PAPER CHROMATOGRAPHY

J.V. DE GUZMAN
DEPARTMENT OF CHEMICAL ENGINEERING, COLLEGE OF ENGINEERING
UNIVERSITY OF THE PHILIPPINES, DILIMAN QUEZON CITY, PHILIPPINES
DATE PERFORMED: FEBRUARY 11, 2015
INSTRUCTORS NAME: ALLAN KENNETH REGUNTON

ANSWERS TO QUESTIONS
1. Why is the chromatogram developed in an essentially closed
system?
The chromatogram is kept in an essentially closed system in
order to prevent it from acquiring contaminants and other substances
that can hinder in the separation of the organic pigments. It also
prevents the developing solvent used from evaporating since most
solvents used in paper chromatography are highly volatile, and some
can be flammable and toxic.
2. What are the considerations in choosing a chromatographic
solvent?
The chromatographic solvent holds large significance in
separating the components of the sample. If the solvent is polar, the
spots will all at the top of the plate only and if nonpolar, the spots will
still be at their initially positions. In both cases, R f will be zero. As
such, polarity must be taken into consideration when choosing the
appropriate solvent. Usually, the chromatographic solvent is a mixture
of polar and nonpolar solvents combined in appropriate ratio. The
properties of the compound to be separated must also be taken into
account since it will help identify the appropriate ratio of the polar and
nonpolar components of the chromatographic solvent to be used. The
nature of the solid support is also of important consideration because it
will help in providing the appropriate concentration of solvent without
degrading the material.
3. Compare and contrast between normal phase and reverse
phase chromatography.
Normal phase and reverse phase chromatography both take
advantage of the polarity of the constituents of the sample and make
use of a stationary and mobile phase to separate them from each
other. In normal phase chromatography, the stationary phase is the
polar part while the mobile phase is less or nonpolar. However, in
reverse phase chromatography, the reverse is true: the stationary
phase is the less or nonpolar while the mobile phase is polar.
4. Explain how iodine crystals help to visualize the other
pigments.
Iodine crystals will form iodine vapor which in turn will form a

yellow or brown complex or stain with almost all organic compounds

except saturated alkanes. When spots formed from the developing
solvent faded away, iodine vapor forms iodine stains with these
pigments that can be easily observe. Iodine is reversible and is
nondestructive which makes it a convenient visualization technique. It
is also useful in detecting carbohydrates since it forms black spots with
iodine.
5. Suggest one method that can be used to visualize highly
conjugated compounds but are not colored.
Ultra-violet light detection is also a commonly practiced method
that helps in visualizing compounds that are not very noticeable using
ordinary solvents. It is a nondestructive technique that will visualize
any compounds that absorb UV radiation. It makes use of phosphor as
the absorbent that fluoresce in UV light. A compound, even if not
colored, is detected if dark or black spots are observed on the plates
since the compound blocks the glow or fluorescence of the absorbent.
If the compound fluoresces too, it will appear as a bright spot on the
plate when illuminated with UV light.
6. Given a mixture that contains the following compounds: 2chloropentane, pentanoic acid, cyclopentane and pentanal.
Arrange the following compounds in increasing R f if the
mixture is ran using paper chromatography with the same
solvent used in the experiment.
The polarity ranking of the functional groups in increasing order is:
amide>carboxylic
acid>alcohol>ketone~aldehyde>amine>ester>alkyl
halides> ether>alkane. As such, if the given compounds are arranged in
increasing polarity, it will become: pentanoic acid>pentanal>2chloropentane>cyclopentane. Note that the paper chromatography uses
petroleum ether mixed with acetone at a ratio of 9:1 as the developing
solvent. Since the developing solvent is mostly composts of the nonpolar
petroleum ether than the polar acetone, it is expected that it will dissolve
well the less polar compounds. As such, as the solvent ascends in the
chromatographic paper, it will carry first the nonpolar compounds and their
distance travelled together with Rf value will be greater than the more polar
ones. Therefore, in order of increasing Rf: pentanoic acid<pentanal<2chloropentane<cyclopentane
REFERENCES:
Adronov, R. Stains for Developing TLC Plates. McMaster University
Department of Chemistry and Chemical Biology. [Online]. 2015.
http://www.chemistry.mcmaster.ca/adronov/resources/Stains_for_Devel
oping_TLC_Plates.pdf (accessed February 15, 2015).
Columbia University. Experiment 8: Chromatography. Columbia University
Intensive
Seminars
in
Modern
Chemistry.
[Online].
2007.
http://www.columbia.edu/cu/chemistry/ugrad/hssp/EXP_8.html
(accessed February 15, 2015).
Ophardt, C.E. Organic Functional Group Polarity and Electrostatic Potential.

Virtual
ChemBook:
Elmhurst
College.
[Online].
2003.
http://www.elmhurst.edu/~chm/vchembook/213organicfcgp.html
(accessed February 15, 2015).
University of Toronto Scarborough. Thin Layer Chromatography Theory.
Chemistry
Online
UTSC.
[Online].
2014.
http://webapps.utsc.utoronto.ca/chemistryonline/tlc.html
(accessed
February 15, 2015).