CHROMATOGRAPHY

*David, Maria Ariane Agatha Victoria B. , *Santos, Nanette D., Violago, Mary Abelene , Rojales,
Franchesca Nicole
SCHOOL OF CHE-CHM-BE-MSE
MAPUA UNIVERSITY
Muralla St. Manila City, Philippines

ARTICLE INFO ABSTRACT

VOLUME NO: 2 Paper Chromatography is used in separating

colored chemicals, it is a method in which the
ISSUE NO. 5 purity of compounds is tested and identified. In
chromatography, the dyes are observed to have
DATE SUBMITTED moved, the distance the dyes have moved
during the process is typically less than the
DATE ACCEPTED distance from the mobile phase Based on the
results, it can be deduced that the Rf values are
affected by polarity, that ishow the compounds
adsorbs the solvent. The concept, like dissolves
like can be used In this context, it is known that
ethanol is a polar compound and shall therefore
interact with polar compounds. , it can be said
that Methylene Blue, Indigo Carmine are polar
dyes since they have low Rf values, compared
to methyl orange, who has a greater Rf value
The values proved to us that using paper
chromatography is one of the most efficient and
fast ways in determining the presence of a
certain molecule/analyte

KEYWORDS: paper chromatography, retention

factors, polarity

*David, Maria Ariane Agatha Victoria B. CHM145L/T; Mapua University ; School of CHE-CHM
*Santos, Nanette D. Mapua University ; School of CHE-CHM

INTRODUCTION
Paper Chromatography
Chromatography was developed by a Russian
scientist named Mikhail Tsvett, he was looking
for a method to separate the unseen red and
yellow pigments from normal green leaves, so
what he did was he crushed these leaves and
put them into a solution and found that when this
solution was mixed with a certain powder, a
number of color would appear in different areas
of the powder. Paper Chromatography is used in
separating colored chemicals, it is a method in
which the purity of compounds is tested and
identified. Paper chromatography is a popular
choice because it is a quick and relatively an
inexpensive procedure. One major advantage of
paper chromatography is the simplicity of the
equipment(s) required in order to do it, namely:
a suitable type of paper that will act as the
stationary phase, the sample(s) themselves, a
beaker and a solvent, an alcohol is usually used
in this type. A typical paper chromatography is
illustrated in Figure 1.
Retention Factor (Rf)
In chromatography, the dyes are observed to
have moved, the distance the dyes have moved
during the process is typically less than the
distance from the mobile phase. It is possible ot
measure the difference in distance by
calculating the retention factor value.[1]
*David, Maria Ariane Agatha Victoria B. CHM145L/T; Mapua University ; School of CHE-CHM
*Santos, Nanette D. Mapua University ; School of CHE-CHM
Figure 1. A set-up of Ascending Paper
Chromatography
I. EXPERIMENTAL SECTION
Apparatus Used
600 mL Beaker, Capillary Tube, Petri dish,
Chromatogram Paper
Chemical Used
Methyl orange, Methylene Blue, Ethanol,
Fluorescein, Erythrosine, Indigo Carmine,
Unknown
An 8x22 cm chromatogram paper was obtained
from the laboratory assistant, then a light pencil
was drawn parallel to the longer side of the
chromatogram paper 2cm from it’s edge, then,
ten equally spaced dots which are 2 cm apart
were drawn along the vertical edge along the
line of the paper. Then using a capillary tube,
the chromatogram was spotted with dyes in the
order of I, E+I, E, then MB, F+MB, F, then MO,
MO+MB, MB. Contrary to the instruction in the
manual, the students interchanged the positions
of the MO and I dyes in the chromatogram
paper, then the paper was rolled by connecting
the two shorter edges using a staple wire, after
obtaining a cylindrical shaped paper, the rolled
chromatogram was placed inside a beaker
containing 5ml of 5% ethanol, the chromatogram
paper was placed inside the beaker with the
spots at the bottom and it was ensured that the
sides of the beaker was not touched, then the
beaker was covered with a petri dish, then the
chromatogram paper was allowed to develop for
about 15 minutes or until the solvent has
reached about 2 cm from the upper edge of the
paper, then the chromatogram was unrolled, and
the solvent front was traced immediately with a
pencil, then the chromatogram was then air-
dried. The distance traveled by the solvent was
measures and also the distance traveled by the
dyes, then the Rf values was determine and
recorded. [2]
RESULTS AND DISCUSSION
Table 1. Rf values in centimeters
Figure 2. Developed Chromatogram
Based on the results, it can be deduced that the
Rf values are affected by polarity, that ishow the
compounds adsorbs the solvent. The concept,
like dissolves like can be used In this context, it
is known that ethanol is a polar compound and
shall therefore interact with polar compounds.
Therefore, it can be said that there are polar
dyes and will absorb the ethanol easily and
*David, Maria Ariane Agatha Victoria B. CHM145L/T; Mapua University ; School of CHE-CHM
*Santos, Nanette D. Mapua University ; School of CHE-CHM
have lower retention factors than the non-polar
dyes. Because the non-polar dyes will have little
to no interaction with the polar solvent, ethanol.
From the Figure above, it can be said that
Methylene Blue, Indigo Carmine are polar dyes
since they have low Rf values, compared to
methyl orange, who has a greater Rf value. In
handling the chromatogram, it is advisable to
handle the edges in order to prevent the paper
from getting wet with oils or other contaminants.
The Rf values were computed with the following
formula:
Rf= Distance Substance relative to solvent front
Length of solvent front
CONCLUSION
From the results above, it can be said that the
students were able to separate organic
compounds using chromatography, compute Rf
values and identify unknown compounds using
Rf values. The values proved to us that using
paper chromatography is one of the most
efficient and fast ways in determining the
presence of a certain molecule/analyte. Any
errors made could be contributed by that air was
able to pass through any small opening on the
beaker on the waiting time.
REFERENCES
[1] https://sciencing.com/steam-distillation-vs-simple-
distillation-8407975.html
[2]De Castro, K. Baluyut, J. G , Organic Chemistry
Laboratory for Chemists Part 1
 REFERENCES

[1] https://sciencing.com/steam-distillation-vs-simple-
distillation-8407975.html

Retrieved 24 October 2017

*David, Maria Ariane Agatha Victoria B. CHM145L/T; Mapua University ; School of CHE-CHM
*Santos, Nanette D. Mapua University ; School of CHE-CHM