AP Biology

Investigation 6: Cellular Respiration READ ALL OF THIS!

Essential Question: What factors can affect the rate of cellular respiration?

Background

Living systems require free energy and

matter to maintain order, to grow, and to
reproduce. Energy deficiencies are not
only detrimental to individual organisms,
but they cause disruptions at the
population and ecosystem levels as well.
Organisms employ various strategies that
have been conserved through evolution to
capture, use, and store free energy.
Autotrophic organisms capture free
energy from the environment through
photosynthesis and chemosynthesis,
whereas heterotrophic organisms harvest free energy from carbon compounds produced by
other organisms.

The process of cellular respiration harvests the energy in carbon compounds to produce ATP
that powers most of the vital cellular processes. In eukaryotes, respiration occurs in the
mitochondria within cells. If sufficient oxygen is available, glucose may be oxidized completely in
a series of enzyme-mediated steps, as summarized by the following reaction:

C6H12O6 + 6O2(g) 6CO2(g) + 6H2O + energy

More specifically, C6H12O6 + 6O2 6CO2 + 6H2O + 686 Cal/mole of glucose

During cellular respiration, two gases are changing in volume. Oxygen gas is being consumed
by the respiring cells and carbon dioxide gas is diffusing out of the cells. The respirometer,
therefore, has to be able to deal with two simultaneously changing gas volumes. This is
accomplished by introducing potassium hydroxide into the device. KOH absorbs carbon dioxide,
following this equation:

CO2 + 2KOH K2CO3 + H2O

Potassium carbonate (K2CO3) is a solid precipitate. Any CO2 produced is immediately converted
from a gas to a solid and is therefore no longer governed by gas laws. This allows the
respirometer to measure only one variable, the consumption of oxygen gas by living cells.

Adapted from
The Biology Corner: Investigation What affects cellular respiration?
College Board AP Biology Lab 5 (Dirty Dozen Version)
And College Board AP Biology Investigation 6
AP Biology
Investigation 6: Cellular Respiration READ ALL OF THIS!

Learning Objectives

To learn how a respirometer system can be used to measure respiration rates in plant
seeds or small invertebrates, such as crickets (if you want to capture them and bring
them in, that is up to you).
To design and conduct an experiment to explore the effect of certain factors, including
environmental variables, on the rate of cellular respiration
To connect and apply concepts, including the relationship between cell structure and
function (mitochondria); strategies for capture, storage, and use of free energy; diffusion
of gases across cell membranes; and the physical laws pertaining to the properties and
behaviors of gases

Safety Precautions

You must wear safety goggles or glasses, aprons, and gloves during this investigation(s)
because KOH (or the alternative, NaOH in Drano) is caustic.

Pre-Lab Questions

These items should be completed in order to design an appropriate experiment.

1. What is the difference between a non-germinating vs. germinating seed with respect to
respiration? A non-germinating seed tends to respire less than germinating seeds. This
is because the germinating seeds need more energy to grow and develop cells.

2. What are two primary ways to quantify the rate of cellular respiration? B
y the amount of
oxygen gas consumed and the amount of carbon dioxide gas released.

3. Why is it necessary to correct the readings of the respirometers containing seeds with
the readings taken from respirometers containing only glass beads? Your answer should
refer to the concepts derived from the general gas law, PV = nRT. The readings of
respirometers should be corrected due to the fact that the volume of gas, fluctuates,
indicating that the amount of oxygen present likely changes as well. This way, it reduces
chance of error.

4. How can you reduce variability in your samples? Should you use the same number of
seeds, same mass, same volume, or something else? Justify your reasoning. Y ou
should use the same volume of items in each sample, this way the amount of
fluctuations that occur with the volume of oxygen gas doesnt impede the accuracy of the
results. Essentially, they have the same baseline volume.

Materials*

Safety goggles, aprons, and gloves 25 non-germinated beans

4 respirometers Glass beads
25 germinated beans Vasoline

Adapted from
The Biology Corner: Investigation What affects cellular respiration?
College Board AP Biology Lab 5 (Dirty Dozen Version)
And College Board AP Biology Investigation 6
AP Biology
Investigation 6: Cellular Respiration READ ALL OF THIS!

Styrofoam cup Non-absorbent cotton

Thermometer Absorbent cotton
Water, about 200 ml Potassium Hydroxide solution
Food coloring Timer

*Some materials may need to be added/altered due to the nature of your designed inquiry
set-up.
Procedure

You will conduct a control experiment using 3 respirometers at the same temperature. You will
have at least one additional respirometer for your inquiry trial. The purpose of the beads is to
ensure that each respirometer is uniform in volume and allows you to account for pressure
conditions in the lab environment. As oxygen is used up by the respiring seeds, the gas
pressure inside the respirometer will decrease and the food coloring will flow into the pipet down
its pressure gradient.

ASK

Your inquiry respirometer(s) should investigate a way to alter respiration rate that allows you to
compare it to at least one of your controls. In addition to the materials required for the control
experiments, typical inquiry experiment supplies include the other seed types, hot plates, ice (if
asked for), HCl, and NaOH. Some materials you may need to bring in yourself. If you need
something else, I will try to get it for you as long as you can justify needing it. Get your own bugs
if you want some!

Determine the following information for your inquiry lab:

1. What is the independent variable? Temperature

2. What i s the dependent variable? Amount of oxygen consumed (distance food coloring
travels; the more oxygen has been consumed increases with the distance food coloring
travels since it only moves with reduction in gas pressure)
3. What is your hypothesis? Provide an explanation for your hypothesis. I f a higher
temperature is used, then the amount of oxygen consumed will increase. This is
because the amount of kinetic energy present in heat will help to increase the movement
of molecules, thus allowing for more reactions to occur. This in turn, increases the rate at
which oxygen is consumed, since it is more likely to be involved in these processes.
4. What variables will be held constant? Starting volume, amount of seeds, amount of
water in water bath, amount of KOH, type of seed, temperature of water bath.
5. What additional materials will you need to conduct the lab? Beakers, Graduated
Cylinders, hot plate.
a. What will YOU be responsible for bringing on lab day? A ll materials will be
provided by the teacher.

Your inquiry design must be approved of before moving on to the next phase of the EDP.

Adapted from
The Biology Corner: Investigation What affects cellular respiration?
College Board AP Biology Lab 5 (Dirty Dozen Version)
And College Board AP Biology Investigation 6
AP Biology
Investigation 6: Cellular Respiration READ ALL OF THIS!

IMAGINE & PLAN

You will be writing your own materials list and procedure with your lab group (Be sure to follow
the Formal Lab Report Guidelines). The information below should be included in your
procedure

1. You will set up at least 4 respirometers as shown below. Be sure to seal the stopper
against the vial and around the pipet with Vaseline!
2. For the control experiments, place all three respirometers into a styrofoam coffee cup
and cover with water.
3. Measure and record the temperature of the water.
4. Allow the system to equilibrate for approximately 5 minutes.
5. Place a drop of food coloring on top of each pipet to start the experiment.
6. For each respirometer you have, you should record the temperature of the water bath,
the total distance the food coloring has moved, and the distance it moved in the time
interval (current volume minus the previous volume) every 5 minutes for 25 minutes.

Respirometer Contents

1 Glass beads

2 Non-germinating seeds

3 Germinating seeds

4+ Your choice

Adapted from
The Biology Corner: Investigation What affects cellular respiration?
College Board AP Biology Lab 5 (Dirty Dozen Version)
And College Board AP Biology Investigation 6
AP Biology
Investigation 6: Cellular Respiration READ ALL OF THIS!

The image to the right shows The image above provides the general
how a coffee cup can setup for your control respirometers.
function as a water bath to The non-absorbent cotton protects
maintain a constant your organisms from the KOH at the
temperature. A drop of food bottom. Do not mix up your cotton!!!
coloring is added to the tip of
the pipet AFTER the system Available seed types:lima beans, pinto
has equilibrated for beans
approximately 5
minutes.

Lab Group Procedure:

*I must approve of your lab procedure before you are allowed to conduct the lab!

1. Put on gloves, goggles, and aprons.

2. In all of the vials, soak absorbent cotton with KOH solution.
3. Cover with dry non-absorbent cotton.
4. Set up at least 6 respirometers
5. Two respirometers should have 25 germinating field peas.
6. Two respirometers should have 25 non-germinating seeds and glass beads so that the
volume matches the volume of the respirometers with germinating field peas.
7. Two respirometers should have enough glass beads so that the volume matches the
volume of the respirometers with germinating field peas..
8. Cap each vial with a stopper fitted with a pipette, tip pointing outward, sealing it with the
vaseline.
9. Place in 6 styrofoam cups 200ml of water. Cool down three cups to 10oC by adding ice
cubes. Cool down another three cups to 25oC by adding ice cubes to them as well. Allow
five minutes for the respirometers to equilibrate.
10. Take an initial reading from each respirometer
11. Record the temperature of the water bath for each respirometer.
12. Put one drop of food coloring on top of each pipet to begin experiment.
13. Once experiment has begun, record the distance the food coloring has moved (current
volume subtracted from previous) in 5 minute intervals for 20 minutes.

Adapted from
The Biology Corner: Investigation What affects cellular respiration?
College Board AP Biology Lab 5 (Dirty Dozen Version)
And College Board AP Biology Investigation 6
AP Biology
Investigation 6: Cellular Respiration READ ALL OF THIS!

CREATE

You should be prepared to set up the three control experiments AND your inquiry experiment to
run almost simultaneously. The experiments themselves need to run for 30 minutes AFTER
setup, so be able to prepare your respirometers quickly (by knowing how they should be
packed) and assign roles for each of your group members before lab day.

Create a data table below.

Germinating Dry Peas with Germinating Dry Peas with

Time X Peas (10oC) Beads (10oC) Peas (24oC) Beads (24oC)
0 0 0 0 0
5 0.04 0 0.08 0
10 0.06 0 0.18 0.01
15 0.09 0 0.29 0.02
20 0.1 0.01 0.4 0.02

Insert a graph of your data. Be sure to include a graph title, axis labels, and a key.

Adapted from
The Biology Corner: Investigation What affects cellular respiration?
College Board AP Biology Lab 5 (Dirty Dozen Version)
And College Board AP Biology Investigation 6
AP Biology
Investigation 6: Cellular Respiration READ ALL OF THIS!

Yellow line- Germinating Peas (24oC)

Blue line- Germinating Peas (10oC)

Green line- Dry Peas with Beads (24oC)

Red line- Dry Peas with Beads (10oC)

IMPROVE

1. How was the rate of cellular respiration affected by the experimental variable(s) you
chose as compared to the control(s)? The rate of cellular respiration with the
experimental variables of germinating peas at 24oC (0.02 ml O2/ min) was higher in
comparison to the control group, glass beads at 24oC (0.001 ml O2/min).

2. Was your initial hypothesis about the effect of your factor on the rate of cellular
respiration supported? Why or why not? My initial hypothesis about the effect of
temperature on the rate of cellular respiration was supported by the above results. The
peas tended to respire more at higher temperatures in comparison to lower
comparisons. This is especially evident between the Germinating Peas at both 24oC and
10oC, which showed the greatest contrast. The Germinating Peas at 24oC had a rate of
respiration of 0.02 ml O2 /min while the Germinating Peas at 10oC had a rate of
respiration of 0.001 ml O2/min. The higher rate of respiration seen at the higher
Adapted from
The Biology Corner: Investigation What affects cellular respiration?
College Board AP Biology Lab 5 (Dirty Dozen Version)
And College Board AP Biology Investigation 6
AP Biology
Investigation 6: Cellular Respiration READ ALL OF THIS!

temperature indicates that temperature is a big factor when trying to assess the
efficiency of cellular respiration, with higher temperatures leading to higher rates.

3. What were some challenges you had in performing your experiment? Did you make any
incorrect assumptions? Some challenges we had performing our experiment was
managing our time properly. We spent a lot of time setting up the experiment as
opposed to doing the actual experiment, which is reflected by our small amount of data.
We also were originally going to use an insect to compare respiration rates between
organisms. However, we incorrectly assumed that it would be easy to put the bug in the
respirometer. It was not. As a result we had to change our comparison groups to using
another bean type in order to perform the experiment.

We also, as a class, had to use sample data from the lab instruction book, in order to
answer analysis questions. This is due to the fact that the data we collected wasnt enough to
gather a strong conclusion.

4. Were you able to perform without difficulty the mathematical routines required to analyze
your data? If not, what calculations were challenging or required help from your
classmates or teacher? Yes we were able to perform the mathematical routines required
to analyze our data. The chart above shows a clear relationship between the the amount
of oxygen consumed with respect to time. These variables can be used to find the rate of
oxygen consumption, which a necessary component of cellular respiration. Simply taking
the slope of the graph (change in y/change x) and assigning oxygen consumption as the
y and time as the x allows us to find the ratio ml O2/ min, or the rate of oxygen
consumption over time. Any values with that particular unit acted as a base of
comparison at which to analyze, which made the analysis aspect of the experiment
much simpler.

Adapted from
The Biology Corner: Investigation What affects cellular respiration?
College Board AP Biology Lab 5 (Dirty Dozen Version)
And College Board AP Biology Investigation 6