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IIT/FIELD MUSEUM—High 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 company’s 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

E) Remove the filter strip and cover the beaker with plastic wrap. Figure A Chromatogram Part 2 – Adding the pigment. There should be different colors of pigment visible on the strip. DO NOT INHALE THE SOLVENT! B) Place the filter strip into the beaker so that only the tip soaks in the solvent. Part 4 – Identifying the pigments. C) Fold the top of the paper over the edge of the beaker. Be sure the paper doesn't sink further into the solvent. A) Uncover the beaker. place 1-2 drops of the spinach solution on the pencil line. F) Sketch your pigment bands in your lab report on the outline you made earlier. Starting at the line. 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. The green drop CANNOT be in the solvent. B) Allow the drop to dry. taper the bottom of the paper with scissors. 2|CHR OMAT OGRA PHY COHORT 123 100709 . G) Record the colors in TABLE 1. THIS IS THE CLASS COPY OF THE LAB. Pencil line Place drops on pencil line and allow to dry. Draw a line in pencil across the paper 1. C) Wait fifteen minutes. C) Label the pigment names on your sketch in your lab report. Part 3 – Running the experiment.5 cm from the bottom.IIT/FIELD MUSEUM—High School Transformation Project Biology *** DO NOT WRITE ON THIS. A) Using the disposable pipette. *** PROCEDURE: Part 1 – A) B) C) Preparing the Chromatogram. Your chromatogram should resemble Figure A. or until the pigments fully separate.

Why do most leaves contain more than one type of pigment? (HINT: The article on the next page will help you with this question.IIT/FIELD MUSEUM—High 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. Does your data support this statement: “Chlorophyll is the most abundant and important photosynthetic plant pigment”? Explain. Would your company be able to use this plant as a source for yellow dye? Explain your answer. 2. 3.) 3|CHR OMAT OGRA PHY COHORT 123 100709 .

IIT/FIELD MUSEUM—High School Transformation Project Biology FOR HOMEWORK: After you complete your lab report questions. 4|CHR OMAT OGRA PHY COHORT 123 100709 . read the following article and answer the question at the end of the article.