Biochemistry, Osmosis, Cellular Respiration & Photosynthesis Lab Stations

Raina Kim

Station 1: Extracting enzymes from potato tissue and making water baths

Manipulated Variables:
Temperature of each cylinder
Controlled Variables:
Amount and type of potato filtrate
Amount and type of 3% hydrogen peroxide
Time used for the pressure of the graduated cylinder
Responding Variables:
The pressure of each beaker that is manipulated
The colour that it changes to after the solution reacts afterwards
The pH before and after the reaction
Mass of Potato:
This was prepared before the lab, therefore we were unable to measure to mass of the
potato
Materials:
4 x 10 mL graduated cylinders cutting board
100 mL graduated cylinder blender and soft spatula
250 mL beaker cheesecloth or coffee filter
2 x 600 mL beakers thermometer
3 medium test tubes that fit the stopper pH paper or pH probe
from the gas pressure probe hot plate
test tube rack Vernier lab quest and pressure probe
ice with test tube attachment
10 mL of 3% hydrogen peroxide timer / stop watch / chronometer
1 potato balance
knife

Procedure
1. Mass the potato and record.
2. Wash and cut the potato into 4 pieces using the knife and cutting board.
3. Add the potato quarters to the blender with 100 mL of distilled water. Blend until smooth.
4. Pour the liquid through cheesecloth or coffee filter and collect the filtrate in the 250 mL
beaker.
5. Measure 2.0 mL of filtrate into each of 3 test tubes. Place one in a 600 mL beaker half full of
ice water temp: ____ C, place one in a test tube rack at room temperature: _____ C, and
place one in a 60C 70C hot water bath temperature actual temp: __________ C (made
with the hot plate and a 600 mL beaker half full of tap water).
6. Measure 3 x 2.0 mL of 3% hydrogen peroxide into 10 mL graduated cylinder and bring them
to the same temperature as the potato filtrate.
7. Measure with a thermometer and record the temperatures of all three set-ups.
8. For each of the temperature set-ups: monitor the pH of the solution with a pH probe.
 9. Invert the hydrogen peroxide into the test tube containing the potato filtrate with the pH probe
inside of the test tube and start collecting data. Record the initial pH and final pH after 30 s.
10. Repeat with all three temperatures.
11. (If time had allowed we would choose at least two more temperatures and do at least 5 trials
of each temperature).

Table #1: The change of both chemical and physical characteristics for a potato in
varied temperatures
Temperature Initial Pressure Final Pressure Initial pH Final pH
(C) (0.5) (kPa) (0.05) (kPa) (0.05) (0.5) (0.5)

Ice water bath 0.0 90.23 91.00 7.0 6.0

Room 21.5 90.25 92.21 6.5 6.0

temperature

Hot water bath 68.9 90.48 90.23 6.5 6.5

Station 2: diluting, determining the isotonic point of a potato, calculating % change

 Table #2: Initial, final and change in mass of 1cm x 1cm potato cubes, placed in
different sucrose solutions of 0M, 0.20M, 0.40M, 0.60M, 0.80M and 1.00M
Molar concentration Initial Mass of Final Mass of potato % Change in Mass
of sucrose solution potato cubes (g), cubes (g), (0.01g) of potato
(mol/L) (0.01g) cubes(5%)
(0.01mol/L)

0.00 0.69 0.40 -42.0%

0.20 0.69 0.46 -33.3%

0.40 1.03 0.83 -19.4%

0.60 1.37 1.21 -11.7%

0.80 1.08 1.23 13.9%

1.00 0.99 1.23 24.2%

Table #3: The initial, final and % change in mass of five trials of 1cm x 1cm potato
cubes, placed in different sucrose solutions of 0M, 0.20M, 0.40M, 0.60M, 0.80M and 1.00M
Molar Sample 1 Sample 2 Sample 3
concentr
ation of Initial Final % Initial Final % Initial Final %
sucrose Mass Mass Change Mass Mass Change Mass Mass Change
solution of of in Mass of of in Mass of of in Mass
(mol/L) potato potato of potato potat of potato potato of
(0.01m cubes cubes potato cubes o potato cubes cubes potato
ol/L) (g), (g), cubes( (g), cubes cubes( (g), (g), cubes(
(0.01 (0.01 5%) (0.01 (g), 5%) (0.0 (0.01 5%)
g) g) g) (0.0 1g) g)
1g)

0 0.99 1.23 24.2% 1.11 1.36 23% 0.82 0.76 7%

0.2 1.08 1.23 13.9% 1.23 1.29 5% 0.78 0.8 3%

0.4 1.37 1.21 11.7% 1.32 1.13 14% 1.16 1.04 10%

0.6 1.03 0.83 19.4% 0.91 0.69 24% 0.84 0.7 17%

0.8 0.69 0.46 33.3% 1.03 0.67 35% 1.36 0.79 42%

1 0.69 0.4 42.0% 1.04 0.64 38% 0.86 0.61 29%

 Molar concentration Sample 4 Sample 5
of sucrose solution
(mol/L) Initial Final % Initial Final %
(0.01mol/L) Mass of Mass of Change Mass of Mass of Change
potato potato in Mass potato potato in Mass
cubes cubes of potato cubes cubes of potato
(g), (g), cubes(5 (g), (g), cubes(5
(0.01g) (0.01g) %) (0.01g) (0.01g) %)

0 1.06 0.68 8% 0.34 0.43 26%

0.2 0.92 0.59 1% 0.35 0.34 3%

0.4 0.69 0.47 9% 0.34 0.23 32%

0.6 0.76 0.63 38% 0.3 0.24 20%

0.8 0.93 0.93 37% 0.33 0.24 27%

1 0.99 1.15 31% 0.34 0.23 32%

Table #4: The average and standard deviation of the five trials of 1cm x 1cm potato cubes,
placed in different sucrose solutions of 0M, 0.20M, 0.40M, 0.60M, 0.80M and 1.00M

Molar concentration of Average % change in mass Standard deviation

sucrose solution (mol/L) of a potato with isotonic
(0.01mol/L) point (5%)

0 -6% 0.28788

0.2 -25% 0.356191

0.4 -20% 0.102793

0.6 -20% 0.034742

0.8 -21% 0.212225

1 -21% 0.403621

Graph #1: The average % change in mass of a 1cm x 1cm cube of potato in relation to the
sucrose solution of 0M, 0.20M, 0.40M, 0.60M, 0.80M and 1.00M
An isotonic state allows for the free movement of water across the membrane without
changing the concentration of solutes. Therefore, it is where the concentration of the solution is
the same as the concentration of the cell.
The isotonic point is when x=0 in this case where the sucrose solution is at 0 mol/L. This can be
determined through the best fit line where it is at 9%.

Station 3 - Comparing cellular respiration of germinating and nongerminating seeds in a

respirometer
Table #5: Oxygen concentration measurements taken by a labquest, every thirty seconds for
five minutes, of twenty-five germinating beans of the same species, in a 250mL bottle.
Time (s) Oxygen (%), (0.05)

0 16.01

30 15.97

60 15.90

90 15.88

120 15.87

150 15.86

180 15.85

210 15.84

240 15.84

270 15.84

300 15.83

Rate of Cellular Respiration per Seed:

Slope / 25 seeds
Slope = y / x
= oxygen / time
Pt1 = (300s , 15.83%)
Pt2 = (30s, 15.97%)
= -(15.83% - 15.97%) / (300s - 30s)
= 0.0005185185%/s
(0.0005185185%/s) / 25 seeds
0.0000207407
2.1 x 10^-5 %/s/seeds

Sources of Error:
Errors Improvements

When we measured the concentration of the
germinating beans, they were exposed to
sunlight within the classroom. This could have
activated photosynthesis rather than cellular
respiration through the germinating seeds which
have slight evidences of green: which are
cotyledons. (Batty, 1989).
To obtain more accurate results, we could
perform the lab in a secluded area, without light,
where the photosynthesis is unable to take place.
Cellular respiration is the only reaction that we
are looking for, therefore, another controlled
variable would have to be introduced:
sunlight/environment.
 The twenty-five undamaged germinating beans
cannot actually be chosen accurately because the
beans have the capability to die any second
(N.p., n.d.). They could have looked as if they
were alive with green buds, however they could
retain their colour while in the few seconds on
their death. We were unable to truly and
accurately know which beans were still alive
and germinating. If some beans were already
dead, this would restrict the amount of oxygen
that is taken in from the germinating seeds.
In order to obtain further accurate results, many
different trials can be performed, where the
average of each trial can also to calculated. With
at least three trials, the data would be far more
accurate with each trial. We are unable to
accurately know which seeds are still
germinating, therefore, taking multiple trials
decreases error, which improves the accuracy of
the completed lab.

Bibliography:
1. Batty, David. "The Effect of Light on Germination and Seedlings." Effect of Light on
Germination and Seedlings. N.p., 1989 Web. 30 Oct. 2016. http://www.thompson-
morgan.com/effect-of-light
2. "Starting Seeds." Starting Seeds. N.p., n.d. Web. 30 Oct. 2016.
http://www.espsciencetime.org/student_life.cfm?subpage=326848

Station 4 Measuring the Rates of Photosynthesis

Table #6: Number of floating disks that rise in 0.5% bicarbonate solution and distilled water
solution, every minute under an artificial light.
 0.5% bicarbonate and dish soap Distilled water and dish soap

Minutes (min) Floating discs Total floating Floating discs Total floating
discs discs

0 0 0 0 0

1 0 0 9 9

2 0 0 0 9

3 0 0 0 9

4 0 0 0 9

5 0 0 1 10

6 0 0 0 10

7 1 1 0 10

8 4 5 0 10

9 5 10 0 10

10 0 10 0 10
A controlled variable is a method of controlling different aspects in a lab. The control is a
method of correcting information obtained in other experiments by removing all variable except
the variable that is looked for and valued.

Station 5 Separating pigments in Spinach Using Paper Chromatography

Table#7: The spinach transfer of pigment through paper chromatography shown through the
distance traveled from the original line that was created and the Rf calculations
 Spinach Colour Distance traveled from origin Rf calculations
(cm), (0.1cm)

Original spinach colour: 0.8 0.1

Chlorophyll A

Grass green: Chlorophyll B 2.1 0.31

Orange yellow: Carotene 2.8 0.41

Bright yellow: Xanthophyll 4.7 0.68

Sample Calculation:
Rf = Distance component traveled / Distance solvent traveled
Distance component traveled = 4.7cm
Distance solvent traveled = 6.9cm
= 4.7cm / 6.9cm
= 0.6811594203
= 0.68
% error
|(Experimental - theoretical) / (theoretical)| x 100%
Chlorophyll A: 0.1-0.33/0.33 x 100% = 69.7%
Chlorophyll B: 0.31-0.50/0.50 x 100% = 38.0%
Carotene: 0.41-0.58/0.58 x 100% = 29.3%
Xanthophyll: 0.68-0.92 x 100% = 26.1%
Safety
The ether solvent that was used in this experiment is highly flammable. This requires a
well ventilated area to perform the lab, and should not be inhaled. This can cause
respiratory paralysis and unconsciousness. This is also highly toxic.
During our lab, the complete rules for safety were followed with the fume hood, goggles
and aprons that we wore.

Bibliography:
1. "What Are the Side Effects of Ether Anesthesia?" LIVESTRONG.COM.
LIVESTRONG.COM, 2015. Web. 01 Nov. 2016.
http://www.livestrong.com/article/250029-what-are-the-side-effects-of-ether-
anesthesia/
2. "Chromatography of Simulated Plant Pigments." Chromatography of Simulated Plant
Pigments. N.p., n.d. Web. 01 Nov. 2016.
http://www.biologyjunction.com/chromatography_of_simulated_plan.htm
3. Ansell, Dave. "Chlorophyll Chromatography." The Naked Scientists. N.p., 21 Mar.
2010. Web. 01 Nov. 2016.
http://www.thenakedscientists.com/HTML/experiments/exp/chlorophyll-
chromatography/