Column Chromatography: Chromatography of a Mixture of Ferrocene and

Acetylferrocene
Purpose:
Column Chromatography is a method used to purify individual compounds from
mixtures of compounds. This procedure was used to separate a 50/50 FerroceneAcetyleferrocne mix into two pure substances. Separation was based on solubility of each
substance with hexane (the solvent and mobile phase) and alumina (adsorbent and
stationary phase).
Procedure:
A microscale chromatographic column is prepared with 4.5 grams of alumina
used as adsorbent and approximately 10 ml of hexane as solvent (slurry packing method).
About .5 mm of sand is placed on top of the adsorbent:

Column is not allowed to run dry as air bubbles could form and this would ruin
the experiment. Next, .090 grams of a 50/50 mix of Ferrocene-Acetyleferrocine is mixed
with about 2.5 ml of hexane and carefully placed into the column through the funnel at
the top. Spout at bottom is opened and hexane is allowed to flow freely down column and
into collecting containers. The mixture separates into two distinct layers that flow down
the column with the hexane:

Yellow liquid is collected in one flask while red liquid is collected right after in
another. Hexane is constantly added to keep proper flow until mixture has been separated
fully and sand is not colored anymore with Ferrocene or Acetyleferrocene.
Results:
After mixture has been separated, one can see that there is a flask with yellow
hexane and a flask with red/orange hexane. The mixture has successfully been separated,
but is still in the solvent, hexane. This is called elution.
Discussion & Conclusion:
The purpose was to separate the two components of the mixture using an
adsorbent (stationary phase) and a mobile phase like the solvent, hexane. Based on each
molecules affinity to the alumina and the hexane they went down the tube at different
rates, thus allowing us to separate them, while the mobile phase carried it down the tube
to be collected in flasks. After separation of the mixture happened one can see a distinct
yellow band and a distinct red/orange band. The yellow portion was lower on the tube
first and this indicates that it was carrying ferrocene, which makes sense because it is the
less polar substance of the two. The acetylferrocene followed as a red band after and this
makes sense because it is the more polar substance of the two due to the existence of a
hydrophilic group attached to it. Since like dissolves like the ferrocene was separated
out first and then the acetylferrocene. Qualitative measurements can be taken by

evaporating the hexane out of the samples and then testing the remaining residues, but
that was not done.
Post Lab Questions:
4. Once the chromatographic column has been prepared, why is it important allow the
level of the liquid in the column to drop to the level of the alumina before applying the
solution of the compound to be separated?
- So the sample added to the column is in the most concentrated solution possible.
5. A chemist started to carry out column chromatography on a Friday afternoon, reached
the point at which the two compounds being separated were about 3/4ths of the way
down the column, and then returned on Monday to find that the compounds came off the
column as a mixture. Speculate the reason for this. The column had not run dry over the
weekend.
- The two compounds could be of very similar affinity to the adsorbent or the
mobile. Also the adsorbent could have been damaged by improper mixing causing effects
similar to air bubbles forming due to column running dry.