European

J..... ,o,Applied
9

European J. Appl. Microbiol.

Biotechnol. 6, 3 1 5 - 3 2 3 (1979) Microbiology a n d
Biotechnology
9 by S p r i n g e r - V e r l a g 9 1979

Methods for Measuring the Bubble Size

in Bubble Column Bioreactors II

R. Buchholz and K. Schiigerl

Institut far Technische Chemie der Universitiit Hannover, Callinstrage 3, D-3000 Hannover 1,
Federal Republic of Germany

Summary. Electrooptical and light scattering methods for measuring the

structure of two phase flow, especially bubble size, are described and compared
with photographic and electrical conductivity methods.

1. Introduction

In the foregoing paper (part I) (Buchholz and Schiigerl, 1979) photographic and elec-
trical conductivity methods were discussed. In the present paper two further methods
are considered, the electrooptical and light scattering methods. The latter was not in-
vestigated by the authors. However, based on the literature data, it will also be dis-
cussed and compared with other methods.

2. Electrooptical Method

The electrooptical probe was developed by Todtenhaupt (1971) and Pilhofer (Pilhofer
and Miller, 1972) (Pilhofer, 1974) as well as by Jekat (1975) and improved by Brentrup
et al. (1977). Figure 1 shows the principle of this method. A negatively pressurized
capillary tube, which is equipped with a funnel inlet draws off the two phase system.
In the capillary tube, the bubble is deformed into a cylindrical plug which passes two
light gates. By passing the gas/liquid interface through the gate, the intensity of the
light at the detector is at first increased, then decreased and increased again due to
focusing deflection and focusing of the light beam. At gates 1 and 2 the plug causes
near square-wave signals with the durations At 1 and ~t 2. The time lag between the two
signals is ZXtl2. The bubble volume is given, neglecting the curvatures of the plug at its
two ends, by Eq. (1):

V B = h A D ZXt1/At12 (1)

where h is a calibration constant and A D the cross section area of the capillary. Since
Eq. (1), used for the evaluation of the measurements made by the electrooptical

0171--1741/79/0006/0315/~ 01.80
316 R. Buchholz and K. Schiigerl

f,-,
Ate2

T
(b)
(a)
~;Z. b

Fig. 1. a. Electrooptical sensor (Pilhofer and Miiller, 1972). a = capillary; b = inlet funnel; c = light
source; d = light detector; e = bubble; f = liquid, distance of the gates 1 and 2; b = 132 mm.
course of light beam with liquid in the capillary; - - - course of light beam with bubble in
the capillary, b. Signals at the gates 1 and 2

method, and Eq. (2b) of part I (Buchholz and Schiigerl, 1979), used for the evalua-
tion of the measurements carried out b y the electrical conductivity method, are
analogue, the same electronics (Fig. 3 in part I) are applied for both of these probes.
The application of this probe can be limited b y the following shortcomings: at high
removal rates the bubbles are dispersed in the inlet funnel and/or in the capillary tube;
at low removal rates the bubbles can be dispersed if t h e y are much larger than the inlet
funnel; at too low removal rates and in a coalescence promoting media, the bubbles
can coalesce in the inlet funnel; b y using a capillary with diameter larger than the
bubble diameter, no signal processing is possible; only by an isokinetic removal rate
is segregation of the bubbles with different diameters possible. However, isokinetic
removal rate promotes coalescence; at too high bubble concentration a separation of
the signals caused by different bubbles is not possible.
For bubbles with narrow size distributions, most of these difficulties can be
avoided b y using a suitable capillary diameter and removal rate. In case of bubbles with
broad size distributions, no satisfactory solution is possible, if one uses only a single
capillary diameter: in a capillary with a medium diameter, the small bubbles cannot
be detected and the large bubbles are dispersed. To evaluate the size distribution of
bubbles with broad size distribution, the use of capillaries with different diameters
is necessary.
By using focusing lenses to improve resolution of the light beam (Brentrup, 1977),
b y applying capillary tubes with an inner surface which was made hydrophobic to
diminish the hazard to bubble dispersion, by reducing the shear stress in the capillary,
and, to improve the signal quality, by eliminating the liquid film which adheres to the
capillary surface causing noisy base lines (Buchholz, 1978), the quality of this meas-
uring technique was significantly improved. The hydrophobic surface made it possible
to use high removal rates without dispersing the bubbles in the capillary.
Bubble C o l u m n Bioreactors II 317

The measurements of the present paper were m a d e b y an electrooptical probe, which

was developed b y Brentrup and O n k e n (1977) and placed b y U. O n k e n at our disposal.
The same electronics were used for these measurements which were developed for the
double electrical c o n d u c t i v i t y probe (Buchholz, 1979). Since in most of the investigated
systems the bubbles had narrow size distribution, a single suitable capillary diameter
for a given system was used for t h e measurements, e.g., 1.5 m m for distilled water and
0.5 and 1.13 m m for alcohol solutions at different distances f r o m t h e aerator.

3. Light Scattering Method

Light scattering m e t h o d s were used a long time ago. Sauter (1928) first d e t e r m i n e d
the interfacial areas of carburettor sprays by the use of a light-obscuration m e t h o d .
Vermeulen (1955) and Trice et al. (1956) used light transmission to d e t e r m i n e the in-
teffacial areas of gas/liquid and liquid/liquid dispersions. Calderbank (1958) adapted
Sauter's t e c h n i q u e and later (Calderbank, 1960) described a m e t h o d for measuring the
interfacial area of dense dispersions f r o m their optical reflectivity. A review of these
m e t h o d s is given b y Calderbank (1967). All of these m e t h o d s yield integral values.
To evaluate t h e local structure o f t h e t w o phase f l o w of organic liquids, t h e use of fi-
ber optics is advantageous.
Geake et al. (1975) used optical sensors (dip-sticks) to sense changes in the refrac-
tive index at their e x t r e m i t y . Calderbank and Pereira (1977) c o m b i n e d five miniature
versions of these optical "dip-sticks" into a sensor similar to the electrical c o n d u c t i v i t y
sensors consisting of five electrodes developed b y Burgess and Calderbank (1975). In
this c o m p o u n d p r o b e (Fig. 2), the distance b e t w e e n the central sensor 1 and the sensor
2, 3 or 4 is 0.103 cm and the distance b e t w e e n the sensors i and 5 is 0.495 cm. Hence,
o n l y bubbles larger than 1 c m ' i n d i a m e t e r can be detected. If sensor 5 is disregarded,
bubbles 0.2 cm in m i n i m u m diameter can also be registered. A similar probe, but with
o n l y two sensors, is available on the market (DISA T y p e 55 S optical sensor, for bub-
bles 1 m m m i n i m u m diameter).

120~
Fig. 2. Combination of five optical sensors (dip-sticks)
(Calderbank and Pereira, 1977). k = fiber optics; l = flange; m =
spacer; n = stainless steel hypodermic tubing; o = electrode tip;
p = stainless steel tube ; q = glass electrode. Longitudinal distance
from the tip of sensor i to the tip of sensor 2, 3 or 4: a = 2.93 mm;
transverse distance from the tip of sensor 1 to the tip of sensor 2,
3 or 4: b = 1.03 mm; transverse distance from the tip of sensor 1
to the tip of sensor 5: c = 4.95 mm; length of the sensor tip: d =
1 4 . 2 1 m m ; e = 8 c m , f = 2 4 c m , g = 5 cm, b=O.079cm, i=
b; c-
0.398 cm (i
318 R. Buchholz and K. Schiigerl

tOO

0.80

060

040

0.20

0,00
I I
1.00
I
JY
200 3.00 &O0 5.00 6,00
• p
700
d(mm]

Fig. 3. Distribution of bubble diameter estimated by the electrooptical sensor. Bubble column 14
cm in diameter, 267 cm high. Porous plate distribution, 50 #m mean pore diameter. Distilled
water. Superficial gas velocity 1.07 cm s-1. Distance from the gas distributor 132 cm. d = 3.2 cm,
d s = 3.45 cm

4. Practical Advice for t h e Use o f Electrooptical Sensors

Figure 3 shows a b u b b l e size d i s t r i b u t i o n which was measured in distilled water b y

an electrooptical sensor with a sensor capillary diameter of 1.5 ram.

Critical Points: The light b e a m across t h e capillary must be exactly adjusted; the
Schmitt trigger threshold voltage must be set for b o t h channels to 40% of m a x i m u m
voltage; the sensor capillary diameter must be selected carefully according to the bub-
ble size distribution. F o r narrow b u b b l e size distributions, a single capillary is adequate.
The capillary diameter should be a b o u t half of the m e a n b u b b l e diameter. The use of
too small a sensor capillary diameter causes b u b b l e redistribution in the inlet funnel.
By t h e application of too large a capillary diameter, signals caused b y b u b b l e s with
diameters smaller t h a n the sensor capillary c a n n o t be separated; for wide b u b b l e size
distributions, two or more sensor capillaries m u s t be used. The construction o f the
overall b u b b l e size distribution b y means of particular ones, evaluated b y the differ-
ent sensor capillaries, is fairly difficult; the inside surface of the inlet f u n n e l and the
sensor capillary should be made hydrophobic. This avoids dispersion of the bubbles
inside the capillary at high sucking velocities due to the adhesion of the liquid o n the
capillary surface, yielding high shear velocities at the wall.

5. Comparison of the Methods

5.1 Electrooptical Method

Sensor calibrations were carried out iN the same rectangulzr euvette that was used to
calibrate electrical conductivity sensors. Single bubbles of near u n i f o r m sizes were
Bubble Column Bioreactors II 319

n_~
}-n/Ad
~,~

o,6

r -

iii/
o.2

I 2 3 4 5 6 7 8
wSG=2fl3 cm/s d(mm)

n___
o,a

(2,6 /"

.iI
/I
0,~ ,./ /
/ /

0 1 2 3 4 5 6 7
w = 1,~Tcralz d(rora,~

Fig. 4. Comparison of the bubble size distributions measured at the wall by photographic, electri-
cal conductivity - and electrooptical methods. Distilled water, h = 132 cm, WSL = 1.8 cm/s, I. At
low superficial gas velocities, wSG. - - electrooptical method; - - - photographic method;
. . . . electrical conductivity method

produced using a capillary (aerator) of definite diameter. The bubble sizes were esti-
mated by measuring the number and total volume of many hundred bubbles. The bub-
ble size was altered by varying the capillary (aerator) diameter. The measurements
showed that for the 1.5 mm (inside diameter) sensor capillary, which was used for
electrooptical measurements in distilled water, Eq. (1) did not hold for bubbles less
than 2.5 mm in diameter. Below this diameter, no linear relation prevails. All bubbles
between 2.0 and 2.4 m m in diameter were counted in one channel, and those less
than 2.0 mm were hardly detected. A comparison of bubble size distributions which
were measured at the wall by electrooptical and photographic methods (Figs. 4 and
5), indicate that the small bubbles were missing in the bubble size distributions
evaluated by the electroopticaI method with a 1.5 mm diameter sensor capillary. Fur-
thermore, the larger bubbles appeared less frequently in the distribution evaluated by
photographic measurements. Therefore, d and d s evaluated by the electrooptical
method were higher than those measured by the photographic method (Fig. 6).
Furthermore, at high superficial gas velocities (e.g., at wSG = 5.33 cm s-1, Fig. 5)
large bubbles are dispersed in the funnel and sensor capillary producing small bubbles
320 R. B u c h h o l z a n d K. Schfigerl

n
~n/Z~d

0,8

O.6

0`,

Oj

0 1 2 3 ~ 5 6 7 8
w~33cm/s d(mm)

n
T nlxd

0,4 i" """ v "~\

II i ~"
0,2 . /9

1 Z 3 4 5 6 7 8
wS6=3,2Ocm/s d(mm)

Fig. 5. Comparison of the bubble size distributions measured at the wall by photographic, electri-
cal conductivity - and electrooptical methods. Distilled water, b = 132 cm; WSL = 1.8 cm/s. II. At
high superficial gas velocities, WSG. For symbols see Figure 4

a ~s
9 9 electrooptico~ method
o A photogrophical method
WsL= 1,8 c m / s
porous plate 50,urn
distiUed water

Fig. 6. Comparison of mean bubble diameters, d, and

Sauter bubble diameters, ds, measured by the electro-
optical and photographic methods at the wall. Porous
plate, 50 t~m. Distilled water, wSL = 1.8 cm/s. d, d s,
9 , 9 electrooptical method; o, z~: photographic method
 7
Fable 1. Comparison of m e t h o d s
7"

Highly ;election
~queous viscous Fermenta- vith regard
aaodel model ti on Bubble size o t he
dethod ned ia media media distribution ?osition ,ubble size ~emarks

?lash ces yes yes broa d o n l y at tarrow bubbl e 3 ~

)hotography 0.1 to the wall ize: no selection,
20 m m )road bubbl e size:
arge bubbles are
uppressed
O
Slectrical ces yes yes a broad independent 1es ocal gas hold o O

:onductivity 0.6 to rabbles with tp and bubbl e

;ensor with 20 m m I < 0.6 m m are ,elocity can also
:wo sensors letected unsatis- ~e measured
t~
'actorily o~ N"
Slectro- (es no yes b for a single independent 7es ocal gas hold N

)ptical sensor sensor lepends on the lp can also be

capillary: liameter of neasured
t~
na rrow :apillary o=.
~.
Light scat- r yes yes a medium independent res ocal gas hold
:ering glass 1 to 20 m m mbbles with lp and bubbl e
!iber sensor I < 1 m m are ,elocity can also P~
~ith tw o lOt d e t e c t e d ,e measured

tSensor probe can be sterilized

)Sterilization problems due to removal of m e d i u m f rom the reactor and feeding it ba c k
i
"3
5"
322 R. Buchholz and K. Schfigerl

All of them can be applied to aqueous solutions. The electrical conductivity sensor
in unsuitable for organic liquids. Except for electrooptical sensors, sensors can be
applied to highly viscous media. Because of the non-spherical shape of large bubbles,
the double sensor-sensors d o not yield enough information to reliably estimate bubble
volumes and shapes. This also applies for pure liquids or solutions with antifoam agents.
Four or five sensor-sensors are necessary when large bubbles are present.
Except for the flash photography, the methods are not restricted to a given column
position. Difficulties arise due to the different sensitivity of the methods for bubbles
with unlike sizes. Flash photography can handle the broadest bubble size distribution,
however, it suppresses large bubbles.
The electrical conductivity and/or glass fiber sensor suppresses bubbles with d <
0.6 mm and/or d 1 <~ mm. Electrooptical sensors can only handle bubbles with narrow
size distributions. If the diameter d k of the sensor capillary, is much larger than d,
then the bubbles will not be detected. When d is much larger than dK, the bubbles
will be dispersed and a false bubble size distribution will result. Thus, dK must be care-
fully adjusted to d in case of narrow distributions. For broad distributions, two or
more sensor capillaries are needed.
Electrical conductivity probes and glass fiber sensors may be sterilized if they are
appropriately constructed. In the authors laboratory, the position dependence of bub-
ble size distributions during fermentation is measured by four electrical conductivity
sensors simultaneously. The sterilization problem of the electrooptical sensors is a dif-
ficult one, because the fermentation medium must be removed from the reaction and
is usually fed back into it. Hence, the entire sampling by-pass system must be sterilized.
No simple method is known, by which it is possible to reliably measure a very broad
bubble size distribution in bubble columns.

Acknowledgements. The authors gratefully acknowledge the financial support given by the Ministry
of Research and Technology in Bonn, Federal Republic of Germany.

Symbols

(L = length, T = time, M = mass)

d bubble diameter L
J mean bubble diameter L
dK inside diameter of the sensor capillary L
longitudinal distance between start and stop sensors L
Z~lB pierced length of bubble L
t time T
zxt1 length of square-wave signal at start sensor T
zxt2 length of square-wave signal at stop sensor T
•t12 time delay between start and stop signals T
VB bubble volume L3
VB bubble velocity LT-1
wSG superficial gas velocity LT-1
Bubble Column Bioreactors II 323

References

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and R. Scheunemann, eds., DFVLR p. 137
Buchhoiz, R. (1979). Dissertation, Technical University Hannover
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301-313
Burgess, J.M., Calderbank, P.H. (1975). Chem. Eng. Sci. 30,743
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Calderbank, P.H. (1960). Int. Symp. Destillation, Brighton (Inst. Chem. Eng.) p. 51
Calderbank, P.H. (1967. Mass transfer in fermentation equipment. In: Biochemistry
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Calderbank, P.H., Pereira, J. (1977). Chem. Eng. Sci. 32, 1427
Delhaye, J.M. (1969). Proc. 11th Nat. ASME/AIChE Heat Transfer Conf. on Two
Phase Flow Instrumentations. Minneapolis, Minn. p. 58
Geake, J.E., Smaller, C. (1975). The Chemical Engineer, p. 301 May
J ekat, H. (1975). Dissertation, Technical University Munich
Pilhofer, T., Miller, H.D. (1972). Chem. Ing. Techn. 44, 295
Pilhofer, T. (1974). Chem. Ing. Techn. 46,913 (MS 149/74)
Sauter, J. (1928). Forsch. Arb. Geb. Ing. 2-8,312
Todtenhaupt, E.K. (1971). Chem. Ing. Techn. 43,337
Trice, V.G., Roger, W.A., Rushton, J.H. (1956). AIChE-Journal 2, 205
Vermeulen, T., Williams, G.M., Langlois, G.E. (1955). Chem. Eng. Progress 51, 85

Received October 26, 1978