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Chromatography
Very useful technique in organic chemistry based on differential adsorption.
Used to separate components in a mixture (solid or liquid).
Thin layer chromatography (TLC) is also used to monitor the progress of a reaction.
Chromatography
What do we need to perform a chromatographic separation?
Adsorbent: Silica gel (silicon dioxide), also called stationary phase.
Eluent: solvent used to move your compound trough the silica gel, also called the mobile phase. Your compound mixture to be separated. Patience and chemical intuition.
Chromatography
More polar molecules stick to the adsorbent longer. Less polar molecule separate more easily from the adsorbent. When this happens, separation occurs.
SiOH SiOH
SiOH
SiOH SiOH SiOH SiOH SiOH SiOH SiOH SiOH SiOH
Stationary phase
To be separated
Chromatography
More polar solvent move the molecules more efficiently Less polar move the molecules less efficiently Separation occurs
Least polar Alkanes Toluene Diethyl ether Chloroform Acetone Ethyl acetate Ethanol Methanol (CH3OH)
Most polar
Column Chromatography
Load the silica gel plus eluent into the columnthis is called column packing
Column Chromatography
Using a Pasteur pipette, load your compound that was dissolved in a minimum of solvent onto the silica. Your test solution will then add the eluent. Do not let your column run dry!!
Use different capillary for each solution. make solution of approx. 1-2 mg of sample in 1 ml of solvent. Spot 2-3 times
Try to make small spots
Dont let the solvent front run off The top of the plate!!
What to do today?
Using diethyl ether, you will extract a mixture of three compounds from paprika ( Capsanthin, Capsorbin and b-carotene). The three compounds have different polarities, thus can be separated using chromatography. You will spot the mixture on a TLC plate, develop using 15% Et2O and 85% heptane. Calculate the Rf for each spot. Record data in your notebook with the color of the spots. Using the procedure in your handout you will perform a column chromatography, this time you will increase the polarity of the solvent (eluent) gradually. Asses with TLC