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Paper Chromatography

This document describes an experiment to identify photosynthetic pigments in pechay and papaya leaves using paper chromatography. The experiment involved extracting pigments from the leaves and developing the chromatogram to separate the pigments. Three pigments - chlorophyll, xanthophylls, and carotenes - were identified based on their retention factor (Rf) values, which indicate each pigment's polarity relative to the stationary and mobile phases. Carotenes had the highest Rf value and were most attracted to the solvent, while chlorophyll had the lowest Rf value and greatest attraction to water. The objectives, materials, procedure, results, and conclusions of the experiment are detailed in the document.

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CarlaAbelar
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567 views

Paper Chromatography

This document describes an experiment to identify photosynthetic pigments in pechay and papaya leaves using paper chromatography. The experiment involved extracting pigments from the leaves and developing the chromatogram to separate the pigments. Three pigments - chlorophyll, xanthophylls, and carotenes - were identified based on their retention factor (Rf) values, which indicate each pigment's polarity relative to the stationary and mobile phases. Carotenes had the highest Rf value and were most attracted to the solvent, while chlorophyll had the lowest Rf value and greatest attraction to water. The objectives, materials, procedure, results, and conclusions of the experiment are detailed in the document.

Uploaded by

CarlaAbelar
Copyright
© © All Rights Reserved
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd
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Group 1 Members

Abelar, Maria Louise Carla


Agcaoili, John Alsandair
Andaya, Ana Marie
Antonio, Angela Teresa
Badong, Sophia Veida
Balana, Kevin


Paper Chromatography of Photosynthetic
Pigments
I. Materials
Pechay leaves, papaya leaves, 4 x 8 inches
chromatographic paper, 500ml beaker, 20 ml
developing solvent( Benzene petroleum ether or
acetone- petroleum ether), mortar and pestle,
capillary hematocrit tube, and watch glass.

II. Objectives
The objectives of this experiment are:

1.To identify and know the pigment that is
present in pechay and papaya leaves.
2.To know the fastest and slowest pigment
that moved.
3.To know the relationship of the pigment
to its retention factor value(Rf value).

III. Procedure
Schematic Flow Chart

IV. Results
Papaya Leave Pechay Leaves




Rf value=distance travelled by the pigment from the origin
distance travelled by the solvent from the origin

Data:
Distance travelled by the solvent from the origin = 6.7 cm
For pigments of Papaya and Petchay leaves, respectively:
Green pigment: 1.5 cm; 0.4 cm, Yellow pigment: 3.0 cm;
0.5cm and orange pigment: 6.6 cm; 6.6cm.

Table of Retention Value (Rf value)
Extract

Rf Value
Papaya
Leaves
Petchay
Leaves
Green 0.22 0.06
Yellow 0.45 0.07
Orange 0.99 0.99

V. Explanation
From the results, there is more than one
pigment present in leaves. These are orange pigment of
carotenes, yellow pigment of xanthophylls and green
pigment of chlorophylls. Pigments are substances that
absorb light that is used in photosynthesis. The primary
photosynthetic pigments are chlorophylls but there are
also other photosynthetic pigments such as carotenes and
xanthophylls which are called accessory pigments. These
accessory pigments strongly absorb the wavelengths from
light that are not absorbed by chlorophylls. As the solvent
move up the chromatography plate, the pigments move
along with it. The more nonpolar the pigment, the more
soluble it is in the nonpolar organic solvents, the faster it
will move and the greater distance it will proceed up the
film. The Rf value indicate whether a pigment is more
attracted to the stationary phase (water in the paper) or to
the developing solvent. The greater the Rf value, the more
attracted it is to the developing solvent. This means that
chlorophyll is more attracted to water than xanthophylls
and carotenes because it has the lowest Rf value.

VI. Conclusion
In leaves, there are three pigments present that
absorb light. These are chlorophyll (green), xanthophylls
(yellow) and carotene (orange). The fastest moving
pigment is carotene while chlorophyll is the slowest
because of the polarity of the pigment to the solvent. The
relationship of the pigment to its Rf value is its attraction
to the stationary phase (water) and the developing
solvent. Chlorophyll attained the lowest Rf value and
carotenes have the highest Rf value. This means that
chlorophyll is more attracted to water than the other two
pigments while carotene has the least attraction to water.

References:

Mauseth, J.D.,(2013).Botany: An Introduction to Plant
biology,5
th
edition. Philippines: Mind mover publishing
house, inc. 240 241.

Cobar,M.A.C.S, Laurente,O.S. and
Vasquez,R.D.(2013).Laboratory manual for Pharmaceutical
Botany. Philippines

http://www.ihmc.us/groups/voluntology/wiki/291f0/

1
Obtain a 4 x 8 inches chromatographic paper .
Make a faint pencil line across the paper about an inch
from the tip of the paper(origin) and another faint
pencil line half an inch at the other side of the
paper(solvent front).
2
Macerate leaves of pechay and papaya leaves with
acetone manually with mortar and pestle.
With a capillary hematocrit tube, apply a strip of leaf
extract over the faint pencil mark(origin). Repeat the
application six (6) times then let it dry.
3
Staple both ends of the chromatographic paper,
forming a cylindrical shape. Do not overlap the ends.
Place it in a 500 ml beaker containing 20 ml of
developing solvent. The tip of the paper with the must
be submerged in the solvent.
4
Cover the mouth of the beaker with a watch glass and
do not disturbed set up.
Remove the paper when it reaches the solvent front
and mark the position of the pigments. Compute for Rf
value.

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