Seperation Techniques – Paper Chromatography

in this video let us look at how we can separate the inks in different colored markers by using chromatography
using a filter paper. we first draw a start line about 1.5 centimeters from the base using a pencil, next we apply
small but concentrated spots of our sample on the piece of chromatography paper. this is important to ensure
that there is enough sample as the individual components will be diluted when they move along the paper. if
the sample spots are too large it will be difficult to distinguish the individual spots on the chromatogram. The
spot is made as small as possible so as to prevent tailing as far as smudging when the inks or dyes are being
separated. chromatography is the technique used to separate small amounts of substances from a mixture
based on the different solubilities of the components in the particular solvent. Now pause the video and think
about what are some of the advantages of using chromatography over the other separation techniques you've
learned. the principal advantage of using chromatography is, that only a very small sample is required for the
analysis. to begin we first add a small amount of a suitable solvent to the beaker, in this case why do you think
water is a suitable solvent ? the choice of solvent depends on the type of sample we are using. in this case as
we are using water-based inks, water is a suitable solvent because the inks are soluble in water. moving on
we now place the chromatography paper and immerse it in the solvent. we also must not forget that we have
to cover the beaker with a lid. why do you think this step is important? covering the beaker is early minimizes
evaporation of the solvent as its travel up the chromatography paper. you can see that as the solvent moves
up the chromatography paper, it carries two different dyes along with it. dyes which are more soluble in the
solvent will migrate faster and further along the chromatography paper. here we see the solvent front moving
upwards. when the southern front has reached near the top we then remove the chromatogram and mark the
solvent front with a pencil line. when do you think this step is important? the tight spots will be traveling up the
chromatogram relative to the measured so in front, knowing the solvent front is important for the calculation of
RF values. notice that also let the solvent front run as high as possible before removing the paper. the further
the solvent front from the start line , we can ensure a better separation for the spot of the dye, for accurate
identification. However we cannot let the solvent front overrun, otherwise we cannot measure the actual
distance covered. let us take a look at the results. we find that many other inks are made of a mixture of dyes,
except for the yellow one, because there is only one spot. we can say that the yellow ink is pure because it is
only made up of one dye. the black ink is a mixture of three different dyes of which the blue dye is the most
soluble in the solvent as it has traveled the furthest. the blue ink is a mixture of two different dyes one
blue and one purple and the purple dye is insoluble in water that's why it remains at the start line and did not
migrate up the paper. for the dyes that were not well separated we can consider changing the solvent. now let
us repeat the experiment but this time let's take a look what will be the outcome if we had drawn the start line
using ink instead of pencil. you can see that the ink being solible where the solvent as well will also move
on the chromatogram and complicate the result. summarizing what we have learned so far. how do you think
you will answer these two questions.
1. Why is the start line drawn in pencil?
A pencil line is made up of graphite and is insoluble in the solvent, hence it will not lead to an
inaccurate chromatogram.
2. Why is the start line not drawn in ink?
A pen/marker is made up of ink and will dissolve in the solvent, thus complicating the results, leading
to an inaccurate chromatogram.

let us repeat the experiment again but this time let's see what happens if the ink spots are below the solvent
level. We observe there are different inks do not get carried up the paper, but instead dissolve in the solvent.
the inks cannot be separated. it is therefore important to ensure that the sample spots are above the solvent
level? this is to prevent a sample from dissolving in the solvent before it's carried out the chromatography
paper. this would prevent the separation of different components of the mixture. now let us summarize the
three different experiment we have done in this video