PHILIPPINE COLLEGE OF SCIENCE AND

TECHNOLOGY
Old Nalsian Road, Nalsian, Calasiao, Pangasinan, Philippines 2418
Tel. No. (075)522-8032/Fax No. (075)523-0894/Website: www.philcst.edu.ph
ISO 9001:2015 CERTIFIED, Member: Philippine Association of Colleges and
Universities (PACU),

NAME: _________________________________ DATE: ______RATING _________

PROGRAM, YR. & BLK. ___________ INSTRUCTOR: MRS. Virginia Arenas Lumague

ACTIVITY NO. 5
PAPER CHROMATOGRAPHY
I. THEORY:
Paper chromatography is an analytical method used to separate colored
chemicals or substances. It is primarily used as a teaching tool, having been
replaced by other chromatography methods, such as thin-layer chromatography
The principle involved can be partition chromatography or adsorption
chromatography. Partition chromatography because the substances are
partitioned or distributed between liquid phases. The two phases are water held
in pores of the filter paper and the other phase is a mobile phase which passes
through the paper. When the mobile phase moves, the separation of the
mixture takes place. 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, wherein the solid surface of the paper is the
stationary phase and the liquid phase is the mobile phase.

Paper Chromatography Diagram

 PHILIPPINE COLLEGE OF SCIENCE AND
TECHNOLOGY
Old Nalsian Road, Nalsian, Calasiao, Pangasinan, Philippines 2418
Tel. No. (075)522-8032/Fax No. (075)523-0894/Website: www.philcst.edu.ph
ISO 9001:2015 CERTIFIED, Member: Philippine Association of Colleges and
Universities (PACU),

II. OBJECTIVES:
Differentiate among the chromatographic techniques
Perform successfully paper chromatography
III. MATERIALS:
Filter paper Micro pipettes isopropyl alcohol
Pencil Beaker Funnel
III. PROCEDURE:
1. Obtain a pre-cut chromatography paper from Laboratory Technician.
2. Sketch a light pencil line about 1cm. from and parallel to, the edge of the
shorter side.
3. Create three equidistant light pencil dot marks along this line.
4. Using separate micropipettes, make small sports of yellow, green and blue food
colors on each dot mark. Handle the paper at the edges only. Allow the spots to
dry.
5. Roll the paper (spots marks outside) to form a cylinder and staple the edges
without letting them touch.
 PHILIPPINE COLLEGE OF SCIENCE AND
TECHNOLOGY
Old Nalsian Road, Nalsian, Calasiao, Pangasinan, Philippines 2418
Tel. No. (075)522-8032/Fax No. (075)523-0894/Website: www.philcst.edu.ph
ISO 9001:2015 CERTIFIED, Member: Philippine Association of Colleges and
Universities (PACU),

6. Dilute 10 ml. of isopropyl alcohol with 5 ml. of water in a small beaker and
transfer this solution slowly to the bottom of a clean 400 ml. beaker by pouring it
through a long stemmed funnel to avoid splashing and wetting the sides of the
beaker.
7. Put the rolled chromatography paper carefully into the center of the beaker,
cover the mouth of the beaker tightly with aluminum foil and do not move the
beaker anymore from its place.
8. Allow the chromatography to develop for about 1 hour or until the solvent has
reached about 1 cm. from the top of the paper.
9. Take out the paper, mark the height of the solvent rise, and allow it to dry.
10. Detach the stapled edges and encircle each colored spot with a pencil.
11. Mark the center of each circle with the pencil line and measure the distance
traveled by each color. Measure also the distance traveled by the solvent.
12. Calculate the Rf value of each colored spot.

Rf = distance traveled by the spot

Distance traveled by the solvent
13. Submit the chromatography together with your report.
Food color applied Color of Spots Distance traveled Rf value
obtained by each color
RED 2 Brown- 1.5 cm
Pink – 3.5 cm
BLACK 2 Black- 2.0 cm
Violet- 2.5 cm
BLUE 2 Blue violet- 3.5 cm
Yellow- 1.0 cm
 PHILIPPINE COLLEGE OF SCIENCE AND
TECHNOLOGY
Old Nalsian Road, Nalsian, Calasiao, Pangasinan, Philippines 2418
Tel. No. (075)522-8032/Fax No. (075)523-0894/Website: www.philcst.edu.ph
ISO 9001:2015 CERTIFIED, Member: Philippine Association of Colleges and
Universities (PACU),

DISTANCE TRAVELLED BY THE SOLVENT ____6.0 cm.________

RED

BLUE

BLACK
 PHILIPPINE COLLEGE OF SCIENCE AND
TECHNOLOGY
Old Nalsian Road, Nalsian, Calasiao, Pangasinan, Philippines 2418
Tel. No. (075)522-8032/Fax No. (075)523-0894/Website: www.philcst.edu.ph
ISO 9001:2015 CERTIFIED, Member: Philippine Association of Colleges and
Universities (PACU),

COMPUTATION FOR CALCULATING Rf value

RED

BLUE

BLACK

IV. CONCLUSION:
1. What is the significant of Rf values?
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2. What is the basis for the separation of components of mixtures by paper
chromatography?
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 PHILIPPINE COLLEGE OF SCIENCE AND
TECHNOLOGY
Old Nalsian Road, Nalsian, Calasiao, Pangasinan, Philippines 2418
Tel. No. (075)522-8032/Fax No. (075)523-0894/Website: www.philcst.edu.ph
ISO 9001:2015 CERTIFIED, Member: Philippine Association of Colleges and
Universities (PACU),

___________________________________________________________________
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3. Give 3 important applications of chromatography.
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ASSESSMENT: The laboratory Activity will be assessed with a maximum score of

60 points. The rubric below will guide you on how your answer will be rated.
 PHILIPPINE COLLEGE OF SCIENCE AND
TECHNOLOGY
Old Nalsian Road, Nalsian, Calasiao, Pangasinan, Philippines 2418
Tel. No. (075)522-8032/Fax No. (075)523-0894/Website: www.philcst.edu.ph
ISO 9001:2015 CERTIFIED, Member: Philippine Association of Colleges and
Universities (PACU),