0Lab: Investigating the importance of buffers in living systems

Even small changes in pH can cause distress to organisms. The changes in hydrogen ion concentration in cells
can change the rates of some chemical reactions. For example, the pH of human blood is normally about 7.4
and must be in the range of 7.0-7.8 for survival. If the pH is in the 7.0-7.3 range the person will feel tired, have
trouble breathing, and may even be disoriented. If the pH of the blood is in the 7.5-7.8 range, the person will
feel dizzy and rather agitated.
It is very important for organisms to be able to maintain a fairly constant internal environment (homeostasis).
To prevent the hydrogen ion concentration of the cytoplasm from changing too much, cells have chemical
compounds called "buffers" that will bind with hydrogen ions when their concentration increases too much.
Buffers can also release bound hydrogen ions when their concentration in the solution decreases too much.

In this laboratory, students will measure the pH of 2 common acid and base solutions, as well as how several
materials respond to the addition of an acid and a base to determine whether living materials have buffering
capacity.

Essential Question
How do buffers regulate pH in living systems?

Materials
pH paper 50 ml beakers buffer solution dixie cup (trash)
various homogenates (potato and yeast) 0.1 M NaOH 0.1 M HCl
pipettes

Procedure - Day 1
1. Using a small piece of pH paper (tear into small pieces) and forceps, find the pH of the HCl and NaOH. Use
the pH scale noted on the side of the pH paper container.
2. Fill in Data Table 1. Place used pH paper in the trash Dixie cup.
3. Fill one 50 ml beaker (sample beaker) with 10 ml of buffer solution using the 10ml cylinder found with the
solution.
4. Split your group into two teams.
5. Team 1: Using a small piece of pH paper and forceps, find the pH of the buffer solution. Add 10 drops of
HCl. Find the pH again. Add 10 more drops. Find the pH again. Add 10 more drops. Find the pH again.
Rinse your beaker. Repeat using water then each of the homogenates. Record your data in Table 2.
Team 2: Using a small piece of pH paper and forceps, find the pH of the buffer solution. Add 10 drops of
NaOH. Find the pH again. Add 10 more drops. Find the pH again. Add 10 more drops. Find the pH again.
Rinse your beaker. Repeat using water then each of the homogenates. Record your data in Table 3.
6. Exchange data with the other team. Fill in the appropriate Data Table.
7. Transfer your initial and final pH from Data Tables 2 & 3 into Table 4.
8. Clean up your lab area as directed.

Procedure - Day 2
1. The teacher will demonstrate how to create a digital graph of your data using google sheets.

Data Table 1
Substance pH Substance pH
0.1 M HCl 1 0.1 M NaOH 13

Data Table 2
Substances pH with 0 drops pH after 10 drops pH after 20 drops pH after 30 drops
HCl (initial) HCl HCl HCl (final pH)
Buffer + HCl 7 8 8 8
Water + HCl 7 4 1 1
yeast + HCl 7 6 4 2
Potato + HCl 7 6 5 4

Data Table 3
Substances pH with 0 drops pH after 10 drops pH after 20 drops pH after 30 drops
NaOH (initial) NaOH NaOH NaOH (final pH)
Buffer + NaOH 7 7 8 9
Water + NaOH 7 12 8 8
yeast + NaOH 6 7 8 9
Potato + 6 10 10 17
NaOH

Data Table 4
Substance Initial pH Final pH Change in pH
(final pH - initial pH)
Buffer + HCl 7 8 1
Water + HCl 7 1 6
yeast + HCl 7 2 5
Potato + HCl 7 4 3
Buffer + NaOH 7 9 2
Water + NaOH 7 7 0
yeast + NaOH 6 9 3
Potato + NaOH 6 17 11

Analysis and Questions:

A negative control is a control group in an experiment that uses a treatment that is not expected to produce
results. A positive control is a control group in an experiment that uses a treatment that is known to produce
results.
1. Which of the above is the negative control?
water
2. Which is the positive control?
buffer
3. Graph your results from data tables 2 and 3 in a line graph in the space below. Use google sheets to
complete the graph. Label lines or provide a key.
Graph 1 -
 Graph 2 -

NOTE:
You will be expected to make a graph digitally as the teacher explains how to correctly construct it.
 4. Fill in the draft CER form below.

Title of Inquiry/Lab: Investigating the importance of buffers in living systems

The purpose of this form is to guide you in the process of creating a Claim-Evidence-Reasoning (CER)
argument. Complete the following portions in order. Because this is a draft, you may use bullet
points/phrases/notes.

The guiding question is what the inquiry is designed to answer.

Guiding Question:
How do buffers regulate pH in living systems?

Evidence should include enough to support the claim. The evidence used should be relevant. When possible it
should include qualitative (using 5 senses), and quantitative (numerical) data. On your final draft, include a
data table to organize your information. If the information is numerical, insert an appropriate graph.
Possible: (Data table, rough draft graph & observations)
- Data table 2 and 3
- Line graphs 2 and 3
- pH paper color changes
- buffers are weak buffer and changes that can react with weaker acids
- Acids have high ph values.

Now for the analysis:

● List the important pieces of evidence from above and how they lead to a supported answer to the
question (also known as the claim) For each key piece of evidence, you will need some reasoning.
The reasoning should explain how and why that piece of evidence shows support. It should also
include the relevant supporting science concepts or principles. Look for information from the
background section of the lab and your notebook. Do not use quotes, but use in-text citations for
information from outside sources. Add rows to the chart below as needed. This chart will be used to
write the reasoning section of the final draft CER. The purpose of the table below is to organize your
information to make it easier to write a strong reasoning section. You must use these key pieces of
evidence in your reasoning section to support your claim.

Evidence
→ Reasoning (cite if from outside source)

→
Buffers stabilize ph amount This shows that buffers stabilize ph levels.
In data table 2 the ph amount stays more
consistent when the buffer is applied

→
The line graph shows the line moving in This shows the buffer keeping the line consistent.
a more consistent fashion.

→
The ph color changes. Shows the effect of the acids on a physical substance.

→
Acids have a strong ph value Shows the adverse effect acids would have on the body
without proper mitigation.
 →
Bases have a low ph value Show the lack of a negative effect that buffers have.

Based upon your evidence and reasoning, state a claim that is supported. It should be one sentence. It
should describe the relationship between the dependent and independent variables. It answers the question,
“What can you conclude?” (In this case, what can you conclude about how buffers regulate pH in living
systems?), and does not include a simple “Yes or No”.
Claim: Buffers serve to keep the ph value of acids in the body stable so they do not hurt the body.

From this working draft, you will write your final CER. It will either be in essay form: Claim, Evidence,
Reasoning with a data table and graph embedded in the evidence section, or it will be on a whiteboard to be
presented to the class. Whiteboards are often arranged as follows:

Guiding Question:

Claim:

Evidence Reasoning

Please fill out the whiteboard form below: It will be the final draft that the
teacher grades for accuracy.
Remember: Your claim is an answer to the science question you are trying to solve. The evidence is the data
you collect and analyze that will be used to explain your claim. The reasoning is the explanation of your claim
using your data for support and the science to explain why.. The science information is what we have learned
in class that will help you explain your data.

Title:
Guiding Question: What is the role of buffers in
homeostasis?
Claim: Buffers keep the ph level of acid-stable to prevent
harm to the body.
Evidence Reasoning
When the buffer is added the
pH level stabilizes and does
not fluctuate dramatically
when more acid is produced
as it would otherwise. This
can be seen in the data table
as the line with the buffer
changes less.