RESPIRATION—PEA LAB
Introduction:
Aerobic cellular respiration is the process by which cells extract energy from organic
molecules. Glucose is the most common molecule used. It is oxidized according to the
equation
C6H12O6 + 6O2 6CO2 + 6H2O + energy
There are several means by which the respiration of an organism can be demonstrated. A
few of these include the quantity of glucose used, the amount of carbon dioxide released,
and the volume of oxygen consumed. The measurement of oxygen consumption is the
most common. In this exercise you will explore respiration in plants and animals by
measuring oxygen utilization.
Various methods can be used to measure respiratory rates (respirometry). Most
techniques involve instrumental, chemical, or volumetric measurements of respiratory
gases (i.e. CO2 and O2). In this exercise you will attempt to measure volumetric (gas
volume) changes in oxygen by using simple respirometers as shown in Fig. 1.
The respiratory chambers represent closed systems. Any variation in the volumes
(pressure) within the system in which an organism is respiring represents the difference
between oxygen consumption (decreases volume) and carbon dioxide production
(increases volume). Since carbon dioxide is removed by the absorbent KOH, any
volumetric changes within the system is due to oxygen consumption.
Materials Needed: Figure 1. Seed Volumeter
Respiration—Pea Lab 9-1
Procedure:
1. Obtain all the materials shown in Fig. 1. All the tests will be made using this
apparatus.
2. Use 20, room temperature, germinating pea seeds. This is your respirometer.
Obtain weight of germinated (living) peas: _________________________________
3. Secure a loose wad of cotton over the peas, do not pack tightly. Then add 1 cm thick
layer of potassium hydroxide (KOH) pellets over the cotton. Make sure the pellets do
not contact the peas! Do not touch allow the KOH pellets to touch your skin as
they are very caustic.
4. Prepare another test tube the same way, except use 20 boiled pea seeds. This is your
control tube.
5. Fit a stopper, with attached capillary and vertical tubing, snugly into each tube.
Secure the test tubes in a vertical position with the utility clamps and support stand.
Make sure the horizontal portion of the capillary tube is level.
6. Use an eyedropper to add enough colored water to the end of each capillary tube so
that 1 cm of the colored water will be drawn in. Make sure the clamp is open.
7. Allow the system to equilibrate for 5 minutes. Mark the position of the outer
meniscus of the colored water on the tube with a wax pencil.
8. Tightly clamp off the rubber sleeve and allow the peas to respire for 15–20 minutes.
After this time, mark the outer meniscus again and measure the distance the colored
water drop has traveled from the 1st mark in mm.
9. Record the results in the table below. If the colored water moves rapidly, you should
stop before the colored water reaches the bend in the tube and record that time.
10. If a second run is desired, release the clamp on the rubber sleeve and readjust the
colored water by tilting the capillary tube, returning the colored water drop to the
outer end of the tubing. Perform a second run using the same peas and record your
results in the table. Erase the marks then remark the new colored water position with
a wax pencil.
Respiration—Pea Lab 9-2
Name___________________________
Respirations—Pea Lab Report Sheet
Table 1. Pea Seed Analysis
* r = radius of inside diameter of tube in mm.
A B C D E
AVERAGE O2
RESPIROMETER CONTROL CORRECTED VOLUME O2 CONSUMED
(Germinated peas) (Boiled peas) DISTANCE CONSUMED
Measured Measured Column D
Distance in mm Distance Column A — Column C divided by
in mm Column B
times π r2* gm of peas &
time (10 minutes)
TRIAL
1 mm3 O2
mm mm mm 3
mm O2 g x min
TRIAL
2 mm3 O2
3
mm mm mm mm O2 g x min
1. Why was it necessary to remove CO2 from the test tubes? ______________________
____________________________________________________________________
____________________________________________________________________
2. What was the object of using boiled peas? __________________________________
____________________________________________________________________
____________________________________________________________________
3. Why did the colored water column move toward the respirometer? _______________
____________________________________________________________________
____________________________________________________________________
Respiration—Pea Lab 9-3
