Mary Ward

AP Biology

Schulte

Experiment #4

Date of Experiment: 2/14/18

Title:
Measuring and calculating the rate of oxygen consumption in germinating seeds using
respirometry techniques
 Abstract:

The purpose of this experiment was for students to observe and measure the rate of
oxygen consumption (cellular respiration) in germinating mung beans compared to non-
germinating mung beans and glass beads in a 10 degrees Celsius ice bath. The hypothesis for this
experiment is that students should expect to see a higher rate of oxygen consumption if
germinated beans are present rather than the non germinated beans and glass beads.

In this experiment three respirometers were used. First three graduated cylinders were
used to determine what should go in each respirometer. The first graduated cylinder’s contents
will later be placed in respirometer one, the second graduated cylinder will correspond with
respirometer two, and the third graduated cylinder will correspond with respirometer three. The
first graduated cylinder for respirometer one was filled with 50 mL of water along with 25
germinating mung beans, equivalent to 3 mL, the final volume equaling 53 mL. The second
graduated cylinder for respirometer two was also filled with 50 mL of water along with 25 non-
germinating mung beans and enough glass beads to be equivalent to 53 mL. The third graduated
cylinder for respirometer three was again filled with 50 mL of water and only glass beads to
equal 53 mL total. All three graduated cylinders were then drained and the solid contents
(germinating and non-germinating mung beans and glass beads) were placed on a paper towel to
dry. Absorbent cotton that was drenched in 15% KOH to activate the cellular respiration process
was placed at the very bottom of all three respirometer along with non-absorbent cotton on top of
the absorbent cotton to block the airflow. Now students will place the appropriate beans and
glass beads into the correct respirometer. Masking tape was then used to prevent airflow from
entering the respirometers because the experiment would fail if air entered the respirometers
since carbon dioxide would go from a human source instead of the beans and beads. This
experiment was supposed to be done at two different water temperature but in this version only
10 degrees Celsius water was used and not room temperature. So only the oxygen consumed was
measured and not difference in temperature. The independent variable for this experiment was
the amount of beans and/or beads placed in each respirometer. The dependent variable for this
experiment was the rate of oxygen consumption.

Materials and Equipment Used:

● 50 germinating seeds
● 50 dry seeds
● 100 plastic beads
● 3 respirometer vials
● Weights for vials
● 3 stoppers
● 1 mL graduated pipettes
● Sealant (Vaseline)
● Absorbent cotton
● Non Absorbent cotton
● 1 round wood stick
● 3 pieces of paper towel
● Marking pen
● Water bath
● Ice
● 100 mL graduated cylinder
● Thermometer
● Masking tape
● Stopwatch or clock
● Water
● Dropper bottle of 15% KOH

Procedure and methods:

1. A 10 degrees Celsius bath should be set up immediately to allow for time to adjust the
temperature of the bath. (Add ice to attain the 10 degrees Celsius)
2. Respirometer 1: Obtain a 100 mL graduated cylinder and fill it with 50 mL of water.
Place 25 germinating peas into the graduated cylinder. Measure the amount of water that
was displaced (which is equivalent to the volume of the peas) and record this number.
3. Respirometer 2: Refill the graduated cylinder until it has 50 mL of water. Place 25 dried
peas (they are dormant, and not germinating) into the graduated cylinder and then add
enough glass beads to reach a volume equivalent to the of the germinating peas in
Respirometer one. Remove the peas and beads and place them on a paper towel. These
beads will be used in Respirometer two.
4. Respirometer 3: Refill the graduated cylinder until it has 50 mL of water. Fill it with
glass beads alone until the volume is equivalent to the volume of the germinating peas in
Respirometer one. Remove these beads and place them on a paper towel. These beads
will be used in Respirometer three.
5. To assemble the respirometers, obtain 3 vials, each with an attachable stopped and
pipette. Make sure that he vials are dry in the inside. It is important that the amounts of
cotton and KOH are the same in each vial. Place a small wad of absorbent cotton in the
bottom of each vial and,using a dropper, saturate the cotton with 15% KOH. Place a
small wad of dry non absorbent cotton on the top of the KOH soaked absorbent cotton.
6. Place the first set of germinating peas, dry peas and beads, and beads in all three vials.
 7. Insert the nontrapered end of one pipette into the upper surface of one of the rubber
stoppers. It should fit tightly. Place a layer of sealant around the junction between the
pipette and the stopper so that no air can escape.
8. Wrap parafilm tightly around each stopper (where it enters the vial and where the pipette
is inserted). This is necessary to ensure against any leaks. Any leaks will cause the
experiment to fail.
9. Place a strip of masking tape across the narrow width of the water bath, approximately ⅔
of the way from one end.
10. Place all three respirometers into the 10 degrees Celsius water bath so that the pipettes
rest on the masking tape prop. Start a timer for 7 minutes - this is the equilibration period
where your respirometers will become the same temperature as the water.
11. At the end of the 7 minutes submerge each of the tubes entirely in the water bath. Some
water will enter the tip of the pipette, but the influx of water should stop fairly quickly. If
it doesn’t stop check the respirometer for leaks.
12. At this point check to make sure you read the pipettes. The air bubble should extend from
the main chamber up the tube of the pipette. The pipette may need to be rotated so that
you can see the numbers. If your respirometers float, you may need to weight them.
13. Record the water level in each of the three pipettes onto the data table at time interval 0.
14. Then record the position of the water in each pipette at the end of 5, 10, 15, and 20
minutes in your data table.

Results:

Respirometer Preparation
Respirometer Temperature Contents

1 10 degree Celsius Germinated seeds

2 10 degree Celsius Dry seeds + Beads

3 10 degree Celsius Beads

This data table shows what should be placed into each of the three respirometers and
what temperature water the respirometer then need to be placed in.

Volume within each respirometer

 Respirometer 1 Respirometer 2 (non- Respirometer 3
(germinating seeds) germinating) (beads only)

Initial Volume (mL) 50 mL 50 mL 50mL

Final Volume (mL) 53mL 53 mL 53mL

Volume of 3 mL of seeds 2 mL of seeds (25) 3 mL of beads (34)

beads/seeds 1 mL of beads (17)

This data table shows the initial and final volumes of each respirometer. It also shows the
volumes of the beans and beads along with the amount of beans and beads needed to each the
volumes in the respirometers.

Final Volumes
Germinating seeds volume Non-germinating seeds Beads volume
volume

4.75 mL 4.5 mL 2.5 mL

This data table shows the final volumes that were discovered at the end of the experiment
in each of the three respirometers.

Analysis/Conclusion:

The purpose of this experiment was to observe and measure which form of beans and
beads consume the most oxygen, so it was fulfilled. The germinating beans consumed the most
oxygen because it had produced a volume of 4.75 mL when placed in the 10 degrees Celsius
water. The non-germinating beans produced a volume of 4.5 mL; however, these beans were not
supposed to perform cellular respiration because they don’t produce or recieve gases (oxygen
and carbon dioxide) required to perform the cellular respiration process. This could be a cause
for human error. The glass beads consumed the least of oxygen because it produced a volume 2.5
mL but since the beads are abiotic they should has had a volume of 0 mL since the can’t undergo
the process of cellular respiration. This could also be due to human error.

The hypothesis of this experiment was technically proven correct but since there were
several errors in the experiment the results could have been skewed. The respirometer with
germinating beans did have the highest amount of oxygen consumption so this shows that the
hypothesis is correct. Although the non-germinating beans and glass beads were not supposed to
consume any oxygen both had their volume increase which shows possible human error since
their volumes should have stayed the same.
 There were several sources of error during this experiment that most likely skewed the
results. It is possible that air entered the respirometers allowing other sources of carbon dioxide
in. Another possible explanation for the errors in this experiment is that some germinating beans
got into the wrong respirometer. This experiment should be performed multiple times in the
future to eliminate the chance of human error.

Background Research and Bibliography:

A method of indirectly measuring the rate of cellular respiration in to monitor changes in

temperature since this process is exergonic. Enzymes play a vital role in cellular respiration so if
they are denatured the process is interrupted. Another method of measuring the rate of cellular
respiration is to measure oxygen consumption or carbon dioxide production by using
respirometers, which can measure gas volume changes. In this experiment students measured the
rate of oxygen consumption.

S. V. (n.d.). Does temperature change affect the respiration of yeast. Retrieved March 1, 2018,
from
https://vle.stvincent.ac.uk/2014/pluginfile.php/33446/mod_resource/content/2/Does-
temperature-change-affect-the-respiration-of-yeast-10vsrbd%20(1).pdf