Special Health Sciences High School

City of Dasmariñas, Cavite, Philippines

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Experiment No. 1
Paper Chromatography

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ABSTRACT

Chromatography, which literally means “ color-writing,” is a method of separating the components of a

mixture. This experiments utilizes paper chromatography which is a method that uses filter paper as the
stationary phase. The experiment aims to determine the qualitative analysis of food dyes and to see the purpose
of chromatography which is to separate homogenous mixture or separate mixtures of substances into their
components. During the experiment, four common food dyes was used which was blue, green, red, and yellow.
The food dyes was evaluated under a 0.10% NaCl in water placed in a beaker. Meanwhile, when the group
removed the paper and marked the solvent front, the group saw that the Red, Green, and Yellow produced
another color rather than its base, the Red dye produced Pink, the Green dye produced Yellow and Blue, and
Yellow dye had Red, while the remaining color which is Blue stayed the same. Afterwards, the group calculated
distance traveled by the solute
the Retardation Factor using the formula Rf = . A high Rf value implied
distance travled by the solvent front
that the component used has a high affinity towards the mobile phase, this helped the group expound the
qualitative identification of compounds.
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Introduction

Paper Chromatography is a distinct kind of planar chromatographic method that uses filter paper as the medium
of the experiment. Chromatography means color writing. It is a way of separating a mixture. There are different
types of chromatography such as liquid chromatography (LC), gas chromatography (GC), supercritical fluid
chromatography (SFC), column chromatography, and planar chromatography. Paper chromatography can be
used for qualitative analysis for separation of the mixture. After developing the chromatogram, the spots,
distance of the solvent front and spots can be measured. After collecting all the data needed, RF (retardation
factor) can be determined by dividing the distance traveled by the solute over the distance traveled by the
solvent front. The higher the RF value, the higher affinity towards the medium there is. The objective of this
experiment is to successfully perform a paper chromatography on qualitative analysis on food dyes and
compute for its RF value on different components of the mixtures.

MATERIALS AND METHODS

The materials that were used for the experiment were filter paper, pencil, ruler, beaker, food coloring (blue,
green, red, and yellow), graduated cylinder, and NaCl solution. A line was drawn parallel to the long dimension
of the filter paper about 2 cm from the edge. Four pencil marks were marked on the line drawn in step 1,
beginning 5 cm from the edge of the filter paper and the spacing was about 3cm apart. 70 mL of NaCl water
was measured using a 100-mL graduated cylinder and then transferred to a clean and dry 800-mL beaker. Using
a toothpick, a small spot of food dye was applied on the first dot, same thing went for the rest of the three dots
but with a different color on each dot. The letters B, G, R, and Y were written above each dot to identify the
food dye. The spots were dried after gently waving the paper in the air and afterwards the filter paper was
formed into a cylinder. Then the paper cylinder was placed in the beaker containing the NaCl with the spots at
the bottom. The beaker was sealed immediately with a plastic cover to allow the solvent to rise within the top of
the paper. When the development was finished, the paper was removed from the beaker and the solvent front
was marked with a pencil line.Then the densest part of the band for each color was located and marked by a
pencil. The distance was measured from the point of application to the marked densest part of the band which
corresponds to the distance travelled by each solute. The distance travelled by the solvent front was also
measured from the point of application to the marked solvent front. The Rf values of the colored spots were
calculated.

RESULTS AND DISCUSSION

Results

Paper Chromatography is an experiment to separate the homogeneous mixtures of each colors. In this
experiment, homogeneous mixtures are represented by different food dyes of blue, green, yellow, and red. This
experiment quantifying unknown separable mixtures. By testing the distance travelled by each spot to the
solvent point. It will show that there is a set of variables among all the food dyes. These variables are used in
this experiment 1) Unchanging starting point of the Dyes (20mm or 2cm above the bottom part of the filter
paper) 2) Unchanging ending point of the dyes (10mm or 1cm below the upmost part of the filter paper) after
that, it is needed to expose the filter paper with dyes to the NaCI solution for about 10-15 minutes. The result of
the experimented food dyes will conduct its travelled speed.

Original Component Component Distance Distance RF or

Color of Ink 1 2 traveled by traveled by Retardation
Spot (mm) Solvent Factor
(mm)
Blue Blue 80 mm 82 mm 0.98
Green Yellow Blue 52 mm/ 75 82 mm 0.64 / 0.91
mm
Red Pink Red 10 mm/ 69 82 mm 0.12 / 0.84
mm
Yellow Red Yellow 10 mm/ 59 82 mm 0.12 / 0.72
mm

Discussion

Paper chromatography is usually used to separate the components or color of the mixture. As seen in the table
above, we can see that the blue color did not show any different colors besides its original color. This means
that it doesn’t have any other colors mixed in it. The next is the green color, as we can observe, the green color
separated into 2 colors. The first displayed yellow and second is blue. This proves that when we mix yellow and
blue the result will be green. Another color is red, it showed little color of pink at the beginning and ended with
a red color. This means that red has separated only one color which is pink. The last color is yellow which
showed a separate color that is red on the beginning of the process.

The distance traveled by the solvent NaCl are all the same because the inks are placed in a single sheet of filter
paper and all are dipped at the same time.
The ink spots did not travel at the same rate because every ink has their own solubility. The most soluble and
readily absorbed ink color will travel the farthest while the least soluble would travel the nearest. In this case the
blue color travelled the farthest and yellow color travelled nearest.

The RF or Retardation Factor is the ratio of the distance travelled by the spot to the distance travelled of the
solvent. This factor measures how much the spot retarded compared to the solvent. The blue color showed the
highest retardation factor which means it has a stronger attraction to the solvent (mobile phase). The pink and
yellow color displayed the lowest retardation factor which means it has a stronger attraction to the stationary
phase.

REFERENCES
1. What is the purpose of chromatography in a lab experiment? Retrieved on September 26, 2018
https://www.enotes.com/homework-help/what-purpose-chromatography-lab-experiment-358491