IIT/FIELD MUSEUMHigh School Transformation Project

Biology

*** DO NOT WRITE ON THIS. THIS IS THE CLASS COPY OF

THE LAB. ***
Lab: Using Paper Chromatography to Separate Pigments
BACKGROUND:
The leaves of most plants contain many different pigments. Chlorophyll is the most
abundant and important plant pigment for absorbing sunlight. It exists in two forms:
chlorophyll a (bright green) and chlorophyll b (dull green). Chlorophyll often hides
the other pigments present in leaves. In autumn, chlorophyll levels drop, allowing
other pigments to show their colors. Yellow xanthophyll and orange carotene are
commonly visible at this time. Other pigments may also be present in leaves.
Question: What is the purpose of pigment?

*** PAUSE NOW AND ANSWER THIS QUESTION ON YOUR

LAB REPORT. ***
The individual pigments in a leaf may be separated by the technique of paper
chromatography. Chromatography means, color writing. The separation takes
place by absorption and capillarity. The paper absorbs the mixture of pigments in a
solvent. Capillarity pulls the substances up the paper at different rates depending
upon their specific properties. As a result, the pigments will separate on the paper.
They show up as colored streaks. The pattern of separated pigment colors on the
paper is called a chromatogram.
PURPOSE:
You are a plant specialist working for a natural clothing company. All the garments
manufactured by your company are made from natural plant fibers and are dyed with
natural dyes. One of your companys designers wants you to develop a new yellow
dye. You think one of the plant species growing in the company greenhouse contains
yellow pigment.
You decide to separate the leaf pigments using paper
chromatography.
MATERIALS:
Spinach leaf solution
Chromatography paper strip
100 mL beaker
Solvent- Acetone or isopropyl alcohol, approximately 10 mL
Ruler
Pencil
Scissors
Tape

1|CHR OMAT OGRA PHY

COHORT

123

100709

IIT/FIELD MUSEUMHigh School Transformation Project

Biology

*** DO NOT WRITE ON THIS. THIS IS THE CLASS COPY OF

THE LAB. ***
PROCEDURE:
Part 1
A)
B)
C)

Preparing the Chromatogram.

Draw a line in pencil across the paper 1.5 cm from the bottom.
Starting at the line, taper the bottom of the paper with scissors.
Your chromatogram should resemble Figure A.

Figure A
Chromatogram

Part 2 Adding the pigment.

A) Using the disposable pipette, place 1-2 drops of the spinach solution on
the pencil line.
B) Allow the drop to dry.

Pencil line

Place drops on pencil line

and allow to dry.
Part 3 Running the experiment.
A) Uncover the beaker. DO NOT INHALE THE SOLVENT!
B) Place the filter strip into the beaker so that only the tip soaks in the
solvent.
C) Fold the top of the paper over the edge of the beaker. Be sure the paper
doesn't sink further into the solvent. The green drop CANNOT be in the
solvent.
C) Wait fifteen minutes, or until the pigments fully separate. There should be
different colors of pigment visible on the strip.
E) Remove the filter strip and cover the beaker with plastic wrap.
F) Sketch your pigment bands in your lab report on the outline you made
earlier.
G) Record the colors in TABLE 1.
Part 4 Identifying the pigments.
A) Use the information below to identify which pigments are in your leaf:
Chlorophyll a: bright green-yellow
Chlorophyll b: dull blue-green or khaki
Carotene: orange or gold
Xanthophyll: pale yellow
B) Record your pigment names in TABLE 1.
C) Label the pigment names on your sketch in your lab report.

2|CHR OMAT OGRA PHY

COHORT

123

100709

IIT/FIELD MUSEUMHigh School Transformation Project

Biology
Name ___________________________________________ Date _______________ Period

Lab: Using Paper Chromatography to Separate Pigments

BACKGROUND:
Question: What is the purpose of pigment?

CONCLUSION / ANALYSIS:
TABLE 1
Type of leaf
Spinach

Color of pigment
Orange

Name of pigment
Carotene

Yellow

Xanthophyll

Bright Green

Chlorophyll a

Dull Green / Brown

Chlorophyll b

1. Would your company be able to use this plant as a source for yellow dye?
Explain your answer.

2. Does your data support this statement: Chlorophyll is the most abundant and
important photosynthetic plant pigment? Explain.

3. Why do most leaves contain more than one type of pigment? (HINT: The article on the
next page will help you with this question.)

3|CHR OMAT OGRA PHY

COHORT

123

100709

IIT/FIELD MUSEUMHigh School Transformation Project

Biology
FOR HOMEWORK: After you complete your lab report questions, read the
following article and answer the question at the end of the article.

4|CHR OMAT OGRA PHY

COHORT

123

100709