CHM 256
Prepared By: 1)Iylia Zafirah Binti Amran (2019409608)
2)Nor Fasira Binti Palalloi (2019411488)
3)Nur Nathasya Binti Hendra (2019441618)
Programe Code, Class: AS116, 2A
Programe Name: Diploma in Food Technology
Prepared For: Dr Yanti Yana Binti Halid
Title: Experiment Paper Chromatography
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Table Of Content
Cover page………………………………………………………………... 1
Content…………………………………………………………………….. 2
1.1 Objective ……………………………………………………………… 3
2.0 Introduction …………………………………………………………… 3
3.0 Analysis……………………………………………………………….. 4
3.1 Procedure……………………………………………………… 4
4.0 Results………………………………………………………………… 5
5.0 Discussion……………………………………………………………...5
6.0 Conclusion……………………………………………………………..6
7.0 References……………………………………………………………..6
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1.0 Objective
1. To observe how chromatography can be used to separate mixtures of food colouring
and identify whether certain food colouring are pure substances or mixtures.
2. To identify components of food colouring by calculating Rf values.
3. To obtain a paper chromatogram of various food colouring.
2.0 Introduction
This experiment is one of the simple ways to identify unknown compounds and separate
mixtures. It works based on the compound interaction which is called as separation technique as those
compounds tend to move within a supporting medium. The compounds interact in two phases which
are the mobile and stationary phase. The technique helps in analyzing, identifying, purifying and
quantifying unknown separable mixtures. The mobile phase is either a liquid or gas which moves the
solvent through the stationary phase during the process. The stationary phase is a liquid or solid
component that is fixed in a place for the procedure.
Paper chromatography works majorly on capillary attractions. The capillary attraction which
depends on adhesive and cohesive forces allows the mobile phase to move up the stationary phase due
to created surface tension interaction from the forces. The major types are the paper chromatography,
thin layer, gas chromatography, column chromatography, High performance liquid chromatography.
There are several applications of paper chromatography and other main types of chromatography
techniques. This technique is applicable in Pharmaceutical industries, hospitals, forensic science,
environmental science and manufacturing plants.
This report describes the experiment conducted using paper chromatography to identify an
unknown mixture. This will be done by comparing four known amino acids with the two unknown
mixtures to identify the unknown mixtures. The experiment will also help master the technique and be
able to analyze the movements made by both unknown mixtures and the known amino acids.
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3.0 Analysis
3.1 Procedures
Stationary Phase
1.A piece of chromatography paper was cut into a small piece that fit in a beaker.
2.A baseline was drew using a pensil, roughly 2 centimetres from the base of the paper.
[Link] points was marked where the point will be the point we going to place four food colouring.
[Link] 1 centimetre, first cross was put and evenly place other crosse going along the baseline.
[Link] food colouring was put at the first cross until the last cross which was starting from blue,
yellow, red and green.
6.A capillary tube was used to take in a little bit of the food coloring liquid.
[Link] liquid was placed onto the cross by touching it very lightly and let it spread out on the paper.
8. Step 7 was repeated a few times to get a nice strong colour.
[Link] 7 till 8 was repeated using other food colouring, using a different capillary tube for each colour
so that the colour will not blend with each other.
Mobile Phase
[Link] a small amount of water into a beaker that fit the paper nicely.
[Link] paper was attached onto a glass rod so that the paper can stand inside of the beaker.
[Link] the paper was stand inside the beaker, wait the water levels to rise.
[Link] paper was dried for 15 minutes after the water level has risen within a couple of centimetres of
the paper.
Technique used
Technique used in this experiment is separation technique. The compounds in the mixture separate
themselves based on the differences in their affinity towards stationary and mobile phase solvents
under the capillary action of pores in the paper. Adsorption chromatography between solid and liquid
phases, where in the solid surface of the paper is the stationary phase and the liquid phase is the
mobile phase.
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4.0 Results
Colour Solvent Distance Spot Distance Rf value
(mm) (mm)
Yellow 101 20 0.198
Purple 101 63 0.624
Blue 101 85 0.842
5.0 Discussion
The result that collected was included the solvent distance and spot distance, both in
millimeters (mm). The solvent front is important for calculation of Rf values and spot distance is the
measurement for the applied spots travelling up the chromatogram. By placing the filter paper at the
low-leveled solvent, the colours are separated from the sample. Plus, by using the data in result, the Rf
values for each food colouring can be calculated which is 0.198 for yellow, 0.624 for purple and 0.842
for blue. According to the result in the chromatogram, the food colouring are rise up to stationary
phase and spread its colour, which is unknown colour are similar with yellow colour at first but at the
top it is darker colour that also similar with the blue colour. It means that the unknown food colouring
is a mixtures of yellow and blue colour. Moreover, red is a pure colour because it shows purple colour
at the bottom and orange colour at top.
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6.0 Conclusion
In conclusions, it clearly shows the importance of paper chromatography in helping to
identify unknown amino acids or analyze any other relevant mixtures that has properties of being
separated by the paper. The colors separated the way they did because of the differences in their
molecular characteristics, specifically, their solubility in water and their rate of absorption by the
paper. The most soluble and readily absorbed ink color was the yellow. The least soluble and least
absorbable ink color was the blue.
7.0 References
1. [Link]
2. [Link]
3. [Link]
4. [Link]
[Link]
5.[Link]
periments/General_Chemistry_Labs/Online_Chemistry_Lab_Manual/Chem_9_Experiments/02%3A_
Paper_Chromatography_of_Gel_Ink_Pens_(Experiment)
6. [Link]
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