https://drive.google.com/open?id=0B7ewR1jVZ4tsVDc3dmctenBFdGs A presentation slide on various types of chromatography. You Need to download to view the side notes of some slides .

1. Chromatography
Md. Hadiull Islam SUST, BD

2. Chroma means – color.
Graphein – to write.
Chromatography.

3. A little history
 Color writing
 New forms of chromatography

4. Principles
 Partition of the solute between two phases/solvents.
 Mobile phase and a Stationary phase.
 The mobile phase.
 The stationary phase is a porous solid
matrix.
 Interaction.

5. Classification
 Partition
 Adsorption
 Ion-exchange
 Gel-filtration
 Affinity
 High performance liquid
chromatography
Based on interactions between sample component and
stationary phase:
Base on nature of stationary phase or
mobile phase:
 Planar- It may be Paper or Thin layer
 Column- it may be Gas or Liquid

6. Partition
Chromatography
Partition Chromatography:
 Paper Chromatography.
 Thin Layer Chromatography.

7. Paper
Chromatography
 An analytical technique for the separation and identifying
mixtures that are either coloured or can be made coloured
 It is a liquid partition chromatography
 Used for separation of amino acids, sugars, sugar
derivatives & peptides
 The stationary phase- Paper (Filter paper)
 The mobile phase -Mixture of immiscible
solvents

8. Procedure
 At first a line is drawn on the filter paper.
 Colors are placed that line.
 Placed in suitable solvent like ethanol or water.
 Wait.
 Mark the last places of different color.
 Calculate the distance.
 Calculate Rf.
Rf = Distance travelled by color/Distance
travelled by solvent
Rf <= 1.

10. Thin Layer
Chromatography
 Similar like Paper Chromatography.
 Used to identify colorless substances.
 Silica gel is used .
 Use of UV ray.

12. Adsorption
chromatography
 The adsorbents such as silica gel, charcoal powder and calcium
hydroxyapatite are packed in to a column in a glass tube– Stationary
phase.
 The sample mixture in a solvent is loaded on this column.
 The elution is carried out by a buffer system (mobile phase).
 The most weakly held fraction moves fastest.

14. Ion–Exchange
Chromatography
Procedure:
 The stationary phase is bound with and anion and another anion is
bound with it also (For C.E.C) Like
(X+A-) + B+
Stationary phase
 The desired cation in mobile phase is passed through a column E.g. C+
 After passing the new form is (X+A-) +C+ . And B is washed with
elution .
 Then a huge high concentration of another cation(D+) is added and C+
is exchanged with D+.
 Cation Exchange Chromatography(CEC)
 Anion Exchange Chromatography (AEC)
 Cation Exchange Chromatography(CEC)
For anion exchange the reverse is
done.

16. Gel filtration
chromatography
or Molecular Sieve
chromatography
Procedure:
 A desired gel bed is prepared with substances like agarose or
Sephadex G 50 and mixed with buffer like tris buffer
solution.
 Then it is taken in a column and wait until buffer is left.
 A porous network of specific size is built within the gel bed.
 It will trap specific sized protein molecules .
 Then mixtures of proteins are poured with buffer carefully
 The small sized proteins are trapped and remaining larger are
passed out.
 Then those desired small sized desired proteins are also
collected in more purified form .

18. Affinity
chromatography
Procedure:
 At first the stationary phase is complexed with some specific type of molecules
for example for specific antibody specific antigen .
 Then mixtures with desired compound is passed . E.g. human blood
serum with specific antibody.
 Those antibody will bind with those pre fixed specific antigen and
remains will eluted away .
 Then by increasing pH or salt concentration those antibody
are collected by elution.

20. High performance
Liquid
Chromatography
 Completely like adsorption chromatography.
 The difference is here a pump is used to generate pressure for substance
those do not pass faster if that is mobile phase.
 And a computer based detection system is there.

21. Gas liquid
Chromatography
Procedure :
 A inert gas (He , N ) works as mobile phase .
 Samples specially organic volatile substances are injected through a
hot injector.
 Samples get volatile and interact with a stationary phase which is
placed after injector in a column depending on their polarity and
volatility.

22. Conclusion
From here have learned various types of
chromatography
.
In medical, pharmaceutical ,chemistry biology
the impact of chromatography can not be
finished with words .