METHODS AND MATERIALS

Time and Place of the Study

The experiment was conducted at the PL – 603 Laboratory at San Pedro College, 12
Guzman St. Obrero, Davao City, Davao del Sur. The data was collected on November 22, 2019.

Preparation of the Materials

The materials needed for the duration of the experiment were mortar and pestle, water
bath, 250mL beaker, cellophane, rubber band, filter paper, capillary, aluminum foil, pencil and
leaves. This equipment was requested through the means of San Pedro College Laboratory
Resource Center.

Procedure

1. Preparation of Developer Solution

a. Mix 5mL of ether and 10mL of ethanol in a 250mL beaker.
b. Cover the beaker with the cellophane or aluminum foil.
c. Make sure vapors can’t escape from the beaker.
d. Let it stand for a while to saturate the beaker with the vapors of the solvent.
2. Preparation of Extract
a. Weigh about one gram of leaves sample.
b. Cut the leaves into small sizes and place in mortar and pestle.
c. Add 10mL of acetone while crushing the leaves.
d. When a dark green extract is already achieved, leave it aside.
3. Preparation of Paper and Separation Procedure
a. Using a pencil, mark the filter paper about 1cm from the bottom. Make sure you
don’t mark the paper with your fingerprints.
b. Along this line, spot your sample about 4cm apart. Let it dry.
c. When the spots are dried, immerse the filter slowly into your developer solution.
d. Allow the developer to rise through the paper.
 e. When it is almost on top, remove the paper and immediately mark the line of
solvent.
f. Allow the paper to dry.
4. The Chromatogram
a. When the paper is already dry, there should be three spots staining the paper:
yellow, darker green, and light green. There might be a fourth color which is red,
but often very difficult to separate.
b. Determine the Rf value of each color.
c. Identify each pigment present.