Name: ______________________________________ Date:__________________ Grade: ________

TITLE: “Is the topic of the lab session.”

Separation of photosynthetic pigments by chromatograph [0]

TYPE OF LABORATORY

Descriptive With hypothesis [1]

OBJECTIVE: “Given by the teacher will, defines the goal of the lab session.”

Use the separation of mixture technique of chromatography to compare the number of

pigments in two taxonomically different plants [0]

VARIABLES: Dep: ______________________ Indep:__________________________

Control: _______________________________________________________[3]

RESEARCH QUESTION: “Is a question that includes the two variables and is a cause-effect
question stated like How does _____ affect _______? Where the variables are written in the spaces”

How does the type of plant affect the type of pigments in the leaf? [0]

BACKGROUND: “Definitions and main ideas about the topic of the lab session that allow you to
understand what is going on in the laboratory” It may be homework to do previous the lab.
Leaves contain many chemical compounds, such as pigments used in photosynthesis, for example, the
commonly and (inaccurately) known as chlorophyll is actually a combination of five different
pigments (or family of pigments), beta carotene, chlorophyll a, and chlorophyll b, etc.

Chromatography separates pure substances from a mixture, such as a cell extract. This technique can
be used to separate the photosynthetic pigments of a leaf so that they can be identified.
Chromatography is based on different substances having different solubilities. Solvents are usually
mixtures of water with organic liquids like ethanol or propanone (alcohols). The extract of plant
material is dissolved in a solvent, which is used to separate the different pigments through a medium
such as filter paper. The relative distances traveled by different pigments depend on the type of
solvent used. [0]

HYPOTHESIS: “a sentence describing what you think will happen and why you think it will happen.
Must include the variables* you are going to measure and how you think they are going to behave.”

__________________________________________________________________________
__________________________________________________________________________
_______________________________________________________________________[ ]

* Variable is the characteristic you are going to measure the can be dependen and Independent
METHODOLOGY: “Will describe what are you going to do, usually step by step.”

1. Cut a filter paper strip to ensure it fits in the gas jar

2. Rule a pencil line across not along the strip of paper 30mm from one end.
3. Follow steps 1 to 6 in Figure 1. to create the plant concentrate for plant
FIGURE 1. HOW TO CREATE THE PLANT CONCENTRATE

4. Using a dropper, place a drop of the pigment solution at the center of the pencil line. This is the
origin of chromatogram
5. Allow the pigment spot to dry. you can use a fan to speed up the process
6. Place a second drop on the first. Repeat as many as 20 times so that you build up a small but
concentrated spot of pigment.
7. Repeat steps 4 to 6 of figure 1 for the plant B sample. Label the spots with a pencil. Figure 2.
shows a summary of these steps.
FIGURE 2. HOW TO STAGE THE CHROMATOGRAM

NOTE: In figure 2, there

is only one spot in the
paper, your paper needs
to have two spots one
of each plant

8. Using a ruler to
measure distance, pour
the solvent into the gas
jar to a depth of
approximately 15 to 20
mm. Place a lid over the
gas jar so that the
atmosphere inside remind
saturated with vapor.
9. Place JUST the bottom edge of the chromatography paper (one close to the pigments) into the
solvent so that it is just touching. Make sure the pigment spots are not immersed the
chromatography sheet should be placed so that the origin is just above the level of solvent. See
Figure 3.
FIGURE 3. HOW TO PLACE THE CHROMATOGRAM PAPER

10. The solvent front will rise up the paper by capillary action and
separate the pigments. Any solutes dissolved in the solvent will be
partitioned between the organic solvent (the moving phase) and
the water, which is held by the surface of the paper (stationary
phase). The more soluble a solute is in the solvent the further up
the paper it will move.

11. Wait until the solvent is close to the top of the paper, remove the strip of paper, and let it dry.
12. Record your results in the Results section. [0]

MATERIALS: “What you will bring to the lab.”

1. Lab coat 2. Fresh Leaves of two plants. Green grass, spinach, or hibiscus [1]
2. Aluminum foil 3. Pencil, ruler, and goggles 4. Cotton
REAGENTS: “The substances and material we will use in the lab and are consumables”
1.
2.
3.
LABORATORY EQUIPMENT: “material from the lab that is not a chemical substance.”
1. Centrifuge 2. Two test tubes 3. Mortar 4. Hot plate
2. Gas Jar 5. Beaker 6. Measure cylinder 7. Glass rod 8. Dropper

WARNING
● Always follow the safety labels of the liquids or solutions
● Alcohol is highly flammable and must be handled with care
● Do not expose matches or bunsen burner in the vicinity of the experiment
● Use eye protection or goggles
● Centrifuge tubes must be balanced prior to centrifugation

RESULTS: “In this space, you will record what you observe in tables, diagrams, and graphs. The
tables must include the title, heading, and units when possible. The diagrams include a title and
description and the graphs include labels of the axis, proper scale, units of the variables in the
labels, and a title”
1. Draw a diagram of the chromatogram resulting
2.
List the Plant A Pigments List the Plant B Pigments

[6]

ANALYSIS: “in this section, you must contrast your hypothesis with your results and describe your
findings”
__________________________________________________________________________
__________________________________________________________________________
__________________________________________________________________________
__________________________________________________________________________
__________________________________________________________________________
__________________________________________________________________________
__________________________________________________________________________
_______________________________________________________________________[6]

CONCLUSION: “Accept or reject the hypothesis you wrote initially and explain why you accept or
reject the hypothesis considering your results. Short Sentence

_________________________________________________________________________

_______________________________________________________________________[2]

REFERENCES: “quote the book or website or the notes of your notebook that you use in the lab.”
1. Andrew Davis, Internal Assessment for Biology, Skills for success. Hodder Education.
2. 2.9 Separation of Photosynthetic Pigments by Chromatography (Practical 4) [0]

CHECK LIST PREPARATION OF THE LAB

Read all the parts of the lab guide and complete, the objective, hypothesis, type of
laboratory, and materials.
All the girls in the group understand the objective, hypothesis, methodology and
expected results
All girls in the group know what materials to bring to the practice
All the girls in the group recognize the elements
All girls in the group understand the safety measures necessary for the lab [0]