Quantitative Inorganic Analysis Laboratory

Formal Report
Date Performed: 20
October 2015
Date Submitted: 30
October 2015
Institution/Department:
Department of
Chemical Engineering
College: Engineering
Instructor: Cris Angelo
Pagtalunan

Quantitative Determination of Dissolved Oxygen

Content by Winkler Redox Titration
M. K. Coo
Dissolved oxygen (DO) content is an essential tool in determining the water
quality and ability to sustain life. The experiment was done to determine the DO
content and to evaluate its ability to sustain life and its level of pollution. Winkler
method was used to determine the DO content of a water sample from a pond near
Vinzons Hall. In this method, Mn(III) becomes Mn(II) once acidifies (with the
presence of I -). Given the reaction 2MnO + 6H + + 2I- 2Mn 2+ + I- + 4H2O, the I2
is approximately equivalent to the DO content which was titrated with the sodium
thiosulfate solution. The DO content was calculated from the volume readings.
The experiment yielded 3.25 ppm O2. From this DO content, it can be concluded
that the pond is highly polluted and cannot sustain life for larger organisms such
as fishes. Possible sources of error include volatilization of iodine, exposure of
the sample to air while the Mn 2+ and I- solutions were being added and the
presence of the bubbles in the sample (faulty in sampling).

Quantitative Determination of Dissolved Oxygen Content by Winkler Redox Titration

Coo, 2015| 1

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Formal Report
Introduction

Clean, good water

89

Dissolved oxygen (DO) content is essential to

the water quality of an aquatic system because of its
capability to determine or estimate life sustainability. It
also indicates the presence of pollutants in water.
Organic waste discharges from cooling towers directly
affect dissolved oxygen content in water. Organic
materials are broken down by microorganisms. This
process requires oxygen and as a result, decreases
dissolved oxygen content.1 Because of this, it can be a
good indirect indicator of the health of aquatic systems
since high levels of pollution or organic material will
increase the oxygen consumption of the system. This is
important because if the total amount of dissolved
oxygen is lower than what is ideal, aquatic life may
become stressed
Because of this, can be a good indirect indicator
of the health of aquatic systems since high levels of
pollution or organic material will increase the oxygen
consumption of the system. This is important because if
the total amount of dissolved oxygen is lower than what
is ideal, aquatic life may become stressed, i.e. exhibit
erratic behavior, move away, or die. Generally, warmwater fish require less oxygen than cold-water fish. 2 The
dissolved oxygen levels needed by aquatic life is shown
in Table 1 and the equivalent pollution levels in Table 2.

DO content is affected by water temperature,

salinity, atmospheric pressure and altitude, suspended
matter, and aquatic plants and animals. Aquatic plants
increase DO in water through photosynthesis. This
requires light and so, DO also varies with the clarity of
the water, the time of day, and the seasons.1 The location
also affects DO content. In stagnant systems such as
lakes and ponds, the DO content varies with depth and
in river systems, DO content varies along its length.
Locations with high turbulence tend to have more
dissolved oxygen than stagnant locations. 3
One method used to measure dissolved oxygen
content is the Winkler method. In the Winkler method,
manganese (II) is added to the sample. This reacts with
hydroxide ions to form Mn(OH )2 (Equation 1)
which reacts with the dissolved oxygen as shown in
Mn(OH )3
Equation 2, producing the brown
precipitate.1 Manganese (II) sulfate was added before KI
because the oxidation-reduction reaction between

and O 2 is too slow. 5

I
Mn(OH )2
[1]
2++2 OH
Mn
[2] 4 Mn (OH )2 +O 2+ 2 H 2 O 4 Mn (OH )3

If the system does not contain oxygen, the

Table 1. Dissolved oxygen and aquatic life

2+

Mn

addition of
Dissolved
oxygen

Pollution degree

causes the white

Mn(OH )2

precipitate to form. 1 By acidifying the solution,

manganese (III) hydroxide dissolves and reacts with

iodide ( I ) to form iodine ( I 2 ). (See Equation

Cannot sustain life
Can sustain life for 3). Iodine then forms a complex with excess iodide to

warm-water fish
form I . This is shown by Equation 4. Iodine is
7-11 ppm
Ideal for cold-water fish
3
volatile so exposure to air can reduce its amount.

Table 2. Dissolved oxygen and pollution degree

is less volatile and its formation improves
I
0-2 ppm
4.5-6.6 ppm

Dissolved
oxygen (ppm)

below 4.5
4.56.6
6.77.9

Pollution degree

Highly polluted
Moderately polluted
Slightly polluted

Chem 31.1 Section, Room No., Time

accuracy. 1

2++ I 2+ 6 H 2 O
+ 2 Mn
[3]
+ 2 Mn (OH )3 +6 H
2I

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+
I 2 I 3
[4]
I

The amount of oxygen in the system is directly

I3

related to the amount of

of

I 3

produced. The titration

with thiosulfate can then be done to

determine DO content in the sample. The reaction is

shown by Equation 5. 1

2+3 I

[5] S 4 O 6
2+ I 3

2 S2 O 3

The goal of this experiment was to measure the DO

content and assess the ability of the system to sustain
life and its level of pollution.
Materials and Methods
The Winkler method is an iodometric method.
Iodometry is a titration that involves the use of iodide as
a reducing agent. In contrast, iodimetry is a titration that
uses iodine as an oxidizing agent. One limitation of
iodimetry is that iodine is a weak oxidizing agent so
some reactions, especially those with weak reducing
agents, may not be complete.4,5
Approximately 0.15 g of KI O 3 was dissolved in
50 mL of distilled water in a beaker. The solution was
then quantitatively transferred into a 100-mL volumetric
flask and diluted to mark. Three 10-mL aliquots were
then taken and placed into 3 separate Erlenmeyer flasks.
Twenty milliliters of water , about 1 gram of KI and 10
mL of 0.5 M H 2 SO 4 were then added to each flask.
Excess potassium iodide was added to the KI O 3
to convert it to

I 3

as shown in Equation 4 and 6.

The volatility of the iodine produced in Equation 6 is

reduced by its reaction with excess iodide. This makes
the iodine more stable and helps reduce the loss due to
volatility.6
[4] I- + I_2 [I3]-

+ 3 I 2+ 3 H 2 O
+ 6 H
[6]
+5 I
I O 3

Chem 31.1 Section, Room No., Time

This reaction must be done in acidic conditions.

Sulfuric acid was added because Equation 6 will not
proceed under neutral or basic conditions. KI was added
first in order for the reaction to proceed to completion. 7
After addition of these reagents, the solution was
immediately titrated with Na 2 S2 O 3 solution until it
was pale yellow. Starch was then added. This caused the
solution to turn dark blue and the titration was
continued until the blue color disappeared. This was
done with the solution in each Erlenmeyer flask.
Starch was added as an indicator in this experiment.
It forms a blue complex with

I3

and the colors

disappearance marks the endpoint of titration. 7 This

reagent must be added near end of titration because the
complex dissociates slowly when it was formed in the
presence of numerous

I3

. The endpoint may be

delayed because the color change will not be as sudden.

Also, starch breaks down in acidic conditions so this
affects the indication of endpoint which is why the
boiling is maintained all throughout.4
A glass bottle was covered with aluminum foil for
sample collection. This was conducted at a pond in
Vinzons Hall. The bottle was submerged in the water
and tilted at around 45, and was capped underwater
only when there were no more bubbles coming from the
bottle. Capping underwater prevents air from getting in.
This can change the DO content of the sample. Taking
care not to produce air bubbles in the solution, 0.5 mL
of 4.0 M Mn SO 4 was added before adding 0.5 mL
of the NaOH-KI- Na N 3 . The bottle was then closed,
taking care not to include air bubbles, and then shook
thoroughly. The cover was removed and concentrated
phosphoric acid was added. The bottle was covered and
shaken. A 50-mL sample was taken from the bottle and
placed in an Erlenmeyer flask. The solution was then
Na 2 S2 O3 solution until pale
titrated with the
yellow. Starch was then added and caused the solution
to turn blue and the titration was continued until the
color disappeared. This was done three times.
The titration of the iodine with thiosulfate is shown
by Equation 5.

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2+3 I

[5] S 4 O 6
2+ I 3

2 S2 O 3

Given these values, the concentration of dissolved

oxygen can be calculated using the stoichiometry of the
Winkler titration. Also, using Tables 1 and 2, the
pollution level and ability to sustain life of the system
can be assessed. These are shown on Table 4.
Table 4. Measured Dissolved Oxygen Content (ppm)

The stoichiometry of the standardization

procedure is one mole of iodate is equivalent to 6 moles
of thiosulfate and in the Winkler titration, four moles of
thiosulfate is equivalent to one mole of oxygen.
Results and Discussion
In this experiment, one of the aims was to
measure the DO content using the Winkler method. The
sample was mixed with manganese (II) sulfate and a
solution of sodium hydroxide, potassium iodide, and
sodium azide ( Na N 3 ). The resulting solution was
then acidified with phosphoric acid and titrated with
sodium thiosulfate.
In the procedure, the manganese (II) sulfate was
added before the NaOH-KI- Na N 3 solution because
the overall reaction that forms the iodine will be faster
this way. The reaction between iodide and oxygen is
slow and is hampered by the presence of NaOH, which
was added to promote the production of Mn(OH )2 ,
as this process prefers a low pH.
The NaOH-KI- Na N 3 must be added immediately
after the manganese (II) sulfate because of
Mn(OH )3
disproportionation.
The
can
disproportionate into

2+
Mn

and

MnO2 . This

makes the concentration of Mn(OH )3 decrease so

less of that species will be available for the formation of
iodine. This decreases the calculated DO concentration. 1
Table 3 shows the volumes of thiosulfate used per
trial.
Table 3. Measured volumes for Winkler titration

Volume
sample
Volume
titrant

TRIAL 1

TRIAL 2

TRIAL 3

of

50.0 mL

50.0 mL

50.0 mL

of

1.6 mL

1.6 mL

1.5 mL

Chem 31.1 Section, Room No., Time

DO content
Average DO
content
Aquatic life
Pollution level

TRIAL 1

TRIAL 2

TRIAL 3

3.32

3.32
3.25

3.12

Cannot sustain life for large

organisms i.e. fishes
Highly polluted

Waste water usually contains contaminants

other than organic material. One of these contaminants
is nitrogen in the form of nitrite. This species is toxic to
aquatic life and is usually oxidized into nitrate. 9 In cases
where the nitrate concentrations are high, nitrite may
still remain in the water. This interferes with the titration
by reacting with iodine in a redox reaction. This would
decrease the volume of thiosulfate used and would
decrease the calculated DO. Sodium azide was added to
react with the nitrite to form N 2 and N 2 O . 1
Another type of contaminant is metals. Copper and
iron can interfere with this method. The presence of
copper catalyzes the oxidation of thiosulfate with air.
This would increase the volume of titrant used, and
would increase the calculated DO content. 7 Presence of
iron also interferes with the Winkler method and is a
possible source of error. Other contaminants may also
interfere with the titration reaction by reacting with
oxygen, iodine or iodide. 8
The presence of microorganisms and organic
material in the sample also causes error. After
collection, these microorganisms continue to decompose
the organic material, consuming oxygen in the process.
This decreases the calculated DO content. It is
impossible to obtain a sample from the environment
without the presence of these so one way to lessen the
effect is to immediately add Mn SO 4 and NaOH-KINa N 3 . The inclusion of fish and other organisms
that consume oxygen in the sample also causes the same
type of error. However, the inclusion of plants or algae
will cause the DO content to increase through
photosynthesis. This was prevented by covering the

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bottle in foil to keep the sample in darkness. Plants
absorb oxygen in order to break down the carbohydrates
produced during photosynthesis and maintain their
metabolism. However, when photosynthesis cannot
occur, plants absorb more oxygen than they release and
this causes the DO content to decrease. 9 If the sample
was left to stand overnight, the organisms that are in the
sample would decrease the DO content.
Under acidic conditions, excess iodide can be
oxidized by oxygen in the air, forming additional

I 3 . This is why the titration has to be carried out

immediately, to minimize error resulting from this. This

error increases the concentration of thiosulfate in the
standardization and increases the calculated DO
content.8
Another potential source of error is the presence of
dissolved carbon dioxide in the thiosulfate solution.
This promotes the disproportionation of thiosulfate into
bisulfate and sulfur. This reduces the concentration of
the titrant over time and can increase the volume used
for the titration. One way to help prevent this is to boil
the distilled water before use. 7 Bacteria can also cause
decomposition in solutions that were not freshly
prepared. A way to counteract this is to use fresh
solutions, boil the water that will be used in preparing
the solution or add chloroform to a bottle of thiosulfate.
Light also catalyzes the decomposition so it is best to
keep light exposure to a minimum.
In the measurement of DO content, the location of
the sample origin is also very important. In rivers and
streams, the turbulence, vegetation, and current of the
river changes along its length and these factors affect
DO content.4 In order to have an accurate picture of DO
content, the samples must be taken from different
locations around the river.
Another source of error would be the sampling.
In our sample obtained, the bottle had bubbles present in
it indicating the presence of air. This would result to an
increase in DO content since air contains more oxygen,
thus making it a positive error.7

2+
, oxygen, and
Mn

I . The DO
content calculated was averaging 3.32 ppm O 2 .
the reaction of

This oxygen concentration indicates high level of

pollution and that the stream cannot support large
water organism life. The experiment goals have been
met since the DO content has been identified and the
degree of water pollution and life-sustaining ability of
the stream has been assessed.
To improve accuracy, it is recommended that the
experiment be repeated multiple times, taking samples
from different parts of the stream to have a more
accurate picture of the health of the aquatic system.
Also, it is recommended that the time between sample
collection and sample analysis be reduced to avoid DO
changes caused by microorganisms and contaminants.
Make sure to follow correct procedure in sample
collection to reduce errors from air bubble production. It
is also important to mind the order of addition of
reagents because any change from the procedure can
cause errors and that the titration of iodine must be done
quickly to reduce errors from iodine volatility.

References
[1] Ibanez, J.G., Hernandez-Esparza, M., Doria-Serrano,
C., Fregoso-Infante, A., Singh, M.M. Environmental
Chemistry; Springer Science: New York, 2008.
[2] Swedish Environmental Protection Agency.
Environment Quality Criteria: Lakes and Watercourses.
http://www.naturvardsverket.se/Documents
/publikationer/620-6032-5.pdf (accessed October 26,
2015).
[3] United States Geological Survey. Water properties:
Dissolved
Oxygen.
http://water.usgs.gov/edu/dissolvedoxygen.html
(accessed October 26, 2015).

Conclusions and Recommendations

[4] Christian, D.G. Analytical Chemistry, 6th ed.; John

Wiley & Sons: Hoboken, 2004.

The goal of the experiment was to determine

DO content of a water sample and using that value,
assess the water quality and the aquatic systems ability
to sustain life. To measure DO content, the Winkler
method was used by titrating the iodine produced by

[5] Pierce, W.C., Haenisch, E.L., Sawyer, D.T.

Quantitative Analysis, 9th ed.; John Wiley & Sons: New
York, 1948.

Chem 31.1 Section, Room No., Time

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[6] Harris, D.C. Quantitative Chemical Analysis, 8th
ed.; W.H. Freeman & Company: New York, 2010.
[7] Skoog, D. A., West, D. M., Holler, F. J., Crouch, S.
R. Fundamentals of Analytical Chemistry, 9th ed.;
Brooks/Cole: Belmont, 2014.

[9] Ohio State University. Common Misconceptions

about
Plants.
http://beyondpenguins.ehe.osu.edu/issue/polarplants/common-misconceptions-about-plants (accessed
October 27, 2015)

[8] Patnaik, P. Deans Analytical Chemistry Handbook,

2nd ed.; McGraw-Hill: New York, 2004.

Chem 31.1 Section, Room No., Time

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