1. Chromatographic Separation
Alina Guna
Ryan Young
Edward Sun
Jonathan Lam
Mr. Vincent SCH3U3

2. Contents Introduction to Chromatography Applications Conclusion
Adsorption
Paper Chromatography
Experiment
Materials
Procedure
Observations and Calculations
Video
Applications
Conclusion
Sources of Experimental Error

3. Ex: Chromatography in Action
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

4. 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
Identify
Purify
Quantify
Separate
Mixture
Components

5. The other types … Partition Chromatography Ion Exchange
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.

6. Paper Chromatography Stationary Phase Mobile 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

7. Process of Paper Chromatography

8. Experiment Explain adsorption
What it is
How it is used to separate mixtures
Demonstrate adsorption through paper chromatography
Calculate retention factors of the substances

9. Safety Concerns Safety Goggles Rubbing Alcohol Ventilation of Area
Spillage/Leakage
Consumption
Ventilation of Area
Permanent Markers
Condition
Wastage

10. Materials Glass Jars Chromatography Paper Pencils
Rubbing alcohol (70% Isopropyl Alcohol)
Sharpie Pens of various colours
Measuring cup
Ruler
Scissors
Tape
Safety Goggles

11. 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.

12. 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, ,
Gray
Purple 1
Blue
3.9cm
Green
Green, Yellow
4.1cm, 4.4cm
4.4cm
0.9318, 1
Light Green
4cm, 4.4cm
0.9090, 1
Orange
Orange, Red
3.1cm, 4cm
4cm
0.7750, 1
Burgundy
Green (2)
3.8cm, 4.2cm
4.2cm
0.9048, 1
Orange (2)
3.2cm, 4.2cm
0.7619, 1
Burgundy (2)

13. Calculations
Black Ink
Blue Dye
Red Dye
Orange Dye

14. 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)

15. Sources of Experimental Error
Chromatography Paper Condition
Measurements of Distance
Marker Dot Size

16. Experiment Modifications
More Solutes
Different Colours
Different Rf Values
E.g. Food Colouring, Homemade mixture
Different Solvents
Different Concentrations of Isopropyl Alcohol

17. Experiment Modifications
Testing known Solvents
E.g. Known chemical formula
Easier to find relationships
Record speed of Solution
Relationships with respect to time

18. Conclusion Intermolecular Forces Retention Factor
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