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Settler

Under Guidance of Prof. Pushpavanam

Submitted by:

Rohit Singh

5th Year Dual Degree Student

Chemical Engineering

In this report we describe the work carried out in the period from

August 2015 to March 2016. The work has been carried out in a

systematic manner to help logically analyze the results. First we

discuss the sedimentation in a batch vessel which is vertical and

then is inclined. Then we extend this to a continuous system. The

analysis is done using Fluent as the platform for all simulations.

INTRODUCTION

Suspension mammalian cell culture processes for the production

of biologic therapeutics (antibodies) are operated in a variety of

modes such as: batch, fed-batch, chemostat, and perfusion.

In perfusion culture, a continuous supply of fresh media is fed

into the bioreactor while growth-inhibitory by-products are

constantly removed. The advantages of perfusion cell culture, in

particular, include high volumetric productivities via higher cell

densities than the other modes and better cell physiology

control. Perfusion is also advantageous when product stability is

a concern as the product can be harvested continuously as

opposed to waiting for the duration of a batch or fed-batch

process.

Hybridoma is a hybrid cell used as the basis for the production of

antibodies in large amounts for diagnostic or therapeutic use.

Early studies in batch hybridoma culturing showed that

monoclonal antibody production is proportional to the

number of viable cells in the culture [1,2] .Nonviable or

dead cells do not lose significant amounts of antibody and,

consequently, contribute negligibly to the productivity of a culture

[3]. Thus, high productivity of monoclonal antibody in suspension

culture can be attained not by maximal cell growth rate but by

maintaining high viable cell concentrations.

The accumulation of nonviable cells and the removal of viable

cells along with nonviable cells limit the culture productivity.

Limitations in culture productivity may be overcome by

continuously removing only nonviable cells from the reactor while

selectively retaining all viable cells in the reactor, provided that a

method may be developed for accomplishing a selective

separation of these subpopulations.

Cell retention devices are employed in perfusion processes to

separate the cells from the product-containing supernatant

in the bioreactor. Such cell-separating devices include

centrifuges, hollow-fiber filters, hydrocyclones, gravity settlers,

spin filters and ultrasonic separators.

High viable cell concentrations in suspension bioreactors are

currently achieved by using various cell retention or recycle

devices. Filter devices can be detrimental to long-term culture

productivity because the cells are subjected to excessive shear

forces, resulting in higher cell death rates. Inclined plate settlers

have various advantages over traditional settling devices. It can

be operated continuously unlike the batch sedimentation devices.

Inclined settlers generally do not contain any internal moving

parts and accomplish the separation by utilizing the force of

gravity within an environment of minimal shear stress.

It is expected that the desired selective cell separation may be

accomplished by exploiting the different sedimentation

velocities of viable and nonviable cells by using inclined

sedimentation. Larger cells are removed from suspension by

settling onto the upward-facing surfaces of the settler, where

they form thin sediment layers that slide down to be collected at

the bottom of the vessel.

Batt et al [6] performed experiments and concluded that at high

dilution rates ,over 95% of the viable cells could be partitioned to

the bottom of the settler while over 50 % of the nonviable cells

are removed through the top of the settler. This successful

separation is due to the significantly larger size of the viable cells

than the nonviable ones.

THE BOYCOTT EFFECT & INCLINED PLATE SETTLERS

The benefit of inclined sedimentation is that cells need to settle

only a distance on the order of the narrow spacing between the

inclined walls of the settler in order to be removed from

suspension, rather than a distance on the order of the settler

height as in vertical sedimentation. In order to explain the

Boycott effect, Ponder (1925) and Nakamura and Kuroda (1937)

independently proposed an analytical model to predict the

sedimentation rate for a monodisperse suspension in a parallelplate container. This model is called the PNK theory. [4-5]

dh

dt

= -Vo(1 +

h

b sinA

velocity in a vertical container of the same dimensions. It is clear

that the settling rate is enhanced by a factor (h/b) sin A. That is,

the settling rate will be improved for a larger ratio of the

suspension length to the width, b, or by a larger inclination angle.

Kinosita (1949) experimentally observed convection currents in

an inclined tube, which generated a strong vortex. He found that

some particles trapped in the vortex move up to 100 times faster

than those sedimentation particles. Hill et al. (1977) proposed a

continuum model, and he was followed by Acrivos and

Herbolzheimer (1979), Borhan and Acrivos (1988), and Kapoor

and Acrivos (1995). In these studies, Acrivos and his coworkers

paid attention to the modeling of the sediment layer on the

upward-facing surface and the way it drains towards the bottom

of the container. Their results agree better with the experimental

data than the PNK theory does.

b: plate spacing;

n: number of plates;

w: plate width;

l: plate length;

: settler angle

more than a half-century ago. This theory states that the

volumetric production rate of clarified fluid from an inclined

channel due to particle sedimentation is equal to the vertical

settling velocity of the particles multiplied by the horizontal

projected area of the channel surface available for

sedimentation [4-5].

S(u)= uw (Lsin + b cos )

(1)

of particles with settling velocity u ,w is the width of the settler,

L is the length of the settler, b is the spacing.

The scale-up of an inclined sedimentation device can be

accomplished by either increasing the dimensions of a single

plate, which can result in long cell residence times and

cumbersome physical sizes, or by utilizing multiple plates.

These multiple plate designs, however, share two common

problems. First, a single pump removes the overflow for many

plates, causing inevitable flow distribution problems. For

example, it is impossible to maintain identical flow rates up each

of the channels. The second, more serious problem is that cells

which have settled and are sliding off the top channels will be reentrained in the up flow through lower channels. In this manner,

viable cells will not be swiftly returned to the bioreactor upon

settling, and the residence time of cells of the settler can be

increased dramatically.

Selective removal of nonviable cells can be achieved by

adjusting the settler overflow rate so that the residence time of

the suspension in the settler is in between the settling times of

two subpopulations of different sizes.

For a given cell culture growing at a particular rate, the degree of

cell separation was found to depend on the overflow rate through

the settler, which determines the residence time of suspended

cells in the sedimentation channel.

An efficient settler will rapidly return all cells to the

uninhibited by the low settler temperatures and uncontrolled

nutrient conditions. The length of time that cells spend in the

settler is therefore critical to considerations of settler

dynamics.

This is a proprietary patent-pending design (shown in Figures 2 and 3) of Compact cell settler developed

by Sudhin Biopharma Co. It exploits the centrifugal forces on cells (in the cell culture medium from the

bioreactor) entering tangentially near the top of cylindrical portion of the cyclone assembly. Cells pushed

against the spiral cylindrical walls settle down vertically to reach the conical bottom of the cyclone

housing and its internal inclined surfaces, which enhances the cell settling as discovered by Boycott

(1920) with blood cells and confirmed by Batt et al., (1990) for hybridoma cells and Searles et al., (1994)

for CHO cells.

% of the cell concentration in the

bioreactor sample.

1. Cylindrical portion of cyclone holding vessel;

2. Conical bottom section of cyclone vessel;

3. Top headplate sealed with o-rings on cylcone;

4. Central top outlet port welded into headplate;

5. Screws attaching the lid to the cyclone

6. Tangential port near the top for inlet

of cell culture medium from

bioreactor;

7. Vertical spiral plate with constant

spacing between successive rings;

8. Conical spiral surface (a single continuous surface

or several angled plates) welded to the vertical

spiral plate; and,

9. Bottom outlet port for returning settled cells back

to the bioreactor.*

This prototype was constructed and housed inside a

cyclone of 12 inches outer diameter and 12 inches

total height as shown in Figure 2. It is a vertical

spiral plate turning 5 full circles, with several

angled plates welded at the bottom. In the initial

experiments with this device at a perfusion rate of

5 liters/day harvested from yeast Pichia cells

growing in 5-liter bioreactor contained only 5 10

counter, the yeast cells exiting in the

harvest were significantly smaller than

those in the bioreactor.

Spiral Settler device

Settler device

PROBLEM STATEMENT

To find the optimal parameters and design specifications for the

above described cell settler.

For the design optimization, some of the suggestions given to us

were to change the height of cylindrical portion keeping the

volume constant and seeing its effect.

Simulating the above problem and geometry was a challenging

task. For simulating the above geometry, we first needed to

understand the dynamics of the system. After understanding the

system and developing an appropriate model for a simpler system,

we can extrapolate it to the much complex system.

and inclined channels

1. Vertical bimodal sedimentation

The geometry considered for the study was a vertical cuboidal

sedimentation column, 100 cm tall and has a square cross-section

with 5 cm side. Two types of particles were considered for the

simulations. Group 1: 137 microns, initial volume fraction 0.01,

density=2440 kg/m3 and Group 2: 198 microns, initial volume

fraction 0.04, density = 2990 kg/m3 .Both types of particles were

studied individually.

The suspending medium (fluid) consisted of a mixture of Union

Carbide UCON oils and Monsanto HB40 hydrogenated terphenyl oil

which had a Newtonian fluid with density = 992 kg/m 3 and viscosity

= 0.0677 kg / m-s. This viscosity is sixty seven times higher than

that of water. The maximum particle volume fraction (packing

fraction) is 0.53 [3] This choice of parameters is based on the

experimental study in the literature and is used to validate the

numerical code developed.

The geometry being simulated is shown in Fig.4. All the simulations

are performed in 2-D as we can find a plane of symmetry (XY

160 respectively.

Geometry

Meshed Geometry

Number of y cells = 160

column

Model and Solver Specifications

Euler Euler Granular Model is used for carrying out the

simulations. The key features of the model are now described.

To take into account the sliding of particles on the inclined surface,

a slip boundary condition is used for particle-wall interaction.

No slip condition is used for fluid-wall interaction.

The coefficient of restitution between particle-particle is set as

1 (collisions are elastic) since there exists a thin liquid film covering

the particle surface.

The coefficient of restitution between particle-wall is set as 0

(inelastic collision). This implies that the particles are assumed to

be deposited when they come in contact with the wall.

The particle viscosity and particle collision stress is modeled

by the kinetic theory of granular flows. Solids phase properties

become functions of the solids phase volume fraction and the

granular temperature which accounts for velocity fluctuations

owing to particle-to-particle collisions. [9,15]

A second order upwind scheme is used to solve the momentum

equations for the convective terms.

A transient coupled solver was used to solve the momentum,

continuity, granular temperature and energy equations. Time step

size for the simulation was determined on a case-by-case basis and

was chosen to be as large as possible such that the solution

converged. The time step size used varies from 5 x10-3 s to 2x10-2 s

for most simulations.

For convergence of the solution, the residuals were monitored .The

criterion used for convergence was residuals had to reach 10 -3 at

every time step. The time step was chosen such that for each time

step the numerical scheme converges in 5-10 iterations.

Results

The experimental data was obtained from Davis R.H et al (1982)

The sedimentation of polydispere suspensions in vessels

having inclined wall. Simulation of sedimentation in a vertical

column was done in Fluent 14.1.

Fig.5 shows the comparison between experimental data and

simulated data for the two particles. It can be seen that the

simulation is able to predict the experimental behavior for both

types of particles.

100

95

Experimental group 2

90

Height (cm)

Experimental Group 1

Simulated Group 2

85

Simulated Group 1

80

75

0 50 100 150 200 250300 350400 450

Time (s)

results

sedimentation

For these simulations the vertical height was kept constant at 40

cm and the cross-section of the geometry was a square with side 5

cm. The particles were assumed to be monodisperse with a size of

137 microns and density of 2420 kg/m 3. Initial volume fraction of

particles was taken as 0.1 .The maximum particle volume fraction

(packing fraction) = 0.53

The fluid properties considered chosen were density = 992 kg/m 3

and viscosity = 0.0677 kg / m-s. This channel was analyzed for

three different inclinations; 350, 200 and 00.

Meshing

The solution obtained depends on the grid size chosen. The final

solution which has numerically converged should be independent

of the grid. The solution changed initially when we went from

coarse mesh to fine mesh. But after a critical mesh size, the

solution becomes independent of the grid size.

The final mesh size used has a maximum face size of 1 mm. This

gave a total of 19138 elements and 18590 nodes.

Another mesh of Maximum face size 1.5mm having 8292 elements

and 7948 nodes was also used. No significant difference (less than

5%) was observed in the results obtained by these two meshes.

Hence all simulations were carried out with the finer mesh.

Sedimentation rate was found to increase as we used a finer mesh.

This also gave a realistic solution (closer to experimental results)

while using a very fine mesh. In particular it was necessary for the

meshing near the boundary (walls) to be very fine (1 mm) as

sedimentation is a boundary phenomenon.

Models and solvers used here are same as that used for the

vertical bimodal sedimentation case. Meshing is different for the

inclined channels to take into account the formation of sediment

layer and its sliding on the boundary.

Geometry

Case 1: Inclination 350

3: Inclination 00

Case

Fig.6 represents the three cases being modeled. The angle of

inclination is measured from the vertical.

Results

The experimental data for the inclined channels were obtained

from [9]. Simulations of inclined columns were done in Fluent 14.1.

Fig.7 shows the comparison between experimental data and

simulation predictions for the case of the vertical channel and two

different inclinations of 20 and 35 degrees. A good agreement

between the experimental and simulated data is observed. This

validates the model.

The location where the concentration was 50% of the initial

concentration was taken to be the interface height (i.e., cell

fraction =0.05).

It must be emphasized that while the experiments were carried out

by using polydisperse particles around a mean diameter, the

simulations are based on a monodisperse suspension being used.

For the broad size distribution, the concentration just above the top

of suspension is zero, whereas, for the narrower distributions, it is

not. The effect of the polydispersity is to spread out the particles in

the upper portion of the settling vessel. [11-12]

In sedimentation experiments for monodisperse particles, Laux H.

et al (1997) encountered a comparatively sharp interface below

which the particle volume fraction equals its initial value and above

which it is virtually zero. In the computed solution, however,

numerical diffusion tends to smear the interface such that it

does not appear sharp any longer [14]. It seemed natural to define

the interface position by the contour line for half of the initial

particle volume fraction [15].

The height settled in 600 seconds is 7.3 cm for the vertical

geometry as compared to 21.5 cm for 35 0 inclined geometry. The

350 inclined settler gave a much higher settling as compared to the

vertical channel. This shows that the CFD model used with the

different assumptions is able to capture the Boycott Effect and can

be applied to a continuous system.

40

0 Experimental

35

0 Simulation

30

20 Experimental

Height (cm) 25

20 Simulation

20

35 Experimental

15

35 simulation

10

0

100

200

300

400

500

600

700

Time (s)

The settling of the particles induces a velocity field in the liquid

phase. This is readily predicted by the CFD simulations. The

velocity flow field thus determined helps us in understanding the

cause behind increased settling in inclined channels. Kohara et al.

[10] previously used a fluid mechanical model to develop a better

understanding of the enhanced settling of mammalian cells within

open-ended inclined spaces. They found that the convective flow

within the plates was 20 times the Stokes velocity of the cells.

Figure 8 (a) depicts the velocity contours and 8 (b) the velocity

vectors in a channel inclined at 35 degrees to the vertical. Figure 9

is a zoomed version of the channel which shows the recirculation at

the top and bottom. This clearly confirms the circulation pattern in

the inclined channel.

(zoomed in)

pattern

For the particle size chosen (137 m), the Stokes settling velocity

of particle is = 0.0215 cm/s and is obtained from

g ( p f ) D

v s=

18

frac)4.6 This comes by following Richardson & Zakis method

where

Vhindered = Vterminal (1volume fraction)n , n=4.6 in this case .This corresponds to the

domain where particle Reynolds number Re p is less than 0.2 . Using

this, the hindered Settling velocity of particles is obtained as 0.013

cm/s

We now discuss the y velocity component in the system which

determines the settling velocity. This is depicted in Figures 10 and

11 for a vertical and inclined channel respectively.

It can be seen that for the vertical system the y-velocity of the

particles are comparable to the hindered sttling velocity predicted

by Richardson and Zaki correlation. However for an inclined

channel, the y-velocity of particles trapped in the vortex ranges

from -0.24 to -1.73 cm/s (at time = 100 s). This settling velocity

is 20-150 times that calculated by Richardson & Zakis method

see Fig.11.

For 350 inclined system,

indicates the height at which velocity profile is calculated)

The vortex comes down and becomes smaller when the particles

settle. Also, their respective velocities decrease with time.

Comparing the Y-velocity at 100 s and 300 s, we can see that the

maximum Y-velocity decreased from 1.6 cm/s to 1.2 cm/s. This was

expected as rate of sedimentation decreases with time.

The flow field in the particle-containing region was considerably

more complex and several interesting phenomena were observed.

At the start of the experiment, a rapid circulating motion developed

throughout most of the suspension. The observed circulation was

due to continuity which requires that there be no net flow across a

plane of constant x; hence the upward flow of the suspension

caused by the motion of the clear-fluid layer must be accompanied

by a corresponding downward flow elsewhere in the suspension.

The particles close to the clear-fluid interface were

observed to rise rapidly, come to an abrupt stop at the top of

immediately adjacent to that where the particles were rising. This

can be inferred from Figure 9.

Time =

100 s

Time =

300 s

and 300 s (Double arrow indicates the height at which velocity

profile is calculated)

velocity of particles as calculated by Richardson & Zakis method is

0.013 cm/s. The Hindered settling velocity of particles as

calculated by Richardson & Zakis method and the settling

velocities of particles obtained from simulations are in agreement.

settling

To study the effect of impact of inclined surface on settling we

study three different geometries each have a different ratio of the

vertical and inclined portions. These are depicted in Fig.9 .The

three cases are such that

Case 1: More straight (30 cm), less inclined (15 cm)

Case 2: Equal straight, Equal inclined

Case 3: Less straight (8.5 cm), more inclined (42 cm)

Angle of inclination is 450

Case 1

Case 2

Case 3

All the three geometries have a constant volume and vertical

height of 40 cm.

were:

Case 1: 29.0 cm

Case 2: 25.4 cm

Case 3: 19.2 cm

The clear interface is marked at the position where the particle

volume fraction reduces to half of the initial concentration. The

interface heights are measured from the bottom.

Comparing above results with the ones stated earlier, Fig. 10

contains the simulation predictions of the interface height versus

time for the different geometries.

It is clear from the above graph that the larger the fraction of the

inclined surface, the more is the settling. This was expected as

there is more area available for the particles to settle and slide.

As the batch system simulation predictions were in line with the

experimental data, it is fair to assume that the models and the way

the simulation is set up are accurate. We now discuss the behavior

of the continuous flow sedimentation system.

For the validation of model for the continuous system, The

experimental data was obtained from Batt BC, Davis RH, Kompala

DS. Inclined sedimentation for selective retention of viable

hybridomas in a continuous suspension bioreactor.

Biotechnol Prog. 1990;6:458464.

The continuous separation of nonviable hybridoma cells (8 Microns)

from viable hybridoma cells (13 Microns) by using a narrow

rectangular channel that is inclined from the vertical was

investigated experimentally. It was found that at high dilution rates

through the chemostat, over 95% of the viable cells could be

partitioned to the bottom of the settler while over 50% of the

nonviable cells are removed through the top of the settler. This

successful separation is due to the significantly larger size of the

viable hybridomas than the nonviable ones.

The effectiveness of the settler in selectively retaining viable

hybridomas in the bioreactor while permitting the removal of

nonviable hybridomas was shown to depend on the flow rate

through the settler.

The cell density was not determined exactly, but a crude neutral

buoyancy measurement indicated that it is approximately 1.06

g/cm3.

was made of glass and had the same rectangular dimensions of 5

cm in width and a 0.5 cm separation between the two inclined

surfaces. One sedimentation channel had a length of 37 cm, while

the other was 23 cm long.

Particles considered for the simulation are nonviable hybridoma

cells (8 Microns) and viable hybridoma cells (13 Microns) . The

fluid properties considered were density = 998 kg/m 3 and viscosity

= 0.001 kg / m-s. The maximum particle volume fraction (packing

fraction) = 0.53

The external inclined sedimentation channel was kept at a constant

angle of 30 from the vertical, in order to provide sufficient area for

sedimentation while allowing the sediment to easily slide down the

inclined wall. Flow through the inclined settler was generated by a

peristaltic pump at the settler outlet. Cells that settle completely

from suspension are returned to the bioreactor by gravity flow of

the sediment layer. Smaller cells that do not have sufficient time to

be removed from suspension are washed out in the settler overflow

stream.

nonviable cells in the overflow stream relative to those in the

reactor. An inclined settler 37 cm in length at an angle of

inclination 300 was used in the study

stream became the reactor effluent. The feed rate was adjusted to

exceed the overflow rate slightly, with the effluent tube employed

in chemostat operation used as a level control to maintain constant

culture volume.

The final mesh size used has a Maximum face size of 0.5 mm.

Further, inflation boundary layer meshing is used wherever

required (Case 3 and 4).

sedimentation channel

(with plates) sedimentation channel

Case 1

Case 4

Case 2

Vertical

Inclined

Inclined (7 Plates)

Case 3

Inclined (4 Plates)

Outlet

Inlet

Recirculati

on

A line probe ( a yellow line) is inserted from the inlet to the outlet.

As it is a 2-dimensional simulation, the probe lies in the plane of

line for different cases.

A grid independence study was conducted to ensure that the final

solution was independent of the mesh size. The final converged

solution of Case 4 (inclined settler with 7 plates) is used for the

same.

Volume fraction vs Length

0.11

0.10

0.09

0.08

0.07

0.7 mm

0.06

0.5 mm

0.4 mm

0.05

0.04

0.03

0.02

0.01

0.00

0

10

15

20

25

Length (cm)

There is no significant difference (< 5% error) between the steady

state results obtained by using 0.5 mm and 0.4 mm mesh.

Solution Method

Pseudo transient approach is used to find a steady state solution

for continuous systems. Pseudo time step size used is 2 seconds.

scaled residuals were less than 10-4 for the converged steady state

solution. Alternatively, when the solution stops changing with time

and scaled residuals are below 10-3, the solution obtained is a

steady state solution.

Results

0.11

0.1

0.09

0.08

0.07

4 Plates

0.06

7 plates

0.05

Vertical

Inclined

0.04

0.03

0.02

0.01

0

0

10

15

20

25

Length (cm)

conditions.

The difference in curvature [between inclined(no plates) and 4-7

plates] can be attributed to the fact that it is easier to drain fluid

out as compared to the particles.This is because of the lower drag

offered to fluid as compared to particles. Particles are heavier so

they are subjected to a larger force in vertical direction. When the

particles are completely settled ( as in case of 4-7 plates), this

behaviour is not visible. But in the vertical case and the inclined

case, there is still a large proportion of particles near the outlet. So,

more fluid is being drained out of the system as compared to the

particles.

As the number of plates increases, settling increases as the

particles have a lot more area to settle. Also, the particles have to

travel a smaller vertical distance till it reaches a wall as compared

to the case without plates. So, we expect settling to increase as

more and more plates are introduced to the settler.

Case 1 (Vertical)

Case 2 (Inclined)

Case 3 (4 plates)

Case 4

(7 plates)

Figure 20: Particle Volume fraction contours at steady state under

varying --conditions

As can be seen from the contours in Fig 20, inclining the vertical

system gave a higher settling. Dark blue colored contour

represents the clear fluid. Comparing the contours for Cases 2, 3

and 4 suggests that increasing the number of inclined plate results

in increased settling of particles. Introducing plates in the inclined

system further enhanced the settling by decreasing the distance

particle has to settle before it could roll down the inclined plate and

reach the underflow (recirculation outlet).

The above inference is clearly visible from the volume fraction v/s

length plots as depicted in Fig. 19.

ANALYSIS

When dealing with micron size particles, mesh sizing becomes an

important factor. As the particles are very small (micron size) , the

meshes near the boundary should also be fine enough to capture

the particle wall interaction.

whole geometry with elements of sizes comparable to that of the

sediment. The use of inflation layer boundary mesh significantly

reduces the number of elements in the mesh as we selectively

mesh the region where the settling is predominant ,i.e., near the

wall boundary.

In particle solutions, the grid cell size should be large enough to

contain a number of particle parcels. The grid must also be fine

enough to resolve the physics of the problem. A check is made to

evaluate the infuence of the grid on the sedimentation

calculations.

4 different types of meshes were used in this analysis. The whole

idea behind this was to get a converged solution with least number

of nodes (for reducing the computational time).

Some terminologies used in Inflation meshing are defined below:

The growth rate (g) determines the relative thickness of the

adjacent inflation layer. As we move away from the face to which

the inflation control is applied, each successive layer is

approximately one growth rate thicker than the previous one. For

e.g., a growth rate of 1.2 implies that the successive layer will be

1.2 times or 20% thicker than the previous one.

The number of layers (n) control determines the actual number

of inflation boundary layers in the mesh.

First layer height (h1) determines the height of the first inflation

layer.

S. Mesh

No size

.

No. of

No. of

elements nodes

Boundar

y layer

inflation

1

2

13380

12672

Yes

Yes

1 mm

1 mm

26050

24304

No. of

Bounda

ry layer

(n)

4

4

Growt First

h rate layer

(g)

height

(h1)

1.2

0.01 mm

1.2

0.1 mm

3

4

0.7

mm

1.5

mm

25769

51950

Yes

1.2

0.02 mm

9484

18339

Yes

1.2

0.015

mm

mm (b) Zoomed in inflation boundary layer meshing

mm

=0.02 mm

=0.015 mm

RESULTS

0.12

0.1

0.08

Mesh 1

Mesh 2

0.06

Mesh 3

Mesh 4

0.04

0.02

-5

0

0

10

15

20

25

30

Length (cm)

meshes.

see the impact that boundary layer meshing has on settling

behaviour.

The first layer height of Mesh 2 is 5-10 times that of the other

meshes. As settling is a boundary (wall) layer phenomena, the first

layer height has a very significant impact on the sedimentation of

particles.

Number of elements in Mesh 2 (12672) is more than that in Mesh

4 (9484). Inspite of that , Mesh 2 underestimates the

sedimentation of particles . This is because of the larger first layer

height is Mesh 2 (0.1 mm) as compared to Mesh 2 (0.015 mm). As

the particles are very small, the meshes near the boundary should

also be fine enough to capture the particle wall interaction.

The mesh size in regions apart from the boundary has a minimal

impact on the settling rate.This can be seen by comparing the

particle volume fraction profiles obtained from Meshes 1,3 and 4.

These three meshes differ significantly in the mesh size and

consequently the number of elements. The first layer thickness of

Mesh 1,3 and 4 are 0.01, 0.02 and 0.015 respectively. Inspite of

different mesh sizes, all the three meshes gave nearly the same

solution. This suggests that the mesh sizing does not have a

significant impact in regions apart from the boundary.

Particle deposition assumption can also be used to reduce the

computational time. According to this assumption, the particles are

supposed to be deposited as soon as they reach the wall. The

particles leave the system when they come in contact with the

wall.

The problem with the above assumption is that it is only valid at

small concentrations. As long as the deposited sediment does not

influence the fluid above it, this assumption holds true. But at

higher concentrations (say 30% v/v ), this assumption may not hold

true.

Simulations

Experiments were conducted in cylindrical test tubes of diameter

4.5 cm. The fluid medium is water having density of 998 kg/m 3 and

viscosity of 0.001 kg / m-s. Calcium carbonate was used in this

sedimentation analysis. Mean diameter of calcium carbonate is 18

microns. Density of Calcium carbonate is 2800 kg/m 3.

5% w/w. This is equal to 1.8% v/v.

Conc. = 5%

80

70

60

50

Height(cm)

Vertica Settling

40

Simulated inclined

30

Simulated vertical

20

10

0

0

1000

2000

3000

4000

5000

6000

Time (sec)

results

standardized , CFD simulations are still in line with the expermental

data.

Efficiency

Description of the multiphase system :

For these simulations, the volume was kept constant at 1 cm 3. Also,

the flow rate was kept constant for system equivalence. The

particles were assumed to be monodisperse with a size of 8

microns and density of 1060 kg/m 3. Initial volume fraction of

particles was taken as 0.1 .The maximum particle volume fraction

(packing fraction) = 0.53

The fluid properties considered chosen were density = 1000 kg/m 3

and viscosity = 0.001 kg / m-s. These constant volume settlers are

then analyzed for their sedimentation efficiency.

For efficiency of a settler , we define E (Sedimentation Efficiency)

as the percentage of particles retained in the settler.

E=

100

concentration of particlesthe feed

Meshing

The solution obtained depends on the grid size chosen. The final

solution which has numerically converged should be independent

of the grid. The solution changed initially when we went from

coarse mesh to fine mesh. But after a critical mesh size, the

solution becomes independent of the grid size.

The final mesh size used has a maximum face size of 0.5 mm.

This also gave a realistic solution (closer to experimental results)

while using a very fine mesh. In particular it was necessary for the

meshing near the boundary (walls) to be very fine (0.01 mm) as

sedimentation is a boundary phenomenon.

Models and solvers used here are same as that used for the

vertical bimodal sedimentation case.

The final mesh used for computational fluid dynamics study for

different geometries are summarized below:

S. No.

Mesh

size

No. of

No. of

elements nodes

Bounda

ry layer

inflatio

n

Yes

No. of

Bounda

ry layer

(n)

5

Growt

h rate

(g)

First layer

height

(h1)

Design

1

Design

2

Design

3

Design

4

Design

5

Design

6

0.5 mm

89127

1.2

0.01 mm

0.5 mm

Yes

1.2

0.01 mm

0.5 mm

Yes

1.2

0.01 mm

0.5 mm

Yes

1.2

0.01 mm

0.5 mm

Yes

1.2

0.01 mm

0.5 mm

Yes

1.2

0.01 mm

28749

Design 1 :

h

w

Figure 27

0.5 cm x 0.1 cm .

Length (l) , height (h) and width (w) of the settler are indicated in

figure

Distance between the parallel plates is 0.5 cm

Settling Surface Area = 20 x 0.1 = 2 cm2.

Projected Area = 2 sin 30 = 1 cm2

Figure 28

= 16.13 %

Figure 29

DESIGN 2 : Inclined Cylinder

Figure 30

1 cm3 . Length of the cylinder is 25.3 cm.

Total curved surface area of the cylinder is 17.858 cm 2

Only the bottom half of the cylinder effects the settling. So only the

horizontal projection of bottom half of the cylinder is considered.

Effective Area available for sedmentation 3.6 cm2

= 31.5 %

DESIGN 3 :

CONE

Volume is 1 cm3 .

Curved Surface Area of the cone is 5.1425 cm2.

Top radius 0.59 cm Bottom Radius 0.05 cm

Height is 2.5 cm

Sin 15 ( 3.14 x 0.59 x 2.5 )

Horizontal projected Surface area is 1.19 cm2

DESIGN 4

Volume is 1 cm3 .

Surface Area is 7.878 cm2.

Top radius 0.405 cm Bottom Radius 0.02 cm

Height is 5.5 cm

Horizontal projected Surface area is 0.5136 cm 2

DESIGN 5

Volume is 1 cm3 .

Surface Area is 6.12 cm2.

Top radius 0.73 cm Bottom Radius 0.02 cm

Height is 1.75 cm

Sin 67.3 x 3.14 x 0.72 x 1.75

Horizontal projected Surface area is 1.67 cm2

DESIGN 6

Volume is 1 cm3 .

Surface Area is 7.07 cm2.

Top radius 0.95 cm Bottom Radius 0.02 cm

Height is 1.032 cm

Horizontal projected Surface area is 2.83 cm2

surface area.

1 2

V= r h

3

l= h2 +r 2

h=

3V

r2

DESIGN 6:

References

(1). Reuveny, S.; Velez, D.; Riske, F.; Macmillan, J.; Miller Production of Monoclonal Antibodies in

Culture. Deu. Biol. Stand. 1985, 60, 185-197.

(2). Birch, J.;Thompson, P.;Lambert, K.; Boraston, R. The Large Scale Cultivation of Hybridoma Cells

Producing Monoclonal Antibodies. In Large-Scale Mammalian Cell Culture; Tolbert, W.,Feder,

J.,Eds.;Academic Press: New York, 1985;

(3). Reddy, S.; Miller, W. M. Effects of Environmental Stress on Hybridoma Antibody Production and

Metabolism. AIChE Annual Meeting, November 1989.

(4). Ponder E. On sedimentation and rouleaux formation. Q J Exp Physiol. 1925;15:235252.

dans les recipients inclines. Keijo J Med. 1937;8:256296.

(6) Batt BC, Davis RH, Kompala DS. Inclined sedimentation for selective retention of viable hybridomas

in a continuous suspension bioreactor. Biotechnol Prog. 1990;6:458464.

(7) Boycott, A. E. (1920). Sedimentation of blood corpuscles, Nature, 104, 532538.

(8) Acrivos, A. and Herbolzheimer, E. (1979). Enhanced sedimentation in settling tanks with inclined

walls, J. Fluid Mech. , 92, 435457.

(9) Wu Chun-Liang, Zhan Jie-Min Eulerian simulation of sedimentation flows in vertical and inclined

vessels. Chinese Physics 2005,14, 620-628.

(10) Kohara Y, Ueda H, Suzuki E. Enhanced settling of mammalian cells in tanks

with inclined plates/simulation by fluid mechanical model and experiment. J

Chem Eng Jpn. 1995;28:703707.

(11)Acrivos, A. and Herbolzheimer, E. (1979). Enhanced sedimentation in settling

tanks with inclined walls, J. Fluid Mech. , 92, 435457.

(12) Herbolzheimer, E. and Acrivos, A. (1981). Enhanced sedimentation in

narrow tilted channel, J. Fluid Mech. ,108, 485499.

(13) Davis R.H , Herbolzheimer, E. and Acrivos, A. (1982).The sedimentation of

polydispere suspensions in vessels having inclined walls, Int. J. Multiphase Flows,8, 571

589.

(14) S.V. Patankar, Numerical Heat Transfer and Fluid Flow, Hemisphere, New

York, 1980

(15) Laux Harald , Ytrehus Tor (1997). Computer simulation and experiments on two-phase flow in

an inclined sedimentation vessel, Powder technology,94,35-49

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