Chromatographic Separation
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Transcript Chromatographic Separation
Chromatographic Separation
Alina Guna
Ryan Young
Edward Sun
Jonathan Lam
Mr. Vincent SCH3U3
Contents
Introduction to Chromatography
Adsorption
Paper Chromatography
Experiment
Materials
Procedure
Observations and Calculations
Video
Applications
Conclusion
Sources of Experimental Error
Introduction
Chromatography – Separating
Mixtures
Includes a mobile/stationary phase
Preparative or Analytical
Preparative: Separate to use
components
Analytical: Measure relative proportion
of a substance
5 different types of Chromatography:
Adsorption
Partition
Ion Exchange
Molecular Exclusion
Affinity
Ex: Chromatography in Action
Adsorption Chromatography
When a mobile gas or liquid collects onto a stationary solid
Forms a thin layer of molecules
Solvent travels up adsorbent to meet sample
Continues upwards as a solvent/solute solution (adsorbate)
Different compounds in sample
Different solubilities
Travels different distances
• Analyze
Separate
• Identify
• Purify
Mixture
Components
• Quantify
The other types …
Partition Chromatography
Separation of similar substances by repeated
extraction by two immiscible liquids.
Ion Exchange
Separation of ions and polar molecules
Molecular Exclusion
Particles are separated based on size.
Usually applied to large molecules such as
proteins and industrial polymers
Affinity
Method of separating biochemical mixtures
We focus on Adsorption in our project.
Paper Chromatography
Stationary Phase
Chromatography Paper
Mobile Phase
Unreactive solvent
Primarily used to separate and
identify coloured mixtures
Two-way paper chromatography
Complicated substances
Amino Acids
Solvent’s ability to move
upwards via capillary action
Process of Paper
Chromatography
Experiment
Explain adsorption
What it is
How it is used to separate mixtures
Demonstrate adsorption through paper chromatography
Calculate retention factors of the substances
Safety Concerns
Safety Goggles
Rubbing Alcohol
Spillage/Leakage
Consumption
Ventilation of Area
Permanent Markers
Condition
Wastage
Materials
Glass Jars
Chromatography Paper
Pencils
Rubbing alcohol (70% Isopropyl Alcohol)
Sharpie Pens of various colours
Measuring cup
Ruler
Scissors
Tape
Safety Goggles
Procedure
Use the scissors and ruler to cut equally long
strips of Chromatography Paper. With a pencil,
draw a horizontal line 1 cm above the bottom
edge of each strip.
Using each Sharpie pen, place a spot along the
line of the Chromatography Paper.
Tape each strip onto a different pencil, and
place the pencil onto the jar. Make sure that the
strip of Chromatography Paper is touching the
Isopropyl Alcohol inside.
Measure approximately 15 mL of Isopropyl
Alcohol and pour it in each jar.
Let the strips develop until the ascending
solution is approximately 2 cm from the top of
the paper. Then remove the strips and let them
air dry.
Record your observations and determine the
retention factor.
Data
Spot Colour
Adsorbate
Colour
Distances
Travelled
(Respective)
Distance
Travelled by
Solvent
Retention
Factor
(Respective)
Black
Blue, Orange,
Red
4.3cm, 1.6cm,
3cm
4.3cm
1, 0.3271,
0.6977
Gray
Purple
4.3cm
4.3cm
1
Blue
Blue
3.9cm
3.9cm
1
Green
Green, Yellow
4.1cm, 4.4cm
4.4cm
0.9318, 1
Light Green
Green, Yellow
4cm, 4.4cm
4.4cm
0.9090, 1
Orange
Orange, Red
3.1cm, 4cm
4cm
0.7750, 1
Burgundy
Burgundy
4cm
4cm
1
Green (2)
Green, Yellow
3.8cm, 4.2cm
4.2cm
0.9048, 1
Orange (2)
Orange, Red
3.2cm, 4.2cm
4.2cm
0.7619, 1
Burgundy (2)
Burgundy
4cm
4cm
1
Calculations
DistanceSubstance
Rf
DistanceSolute
Black Ink
Blue Dye
Red Dye
Orange Dye
4.3
R
1
f
4.3
3
R
0.697
f
4.3
1.6
R
0.3271
f
4.3
Applications
Anything involving separating a
solution
Medicine
Criminology & Forensic Science
Sports Medicine – Testing for drug
usage (Gas chromatography)
Pharmaceutical industries – Check if
the right reactants were used.
DNA sequencing (Capillary
electrophoresis)
Analyzing evidence (Thin-layer
chromatography)
Petrochemical Industries
Checking the purity of fuel (Gas
chromatography)
Sources of Experimental Error
Chromatography Paper
Condition
Measurements of Distance
Marker Dot Size
Experiment Modifications
More Solutes
Different Colours
Different Rf Values
E.g. Food Colouring,
Homemade mixture
Different Solvents
Different Concentrations of
Isopropyl Alcohol
Experiment Modifications
Testing known Solvents
E.g. Known chemical formula
Easier to find relationships
Record speed of Solution
Relationships with respect to
time
Conclusion
Intermolecular Forces
Stronger forces = Slower speed
traveling upwards
Longer time in stationary phase
Retention Factor
Can help identify an unknown
compound
Surface tension of Markers
All solutes tested were polar