Download MBBS (Bachelor of Medicine and Bachelor of Surgery) Latest Paper Chromatography Lecture PPT
CHROMATOGRAPHY
Chromatography is the technique of separation of
mixtures in to individual component by using
stationary and mobile phase
The word chromatography has been derived from
greek word `Chroma' meaning color and `graphein'
meaning writing.
Tswet, Russian botanist is credited for development of
chromatography
PAPER CHROMATOGRAPHY
Paper chromatography is a partition chromatography.
It is technique to resolve a mixture of substances by
continuous distribution between the stationary and mobile
phase.
The mobile phase, which is the solvent, has tendency to carry
the substance with it while the stationary phase, which is
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
N-Butanol,Glacial acetic acid and water are mixed in
the ratio of 4:1:5
2. Visualizing Agent
Ninhydrin solution
3. Standard Amino Acid solution and unknown sample
Types of paper chromatography
1. Descending paper chromatography
2. Ascending paper chromatography
3. Circular paper chromatography
Procedure
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.
A hole is made in the centre portion with the help of compass.
Equal volume of standard solution along with sample is applied
on the marked spot using capillary tube
Two petri plate of equal circumference are taken so that each
plate fit over one another
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
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.
Solvent front is marked and paper is removed from the solvent and
dried
Spray the paper with ninhydrin solution and dry it with the help of hair
drier.
Calculate the retardation factor value
CLINICAL APPLICATIONS
Screening of inborn errors of amino acid metabolism
like phenylketonuria, maple syrup urine disease.
To analyze different compounds in drugs.
It is also used in testing of antibiotics
This post was last modified on 30 November 2021