Chromatography Lab

Tutlalee Wines
April 21, 2023

1 Introduction
Chromatography is the process of separating a mixture into its different compo-
nents through measuring the rate of how quickly the components move through
a medium. This process has applications in medicine, the food industry, and
forensic sciences. Ultimately, it can be used whenever there is a mixture which
needs to be separated into individual components. For example, in ink, multi-
ple components come together to form a mixture. Ink is a homogeneous mix-
ture, which means the components/makeup are uniform throughout the mix-
ture. Components are, by definition, different elements (pure substances) which
have been chemically bonded. Components can be larger or smaller, based on
which elements they are made up of. The larger components move slowly, but
smaller components are rather quick. Using TLC, or thin-layer chromatography,
a mixture is placed onto a thin sheet of chromatography paper which is slightly
submerged into a solvent. Over time, the solvent moves, and moves the mix-
ture along with it. However, the different components of the mixture move at
different rates, which will result in a rainbow-like set of bands in mixtures with
multiple components. Given this information, we can use the chromatography
of a mixture to make assumptions about the mixture’s properties.

2 Materials
-2 beakers
-Water
-Isopropyl Alcohol
-1 washable colored marker (Black)
-1 permanent colored marker (Black)
-4 pencils
-1 ruler
-Calculator
-Safety goggles (if desired)
-4+ chromatography paper strips
-Scissors
-Tape

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3 Safety
Isopropyl alcohol is a hazardous substance. It is drying to skin, gloves can be
worn to mitigate risk. It is also hazardous to the eyes and face. If there is a
risk of splashing, eye protection can mitigate risk. Isopropyl alcohol should not
be directly inhaled.

4 Procedure
1. Fill 1 beaker with isopropyl alcohol, about 1 cm above the bottom of the
beaker.
2. Fill the other beaker with water, about 1 cm above the bottom of the beaker.
3. Prepare 4 chromatography strips, which should be at least 1 centimeter
longer than the depth of the beaker.
4. With pencil, draw a horizontal line 1cm from the end of each chromatography
strip.
5. Draw a dot with permanent marker in the center of the horizontal line on 2
strips.
6. Draw a dot with washable marker in the center of the horizontal line on 2
strips.
7. Tape each paper strip so that it can hangs off of a pencil. The paper strip
should extend down far enough that when the pencil is laid on top of the beaker,
the paper will touch the liquid but the dot will not be submerged.
8. Place 1 permanent marker strip and 1 washable marker strip on top of each
beaker, so that the strip hangs down and touches the liquid.
9. Wait. Let the liquids move no farther up the paper than 1 cm to the top.
10. Measure the solvent front - this is how far the solvent (water or isopropyl
alcohol) moved up the paper, from the 1cm line.
11. Try to discern where each color ends and begins. Measure how far from the
line each color travelled.

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The setup for each beaker should look a bit like this.

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5 Data, Observations, and Pictures

From left to right - washable in water, permanent in water, washable in isopropyl

alcohol, permanent in isopropyl alcohol.
The washable marker separated into a few differently colored components,
but the permanent marker did not have a variety of colors. The washable marker
in water moved very far, about 8.6 centimeters from the line. The permanent
marker in water only moved about 0.5 centimeters from the line. The washable
marker in isopropyl alcohol moved about 3.5 cm. The permanent marker in
isopropyl alcohol moved about 3.5 cm.

Water Isopropyl Alchohol

Permanent marker Moved 0.5 cm, Black/Grey Moved 3.5 cm, Black/Grey
Washable marker Moved 8.6 cm, Blue/Green/Reddish Orange Moved 3.5 cm, Black/Blue/Green/Yellow

6 Analysis
There were considerable differences between the way the different mixtures re-
sponded to different solvents. The permanent marker did not appear to have a
variety of differently-coloured components, which likely indicates that this par-
ticular marker only uses 1 component as a dye, or uses a few elements which
are very similarly colored. The permanent marker moved significantly farther
when the solvent was an alcohol, rather than water. The mixture’s polarity is

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likely somewhat similar to alcohol, and quite different from water. This could
mean that the marker is alcohol based, or it could mean that it is non-soluble in
water, which is a reasonable assumption considering it is a permanent marker.
The washable marker, on the other hand, separated into a variety of compo-
nents which had very distinctive coloring, which likely means that there were
a variety of differently colored dyes used in the mixture. The washable marker
moved much farther in the water than the alcohol, which perhaps indicates that
the marker’s polarity is similar to that of water. This makes sense, considering
the washable marker is meant to be washed.

7 Conclusion
It was quite exciting to watch the procedure, and the process of the alcohol and
water separating the mixtures into their components was very pretty. I made
my best efforts to reduce error by attempting to stick very closely to the pro-
cedure, and ensuring my materials and procedures were consistent throughout
the experiment. However, I think that the varying levels of ink, the inconsistent
depth of the chromatography paper, and the inability to let the experiment last
until the solvent front had reached up to the top of the paper. If I were to do
this experiment again, I think that it would be valuable to come up with a more
secure system to hold the paper strips into the liquid. I would also be more
precise with my measurements. It also might be helpful to let the experiment
last longer. I would ensure that my solvent front was marked, because it was
not easy to see after the papers had dried.
As stated in the analysis, the permanent marker likely only has a few
similarly-colored dyes, or if this is not the case, these components are similar in
size to a degree that there is little visual disparity. The ink is also likely similar
in polarity to alcohol, and very different from water. The washable marker,
however, appears to have vast diversity in its dyes, which at least include some
blue, red, green, and yellow hues. There are quite distinct differences. The
blue compounds are likely smaller, because they travelled faster, and the redder
compounds may be larger, as they travelled slower. The other case involving
washable marker does not have as much of a visual disparity - it was with alcohol,
and if the ink does not have a similar polarity, the results will not be as good.
This also serves the purpose of illustrating the fact that the washable ink must
not align totally in polarity with alcohols. Ultimately, using chromatography,
conclusions surrounding the properties of ink can be effectively deduced.