TECHNICAL PAPER

LAB REPORT: PAPER CHROMATOGRAPHY

JOHN MICHAEL E. LUNAR

12 - STEM HONESTY

Lucban Academy
Senior High School
Lucban,Quezon
2020-202
 Abstract

Chromatography is a technique for separation of the components of a mixture on the basis of relative amount
of each solute distributed between a moving fluid stream, called the mobile phase, and a stationary phase.
Chromatography is an essential biophysical technique that allows the separation, identification, and purification of
the components of a mixture for qualitative and quantitative analyses. Proteins can be purified based on features
such as size and shape, total charge, hydrophobic groups present on the surface, and binding capacity with the
stationary phase. Four separation methods based on molecular traits and interaction type use mechanisms of ion
exchange, surface adsorption, partition, and size exclusion. Other chromatographic processes are based on the
stationary bed, including column, thin-layer, and paper chromatography. Column chromatography is one of the
most familiar methods of protein purification.

Introduction

The reason of this experiment is to observe how chromatography can

be utilized to isolated blends of chemical substances. Chromatography serves primarily as
a apparatus for the examination and partition of blends of chemical substances. Chromatography
is employing a stream of dissolvable or gas to cause the components of a blend emigrate in an
unexpected way from a narrow starting point in a particular medium, within the case of
this explore, channel paper. It is utilized for the refinement and confinement of different substances. A
chromatographically unadulterated substance is the result of the division. Since refinement of
substances is required to decide their properties, chromatography is an irreplaceable instrument within
the sciences concerned with chemical substances and their responses (Friedman, 2017).
Chromatography is additionally utilized to compare and portray chemical substances. The
chromatographic arrangement of sorbed substances is related to their nuclear and atomic structures.
A alter in a chemical substance delivered by a chemical
or organic response frequently changes the solvency and movement rate. With
this information, changes or changes can be recognized within the substance (Friedman, 2017).
In all chromatographic separations, there's an important relationship between the dissolvable, the
chromatography paper, and the blend. For a specific blend, the solvent and the paper must be chosen so
the solvency is reversible and be particular for the components of the mixture. The most prerequisite, in
spite of the fact that, of the solvent is to break up the blend requiring to be isolated.
The permeable paper utilized must moreover assimilate the components of the blends specifically and
reversibly. For the division of a mixture, the substances making up the mixture must
be equitably scattered in a arrangement, a vapor, or a gas. Once all of the over criteria have been met,
chromatography can be a basic instrument for isolating and comparing chemical mixtures (Friedman,
2017).

Background

Chromatography is used to separate mixtures of substances into their components. All forms of
chromatography work on the same principle.
They all have a stationary phase (a solid, or a liquid supported on a solid) and a mobile phase (a
liquid or a gas). The mobile phase flows through the stationary phase and carries the components of the
mixture with it. Different components travel at different rates. We'll look at the reasons for this further
down the page.
In paper chromatography, the stationary phase is a very uniform absorbent paper. The mobile
phase is a suitable liquid solvent or mixture of solvents.

Objective

1. To perform simple Separation Methods for common mixtures.

2. To use paper chromatography to determine the Retention Factor of common Black
Ballpen inks.
3. To compare the Retention Factor, Rf, of common Black Ballpen Inks.
Methodology

The materials used for this lab are:

Paper (Bond Paper, Oslo Paper, Filter Paper)
Black Ballpen (Any brand)
Solvent (Water, Rubbing Alcohol, Vinegar)
Clothespin (sipit)
Small Plate or Dish

Procedure:
1. Cut-out a 2” x 5” rectangle for each paper type: Bond Paper, Oslo Paper, Filter Paper.
2. Mark the starting line for your ink 1 cm from the edge and a finish line 10 cm after the
starting line.
3. Place three dots of ink at least 1 cm away from each other along the starting line.
4. Prepare one of your solvent in a small plate or dish.
5. Clip the paper with the starting line hanging at the end above the solvent dish. Once the
solvent has reached the finish line, remove the paper from the solvent and start
measuring.
6. Measure out the length of the traveled distance of the ink from the starting line to the end
of its traveled distance. Record your data in the sample table below.
7. Repeat the same steps for the other solvents and paper types.
8. Calculate for their Retention Factor, Rf

. Use the formula:

Rf=distance l2igment migrated (mm) distance solvent front migrated

Explains the measurements and working equation (if any)

You've conceivably utilized paper chromatography as one of the primary things you've ever

learned in chemistry to confine blends of dark ink for illustration, dark ink that make up a particular
ink. This can be a simple illustration to require, so let's begin from there.
Assume you've got three blue pens and you need to figure out which one of them
was utilized to compose a letter. Tests of each ink are seen on a pencil line drawn on a sheet of
chromatography paper. Any of the ink within the report is broken up within
the least conceivable amount of an successful dissolvable, which is additionally contained on the same
line. The pens are numbered 1, 2 and 3 and the message is seen within the graph.

The paper is suspended in a jostle with a shallow coating with an suitable dissolvable or a

combination with solvents in it. It is fundamental that the sum of the dissolvable is underneath that of
the spots on it. The following chart does not show specifics of how the report is suspended since there
are so numerous potential ways to do so, since the graph is cluttered. In some cases the paper
is truly collapsed into a free envelope and secured to the beat and foot with paper clips.
The barrel is fair standing within the foot of the jostle.
 The reason for covering the holder is to form beyond any doubt that the climate within
the container is immersed with dissolvable vapor. Saturating the environment within the measuring
utencil with vapor stops the dissolvable from vanishing because it rises up the paper.
 As the dissolvable gradually voyages up the paper, the distinctive components of the
ink blends travelat diverse rates and the blends are isolated into diverse colored spots.
 The chart appears what the plate might see like after the dissolvable has moved nearly to the top.

It is decently simple to see from the ultimate chromatogram that the write that composed the

message contained the same dyes as write 2. You'll too see that write 1 contains a mixture of
two diverse dark colors - one of which may well be the same as the single color in pen 3.
It is very unlikely that the two confusing spots will have the same Rf values in the second solvent
as well as the first, and so the spots will move by a different amount.
The next diagram shows what might happen to the various spots on the original chromatogram.
The position of the second solvent front is also marked.

You wouldn't, of course, see these spots in both their original and final positions - they have
moved! The final chromatogram would look like this:

Two way chromatography has completely separated out the mixture into four distinct spots.
 If you want to identify the spots in the mixture, you obviously can't do it with comparison
substances on the same chromatogram as we looked at earlier with the pens or amino acids examples.
You would end up with a meaningless mess of spots.
You can, though, work out the Rf values for each of the spots in both solvents, and then compare
these with values that you have measured for known compounds under exactly the same conditions.

Result and Disscusion

Result

Paper Type Solvent Type Ballpen Brand Distance Retention

Traveled by the Factor, Rf
Ink (cm)
Bond Paper Water FlexStick 5 cm, 5 cm, 5cm 0.5, 0.5, 0.5
Bond Paper Alcohol FlexStick 10cm,10cm, 8 1, 1, 0.8
cm
Bond Paper Vinegar FlexStick 3cm, 2 cm, 1 cm 0.3, 0.2, 0.1

VINEGAR ALCOHOL WATER

Discussion

In water, the distance travel of ink in the first dot is 5 cm and its retention factor is 0.5. In the second
dot, its distance travel is5 cm and its retention factor is 0.5. The last dot his distance travel is 5cm and
the retention factor is 0.5.
In the alcohol, the distance travel of the first dot is 10 cm and its retention factor is 1. In the second
dot its distance travel is 10 cm and its retention factor is 1. The last dot is his distance travel is 8 cm
and the retention factor is 0.8.
In the vinegar, the first dot his distance travel is 3 cm and its retention factor is 0.3cm. In the second
dot, its retention factor is 2 cm and its retention factor is 0.2. The last dot is its distance travel is 1 cm
and its retention factor is 0.1.

There are different comes about within the relationship of solvents to dark ink that

I utilized but utilizing dissolvable water and alcohol these two solvents are to some degree similar to
each other within the result. In case we see that they are almost the same within
the remove travel within the to begin with speck but the moment and third speck the remove travel
is very near.

The cause and effect of this experiment utilizing three solvents and one dark ink we saw

the separate travel of each dab that they were distinctive separations when the ink come
to utilizing three solvents water, alcohol and vinegar. I moreover found out
its retention factor since I utilized the equation.
Conclusion

This experiment was geared for students to have fun with paper chromatography and to
determine which colors are really in the different solvents that were used.
Initially chromatographic techniques were used to separate substances based on their color as
was the case with herbal pigments. With time its application area was extended considerably.
Nowadays, chromatography is accepted as an extremely sensitive, and effective separation method.
Column chromatography is one of the useful separation, and determination methods. Column
chromatography is a protein purification method realized especially based on one of the characteristic
features of proteins.

REFERENCES

Singh,P. (December, 2019) Chromatography (Paper and Thin-layer Chromatography): Techniques and
Applications. Retrieved from
https://www.researchgate.net/publication/338107778_Chromatography_Paper_and_Thin-lay
er_Chrom atography_Techniques_and_Applications
Friedman,J. (2017, April 21). Paper Chromatography Report. Retrieved form
https://www.biologyjunction.com/paper_chromatography_report.htm
Clark,J. (2016, July).Paper Chromatography. Retrieved form
https://www.chemguide.co.uk/analysis/chromatography/paper.html