Name of student: Marcelle Lamarche

Lab 2. Estimation of osmolarity in tissues by

bathing samples in hypotonic and hypertonic
solutions.
Total available points – 16

1. Background

Osmosis is a type of diffusion, the diffusion of water through a selectively permeable

membrane from a region where it is highly concentrated to a region where its concentration is
lower. The difference in concentration of water occurs if there is an unequal distribution of at least
one dissolved substance on either side of a membrane and the membrane is impermeable to that
substance. In this situation, the substance is called an osmotically active substance (OAS).
For example, if a membrane that is impermeable to sucrose separates a solution of sucrose from
distilled water, water will move from the distilled water where it is in a higher concentration,
through the membrane into the sucrose solution, where it is in a lower concentration. In this case,
sucrose is the osmotically active substance. (Biology, 2014)

Three terms, hypertonic, hypotonic, and isotonic, are used when referring to two
solutions separated by a selectively permeable membrane. The hypertonic solution has a greater
concentration of OAS than the solution on the other side of the membrane. It is described,
therefore, as having a greater osmolarity (solute concentration expressed as molarity). The
hypotonic solution has a lower concentration of OAS, or a lower osmolarity, than the solution on
the other side of the membrane. When the two solutions are in equilibrium, the concentration of
OAS is equal on both sides of the membrane, the osmolarities are equal and the substances are
said to be isotonic. The net flow of water is from the hypotonic to the hypertonic solution. When
the solutions are isotonic, there is no net flow of water across the membrane. (Biology, 2014)

The concept of osmotic pressure must be understood when studying osmosis. The
movement of water from a hypotonic solution through the membrane into a hypertonic solution
can be prevented by applying force or pressure on the hypertonic side. The force that must be
applied to prevent the osmotic movement of water from hypotonic to hypertonic, measured in
atmospheres, is referred to as osmotic pressure. Solutions with greater concentrations of OAS
have greater osmotic pressures because greater force is required to prevent water movement
into them. Distilled water has an osmotic pressure of zero. (Biology, 2014)

Research question:
What is the osmolarity of the potato (Solanum Tuberosum) cells, as measured by different
levels of osmotic pressure?

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 2. Variables
Variables
Independent variable
Solvent (H₂O).
Dependent variable
Organic potato tissue.
Controlled variables
1- Sample sizes.
2- Type of solute.
3- Type of solvent (H₂O).
Discuss the likely impact How the variable will be
on the investigation changed/ measured/
controlled
Using a solvent other than By using distilled water, the
water could have resulted in possibility of having
different particles invading undesired extra substances
the cells and performing or particles present in the
undesired reactions. experiment is reduced.
Potatoes are formed by plant These were cut into cubes of
cells, which have a 1 cm^2 that were stored in
semipermeable plasma sealed compartments to later
membrane surrounding them be weighted individually.
that enables osmosis to
happen.
Keeping sample sizes as Cutting each of the potato
similar as possible allows us cubes to even sizes to ensure
to measure each reaction it has little effect on the
with a closer degree of results.
accuracy than it would be
having vastly different sizes
across stations.
Salt is essential for osmosis Taking the solute in its purest
to happen. Using other form possible (Sodium
solutes, like sucrose, would Chloride, 99.9%) to avoid
trigger reactions within the unwanted particles.
cell irrelevant to the
experiment.
Making use of a substance By using distilled water rather
other than water would cause than tap water (or any other
the movement across the type for that matter) to avoid
membrane to be diffusion and unwanted particles or
not osmosis, defeating the contamination.
purpose of the experiment.
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 4- Air exposure. Leaving the potatoes out in
the air would kill and then
accelerate the decomposing
process of their cells,
rendering them useless for
the experiment.
Uncontrolled variables
1- Metal cutting The oxidation of the metal
materials. knives to cut the sample
could cause a consequential
reaction like oxidation of the
cells that could have
damaged some functions that
otherwise may impact the
reaction.
2- By hand cutting. It's not possible to cut
identical samples by hand,
which makes the collection of
data harder as it adds other
variables to the experiment.
3- External bacterial One of the testers (Arian)
infection. coughed on top of the
samples. His bodily bacteria
may have contaminated the
samples and could have
affected the cells’ reaction to
the solution.
4- Tint leakage. The pen and marker tint used
to label the samples most
likely stained the water and
cells with bacteria that may or
may not have affected the
testing.
All samples were placed in
sealed containers shortly
after being cut to delay
oxidation.
Using cutting machinery or
plastic materials properly
disinfected the sample’s
freshness could be better
preserved.
Using cutting machinery
allows precision and identical
sample sizes that minimize
variables.
By always using surgical
masks we could maintain a
sterile environment for the
research to happen.
Labelling the samples more
efficiently or re-organizing
them in a more specific sort
of way.
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 3. Materials

- Five 100ml beakers.

- One cutting board.
- Knife.
- Electronic scale.
- 2 large potatoes or 1/4 pumpkin or 3 beets. Vegetables must be uncooked and peeled.
- Ruler (+/- 1 mm)
- Timer.
-Tissue paper.
- Labels for containers and a marker.
- Water.
- NaCl (MW=58.44g/mol) concentrations: 0.2, 0.4, 0.6, 0.8 M solution, 100 ml each.

Table 1. Preparing the solutions for different molarities.

Molarity (CM), NaCl, grams (+/- 0.01 g) NaCl, teaspoons Water (up to), ml (+/- 0.00 ml)
M
0.2 1.17 0.25 100.00
0.4 2.34 0.5 100.00
0.6 3.51 0.75 100.00
0.8 4.68 1 100.00

3. Method

1. Label 5 containers as follows: 0.0M, 0.2M, 0.4M, 0.6M, and 0.8M.

2. Add 100 mL of the salt solutions to the correct container. (Note: water is 0.0M).

3. Use a sharp knife to cut twenty-five 1 cm cubes of the tissue of your choice. Use the ruler to
measure exact dimensions. It is very important that you have the same dimensions for all
twenty-five cubes.

4. Place potato cubes between the folds of a paper towel to blot sides and ends.

5. Weigh the cubes to the nearest 0.01 g on the aluminum sheet or paper towel on the balance.
Record the weight in Table 1 in the Results section.

6. Immediately transfer the vegetable cube to the water beaker (0.0 M).

7. Note what time the vegetable cubes are placed in the water beaker.

8. Repeat steps 5 to 7 with each cylinder, placing vegetable pieces in the appropriate incubating
solution from 0 to 0.8 M. Each solution must have 5 vegetable pieces.

9. Incubate for 1 hour. Clean up your workstation as you wait.

10. At the end of the incubation period, record the time when the pieces are removed in Table 1.

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11. Remove the potato pieces from the first sample. Blot the pieces on a paper towel, removing
excess solution.

12. Weigh the potato pieces and record the final weight in Table 2.

13. Repeat this procedure until all samples have been weighed in the chronological order in
which they were initially placed in the test solutions.

4. Ethical, Environmental, and Safety concerns

I consider unnecessary food waste to be unethical practice, hence waste was to be kept at a
minimum. Samples were evenly distributed and counted so there would be no excess litter.
Since our performing materials consisted of water, potato tissue, and sodium chloride, this was
safe to work with and dispose of without any special safety concerns.
The regular measures of lab robes, surgical masks, and the use of tweezers and other tools
when handling the samples were done accordingly, to avoid any alterations to the tissue,
solution or ourselves.

5. Results

Table 2. Times of incubation.

Time
Incubation start Incubation end incubated
Trial
time (+/- ____ s) time (+/- ___ s)
(+/- ___ s)

1 11:45 12:45 1:00

2 11:45 12:48 1:03
3 11:45 12:49 1:04
4 11:45 12:48 1:03
5 11:45 12:49 1:04
1 11:50 12:51 1:01
2 11:50 12:53 1:03
3 11:50 12:52 1:02
4 11:50 12:51 1:01
5 11:50 12:52 1:02
1 11:57 12:58 1:01
2 11:57 12:55 0:58
3 11:57 12:57 1:00
4 11:57 12:57 1:00
5 11:57 12:56 0:59
1 12:00 13:00 1:00
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2 12:00 13:02 1:02
3 12:00 13:01 1:01
4 12:00 13:03 1:03
5 12:00 13:01 1:01
1 12:03 13:05 1:02
2 12:03 13:04 1:01
3 12:03 13:05 1:02
4 12:03 13:06 1:03
5 12:03 13:07 1:04

Table 3. Changes in mass of potato tissue samples.

Initial Final
weight weight The
Weight
(g) (g) % Change in average Standard
Molarity Trial change
weight change in deviation
(+/- (+/- (g)
weight, %
g) g)
1 1.54 1.9 0.36 23.38%
2 1.2 1.5 0.3 25.00%
0.0 M
NaCl
3 1.07 1.37 0.3 28.04% 26.36% 0.05549775
4 1.29 1.71 0.42 32.56%
5 1.27 1.56 0.29 22.83%
1 1.37 1.53 0.16 11.68%
2 1.2 1.3 0.1 8.33%
0.2 M
NaCl
3 1.3 1.55 0.25 19.23% 11.00% 0.06906519
4 1.11 1.2 0.09 8.11%
5 1.18 1.27 0.09 7.63%
1 1.31 1.24 -0.07 -5.34%
2 1.07 1.06 -0.01 -0.93%
0.4 M
NaCl
3 1.26 1.18 -0.08 -6.35% -4.57% 0.03911521
4 1.32 1.3 -0.02 -1.52%
5 1.15 1.05 -0.1 -8.70%
1 1.2 1.08 -0.12 -10.00%
2 1.24 1.1 -0.14 -11.29%
0.6 M -
3 1.25 1.03 -0.22 -17.60% 0.05458938
NaCl 15.22%
4 1.21 0.96 -0.25 -20.66%
5 1.21 1.01 -0.2 -16.53%
0.8 M 1 1.3 1.04 -0.26 -20.0% -19.1% 0.03962323

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NaCl 2 1.23 1.07 -0.16 -13.01%
3 1.17 0.92 -0.25 -21.37%
4 1.01 0.78 -0.23 -22.77%
5 1.13 0.92 -0.21 -18.58%

To calculate the percentage change in weight, use this formula:

Percentage change in weight = weight change X 100

initial weight

If the sample gained weight, the value should be positive. If it lost weight, the value should be
negative.

To make calculations in Excel ---- highlight the values of all five trials for each set and use the
Excel formula “Ave(…)” to calculate the average. To calculate standard deviation using the same
set and formula “StDev(…)”.

Results from the experiment (averages) are to be analyzed in a graph format. It should be a
scatter plot with a line of best fit. Standard deviations should be shown as +/- values around the
averages (‘error bars’).

Graph 1. Average weight change and standard deviation.

Average Weight Change
40%

30%

20%
% Weight change

10%

0%
0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9
-10%

-20%

-30%

-40%
Molarity

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 6. Result from analysis
Analysis (6 grading points)
This criterion assesses the extent to which the student’s report provides evidence that the student has selected, recorded, processed
and interpreted the data in ways that are relevant to the research question and can support a conclusion.
Aspect 1: Raw Data
A1.1 Appropriate (insightful and thoughtful) qualitative data is recorded (observations).
A1.2 Appropriate quantitative data is recorded (or zero is awarded). (min of 5 increments and minimum of
A1.3 Raw data is clearly distinguished from processed data.
A1.4 Data is recorded correctly and honestly.
A1.5 Data is presented in a table including titles, units, and uncertainties.
Aspect 2: Data Processing
A2.1 Calculations to determine the DV are carried out, if necessary.
A2.2 Formulas (or excel equivalent) are stated before any worked example.
A2.3 Worked example for each calculation is shown.
A2.4 Calculations and statistics are appropriate for the investigation and RQ.
A2.5 Processed data and significant figures are consistent with the precision of raw data.
A2.6 Mathematics is correct.
A2.7 Processed data is presented in a graph.
Analysis

A2.8 Standard deviations are included where appropriate (minimum of 5 repeats needed to calculate the StDev)
A2.9 Statistical tests include full details: null and alternative hypothesis, degrees of freedom, critical values and probability levels.
Aspect 3: Impact of Uncertainty
A3.1 Uncertainties/errors are adjusted to reflect any calculations carried out.
A3.2 Error bars included, unless insignificant.
A3.3 Error bar source (e.g. standard deviation or min-max values) is stated and calculations are correct.
A3.4 Uncertainties are correct and explanations are given if necessary.
A3.5 Discussion of the magnitude of the uncertainties compared to the data collected. Ie. How do they affect the conclusion?
Aspect 4: Interpretation of Processed Data
A4.1 Patterns and trends in the graph or tables are described with reference to the graph/tables.
A4.2 Variation within the data (e.g. StDev) is discussed.
A4.3 Anomalies were identified and discussed.
A4.4 An appropriate choice of the graph is made.
A4.5 Patterns in the data discussed in relationship to the RQ.
A4.6 Data points are joined to illustrate the trend, or a best-fit line is drawn where appropriate. (Not applied if comparing two
means.)

 At what sodium chloride molarity does the curve cross the zero-change line on the
graph?

All samples have some degree of change in their mass. However, we can see how at the 0.4
NaCl Molarity the mass of the tissue begins to decrease (a ~-5% average between the samples
in that solution, look at Table 3) as osmosis happens, and the solutions begin to become
increasingly hypertonic.

 Explain how this information can be used to determine the osmolarity of the tissue.

To determine the osmolarity of the tissue, it is necessary we calculate the percentage of weight
change (as seen in Table 2) after the tissue is submerged into both hypertonic and hypotonic
solutions. The research is a necessary step because the data we recollected allows us to do the
calculation.

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  Estimate the osmolarity of the potato tuber tissue.
Based on the graph and the standard deviation of the 0.4 M NaCl solution, which was the closest
to being isotonic, we can roughly estimate the potato tissue’s osmolarity to be ~3.45 moles at the
point of zero change in the graph.

 What do your standard deviation values tell you about your sets of data?

The standard deviation in our results is constant and low, indicating small margins of error, and
great similarity in the overall reaction of the samples to the solutions. The data sets are clustered
around the mean, which represents a success in terms of accuracy.

 Were there any results from any trials that could be considered “outliers”? Why do you
think so?

No explicit outliers. The results represented in the graph indicate a specific trend of mass gain
and mass loss according to the increase in osmolarity. They are proving the hypothesis that if
osmolarity increases, then the mass of the tissue decreases because of osmosis and water loss
inside the tissue.

7. Conclusions

Evaluation (6 grading points)

This criterion assesses the extent to which the student’s report provides evidence of evaluation of the investigation and the results
concerning the research question and the accepted scientific context.
Aspect 1: Conclusion
Ev1.1 The relationship between the variables is correctly stated concerning graphs and tables.

Ev1.2 The possible impact of qualitative data is discussed.

Ev1.3 The extent to which data agrees with the RQ or hypothesis is stated and justified.
Ev1.4 The conclusion is based on information provided by the raw or processed data.
Aspect 2: Scientific Context
Ev2.1 The scientific explanation for the results is described.
Ev2.2 A scientific explanation is justified with explicit reference to data.
Ev2.3 A comparison with published data and theoretical texts, if possible.
Aspect 3: Limitations of the data and sources of error
Ev3.1 Reference to error bars concerning the validity of the conclusion. (e.g. Is the trend in results clear? Or the biological variation makes it
difficult to be certain of the trend? Refer to error bars, StDev, or correlation values)
Evaluation

Ev3.2 Analysis of the appropriateness of the range of the IV. (e.g. Was the range of values for IV sufficient to answer RQ?)
Ev3.3 Analysis of sufficiency of data. (e.g. Did the way the DV was measured produce valid results? Does it need to be changed in the future?)
Ev3.4 Anomalous points are identified and explained or stated if none.
All the following are evaluated in terms of possible effects on data and the magnitude of the error and could be presented as a
table.
Ev3.5 Systematic errors (problems with the method) are discussed. (e.g. identified controlled variables)
Ev3.6 Measurement/instrument errors (accuracy and precision).
Ev3.7 Any limitations to the experiment are discussed.
Aspect 4: Suggestions for Improvements and Extensions
Ev4.1 State suggestions for improvements to the methodology that are related to the RQ.
Ev4.2 Suggestions are specific (eg. Equipment named) and clearly explained.
Ev4.3 Improvements address all identified limitations and sources of error.
Ev4.4 All improvements are realistic and achievable. 9
Ev4.5 Improvements address the RQ quantitatively (improve control of IV, DV, or controls).
Ev4.6 Suggestions for a ‘scientifically interesting’ extension are based on the conclusion and are relevant to the RQ.
  Discuss how your results relate to your research question.

The research question dictated our purpose, to determine the osmolarity of potato tissue, and we
concluded in a close estimation thanks to the data recollected in the experiment. By submerging
potato tissue samples into both hypertonic and hypotonic solutions, we were able to corroborate
a trend in how vegetable tissue is affected by the osmolarity of its environment, and later offset
into an isotonic point (a result to the research question) estimated thanks to the record of mass
fluctuations caused by osmosis.

 What is the possible impact of your qualitative data?

Our qualitative data include slight changes in color, size, and overall appearance of the tissue
during the latter part of the experiment. It didn’t represent an impact on the results but was rather
an additional point of confirmation for our hypothesis. In the graph, we can see how the samples
with the lowest osmolarity increased considerably in weight. Something visible in great contrast
when observed side by side with the tissues that were exposed to the highest osmolarity levels,
since these samples were lesser than what we had begun with.

 Are there any articles published that relate to your findings?

In one article I found, the relation of temperature and tissue with osmosis was debated,
questioning whether they had an impact on water movement or the separation of solvent and
solute from the solution. We used room temperature solutions while the author experimented with
different temperatures and tissues, like leaves and animal cells. It relates to our findings, and
supports us to an extent, by agreeing on the necessity water has for balance (isotonic
environments) and that being the reason osmosis happens in the first place.

8. Evaluation

 What was successful in your experiment?

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We were successful in responding to the research question by estimating, graphing, and
visualizing the aftermath of osmosis and changes to plant tissue osmolarity.

 What are the potential improvements for your experiment?

First, I believe there could be an improvement in the equipment used, especially the scale. The
weight ins were fluctuating constantly because of external causes unrelated to the scale itself;
like pressure on either side of the working table, for example. A newer scale would help increase
accuracy in this and future experiments.

Secondly, the sealing. I suppose we could’ve preserved the potatoes better during the weigh-in
process. Keep them fresher. The ones in the 0.0 M NaCl are perhaps better maintained than
those in the 0.8 M NaCl solution, as they were introduced to the solution first and didn’t wait in
the air.

Finally, time calculation. We calculated time until minutes, and I think our error bars (Graph 1)
and standard deviation could have been reduced even more by augmenting specificity in the
data.

The potential improvements for this experiment are slight. We worked neatly and with a decent,
functional structure that helped us get through the experiment successfully.

 Suggest a ‘scientifically interesting’ extension for the experiments you performed,

based on the conclusion you made.
It would be interesting to see what effect temperature has on osmosis by having solutions with
variated temperatures, measured by a thermostat, with the aid burners to maintain it constant. It’s
commonly said that, when cooking, it’s better to dissolve the water at room temperature. Because
it allows the solution to have adequate levels of salinity and for the tissue (potatoes especially) to
better absorb flavor. We could evaluate that if solutes dissolve better at lower temperatures, then
it would be easier to maintain an isotonic environment at higher temperatures. Or even just try to
confirm if said belief is accurate.

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 9. Bibliography
“Biology: Course Companion.” Biology: Course Companion, by Andrew Allott and David
Mindorff, Oxford University Press, 2014, pp. 2–8.

Hammel, H. T. (1999). Evolving Ideas about osmosis and capillary fluid exchange. Faseb
Online Library. Retrieved September 10, 2022.

Communication (4 grading points)

This criterion assesses whether the investigation is presented and reported in a way that supports effective communication of the focus,
process, and outcomes.
Aspect 2: Structure
C2.1 An obvious clear structure was used (titles, headings, etc).
C2.2 Graphs, tables, and figures are clearly labeled and titled (e.g. Figure 1……..).
C2.3 Graphs, tables, and images are included as close as possible to their first use/reference. All images, graphs, and tables are referred to in
the text (e.g. Fig1).
C2.4 Tables and graphs do not break across pages.
C2.5 Graphs are clear with appropriate coloring.
Aspect 3: Focus
C3.1 All data and graphs are relevant to the RQ.
C3.2 All images and media are relevant to the RQ.
C3.3 All citations included are relevant to the RQ
C3.4 All background information and discussion are relevant to the RQ.
Communication

C3.5 The report is easy to read and ‘flows.

C3.6 The report is 6-12 pages.
Aspect 4: Terminology and Conventions
C4.1 Tables are labeled correctly including titles, units, and uncertainties (±….).
C4.2 Table column headers are correct (IV in the first column, DV values in the subsequent columns).
C4.3 Significant digits are consistent with the precision of the equipment throughout the report.
C4.4 Processed data has decimal places consistent with calculations.
C4.5 Raw data is clearly distinguished from processed data.
C4.6 An appropriate graph representation of processed data was chosen.
C4.7 Graph axes are labeled clearly, including titles, SI units, and uncertainties.
C4.8 Graph axes are scaled appropriately.
C4.9 All images are annotated.
C4.10 Mastery of and fluency in the use of scientific terminology is demonstrated.
C4.11 The use of excessive jargon and words is avoided.
C4.12 Non-standard technical terms are explained and used correctly.

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