UPSTREAM & DOWSTREAM

PROCESS TECHNOLOGY

Sasmita Sabat M.Sc., Ph.D. Microbiology

Department of Biotechnology
UPSTREAM & DOWSTREAM PROCESS
TECHNOLOGY

MEMBRANE SEPARATION - MODULES

Sasmita Sabat
Assistant Professor, Department of Biotechnology
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Membrane modules
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE MODULES
Membrane systems exploit the inherent properties of
• Equipment high selectivity, high surface-area-per-
•Reservoir for the feed unit-volume, and their potential for
•High pressure pump controlling the level of contact and/or
•Membrane module
mixing between two phases.
•High pressure regulators and
•Gauges
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
Criteria for membrane module design
•High membrane surface-volume ratio
•An adequate Structural support to allow thin membrane
to withstand
•A low pressure drop on the concentrate side of the
membrane to maintain the driving force for permeation
•Turbulence on the concentrate side to dissipate
concentration polarization
• Provision for back-flushing
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE MODULES
Chemical Resistance and Compatibility
• Operating pH range
• Compatibility with solutes
• Binding of solutes
• Compatibility with solvents
• Compatibility with adhesives
• Compatibility with cleaning agents
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE MODULES
Characteristics of broth for membrane separation process
Broth characteristics such as
• Small particle size
• wide particle-size distribution
• High solids compressibility
• Increasing viscosity as the suspension becomes more concentrated
• The introduction of non-Newtonian fluid behaviour
• Minimal density differences between the solid and liquid phases
Factors/Requirements for separation
• Selectivity and high separation efficiency
• Relative size
• Electrostatic charge
• Other non-covalent interactions
• Diffusivity
• Shape
• Volatility
• Solubility
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE MODULES
• High permeate flux rate
• Mechanical/physical strngth to withstand the high pressure operation without
elongation of the pores.
• Durability and consistency of performance over prolonged periods
• Resistance to corrosion
• Low cost and should be readily available.
• High hydraulic permeability towards solvent
• Appropriate sieving property (i.e. semi-permeability)
• Chemical stability
• Thermal stability
• Manufacturing reproducibility
• Amenability to fabrication and packaging
• Stability freedom from fouling and packaging
And must be …
\Easy to clean
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE MODULES
Organic vs. Inorganic membrane
• They are inexpensive • They can withstand higher
• They were developed before transmembrane pressure
inorganic membranes • They are generally more durable
• They are light (although ceramic and glass
membranes can be quite brittle and
• They are flexible and can easily be hence susceptible to breakage)
cast or moulded into various
shapes and sizes • They are generally resistant to a
wider variety of chemical substances
• Certain membrane types (e.g. e.g. acids, alkali and solvents
hollow fibres) can only be
prepared with organic polymers • They can easily be cleaned and
sterilised
• A wide range of membrane
chemistry is available
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE MODULES
Membrane module configuration
•Flat sheet membrane/Plate-and-Frame module
•Spiral-wound membrane
•Tubular membrane
•Hollow fibre module
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE MODULES
Stirred Cells
• Uses flat sheet element
• Used for research and small scale manufacturing
• Provides defined and controlled operating conditions
• Mainly dead ended operations
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE MODULES
Flat sheet membrane/Plate-and-Frame module
•Earliest design, simple filtration technology, easy to fabricate
and use.
• Flat sheets have dimension of 1m by 1m by 200 um
•It is fabricated by stacking several flat sheets of membrane as a
multilayer sandwich in a plate and frame filter press type
arrangement.
•Module can withstand high pressure drop upto 30-40kg/cm2.
•It has very small membrane area per unit separator volume.
** Disadvantage is small membrane area per unit separator
volume.
** Leaks caused by many gasket seals,
** Costly
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE MODULES
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE MODULES
Spiral-wound membrane
•The module is fabricated by winding a membrane sheet in double layer into spiral.
• Consists of flat sheet membranes separated by spacers for the flow of feed and
permeate is wound around central perforated collection tube to form module.
•The feed is passed axially or spirally over the outer side of the double layer.
•The permeate move spirally a pipe located at the centre of the spiral.
•The spiral wound membrane has a high surface to volume ratio.
•It requires clean feeds free from particulate matter.
• This is inserted into a pressure vessel.
• Typical spiral wound module is 0.1 to 0.3 m in dia and 3 m long.
• Packaging density : 200-800 m 2 /m 3 volume
• Resistance to fouling : Moderate
• Ease of cleaning : Fair
• Relative cost : Low
• Main application : RO, UF
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE MODULES
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE MODULES
Tubular membrane
•The module is also called shell and tube heat exchanger module as it contains
5-7 smaller tubes nested inside larger tube
•Consist of tubes of membranes bounded at each end to a common header and
packed in a perforated shell.
•The feed enters to the lumen of the tubes the permeate pass through the
wall (shell) while the retentate passes out at the other end of the tube.
• •It is used in high viscous feed or feed containing particulate matter need to
be concentrated.
• Can be used for high viscosity feeds
• 0.5 – 5.0 cm in dia and upto 6 m in length
• Packaging density : 30-200 m 2/ m3 volume
• Resistance to fouling : Very good
• Ease to clean : Excellent
• Relative cost : High
• Generally used for ultrafiltration
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE MODULES

Source: http://synderfiltration.com/learning-center/articles/
module-configurations-process/tubular-membranes/
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE MODULES
Hollow fibre module
1. Formed in two basic geometrics
(i) Shell –side feed design :Closed bundle of hollow-fibre is
contained in a pressure vessel.
• The other end of the tube has epoxy resin.
• Hollow tubes are 100um and 150-200 um
(ii) Bore –side feed type: The hollow fibre are open at both
the ends and the fluid is usually circulated through the
bore of the fibres.
2. Packaging density : 500-9000 m 2 / m 3 volume
3. Resistance to fouling : Poor
4. Ease to clean : poor
5. Relative Cost : Low
6. Application : RO, UF, Dialysis
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE MODULES

: Stephen A. Rackley. Membrane separation systems InCarbon Capture and Storage (Second Edition)
2017, Pages 187-225
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Membrane Modules
REFERENCES
Text Book:
• Upstream and Downstream Processing of Bioproducts, R.
Puvanakrishnan, S. Sivasubramanian, T. Hemalatha. MJP
Publisher, New Delhi, · 2019
Reference Books:
• Downstream Process Technology, N K Prasad, PHI Learning
Private Ltd, New Delhi, 2010.
UPSTREAM & DOWSTREAM PROCESS
TECHNOLOGY

MEMBRANE SEPARATION

Sasmita Sabat
Department of Biotechnology
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
Introduction and types of membrane process
• A membrane is a selective barrier that permits the
separation of certain species in a fluid by combination of
sieving and diffusion mechanisms. Membranes can
separate particles and molecules and over a wide particle
size range and molecular weights.

Source: https://marketresearch.biz/report/membrane-separation-market/
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
Advantages of membrane separation
• Low energy alternative
• Thermally liable materials can be concentrated
•Chemical and mechanical stress on the product are
minimal
• Single step product concentration and purification can
be achieved
• Easily scale up.
Goal
• The goal is to allow one component of a mixture to
permeate the membrane freely, while hindering
permeation of other components.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
Membrane filtration can be classified into;
•Microfiltration (MF),
•Ultrafiltration (UF),
•Nanofiltration (NF) and
•Hyper filtration / Reverse osmosis (RO).
•Each of the filtration membranes has its own characteristic pore
size and separation limits

•Microfiltration (commonly abbreviated to MF) is a type of

physical filtration process where a contaminated fluid is passed
through a special pore sized membrane to separate
microorganisms and suspended particles less than 2μm in size
from process liquid.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
• Basis of separation is on pore size on the membrane
•It is used for the separation of high molecular weight
product like polymers, proteins and colloidal materials
from low molecular weight solute.
•Ultra filtration uses a finely porous membrane to
separate water and micro solutes from
macromolecules and colloids.
•The average pore diameter of the membrane is in the
range of 100–1000˚A.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
Basic types of microfiltration membrane
• Depth filters
•Capture particle at constrictions of the interior surface
of the pores or by adsorption into the pore wall.
•Particle trapped within the membrane of the filter
reduces pressure resulting in fall of permeability,
which requires membrane replacement.
•Larger holding capacity before fouling due to larger
surface area available for collection of solute
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
• Screen filters
• Collect particles whose size is larger than the pore
diameter on the surface of the membrane.
•Collected particle on the surface of membrane,
require recirculation.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
Classification of Membranes
• Synthetic membranes display a broad range in their
physical structure and the material they are made
from. They can be classified based on their
morphology.
• In symmetric membrane, the material is same with
uniform porosity.
• In asymmetric membrane, a thin skin of lower
porosity over a symmetric support acts as a
membrane.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
Schematic Diagrams of the principal types of membranes
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
Isotropic Membranes
• Microporous Membranes
• A microporous membrane is very similar in structure and
function to a conventional filter.
• It has a rigid, highly voided structure with randomly
distributed, interconnected pores.
• These pores differ from those in a conventional filter by
being extremely small, on the order of 0.01 to 10 μm in
diameter.
• All particles larger than the largest pores are completely
rejected by the membrane.
• Particles smaller than the largest pores, but larger than
the smallest pores are partially rejected, according to the
pore size distribution of the membrane.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
• Nonporous, Dense Membranes
• Nonporous, dense membranes consist of a dense film
through which permeates are transported by diffusion
under the driving force of a pressure, concentration, or
electrical potential gradient.
• The separation of various components of a mixture is
related directly to their relative transport rate within the
membrane, which is determined by their diffusivity and
solubility in the membrane material.
• Thus, nonporous, dense membranes can separate
permeates of similar size if their concentration in the
membrane material (that is, their solubility) differs
significantly.
• Most gas separation, pervaporation, and reverse osmosis
membranes use dense membranes to perform the
separation.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
• Dense homogeneous polymer membrane
• Usually they are prepared
(i) from solution by solvent evaporation or
(ii) by extrusion of the melted polymer.
• However, dense homogeneous membranes only have
a practical usefulness when they are made from a
highly permeable polymer such as silicone.
• Commonly, the permeate flow across the membrane
is quite low, since a minimal thickness is required to
grant the membrane mechanical stability.
• Most membranes currently are porous or consist of a
dense top layer on a porous structure.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
• Electrically Charged Membranes
• Electrically charged membranes can be dense or
microporous, but are most commonly very finely
microporous, with the pore walls carrying fixed positively
or negatively charged ions.
• A membrane with fixed positively charged ions is referred
to as an anion-exchange membrane because it binds
anions in the surrounding fluid.
• Similarly, a membrane containing fixed negatively charged
ions is called a cation-exchange membrane.
• Separation with charged membranes is achieved mainly by
exclusion of ions of the same charge as the fixed ions of
the membrane structure, and to a much lesser extent by
the pore size.
• The separation is affected by the charge and
concentration of the ions in solution.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
• Anisotropic Membranes
• The transport rate of a species through a membrane is inversely proportional to the
membrane thickness.
• High transport rates are desirable in membrane separation processes for economic
reasons; therefore, the membrane should be as thin as possible.
• Conventional film fabrication technology limits manufacture of mechanically strong,
defect-free films to about 20 μm thickness.
• Anisotropic membranes consist of an extremely thin surface layer supported on a
much thicker, porous substructure
• The surface layer and its substructure may be formed in a single operation or
separately.
• In composite membranes, the layers are usually made from different polymers.
• The separation properties and permeation rates of the membrane are determined
exclusively by the surface layer; the substructure functions as a mechanical
support.
• The advantages of the higher fluxes provided by anisotropic membranes are so
great that almost all commercial processes use such membranes.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
Ceramic, Metal and Liquid Membranes
•The membrane materials are organic polymers and,
infact, the vast majority of membranes used
commercially are polymer-based.
•Ceramic membranes, a special class of micro-porous
membranes are being used in ultra-filtration and
microfiltration applications for which solvent
resistance and thermal stability are required.
•Dense metal membranes particularly palladium
membranes are being considered for the separation
of hydrogen from gas mixtures and supported liquid
films are being developed for carrier-facilitated
transport processes.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
Membrane materials
In general there are three fundamentally different
categories of membrane materials:
•Organic (polymeric),
•Inorganic (ceramic) materials and
•Biological materials.
Organic materials are either cellulose-based or
composed of modified organic polymers.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
•Inorganic materials such as ceramics and metals are
used in industrial applications but are usually
cost-prohibitive in waste water treatment.
•Biological membrane (biomembranes) is a selective
barrier within or around a cell in a living organism.
•The bio- membrane is capable of recognising what is
necessary for the cell to receive or block for its
survival.
•These membranes cannot meet the industrial
requirements due to thermo-mechanical stability and
productivity.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
•A large majority of membranes in research and
commercial use are polymeric-based (organic
membrane) as a result of their facile processing into
viable membrane structures and the diverse
polymers available as well as the capability to
synthesise novel polymer structures.
•Recently composite membranes and inorganic
membranes have gained tremendous attention
owing to their potentially high performance, long life
time and even their availability that outweigh the
benefits/advantages of using polymeric membranes.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
•Inorganic membranes possess excellent thermal and
chemical stability in comparison to polymeric
membranes and higher antifouling property due to
the hydrophilic nature of inorganic material.
•Inorganic membranes are generally divided into four
groups:
•Glass membranes
•Ceramic membranes
•Metallic membranes
•Carbon membranes
•Zeolitic membranes
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
Membrane separation based on solute size
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
Theoretical model for membrane processes
•Capillary flow model
•The membrane is considered to be loose and
microporous capable of retaining particle larger than
10 Å.
•The flow of the feed occurs through the pores by
connective flow in another wise impermeable layer.
•A filtering or sieving type mechanism occurs where in
the solvent moves through the micro-pores.
•Passage of the larger molecules will be prevented by
the size of the micro-pores.
•E.g. Microfiltration, Ultra filtration
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
• Solution diffusion model
•The model postulates the dissolution of the
molecular species being transported in the material
of the membrane followed by molecular diffusion
across the barrier.
•Driving force is concentration gradient applied
pressure difference.
•E.g. Reverse osmosis
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
• Retention coefficient or rejection coefficient
•The separating ability of a membrane in pressure
driven process of MF, UF and RO is explained in terms
of retention or rejection coefficient which is defined
as
R=Cm–Cp/Cm
•R is the theoretical retention coefficient and Cm and
Cp are concentration of the solute at the membrane
surface and in the permeate.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
• Factors affecting the separation process

• Concentration polarisation at the membrane

surface
•Fouling of the membrane

Source: Goosen, M.F.A., Sablani, S.S., Al-Hinai, H., Al-Obeidani, S., Al-Belushi, R., and Jackson, D., Separat.
Sci. Technol., 39, 2261, 2004
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
Operational requirement of membrane
•Selectivity and high separation efficiency
•High permeate flux rate
•Mechanical/physical strength to withstand high
pressure operation without elongation of the pores
•Durability and consistency of the performance over
prolonged periods
•Resistance to corrosion
•Ease of fabrication in appropriate shape
•Low cost and readily available
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
Structure of the membrane
•RO and UF–generally two phase (A thin (0.5–10μm)
dense layer of material with micro-porous structures
at top and supported by a thick layer (50–125μm) of
relatively micro-porous material).
•Single casting–membrane of 0.1–0.2mm in thickness.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
Preparation of membranes
•Synthetic membrane
•Cellulose acetate
•Cellulose phthalate
•Polyamide
•Polyacrylo nitrile
•Polyethylene
•Poytetra fluoro ethylene
•First step–base materials is dissolved in suitable solvent
with necessary additives to give homogenous solution of
desired viscosity.
•Second step–film is cast by spreading the solution on a
clear dry glass plate or drawn as a hollow tube with
proper thickness control.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
•Third step–the cast film is maintained in a controlled
atmosphere to allow evaporation of the solvent.
•Fourth step–gelation the membrane dipped in a water
bath at controlled temperature in the range 0-40C to leach
out the solvent and the additives slowly there by creating
micropores in the membrane.
•Final step–annealing heating the membrane in a water
bath 70 -900C to allow the shrinkage of the pores formed
during gelation.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
• Preparation of cellulose acetate membrane
•Casting the solution contains cellulose acetate (25%),
acetone (45%) and formaldehyde (30%). In addition
small quantities of magnesium perchlorate (1-5g).

•Evaporation time is 2-3minutes at 20-250C gelation is

at 20C for 5 minutes and annealing 800C.
UPSTREAM & DOWSTREAM PROCESS
TECHNOLOGY

MEMBRANE SEPARATION – I
MICROFILTRATION

Sasmita Sabat
Department of Biotechnology
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MF

Milestone in the development

of Microfiltration
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MF
Characteristics of MF

• The ability of membranes to reject particles is

controlled by the pore size of 0.08 to 2μm for MF
membranes (0.002 to 0.1 μm for UF membranes as
compared to less than 0.001μ for RO membranes and
for NF membranes falls between the RO and UF and is
in the order of 0.001μ to 0.01μ).

•MF and UF membrane processes are true filtration

processes where particle separation occurs according
to their size.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MF
•In the MF and UF membrane processes the reject
stream contains fine particles plus bacteria and other
pollutants found in the feed.
•The MF, UF and NF filtration processes normally consist
of the membrane modules, a pressure pump, cartridge
filter and a raw water supply system.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MF

Microfiltration membranes are natural or synthetic

polymers
•Cellulose nitrate or acetate
•Polyvinylidene difluoride(PVDF)
•Polyamide
•Polysulfone
•Polycarbonate
•Polypropelene
•Poly tetrafluoro ethylene(PTEE)
•Inorganic materials (metal)
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MF
Criteria for selection of membrane
•Mechanical strength
•Temperature resistance
•Chemical compatibility
•Wettability
•Adsorption characteristics
•Hydrophobic characteristics
•Hydrophilic characteristics
•Permeability
•Permiselectivity
•Cost of the membrane
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MF
• Porous membrane is
divided according to their
pore diameter:
1. Microporous (dp < 2
nm),
2. Mesoporous (2 nm < dp
< 50 nm) and
3. Macroporous (dp > 50
nm)
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MF
Applications
•Pharmaceutical industry
•Concentration of fruit juice and alcoholic beverages in
the food and beverage industry
•Sterilization preparation of the chemical industry
•Cold sterilization of beer in fermentation industry
•Concentration of biomass
•Separation of soluble product
•Harvesting microbial cell from the fermented broth
•Separation of Blood cell and plasma from whole cell
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MF
Disadvantages
•Fouling of the membrane due to deposition of solids
on the membrane which necessitate spurging of the
unit periodically.
•Limiting pressure drop, the membrane need to be
cleaned or replaced.
•Concentrated slurry or dry cake cannot be obtained.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MF
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
MEMBRANE SEPARATION
REFERENCES
Text Book:
• Upstream and Downstream Processing of Bioproducts, R.
Puvanakrishnan, S. Sivasubramanian, T. Hemalatha. MJP
Publisher, New Delhi, · 2019
Reference Books:
• Downstream Process Technology, N K Prasad, PHI Learning
Private Ltd, New Delhi, 2010.
UPSTREAM & DOWSTREAM PROCESS
TECHNOLOGY

MEMBRANE SEPARATION – II
ULTRAFILTRATION

Sasmita Sabat
Assistant Professor, Department of Biotechnology
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
UF
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
UF
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
UF
• The permeate flux equation in dead end in microfiltration
flow model

•Where Ji is solvent flux in kg/(sm2),

–ΔP the pressure difference in Pa,
–μ is the viscosity of the solvent in Pas,
–Rm is the membrane resistance in m/kg and
- Rc is the cake resistance in m/kg, which is increases with
time due to cake build up.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
UF
• The permeate flux equation in cross flow microfiltration
flow model

• Where, Ji is solvent flux in kg/(sm2),

•Mass transfer coefficient
•D is the rejected species diffusion coefficient
•δ is the thickness of the concentration polarization
boundary level
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
UF
Comparison of advantages and disadvantages of in-line and cross-flow microfiltration
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
UF
Ultra filtration Membranes
•Ultra filtration membranes are usually anisotropic
structures made by the Loeb–Sourirajan process.
•They have a finely porous surface layer or skin
supported on a much more open microporous
substrate.
•The finely porous surface layer performs the
separation; the microporous substrate provides
mechanical strength.
•The membranes discriminate between dissolved
macromolecules of different sizes.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
UF
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
UF
UF-Criteria for selection of membrane
•Mechanical strength
•Temperature resistance
•Chemical compatibility
•Hydrophobic/Hydrophilic characteristics
•Permeability
•Permi selectivity
•Cost of the membrane material
•Manufacturing process
•Pore size smaller than microfiltration membrane
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
UF
UF membrane materials
• Polymers Polyvinylidiene fluoride (PVDF) Current standards (> 85% solutions)
• Polyether sulfone (PES)
• Polysulfone (PS) PVDF
• Polyacrylonitrile (PAN) • Chemical stability
• Polyethylene (PE) •Mechanical strength
• Polypropylene (PP) •Durability
• Polyvinyl chloride (PVC) PES
• Ceramic membranes •Hydrophilicity
UF membrane expected properties •Low polymer cost
▪ Mechanical strength •History (early ‘90s)
▪ Hydrophilicity
▪ Durability
▪ Chemical stability
▪ Low polymer cost
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
UF

Source: Membrane technology and applications by Richard W.

Baker.—2nd ed. 2004.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
UF
FACTORS AFFECTING THE PERFORMANCE OF
ULTRAFILTRATION
A. Flow velocity
• Critical for liquids containing emulsions or suspensions
• Higher flow velocity reduces the fouling
B. Operating pressure
• Permeate rate is directly proportional to the applied
pressure
C. Operating temperature
• Permeate rates increase with increasing temperature
D. Flow type
• Dead end configurations: batch processes with low
suspended solids
• Cross flow configurations: continuous operations
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
UF
UF-Choice of membrane
•Spirally wound
•Capillary module
• Hollow fibre
• Tubular
• Plate and frame
Effectiveness of the Membrane
1. Physical properties of the membrane
• Permeability
• Thickness
2. Process and system variables
• Feed consumption
• Feed concentration
• System pressure
• Velocity
• Temperature
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
UF

Cheese Production Method

Source: Membrane technology and applications / Richard W. Baker.—2nd ed.

2004.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
UF

APPLICATION OF
ULTRAFILTRATION
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
UF
UF IN FOOD INDUSTRY
• Recovery of whey protein and concentration of skim
milk in dairy industry
• Clarification and bacteria removal of wine
• Recover valuable products from soya whey and other
dilute waste streams
• Purification of fermentation solution, and clarification
and concentration of fruit juice
• Concentration of gelatin
• Recovery of sugar from sugary water
• Fractionation and concentration of egg albumin,
proteins, extracts such as vanilla, lemon, peel extract,
etc., and animal, fish and vegetable oils
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
UF
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
UF
REFERENCES
Text Book:
• Upstream and Downstream Processing of Bioproducts, R.
Puvanakrishnan, S. Sivasubramanian, T. Hemalatha. MJP
Publisher, New Delhi, · 2019
Reference Books:
• Downstream Process Technology, N K Prasad, PHI Learning
Private Ltd, New Delhi, 2010.
UPSTREAM & DOWSTREAM PROCESS
TECHNOLOGY

MEMBRANE SEPARATION – II
REVERSE OSMOSIS
NANOFILTRATION

Sasmita Sabat
Department of Biotechnology
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
RO
• Reverse osmosis is a process for desalting water
using membranes that are permeable to water but
essentially impermeable to salt.
•Pressurized water containing dissolved salts contacts
the feed side of the membrane; water depleted of salt
is withdrawn as a low pressure permeate.
•The nomenclature is based on the fact that the
direction of normal osmotic flow of a solvent across a
semi- permeable membrane is reversed due to an
applied pressure which is greater than the osmotic
pressure of the liquid feed.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
RO
• Currently approximately (more than) 15 million
m3/day of water are desalted by reverse osmosis,
providing approximately more than 2% of the world’s
drinking water supply.
•Half of this capacity is installed in the Middle East and
other desert regions to produce municipal water from
wastewater, brackish groundwater or the sea.
•The remainder is installed in the United States, Europe
and Japan principally to produce ultra pure industrial
water.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
RO
• The solvent molecules pass from a region of lower
concentration to a region of higher concentration.
•The driving force for the flow of the solvent across the
membrane Is the chemical potential on the two sides
of the membrane.
•This continues until the fluid pressure in the
concentrated solution is sufficiently high to prevent
the passage of further solvent molecules.
•At equilibrium the chemical potential on both sides of
the membrane are equal and the fluid pressure is
called the osmotic pressure of the solution.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
RO
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
RO
• The RO promotes flow of the solvent from more
concentrated solution to less concentrated solution.
•Since osmotic pressure opposes the desired direction
of flow pressure sufficient to nullify the osmotic
pressure and overcome the resistance of the
membrane is applied on the concentrated solution
side.
•The RO membrane is anisotropic with a skin of active
membrane of 0.2-0.5 micron.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
RO
Membranes and Materials
•A number of membrane materials and membrane
preparation techniques have been used to make
reverse osmosis membranes.
•The target of much of the early work was sea water
desalination (approximately 3.5 wt % salt) which
requires membranes with salt rejections of greater
than 99.3% to produce an acceptable permeate
containing less than 500ppm salt.
•Early membranes could only meet this target
performance when operated at very high pressures
upto 100 bar.
•As membrane performance has improved the
operating pressure has dropped to 50–60 bar.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
RO
1. Cellulosic Membranes
•Cellulose acetate was the first high-performance reverse
osmosis membrane material discovered.
•The flux and rejection of cellulose acetate membranes
have now been surpassed by interfacial composite
membranes.
2. Interfacial Composite Membranes
•Since the discovery by Cadotte and his co-workers that
high-flux,high-rejection reverse osmosis membranes can
be made by interfacial polymerization membranes made
by this method have become the industry standard.
•Interfacial composite membranes have significantly higher
salt rejections and fluxes than cellulose acetate
membranes.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
RO
• Reverse Osmosis Membrane Categories
•Reverse osmosis membranes can be grouped into
three main categories:
•Seawater and brackish water desalination membranes
operated with 0.5–5wt% salt solutions at pressures of
10–60 bar.
•Low pressure nanofiltration membranes operated
with 200–5000pp malt solutions at pressures of 5–10
bar.
•Hyperfiltration membranes used to separate solutes
from organic solvent solutions.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
RO
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
RO
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
RO
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
RO
• Membrane selectivity
•Multivalent ions are retained better than mono valent
ions. Although the absolute values of the salt
rejection vary over a wide range. The ranking for the
different salts is the same for all membranes.
•In general the order of rejection of ions by reverse
osmosis membranes is as shown below.
•Fe3+ >Ni2+≈ Cu2+ > Mg2+ > Ca2+ > Na+ > K+
•For anions:
•PO43−> SO42−> HCO3−> Br−> Cl−> NO3− ˷ F−
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
RO
• Membrane modules
•Spiral-wound modules
•Hollow fibre modules

• Applications
•Approximately half of the reverse osmosis systems
currently installed are used for desalinating brackish water
or seawater.
•Another 40% are producing ultrapure water for the
electronics, pharmaceutical and power generation
industries.
•The remainder is used in small niche applications such as
pollution control and food processing.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
RO
Contd….
1. Concentration of enzymes
2. Concenration of sugar
3. Dialysis
4. Blood oxygenation
5. Organic solvent separation
6. Waste water treatment
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
RO
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
NF
Nanofiltration
• Nanofiltration membranes are intermediate between
ultrafiltration membranes – clearly microporous – and reverse
osmosis membranes – clearly dense films.
• Nanofiltration membranes have high rejections for the di- and
trisaccharides, sucrose and raffinose, with molecular diameters
of 10-13 Å, but freely pass the monosaccharide fructose with a
molecular diameter of about 5-6 Å.
• Nanofiltration is a membrane filtration-based method that uses
nanometer sized cylindrical through-pores that pass through
the membrane.
• The permeation properties of nanofiltration membrane can be
neutral, positively charged, and negatively charged.
• The neutral nanofiltration membrane rejects the various salts in
proportion to molecular size, so the order of rejection is simply
Na2SO4> CaCl2> NaCl
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
NF
• The anionic nanofiltration membrane has positive
groups attached to the polymer backbone. These
positive charges repel positive cations, particularly
divalent cations such as Ca2+, while attracting negative
anions, particularly divalent anions such as SO42−.
• The result is an order of salt rejection
CaCl2>NaCl>Na2SO4
• The cationic nanofiltration membrane has negative
groups attached to the polymer backbone. These
negative charges repel negative anions, such as SO42−,
while attracting positive cations, particularly divalent
cations such as Ca2+
• The result is an order of salt rejection
Na2SO4> NaCl > CaCl2
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
NF
Advantages
• One of the main advantages of nanofiltration as a
method of softening water is that during the process
of retaining calcium and magnesium ions while
passing smaller hydrated monovalent ions, filtration is
performed without adding extra sodium ions, as used
in ion exchangers.
• Many separation processes do not operate at room
temperature (Ex: distillation), which greatly increases
the cost of the process when continuous heating or
cooling is applied.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
NF
Disadvantages
• The membrane pores sizes are limited to only
nanometers
• The cost and maintenance of the membranes
• Repairs and replacement of membranes is dependent
on total dissolved solids, flow rate and components of
the feed.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
NF
Applications
• Industrial applications of nanofiltration are quite common in
• the food and dairy sector
• in chemical processing
• in the pulp and paper industry, and
• in textiles, although the chief application continues to be in the
treatment of fresh, process and waste waters
• The original uses for nanofiltration were water treatment and in
particular water softening.
• Fine chemistry and Pharmaceuticals
a. Non-thermal solvent recovery and management
b. Room temperature solvent exchange
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
NF
• Oil and Petroleum chemistry
a. Removal of tar components in feed
b. Purification of gas condensates
• Bulk Chemistry
a. Product Polishing
b. Continuous recovery of homogeneous catalysts
• Natural Essential Oils and similar products
a. Fractionation of crude extracts
b. Enrichment of natural compounds Gentle Separations
• Medicine
a. Able to extract amino acids and lipids from blood and
another cell culture.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
NF
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Membrane Separation
REFERENCES
Text Book:
• Upstream and Downstream Processing of Bioproducts, R.
Puvanakrishnan, S. Sivasubramanian, T. Hemalatha. MJP
Publisher, New Delhi, · 2019
Reference Books:
• Downstream Process Technology, N K Prasad, PHI Learning
Private Ltd, New Delhi, 2010.
UPSTREAM & DOWSTREAM PROCESS
TECHNOLOGY

Tangential Flow Filtration

Sasmita Sabat
Assistant Professor, Department of Biotechnology
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
TFF
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
TFF
Membrane Processes are becoming popular because
they are considered “Green” Technology- No
chemicals are used in the process.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
TFF
Tangential Flow Filtration (TFF) is a rapid and efficient
method for separation and purification of biomolecules.
•It can be applied to a wide range of biological fields such
as;
•Immunology
•Protein chemistry
•Molecular biology
•Biochemistry
•Microbiology
•TFF can be used to concentrate and desalt sample
solutions ranging in volume from 10mL to thousands of
litres.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
TFF
Why to use TFF ????
•TFF is easy to set up and use
•TFF is fast and efficient
•Perform two steps with one system
•TFF can be scaled-up or scaled-down
•TFF is economical
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
TFF
What Can Tangential Flow Filtration Do
•Concentrate and desalt proteins and peptides.
•Concentrate and desalt nucleic acids.
•Recover & purify antibodies or recombinant proteins from cell
culture media.
•Recover and purify plasmid DNA from cell lysates or
chromosomal DNA from whole blood.
•Fractionate dilute protein mixtures.
•Clarify cell lysates or tissue homogenates.
•De-pyrogenate (remove endotoxin) water, buffers, and media
solutions.
•Prepare samples prior to column chromatography.
•Harvest cells.
•Recover or remove viruses.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
TFF
What is Tangential Flow Filtration?
• Membrane filtration is a separation technique widely used
in the life science laboratory. There are two main
membrane filtration modes which can use either
microfiltration or ultra-filtration membranes:
•Direct Flow Filtration (DFF) also known as ”dead-end”
filtration, applies the feed stream perpendicular to the
membrane face and attempts to pass 100% of the fluid
through the membrane and
•Tangential Flow Filtration (TFF) also known as cross flow
filtration where the feed stream passes parallel to the
membrane face as one portion passes through the
membrane (permeate) while the remainder (retentate) is
re-circulated back to the feed reservoir.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
TFF

surface that can cause fouling.

(B) The TFF process prevents the rapid decline in flux rate seen in direct flow
filtration allowing a
greater volume to be processed per unit area of membrane surface.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
TFF
Applications
•The primary applications for TFF are concentration
•Diafiltration (desalting and buffer exchange
•Fractionation of macromolecules from small
biomolecules.
•In addition it can be used for clarification and removal
of cells as well as cellular debris from fermentation or
cell culture broths.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
TFF
Process Variables in Tangential Flow Filtration
•Two of the important variables involved in all
tangential flow devices are transmembrane pressure
(TMP) and crossflow velocity (CF).
•The transmembrane pressure is the force that drives
fluid through the membrane carrying along the
permeable molecules.
•The cross flow velocity is the rate of the solution flow
through the feed channel and across the membrane.
It provides the force that sweeps away molecules
that can foul the membrane and restrict filtrate flow.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
TFF
Tangential Flow Device Assembly
•Tangential flow filtration systems typically require a
TFF device (capsule, cassette and holder, hollowfiber
module, etc.) with a pump (peristaltic or equivalent),
tubing, valves or clamps, one or more pressure
gauges, and a sample reservoir.
•Pressure gauges are typically installed at the feed,
retentate, and filtrate ports in development and
process TFF system.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
TFF
How to Choose the Proper TFF System
•Define the purpose of the TFF process
•Choose the membrane molecular weight cut-off
•Choose the flow channel configuration
•Screen channel configuration
•Suspended screen channel configuration
•Open channel configuration
•Determine the required membrane area for the
application
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
TFF
Flow channel configuration
•Screen channel configuration: It is used with a clean, filtered
(0.2µm) solution (no particles or aggregates that can get
trapped in the screen). A woven separator in the channel
creates gentle turbulence along the membrane surface
minimizing membrane fouling.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
TFF
•Suspended screen channel configuration: It has a
more open structure in the retentate channel that
provides better performance when highly viscous
fluids (for example: serum) or particle-laden
solutions are being used. It can also be used to
concentrate cells or clarify cell or fermentation
broths.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
TFF
•Open channel configuration: It is used in the same
applications as the suspended screen channel. This
configuration has no screen in the feed channel.
Instead it uses spacers to define the channel height.
Devices may be available in several channel heights.
Typically a channel height between 0.5 and 1.0 mm
is used force harvest applications. This structure
minimizes cell disruption and maximizes recovery of
intact cells after concentration.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
TFF
Operation of a TFF system consists of the following steps
•Rinse the TFF device before use to remove the storage agent.
•Establish the Normalized Water Permeability (NWP) of the
membrane to establish a baseline for the device performance.
(This step is not necessary but strongly recommended if the
device will be cleaned and reused.)
•Condition system with the sample buffer (Conditioning helps
remove air from the system adjust system temperature and
prevent possible precipitation or denaturation of biomolecules
resulting from contact with flushing solution).
•Process the sample (concentration and/ or diafiltration or
fractionation).
•Clean;determine cleaning efficiency.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
TFF
REFERENCES
Text Book:
• Upstream and Downstream Processing of Bioproducts, R.
Puvanakrishnan, S. Sivasubramanian, T. Hemalatha. MJP
Publisher, New Delhi, · 2019
Reference Books:
• Downstream Process Technology, N K Prasad, PHI Learning
Private Ltd, New Delhi, 2010.
UPSTREAM & DOWSTREAM PROCESS
TECHNOLOGY

Membrane separation
Problems in membrane separation and control measures
•Concentration polarization process
•Membrane fouling and factors affecting

Sasmita Sabat
Department of Biotechnology
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Concentration polarization
Factors affecting the separation process
•Concentration polarisation at the membrane surface
• Fouling of the membrane
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Concentration polarization
Fouling
Factors causing fouling
–Slime formation
–Microbial growth
–Deposition of macromolecules (UF)
–Colloid deposition
–Physical compaction of the membrane
•It is irreversible need to replace the membrane
•Selection of the membrane (hydrophilic membrane less
prone to fouling of proteins)
•Pre-treatment of the feed (pH, precipitation)
•Cleaning of membrane with chemicals
•Back-flushing with permeate
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Concentration polarization
Concentration polarisation at the membrane surface
•In membrane separation processes, a gas or liquid
mixture contacts the feed side of the membrane, and
a permeate enriched in one of the components of the
mixture is withdrawn from the downstream side of
the membrane.
•Because the feed mixture components permeate the
membrane at differentiates, there is a gradual
build-up in the concentration of non-permeating or
slowly permeating components in the feed as the
more permeable components pass through the
membrane.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Concentration polarization
•A layer is formed near the surface of the membrane,
where by the solution immediately adjacent to the
membrane surface becomes depleted in the
permeating solute on the feed side of the membrane,
and its concentration is lower than that in the bulk
fluid.
•On the other hand, the concentration of the
non-permeating component increases at the
membrane surface.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Concentration polarization
• Concentration polarisation at the membrane surface
•A concentration gradient is formed in the fluid
adjacent to the membrane surface.
•This phenomenon is known as concentration
polarization and it serves to reduce the permeating
component's concentration difference across the
membrane, there by lowering its flux and the
membrane selectivity.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Concentration polarization
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Concentration polarization
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Concentration Polarization
Types of Fouling
1. Particulate deposition:
• Standard blocking: macromolecules are uniformly
deposited on pore walls
• Complete blocking: membrane pore is completely
sealed by a macromolecule
•Cake formation: accumulated particles or
macromolecules form a fouling layer on the
membrane surface, in UF this is also known as a gel
layer
• Intermediate blocking: when macromolecules deposit
into pores or onto already blocked pores, contributing
to cake formation
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Concentration Polarization
2. Scaling:
As a result of concentration polarization at the membrane surface,
increased ion concentrations may exceed solubility thresholds and
precipitate on the membrane surface.
These inorganic salt deposits can block pores causing flux decline,
membrane degradation and loss of production. The formation of scale
is highly dependent on factors affecting both solubility and
concentration polarization including pH, temperature, flow velocity
and permeation rate.
Scale is caused by precipitation of dissolved metal salts in the feed
water on the membrane surface
Organic fouling is the attachment of materials such as oil or grease
onto the membrane surface.
3. Biofouling
• Microorganisms will adhere to the membrane surface forming a gel
layer – known as biofilm.
• The film increases the resistance to flow, acting as an additional
barrier to permeation.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Concentration Polarization
Membrane fouling: The nature of fouling in low pressure membrane
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Concnetration Polarization
Factors affecting fouling
• Membrane properties:
• Pore size
• Hydrophobicity
• Pore size distribution and
• Membrane material
• Solution properties:
• Solid (particle) concentration,
• Particle size and
• Nature of components.
• Operating conditions:
• pH
• Temperature
• Flow rate and
• Pressure
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Concentration Polarization
• Membrane fouling
•Pretreatment Techniques for Controlling Fouling
•Coagulation
•Adsorption
•Bio-filtration
•Magnetic Ion Exchange Resins
•Integrated Pre-treatment Systems
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Concentration Polarization
• There are three main factors that contribute to
membrane fouling:
•Feed stream properties: This includes characteristics
such as concentration, pH and ionic strength,
dissolved solid content, suspended solid content,etc.
•Membrane material and its physical-chemical
properties: Membrane poresize, porosity and pore
distribution, and membrane surface charge and
hydrophobicity in certain pH and ionic strength
conditions can contribute to fouling.
•Processing variables: Transmembrane pressure,
temperature, and cross-flow velocity have a big
impact on membrane fouling.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Concentration Polarization
• Approaches can be taken to minimize membrane
fouling
•Optimize pH and ionic strength of the feed solution to
minimize the adsorption or deposition of the feed
materials.
•Select an appropriate pre-filtration procedure or other
means to remove large molecules, since the presence
of larger molecules or particles could cause as the rich
hindrance to the passage of smaller molecules
through the membrane.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Concentration Polarization
•Select a membrane with an optimum pore size to
result in good separation performance as well as
optimized permeate flux.
•Optimize the operating conditions. This includes
increasing transmembrane pressure to maximize flux
without introducing more fouling potential.
•Increase the cross-flow velocity, which generally
results in an improvement in permeate flux.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Concentration Polarization
How do we know if our facility has a problem with
membrane fouling?
•Problems with membrane fouling can emerge
gradually or suddenly.
•Early on, facilities will often see symptoms such as
rising energy costs and decreased membrane flux.
•As foulants continue to accumulate on the
membrane surface, even greater pressure is
needed to force water through, which can
ultimately lead to irreparable damage to the
membrane and other system components.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Concentration Polarization
• Concentration polarization is the accumulation of
rejected particles, especially during microfiltration
and ultrafiltration, to an extent that transport to the
membrane surface becomes limited.
•High flux through the membrane can cause the
rejected particles to accumulate on the surface of the
membrane.
•Concentration polarization reduces the permeability
of the solvent and can lead to a limiting flux, in which
an increase in pressure does not correspond to arise
influx.
•Reducing concentration polarization leads to higher
limiting flux and lower fouling tendency.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Concentration Polarization
• Depending on the solute and process conditions,
three effects can reduce flux in concentration
polarization
•Rising resistance, which leads to lower flux,
•Increasing osmotic pressure due to retained
macromolecules, which reduces transmembrane
pressure,
•Changing physio-chemical properties, such as viscosity
within the membrane boundary layer
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Concentration Polarisation
• Acid cleaning agents such as Hydrochloric,
Phosphoric, or Citric acids effectively remove common
scaling compounds.
•With Cellulose Acetate membranes the pH of the
solution should not go below 2.0 or else hydrolysis of
the membrane will occur.
•Oxalic acid is particularly effective for removing iron
deposits.
•Acids such as Citric acid are not very effective with
Calcium, Magnesium, or Barium Sulphate scale; in this
case a chelating agent such as Ethylene Diamine
Tetraacetic Acid (EDTA) may be used.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Concentration Polarization
• To remove bacteria, silt or precipitates from the
membrane, alkalis combined with surfactant cleaners
are often used.
•Laundry detergents containing enzyme additives are
useful for removing bio-foulants and some organic
foulants.
•Most large membrane module producers now
distribute formulated products, which are a mixture
of cleaning compounds.
•These products are designed for various common feed
waters and often provide a better solution to
membrane cleaning than devising a cleaning solution
for a specific feed.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Concentration Polarization
• Sterilization of a membrane system is also required to
control bacterial growth.
•For Cellulose Acetate membranes, chlorination of the
feed water is sufficient to control bacteria.
•Feed water to polyamide or interfacial composite
membranes need not be sterile, because these
membranes are usually fairly resistant to biological
attack.
•Periodic shock disinfection using Formaldehyde,
Peroxide or Peracetic acid solutions as part of a
regular cleaning schedule is usually enough to prevent
biofouling.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Concentration Polaratization
There are various ways to reduce this fouling such as
•Periodic pulsing of feed
•Periodic pulsing filtrate (backwashing)
•Increasing shear at by rotating membrane
•Vibrating membrane (VSEP technology)
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
NF
• A common method to clean the membrane system is
to just reverse the flow pattern:
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
NF
• A common method to clean the membrane system is
to just reverse the flow pattern

Vibrating shear to prevent fouling

UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Membrane Separation
REFERENCES
Text Book:
• Upstream and Downstream Processing of Bioproducts, R.
Puvanakrishnan, S. Sivasubramanian, T. Hemalatha. MJP
Publisher, New Delhi, · 2019
Reference Books:
• Downstream Process Technology, N K Prasad, PHI Learning
Private Ltd, New Delhi, 2010.
• https://www.slideshare.net/hazz12/ultrafiltration-865339
78
UPSTREAM & DOWSTREAM PROCESS
TECHNOLOGY

PRECIPITATION

Sasmita Sabat
Department of Biotechnology
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
PRECIPITATION
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Precipitation
Involves the conversion of soluble solutes into
insoluble solids which can be separated subsequently
from liquid by physical separation methods.
Recovery of bulk protein
Preliminary stages of downstream processing
Induced by: salt/organic solvent addition, changing of
pH value to alter the nature of the solution.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Precipitation
• Proteins precipitate as a result of insolubility.
• Proteins solubility is determined by the interactions
between surface regions with surrounding water
molecules.
• Several factors affecting the protein solubility are
considered important: Structure, Size, Charge and
Solvent
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Precipitation
Structure and Size
In the native state, a protein in an aqueous environment
assumes a structure that:
❖ Minimizes the contact of the hydrophobic amino acid
residues with the water solvent molecules.
❖ Maximises the contact of the polar and charged residues
with the water.
Charge
❖ The net charge of protein has a direct bearing upon the
proteins solubility.
❖ The solubility of a protein increases as its net charge
increases, a result of greater interaction with dipolar
molecules.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Precipitation
Solvent
The solvent affects the solubility of proteins primarily
through two parameters: Hydrophobicity and ionic
strength.
The major forces to stabilize a protein in its native state:
- Hydrogen bonding, Van der Waals interactions,
Solvophobic interactions (driven forces of folding
protein).
In aqueous solution these forces tend to push :
1) Hydrophobic residues into the interior of the
protein
2) The polar and charged residues on the proteins
surface.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Precipitation
• Hydrophobicity
• Single phase solutions of water and monohydric alcohols
denaturation at room temperature can be avoided at
low temperature.
• Denaturing efficiency increases:
methanol-ethanol-propanol etc
• Longer alkyl chain of alcohols: Effective binding to polar
groups
• Weak intraprotein hydrophobic: interactions denature
• When the temperature is low- the monohydric alcohols
compete for the water of hydration on the protein and
cause the protein molecules to approach more closely so
that the van der waals interactions lead to aggregation.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Precipitation
Ionic strength
Can have both effects: Solubilizing (Salting in) and
Precipitation (Salting Out)

Salting in: The addition of small quantities of neutral

salts to a protein solution which often increases
protein solubility.

Salting out: Addition of salts above an optimal level

which leads to destabilization of protein in solution
eventually promotes protein precipitation.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Precipitation
Water molecules bind to the surface of the protein
molecule because of association of charged and polar
groups and immobilization of non polar molecules.
The surface of proteins usually contains a significant
fraction of non-polar atoms. The forces acting on a
protein lead to the achievement of a minimum Gibbs
free energy---------native protein only marginally
stable and can be destabilized by relatively small
environmental changes.
The hydration layers are thought to promote solubility
of the protein by maintaining a distance between the
surfaces of the protein molecules.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Precipitation
• The solubility of protein increased as its net charge
increases as a result of greater interaction with water
molecules.
• A repulsive reaction between protein molecules of
same charge further increases solubility.
• A simple way to vary the charge on a protein is by pH
alterations.
• Isolelectric pH -------pH of the solution in which a
protein has net charge ZERO
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Precipitation
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Precipitation
• Precipitation is an important method in purification
of proteins and is used in bulk recovery of proteins.
•The primary advantages of precipitation are that
it is relatively inexpensive
can be carried out with simple equipment can be done
continuously
leads to a form of the protein that is often stable in
long-term storage.
•Since precipitation is quite tolerant of various
impurities including nucleic acids and lipids it is used
early (primary method of isolation or recovery and
concentration) in many bio-separation processes.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Precipitation
• Proteins in their natural condition generally exhibit an
negative charge on the surface and attract positive
ions to form the Stern layer of counter ion close to
the protein surface.
• The stern layer is surrounded by a diffuse
Gouy-Chapman layer of mobile counter ions.
•Stability of the protein-electrolyte colloid is due to the
balance between attractive and repulsive forces
between colloidal particles not allowing them to form
aggregates.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Precipitation
Isoelectric Precipitation
•Isoelectric point where the net charge on the protein is
zero.
•Proteins have no net charge at their pI values and the
electrostatic repulsions between protein molecules are
minimum at these conditions results minimum solubility
of the proteins at its pI.
•This is different for different proteins.
• A convenient method when fractionating a protein
mixture
• At isoelectric pH the proteins will be: Neutral pH
Solubility decreases, Buffering capacity decreases and
precipitation increases
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Precipitation
•The pH should be adjusted above the highest pI or
below the lowest pI of all the proteins present.

•The pH change is brought to the nearest pI where the

precipitation is allowed.
•It is positively charged at low pH and negatively
charged at high pH.
Advantages: When the acids are added to cause
precipitation : mineral acids (Cheap and easily
available are acceptable in protein food industries.
Gelatin which is very hydrophilic protein does not
precipitate at its isoelectric point in solvents with low
ionic strength.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
PRecipitation
•The intermediate pH at which a protein molecule has
a net charge of zero is called the isoelectric point of
that protein.
•In general, the net charge on the protein either
positive or negative can interact with water molecules
meaning that it is more likely for a protein molecule to
dissociate itself from other protein molecules thus
more soluble.
•As a result protein is the least soluble when the pH of
the solution is at its isoelectric point.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Precipitation
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Precipitation
• Isoelectric focusing is a separation technique which
separates peptides according to their isoelectric point
or how acidic and basic their residues are.
•A gel with a pH gradient is used as the medium. The pH
gradient is made by adding polyampholytes which are
multi-charged polymers with different pI into the gel.
•Then the sample is put onto the gel and a voltage is
applied.
•The proteins will move along the gel until they reach
their isoelectric points. In other words each protein will
move until it reaches a position in the gel at which the
pH is equal to the pI of the protein.
•A protein band that forms at a given pH can then be
removed and analyzed further. This process can
successfully separate proteins that have a difference in
net charge greater than or equal to 1.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Precipitation
Addition of organic solvents
• Several organic solvents have been used to precipitate
proteins including alcohols, acetone and ether,
dioxan,2-methoxyethanol,etc.
• Alcohols are widely used in industry.
• Cohn process: One of the most important processes
utilizing alcohol to precipitate the proteins to purify
therapeutic proteins from human plasma.
• Uses ethanol at temperatures below 0oC to minimize
denaturation by the organic solvent.
• Manipulated variables: pH, ionic strength and ethanol
concentration.
• Ionic strength kept at low leading to salting effect.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Precipitation
Addition of anionic polymers
• Several nonionic polymers have been used to
precipitate proteins : dextran, poly vinyl pyrrolidone,
poly propylene glycol, polyethylene glycol.
• They are soluble in water.
• PEG:
Solutions of PEG up to 20 % w/v can be used without
viscosity becoming a problem
PEG with molecular weights above 4000 found to be
most effective
Protein destabilization in PEG does not occur until the
temperature is significantly higher than room
temperature (40oC).
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Precipitation
•Reducing the effective amount of water available for
the solvation of protein and excluding the proteins
from part of the solution

•Higher concentration of PEG required to precipitate

proteins of low molecular weight

•Protein precipitated by non-ionic polymers can be

processed by ion-exchange resins directly without any
intermediate step to remove the precipitating agent.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Precipitation
• Polyethylene glycol is a non denaturing water-soluble
polymer whose ability to precipitate protein from
aqueous solution can be qualitatively understood in
terms of an excluded volume mechanism.
•The increment in PEG concentration required to effect
a given reduction insolubility is unique for a given
protein-polymer pair being insensitive to solution
conditions and primarily dependent on the size of the
protein and polymer.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Precipitation
Precipitation by metal ions
•Polyvalent metal ion cause protein precipitation
•Based on mode of action:-------
•Ions such as divalent manganese ,iron, cobalt, nickel,
copper, zinc and cadmium bind strongly to carboxylic
acids and to the nitrogenous groups such as amine
and heterocyclic group.
•Ions such as divalent calcium, barium, magnesium and
lead bind to carboxylic acids but not significantly to
nitrogenous ligand
•Ions such as monovalent silver, mercury and lead bind
strongly to sulfhydryl groups.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Precipitation
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Precipitation
• https://www.youtube.com/watch?v=i_6y6Z5UvwE
SDS

• https://www.youtube.com/watch?v=gnhUh6qVD5Y
Precipitation
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Precipitation
Protein precipitation by addition of salts
•Salt induced precipitation
•Increase in protein solubility by increase in salt
concentration in the ionic strength range of zero to
about 0.5M at given pH and temperature.

Source:
https://www.quora.com/How-does-the-salting-in-phenomenon-of-proteins-work-to-
make-protein-soluble
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Precipitation
• Precipitation of proteins by adding salt is called salting-out.
•The added salt remove water from hydrated protein exposing
hydrophobic regions to interact inter-molecularly resulting
their aggregation.
•Charge on anion or cation influence their ability to bring about
the precipitation of protein and formulated a series of ions
referred as lysotropic series

https://mypetitedress-shorts.blogspot.com/1978/06/cation-and-anion-charge-chart.html
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Precipitation

Hofmeister observed that

the charge on the anions as
well as cations influenced
their ability to bring about
precipitation of proteins and
formulated a series of ions.

Source:https://www.pnas.org/content/116/46/23029/tab-figures-data
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Precipitation
Salting in
Salting out

Source: GyörgyHegyiet. al., Introduction to Practical Biochemistry

UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Precipitation
Advantages of ammonium salt
✔ Its solubility is very high (approximately 4M in pure water)
and varies very little in the range of 0o to 30oC.
✔ The density of its saturated solution is 1.235gcm-3,
enough below the density of protein aggregates (approx
1.29gcm-3) to allow centrifugation.
✔ Protein precipitated are often very stable for years.
✔ Proteolysis and bacterial action are precented in
concentrated ammonium sulphate solutions.
❑ Disadvantages: Cannot be used above pH 8 because of the
buffering action of ammonia.
Sodium citrate is highly soluble and is an good alternative to
ammonium sulphate precipitation above pH 8.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Precipitation
Salting in Salting out
1. At low concentrations added 1. At high concentrations added
salt usually increases the salt lowers the solubility of
solubility of charged macromolecules because it
competes for the solvent
macromolecules because (water) needed to solvate the
the salt screens out macromolecules.
charge-charge interactions.
2. So high Salt removes the
2. So low salt prevents solvation sphere from the
aggregation and therefore protein molecules and they
“precipitation or crashing” come out of solution.
UPSTREAM & DOWSTREAM PROCESS TECHNOLOGY
Membrane Separation
REFERENCES
Text Book:
• Upstream and Downstream Processing of Bioproducts, R.
Puvanakrishnan, S. Sivasubramanian, T. Hemalatha. MJP Publisher,
New Delhi, · 2019
Reference Books:
• Downstream Process Technology, N K Prasad, PHI Learning Private
Ltd, New Delhi, 2010.
• https://www.slideserve.com/lea/product-purification-precipitation

• https://www.zmchdahod.org/pdf/college/Reactions_of_Protein-01-
11-2018.pdf
THANK YOU

Dr. Sasmita Sabat

[email protected]
+91 80 26721983 Extn 351