ACS Chemistry Laboratory Supplement

Project-Based Labs

Kidney Stones
Introduction
The analytical laboratory of a major research hospital has employed your team. You have
been asked to investigate the formation of kidney stones. In many areas of the country, kidney
stones are a severe problem and their treatment is not well understood. Your task is to
investigate the causes of these stones and design ways in which the formation of the stones might
be prevented in the body. This may allow you to recommend treatments for patients suffering
from these stones.
Kidney stones are formed when ions react to form an insoluble precipitate. The stones are
composed of an inorganic salt, sometimes complexed in an organic matrix. You will be provided
with soluble sources for many of the most common ions in the body, including cations of
sodium, potassium, calcium, and magnesium. You will also have soluble sources for common
anions in the body, including chloride, oxalate, phosphate, hydrogen phosphate, and dihydrogen
phosphate.

Project Tasks
Some of these tasks will be accomplished experimentally and some by library or web-based
research in appropriate resources.
1. Determine what ions are present in kidney stones.
2. Replicate the formation of kidney stones in the lab.
3. Predict which combinations of ions will produce kidney stones in the lab.
4. Find patterns in the solubility of compounds formed.
5. Understand why some mixtures of ions do not seem to react while others result in
a precipitate.
6. Find ways to use the properties of these ions to prevent formation of precipitates.
7. Manipulate conditions to make precipitates more soluble.
8. Comment on which conditions used in the lab might be compatible with physiological
processes.
9. Determine if the size of a lab kidney stone affects its solubility.

Learning and Performance Objectives

1. Understand the principles of solubility of ionic compounds and be able to predict the
solubility of a given compound from its formula.
2. Understand the differences in reactivity between large stones and smaller ones.
3. Suggest ways to redissolve a kidney stone under laboratory conditions.
4. Suggest ways to prevent the formation of kidney stones, at least under laboratory
conditions.
5. Understand the effect of pH on formation and solubility of kidney stones under
laboratory conditions.
6. Understand how complexing agents are useful in increasing solubility.

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7. Use your chemical knowledge to solve a real world problem.

8. Handle solutions of acids and bases safely.

Resources Available
1. A strategy for determining solubility patterns
2. A general approach for preparing kidney stones of larger size
3. Two different approaches for redissolving a prepared kidney stone

Reports
1. Planning sheets are to be completed as directed by your instructor.
2. The final report for this project should follow the basic guidelines given previously.
Be sure to include:
a. observations and other results from all group experiments.
b. your interpretation of which compounds commonly form kidney stones
and why they do so.
c. differences in appearance and results for freshly prepared kidney stones,
large crystals, and smaller crystals.
d. results from the solubilization procedures.
e. results from precipitation prevention procedures.
f. possible suggestions for patients suffering from kidney stones, based on
your experimentation.
3. Be sure you support your decisions with data from the various aspects of the project,
including cost, safety, environmental impact, and cleaning effectiveness.

Materials Available: Chemicals

Soluble salts: NaCl, CaCl2, KCl, MgCl2, Na2C2O4, Na2CO3, NaH2PO4, K2HPO4, Na3PO4
0.1 M HCl, 1 M HCl, 6 M HCl, 0.05 M EDTA
Safety Information
You may encounter some or all of these materials during your experimentation for this project.
Hydrochloric acid is corrosive and causes severe burns if concentrated. Notify your instructor if
any acid is spilled. Flush the affected area with running water for 15 minutes if you should spill
any on your skin. Be sure to read the label carefully and use the concentration called for.
Sodium hydroxide is corrosive and causes severe burns if concentrated. Notify your instructor if
any base is spilled. Flush the affected area with running water for 15 minutes if you should spill
any on your skin.

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When mixing acids and bases use extreme care mix drop wise with plenty of stirring. If the
solution gets hot, stop mixing for a while and let the solution cool.
When mixing concentrated acid and water, always add the acid to the water and not the other
way around. Gentle stirring while adding the acid is also recommended.

Techniques you may need to study

Drying and weighing compounds
Growing crystals
Measuring pH
Qualitative solubility tests
Small-scale chemistry techniques
Titration
Vacuum filtration

Resource: Finding Solubility Patterns

The first task is to determine if there are any solubility
patterns in the salts that you have available. This information
will help you find which combinations of anions and cations
are candidates for the formation of kidney stones. You will be
given soluble sources of the anions and cations that are
prevalent in the body. These include compounds containing
Na+, K+, Ca2+, Mg2+, Cl, PO43, HPO42, H2PO4, and C2O42.
Small-scale techniques can help you perform these tests
qualitatively and in an organized fashion. The 24-cellwellplate
and plastic Beral-type pipet shown in the photograph are convenient for your initial small-scale
investigations. Toothpicks can be used to stir the materials in each cell. You may want to place a
piece of white or colored paper under the well plate to enable you to see fine precipitates. If you
have any difficulty drawing conclusions about solubility, you can move to macroscale
techniques. Larger amounts of each salt can be dissolved in a larger amount of water in a test
tube before mixing in a 50-mL beaker. Although the macroscale approach may be somewhat
slower and consumer more materials, it may be needed to check questionable observations.
Resource: Preparation of a Sample Kidney Stone Compound
Once you have identified a potential kidney stone compound, plan to prepare about 1 gram of
the compound. You will need to calculate how much of each starting material you will need, and
then prepare aqueous solutions of each. Mixing the solutions should give you the required
material. This material will need to be separated and dried. Knowing the expected yield is 1.0 g,
calculate the actual percent yield of the salt you obtain.
Many kidney stones grow quite large (several grams is not uncommon) and therefore you
will need to prepare the stones by several different methods to enable you to look at very fine
powder and larger crystals. Often larger crystals can be grown by leaving a seed crystal in a
solution of the salt for longer period of time (a week for example).

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Resource: Strategies for Solubilizing Kidney Stone Compounds

There are at least two possible approaches to solubilization of the kidney stones, both of
which involve the removal of one of the components of the stones by application of a competing
process. The first is to remove the anion using its acid-base character and the second is to remove
the cation by complexing it with a chelating agent.
Strategy 1. Use the acid-base character of the anion. Oxalate and phosphate are the anions of
weak acids, it should be possible to manipulate the hydrogen ion concentration so that the anion
joins with a hydrogen ion (becomes protonated) and is removed from the solubility equilibrium.
Given a salt MA, where M is a cation and A represents an anion, these are the equilibrium
expressions.
Equation 1

MA(s) M+(aq) + A(aq)

Equation 2 A(aq) + H+(aq) HA(aq)

If HA is a weak acid, the equilibrium position in Equation 2 will be shifted to the right. Le
Chteliers Principle tells us that the equilibrium position in Equation 1 will also shift to the right
to produce more anion, bringing more of the kidney stone compound into solution.
Equation 2 also suggests that it might be possible to solubilize kidney stones by changing the
pH of the solution. Look up the Ksp for each potential kidney stone compound you have
identified. Use the Ksp to calculate the amount of anion in solution. This will be a very small
amount for any highly insoluble kidney stone compound. Then use the Ka of the conjugate acid
of that anion to calculate the hydrogen ion concentration that will be in equilibrium with that
amount of anion. From that value you can calculate the pH, which should be the pH at which the
kidney stones will dissolve.
You should investigate the effect of changing the pH on a number of differently prepared
samples. One plan is to treat the process like a titration. The kidney stones should be in an
Erlenmeyer flask as a slurry in water, the acid can be placed in a burette and added drop by drop
as you swirl the slurry in the Erlenmeyer flask. The pH can be monitored with a pH meter or
with the use of appropriate indicators. Do you think that the size of the crystals might affect the
solubilization process? Predict and then experiment to verify or revise your prediction.
Controlling the pH may also be helpful in preventing the formation of kidney stones, another
goal for this project lab. What effect do you think changing the pH will have on the formation of
the precipitate? Predict and then experiment with the synthesis of kidney stone compounds under
different pH conditions. Are you predictions confirmed by your observations?
Strategy 2. Remove the Cation by Using a Chelating Agent.
Another way to shift the solubility equilibrium to the side of the dissolved species is to
remove the cation instead of the anion. EDTA stands for ethylenediaminetetraacetic acid. It is
capable of holding cations very strongly. Such materials are called chelating agents. Using
EDTA (or its sodium salt) makes it possible to remove some cations from the solubility
equilibrium. Given a salt MA, where M is a cation and A represents an anion, these are the
equilibrium expressions.

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Equation 3

Project-Based Labs

MA(s) M+(aq) + A(aq)

Equation 4 M+(aq) + EDTA(aq) M(EDTA) complex(aq)

If the EDTA complex tightly binds the metal ion, the equilibrium position in Equation 4 will
be shifted to the right. Le Chteliers Principle tells us that the equilibrium position in Equation 3
will also shift to the right to produce more anion, bringing more of the kidney stone compound
into solution.
EDTA is not very effective with metal ions with a charge of +1, but forms a stable one-toone complex with Ca2+. Knowing the concentration of EDTA, you can calculate the volume of
EDTA required to solubilize a known mass of kidney stones. Investigate the effect of EDTA on
solubilizing kidney stones, using an experimental procedure similar to the previous acid process.
Use a slurry of the kidney stones in the Erlenmeyer flask and a solution of EDTA in the buret.
How will you determine the endpoint in these titrations?
Adding a chelating agent may also be helpful in preventing the formation of kidney stones,
another goal for this project lab. What effect do you think the presence of EDTA will have on the
formation of the precipitate? Predict and then experiment with the synthesis of kidney stone
compounds in the presence of EDTA. Are you predictions confirmed by your observations?

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Kidney Stones: Planning Sheet 1

To be completed before starting experimentation.

Group Name

State the overall purposes of this project in your own words.

1. What ions are present in the compounds you will be using?

2.

Are all these ions present in the human body? Which are the most abundant?

3. Where in the body would you find the most abundant ions?

4. Your first task is to prepare a solution of each compound. The exact concentration is not
important for preliminary testing. Outline the process you will use to prepare a small amount
of solution for quick tests.

5. When you have prepared solutions of all the compounds provided, your task is to identify
which combinations of ions will result in the precipitation of an insoluble salt. Plan to use a
24-cell well plate as shown here. Indicate how you will arrange the compounds to obtain the
maximum amount of information with the minimum number of tests.

A
B
C
D

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Kidney Stones: Reflection and Planning Sheet 2
To be completed after the first laboratory period.

Project-Based Labs
Group Name___________________

State the purposes of this weeks lab in your own words.

1. Which combinations of ions produced an insoluble product? Write a balanced molecular

equation and then a net ionic equation for each reaction that produced a precipitate. Be sure
to indicate which species are in aqueous solution and which are solids.

2. Which combinations of ions produced only soluble products? How would their equations
differ from those in question 1?

3. What are the formulas of the insoluble compounds you produced? How do you know that
these are the insoluble compounds?

4. What general rules can you come up with to predict whether an ionic compound will be
soluble in water? Can you see any patterns in your results?

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5. Next lab you will produce samples of all the insoluble compounds produced. Each person in
your group should produce at least one insoluble salt. Assume you need to produce 0.5 g of
product. Give a sample calculation here of how much of each starting material you plan to
use. Indicate which reagent you will use as the limiting reactant.

6. In order to separate your solid from the solution, you will need to filter it using vacuum
filtration. Use the Techniques section and other resources available to you to review this
method of separation. Sketch and label the appropriate apparatus and briefly outline the
procedure in your own words.

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Kidney Stones: Reflection and Planning Sheet 3

Group Name___________________
To be completed after the second laboratory period. Before you leave lab, prepare one more
batch of each of your insoluble salts. This time, leave the solution until next week to separate the
product from the solution. Next week, you will compare the products formed during lab today
with those that have formed over the intervening week.
State the purposes of this weeks lab in your own words.

1. Indicate the appearance of the product, the amount of starting materials, and the theoretical
yield for each product made by your group. Note you will not be able to record the actual
mass of product until next week when it has dried. If additional rows of data are needed, use
the product preparation records on the back of this sheet.
Product Preparation Records
Formula
of
product

Appearance
of product

Mass
Reagent 1

Moles
Reagent 1

Mass
Reagent 2

Moles
Reagent 2

Theoretical
moles of
product

Theoretical
mass of
product.

2. Describe how you separated the solid from the solution. Did you have any problems with this
procedure? How did you overcome them?

3. Do the results of your experiments support the known composition of kidney stones that you
found by web or library research? Why or why not?

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4. Next week we will begin to look at ways to get rid of kidney stones. Remember that even
though your compounds appear to be insoluble, there are always some small amounts of the
compounds cation and anion in solution. To help you begin to think about this, write an
equilibrium expression for what happens when a small amount of one of your insoluble salts
dissolves in water.

5. Write out the reaction(s) that would occur if you added acid to one of your insoluble salts.
Explain why the addition of acid can result in the solubilization of your compound.

6. Write out the reaction(s) that would occur if you added EDTA to one of your insoluble salts.
Explain why the addition of EDTA can result in the solubilization of your compound.

7. Look up and record the Ksp of your salt and the Ka of the conjugate acid of the salts anion.
Give a reference for the source of each value. You will need this information for next weeks
calculations.

Product Preparation Records, contd.

Formula
of
product

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Appearance
of product

Mass
Reagent 1

Moles
Reagent 1

Mass
Reagent 2

Moles
Reagent 2

Theoretical
moles of
product

Theoretical
mass of
product.

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Kidney Stones: Reflection and Planning Sheet 4

To be completed after the third laboratory period.

Group Name___________________

State the purposes of this weeks lab in your own words.

1. Indicate the theoretical yield, experimental yield, and percent yield for each compound you
made last week.
Product

Theoretical Yield

Experimental Yield

Percent Yield

2. Is there a difference in the percent yield for each compound? What factors can lead to a
lower than 100% yield? Are there any factors that might produce a higher than 100% yield?

3. Record observations on your preliminary solubilization experiments. Try the solubilization

process on freshly prepared salt (salt you made today), a sample of the dried, filtered salt (salt
made last week), and a sample of the large crystal salt (salt left over the week in the reaction
beaker).
Formula of
Compound

Kidney Stones

Observation on addition of acid

Fresh
Filtered
Large
crystal

Observation on addition of EDTA

Fresh
Filtered
Large
crystal

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4. How did the solubilization of the dried, filtered solid compare with the larger crystals formed
over the week, and the newly prepared material still in solution? If there was a difference,
what factors do you think might account for the difference?

5. Summarize the results of your experiments in preventing precipitate formation.

Formula of
Compound

Observations on synthesis in
presence of acid

Observations on synthesis in
presence of EDTA

6. Next week you will use todays results to do quantitative solubilization experiments. You
will use measured amounts of kidney stones and known concentrations of acids and bases.
Use the Ksp of your salt and the Ka of the anions corresponding weak acid to calculate the
approximate pH at which you expect your salt would be soluble. Show your work clearly.

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Kidney Stones: Reflection and Summary Sheet 5
To be completed after the fourth laboratory period.

Project-Based Labs
Group Name___________________

State the purposes of this weeks lab in your own words.

1. Briefly summarize the different procedures that you have used to prepare and study kidney
stones.

2. Did the pH at which your kidney stones dissolved correlate with the pH that you predicted?
Explain why or why not.

3. Did the amount of EDTA required to solubilize the kidney stone correlate with what you
predicted? Explain why or why not.

4. Based on your experimentation, what possible recommendations do you have for a person
who suffers from kidney stones?

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Kidney Stones