Chromatographic SeparationAlina GunaRyan YoungEdward 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
Separate• Analyze
• Identify
• Purify
• QuantifyComponentsMixture
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.
DataSpot 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
Black Ink
Blue Dye
Red Dye
Orange Dye
SoluteSubstance
DistanceDistance
fR
13434
.
.fR
697.034
3
.fR
3271.0346.1
.fR
Applications Anything involving separating a
solution Medicine
Sports Medicine – Testing for drug usage (Gas chromatography)
Pharmaceutical industries – Check if the right reactants were used.
Criminology & Forensic Science 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