Introduction

• The word Chromatography is derived from two

Greek words, Chroma means colour and
graphein means to write.
• It is laboratory technique for the seperation of a
mixture.
• Chromatography, literally “color writing”, was
used, and named in the first decade of the 20th
century by Micheal Tswett, primarily for the
seperation of plant pigments such as
chlorophyll, xanthophylls, carotenoids.
 Chromatography Terms
• Chromatograph – Equipment that enables a
sophisticated separation.
• Eluent – Fluid entering column/ solvent that
carries the analyte.
• Eluate – Mobile phase leaving the column.
• Stationary phase/ Immobilised phase –
substance that stays fixed inside the column.
• Mobile phase – Solvent moving through the
column.
• Analyte / Sample –
Mixture whose
individual components
have to be separated
and analysed.
• Solvent – Any
subsatance capable of
solubilizing another
substance.
• Retention time – Time
takes for a particular
analyte to pass through
the system under a set
condition.
Example
 Principle
• Differential affinities of the various components of
the analyte towards stationary and mobile phase
results in the differential separation of the
components.
• Affinity, in turn, is dictated by two properties of the
molecule: Adsorption and Solubility.
• The interplay between the above two factors
determines the differential rates at which the
different components of the analyte will move
through the column.
• Polarity of the compounds possess different affinities
towards the stationary and mobile phase
• .
 Types of Chromatography
1. Based on interactions between sample component and
stationary phase:

Partition Chromatography
• Paper Chromatography
• Thin Layer Chromatography

Adsorption Chromatography
Ion-exchange Chromatography
Gel-filtration Chromatography
Affinity Chromatography
HPLC
2. Based on nature of stationary phase or mobile
phase
Planar Chromatography
• Paper Chromatography
• Thin layer Chromatography

Column Chromatography
• Gas Chromatography
• Liquid Chromatography
 Uses of Chromatography
• Analyze – Examine a mixture, its components,
and their relations to one another.
• Identify – Determine the identity of a mixture or
components based on known components.
• Purify – Separate components in order to isolate
one of interest for further study.
• Quantify – Determine the amount of a mixture
and/or the components present in the sample.
 Real life uses

Hospital
Pharmaceutical company
Law Enforcement
Environmental Agency
Manufacturing Plant