Download MBBS Paper Chromatography Lecture PPT

CHROMATOGRAPHY

Chromatography is the technique of separation of

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mixtures in to individual component by using

stationary and mobile phase

The word chromatography has been derived from

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greek word `Chroma' meaning color and `graphein'

meaning writing.

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Tswet, Russian botanist is credited for development of

chromatography

PAPER CHROMATOGRAPHY

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Paper chromatography is a partition chromatography.
It is technique to resolve a mixture of substances by

continuous distribution between the stationary and mobile

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phase.

The mobile phase, which is the solvent, has tendency to carry

the substance with it while the stationary phase, which is

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hydrated cellulose fibres have a tendency to retard the
movement of substances relatively as a result of various
interactive forces between it and the substance.
Reagents
1. Solvent

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N-Butanol,Glacial acetic acid and water are mixed in

the ratio of 4:1:5

2. Visualizing Agent

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Ninhydrin solution

3. Standard Amino Acid solution and unknown sample
Types of paper chromatography
1. Descending paper chromatography

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2. Ascending paper chromatography

3. Circular paper chromatography
Procedure

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A circular whatmann number 1 filter paper is taken and from the

centre of circular paper a radius of 1 cm is drawn and

application points are marked.

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A hole is made in the centre portion with the help of compass.

Equal volume of standard solution along with sample is applied

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on the marked spot using capillary tube

Two petri plate of equal circumference are taken so that each

plate fit over one another

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3/4th of one of the petri plate is filled with the solvent

Now circular paper is placed on the petri plate such that wick dips

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within the solvent.

The other petri plate is used to cover, without disturbing the paper.

The run is continued till the solvent reaches the rim of the petridish.

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Solvent front is marked and paper is removed from the solvent and

dried

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Spray the paper with ninhydrin solution and dry it with the help of hair

drier.

Calculate the retardation factor value

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CLINICAL APPLICATIONS

Screening of inborn errors of amino acid metabolism

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like phenylketonuria, maple syrup urine disease.

To analyze different compounds in drugs.

It is also used in testing of antibiotics

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