Ethan Chang

Majoos
November 28, 2013
Cell Respiration


Introduction

All cells require energy to perform chemical reactions that are essential in their
survival. Aerobic cellular respiration is the main source of energy for most organisms.
Animals or other single-celled organism sometimes rely on anaerobic respiration when
oxygen is not present. But in this experiment, we will mainly focus on aerobic cellular
respiration, which is a sophisticated process that needs help from various enzymes and
involves many intermediate steps to ultimately gain energy by oxidising glucose. Heres an
overall equation for aerobic cellular respiration regarding reactants and products:
C6H12O6 + 6O2 > 6CO2 + 6H2O + energy
Our goals in this experiment are to prove the existence of cell respiration by
measuring the consumption of oxygen by seeds and also to compare the respiration rates
of germinating and dry pea seeds at two different temperatures. The experimental peas
are put into a vial, which is then sealed with a rubber with a pipet inserted in it. The vial is
later on submerged in water. If the peas do respire, then water will rush in the vial through
the pipet.
The gas laws, especially Avogadros Law, are important conceptions in this
experiment. Avogadros Law states that equal volumes of all gases, at the same
temperature and pressure, have the same number of molecules, which can be written as
V1/n1 = V2/n2. According to this law, the gas volume of the respirometers should remain the
same even if the peas respire because there is no net change in moles of gas as shown in
the equation above. However, for testing purposes, potassium hydroxide (KOH) is used to
interfere this equilibrium by reacting with CO2 to form solid potassium carbonate (K2CO3).
Since the equilibrium established previously is altered, the volume of gas in the
respirometer will decrease and water will replace that blank space. From that, the rate of
cellular respiration can be easily calculated.

Procedure

In this experiment, rst, two trays of water of different temperature (one at room
temperature, the other at preferably 10) are prepared to provide conditions for the
respirometers. A sheet of pater is advised to put in the bottom of the container because it
will make the pipet easy to observe. Next, to assemble two identical sets of three
respirometers, rst an absorbent cotton with potassium carbonate is placed in the bottom
of each vial and a nonabsorbent cotton is placed on the top of it to prevent potassium
carbonate from contaminating the seeds. After that, for one set, 25 germinating peas are
dropped in one vial. 25 dry peas and also glass beads are dropped in another vial to equal
the volume of the rst vial because dry peas are usually much smaller than germinating
peas. The last respirometer is lled up with glass beads to serve as negative control.
After all the respirometers are prepared, they are being immersed in the trays of
water for a few minutes with their tips resign on the edge in order to reach thermal
equilibrium. After thermal equilibrium is reached, the whole respirometer is submerged in
the water. Initial volume of the vial is observed and recorded down by reading the scale.
This step will be repeated every ve minutes for 20 minutes.

Data











Table 1: Respiration of Peas at Room Temperature
Respirometer 1 Germinating
Peas
Respirometer 3
Dry Peas + Beads
Respirometer 3
Beads Only
C

Time
(min)
V of
Pipet
"V
Corrected
"v
V of Pipet "V
Corrected
"v
V of Pipet "V
22 0 10 - - 10 - - 10 -
22 5 8.5 1.5 0.5 9.3 0.7 -0.3 9 1.0
22 10 8.1 1.9 0.9 9 1.0 0.0 9 1.0
22 15 7.7 2.3 1.3 9 1.0 0.0 9 1.0
22 20 7.2 2.8 1.8 8.8 1.2 0.2 9 1.0
Table 2: Respiration of Peas at Colder Temperature
Respirometer 1 Germinating
Peas
Respirometer 3
Dry Peas + Beads
Respirometer 3
Beads Only
C

Time
(min)
V of
Pipet
"V
Corrected
"v
V of Pipet "V
Corrected
"v
V of Pipet "V
10 0 10 - - 10 - - 10 -
10 5 7.3 2.7 0.2 7.8 2.2 -0.3 7.5 2.5
10 10 7.2 2.8 0.3 7.9 2.1 -0.4 7.5 2.5
10 15 7.2 2.8 0.6 8.3 1.7 -0.5 7.8 2.2
10 20 7.4 2.6 0.8 8.7 1.3 -0.5 8.2 1.8
Graphs



Analysis Questions

1. Write two hypotheses that this experiment is designed to test.

a. If equal volumes of germinating peas, dry peas and glass beads are allowed to
perform cellular respiration in the vials under the same pressure and temperature, the
gas volume of respirometer that contains germinating peas will decrease the most.
b. If germinating peas and nonterminating peas are allowed to respire in the vials at room
temperature or 10C, the volume of gas of respirometers that is placed under room
temperature will decrease more.

Cell Respiration of Peas
C
o
r
r
e
c
t
e
d

"
V
7
7.15
7.3
7.45
7.6
7.75
7.9
8.05
8.2
8.35
8.5
8.65
8.8
8.95
9.1
9.25
9.4
9.55
9.7
9.85
10
Time
0 5 10 15 20
Germinating Peas at 22 Dry Peas at 22 Germinating Peas at 10 Dry Peas at 10
2. In this experiment, you measured the change in volume of the gas inside the
respirometers. The general gas law describes the state of a gas under given
conditions.
pV = nRT
where p = pressure of the gas
V= volume of the gas
n = kmoles (number of molecules) of gas
R = universal gas constant [8314 joules/(kmole)(K)]
T = temperature of the gas in K
Since you have been measuring change in volume, restate the general gas law to solve for
volume:
V = nRT/p
Using the general gas law and your experience in this lab give the variables that had to be
controlled for your data to be valid. State the controls used for each variable and any
means used to correct for the inuence of a variable(s).

According to the gas law, if were measuring the change in volume caused by the change
in amount of gas, the other variables must remain constant. R is an universal gas
constant. As for pressure, since the whole experiment is being conducted under standard
atm from the beginning to the end, it should affect the result. Temperature is the key
variable that requires efforts to maintain. Room temperature shouldnt change a lot for 20
minutes. For the second tray of water, ice is put in to cool it down to 10C. If the
temperature rises up by observing the scale on thermometer, more ice will be put in to
maintain a cold temperature. Lastly, it is also important to make sure the initial volume of
germinating peas, dry peas, or glass beads is same for every respirometer. That way, a
more accurate change of amounts of gas can be measured.

3. Which of the respirometers (1, 2, or 3) serves as a negative control? Explain your
answer.
In this experiment, respirometer 3 serves as a negative control. Respirometer 3 is the one
where no change of volume is expected to happen. It is there for the observer to determine
whether cellular respiration accounts for the change in gas volume or the gas volume
simply decreases naturally. That is the reason why "V for respirometer 1 and 2 is used to
subtract "V of respirometer 3 to gain corrected "V, which is a more accurate result.

4. In reference to the general gas law, and assuming your control measures worked, a
change to which of the variables led to the observed change in volume (Corrected "V
in Table 1 and 2)? Explain your answer.
Assuming all the other variables remain constant, change in moles of gas is the reason for
the change in volume, as the Avogadros Law states that volume of gas is directly
proportional to the number of molecules under the same temperature and pressure.

5. Using your graph and data tables, summarise your ndings, comparing results from
respirometers 1 and 2, and results obtained at room temperature vs. results at the
colder temperature. Speculate as to the causes(s) of any differences between the
treatments.
My two hypotheses are proven correct in accordance with my graph and data tables. The
volume of gas decreases the most in respirometer 1 compared to respirometer 2 and 3
because germinating peas are more active in cellular respiration than dry peas. Also, the
gas volume in respirometers at room temperature clearly decrease more than
respirometers at colder temperature because room temperature not only allows enzyme to
be more active but also increase kinetic energy of molecules.


6. From your graph, calculate the rate of oxygen consumption for each treatment:
a. germinating seeds at room temperature = 0.09 mL/min
b. Germinating seeds at colder temperature = 0.04 mL/min
c. dry seeds at room temperature = 0.01 mL/min
d. dry seeds at colder temperature = -0.025 mL/min


Conclusion

Most part of our date from the experiment seems correct except the result from
respirometer 2 at colder temperature, which has a negative rate of oxygen consumption.
The reason behind this minor error could be that respirometer 2 was somehow interfered
during the process, or respirometer 3, which serves as a negative control, had been picked
up from the water bath so that some air came out of the pipet and therefore water would
rush in when the respirometer is put back at the bottom of the tray. The second
explanation sees more logical because unlike respirometer 3 at room temperature, which
only lost one mL of air, the change of gas volume in respirometer 3 at colder temperature
is 2.5 mL, which is abnormal. Despite these minor technical errors, our data sill show a
generally correct result that accurately explains and measures oxygen consumption in
cellular respiration.