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1.

Floating Leaf Disk Photosynthesis Lab


Introduction:
Photosynthesis is a process that converts carbon dioxide into sugars such as glucose using energy from the sun. When light is
absorbed by pigments in a leaf, the energy absorbed is used to incorporate the carbon dioxide into organic molecules in a process
called carbon fixation.
The process of photosynthesis can be expressed by the following word equation and chemical equation.

Carbon dioxide + Water  Glucose + Oxygen


6 CO2 + 6 H2O  C6H12O6 + 6 O2

In this lab, you will be using leaf disks, to assay the net rate of photosynthesis under various lighting conditions. Leaf disks
normally float, however when the air spaces are infiltrated with carbon dioxide, the overall density of the lead disk increases and the
leaf disk sinks. When sodium bicarbonate is added to the water, the bicarbonate ion acts as a carbon source for photosynthesis
causing the leaf disks to sink. As photosynthesis proceeds, oxygen is released into the interior of the leaf, which changes its
buoyancy causing the disk to rise. Since cellular respiration is taking place at the same time in the leaf, the oxygen generated by
photosynthesis is consumed. As a result, the rate that the disks rise is indirectly proportional to the net rate of photosynthesis.

Hypothesis:

Materials:
Sodium Bicarbonate , Liquid Soap , Syringe (20 ml), Leaf, Hole Punch, Timer, Light Source , Stir sticks
Plastic cups, Beakers, Tin Foil

Procedure:
Label 3 plastic cups with the following: Light, Ambient Light and Dark
Mix a small scoop (1/8 tps) sodium bicarbonate into 300 ml of water in a beaker
Add 1 drop of soap to the bicarbonate solution. If your solution generates excessive suds, add more water and bicarbonate.
Hole punch 30 uniform leaf disks in texture and thickness avoiding major leaf veins (10 for each trial)
Remove the plunger of the syringe and place 10 leaf disks in the syringe barrel
Replace the plunger being careful not to crush the leaf disks. Push on the plunger until only a small volume of air and lead disk
remain in the barrel
Draw a small volume of the sodium bicarbonate solution into the syringe. Invert the syringe and tap the syringe to suspend the leaf
disks in the solution.
Push the plunger removing as much air a possible from the syringe.
Hold a finger over the syringe opening and draw back on the plunger to create a vacuum. Hold this for 10 seconds while swirling
the syringe to further suspend the leaf disks in solution.
Let off the vacuum and repeat step 8 if needed 2-3 more times until all leaf disks sink. If leaf disks do not sink, add more soap to the
bicarbonate solution and
Pour the disks and solution into the labeled cup
Add the bicarbonate solution until the cup is ½ full
Place the cup under the corresponding light conditions and begin timing
Record the number of floating disks at the end of each minute in the provided table below.
Gently swirl the disk with a stir stick to dislodge any that are stuck to each other or on the side of the cup
Repeat steps 5 – 15 for the other lighting conditions.
For dark trial, wrap a beaker with tinfoil to cover the plastic cup when not taking observations. For the light trial, place the plastic
cup under the lamp.

Predictions:
How will the different light conditions affect the rate of floating leaf disks?
Observations:
Light Ambient Light Dark
Time (min) # of leaf disks Time # of leaf disks Time # of leaf disks
floating (min) floating (min) floating
1 1 1
5 5 5
10 10 10
15 15 15
20 20 20
25 25 25
30 30 30
35 35 35
40 40 40
45 45 45
50 50 50
55 55 55
60 60 60

Qualitative Observations:

Post Lab Analysis:

Graph your results for all three trials in the graph paper below. Label the graph, both axes and provide a legend to distinguish each
trial.

What was the role of the sodium bicarbonate in this experiment?

Which trial resulted in all the leaf disks floating the fastest? Explain.

Explain the process that caused the leaf disks to rise.

If the leaf disks were boiled, what kind of result would you expect? Explain.
10
How does light intensity affect the rate of photosynthesis?
# of leaf disks floating

8
The same experiment was conducted where 10 leaf disks were
placed in a sodium bicarbonate solution and placed in the 6
light. Every minute, the number of floating disks were
counted and recorded. After 14 minutes, the leaf disks were 4

moved into the dark and the number of floating disks were 2
recorded every minute. Below is a graphical representation of
the data. 0
0 5 10 15 20 25 30 35 40 45 50
Time (minutes)

Why did the leaf disks begin to sink after being placed in the dark?

What could the rate of leaf disks sinking be correlated to?


Using your data, what can you deduce about the intensity of light used to obtained the data in the graph?

Lab Report
Your lab report must be typed, font size 12 in an easily readable font.
It is to be written with the following 6 components, in the following order:
1. Title: Should be relevant to the experiment
2. Introduction: This may include some background information and context. It includes the hypothesis that is tested by the lab.
3. Methods: This is a step-by-step procedure of the experiment IN YOUR OWN WORDS. Do not merely copy what is written in
your lab procedure.
4. Results: The data from your experiment is presented. It can be in table or graph form. It must be labeled clearly, using accurate
units.
5. Analysis: This is explanation of your thought process as you reasoned why the initial hypothesis was correct or why it was
incorrect. Use your answers to the Post Lab Analysis questions to help write this part of the report.
6. Conclusion: This summarizes your data and states the conclusion (do you think your hypothesis was incorrect or correct? How
would you change your hypothesis, if at all?) Write about how this experiment is applicable to daily life.

2.Separation of Plant Pigments Using Chromatography

Paper chromatography is a useful technique in the separation and identification of


different plant pigments. In this technique, the mixture containing the pigments to
be separated is first applied as a spot or a line to the paper about 1.5 cm from the
bottom edge of the paper. The paper is then placed in a container with the tip of the
paper touching the solvent. Solvent is absorbed by the paper and moves up the
paper by capillary action. As the solvent crosses the area containing plant pigment
extract, the pigments dissolve in and move with the solvent. The solvent carries the
dissolved pigments as it moves up the paper. The pigments are carried along at
different rates because they are not equally soluble. Therefore, the less soluble
pigments will move slower up the paper than the more soluble pigments. This is
known as developing a chromatogram.

Paper chromatography is useful for identifying unknown compounds - often used in crime scene investigations to match ink,
lipstick, or colored fibers. There are many examples of chromotography at youtube.com. This set-up shows two different pen inks.

Purpose: To identify plant pigments by separation and isolation of the pigments using thin layer paper chromatography.

The distance traveled by a particular compound can be used to identify the compound. The ratio of the distance traveled by a
compound to that of the solvent front is known as the Rf value; unknown compounds may be identified by comparing their Rf's to
the Rf's of known standards.

Rf equation: Rf = distance traveled by compound


distance traveled by solvent

1. Cut a strip of coffee filter (or filter paper). Draw a horizontal line with a pencil (not pen) about half an inch from the bottom.
Place a spinach leaf on the line and roll a penny over it so that you get a line of green pigment on the filter. Using a different part of
the leaf, roll the penny again over the same line. Repeat this process until the line is fairly dark.
2. Put about an inch of acetone in the beaker (isopropyl alcohol will also work.)
3. Tape the top of the coffee filter strip to a pencil and balance the pencil across the top of the beaker. See the image below for the
set-up.
4. It is very important that the bottom of the filter strip is in the acetone, but the green spot is not in the liquid. If the acetone touches
the spot directly, the pigment will just dissolve away.

5. Observe what happens to the liquid in the beaker and the spot on the filter paper. Results will take about 20 minutes.

6.Optional: Repeated using other pigments - try lipstick, felt tip pens, skittles candy..etc
Analysis

1. Assign a band number for each pigment band - you should see greens, yellows, oranges..etc.

Band Color Plant Pigment Distance (mm) Rf (use formula)

Yellow to Yellow-orange Carotene

Yellow Xanthophyll

Bright Green to Blue


Chlorophyll a
Green

Yellow Green to Olive


Chlorophyll b
Green