CHM 256 (BASIC ANALYTICAL CHEMISTRY)

EXPERIMENT 14:
PAPER CHROMATOGRAPHY
PREPARED FOR: MADAM NURUL ‘AIN BINTI JAMION
GROUP: AS1142A1
DATE: 16TH NOVEMBER 2022

NAME STUDENT ID

NUR ILLYYIN BT MOHD AMIN 2022777111

FARISHA MADIHA BT SHEIKH FIDZRAL 2022132673

QISTINA NAYLI BINTI AZIZI 2022914381

FIRZANAH ARESYA BINTI AZMI 2022335423

INTRODUCTION

Paper chromatography and TLC are similar in many ways. A strip of paper is used
instead of an adsorbent-coated plate. A thin layer of water on the paper acts as the adsorbent in
place of a solid adsorbent. Paper chromatography is consequently a liquid-liquid partition
method as compared to a liquid-solid method like column chromatography.

Since filter paper is almost completely pure cellulose with minimal contaminants, it may
typically be used for paper chromatography. The majority of the time, filter paper takes moisture
from the air and absorbs it. This absorbed water makes up around 20% of the filter paper's
weight and is typically enough to carry out paper chromatography successfully.

Paper chromatography is the most effective with very polar organic substances because
very polar water molecules form the adsorbent layer. (The solvent is often carried by non-polar
molecules.) The identification of amino acids, which exist as highly polar, dipolar ions species
with complete positive and negative ion charges, is frequently done using paper chromatography.

In this experiment, we'll use paper chromatography to try and separate and identify
various food colours' component elements. The dye used in this experiment is in a concentrated
form, which is typically marketed as a variety of food colours in supermarkets.Rf formula will
be used in this experiment since it represents the relative distance the spot travelled compared to
the distance it could have travelled if it moved with the solvent front.

Rf = 𝐷𝐼𝑆𝑇𝐴𝑁𝐶𝐸
𝐷𝐼𝑆𝑇𝐴𝑁𝐶𝐸 𝑇𝑅𝐴𝑉𝐸𝐿𝐿𝐸𝐷 𝐵𝑌 𝑆𝑂𝐿𝑈𝑇𝐸
𝑇𝑅𝐴𝑉𝐸𝐿𝐿𝐸𝐷 𝐵𝑌 𝑆𝑂𝐿𝑉𝐸𝑁𝑇
OBJECTIVES
To separate food dyes from food colouring using the paper chromatography method.

MATERIALS
● Microcapillary (75mm x 0.5mm ID)
● Filter paper
● 4 colours of food dyes (blue,green,yellow,red)
● Developing solvent ; 0.2%(2g/L) aqueous solution of NaCl ( 50 Ml)
● Plastic wrap
● Stapler

APPARATUS
● 800 mL beaker
● Glass rod

PROCEDURES

1. A line parallel was drawn to the long dimension about 2 cm from both top and bottom
edge of the filter paper.
2. 4 small ‘x’ was drawn on the line beginning about 2 cm from the edge of the paper and 2
cm equal distance from each other.
3. 70 ml of the developing solvent was put in a clean and dry 800 ml beaker and covered
with plastic wrap held in place by a rubber band.
4. The filter paper was laid on a clean table. A tiny spot of each food colour was placed on
the ‘x’ mark by using microcapillary and allowed the spots to dry.
5. Labelled each ‘x’ mark with appropriate letter symbols (for example B, G, R, Y) to
identify the colours.
6. Allowed the spot to dry by gently waving the paper in the air.
7. The filter paper was placed in the beaker containing developing solvent. Make sure the
paper does not touch the wall of the beaker and the spots of the dye must be above the
surface of the solvent in the beaker.
 8. Sealed the beaker immediately with plastic wrap and allowed the solvent to rise up in the
paper.
9. The filter paper was removed from the beaker immediately when the solvent front had
risen up about 2 cm from the top of the paper (until it reached a parallel line drawn at step
1).
10. The solvent front in the paper was marked with a pencil before the solvent evaporated.
11. After the paper has dried, the distance travelled was measured in mm of the spot from the
original line and the distance travelled by solvent front for each of the food colours.
12. The Rf values were calculated and recorded in the table for every coloured spot.

RESULT AND DATA

Food colour Blue Green Red Yellow

Colour of spot Blue Red Yellow Blue Red Red Yellow Red

Distance travelled 106.0 39.0 89.0 106.0 74.0 74.0 85.0 17.0
by spot (mm)

Distance travelled 110.0 110.0 110.0 110.0 110.0 110.0 110.0 110.0
by solvent front
(mm)

Rf value 0.964 0.354 0.809 0.964 0.673 0.673 0.773 0.155

DISCUSSION

The students’ aim was to separate chemicals into different sub-compounds. The method used in
order to separate the coloured dyes was paper chromatography. Paper chromatography is a
method that separates different substances into different compounds in the sample mixture that
eventually travels in distances. The distance travelled by the dyes or chemicals is based on the
intermolecular force of the molecules with the stationary phase as compared to the mobile phase.
The Rf value, also known as retardation factor can be calculated by using the formula of ratio of
the distance travelled by the solute to the distance travelled by the solvent. The formula is shown
as below:

Rf value = Distance travelled by the solute / Distance travelled by solvent front

In this experiment, they have used four different dyes at the beginning. The colour dyes
produced 2 different colours each except for the colour red. Based on Table 1, the distance
travelled by solvent front is 110 mm which is the same for all the four dyes. This will be
considered as the constant variable in the experiment. For Blue coloured dye, two colours were
observed and identified, which were blue and red. The distance travelled by the blue dye was 106
mm whereas the distance travelled by the red colour was 39 mm. By using the formula stated
above, the Rf value of blue dye and red dye formed would be 0.973 and 0.358 respectively. For
Green coloured dye, two coloured spots were also identified which were yellow and blue. Each
had 89 mm and 106 mm distance travelled by the spot respectively. The Rf value for the yellow
spot of green dye would be 0.8 whereas the green spot is 0.964. For red coloured dye, there was
only one colour observed which was red itself. The distance travelled for the red coloured spot is
74 mm. The Rf value of the red spot is 0.673. The fourth coloured dye is Yellow, also produced
two different colours which are yellow and red. The distance travelled by the yellow and red spot
are 85 mm and 17 mm respectively. The Rf values for yellow and red spots are 0.773 and 0.155
respectively.

In this experiment, the students have managed to identify which was in the stationary phase and
mobile phase. The stationary phase is referring to the filter paper used due to the reason that the
filter paper was not in mobile state and stayed put at one place. However, the mobile phase was
when the aqueous solution of sodium chloride, NaCl, pulled the parts of the mixture along with
it. In this case, NaCl would be more suitable to be used as the solvent rather than water. This is
due to the fact that NaCl aqueous solution has a higher polarity and takes shorter time to travel
than water. Depending on the forces that take place, the compound of coloured spots will either
stick with the mobile phase or the stationary phase. Since food dye molecules have different
charges, the highly charged ones would be more polar since it has more ionic bonding as
compared to lower charged ones. This proves that the dyes that have a lower Rf value is due to
the molecules being more attracted to the paper.

Based on the result in the table above, the red spot produced from the yellow food dye has the
Rf value of 0.155, which is the lowest among the coloured spots. This indicates that the red
pigment is less soluble and has a higher polarity than others. The intermolecular forces between
this red pigment and the mobile phase is weak but has a higher bond with the stationary phase.
This caused the absorption to be higher and the pigment travelled the shortest distance compared
to other coloured spots. Meanwhile, the blue spot produced from blue and green dyes resulted as
the furthest spot to travel and has the Rf value of 0.964. This indicates that the blue pigment is
highly soluble and has a lower polarity. Eventually, strong intermolecular forces with the mobile
phase will cause the time taken for it to be absorbed to increase. Hence, the mobile phase will
take the blue pigment further.

Red food dye has only one colour spot, which is red. This is due to it being a primary colour.
Primary colours are the source of all other colours usually used when mixing dyes or paints.
There are supposedly three primary colours which are red, yellow and blue. In this case, only red
food dye was proven to be the primary colour. Blue coloured dye has the composition of blue
and red. On the other hand, green coloured dye produced two different spots which were yellow
and blue. This was expected because green is a secondary colour, which is a result from mixing
two primary colours.

Throughout the experiment, there were several precautions that were taken. Firstly, the parallel
lines marked on the chromatography paper were drawn with a good quality pencil instead of a
mechanical pencil to avoid the disappearance of the mark being dissolved into the solvent. Other
than that, the beaker was always kept sealed at all times during the experiment to avoid the
solvent from evaporating.

QUESTIONS
1. Calculate the Rf value of the spots.
(Calculated in results and data table)
2. Describe the composition of the food colours. Do they consist of single components or a
mixture of dyes?
In blue food colouring, it consists of a mixture of red and blue dye.
In green food colouring, it consists of a mixture of yellow and blue dye.
In red food colouring, it only consists of a single component of red dye.
In yellow food colouring, it consists of a mixture of red and yellow dye.
3. Is there a green dye in the green food colouring?
Yes, there is green dye in the green food colouring.
CONCLUSION

From this experiment, we can conclude that the food colours are all made up of more than one
component of dye, except the red colour food dye. The food dyes were successfully separated by
using the paper chromatography method.

REFERENCES

1. “RF Value - Definition, Calculation, Explanation & Factors affecting RF Values.” BYJU'S,
https://byjus.com/chemistry/rf-value/. Accessed 22 November 2022.

2. Kendall, Emily. “primary colour | optics | Britannica.” Encyclopedia Britannica,

https://www.britannica.com/science/primary-color. Accessed 23 November 2022.

3. Guerrero, Angela. “Calculating retention factors for TLC (video).” Khan Academy,
https://www.khanacademy.org/science/ap-chemistry-beta/x2eef969c74e0d802:intermole
cular-forces-and-properties/x2eef969c74e0d802:separation-of-solutions-and-mixtures-ch
romatography/v/calculating-retention-factors-for-tlc. Accessed 23 November 2022.