Jane Doe

Lab Partner-
3.9.2022, CHEM 3106-500
TA-
Experiment 6: Column Chromatography

Purpose:
Students will use column chromatography to separate a product mixture from a Friedel-Crafts acylation of
ferrocene reaction and characterize the product with thin layer chromograph (TLC) and IR Spectrograph,
and other characteristics.
Physical Properties Table:
Reagents & MW g/mol Density Amount mmol Bp (C°) Mp (C°)
Solvents g/ml
Ferrocene 186.04 1.49 - - - 192.6
CH2Cl2 84.93 1.33 3mL .047 39.6 -
Alumina 101.96 - 2-1in - 2980 2000
Methanol 32.04 .791 - - 64.7 -
Hexane 86.18 .659 - - 69 -
acetylferroc 270.10 3.97 - - - 125-127
ene

Safety:

 Dichloromethane (Danger)
o Risk Factors:
 Causes skin irritation
 Causes serious eye irritation
 May cause drowsiness or dizziness
 May cause cancer
 May cause damage to organs through prolonged or repeated exposure
o Safety Factors:
 Obtain special instructions before use
 Do not handle until all safety precautions have been read and understood
 Use personal protective equipment as required
 Wash face, hands and any exposed skin thoroughly after handling
 Wear eye/face protection
 Do not breathe dust/fume/gas/mist/vapors/spray
 Use only outdoors or in a well-ventilated area
 Ferrocene (Danger)
o Risk Factors:
 Flammable solid
 Harmful if swallowed
 o Safety Factors:
 Wash face, hands and any exposed skin thoroughly after handling
 Do not eat, drink or smoke when using this product
 Keep away from heat/sparks/open flames/hot surfaces. - No smoking
 Ground/bond container and receiving equipment
 Use explosion-proof electrical/ventilating/lighting equipment
 Wear protective gloves/protective clothing/eye protection/face protection
 Alumina
o Risk factors:
 Dust may cause mechanical irritation in the eyes
 Dust may cause mechanical irritation in the skin
 Ingestion of large amounts may cause gastrointestinal irritation.
 May cause respiratory tract irritation and lung damage if inhaled
 Chronic inhalation of fine dust may cause lung damage.
o Safety factors:
 Wear appropriate protective eyeglasses or chemical safety goggles
 Wear appropriate gloves to prevent skin exposure.
 Wear appropriate protective clothing to prevent skin exposure.
 Hexane (Danger)
o Risk factors:
 Explosive
 Heating may cause fires/explosions
 Toxic if swallowed
 Fatal in contact with skin
 Harmful if swallowed
 Toxic to aquatic life
o Safety factors:
 May cause eye irritation.
 May cause skin irritation. Toxic in contact with skin.
 May be fatal if swallowed. May cause irritation of the digestive tract.
 Poison by ingestion.
 May cause respiratory tract irritation. May be harmful if inhaled.
 Dichloromethane (Danger)
o Risk factors:
 Causes skin irritation Causes serious eye irritation
 May cause drowsiness or dizziness
 May cause cancer
 May cause damage to organs through prolonged or repeated exposure
o Safety factors:
 Obtain special instructions before use
 Do not handle until all safety precautions have been read and understood
 Use personal protective equipment as required
 Wash face, hands and any exposed skin thoroughly after handling
 Wear eye/face protection
 Do not breathe dust/fume/gas/mist/vapors/spray
 Use only outdoors or in a well-ventilated area
 Diacetylferrocene (Warning)
 oRisk factors:
 Harmful if swallowed
o Safety factors:
 Avoid breathing dust/ fume/ gas/ mist/ vapors/ spray
 Wash skin thoroughly after handling
 Do not eat, drink or smoke when using this product
 Use only outdoors or in a well-ventilated area
 Wear protective gloves/ protective clothing.
 Methanol (Danger)
o Risk factors:
 Harmful if swallowed, in contact with skin or if inhaled.
o Safety factors:
 Avoid breathing dust/ fume/ gas/ mist/ vapors/ spray
 Wash skin thoroughly after handling
 Do not eat, drink or smoke when using this product.
 Use only outdoors or in a well-ventilated area.
 Wear protective gloves/ protective clothing
 If swallowed rinse mouth and call poison control.
 If on skin wash with plenty of water.
 If inhaled remove a person to fresh air and keep comfortable for breathing.
 Wash contaminated clothing before reuse.
 Dispose of contents/ container to an approved waste disposal plant.
 Acetylferrocene (Danger)
o Risk factors:
 May cause eye irritation.
 May cause skin irritation. Toxic in contact with skin.
 May be fatal if swallowed. May cause irritation of the digestive tract. Poison by
ingestion.
 May cause respiratory tract irritation. May be harmful if inhaled.
o Safety factors:
 Wear appropriate protective eyeglasses or chemical safety goggles
 Wear appropriate protective gloves to prevent skin exposure.
 Wear appropriate protective clothing to prevent skin exposure
Procedure:
1. Add 1 mL of CH2Cl2 to one's vial containing one's crude product from Experiment 5. Use this
solution to spot a TLC plate using a spotting capillary.
2. If ferrocene is not provided, prepare a solution by dissolving a tiny bit (a little solid on the tip of a
spatula) in 1 mL of CH2Cl2 and spot the solution on one TLC plate for comparison
3. Develop ones spotted TLC plate with CH2Cl2. Using the ferrocene reference spot, one
determines if there is any remaining ferrocene in one crude product mixture.
4. One will be dry packing the column used in this experiment.
5. Push a piece of cotton to the bottom of the column. It needs to hold the sand and alumina in
place. Add a small amount of sand to the column. Add alumina to the column. A powder funnel
makes this easy. One only need ~2-3 inches of alumina for this experiment
6. Add the solution of the crude product mixture to ~0.3 g of alumina in a small beaker and
evaporate the solvent by gently blowing compressed air over it in the hood.
7. Scrape the product/alumina mixture from the beaker and add it to the top of the column.
8. This is dry loading one's mixture onto the column so that the elution should give nice level bands
as one performs the chromatography experiment.
9. Once the alumina with one's mixture has been added to the column, sand should be added to the
top of the column.
10. Once one adds solvent to the column do not allow the liquid level to drop below the top of the
alumina in the column. If the solvent level is getting low, turn the stop cock to stop the flow while
more of the solvent/solvent mixture is obtained.
11. Add the solvent down the sides of the column. Try to avoid disturbing the solid in the column. A
pipet can be used to add a solvent mixture by putting the tip to the side and slowly adding the
liquid. Once there is a layer (3 or more inches) of liquid on top of the column that can cushion the
addition of more liquid, the solvent mixture can be poured into the column slowly.
12. In this experiment, the compounds one is separating are all colored so one can see when they are
coming off the column.
13. Each band will be collected separately, and the solvent will be evaporated in the hood to isolate
the compound present in the fraction. Collect the colored solutions separately from the colorless
solvent that elutes from the column before and after each band as it moves down the column to
avoid needless evaporation of solvent that does not contain important compound.
14. One can see each band move down the column as it elutes. As it gets near the bottom, and one is
trying to determine if it is coming off the column one can hold a white piece of paper behind the
bottom part of the column to better visualize if the solution is colored.
15. When preparing solvent mixtures for eluting more polar compounds from the column, the
solvents must be mixed to get a homogenous solution.
16. At this point one is ready to start adding solvent. What solvent one use will depend on what one
wants to elute from the column.
17. Does the TLC of one's product mixture show the presence of ferrocene? If so, one will want to
elute this from the mixture first. If not, one will want to proceed to the next solvent mixture to
elute the acetylferrocene in one's mixture.
18. Use hexane to elute the ferrocene from the column. Keep the stop cock open initially as one add
enough hexane to fill the column. This will displace the air that is in the column. Once the liquid
level reaches above the alumina, one does not want to let the solvent level ever be lower than the
alumina while running the column.
19. As one adds hexane one should see a colored band move down the column. Try to collect only
the colored solution in a beaker or flask separately from the colorless hexane that will come
through the column ahead of the band.
20. Once the solution coming from the column is no longer colored the ferrocene has eluted from the
column.
21. Use a 1:1 mixture of hexane: CH2Cl2 to elute the acetylferrocene from the column. If this does
not seem to be move the band efficiently enough use straight CH2Cl2 to elute the column.
22. If one did not have any ferrocene in one's mixture and this is the first solvent mixture to initially
be added to the column, keep the stop cock open initially as one add enough
hexane/dichloromethane mixture to fill the column. This will displace the air that is in the
column. Once the liquid level reaches above the alumina, one do not want to let the solvent level
ever be lower than the alumina while running the column.
23. If one has already eluted ferrocene from the column, start using the 1:1 mixture of
hexane:CH2Cl2 to elute the column.
 24. One should see a band start moving down the column. Try to collect only the colored solution in
a beaker or flask separately from the colorless solvent that will come through the column ahead of
the band.
25. Once the solution coming from the column is no longer colored the acetylferrocene has eluted
from the column.
26. Elution of diacetylferrocene: Most people should have some diacethylferrocene in the crude
product mixture. One should still see colored material on the column at this point if one's mixture
contained this.
27. A 9:1 mixture of CH2Cl2:CH3OH is used to elute the diacetylferrocene band from the column.
28. One should see a band start moving down the column. Try to collect only the colored solution in
a beaker or flask separately from the colorless solvent that will come through the column ahead of
the band.
29. The solvent from each colored fraction is evaporated in the hood using compressed air and gentle
heating.
30. The mass of each isolated solid part of the initial mixture needs to be determined to determine the
actual yields of each product
Data & Observations:

 The starting material:

o .1012g of crude product and it was a dark orange
 The solution went from being a clear liquid to a pale yellow after filtration through the tube.
 The first filtration of ferrocene had no product in the end leading to the inability to perform an IR
Spectrometry
TLC: The method used to view what species are in the mixture before beginning column chromatography
on the mixture and separating the compounds
o On the right side, the orange line represents the ferrocene in the Crude product of the last
solution.
 This line shows that there is ferrocene in the product by its orange color.
 With the line being broken up there were about two different components in the
mixture (maybe three but not enough)
 Movement: N/A
o On the left side of the THC, there is supposed to be a yellow stain, yet it was not
expressed due to not adding enough ferrocene to the spot solution and not enough CHCl2
to the bottom of the beaker
o Products elute in increasing polarity of the solution:
 Acetylferrocene should be on the top of the TLC, diacetylferrocene in the middle
and ferrocene in the bottom, yet it is unclear which compound is in the bottom.
IR Spectrometer: Used to view specific components of each filtration.
Sample 2 Acetylferrocene

 In the IR Spec there is one peak at 1651.51 cm-1 representing a C=C bond, representing a
difluorescence peak
 The broad peak at 3500 cm-1 represents an O-H bond showing how the methyl group did not
dissolve when evaporating the solution
Sample 3 Diacetylferrocene:
  In the sample there are two carbonyl peaks:
o One peak is located at 1651.09 cm-1 shows a C=C bonds, representing a difluorescence
o The second peak at 1660.08 cm-1 shows a C=C bond, represents the monofluorescence
Amount of Product:

Product Name Weight of Container Weight of Container Weight of Product (g)

(g) and Product (g)
Florecene 58.2 58.2022 .0022
Acetylflorecene 58.2036 58.2512 .0467
Diacetyleflorecene 107.228 107.2448 .0168

Results:
Precent Mass of Florecene:
.0022h (mass of ferrocene) / .105g (mass of crude product) x 100 = 2.1%
Precent Mass of Acetylflorecene:
.0467g (mass of acetylferrocene) / .105g (mass of crude product) x 100 = 45.3%
Precent Mass of Diacetylflorecene:
.0168g (mass of diacetylferrocene) / .105g (mass of crude product) x 100 = 16.6.6%
Overall, the reactions did perform correctly into the correct product due to such a low percentage mass
and products showing in the IR Spec.
Discussion / Conclusion:
In this experiment students separated ferrocene, acetylferrocene, and diacetylferrocene from the crude
product by using dry column chromatography. This was done by the TA preparing the column with sand
alumina, and the crude product mixed with alumina. After the product was separated by adding hexane to
elute the ferrocene, a 1:1 hexane:CH2Cl2 mixture to elute acetylferrocene, a 9:1 mixture of
CH2Cl2:CH3OH mixture to elute diacetylferrocene. Each of the products were then evaporated and
loaded to find the IR. Unfortunately, there was nearly no product for the ferrocene elution, merely with
.0022g, so an IR spectrograph was not given for the first elution. The IR spectrograph of acetylferrocene
and diacetylferrocene showed correct products, besides the extra methyl group in the acetylferrocene. The
TLC was not completely comparable due to the ferrocene spot not showing, therefore not providing a
comparison group. This was due to not having enough base solution or ferrocene on the spot. However,
the crude product spot showed and showed roughly two products yet should have been able to compare to
the ferrocene line to find three products. Lastly, the group forgot to take measurements of the lines on the
product TLC spot, so the group was unable to calculate movement of the components. When calculating
mass yield of the products, one could see the initial reactions did perform correctly from the correct crude
product due to the low mass percentage and products showing in the IR Spec.
Post lab Questions:
1. Rank ferrocene, acetylferrocene, and diacetylferrocene in order of increasing polarity. Do the TLC
results from your fractions support this ranking? Explain.
In increasing polarity, ferrocene has the least polarity, then acetylferrocene and lastly diacetylferrocene with
the highest polarity. The TLC unfortunately cannot support this due to the distance traveled by the solvents
being unreadable, yet hypothetically the TLC could be checked with the retention factor having to be equal to
the same order of the distances traveled by the solvents.

2. Rank the solvents used in the experiment in order by increasing polarity.

The solvents would have hexane moth the lowest polarity, due to it being a nonpolar solvent. The 1:1
mixture of hexane and CH2Cl2 and then the 9:1 mixture of 1:1 mixture of hexane and CH2Cl2 being the most
polar solvent.

3. Why do you start with the least polar solvent/solvent mixture and progress to increasing polar
solvent/solvent mixtures when eluting the ferrocene compounds from the column rather than starting with
more polar solvent system and progressing to less polar solvents?
Students will start with the least polar solvent/solvent mixture since the silica gel is very polar. Knowing
this, starting with the most nonpolar solution would cause the polar parts of the mixture to bind strongly to
the silica gel while the non-polar component flows out with the solvent. To ensure elution of the solutions,
students will start with the least polar solvent and progressively increase the polarity as the components are
eluted.

4. How do the thin layer and column chromatography for this experiment compare regarding stationary
and mobile phases?
When comparing TLC and column chromatography regarding stationary and mobile phases, they both have
a stationary phase and CH2Cl2 as a base. The difference between TLC and column chromatography is that
TLC works against gravity and column chromatography works in the direction of gravity. Recognizing this,
the component which travels the least distance on TLC plate will be eluted the last from the column.

5. What key feature in the IR spectrum of acetylferrocene distinguishes it from the spectrum of ferrocene?
A key feature in the IR spectra of the acetylferrocene is that a carbonyl group (C-O) will be absent in the
spectra of ferrocene at about 1700 cm-1. This difference can allow one to distinguish between the IR of acetyl
ferrocene and ferrocene.