Operation Water Biology

Lesson One

Chlorination and Dechlorination

The purpose of a water treatment plant is to take raw water from a well or
fresh water source, remove all of the contaminants and make the water safe to
drink. The contaminants that should be removed include bacteria, ammonia,
phosphorus, nitrogen, dissolved organic material, iron, manganese, arsenic and
More information on Drinking
many more. Health Canada is a government organization which sets guidelines on Water Quality Guidelines can be
the maximum level of different contaminants that can be present in the treated found at
www.safewater.org/fact-
drinking water. As long as the treatment plant takes out enough contaminants to sheets-1/2017/1/23/
meet these guidelines, the water is assumed to be safe to drink. There are purposeguidelinesregulations
guidelines for levels of iron, arsenic and nitrate because these things may have
direct aesthetic or health consequences, as we will discuss later. In Canada, there
are no guidelines for ammonia or phosphorus because it is not usually dangerous to
drink water with naturally occurring levels of these chemicals. The guidelines were
set by looking only at the direct effect each contaminant could have on someone
drinking the water. We now have more information about the way that some
contaminants interact with each other and other consequences of their presence. If
you look at the whole picture it seems that the contaminants that do not have
guidelines should have them and the guidelines that are already in place are not
strict enough. The experiments and lessons in this kit will use iron, ammonia and
chlorine as examples to explain why this is a problem and show how it can be
solved.

In this lesson we will begin talking about chlorine. When most people think
about chlorine the first thing that comes to mind is the smell of swimming pools.
Although you probably cannot smell the chlorine in your tap water it is added to
most municipal drinking water supplies as part of the treatment process.

Much like in a swimming pool, chlorine is added to tap water to disinfect it.
This is usually done as the last stage of water treatment after the water has been
filtered in other ways. Chlorine can kill microorganisms like bacteria and viruses
which could cause a wide range of diseases and health problems. Chlorine also More information on
reacts with, and removes, dissolved chemicals such as ammonia. Some of the chlorination can be found at
chlorine gets used up every time it removes a contaminant from the water and www.safewater.org/fact-
sheets-1/2017/1/23/what-is-
these reactions produce small amounts of other chemicals called chlorine chlorination
byproducts. These byproducts can sometimes be dangerous if they build up in large
amounts. This can happen if a lot of chlorine is added to water that has not been
filtered well and still has a lot of contaminants in it.

www.safewater.org Lesson One Handout Page 1

 Chlorine is useful to treat water because it reacts with so many different
things and killing microorganisms is very important but if you want to study
chemical reactions in water the chlorine there can sometimes get in the way. In a
future lesson we will want to use some water that does not have any chlorine in it
so today we will begin the process of taking the chlorine out of some tap water.
This process is called dechlorination.

This experiment will show how granular activated carbon (GAC) can remove
gases such as chlorine from water. The GAC has a very large surface area and is full
of tiny holes. This allows it to grab and hold gases in the same way that a sponge
holds liquids.

Materials needed for this experiment are:

One total chlorine test strip

Plastic cup
2mL of Granular Activated Carbon (GAC)
Empty 5mL vial
Empty, well rinsed, 2L plastic bottle
Watch or timer
Tape and a marker

1. To know how much chlorine gets taken out of your water by the GAC you
will need to know how much is in it at the beginning. To find this, fill a cup
about halfway with tap water then follow the instructions in the total Total chlorine
chlorine concentration test procedure from the test procedure sheet. Be concentration of tap water
sure to keep the cup. You can rinse it out and use it again in the next few
experiments.
2. Using the 5mL vial, measure out 2mL of GAC. When pouring GAC from one
container to another you should do it over a clean piece of white paper so
that any you spill can be poured back into its original container.
3. Put your 2mL of GAC into the empty 2L plastic bottle and then fill it up with
tap water and put on the cap. The 5mL vial needs to be rinsed out right
away to get the dust from the GAC out of it. This vial will be used in future
lessons so it must be clean.
4. Swirl the water around so that the GAC spreads throughout the water. The
GAC will settle to the bottom but you want it to be exposed to as much of
the water as possible so hold the bottle upside down for a few seconds so
that the GAC falls through the water then turn it right side up so it falls
through again.
5. Label your bottle so it does not get mixed up with those of other groups.
6. You will open the bottle and test the chlorine concentration again in a few
days after the GAC has had enough time to absorb the chlorine.

www.safewater.org Lesson One Handout Page 2

 Operation Water Biology

Lesson Two

Chlorine

Since chlorine gets used up when removing contaminants from water, the
more contaminants there are the more chlorine is needed to get rid of them. The
total amount of chlorine that must be added to water to fully disinfect it is known
as that water’s chlorine demand. For good sources of water the chlorine demand is
low and the water treatment facility only has to add a small amount of chlorine to
disinfect the water and make it safe to drink. Some extra chlorine is usually added
to make sure that no bacteria can grow in the pipes and distribution system and More information on
that the water is still clean when it comes out of your tap. In fact, unlike most of the chlorination can be found at
www.safewater.org/fact-
other water quality guidelines which state the maximum recommended amount of sheets-1/2017/1/23/what-
a contaminant in your drinking water, the guideline for chlorine is the minimum is-chlorination
allowed amount. Any time that all of the chlorine gets used up before the water
comes out of the tap there is a risk that bacteria could begin to grow in the water
pipes. The current guideline for total chlorine from the Guidelines for Canadian
Drink Water Quality is a minimum concentration of 0.5mg/L. Did your tap water
meet this requirement when you tested it in the previous lesson?

It is easy to show the effect that a contaminant can have on the total
chlorine concentration of a sample of water. In this experiment you will see that
the chlorine demand created by a little bit of tea is enough to use up all of the
chlorine in a water sample.

Materials needed for this experiment are:

Two total chlorine test strips

Two plastic cups
Tea bag
Empty 5mL vial
Watch or timer

1. Use the 5mL vial to collect 5mL of the chlorine solution that your teacher
has prepared.

2. Pour this sample into a cup then fill that cup about 3/4 of the way up with
tap water to dilute the solution. Swirl the water around a little bit to make
sure it is well mixed.

www.safewater.org Lesson Two Handout Page 1

 3. Fill the second cup about 1/4 of the way up with this diluted chlorine Initial total chlorine
solution. The first cup should still be about half full; set it aside for now and concentration of chlorine
solution
perform a total chlorine concentration test on the sample in the second
cup.

4. Once you have the result from that test you should empty and rinse the
second cup.

5. Dip the tea bag into the first cup two or three times until the water just
begins to turn colour. Do not let the tea bag sit in the water. If the water
changes colour too much it could affect the colour of the test strip when
you do the test. The tea bag can be thrown away or composted. Total chlorine
concentration of solution
6. Do a total chlorine concentration test on the sample that you dipped the after tea bag exposure
tea bag into. When you have the results from the test all the water can be
disposed of. The cups should be rinsed and kept.

www.safewater.org Lesson Two Handout Page 2

 Operation Water Biology

Lesson Three

Ammonia and Chloramine Part One

One of the most important chemicals to the water treatment process is

ammonia. The chemical formula for ammonia is NH3. Ammonia is something that
many water treatment facilities deal with in one way or another. It is common for
More information on the
ground and surface water sources to contain ammonia because ammonia can come
nitrogen cycle can be found
from so many sources. Ammonia can be added to soil by nitrogen-fixing bacteria as at http://
part of the nitrogen cycle, decay of plants and animals or agricultural and industrial chemistry.about.com/ od/
geochemistry/ss/
processes. Ammonia is highly soluble so it gets dissolved and transported by nitrogencycle.htm
surrounding ground water.

In the areas that do have ammonia in their raw water it is a very

problematic source of chlorine demand. For each milligram of ammonia in the
water it takes 10-15 mg of chlorine to react with it and get rid of it. The reaction
between ammonia and chlorine is much faster than the rate that chlorine kills
bacteria so you cannot use chlorine to disinfect water that contains ammonia.
Unfortunately the most widely used method of removing ammonia is to add
chlorine. In a process called "break-point chlorination" chlorine is continuously
added to water until all of the ammonia and bacteria have been removed, or in
other words, until the chlorine demand has been met.

This works if there is only a little bit of ammonia but if there is more than More information on
0.3 mg/L ammonia in the raw water then so much chlorine would have to be added disinfection byproducts can be
found at
to get rid of it that it would result in dangerous levels of chlorination byproducts. http://www.lenntech.com/
You can see that there are cases where the only options seem to be, a) not using processes/disinfection/
byproducts/disinfection-
break-point chlorination and thus leaving bacteria in the water, or b) disinfecting
byproducts.htm
the water at the risk of adding harmful amounts of chemicals to it. This means that
some treatment facilities have to use very complicated and expensive methods,
which often still involve the use of other chemicals, to take ammonia out of the
water before they add chlorine.

www.safewater.org Lesson Three Handout Page 1

 One new option that communities with ammonia problems have is
biological filtration. This is a safe, chemical free, method of removing ammonia. In
a biological filtration facility one of the stages of filtration is to pass the water
through a special filter that is full of nitrifying bacteria. These bacteria take in the
ammonia and some oxygen and perform a bio-oxidation reaction. They oxidize the
More information on biological
ammonia into nitrite, NH3 + O2 -> NO2- + 3H+. Then further oxidize that into nitrate, filtration facilities can be found
NO2- + H2O -> NO3- + 2H+. The bacteria gain energy from these reactions and are at
specialized to do them very efficiently. This process is part of the natural nitrogen www.safedrinkingwaterteam.o
rg/ibrom.html
cycle and does not produce any harmful byproducts. The nitrate that is produced by
this process can easily be removed from the water by the reverse osmosis
membrane in the final stage of the filtration process.

The reaction between chlorine and ammonia can be written as

NH3 +HOCl -> NH2Cl + H2O. In this chemical equation NH3 is ammonia and HOCl is
hypochlorous acid which is formed when the chlorine first dissolves in the water.
The primary result of this chemical reaction is NH2Cl, a chemical know as
chloramine. Chloramine is a disinfectant like chlorine, it is a weaker disinfectant
than chlorine but it lasts much longer in water. The chlorine concentration in water
can gradually decrease as the chlorine evaporates out but chloramine does not do
this. This makes it useful for making sure water stays disinfected throughout
drinking water distribution systems. In areas where there is no, or very little,
ammonia in the raw water treatment facilities might still want to use chloramine for
this purpose. After chlorinating (disinfecting) the water, as the last step in the
treatment process they add ammonia and more chlorine to the water so that they
react and create chloramine.

With this information, you may be wondering if there is chloramine in your

own tap water and how you might be able to measure it. The fact of the matter is,
part of the process of finding chloramine concentration is the total chlorine
concentration test that you have already done. The other part is a second kind of
test which is called a free chlorine test. Understanding the difference between free
and total chlorine is very important. The free chlorine test finds the concentration Example
If you test a water sample for
of regular unreacted chlorine like the kind added to water during the chlorination
both free and total chlorine and
process or the kind found in chlorine bleach. The total chlorine concentration test get values of 2.0 mg/L free
finds the combined concentrations of the regular unreacted chlorine and the chlorine and 2.5 mg/L total
chlorine then you know that the
chlorine that has been in a reaction and is now chloramine. Since total chlorine is concentration of chloramine in
free chlorine plus chloramine, the total chlorine concentration must always be that sample must be 0.5 mg/L.
greater than or equal to the free chlorine concentration. This also means that the
total and free chlorine tests can be used together to find the chloramine
concentration of a water sample. Subtracting the free chlorine concentration from
the total chlorine concentration will give you the chloramine concentration.

www.safewater.org Lesson Three Handout Page 2

 Consider an experiment that could put this information to use. Imagine two
beakers, each with exactly 100mL of water. The first has a free chlorine
concentration of 20mg/L, a total chlorine concentration of 20mg/L and an ammonia
concentration of 0mg/L. The second beaker has both free and total chlorine
concentrations of 0mg/L and an ammonia concentration of 4mg/L. What do you
think would happen if the contents of these two beakers were poured together into
a larger beaker?

This problem can be broken into smaller pieces that should be considered one at a
time.

1. What is the chloramine concentration in each of the two original beakers?

2. Pouring the beakers together will result in a total volume of 200mL; this
volume change could dilute the chemicals. Might this have an effect on the
concentrations of chlorine and ammonia before any chemical reaction even
occurs? Consider the result of two 100mL beakers being poured together if
one had 20mg/L of salt and the other had 4mg/L of sugar.

3. Assuming that a free chlorine concentration of 10mg/L is exactly enough to

react with 1mg/L ammonia, what should the final free chlorine and
ammonia concentrations be?

4. If all of the free chlorine used in the reaction becomes chloramine what
should the final chloramine and total chlorine concentrations be?

www.safewater.org Lesson Three Handout Page 3

 Operation Water Biology

Lesson Four

Ammonia and Chloramine Part Two

In this experiment you will combine an ammonia solution with a chlorine

solution in the same way that was described in Lesson 3. You will test the two
samples separately to find their individual ammonia, free chlorine and total
chlorine concentrations. 100mL of each will then be poured together to see if a
reaction resulting in the production of chloramine really takes place.

To make sure that there are no extra, unwanted reactions taking place and
to control as many variables as possible, the solutions will be mixed using some of
your dechlorinated water.
Materials needed for this experiment are:
Four total chlorine test strips and three free chlorine test strips
Three ammonia test strips
Ammonia colour matching card/chart and square test vial or
disposable beaker if you have the new version of the ammonia
test.
Two plastic cups
Two 250mL beakers
Empty 5mL vial
5mL ammonia solution vial
Coffee filter
Dechlorination bottle prepared in Lesson One
Watch or timer
Tape and marker
Stir stick

First you can find out how much chlorine the GAC was able to remove from your
tap water.

1. You will be using a coffee filter to separate the GAC out of the water. The Figure 1 Figure 2
easiest way to do this is by folding the filter into a cone. Begin by folding
the filter in half and then half again so it is in the shape shown in Figure 1 to
the right. Looking at the filter from above, you should see four edges of the
filter like in Figure 2. You can pull three of these edges in one direction and
the forth in the other direction to open the filter into a cone shape.

2. Hold the coffee filter in place over a plastic cup. Open the dechlorination
bottle and slowly pour some of the water through the filter until the cup is
about half full. This filter will be used again in step 5.

www.safewater.org Lesson Four Handout Page 1

 3. Perform a total chlorine concentration test on this sample. Total Chlorine
concentration of tap water
4. Find the difference between the chlorine concentrations in the regular tap after exposure to GAC
water that you tested in the first lesson and this water that has been
exposed to the GAC.

Now begins the ammonia / chloramine experiment

5. Fold your coffee filter into a cone and hold it in place over one of the Amount that the GAC
beakers. lowered the tap water's
total chlorine concentration
6. Slowly pour water from your dechlorination bottle through the coffee filter
until the beaker is full to the 250mL mark. Move the filter to the second
beaker and fill it to 250mL as well.

7. Label one beaker as "Ammonia" and the other as "Chlorine".

8. Pour the contents of the 5mL ammonia vial into the ammonia beaker.

9. Swirl the beaker so that the ammonia solution will be well mixed. If you
want to use an object to stir the solution use a clean stir stick, not a pen,
pencil, finger or anything else.

10. Perform the Ammonia Test on this sample by following the provided
procedure. You must wait 5 minutes before reading the result of the All results from these tests
Ammonia Test so go on to step 11, the time should be up when you are should be recorded in the
finished the other tests. space provided on page 4.

11. Pour about 50mL of your diluted ammonia solution into a cup and perform
a total chlorine concentration test. Empty and rinse the cup, pour another
50mL of your diluted ammonia solution sample into the same cup and do
the Free Chlorine Test. Empty and rinse the cup again.

12. Use the 5mL vial (not the one that the ammonia came in) to get 5mL of the
chlorine solution that has been prepared by the teacher.

13. Pour this sample into the chlorine beaker and swirl it to mix the solution. If
you want to use an object to stir the solution use a clean stir stick, not a
pen, pencil, finger or anything else.

14. Do all three tests on this chlorine sample in the same way you did for the
ammonia sample. Use the second cup for the chlorine tests of this sample.

15. Pour some of the water out of the ammonia beaker until you have 100mL
left in the beaker. You may want to pour the water into a cup so that if you
pour too much you can put some back.

www.safewater.org Lesson Four Handout Page 2

 16. Pour some of the water out of the chlorine beaker until you have 100mL
left. You may want to pour it into the other cup so that if you pour too
much you can put some back.
All results from these tests
17. Pour your 100mL chlorine solution into the ammonia beaker so that there should be recorded in the
is 200mL in this beaker. Swirl this solution a bit to mix it, if you want to use
space provided on page 4.
an object to stir the solution use a clean stir stick, not a pen, pencil, finger
or anything else.

18. Do all three of the tests on this mixed sample. Either of the cups can be
used for the chlorine tests.

1. When you have all nine of your test results calculate the chloramine
concentrations of each of the three solutions tested. Remember that you
can do this by subtracting the free chlorine concentration from the total
chlorine concentration. Record these values in the spaces provided.

2. Calculate the concentrations of all four chemicals that would have been in
the mixed solution after pouring together the chlorine and ammonia
solutions but before the chemical reaction started (just like the salt and
sugar example). The differences between these calculated values and the
actual concentrations you found experimentally are due to the chemical
reaction.
3.
a. Did the chemical reaction appear to change the concentration of all
four of the chemicals you tested for?
b. Did any of your calculated concentrations from question 2 match,
or almost match, the actual experimental results? If so, does this
mean that these things were not involved in the chemical reaction?
How might you explain this?
c. Were any of your calculated concentrations from question 2 very
different from the actual experimental results? What does this
mean?

4. In the Lesson 3 questions we made two assumptions. First we assumed that

a free chlorine concentration of 10mg/L is exactly enough to react with
1mg/L ammonia and secondly we assumed that all of the free chlorine used
in the reaction becomes chloramine. Do you think your experimental
results confirm these assumptions? Why or why not?

www.safewater.org Lesson Four Handout Page 3

 Lesson Four Results Sheet

Ammonia Solution Chlorine Solution Mixed Solution

Ammonia Concentration Ammonia Concentration Ammonia Concentration

Total Chlorine Concentration Total Chlorine Concentration Total Chlorine Concentration

Free Chlorine Concentration Free Chlorine Concentration Free Chlorine Concentration

Chloramine Concentration Chloramine Concentration Chloramine Concentration

www.safewater.org Lesson Four Handout Page 4

 Operation Water Biology

Lesson Five

Consequences of Inadequate Drinking Water Treatment

Discovering if a community has a problem with ammonia is as simple as

doing a single test on a sample of tap water but since there is no guideline for Location that this water
sample was collected
ammonia in the Guidelines for Canadian Drinking Water Quality this test is rarely
done. People in these communities often do not know that their water is not being
properly disinfected. In this lesson your class will perform ammonia tests on the
water samples that you have collected.
Ammonia concentration of
the water sample
Your class might find ammonia in some of the water samples that you have
collected and tested during this lesson. Recall from Lesson Three when you learned
that some water treatment facilities add ammonia to the water. This is done after
the water has been disinfected so that they can use it to create chloramine which
will keep the water disinfected throughout the distribution system. In these cases
the water is properly disinfected even though an ammonia test might show that
there is still ammonia in the water. This practice is usually only used by water
treatment facilities serving very large distribution systems and is not common in
rural communities that have small facilities. If you do find ammonia in these
samples you should investigate whether the facility that treated that water added
the ammonia after disinfection or if the ammonia was in the raw water. Only if the More information on drinking
water advisories in Canada can
ammonia was in the raw water is there cause to be concerned that the water was
be found at
not disinfected. This means that there could be disease-causing micro-organisms in https://www.ec.gc.ca/
that water. When a situation like this is discovered a boil water advisory is usually indicateurs-indicators/
default.asp?
put in effect. lang=en&n=2C75C17A-1

www.safewater.org Lesson Five Handout Page 1

 Operation Water Biology

Lesson Six

Removing Iron from Drinking Water

Iron is an essential element for humans. People usually get all the iron they
need from the food they eat so extra iron in water is not necessary for good health.
The guideline for iron in the Guidelines for Canadian Drinking Water Quality is
0.3mg/L. This guideline is in place primarily for aesthetic reasons since iron
discolours water and promotes bacterial growth. It is not a health-based guideline
because iron, and the kinds of bacteria that grow around iron, are not usually
harmful to peoples’ health. These iron bacteria do cause many other problems
though. When iron is present in water, bacteria that use iron as their energy source
will grow and build up anywhere that water flows. These bacteria form thick layers
called biofilm in reservoirs and pipes, causing a lot of damage to distribution
systems that is difficult and expensive to repair. Excessive bacterial activity also
results in water with an unpleasant taste and smell which makes it bad for both
cooking and washing. These bacteria can still grow in water that has less iron than
the 0.3mg/L guideline. If there is any iron at all that these bacteria can use for
energy then they will grow and spread. To keep iron bacteria out of drinking water
all of the iron must be removed, not just enough to meet the guideline.

Take a look at the tubes of raw well water that are included in the kit. You
will see that the iron in this water has settled to the bottom of the tubes, you
should see many very small orange particles there. If you shake up one of the tubes
and look very closely you can see the tiny orange particles floating all around in the
water. These particles are visible because this iron is insoluble in water. This is not
always the case though, when this water first comes out of the well there would
not be any tiny orange particles visible in it. This was because of a difference
between the two chemical states of iron. Iron can either be in a reduced state or an
oxidized state (also known as rust). One of the biggest deciding factors of which
state iron is in is exposure to oxygen. When the iron is deep in a well it is not
exposed to oxygen so it will be in its reduced state. This reduced iron is soluble in
water so it dissolves and you can not see it even though it is still there. When the
water is brought up from the well and is exposed to oxygen it becomes oxidized.
This is when the iron separates from the water because it is no longer soluble.

www.safewater.org Lesson Six Handout Page 1

 Making sure that all of the iron in the water is oxidized is an important step
in filtering it out. This is because when the iron is still reduced it is dissolved in the
water and can pass right through filters along with the water. To filter out the iron it
must first be oxidized so that the particles separate from the water and then can be
picked up by the filter. There are a few different methods that treatment facilities
use to oxidize iron and other contaminants. The simplest is to let the water sit in a
large open pool called a reservoir so that it is exposed to the oxygen in the air, most
of the iron will eventually oxidize and settle to the bottom but this process is slow
and often leaves some reduced iron in the water. Another option is to use oxidizing
chemicals; these can be added to the water to oxidize all of the iron more quickly
and efficiently than the reservoirs. These processes create water like the raw well
water that you saw in the tubes with many very small iron particles floating around.
Even though the iron is no longer dissolved in the water it is still very difficult to
filter out because the particles are so small. This iron can pass right through most
regular filters and when very fine filters that can catch particles of that size are used
they clog up very quickly and need to be constantly cleaned and replaced. The usual
solution to this is to add another chemical called a flocculent. The flocculent makes
the little particles stick together in clumps which are called floc. This floc is easier to
filter out because its larger size makes it easier to catch.

A different option for filtering iron is to use a biological process like the one
for ammonia that was discussed earlier. Rather than putting chemicals in it, the
water can be passed through a filter containing bacteria that do an even better job
of oxidizing all of the iron. These are actually the very same iron bacteria as we More information on biological
mentioned at the beginning of this handout, we may want to keep them out of the water treatment can be found
at
water pipes but we can still put them to work for us in our filters. Iron bacteria take www.safedrinkingwaterteam.o
in reduced iron and oxygen and perform a bio-oxidization reaction on them which rg/ibrom.html
produces oxidized iron. These iron bacteria gain energy from performing this
reaction and they are specialized to do it very effectively. There is a second benefit
to using these bacteria as well, as they are oxidizing the iron they are also making a
little bit of sticky gel which automatically forms a floc with the iron. This allows the
bacteria to do the job of both the oxidizing and flocculating chemicals at once.
During the experiment for this lesson you will see an iron floc formed by iron
bacteria in the other tubes included in the kit. These bio-oxidized water samples
were taken from real biological filters in a water treatment facility where iron
bacteria oxidized the iron and formed the floc. Once this floc is formed it gets
caught in the filter and the water passes through iron free. There is no longer an
energy source in this water for any other iron bacteria to use so they can not grow
and form biofilm and thus all of the problems associated with iron in water are
avoided.

www.safewater.org Lesson Six Handout Page 2

In this experiment you will be able to see many of the things discussed in this lesson
for yourself. You will filter water samples that have been oxidized in different ways
and see if they give different results.

Materials needed for this experiment are:

Three small plastic beakers

Two 250mL beakers
Plastic cup
Raw well water sample
Bio-oxidized water sample `
Two coffee filters
Four aluminum packets of iron test reagent
Marker
Scissors
Stir stick

1. Label one of the small plastic beakers “raw unfiltered” then shake up the
raw well water sample, open it and pour 10mL into that beaker. Set this
beaker aside for now.
2. Fold your coffee filter into a cone just like in Lesson Four and hold it in
place over the cup.
3. Slowly pour the rest of the raw well water sample through the filter into
the cup.
4. Label one of the small plastic beakers “raw filtered” then pour 10mL of
filtered raw well water from the cup into that beaker. This beaker should
also be set aside for now. Any water still in the cup can be disposed of.
5. Put 100mL of tap water into one of the large beakers.
6. Very gently and slowly turn the tube of bio-oxidized sample upside down. If
the floc is stuck to the bottom of the tube turn it over a few times until all
of the floc is knocked loose, you want all of it to come out when you pour
the water out. You must do this carefully; you do not want to break the floc
apart too much.
7. Open the tube of bio-oxidized sample and very carefully pour it into the
beaker with 100mL of water (not through a filter). Try to hold the tube
close to the surface of the water when pouring because the floc might
break apart when it hits the water if it falls too far.
8. Fold your second coffee filter into a cone and hold it in place over the
second large beaker.
9. Very slowly pour the sample you just mixed in step 7 through the coffee
filter. Try to hold the beaker close to the filter when pouring so that the
water does not fall as far.
10. Label the third small plastic beaker “filtered bio-oxidized” and pour 10mL of
the filtered bio-oxidized sample into that beaker.

www.safewater.org Lesson Six Handout Page 3

Once everyone has prepared their three samples the teacher will prepare the
unfiltered bio-oxidized samples and give one to your group.

11. You can now begin observing and comparing the different samples.

The pairs of samples that you should compare are:

a. Unfiltered raw water to unfiltered bio-oxidized
b. Unfiltered raw water to filtered raw water
c. Unfiltered bio-oxidized to filtered bio-oxidized
d. Filtered raw water to filtered bio-oxidized

On a piece of paper write down the names of the two samples you are
comparing and then write down your observations in a sentence or two.
Leave lots of space to add things later then put the names of the next pair
of samples and continue.

Examine them closely, does there seem to be differences between them?

Are there different sizes or number of particles visible? Does one look like it
has more or less iron than the other?

The aluminum packets contain a reagent that is used to indicate the presence of
iron. When it is added to a water sample it will turn pinkish red if there is iron in it.
The darker the colour gets the more iron there is. This is very useful for comparing
samples, it can be put into a number of samples and you will know that the one
that turns the darkest shade of red has the most iron or if some turn the same
colour you will know that they have the same iron concentration.

12. With scissors very carefully cut open the tops of your four packets and pour
one of the packets into each of your four samples. Use a stir stick to mix the
powder into the water samples. You must wait at least three minutes for
the reaction to finish but once the colour changes there is no hurry, the
samples will stay that colour for a long time.

13. When the three minutes are up compare the same four pairs of samples
again. Which sample really had more iron? Judging by how different the
colours of the samples are does the difference in iron concentration seem
to be large or small? Write down your observations in a sentence or two in
the spaces that you left under your first observations.

www.safewater.org Lesson Six Handout Page 4