1. NANOFILTRATION&ELECTROPHORESIS

2. NANOFILTERATION
• 'process intermediate between reverse osmosis and
ultra filtration that rejects molecules which have a size
in the order of one nanometer‘
• separates a range of inorganic and organic substances
from solution in a liquid
MAIN FEATURES OF NANOFILTRATION:
•Custom Build and Modular Units
•High rejection of Multi Valent ionic load such as Calcium
and Magnesium
•Stable production of purified water
•Low operating pressure compared to reverse osmosis
(RO)
•Complex Process Development Capability
•Quality Assurance and Control Management

4. PRINCIPLE
Nanofiltration is the use of
pressure to separate soluble
contaminants from water
using a semi-permeable
membrane. This is done by
diffusion through a
membrane, under pressure
differentials that are
considerable less than those
for reverse osmosis, but still
significantly greater than
those for ultrafiltration

5. PROPERTIES OF NF
• The membranes are produced in plate and frame form, spiral
wound, tubular, capillary and hollow fibre formats, from
cellulose derivatives and synthetic polymers, from inorganic
materials, ceramics especially etc
• NF withstand in very high or low pH environments,
• NF membranes tend to have a slightly charged surface, with a
negative charge at neutral pH

6. APPLICATIONS OF NANOFILTERATION:
• Industrial applications in food and dairy sector, in chemical processing, in the
pulp and paper industry, and in textiles,
• effective means of water softening, as the main hardness chemicals are
divalent.
• removal of natural organic matter from water, especially tastes, odours
and colours, in the removal of trace herbicides from large water flows.
used for the removal of residual quantities of disinfectants in drinking
water
• NF is used to concentrate whey,
• dextrose syrup and thin sugar juice are concentrated by NF, while ion
exchange brines are demineralised
• degumming of solutions in the edible oil processing sector, for continuous
cheese production, and in the production of alternative sweeteners
• the largest NF plants was installed at a petroleum refinery for the
dewaxing of oils.

7. DRAWBACK OF NANOFILTERATION:
• susceptible to fouling
• proper pretreatment, with the right membrane
material, with adequate cross-flow velocities to scour
the membrane surface clear of accumulated slime,
and by use of rotating or vibrating membrane
holders.

8. ELECTROPHORESIS:
• Electrophoresis is a separation
technique based on the migration of
ions in an electric field. The positively
charged ions migrate towards a
negative electrode and the negatively
charged ions migrate towards the
positive electrode. Ions have different
rates of migration depending on their
total charge, size, and shape and can,
therefore, be separated by this
technique
• Swedish chemist Arne Tiselius and he
was awarded Nobel prize in the year
1948 for his valuable contributions.

9. :
ELECTROPHORESIS
• BASIC PRINCIPLE AND OPERATION:
• Separation method based on differential rate of migration of
charged species in a buffer solution across which a dc electric field
is applied. An electrophoresis separation is achieved by injection of
a small volume of sample into an aqueous buffer solution which is
contained on a porous support medium like paper or semi-solid gel
or a narrow tube.
• DIFFERENT FORMS OF ELECTROPHORESIS
• Electrophoresis can be
• one-dimensional (1D) meaning one plane of separation or
• two-dimensional (2D) meaning two planes of separation
 Slab electrophoresis
 Capillary electrophoresis

10. SLAB ELECTROPHORESIS:
• In slab electrophoresis, separations are carried out on a thin
flat layer (slab) of porous semi-solid gel containing an
aqueous buffer. In general, slab has dimensions of a few
centimeter on a side and is capable of separating more than
one samples imultaneously.

11. EXAMPLES OF SLAB ELECTROPHORESIS:
• DNA Gel Electrophoresis:
• In this technique, DNA samples are run through agarose gel with the help of a
potential and differential mobility of different DNA samples is responsible for their
separation
• INSTRUMENTS
• Gel casting trays: composed of U-shaped UV-transparent plastic. The open ends of
the trays are closed with tape while the gel is being cast in between the A and B
walls.
• Combs: These are plastics combs around which molten agarose is poured to
• form sample wells in the gel.
• Electrophoresis buffer: The buffer which is used for the DNA gel
• electrophoresis usually is Tris-acetate-EDTA (TAE) or Tris-borate-EDTA (TBE).
• Loading buffer: components are glycerol and dye.Glycerol helps to allow the
sample to go into the sample wells and dyes help
• visual monitoring of the
• Ethidium bromide: This is used for the detection of the DNA inside the gel.
• Transilluminator: Transilluminator is an ultraviolet light source which is used to
visualize ethidium bromide-stained DNA .

13. ELECTROPHORESIS
• Pulse-Field Gel Electrophoresis(PFGE):
This technique allows investigators to
separate much larger pieces of DNA than
conventional agarose gel electrophoresis. In
conventional gels, the current is applied in a
single direction (from top to bottom).
However, in PFGE, the direction of the
current is altered at regular intervals
• SDS-PAGE Gel Electrophoresis:
In this technique, protein samples are run
through page-gel with the help of a
potentialand differential mobility of
different protein samples is responsible for
their separation according to the mass of
the protein

14. SDS GEL ELECTROPHORESIS
• When the detergent SDS (sodium dodecyl sulphate) is added to
the proteins during the sample preparation, proteins become
negatively charged by their attachment to the SDS anions. When
separated on a polyacrylamide gel, the procedure is abbreviated
as
• SDS-PAGE (for Sodium Dodecyl Sulphate PolyAcrylamide Gel
Electrophoresis).
Two-dimensional SDS-PAGE Gel Electrophoresis
Each amino acid has its own isoelectric point i.e., pI. pI is the pH at
which the mobility of the amino acid is zero. Hence, proteins which
comprise amino acids, do have a characteristic pI value. In two-
dimensional gel electrophoresis, proteins are separated according to
their pI and then they are allowed to mix with SDS.

16. 2) CAPILLARY ELECTROPHORESIS:
• Capillary electrophoresis is a more
sensitive technique and with the
similar principle,
• it could detect samples as small as
few nL.
• high-speed and high resolution.
• The capillary can also be filled
with a gel, which eliminates the
electroosmotic the capillary
allows higher resolution, greater
sensitivity and on-line detection.

17. Types of CGE
• Capillary Gel Electrophoresis(CGE):
• Capillary gel electrophoresis is generally
performed in a porous gel polymer matrix,the
pores of which contain a buffer mixture in
which the separation is carried out.
• used in electrophoresis is a polyacrylamide
polymer formed by polymerizing acrylamide
in the presence of a cross linking agent
)Capillary Zone Electrophoresis:
Capillary Zone Electrophoresis also known as
Free-Solution CapillaryElectrophoresis (FSCE),
simplest ,
used to separate ionic species by their charge and
frictional forces. Separation based on differences
in the charge-to-mass ratio of the analytes

18. • Capillary Isoelectric Focusing (CIEF):
This technique allows amphoteric molecules, such as
proteins, to be separated byelectrophoresis in a pH
gradient generated between the cathode and anode. A
solute will migrate to a point where its net charge is zero.
At the solutes isoelectric point (pI),migration stops and the
sample is focused into a tight zone
• APPLICATIONS OF ELECTROPHORESIS:
• In DNA Sequencing
• In Medical Research
• In Protein research/purification
• In Agricultural testing
• Measurement of hybrid purity
• Identify genetic markers used by plant breeders
• Quality control in processing and other industries,etc