835
30 41.40 2.8980 4.1020 1.4115
35 48.10 3.3670 3.6330 1.2901
40 55.60 3.8920 3.1080 1.1340
45 61.90 4.3330 2.6670 0.9810
50 68.90 4.8230 2.1770 0.7779
55 74.40 5.2080 1.7920 0.5833
60 78.50 5.4950 1.5050 0.4088
65 82.20 5.7540 1.2460 0.2199
70 85.40 5.9780 1.0220 0.0218
75 87.70 6.1390 0.8610 -0.1497
80 90.00 6.3000 0.7000 -0.3567
85 91.80 6.4260 0.5740 -0.5551
90 93.40 6.5380 0.4620 -0.7722
95 94.70 6.6290 0.3710 -0.9916
100 95.80 6.7060 0.2940 -1.2242
105 96.60 6.7620 0.2380 -1.4355
110 97.40 6.8180 0.1820 -1.7037
115 98.00 6.8600 0.1400 -1.9661
120 98.70 6.9090 0.0910 -2.3969
125 99.00 6.9300 0.0700 -2.6593
130 99.40 6.9580 0.0420 -3.1701
135 99.70 6.9790 0.0210 -3.8632
140 100.00 7.0000 0.0000 -
145 100.00 7.0000 0.0000 -
150 100.00 7.0000 0.0000 -
155 100.00 7.0000 0.0000 -
160 100.00 7.0000 0.0000 -

Table 7.1.1: Table for temperature at 35ºC at each 5 seconds

Graph 7.1.1: Graph of ln (C*-CL) versus time for temperature 35°C

Time (s) Temperature 40ºC (C* = 6.5 mg/L)

PO2% CL C*-CL ln(C*-CL)
5 0.00 0.0000 6.5000 1.8718
10 2.85 0.1853 6.3148 1.8429
15 13.40 0.8710 5.6290 1.7279
20 24.60 1.5990 4.9010 1.5894
25 34.90 2.2685 4.2315 1.4426
30 44.30 2.8795 3.6205 1.2866
35 52.10 3.3865 3.1135 1.1357
40 60.90 3.9585 2.5415 0.9328
45 67.90 4.4135 2.0865 0.7355
50 73.30 4.7645 1.7355 0.5513
55 78.30 5.0895 1.4105 0.3439
60 82.50 5.3625 1.1375 0.1288
65 85.90 5.5835 0.9165 -0.0872
70 88.90 5.7785 0.7215 -0.3264
75 91.60 5.9540 0.5460 -0.6051
80 93.70 6.0905 0.4095 -0.8928
85 95.50 6.2075 0.2925 -1.2293
90 96.80 6.2920 0.2080 -1.5702
95 98.00 6.3700 0.1300 -2.0402
100 99.00 6.4350 0.0650 -2.7334
105 99.80 6.4870 0.0130 -4.3428
110 100.00 6.5000 0.0000 -

Table 7.1.2: Table for temperature at 40ºC at each 5 seconds

 Graph 7.1.2: Graph of ln (C*-CL) versus time for temperature 40°C

Time (s) Temperature 45ºC (C* = 6 mg/L)

PO2% CL C*-CL ln(C*-CL)
5 0 0.0000 6.0000 1.7918
10 7.93 0.4758 5.5242 1.7091
15 22.90 1.3740 4.6260 1.5317
20 36.00 2.1600 3.8400 1.3455
25 50.40 3.0240 2.9760 1.0906
30 61.70 3.7020 2.2980 0.8320
35 69.20 4.1520 1.8480 0.6141
40 77.00 4.6200 1.3800 0.3221
45 81.80 4.9080 1.0920 0.0880
50 87.10 5.2260 0.7740 -0.2562
55 90.90 5.4540 0.5460 -0.6051
60 94.30 5.6580 0.3420 -1.0729
65 96.40 5.7840 0.2160 -1.5325
 70 98.70 5.9220 0.0780 -2.5510
75 100.00 6.0000 0.0000 -

Table 7.1.3: Table for temperature at 45ºC at each 5 seconds

Graph 7.1.3: Graph of ln (C*-CL) versus time for temperature 45°C

Time (s) Temperature 50ºC (C* = 5.6 mg/L)

PO2% CL C*-CL ln(C*-CL)
5 0 0.0000 5.6000 1.7228
10 5.07 0.2839 5.3161 1.6707
15 18.70 1.0472 4.5528 1.5157
20 28.10 1.5736 4.0264 1.3929
25 37.60 2.1056 3.4944 1.2512
30 51.90 2.9064 2.6936 0.9909
35 62.00 3.4720 2.1280 0.7552
40 70.50 3.9480 1.6520 0.5020
45 78.00 4.3680 1.2320 0.2086
50 83.20 4.6592 0.9408 -0.0610
55 89.00 4.9840 0.6160 -0.4845
60 93.00 5.2080 0.3920 -0.9365
65 96.50 5.4040 0.1960 -1.6296
70 99.50 5.5720 0.0280 -3.5756

Table 7.l.4: Table for temperature at 50ºC at each 5 seconds

Graph 7.1.4: Graph of ln (C*-CL) versus time for temperature 50°C

Temperature, T (°C) kLa (h-1)

35 148.68
40 182.52
45 219.24
50 236.52

Table 7.1.5: Table of kLa value for temperature at 35, 40, 45 and 50 °C
 Graph 7.1.5: Graph of kLa versus temperature

7.2 Results for aeration as manipulated variable

Constant variables:
Temperature = 30°C
C* = 7.6 mg/L
Agitation = 400 rpm

Time (s) Aeration = 0.5 L/min

PO2% CL C*-CL ln(C*-CL)

5 9.95 0.7562 6.8438 1.9233
10 15.20 1.1552 6.4448 1.8633
15 20.20 1.5352 6.0648 1.8025
20 25.10 1.9076 5.6924 1.7391
25 30.10 2.2876 5.3124 1.6700
30 34.30 2.6068 4.9932 1.6081
35 38.80 2.9488 4.6512 1.5371
40 42.50 3.2300 4.3700 1.4748
45 46.10 3.5036 4.0964 1.4101
50 49.00 3.7240 3.8760 1.3548
55 52.20 3.9672 3.6328 1.2900
60 55.10 4.1876 3.4124 1.2274
65 57.90 4.4004 3.1996 1.1630
70 60.00 4.5600 3.0400 1.1119
75 62.50 4.7500 2.8500 1.0473
80 64.40 4.8944 2.7056 0.9953
85 66.20 5.0312 2.5688 0.9434
90 68.00 5.1680 2.4320 0.8887
95 70.10 5.3276 2.2724 0.8208
100 72.00 5.4720 2.1280 0.7552
105 73.50 5.5860 2.0140 0.7001
110 74.90 5.6924 1.9076 0.6458
115 76.20 5.7912 1.8088 0.5927
120 77.50 5.8900 1.7100 0.5365
125 78.70 5.9812 1.6188 0.4817
130 79.80 6.0648 1.5352 0.4287
135 81.00 6.1560 1.4440 0.3674
140 82.00 6.2320 1.3680 0.3133
145 82.90 6.3004 1.2996 0.2621
150 83.80 6.3688 1.2312 0.2080
155 84.70 6.4372 1.1628 0.1508
160 85.60 6.5056 1.0944 0.0902
165 86.10 6.5436 1.0564 0.0549
170 86.80 6.5968 1.0032 0.0032
175 87.50 6.6500 0.9500 -0.0513
180 88.00 6.6880 0.9120 -0.0921
185 88.70 6.7412 0.8588 -0.1522
190 89.10 6.7716 0.8284 -0.1883
195 89.90 6.8324 0.7676 -0.2645
200 90.20 6.8552 0.7448 -0.2946
205 90.70 6.8932 0.7068 -0.3470
210 91.00 6.9160 0.6840 -0.3798
215 91.50 6.9540 0.6460 -0.4370
220 91.90 6.9844 0.6156 -0.4852
225 92.50 7.0300 0.5700 -0.5621
230 92.80 7.0528 0.5472 -0.6029
235 93.00 7.0680 0.5320 -0.6311
240 93.30 7.0908 0.5092 -0.6749
245 93.70 7.1212 0.4788 -0.7365
250 93.90 7.1364 0.4636 -0.7687
255 94.30 7.1668 0.4332 -0.8366
260 94.50 7.1820 0.4180 -0.8723
265 94.80 7.2048 0.3952 -0.9284
270 95.00 7.2200 0.3800 -0.9676
275 95.10 7.2276 0.3724 -0.9878
280 95.30 7.2428 0.3572 -1.0295
285 95.50 7.2580 0.3420 -1.0729
290 95.90 7.2884 0.3116 -1.1660
295 96.00 7.2960 0.3040 -1.1907
300 96.10 7.3036 0.2964 -1.2160
305 96.30 7.3188 0.2812 -1.2687
310 96.50 7.3340 0.2660 -1.3243
315 96.60 7.3416 0.2584 -1.3532
320 96.80 7.3568 0.2432 -1.4139
325 97.00 7.3720 0.2280 -1.4784
330 97.10 7.3796 0.2204 -1.5123
335 97.20 7.3872 0.2128 -1.5474
340 97.40 7.4024 0.1976 -1.6215
345 97.50 7.4100 0.1900 -1.6607
350 97.60 7.4176 0.1824 -1.7016
355 97.70 7.4252 0.1748 -1.7441
360 97.80 7.4328 0.1672 -1.7886
365 98.00 7.4480 0.1520 -1.8839
370 98.10 7.4556 0.1444 -1.9352
375 98.10 7.4556 0.1444 -1.9352
380 98.20 7.4632 0.1368 -1.9892
385 98.20 7.4632 0.1368 -1.9892
390 98.30 7.4708 0.1292 -2.0464
395 98.40 7.4784 0.1216 -2.1070
400 98.40 7.4784 0.1216 -2.1070
405 98.60 7.4936 0.1064 -2.2405
410 98.60 7.4936 0.1064 -2.2405
415 98.60 7.4936 0.1064 -2.2405
420 98.80 7.5088 0.0912 -2.3947
425 98.80 7.5088 0.0912 -2.3947
430 98.90 7.5164 0.0836 -2.4817
435 99.00 7.5240 0.0760 -2.5770
440 99.00 7.5240 0.0760 -2.5770
445 99.00 7.5240 0.0760 -2.5770
450 99.20 7.5392 0.0608 -2.8002
455 99.30 7.5468 0.0532 -2.9337
460 99.30 7.5468 0.0532 -2.9337
465 99.30 7.5468 0.0532 -2.9337
470 99.30 7.5468 0.0532 -2.9337
475 99.40 7.5544 0.0456 -3.0878
480 99.50 7.5620 0.0380 -3.2702
485 99.50 7.5620 0.0380 -3.2702
490 99.50 7.5620 0.0380 -3.2702
495 99.50 7.5620 0.0380 -3.2702
500 99.50 7.5620 0.0380 -3.2702

Table 7.2.1: Table for aeration 0.5 L/min at each 5 seconds

 Graph 7.2.1: Graph of ln (C*-CL) versus time for aeration 0.5 L/min

Time (s) Aeration = 1.0 L/min

PO2% CL C*-CL ln(C*-CL)
5 0.05 0.0038 7.5962 2.0276
10 3.24 0.2462 7.3538 1.9952
15 5.59 0.4248 7.1752 1.9706
20 8.66 0.6582 6.9418 1.9376
25 12.5 0.9500 6.6500 1.8946
30 17.1 1.2996 6.3004 1.8406
35 22.1 1.6796 5.9204 1.7784
40 26.6 2.0216 5.5784 1.7189
45 30.8 2.3408 5.2592 1.6600
50 35.1 2.6676 4.9324 1.5958
55 39.2 2.9792 4.6208 1.5306
60 43.1 3.2756 4.3244 1.4643
65 46.7 3.5492 4.0508 1.3989
70 50.7 3.8532 3.7468 1.3209
75 53.8 4.0888 3.5112 1.2560
80 56.5 4.2940 3.3060 1.1957
85 59.8 4.5448 3.0552 1.1168
90 62.4 4.7424 2.8576 1.0500
95 65.0 4.9400 2.6600 0.9783
100 67.4 5.1224 2.4776 0.9073
105 69.6 5.2896 2.3104 0.8374
110 71.8 5.4568 2.1432 0.7623
115 73.7 5.6012 1.9988 0.6925
120 75.9 5.7684 1.8316 0.6052
125 77.4 5.8824 1.7176 0.5409
130 78.1 5.9356 1.6644 0.5095
135 80.4 6.1104 1.4896 0.3985
140 81.7 6.2092 1.3908 0.3299
145 83.0 6.3080 1.2920 0.2562
150 84.1 6.3916 1.2084 0.1893
155 85.1 6.4676 1.1324 0.1243
160 86.0 6.5360 1.0640 0.0620
165 86.9 6.6044 0.9956 -0.0044
170 87.9 6.6804 0.9196 -0.0838
175 88.7 6.7412 0.8588 -0.1522
180 89.3 6.7868 0.8132 -0.2068
185 90.1 6.8476 0.7524 -0.2845
190 90.7 6.8932 0.7068 -0.3470
195 91.3 6.9388 0.6612 -0.4137
200 91.8 6.9768 0.6232 -0.4729
205 92.3 7.0148 0.5852 -0.5358
210 92.8 7.0528 0.5472 -0.6029
215 93.3 7.0908 0.5092 -0.6749
220 93.6 7.1136 0.4864 -0.7207
225 94.0 7.1440 0.4560 -0.7853
230 94.4 7.1744 0.4256 -0.8543
235 94.8 7.2048 0.3952 -0.9284
240 95.1 7.2276 0.3724 -0.9878
245 95.3 7.2428 0.3572 -1.0295
250 95.6 7.2656 0.3344 -1.0954
255 95.9 7.2884 0.3116 -1.1660
260 96.1 7.3036 0.2964 -1.2160
265 96.3 7.3188 0.2812 -1.2687
270 96.5 7.3340 0.2660 -1.3243
275 96.7 7.3492 0.2508 -1.3831
280 96.9 7.3644 0.2356 -1.4456
285 97.0 7.3720 0.2280 -1.4784
290 97.1 7.3796 0.2204 -1.5123
295 97.4 7.4024 0.1976 -1.6215
300 97.5 7.4100 0.1900 -1.6607
305 97.7 7.4252 0.1748 -1.7441
310 97.8 7.4328 0.1672 -1.7886
315 98.0 7.4480 0.1520 -1.8839
320 98.1 7.4556 0.1444 -1.9352
325 98.1 7.4556 0.1444 -1.9352
330 98.2 7.4632 0.1368 -1.9892
335 98.3 7.4708 0.1292 -2.0464
340 98.4 7.4784 0.1216 -2.1070
345 98.5 7.4860 0.1140 -2.1716
350 98.6 7.4936 0.1064 -2.2405
355 98.7 7.5012 0.0988 -2.3147
360 98.9 7.5164 0.0836 -2.4817
365 98.9 7.5164 0.0836 -2.4817
370 98.9 7.5164 0.0836 -2.4817
375 99.0 7.5240 0.0760 -2.5770
380 99.1 7.5316 0.0684 -2.6824
385 99.1 7.5316 0.0684 -2.6824
 390 99.1 7.5316 0.0684 -2.6824
395 99.2 7.5392 0.0608 -2.8002
400 99.3 7.5468 0.0532 -2.9337
405 99.3 7.5468 0.0532 -2.9337
410 99.3 7.5468 0.0532 -2.9337
415 99.3 7.5468 0.0532 -2.9337
420 99.3 7.5468 0.0532 -2.9337

Table 7.2.2: Table for aeration 1.0 L/min at each 5 seconds

Graph 7.2.2: Graph of ln (C*-CL) versus time for aeration 1.0 L/min

Time (s) Aeration = 1.5 L/min

PO2% CL C*-CL ln(C*-CL)
5 3.00 0.2280 7.3720 1.9977
10 6.84 0.5198 7.0802 1.9573
15 10.00 0.7600 6.8400 1.9228
20 17.00 1.2920 6.3080 1.8418
25 22.10 1.6796 5.9204 1.7784
30 28.30 2.1508 5.4492 1.6955
35 34.10 2.5916 5.0084 1.6111
40 39.00 2.9640 4.6360 1.5339
45 44.90 3.4124 4.1876 1.4321
50 51.00 3.8760 3.7240 1.3148
55 54.80 4.1648 3.4352 1.2341
60 58.60 4.4536 3.1464 1.1463
65 62.50 4.7500 2.8500 1.0473
70 65.10 4.9476 2.6524 0.9755
75 69.10 5.2516 2.3484 0.8537
80 72.60 5.5176 2.0824 0.7335
85 74.50 5.6620 1.9380 0.6617
90 76.90 5.8444 1.7556 0.5628
95 79.10 6.0116 1.5884 0.4627
100 81.70 6.2092 1.3908 0.3299
105 84.40 6.4144 1.1856 0.1702
110 86.20 6.5512 1.0488 0.0476
115 87.90 6.6804 0.9196 -0.0838
120 89.10 6.7716 0.8284 -0.1883
125 90.10 6.8476 0.7524 -0.2845
130 91.30 6.9388 0.6612 -0.4137
135 92.10 6.9996 0.6004 -0.5102
140 92.90 7.0604 0.5396 -0.6169
145 93.50 7.1060 0.4940 -0.7052
150 94.10 7.1516 0.4484 -0.8021
155 94.50 7.1820 0.4180 -0.8723
160 95.10 7.2276 0.3724 -0.9878
165 95.60 7.2656 0.3344 -1.0954
170 96.00 7.2960 0.3040 -1.1907
175 96.30 7.3188 0.2812 -1.2687
180 96.70 7.3492 0.2508 -1.3831
185 97.10 7.3796 0.2204 -1.5123
190 97.40 7.4024 0.1976 -1.6215
195 97.60 7.4176 0.1824 -1.7016
200 97.90 7.4404 0.1596 -1.8351
205 98.10 7.4556 0.1444 -1.9352
210 98.20 7.4632 0.1368 -1.9892
215 98.40 7.4784 0.1216 -2.1070
220 98.50 7.4860 0.1140 -2.1716
225 98.70 7.5012 0.0988 -2.3147
230 98.90 7.5164 0.0836 -2.4817
235 99.00 7.5240 0.0760 -2.5770
240 99.00 7.5240 0.0760 -2.5770
245 99.20 7.5392 0.0608 -2.8002
250 99.20 7.5392 0.0608 -2.8002
255 99.30 7.5468 0.0532 -2.9337
260 99.40 7.5544 0.0456 -3.0878
265 99.40 7.5544 0.0456 -3.0878
270 99.50 7.5620 0.0380 -3.2702
275 99.50 7.5620 0.0380 -3.2702
280 99.60 7.5696 0.0304 -3.4933
285 99.60 7.5696 0.0304 -3.4933
290 99.60 7.5696 0.0304 -3.4933
295 99.70 7.5772 0.0228 -3.7810
300 99.70 7.5772 0.0228 -3.7810
305 99.70 7.5772 0.0228 -3.7810

Table 7.2.3: Table for aeration 1.5 L/min at each 5 seconds

 Graph 7.2.3: Graph of ln (C*-CL) versus time for aeration 1.5 L/min

Time (s) Aeration = 2.0 L/min

PO2% CL C*-CL ln(C*-CL)
5 0.10 0.0076 7.5924 2.0271
10 2.59 0.1968 7.4032 2.0019
15 6.69 0.5084 7.0916 1.9589
20 12.60 0.9576 6.6424 1.8935
25 19.00 1.4440 6.1560 1.8174
30 25.50 1.9380 5.6620 1.7338
35 32.30 2.4548 5.1452 1.6381
40 39.90 3.0324 4.5676 1.5190
45 45.10 3.4276 4.1724 1.4285
50 50.70 3.8532 3.7468 1.3209
55 57.40 4.3624 3.2376 1.1748
60 61.00 4.6360 2.9640 1.0865
65 65.30 4.9628 2.6372 0.9697
70 69.00 5.2440 2.3560 0.8570
75 73.00 5.5480 2.0520 0.7188
80 76.10 5.7836 1.8164 0.5969
85 78.00 5.9280 1.6720 0.5140
90 81.60 6.2016 1.3984 0.3353
95 83.60 6.3536 1.2464 0.2203
100 85.40 6.4904 1.1096 0.1040
105 87.10 6.6196 0.9804 -0.0198
110 88.80 6.7488 0.8512 -0.1611
115 90.10 6.8476 0.7524 -0.2845
120 91.50 6.9540 0.6460 -0.4370
125 92.60 7.0376 0.5624 -0.5755
130 93.40 7.0984 0.5016 -0.6900
135 94.30 7.1668 0.4332 -0.8366
140 95.00 7.2200 0.3800 -0.9676
145 95.60 7.2656 0.3344 -1.0954
150 96.20 7.3112 0.2888 -1.2420
155 96.60 7.3416 0.2584 -1.3532
160 97.00 7.3720 0.2280 -1.4784
165 97.40 7.4024 0.1976 -1.6215
170 97.80 7.4328 0.1672 -1.7886
175 98.00 7.4480 0.1520 -1.8839
180 98.20 7.4632 0.1368 -1.9892
185 98.30 7.4708 0.1292 -2.0464
190 98.50 7.4860 0.1140 -2.1716
195 98.70 7.5012 0.0988 -2.3147
200 99.00 7.5240 0.0760 -2.5770
205 99.10 7.5316 0.0684 -2.6824
210 99.20 7.5392 0.0608 -2.8002
215 99.30 7.5468 0.0532 -2.9337
220 99.40 7.5544 0.0456 -3.0878
225 99.50 7.5620 0.0380 -3.2702
 230 99.60 7.5696 0.0304 -3.4933
235 99.70 7.5772 0.0228 -3.7810
240 99.80 7.5848 0.0152 -4.1865
245 99.90 7.5924 0.0076 -4.8796
250 99.90 7.5924 0.0076 -4.8796
255 100.00 7.6000 0.0000 -

Table 7.2.4: Table for aeration 2.0 L/min at each 5 seconds

Graph 7.2.4: Graph of ln (C*-CL) versus time for aeration 2.0 L/min

Time (s) Aeration = 2.5 L/min

PO2% CL C*-CL ln(C*-CL)
5 2.040 0.1550 7.4450 2.0075
10 7.090 0.5388 7.0612 1.9546
15 14.04 1.0670 6.5330 1.8769
20 20.60 1.5656 6.0344 1.7975
25 28.20 2.1432 5.4568 1.6969
30 34.80 2.6448 4.9552 1.6004
35 42.20 3.2072 4.3928 1.4800
40 48.90 3.7164 3.8836 1.3568
45 54.80 4.1648 3.4352 1.2341
50 59.50 4.5220 3.0780 1.1243
55 64.80 4.9248 2.6752 0.9840
60 69.70 5.2972 2.3028 0.8341
65 73.40 5.5784 2.0216 0.7039
70 76.90 5.8444 1.7556 0.5628
75 79.90 6.0724 1.5276 0.4237
80 82.60 6.2776 1.3224 0.2794
85 85.00 6.4600 1.1400 0.1310
90 87.00 6.6120 0.9880 -0.0121
95 88.80 6.7488 0.8512 -0.1611
100 90.20 6.8552 0.7448 -0.2946
105 91.20 6.9312 0.6688 -0.4023
110 92.90 7.0604 0.5396 -0.6169
115 93.80 7.1288 0.4712 -0.7525
120 94.70 7.1972 0.4028 -0.9093
125 95.50 7.2580 0.3420 -1.0729
130 96.10 7.3036 0.2964 -1.2160
135 96.70 7.3492 0.2508 -1.3831
140 97.10 7.3796 0.2204 -1.5123
145 97.60 7.4176 0.1824 -1.7016
150 98.00 7.4480 0.1520 -1.8839
155 98.20 7.4632 0.1368 -1.9892
160 98.50 7.4860 0.1140 -2.1716
165 98.80 7.5088 0.0912 -2.3947
170 99.00 7.5240 0.0760 -2.5770
175 99.20 7.5392 0.0608 -2.8002
180 99.30 7.5468 0.0532 -2.9337
185 99.50 7.5620 0.0380 -3.2702
190 99.60 7.5696 0.0304 -3.4933
195 99.70 7.5772 0.0228 -3.7810
200 99.70 7.5772 0.0228 -3.7810
205 99.80 7.5848 0.0152 -4.1865
210 99.80 7.5848 0.0152 -4.1865
215 99.90 7.5924 0.0076 -4.8796
220 99.90 7.5924 0.0076 -4.8796
225 99.90 7.5924 0.0076 -4.8796
230 100.00 7.6000 0.0000 -

Table 7.2.5: Table for aeration 2.5 L/min at each 5 seconds

Graph 7.2.5: Graph of ln (C*-CL) versus time for aeration 2.5 L/min
 Aeration (L/min) kLa (h-1)
0.5 36.72
1.0 45.36
1.5 73.44
2.0 95.76
2.5 115.20

Table 7.2.6: Table of kLa value for aeration at 0.5, 1.0, 1.5, 2.0 and 2.5 L/min

Graph 7.2.6: Graph of kLa versus aeration

7.3 Procedures for agitation as manipulated variable

Constant variables:
Temperature = 30°C
C* = 7.6 mg/L
Aeration = 2 L/min

Time (s) Agitation = 200 rpm

PO2% CL C*˗ CL ln (C*˗ CL)
5 2.52 0.1915 7.4085 2.0026
10 6.51 0.4948 7.1052 1.9608
15 9.01 0.6848 6.9152 1.9337
20 12.4 0.9424 6.6576 1.8958
25 15.8 1.2008 6.3992 1.8562
30 19.1 1.4516 6.1484 1.8162
35 23.1 1.7556 5.8444 1.7655
40 27.0 2.0520 5.5480 1.7134
45 30.7 2.3332 5.2668 1.6614
50 34.6 2.6296 4.9704 1.6035
55 37.5 2.8500 4.7500 1.5581
60 40.9 3.1084 4.4916 1.5022
65 44.0 3.3440 4.2560 1.4483
70 47.1 3.5796 4.0204 1.3914
75 50.6 3.8456 3.7544 1.3229
80 53.3 4.0508 3.5492 1.2667
85 56.0 4.2560 3.3440 1.2072
90 58.7 4.4612 3.1388 1.1438
95 61.2 4.6512 2.9488 1.0814
100 63.6 4.8336 2.7664 1.0175
105 65.9 5.0084 2.5916 0.9523
110 68.0 5.1680 2.4320 0.8887
115 70.0 5.3200 2.2800 0.8242
120 71.8 5.4568 2.1432 0.7623
125 73.4 5.5784 2.0216 0.7039
130 75.0 5.7000 1.9000 0.6419
135 76.5 5.8140 1.7860 0.5800
140 78.0 5.9280 1.6720 0.5140
145 79.4 6.0344 1.5656 0.4483
150 80.8 6.1408 1.4592 0.3779
155 82.0 6.2320 1.3680 0.3133
160 83.0 6.3080 1.2920 0.2562
165 84.3 6.4068 1.1932 0.1766
170 85.4 6.4904 1.1096 0.1040
175 86.1 6.5436 1.0564 0.0549
180 87.1 6.6196 0.9804 -0.0198
185 88.0 6.6880 0.9120 -0.0921
190 88.8 6.7488 0.8512 -0.1611
195 89.4 6.7944 0.8056 -0.2162
200 90.1 6.8476 0.7524 -0.2845
205 90.7 6.8932 0.7068 -0.3470
210 91.4 6.9464 0.6536 -0.4253
215 92.0 6.9920 0.6080 -0.4976
220 92.5 7.0300 0.5700 -0.5621
225 93.0 7.0680 0.5320 -0.6311
230 93.5 7.1060 0.4940 -0.7052
235 94.0 7.1440 0.4560 -0.7853
240 94.4 7.1744 0.4256 -0.8543
245 94.8 7.2048 0.3952 -0.9284
250 95.1 7.2276 0.3724 -0.9878
255 95.4 7.2504 0.3496 -1.0510
260 95.8 7.2808 0.3192 -1.1419
265 96.1 7.3036 0.2964 -1.2160
270 96.4 7.3264 0.2736 -1.2961
275 96.7 7.3492 0.2508 -1.3831
280 96.9 7.3644 0.2356 -1.4456
285 97.1 7.3796 0.2204 -1.5123
290 97.3 7.3948 0.2052 -1.5838
295 97.5 7.4100 0.1900 -1.6607
300 97.7 7.4252 0.1748 -1.7441
305 97.9 7.4404 0.1596 -1.8351
310 98.0 7.4480 0.1520 -1.8839
315 98.1 7.4556 0.1444 -1.9352
320 98.2 7.4632 0.1368 -1.9892
325 98.3 7.4708 0.1292 -2.0464
330 98.5 7.4860 0.1140 -2.1716
335 98.6 7.4936 0.1064 -2.2405
340 98.8 7.5088 0.0912 -2.3947
345 98.9 7.5164 0.0836 -2.4817
350 99.1 7.5316 0.0684 -2.6824
355 99.2 7.5392 0.0608 -2.8002
360 99.3 7.5468 0.0532 -2.9337
365 99.3 7.5468 0.0532 -2.9337
370 99.3 7.5468 0.0532 -2.9337
375 99.4 7.5544 0.0456 -3.0878
380 99.4 7.5544 0.0456 -3.0878
385 99.5 7.5620 0.0380 -3.2702
390 99.6 7.5696 0.0304 -3.4933
395 99.6 7.5696 0.0304 -3.4933
400 99.7 7.5772 0.0228 -3.7810
405 99.7 7.5772 0.0228 -3.7810
410 99.7 7.5772 0.0228 -3.7810
415 99.7 7.5772 0.0228 -3.7810
420 99.7 7.5772 0.0228 -3.7810

Table 7.3.1: Table for agitation 200 rpm at each 5 seconds

Graph 7.3.1: Graph of ln (C*-CL) versus time for agitation 200 rpm
Time (s) Agitation = 400 rpm
PO2% CL C*˗ CL ln (C*˗ CL)
5 2.59 0.1968 7.4032 2.0019
10 6.69 0.5084 7.0916 1.9589
15 12.6 0.9576 6.6424 1.8935
20 19.0 1.4440 6.1560 1.8174
25 25.5 1.9380 5.6620 1.7338
30 32.3 2.4548 5.1452 1.6381
35 39.9 3.0324 4.5676 1.5190
40 45.1 3.4276 4.1724 1.4285
45 50.7 3.8532 3.7468 1.3209
50 57.4 4.3624 3.2376 1.1748
55 61.0 4.6360 2.9640 1.0865
60 65.3 4.9628 2.6372 0.9697
65 69.0 5.2440 2.3560 0.8570
70 73.0 5.5480 2.0520 0.7188
75 76.1 5.7836 1.8164 0.5969
80 78.0 5.9280 1.6720 0.5140
85 81.6 6.2016 1.3984 0.3353
90 83.6 6.3536 1.2464 0.2203
95 85.4 6.4904 1.1096 0.1040
100 87.1 6.6196 0.9804 -0.0198
105 88.8 6.7488 0.8512 -0.1611
110 90.1 6.8476 0.7524 -0.2845
115 91.5 6.9540 0.6460 -0.4370
120 92.6 7.0376 0.5624 -0.5755
125 93.4 7.0984 0.5016 -0.6900
130 94.3 7.1668 0.4332 -0.8366
135 95.0 7.2200 0.3800 -0.9676
140 95.6 7.2656 0.3344 -1.0954
145 96.2 7.3112 0.2888 -1.2420
150 96.6 7.3416 0.2584 -1.3532
155 97.4 7.4024 0.1976 -1.6215
160 97.8 7.4328 0.1672 -1.7886
165 98.0 7.4480 0.1520 -1.8839
170 98.2 7.4632 0.1368 -1.9892
175 98.3 7.4708 0.1292 -2.0464
180 98.5 7.4860 0.1140 -2.1716
185 98.7 7.5012 0.0988 -2.3147
190 99.0 7.5240 0.0760 -2.5770
195 99.1 7.5316 0.0684 -2.6824
200 99.2 7.5392 0.0608 -2.8002
205 99.3 7.5468 0.0532 -2.9337
210 99.4 7.5544 0.0456 -3.0878
215 99.5 7.5620 0.0380 -3.2702
220 99.6 7.5696 0.0304 -3.4933
225 99.7 7.5772 0.0228 -3.7810
230 99.8 7.5848 0.0152 -4.1865
235 99.9 7.5924 0.0076 -4.8796
240 99.9 7.5924 0.0076 -4.8796
245 100 7.6000 0.0000 -

Table 7.3.2: Table for agitation 400 rpm at each 5 seconds

Graph 7.3.2: Graph of ln (C*-CL) versus time for agitation 400 rpm
Time (s) Agitation = 600 rpm
PO2% CL C*˗ CL ln (C*˗ CL)
5 4.8 0.3648 7.2352 1.9790
10 15.6 1.1856 6.4144 1.8585
15 45.0 3.4200 4.1800 1.4303
20 50.5 3.8380 3.7620 1.3250
25 58.1 4.4156 3.1844 1.1583
30 65.1 4.9476 2.6524 0.9755
35 70.3 5.3428 2.2572 0.8141
40 76.1 5.7836 1.8164 0.5969
45 80.3 6.1028 1.4972 0.4036
50 83.9 6.3764 1.2236 0.2018
55 86.5 6.5740 1.0260 0.0257
60 88.9 6.7564 0.8436 -0.1701
65 91.2 6.9312 0.6688 -0.4023
70 92.9 7.0604 0.5396 -0.6169
75 93.9 7.1364 0.4636 -0.7687
80 95.1 7.2276 0.3724 -0.9878
85 96.0 7.2960 0.3040 -1.1907
90 96.7 7.3492 0.2508 -1.3831
95 97.3 7.3948 0.2052 -1.5838
100 97.8 7.4328 0.1672 -1.7886
105 98.2 7.4632 0.1368 -1.9892
110 98.5 7.4860 0.1140 -2.1716
115 98.9 7.5164 0.0836 -2.4817
120 99.1 7.5316 0.0684 -2.6824
125 99.3 7.5468 0.0532 -2.9337
130 99.4 7.5544 0.0456 -3.0878
135 99.6 7.5696 0.0304 -3.4933
140 99.7 7.5772 0.0228 -3.7810
145 99.8 7.5848 0.0152 -4.1865
150 99.9 7.5924 0.0076 -4.8796
 155 100 7.6000 0.0000 -

Table 7.3.3: Table for agitation 600 rpm at each 5 seconds

Graph 7.3.3: Graph of ln (C*-CL) versus time for agitation 600 rpm

Time (s) Agitation = 800 rpm

PO2% CL C*˗ CL ln (C*˗ CL)
5 9.5 0.7220 6.8780 1.9283
10 25.7 1.9532 5.6468 1.7311
15 37.2 2.8272 4.7728 1.5629
20 49.4 3.7544 3.8456 1.3469
25 61.2 4.6512 2.9488 1.0814
30 69.8 5.3048 2.2952 0.8308
35 77.7 5.9052 1.6948 0.5276
40 82.4 6.2624 1.3376 0.2909
45 86.4 6.5664 1.0336 0.0330
50 90.1 6.8476 0.7524 -0.2845
55 92.1 6.9996 0.6004 -0.5102
60 93.6 7.1136 0.4864 -0.7207
65 95.1 7.2276 0.3724 -0.9878
70 96.2 7.3112 0.2888 -1.2420
75 97.1 7.3796 0.2204 -1.5123
80 97.7 7.4252 0.1748 -1.7441
85 98.1 7.4556 0.1444 -1.9352
90 98.5 7.4860 0.1140 -2.1716
95 98.8 7.5088 0.0912 -2.3947
100 99.1 7.5316 0.0684 -2.6824
105 99.3 7.5468 0.0532 -2.9337
110 99.5 7.5620 0.0380 -3.2702
115 99.6 7.5696 0.0304 -3.4933
120 99.7 7.5772 0.0228 -3.7810
125 99.8 7.5848 0.0152 -4.1865
130 99.9 7.5924 0.0076 -4.8796
135 100 7.6000 0.0000 -

Table 7.3.4: Table for agitation 800 rpm at each 5 seconds

Graph 7.3.4: Graph of ln (C*-CL) versus time for agitation 800 rpm
 Agitation = 1000 rpm
Time (s) PO2% CL C*˗ CL ln (C*˗ CL)
5 9.1 0.6916 6.9084 1.9327
10 25.0 1.9000 5.7000 1.7405
15 45.8 3.4808 4.1192 1.4157
20 57.2 4.3472 3.2528 1.1795
25 67.8 5.1528 2.4472 0.8949
30 76.7 5.8292 1.7708 0.5714
35 82.7 6.2852 1.3148 0.2737
40 87.5 6.6500 0.9500 -0.0513
45 90.5 6.8780 0.7220 -0.3257
50 93.2 7.0832 0.5168 -0.6601
55 94.8 7.2048 0.3952 -0.9284
60 95.2 7.2352 0.3648 -1.0084
65 97.0 7.3720 0.2280 -1.4784
70 97.8 7.4328 0.1672 -1.7886
75 98.2 7.4632 0.1368 -1.9892
80 98.7 7.5012 0.0988 -2.3147
85 99.1 7.5316 0.0684 -2.6824
90 99.3 7.5468 0.0532 -2.9337
95 99.6 7.5696 0.0304 -3.4933
100 99.7 7.5772 0.0228 -3.7810
105 99.9 7.5924 0.0076 -4.8796
110 100 7.6000 0.0000 -

Table 7.3.5: Table for agitation 1000 rpm at each 5 seconds

 Graph 7.3.5: Graph of ln (C*-CL) versus time for agitation 1000 rpm

Agitation (rpm) kLa (h-1)

200 51.48
400 99.36
600 154.08
800 185.04
1000 223.20

Table 7.3.6: Table of kLa value for agitation at 200, 400, 600, 800 and 1000 rpm

Graph 7.3.6: Graph of kLa versus agitation

8.0 CALCULATIONS

A plot of ln (C*˗ CL) versus time, t should result in a straight line of slope –kLa (Moutafchieva,
Popova, Dimitrova, & Tchaoushev, 2013, 48(4)).

ln(𝐶 ∗ − 𝐶𝐿 ) = − 𝑘𝐿 𝑎𝑡 (Equation 8.1)

𝑦 = 𝑚𝑥 + 𝐶 (Equation 8.2)

Compare equation 8.1 and equation 8.2,

∴ 𝑦 = ln(𝐶 ∗ − 𝐶𝐿 ) , 𝑥 = 𝑡, 𝑚 (𝑠𝑙𝑜𝑝𝑒) = −𝑘𝐿 𝑎

Sample of calculations:

For temperature 35°C, C* = 7 mg/L,

At t = 5 s,

PO2% = 0.20%

𝐷𝑂 𝑚𝑔
𝐶𝐿 = ×7
100 𝐿

0.20 𝑚𝑔
𝐶𝐿 = ×7
100 𝐿
𝑚𝑔
𝐶𝐿 (𝐷𝑂) = 0.0140
𝐿

𝑚𝑔 𝑚𝑔
𝐶 ∗ − 𝐶𝐿 = 7 − 0.014 = 6.9860 𝑚𝑔/𝐿
𝐿 𝐿

ln(𝐶 ∗ − 𝐶𝐿 ) = ln(6.9860)

ln(𝐶 ∗ − 𝐶𝐿 ) = 1.9439
For temperature at 35°C:

Based on the graph 7.1.1 plotted,

𝑠𝑙𝑜𝑝𝑒 = −0.0413 𝑠 −1

−𝑘𝐿 𝑎 = −0.0413 𝑠 −1

0.0413 60 𝑚𝑖𝑛 60 𝑠
𝑘𝐿 𝑎 = × ×
1𝑠 1ℎ 1 𝑚𝑖𝑛

𝒌𝑳 𝒂 = 𝟏𝟒𝟖. 𝟔𝟖 𝒉−𝟏

For temperature at 40°C:

Based on the graph 7.1.2 plotted,

𝑠𝑙𝑜𝑝𝑒 = −0.0507 𝑠 −1

−𝑘𝐿 𝑎 = −0.0507 𝑠 −1

0.0507 60 𝑚𝑖𝑛 60 𝑠
𝑘𝐿 𝑎 = × ×
1𝑠 1ℎ 1 𝑚𝑖𝑛

𝒌𝑳 𝒂 = 𝟏𝟖𝟐. 𝟓𝟐 𝒉−𝟏

For temperature at 45°C:

Based on the graph 7.1.3 plotted,

𝑠𝑙𝑜𝑝𝑒 = −0.0609 𝑠 −1

−𝑘𝐿 𝑎 = −0.0609 𝑠 −1

0.0609 60 𝑚𝑖𝑛 60 𝑠
𝑘𝐿 𝑎 = × ×
1𝑠 1ℎ 1 𝑚𝑖𝑛

𝒌𝑳 𝒂 = 𝟐𝟏𝟗. 𝟐𝟒 𝒉−𝟏

For temperature at 50°C:

Based on the graph 7.1.4 plotted,

𝑠𝑙𝑜𝑝𝑒 = −0.0657 𝑠 −1

−𝑘𝐿 𝑎 = −0.0657 𝑠 −1
 0.0657 60 𝑚𝑖𝑛 60 𝑠
𝑘𝐿 𝑎 = × ×
1𝑠 1ℎ 1 𝑚𝑖𝑛

𝒌𝑳 𝒂 = 𝟐𝟑𝟔. 𝟓𝟐 𝒉−𝟏

For aeration at 0.5 L/min:

Based on the graph 7.2.1 plotted,

𝑠𝑙𝑜𝑝𝑒 = −0.0102 𝑠 −1

−𝑘𝐿 𝑎 = −0.0102 𝑠 −1

0.0102 60 𝑚𝑖𝑛 60 𝑠
𝑘𝐿 𝑎 = × ×
1𝑠 1ℎ 1 𝑚𝑖𝑛

𝒌𝑳 𝒂 = 𝟑𝟔. 𝟕𝟐 𝒉−𝟏

For aeration at 1.0 L/min:

Based on the graph 7.2.2 plotted,

𝑠𝑙𝑜𝑝𝑒 = −0.0126 𝑠 −1

−𝑘𝐿 𝑎 = −0.0126 𝑠 −1

0.0126 60 𝑚𝑖𝑛 60 𝑠
𝑘𝐿 𝑎 = × ×
1𝑠 1ℎ 1 𝑚𝑖𝑛

𝒌𝑳 𝒂 = 𝟒𝟓. 𝟑𝟔 𝒉−𝟏

For aeration at 1.5 L/min:

Based on the graph 7.2.3 plotted,

𝑠𝑙𝑜𝑝𝑒 = −0.0204 𝑠 −1

−𝑘𝐿 𝑎 = −0.0204 𝑠 −1

0.0204 60 𝑚𝑖𝑛 60 𝑠
𝑘𝐿 𝑎 = × ×
1𝑠 1ℎ 1 𝑚𝑖𝑛
 𝒌𝑳 𝒂 = 𝟕𝟑. 𝟒𝟒 𝒉−𝟏

For aeration at 2.0 L/min:

Based on the graph 7.2.4 plotted,

𝑠𝑙𝑜𝑝𝑒 = −0.0266 𝑠 −1

−𝑘𝐿 𝑎 = −0.0266 𝑠 −1

0.0266 60 𝑚𝑖𝑛 60 𝑠
𝑘𝐿 𝑎 = × ×
1𝑠 1ℎ 1 𝑚𝑖𝑛

𝒌𝑳 𝒂 = 𝟗𝟓. 𝟕𝟔 𝒉−𝟏

For aeration at 2.5 L/min:

Based on the graph 7.2.5 plotted,

𝑠𝑙𝑜𝑝𝑒 = −0.0320 𝑠 −1

−𝑘𝐿 𝑎 = −0.0320 𝑠 −1

0.0320 60 𝑚𝑖𝑛 60 𝑠
𝑘𝐿 𝑎 = × ×
1𝑠 1ℎ 1 𝑚𝑖𝑛

𝒌𝑳 𝒂 = 𝟏𝟏𝟓. 𝟐𝟎 𝒉−𝟏

For agitation at 200 rpm:

Based on the graph 7.3.1 plotted,

𝑠𝑙𝑜𝑝𝑒 = −0.0143 𝑠 −1

−𝑘𝐿 𝑎 = −0.0143 𝑠 −1

0.0143 60 𝑚𝑖𝑛 60 𝑠
𝑘𝐿 𝑎 = × ×
1𝑠 1ℎ 1 𝑚𝑖𝑛

𝒌𝑳 𝒂 = 𝟓𝟏. 𝟒𝟖 𝒉−𝟏

For agitation at 400 rpm:

Based on the graph 7.3.2 plotted,

𝑠𝑙𝑜𝑝𝑒 = −0.0276 𝑠 −1
 −𝑘𝐿 𝑎 = −0.0276 𝑠 −1

0.0276 60 𝑚𝑖𝑛 60 𝑠
𝑘𝐿 𝑎 = × ×
1𝑠 1ℎ 1 𝑚𝑖𝑛

𝒌𝑳 𝒂 = 𝟗𝟗. 𝟑𝟔 𝒉−𝟏

For agitation at 600 rpm:

Based on the graph 7.3.3 plotted,

𝑠𝑙𝑜𝑝𝑒 = −0.0428 𝑠 −1

−𝑘𝐿 𝑎 = −0.0428 𝑠 −1

0.0428 60 𝑚𝑖𝑛 60 𝑠
𝑘𝐿 𝑎 = × ×
1𝑠 1ℎ 1 𝑚𝑖𝑛

𝒌𝑳 𝒂 = 𝟏𝟓𝟒. 𝟎𝟖 𝒉−𝟏

For agitation at 800 rpm:

Based on the graph 7.3.4 plotted,

𝑠𝑙𝑜𝑝𝑒 = −0.0514 𝑠 −1

−𝑘𝐿 𝑎 = −0.0514 𝑠 −1

0.0514 60 𝑚𝑖𝑛 60 𝑠
𝑘𝐿 𝑎 = × ×
1𝑠 1ℎ 1 𝑚𝑖𝑛

𝒌𝑳 𝒂 = 𝟏𝟖𝟓. 𝟎𝟒 𝒉−𝟏

For agitation at 1000 rpm:

Based on the graph 7.3.5 plotted,

𝑠𝑙𝑜𝑝𝑒 = −0.0620 𝑠 −1

−𝑘𝐿 𝑎 = −0.0620 𝑠 −1

0.0620 60 𝑚𝑖𝑛 60 𝑠
𝑘𝐿 𝑎 = × ×
1𝑠 1ℎ 1 𝑚𝑖𝑛

𝒌𝑳 𝒂 = 𝟐𝟐𝟑. 𝟐𝟎 𝒉−𝟏
9.0 DISCUSSION

The aims of this experiment are to measure the volumetric mass transfer coefficient,
kLa of stirred tank reactor with bubble aeration and to study the relationship between
temperature, aeration and agitation between the kLa values in bioreactor. Due to the absence of
cells, for this experiment, the method used to measure the kLa value is by using the unsteady-
state method, which is shown in equations below.

𝑑𝐶𝐿
= 𝑂𝑇𝑅 − 𝑂𝑈𝑅
𝑑𝑡

Since OUR = 0, the equation is simplified as below.

𝑑𝐶𝐿
= 𝑂𝑇𝑅 = 𝑘𝐿 𝑎(𝐶 ∗ − 𝐶𝐿 )
𝑑𝑡

−𝑑(𝐶 ∗ − 𝐶𝐿 )
= 𝑘𝐿 𝑎
𝑑𝑡/(𝐶 ∗ − 𝐶𝐿 )

ln(𝐶 ∗ − 𝐶𝐿 ) = − 𝑘𝐿 𝑎𝑡

The equation obtained is similar with the linear equation y = mx + C. Hence, the graph of ln
(C*-CL) versus time, t will obtain a straight line with a slope of –kLa. Before the experiment
was conducted, the bioreactor’s probe was done with 2 points calibration to prevent errors that
can affect the results.

To study the effect of temperature towards the k La value, different temperatures were
used to conduct this experiment, which are 35°C, 40°C, 45°C and 50°C. On the other hand, the
bioreactor’s parameter such as aeration and agitation were set fixed which were at 2 L/min and
400 rpm respectively for each reading. The concentration of dissolved oxygen was recorded
for each 5 seconds until the concentration of dissolved oxygen in the bioreactor is saturated.
The graph of ln (C*-CL) versus time, was plotted to determine the kLa value. For the
temperature at 35°C, 40°C, 45°C and 50°C, the kLa value obtained are 148.68 h-1, 182.52 h-1,
219.24 h-1 and 236.52 h-1. From the graph 7.1.5, it shows that the higher temperature, the higher
the mass transfer coefficient. Besides, the time taken for the concentration of dissolve oxygen
to become saturated is shorter when high temperature is used. The concentration of the
undissolved oxygen for each temperature decreases as the time goes. However, the
concentration of the dissolve oxygen obtained was over 100%, this is due to several errors done
during the calibration.
 To study the effect of aeration rate towards the kLa value, different values of aeration
were used to conduct this experiment. The aeration rate used was started with 0.5 L/min and
continued with 1.0, 1.5, 2.0 and 2.5 L/min. The bioreactor’s parameter such as temperature and
agitation are set fixed at 30°C and 400 rpm respectively for each reading. The value of dissolve
oxygen was recorded for every 5 seconds. To determine the kLa value, the graph of ln (C*-CL)
versus time was plotted. For the aeration rate at 0.5, 1.0, 1.5, 2.0 and 2.5 L/min, the kLa obtained
is 36.72 h-1, 45.36 h-1, 73.44 h-1, 95.76 h-1 and 115.20 h-1 respectively. From the graph 7.2.6
plotted, it shows that the higher the aeration rate, the higher the k La value. The time taken for
the concentration of dissolve oxygen to become saturated is also shorter as the rate of aeration
increases. This is because when the oxygen transfer rate is high, the movement of particles will
be faster and more efficient, so the faster the oxygen dissolve and become saturated. The mass
transfer coefficient is higher when higher aeration rate is used.

To study the effect of agitation speed towards the kLa value, five different values of
agitation rate were used, which are 200, 400, 600, 800 and 1000 rpm. The graph of ln (C*-CL)
versus time, was plotted to determine the kLa value. For the agitation speed at 200 rpm, 400
rpm, 600 rpm, 800 rpm and 1000 rpm, the kLa value obtained is 51.48 h-1, 99.36 h-1, 154.08 h-
1
, 185.04 h-1 and 223.2 h-1 respectively. From the graph 7.3.6 plotted, it shows that the higher
the agitation speed, the higher the mass transfer coefficient. For low agitation rates, which is
200 rpm, the turbulence is not enough to trap and hold up the air bubbles and consequently the
performance by the volumetric mass transfer rate may not increase noticeably. Therefore,
agitation rate of 400 rpm and above is suitable for the future bioprocess operations.

The manipulated variable that affects the mass transfer coefficient the most is the
temperature. However, low aeration rate and low agitation speed are not favoured in bioprocess
operations. For every manipulated variable, as the value gets higher, the time taken for the
concentration of dissolve oxygen to become saturated is shorter.

There are several errors that had been made during the experiment. Firstly, it might
cause by the conditions of the bioreactor. Although the value of 100 was set during the
calibration, the reading of the dissolve oxygen still exceeded the set point of calibration.
Therefore, this will affect the value of C* at the specific temperature and caused the result to
be less accurate. Next, there might be some impurities in the bioreactor that can affect the
results. Moreover, the transfer of oxygen from gas phase to liquid phase is complicated by
presence of cells, product formation, ionic species and anti-foaming agents. Furthermore, the
ambient temperature and the disturbances from the surrounding could be effect of the errors.
The amount of oxygen might not fully dissolved in the reaction.

There are a few ways to overcome the errors. Firstly, the bioreactor should be made
sure that it is in well condition before the experiment. Besides, the bioreactor should be cleaned
properly before it is used for the experiment because the impurities may affect the readings.
Next, the experiment should be done in a surrounding with less disturbances. Furthermore, the
calibration should be done correctly to avoid errors from happening.

There are a few precautions that should be alert in this experiment. Firstly, safety
goggles should be worn throughout the experiment to avoid any gas or explosion to get contact
with the eyes. Next, avoid pointing the nitrogen gas pipe towards the eyes because high
pressure of purging nitrogen gas into the reactor can result the nitrogen gas to burst from the
pipe. The nitrogen gas tank valve should be adjusted carefully to avoid the nitrogen gas bursting
out from the pipe to the surrounding.
10.0 CONCLUSION

In conclusion, the unsteady-state method was used to measure the value of volumetric
mass transfer coefficient, kLa of a stirred tank reactor with bubble aeration. The value of kLa
are dependent with temperature, aeration and agitation. 2 points calibration was done before
purging the nitrogen gas into the system. The reading of dissolve oxygen was taken in every 5
seconds until it becomes saturated. The graph of ln (C*-CL) versus time, was plotted to
determine the kLa value and the graph of temperature, aeration and agitation versus kLa were
also plotted to observe their relationship. To study the effect of temperature towards the k La
value, different temperatures were used to conduct the experiment. For the temperature at 35°C,
40°C, 45°C and 50°C, the kLa value obtained are 148.68 h-1, 182.52 h-1, 219.24 h-1 and 236.52
h-1. From the graph 7.1.5, it shows that the higher temperature, the higher the mass transfer
coefficient. To study the effect of aeration rate towards the k La value, different values of
aeration were used to conduct this experiment. For the aeration rate at 0.5, 1.0, 1.5, 2.0 and 2.5
L/min, the kLa obtained is 36.72 h-1, 45.36 h-1, 73.44 h-1, 95.76 h-1 and 115.20 h-1 respectively.
From the graph 7.2.6 plotted, it shows that the higher the aeration rate, the higher the kLa value.
To study the effect of agitation speed towards the kLa value, five different values of agitation
rate were used, which are 200, 400, 600, 800 and 1000 rpm. For the agitation speed at 200 rpm,
400 rpm, 600 rpm, 800 rpm and 1000 rpm, the kLa value obtained is 51.48 h-1, 99.36 h-1, 154.08
h-1, 185.04 h-1 and 223.2 h-1 respectively. From the graph 7.3.6 plotted, it shows that the higher
the agitation speed, the higher the mass transfer coefficient. Besides, it was observed that the
increase in temperature, aeration rate and agitation speed resulted with shorter time for the
dissolve oxygen to reach the saturated level. Hence, the aims are successfully achieved and the
experiment was conducted successfully.
11.0 RECOMMENDATIONS

There are a few of recommendations that could improve the result of this experiment.
Firstly, the experiment should be repeated for at least 3 times to obtain more accurate value of
the mass transfer coefficient. Besides, before conducting the experiment, the bioreactor’s probe
must be made sure that it has been calibrated to prevent errors that could affect the results of
the experiment. Next, the data collected has to be made sure that it is collected at every 5
seconds to ensure the correct data is taken as the value of dissolve oxygen changes. To achieve
a more accurate result, the readings should be taken until five stable values of dissolve oxygen
is achieved.
12.0 REFERENCE
Moutafchieva, D., Popova, D., Dimitrova, M., & Tchaoushev, S. (2013, 48(4)). Experimental
Determination of the Volumetric Mass Transfer Coefficient. Journal of Chemical
Technology and Metallurgy, 351-356.

Pinelli, D., Liu, Z., & Magelli, F. (2010, 8). Analysis of KLa Measurement Methods in Stirred
Vessels: The Role of Experimental Techniques and Fluid Dynamic Models.
International Journal of Chemical Reactor Engineering, 1-38.

Sathish, S., & Amaranth, D. J. (2015, 7(3)). Experimental studies on determination of

volumetric oxygen transfer coefficient in stirred tank bioreactor using fungal broths
isolated from textile effluent. Journal of Chemical and Pharmaceutical Research, 814-
818.

Shuler, M. L., & Kargi, F. (2002). Bioprocess Engineering Basic Concepts (Second Edition).
New Jersey: Prentice Hall PTR.

Zedníková, M., Orvalho, S., Fialová, M., & Ruzicka, M. C. (2018, 2(19)). Measurement of
Volumetric Mass Transfer Coefficient in Bubble Columns. ChemEngineering, 1-14.
13.0 APPENDIX

Figure 13.1: Linear air pump aerator

Figure 13.2: Pipe mouth Figure 13.3: Nitrogen gas tank

where the nitrogen gas is
purged