Paper chromatography is a technique used to separate mixtures into their individual components. It uses a stationary phase, such as a paper strip, and a mobile phase, such as a liquid solvent, to separate the components. Different components of a mixture travel up the paper at different rates depending on how strongly they interact with the stationary and mobile phases. Paper chromatography has various applications like separating amino acids and carbohydrates and is useful for analysis, identification, purification, and quantification of mixtures.

1. Paper Chromatography
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Physical Inorganic &
Analytical Chemistry
17SE02CH016 Rahul Jinjala
17SE02CH017 Rudra Kansara

2. Content:
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•What is Chromatography?
•Classification Of Chromatography
•Mechanism Of Separation
•Planar Chromatography types
•Paper Chromatography
•Applications of paper Chromatography
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3. Chromatography:
1 Chromatography is a technique for separating
mixtures into their components in order to
analyze, identify, purify, and/or quantify the
mixture or components.
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4. Classification Of
Chromatography:2
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1. Based on mechanism of separation
• Adsorption chromatography
• Partition chromatography
2. Based on phases
I. Solid phase chromatography
• Solid – liquid
• Solid – gas
II. Liquid phase chromatography
• Liquid – liquid
• Liquid – gas

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3. Based on chromatographic bed
I. Planar chromatography
• Paper
• Thin layer
II. Column chromatography
• Packed column
• Open tubular column

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8. “
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• Column Chromatography
The stationary phase is held in a narrow tube
through which the mobile phase is forced under
pressure or by gravity.
• Planar Chromatography
The stationary phase is supported on a flat plate or
the interstices of a paper and the mobile phase
moves through the stationary phase by capillary
action or by gravity.

9. Mechanism Of
Separation:3
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• Partitioning equilibrium (SP : liquid, MP :
liquid/gas)
• Adsorption equilibrium (SP : solid, MP :
liquid)
• Exclusion equilibrium (SP & MP : liquid)
• Ion Exchange equilibrium (SP : solid ion –
exchanger, MP : liquid electrolyte)
• Affinity equilibrium (SP : immobilised ligand,
MP : liquid)

11. Planar
Chromatography
Types:
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• Thin layer Chromatography (TLC)
Separates dried liquid samples with a liquid
solvent (mobile phase) and a glass plate
covered with a thin layer of alumina or silica
gel (stationary phase).
• Paper Chromatography (PC)
Separates dried liquid samples with a liquid
solvent (mobile phase) and a paper strip
(stationary phase).

13. Paper
Chromatography :5
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14. Purpose:
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• Use the technique of paper chromatography to
separate a homogeneous mixture into its
individual components
Uses:
• Separation
• Identification Chromatographic
paper:
• Stationary phaseSolvent:
• Mobile phase

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Capillary action
The movement of liquid within the space of a material due to the
forces of adhesion, cohesion, and surface tension. The liquid is able to
move up the filter paper because its attraction to itself is stronger
than the force of gravity.
Solubility
the degree to which a material (solute) dissolves into a solvent. Solutes dissolve
into solvents that have similar properties (like dissolves like). This allows different
solutes to be separated by different combinations of solvents.
Separation of components depends on both their solubility in the mobile phase
and their differential affinity to the mobile phase and the stationary phase.

16. Experiment:
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• Preparation time : 10 minutes
• Experiment time : 45 minutes
Purpose:
Time required:
• To introduce the principle and terminology of
chromatography.
Costs:
• Less than Rs 50.
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17. Materials:
• Beakers, cover lids
• Distilled water
• Isopropanol
• Filter
• And other stationary materials
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18. Procedure:
1) Preparing isopropanol solutions:
• Prepare 15 ml of the following isopropanol solutions in
approximately labelled beakers:
0%, 20%, 50%, 70%, and 100%
2) Preparing the chromatography strips:
• Cut 6 strips of filter paper.
• Draw a line 1 cm above the bottom edge of the strip with
the pencil.
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19. • Label each strip with its corresponding solution
• Place a spot from each pen on your starting line
3) Developing chromatograms:
• Place the strips in the beaker
• Make sure the solution does not come above your
start line.
• Keep the beakers covered
• Let the strips develop until the ascending solution
front is abut 2 cm from the top of the strip
• Remove the strips and let them dry
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22. Observation:
1) Observe how some of the dyes are made up of more than one
colour
2) Observe how spots of the same colour separated in low
concentrations of isopropanol compared to higher
concentrations
3) Observe when spots of different colours first started separating
in the different concentrations
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0% 20% 50% 70% 100%

24. Application Of
Paper
Chromatography:
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• Separation of amino acids:
Mobile phase: butanol : acetic acid: water(4:1:1)
Detection: spray with ninhydrin reagent
• Separation of carbohydrates:
Mobile phase: ethyl acetate : pyridine water(10:4:3)
Detection: silver nitrate (1 ml in 200 ml of acetone).
40% NaOH in methanol gives brown
spots.

26. References:
1) Physical Inorganic Chemistry
2) https://owlcation.com/stem/What-is-Paper-Chromatography-Principle-Uses-
experiment-video
3) https://www.britannica.com/science/paper-chromatography
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27. Thank You!
Any
questions?
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