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