I AM HAFIZ MUHAMMAD WASEEM from mailsi vehari BSc from science college Multan MSC university of education Lahore i love Pakistan and my teachers

1. HAFIZ
MUHAMM
AD
WASEEM
BA BA G
LAHORE

2. Extraction Techniques
(Part. 1)
Lecture 6
UNIVESITY OF EDUCATION
LAHORE PAKISTAN

3. CENTRIFUGATION
• Separates different particles from a
solution
• Separates according to their size, shape,
density, viscosity of the medium and rotor
speed.

4. • The particles are suspended in a liquid medium
and placed in a centrifuge tube.
• The tube is then placed in a rotor and spun at a
define speed.
• Rotation of the rotor about a central axis
generates a centrifugal force upon the particles
in the suspension.

6. FACTORS AFFECTING
CENTRIFUGATION
• Density of both samples and solution
• Temperature/viscosity
• Distance of particles displacement
• Rotation speed

7. CENTRIFUGE MACHINE
• It is a device that separates
particles from a solution
through use of a rotor.
• these particles are usually
cells, subcellular organelles, or
large molecules.

8. TYPES OF
CENTRIFUGATION
• Microcentrifuges
• SMALL BENCHTOP CENTRIFUGES
• GENERAL PURPOSE CENTRIFUGES
• LARGE CAPACITY CENTRIFUGES
• SUPERSPEED CENTRIFUGES
• ULTRACENTRIFUGES
• DENSITY GRADIENT CENTRIFUGATION
• DIFFERENTIAL CENTRIFUGATION

9. Microcentrifuges
• used to process small
volumes of biological
molecules, cells,
or nuclei.
• generally hold 0.5 - 2.0
mL of liquid

10. Continued….
• spun at maximum angular speeds of 12,000–
13,000 rpm.
• small enough to fit on a table-top and
• have rotors that can quickly change speeds.
• They may or may not have
a refrigeration function.

11. SMALL BENCHTOP
CENTRIFUGES
• used for small amount of
material that rapidly sediment
like yeast cells, erythrocytes
etc.,
• they offer maximized capacity
in a compact footprint.

12. Continued….
•Research Applications:
• Cellular Biology,
• Microbiology,
• Genomics / Molecular Biology,
• Proteomics,
• Biochemistry and
• Pharmaceutical Studies.

13. Continued….
•Clinical Applications:
• Clinical Chemistry,
• Clinical Microbiology,
• Hematology,
• Immunology and
• Clinical Studies.

14. GENERAL PURPOSE
CENTRIFUGES
• These centrifuges feature innovative
rotor technologies designed for
improved benchtop performance
and flexibility, greater sample
capacity, and increased speed.
• There is lots of choice of rotors,
buckets and adaptators to fit your
needs.

15. LARGE CAPACITY CENTRIFUGES
• our large capacity centrifuges provide
reproducible separations for
high‐throughput applications such as
blood banking and bioprocessing.

16. SUPERSPEED CENTRIFUGES
• Combine cutting-edge technology,
• high-speed performance, and
• versatile rotor capacities, allow to
maximiz productivity
with impressive acceleration rates.
• .

17. ULTRACENTRIFUGES
• ultracentrifuges and micro-
ultracentrifuges are
designed to deliver
exceptional speed.

18. DENSITY GRADIENT
CENTRIFUGATION
• one of the more efficient methods of separating
suspended particles.
• Density gradient centrifugation can be used both
as a separation technique and as a method of
measuring the densities of particles or molecules
in a mixture.

19. DIFFERENTIAL
CENTRIFUGATION
• selectively spins down components of a mixture by a
series of increasing centrifugation forces.
• This method is commonly used to separate organelles
and membranes found in cells.
• Organelles generally differ from each other in density in
size, making the use of differential centrifugation.
• The organelles can then be identified by testing for
indicators that are unique to the specific organelles.

20. APPLICATION
• Separating chalk powder from water
• Removing fat from milk to produce skimmed milk
• Separating particles from an air-flow using cyclonic separation
• The clarification and stabilization of wine
• Separation of urine components and blood components in
forensic and research laboratories
• Aids in separation of proteins using purification techniques
such as salting out, e.g. ammonium sulfate precipitation

21. Extraction Techniques
(Part. 2)
Lecture 7
UNIVESITY OF EDUCATION
LAHORE PAKISTAN

22. CELL FRACTIONATION
• It allows different parts of a cell to be separated
from each other using centrifugation.
• The purpose is to
isolate/fractionate cell components based on
size and density.

23. • Once the cells have been fractionated,
organelles such as the plasma membrane,
nucleus, and mitochondria can be studied
separately.
• Cell fractionation allows to study the different
parts of a cell in isolation.
• Let's look further into the steps involved in this
process.

24. STEPS INVOLVED IN CELL
FRACTIONATION
• Step One: Breaking the cells or cell lysis
• Step Two: Separation
• Step Three: Collection

25. STEP ONE: BREAKING THE CELLS
OR CELL LYSIS
• Detergents disrupt cell membrane to be opened so
that the contents inside the cell can be obtained.
• Detergents do so because they can interact with
both membranes and parts of the cell that are
soluble in water.

26. • Since detergents can interact with both lipid
(membrane) and soluble (cytoplasmic) parts of
the cell, they allow cellular components to be
mixed or homogenized.
• When the cellular components are mixed, a cell
homogenate, or cell lysate, is formed.

27. • Lysing, or opening the cells, in detergent is usually
combined with a physical method that breaks the cell
further, such as machines that are similar to blenders,
glass beads, or breaking the cell using sound energy.
• Using physical methods in combination with detergents
ensures that all of the cells in the sample get broken, and
you can isolate as much of your cellular fraction as
possible

29. STEP TWO: SEPARATION
• Cell homogenates are separated into fractions by
centrifugation.
• centrifugation produces forces that are thousands of times
higher than gravity, and cellular components are pushed
toward the bottom of the container they are in.

30. Continued….
• The smaller components stay homogenized in the liquid
(labeled supernatant in the image) and the larger
components will move to the bottom.
• Repeating the centrifugation with increasing force allows
smaller cellular components to be separated as seen in
this figure.

32. STEP THREE: COLLECTION
• The way cell fractions are collected will depend on the liquid in
which the cell fractions are centrifuged.
• Cell fractions are usually centrifuged in a medium, or liquid, that
provides osmotic support such as sucrose or Percoll.
• The liquids aid in the separation of cellular components based on
density and size.
• If only one concentration of sucrose or Percoll is used, it is
called differential centrifugation because the different fractions will
be collected by centrifuging the sample several times, as shown in
this figure.

34. FILTRATION
• Filtration is a physical,
biological or chemical
operation that
separates solid matter
and fluid from a
mixture.

35. CONTINUED….
• It requires a filter medium through which only
fluid can pass.
• Solid particles that cannot pass through the filter
medium are described as oversize and
• the fluid that passes through is called the filtrate

36. Hot filtration
• used to separate solids from a hot solution.
• This is done in order to prevent crystal formation in the
filter funnel and other apparatuses
• Therefor apparatus and the solution used are heated
• One of the most important measures to prevent the
formation of crystals is use of stemless filter funnel.

37. COLD FILTRATION
• This method is used when the solid is initially dissolved.
• The solution is cooled down before filtration.
• Cold Filtration method is the use of ice bath in order to
rapidly cool down the solution to be crystallized rather
than leaving it out to cool it down slowly in the room
temperature.
• This technique results to the formation of very small
crystals.

38. DISTILLATION
• A process in which a liquid or
vapour mixture of two or more
substances is separated into
its component fractions of
desired purity, by the
application and removal of
heat.

39. CONTINUED….
• Distillation is based on the fact that the vapour of a
boiling mixture will be richer in the components that
have lower boiling points.
• Therefore, when this vapour is cooled and condensed,
the condensate will contain more volatile components.

40. • At the same time, the original mixture will contain more
of the less volatile material.
• Distillation columns are designed to achieve this
separation efficiently.

41. SOXHLET EXTRACTOR
• invented in 1879 by Franz von Soxhlet.
• designed for the extraction of a lipid from a solid
material.
• Typically, Soxhlet extraction is used when the
desired compound has a limited solubility in a
solvent, and
• the impurity is insoluble in that solvent.

42. DRAWING OF A
SOXHLET
EXTRACTOR
1:Stirrer
2:Stillpot
3:Distillationpath
4:Thimble
5:Solid
6:Siphontop
7:Siphonexit
8:Expansionadapter
9:Condenser
10:Coolingwaterin
11:Coolingwaterout