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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

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Chromatographic Separation

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

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

  3. 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 • 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 • 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 • 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

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

  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 • 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

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