Paper Chromatography Practical
Aim:
To investigate how paper chromatography can separate different
components in colored ink.
Introduction:
Paper chromatography is a technique used to separate and identify
components in sample mixtures. The procedure involves 2 phases, the
mobile phase which is the solvent, and the stationary phase which is the
filter paper. The separation of the components is caused by the solvent
travelling up the filter paper by capillary action and carrying along the
different components of the ink. The components have different affinities
for the two phases, which is why they travel at varying rates across the
paper. This results in distinct spots and can then be used to calculate its
retention factor (Rf value), which is the ratio of the distance travelled by
the components to the distance travelled by the solvent. The Rf values
are used to identify the components of the mixture by comparing the Rf
values of unknown substances with those of known standards. Paper
chromatography is commonly used for analysing inks, dyes, and
identifying amino acids in proteins.
Variables:
Independent variable Dependent variable Controlled variable
Type of solvent (e.g. Distance travelled Type of
water, ethanol, HCl acid by solvent front chromatography
is used in this practical) Distance travelled paper used
Concentration of the by each Amount of mixture
solvent component applied to the paper
The mixture/substance Rf value The origin
used to separate (e.g. Amount of solvent
ink, amino acid, plant used
pigment)
Methodology:
Materials/Equipment:
Beaker
Grey lid pencil
Filter paper
Binder clip
2 different coloured markers
0.1 mol of Hydrochloric acid (HCl)
Risk Assessment:
Material name Associated risks Safety Environmen
measures taken tal Risk
Hydrochloric Hydrochloric acid in Lab coat HCl is
Acid contact with skin can worn emitted
cause chemical burns Safety by coal
that can be severe. glasses burning,
Hydrochloric acid in the Gloves so air
eyes can cause pollution
blindness.
Coloured Moderate eye irritation Gloves Hazardo
marker Minimal toxic to skin, us waste
contact with mouth or
ingested
Possible skin allergy
Procedure:
1. Pour 200ml of 0.1 mol hydrochloric acid into
the beaker.
2. Draw a line on the filter paper with a grey lid
pencil, 1cm above the bottom of the paper.
This line is called the origin and is made
with a pencil, so it does not interfere with
the experiment.
3. Draw 1 dot each with the coloured marker on
the origin, making sure to press the tip of
the markers strongly and evenly on the
paper.
4. Roll the top of the paper onto a pencil and clip it into place with a
binder clip. It should look like a flag, with filter paper as the flag and
the pencil as the flagpole.
5. Place the pencil with the paper on top of the beaker, with the
bottom of the paper dipped in the solvent (hydrochloric acid).
Ensure that the sample spots on the paper are not below the origin
line, effectively preventing the separation and ruining the
experiment, and that the pencil is longer than the diameter of the
beaker so it can balance stably.
6. Observe the separation and when completed, take the filter paper
out of the beaker and mark the end of the solvent front. Find the
distance travelled by the solvent and the components of the ink.
Results:
A brown marker and a blue marker were used in the experiment, and
these were resulting pigments observed in the chromatogram:
Brown marker = yellow orangish, dark pink and blue (from bottom to
top)
Light blue marker = light blue
The length of the solvent front was 9.0cm. The retardation factor, Rf
value, is the main dependent variable in this practical and is calculated by
dividing the distance travelled by the components by the distance
travelled by the solvent.
The distance travelled by the yellow orangish component from the
brown marker ink was 5cm, the Rf value of the component is 5.0/9.0 =
0.55
The distance travelled by the dark pink component from the brown
marker ink was 6.3cm, the Rf value of the component is 6.3/9.0 = 0.70
The distance travelled by the blue component from the brown marker
ink was 8cm, the Rf value of the component is 8.0/9.0 = 0.88
The distance travelled by the blue component from the light blue
marker ink was 8.7cm, the Rf value of the component is 8.7/9.0 = 0.96
Discussion:
Brown ink marker
This ink was separated into 3 distinct colours, yellow orangish, dark pink
and blue, and indicates that the ink is composed of multiple pigments with
varying affinities for the stationary phase (filter paper) and the mobile
phase (solvent, HCl). The yellow orangish component travelled 5cm and
suggests that the component had an intermediate affinity for the
stationary and mobile phases. The Rf value of the yellow orangish
component, 0.55, means that the component had moderate solubility in
the solvent of HCl. However, in comparison to the Rf values of the other
components, this component had the lowest solubility and travelled the
slowest, having more affinity to the stationary phase.
The dark pink component travelled 6.3cm, a slightly greater distance than
the yellow orangish component, indicating it has a higher affinity for the
solvent compared to the stationary phase, resulting in a higher Rf value of
0.70. This strong affinity to the mobile phase suggests that this
component is more soluble in the HCl solvent than the yellow orangish
component and could possibly be less concentrated in the brown ink
mixture and therefore be ‘lighter’ (have less particles).
The blue component travelled 8.7cm, the furthest out of all the pigments
and had a Rf value of 0.88, the highest out of the 3 Rf values. This
indicates that the blue pigment had the most affinity for the mobile phase
and had high solubility in the solvent of HCl. This could also show that
there is less blue pigment in the brown ink mixture and has the lowest
concentration, the smallest number of particles.
Light blue ink marker
This ink was not separated into different colours, and therefore different
components and travelled the most out of all the components in the
chromatogram. The component did move along with the solvent and
resulted in a straight line of the same pigment, meaning that the ink of
the marker is pure and is not made up of different inks. It is unlikely to
state that the light blue ink was not insoluble in the solvent as none of the
ink remained at the origin. The Rf value of the blue component was 0.96,
this indicates that this component has high solubility in hydrochloric acid
and could possibly be identified with this data.
Advantages & Disadvantages
Pro Con
Simple and requires minimal Complex mixtures cannot be
work separated
Rapid analysis, relatively quick Less quantitative results, unhelpful
separation in statistical analysis
Qualitative (visual) results Effected by environment e.g.
Small sample size required temperature
Intermolecular Forces:
Polar Interactions: The chromatography paper, typically made of
cellulose, is polar and forms hydrogen bonds with polar substances.
Pigments with polar functional groups will have stronger interactions
with the paper and travel more slowly.
This could mean that the yellow orangish pigment from the brown ink
marker could have more polar functional groups than the other two
components.
Non-Polar Interactions: Non-polar pigments interact less with the
polar paper and more with the solvent if the solvent is non-polar. This
will result in these pigments traveling further up the paper.
Hydrogen Bonding: Components capable of hydrogen bonding with
the solvent will be more soluble and travel further, especially when the
solvent is water.
Capillary Action:
The solvent moves up the paper by capillary action, carrying along the
components of the mixture. As the solvent ascends, components that are
more soluble in the solvent and less attracted to the paper move further,
namely having more affinity for the mobile phase. Conversely,
components that less soluble in the solvent and more attracted to the
paper will lag behind, more affinity for the stationary phase.
Conclusion:
In this paper chromatography practical, the aim was to separate and
identify the different components of a coloured ink marker using a solvent.
The experiment successfully resulted in the separation of 2 coloured ink
markers, brown and blue, and the retardation factors were successfully
found. Some possible errors in this practical could be: measuring the
distance travelled by the solvent front and the components (ruler not
accurately placed), observing the pigments in the chromatogram and
identifying the colour (colour-blindness or sight issues can affect this) and
calculating the Rf value (ensuring to measure from the origin to the end of
the pigment, calculator issue). Some improvements for this practical could
be accurately measuring the distances and repeating the practical
multiple times to collect more results and have stronger
analysis/discussions.
Bibliography:
Clark, Jim. “E. Paper Chromatography.” Chemistry LibreTexts, 3 Oct.
2013,
chem.libretexts.org/Bookshelves/Analytical_Chemistry/Supplemental_M
odules_(Analytical_Chemistry)/Instrumentation_and_Analysis/
Chromatography/V._Chromatography/E._Paper_Chromatography.
Mr. Jansen Tan. “Separation Techniques | Paper Chromatography.”
YouTube, 26 Feb. 2017, www.youtube.com/watch?v=uOhefwQBAbI.
“TutorChase.” Tutorchase.com, 2024,
www.tutorchase.com/notes/ib/chemistry-2025-hl/2-2-10-
chromatography-interplay-of-intermolecular-forces.
“What Is the Importance of Intermolecular Forces in Chromatography?”
Quora, 2019, www.quora.com/What-is-the-importance-of-
intermolecular-forces-in-chromatography.
Sidenote: I was not present for this practical and used my friend’s results
