Paper Chromatography

of Sugars
Magdurulan, Tricia Andrea N.
C308
Describe the specific mobile and
stationary phases involved in the
chromatographic system used.
Paper Chromatography
A type of partition (separation) chromatography which uses paper
(Whattman paper no. 1 (no.1 relates to pore size)) as the porous medium
where solutes are separated
components of a solution are separated as a result of differential
affinity to the;
stationary phase
water trapped in the interstitial spaces of the paper (NOT the paper itself)
fibers of the paper has cellulose which binds with water via H-bonds
polar liquid
mobile phase
less polar liquid (as compared to pure water in the stationary phase)
 Differential affinity of the sample molecules to mobile and stationary phases result to
partition.

Since the stationary phase is polar, the sample molecules which are also more polar
- are attracted to it
- has greater affinity to it
- are soluble in it
- are adsorbed to it hence they travel less distance

Since the mobile phase is less polar, the sample molecules which are also less polar
- are attracted to it
- has greater affinity to it
- are soluble in it
- are taken along as the mobile phase travels hence they cover more distance
 Mobile phase
can be solid or liquid
organic developing solvent (Butanol:Ethanol:Water)

Stationary phase
can be solid or a liquid
Fibers of cellulose on chromatography paper (Whatman filter paper)
What is 2-dimensional paper
chromatography and discuss how this
technique is done.

What is the advantage of 2 dimensional

chromatography using one solvent
system?
Two-Dimensional Paper Chromatography
It has a more complex set-up which is used to separate
complex mixtures.
This allows a high degree of separation

Steps:
1. Solvent is placed at the bottom of the chamber and the
filter paper saturated with the stationary phase is then
kept inside the chamber.
2. After few hours the filter paper is turned 900 clockwise
and the tank is filled with a different type of solvent.
3. Again the filter paper is turn 900 clockwise and used
another solvent.
What is the principle behind the
visualization of color of the spots
on the chromatogram?

Give the chemical reaction involved.

 The production of colored spots on the
chromatogram is a result of dehydration of the
sugars producing FURFURAL DERIVATIVES.

PINK COLORED
Aniline acid COMPLEX: 5-carbon
oxalate sugars

Aniline acid BRONW COLORED

oxalate COMPLEX: 6-carbon
sugars
In case the solvent ran off the paper how
will you compute for the Rf values?
Ratio of Fronts (Rf)
Rf = distance traveled by solute or unknown
distance traveled by solvent front

extent of migration of the solutes, it is dependent on

nature of the solvent (polarity)
quality of the paper used (pore size)
distance travelled by solvent and solute fronts
not affected by temperature
If the solvent runs off the paper

Rf = distance traveled by solute or unknown

distance traveled by the sugar that migrated farthest
Based on the Rf values obtained, which
sugar is most likely your unknown?