How to Perform a Column Chromatography
Column Chromatography is used to separate different chemical components based off of their polarity. This is a
moderately easy way to separate a mixture of compounds into the individual components. It is typically easiest to
separate different colored components, but this method can be used for clear mixtures as well. In this example, we
will have a mixture of ferrocene, acetylferrocene, and diacetyl ferrocene.
You will need:
 Goggles
 Nitrile Gloves
 4 Pasteur Pipettes
 1 Pipette Bulb
 A micro spatula
 6 large tared vials
 Ring Stand and Clamp
 Labels
 Cotton
 A long piece of copper wire
 An air hose with filter head (Pasteur Pipette with some cotton in the end)
 Air supply
 A chemical scale
 Fine Sand
 Alumina powder
 A 50mL Erlenmeyer Flask
 A 10mL graduated cylinder
 Methylene Chloride (Dichloromethane) – (polar solvent)
 n-Hexane – (nonpolar solvent)
 A 60 mg mixture of ferrocene, acetylferrocene, and diacetyl ferrocene
Figure 3 Graduated Cylinder
Figure 4 Air Hose
Figure 1 Graduated Cylinder
Figure 6 Pasteur Pipette
Figure 2 Erlenmeyer flask
Figure 5 Pasteur Pipette Bulbs
Procedure:
Creating sample
1. Obtain the mixture (it should be a dark red color) and put it in the 50mL
Erlenmeyer flask
2. Dissolve the solid in ~10mL of methylene chloride and add a tip of a spatula’s
(pictured right) worth of alumina.
3. Make sure all of the solid has dissolved, if it does not dissolve after about 5
minutes of swirling, add some more methylene chloride.
4. Once all of the solid has dissolved, evaporate the solvent by gently blowing air
on the solution with air hose with filter head. It is very important to insure that
all of the solvent is evaporated or the column chromatography will not
work.
Preparing Column for Chromatography
5. Take a Pasteur Pipette (pictured right) and put a small ball of cotton at the
bottom. You can push the ball of cotton down to the bottom of the pipette by
using the copper wire.
6. Make sure that the cotton is covering the bottom hole, but that it is not tightly
packed. If the cotton is packed too tight, the column might never drain out.
7. Add a small amount of sand on the top of the cotton to make sure that the top is
flat
8. Next, add ~50-60mg of Alumina to the pipette
9. Next, put your sample on top of the Alumina layer
10. Then add a small amount of sand to make sure that the top is flat again
11. Make sure that there is ~3cm of empty space at the top of the pipette so that
you can add the solvents when separating the components
Figure 7 Micro spatula
Sand
Sample
~50-60 mg of
Alumina
Sand
Cotton
Separating the Compounds
12. Weigh 6 labeled vials and record the weights
13. In a 10mL graduated cylinder, add 5mL of hexane and 5mL of methylene
chloride (this will be your 50/50 hexane methylene chloride mixture)
14. Clamp column to the ring stand and put one of the tared vials under the
column to collect the liquid (pictured right)
15. Fill the rest of the column with hexane using a Pasteur Pipette. It is very
important to keep a constant level of solvent on the column or the
column may dry out and crack.
16. As you are adding hexane, you will notice a layer of yellow coming off
from your sample. This is the ferrocene.
17. Collect all of the ferrocene in one vial with hexane. The liquids will drip
slowly (~1 drop every 2 seconds into the vial).
18. Once there is no more yellow in the column, switch the solvent to the
50/50 hexane methylene chloride mixture using a new pipette. You will
start to see an orange layer come from your sample. This is the acetylferrocene.
19. There will be some time between the yellow liquid being collected and the orange liquid coming out.
During this time, excess solvent (which will be clear) will continue to drip out of the column until the
orange layer is at the bottom.
20. Collect all of the clear liquid that drips out of the column into a second vial.
21. Once the orange layer is starting to drip out, change to a third vial and collect all of the orange layer.
22. Once the orange layer is all collected, switch the solvent to 100% methylene chloride using a new
pipette.
23. A red band will start to come off from your sample.
24. Once again, there will be some time between the orange liquid being collected and the red liquid
coming out. During this time, more excess solvent (which will be clear) will continue to drip out of
the column until the red layer is at the bottom.
25. Collect all the clear liquid in a fourth vial until the red layer starts to come off the bottom. This is the
diacetyl ferrocene.
26. Once the red layer starts to come off, switch to a fifth vial to collect all of the diacetyl ferrocene.
27. Once all of the red layer is collected, store the vials in a safe place for about a week with the cap off.
This will allow all of the solvents used to evaporate, leaving only the three separated compounds.