Clean Water Identification: Determining Dissolved

Oxygen Utilizing Winkler’s Method

V.A. Orcajada1, J.A.B. Bernabe1, and J.R.K. Burce1

1Department of Civil Engineering, Gokongwei College of Engineering
De La Salle University, Taft Avenue, Manila, Philippines
Bachelor of Science in Civil Engineering (BSCE), EV4, GROUP 2
Date Performed: March 02, 2023

ABSTRACT

The quantity of oxygen that has been dissolved in water can be calculated using a method known
as the Winkler method. The volume of oxygen that is dissolved in water, also known as DO, is a crucial
indicator of the overall quality and health of aquatic environments. In this experiment, the standard for
comparison was the drinking water that was provided by the university. The researchers diligently carried
out the Winkler method, which requires the addition of chemicals to an alkaline medium prior to the reaction
with dissolved oxygen, which then acidifies the solution and makes it possible to titrate it. In addition, the
titration was performed twice, and the average concentration of oxygen (O2) was found to be 4.02 mg dm-
3 in both runs. As a whole, the findings indicate that there is a potential for aquatic life to flourish in the tap
water because the oxygen level concentration was within the lower range (4.02<5).

INTRODUCTION In particular, the experiment utilized the

Clean water is crucial to human health,
the economy, and communities. It aids in
providing nourishment, preventing diseases, food
production, and sanitation. Without adequate
access to it, humans would be put at risk of
waterborne diseases such as cholera, typhoid,
dysentery, and diarrhea. Food insecurity might
arise as problems with water availability, which
agriculture is heavily dependent on, could induce
crop failures and food shortages. As such,
collective effort must be taken to guarantee that
water supplies are safe and dependable globally.
Notably, water quality can be determined by the
quantity of dissolved oxygen contained in an
aquatic area.
The volume of dissolved oxygen (DO)
concentration in freshwater systems is directly
correlated to low levels of pollution and high
productivity (Bruckner, n.d.). Moreover, Oxygen
is vital for aquatic animals and plants in order to
respire and metabolize. It also impacts the
biological and chemical processes in water,
including photosynthesis, nutrient cycling, and
decomposition.
Winkler method, which is also recognized as an
iodometric titration introduced by the Hungarian
chemist, Lajos Winkler to portion the quantity of
DO in a water sample (Markgraf, 2019). It does
so by measuring the consumed iodine in the
reaction, which is precisely relative to the
concentration of DO present in a solution.
Moreover, scientists and environmentalists make
use of the process in determining the value of O2,
together with the quality and condition of aquatic
environments (Bruckner, n.d.). The Winkler
method procedure goes by the following chemical
reactions:
(MnSO₄) Manganese Sulfate and
(NaOH+KI) Alkali-iodide-azide are added into a
sampled water to make a white precipitate.
Mn2++2OH- → Mn(OH)₂
However, if O2 (oxygen) is introduced in
the sample, the mixture turns into a brownish
precipitate.
Mn2++2OH- +(X)O₂ → MnO₂+H₂O
 After, the solution is allowed 5 minutes
for oxygen to fully react and the precipitate to
settle. Sulfuric Acid is then added. The acidic
state of the solution will make MnO₂ oxidize to
form iodine, which does not dissolve in water.
Moreover, the iodine reacts with the extra iodide
to make a complex.
MnO₂+2I- +4H+ → Mn2++ I₂+2H₂O
I₂+I- ←→ I₃-
Sodium Thiosulfate is then used to titrate
the solution to reduce the Iodine back to iodide.
The amount of Sodium Thiosulfate used is
directly proportional to the amount of oxygen.
Aside from the Winkler method,
dissolved oxygen can also be calculated with the
amperometric electrode procedure and the use of
optical sensor technology (Zimmermann et al.,
2018). The amperometric electrode method
utilizes the use of a reference electrode,
employed electrode, and an assisting electrode.
The employed electrode, which is usually made
of platinum or graphite, is covered by a
membrane that allows oxygen to be diffused. The
technique works by an electrochemical process
when oxygen molecules in water diffuse over the
membrane to be lowered at a cathode. The
resulting current generated by the reduction is
proportionate to the amount of dissolved oxygen
present in a medium of water. On the other hand,
optical sensor technology determines oxygen
levels by fluorescence quenching. The method
uses a sensor equipped with an oxygen-
permeable tip and an oxygen-sensitive
luminescent dye. When exposed to blue light, the
dye excites and produces a red light. The oxygen
concentration is then measured by how much red
light is emitted.
This experiment aims to understand and
learn Winkler’s method to familiarize its
applications in real-life scenarios. At the end of
the procedure, the researchers will also be able
to calculate and identify the concentration of
dissolved oxygen concentration from the
sampled water.
METHODOLOGY
The researchers conducted the Winkler
Method experiment in the Velasco Hall
Laboratory (VL409) at De La Salle University,
Manila on March 2, 2023. The researchers
sourced the water sample for the experiment from
one of the taps in the laboratory and
characterized it as transparent, odorless, and free
from impurities.
Reagents used include Volume of 2 cm3:
Manganese (II) sulfate solution (MnSO4);
alkaline Potassium iodide solution (KI);
concentrated Sulfuric (VI) acid (H2SO4); and
freshly prepared Starch solution. Volume of 8
cm3: 0.0125 M Sodium thiosulfate solution
(Na2S2O3). Laboratory equipment used: two 150
mL conical flasks; one 250 mL volumetric flask; a
burette with an Iron stand; and a cloth rag for
spillage.
Proceeding with the experiment, tap
water is poured up to the edge in a volumetric
flask and covered. Making use of a pipette, 1.0
cm3 of Manganese (II) sulfate solution is
incorporated into the sample. Similarly, 1.0 cm3 of
alkaline Potassium iodide solution is added as
well. The flask is covered and then inverted to
achieve uniform precipitate distribution in the
sample solution. Once the solution has rested
and precipitate of at least 3 cm below the stopper
has settled, 1.0 cm3 of concentrated Sulfuric (VI)
acid is introduced into the sample solution. The
flask is inverted repeatedly until no precipitate is
visible. Sample solution is put to rest for five
minutes and afterward transferred into two
conical flasks, each containing 100 mL of the
sample solution.
Progressing with the titration process, the
solution is titrated using 0.0125 M Sodium
thiosulphate until golden-yellow color turns to
pale-yellow. 1.0 cm3 of starch solution is then
added to the solution, and thiosulphate solution is
continuously added till deep blue color
disappears, and a clear solution is achieved. The
volume of used thiosulphate solution is noted,
and experiment is repeated for a second trial.
Once all the data from the two trials had been
gathered, the researchers proceeded to compute
the average dissolved oxygen concentration in
the sample in mg dm-3.
The researchers conducted the Winkler
Method experiment procedures in accordance
with the LBYCH1A Laboratory Manual provided
by the Faculty of Chemistry.
RESULTS
The following tables provide the
calculated volume of sodium thiosulfate and the
concentration of DO in the water sample. The
dissolved oxygen concentration in the water
sample was identified through the ratio of
Winkler's method chemical reactions based on
their stoichiometric relationships and then
performed computations. For this experiment,
two trials were conducted. Each made use of 100
mL of the sample solution in which the average of
their DO concentration was determined to be 4.02
mg dm-3. Moreover, a standard deviation of
0.70007 was discovered from the results of the
two trials. It was calculated to determine the
dispersion of data in relation to the mean. Full
solutions and a completed data sheet can be
accessed through the appendices.
Table 1. Calculated Volume of Sodium Thiosulfate
Dispensed for Trial 1 Titration
Initial Na2S2O3 burette reading 28.5 cm3
Final Na2S2O3 burette reading 33.0 cm3
Volume Na2S2O3 dispensed 4.50 cm3
Table 2. Calculated Volume of Sodium Thiosulfate
Dispensed for Trial 2 Titration
Initial Na2S2O3 burette reading 33.0 cm3
Final Na2S2O3 burette reading 36.5 cm3
Volume Na2S2O3 dispensed 3.50 cm3
Table 3. Calculated Concentration of Dissolved
Oxygen in mg dm-3
Trial 1 4.51 mg dm-3
Trial 2 3.52 mg dm-3
Average Concentration of O2 4.02 mg dm-3
Standard Deviation 0.7000714249
Sample Calculation for Trial 1
n(S2O32- ) = [Na2S2O3] x
(Volume Na2S2O3 / 1000)
= 0.0125 M x (4.50 cm3 / 1000)
=5.63 x 10-5 mol
Ratio of O2 to S2O32- = 1:4
n(O2) = n(S2O32-) / 4
= 5.63 x 10-5 mol / 4
= 1.41 x 10-5 mol
m(O2) = n(O2) x Molar Mass O2
= 1.41 x 10-5 mol x 32.00 g mol-1
= 4.51 x 10-4 g x 1000
= 0.451 mg
[O2] = m(O2) / (Volume Sample/1000)
= 0.451 mg / (100 cm3/1000)
= 4.51 mg dm-3
DISCUSSION
A. Rationalization of Method
The Winkler method utilizes iodine
molecules which proportionally bind with oxygen
molecules in a sample of water. Thus, by
measuring the iodine present, dissolved oxygen
can be determined.
The experiment starts with adding water
to overflow in a 250 mL volumetric flask. This is
done to minimize the contamination by the
excess O₂ in the atmosphere. Then, in a similar
manner, Manganese sulfate liquid and alkaline
Potassium iodide liquids are included which
follows the parameter of 2mL/200mL of water.
After, the solution is mixed by inverting the flask
forming a brown precipitate. It is important to “fix”
(convert oxygen to iodine) the sample as soon as
it is gathered as organisms such as
phytoplankton and bacteria can change oxygen
content by respiration (Louisiana Universities
Marine Consortium, n.d.).
Moreover, in the presence of a strong
base, each oxygen molecule forms a manganous
hydroxide complex by binding with a manganese
ion (Louisiana Universities Marine Consortium,
n.d.). The reaction is then allowed for a brief
period for oxygen to fully react and its precipitate
to settle. After, the sulfuric acid concentration is
add up to reduce pH levels and dissolve the
precipitate. Iodine is produced in the process.
(Optionally, starch is included to turn the solution
from light yellow to deep blue which makes for an
easier visualization of color change). The solution
is then split into two 100mL conical flasks to allow
two trials for more accurate results. Lastly, both
solutions are titrated with Sodium Thiosulfate to
produce a controlled chemical reaction and
neutralize the color which ultimately determines
the amount of dissolved oxygen.
B. Purpose of each Reagent
Since Winkler’s method has a process of
indirect titration, each of the chemicals used has
a specific role in determining DO concentration
indirectly. The following are the relevant chemical
reactions that will justify the purpose of each
reagent:
(1) 4Mn2+(aq) + 8OH- (aq) + O2(aq) + 2H2O(l) ⎯→
4Mn(OH)3 (s)
(2) 2Mn(OH)3(s) + 2I-(aq) + 6H+(aq) ⎯→ 2Mn2+(aq) +
I2(aq) + 6H2O(l)
(3) 2S2O32-(aq) + I2 ⎯→ S4O62-(aq) + 2I-(aq)
 Adding manganese (II) sulfate and
alkaline potassium iodide will form a precipitation
reaction of insoluble oxygen manganese
complex. By that state, it will stabilize the O2 in
the water. The accumulation of sulfuric acid to the
solution allows the complex to dissolve, giving a
free iodine component with a golden yellow color.
Note that the quantity of iodine present in the
mixture is proportionate to the volume of DO in
the initial state of the water sample. A
standardized sodium thiosulfate solution is
utilized as a titrant in titrating the solution to
determine the volume of iodine, which also
corresponds to the volume of oxygen. To prevent
any visual errors in titration, a starch solution is
added as a marker to identify the occurrence of
iodine in a deep blue color. With this method, it is Figure 1. Present Air Bubbles during Experiment
simpler to distinguish the endpoint of water from
a deep blue color than a gold-yellow color According to the Water Science School
(Winkler Titration | Flinn Scientific, n.d.) (2019), water areas with a high dissolved oxygen
or at least with a concentration of 3.0 above, may
C. Interpretation of Results have a better quality of aquatic life, enabling
aquatic animals to swim free around without
Based on the calculation, the outcome worrying any poisoning or health related risks. In
reveals that the DO concentration of the sample comparison with the experiment’s results not
tap water from the first trial valued 4.51 mg dm -3 including the instrumental error, the sample water
and for the second trial with 3.52 mg dm-3. With with a mean of 4.02 mg dm-3 is above the
those results, it contained a mean of 4.02 mg dm- minimum concentration of DO. This indicates that
3
with a standard deviation value of 0.7 which is the sample water can contribute to the quality of
less than 1. This indicates that the data points are aquatic life and has sufficient dissolved oxygen
near to each other, giving a reliable result. for some of the aquatic organisms to live in.
However, the results of this experiment may be Contrastingly, an article by Water Quality (n.d.)
undependable due to the instrumental error of argues that 5-6 ppm of DO is typically needed for
having air bubbles during the procedure. The a group of fish to sustain their productivity and
Coyote Creek Riparian Station (n.d.) mentioned development. They claim that most aquatic
that having air bubbles in the experiment will organisms are stressed by water areas having
cause false and high analyses that will alter the dissolved oxygen levels of less than 3 ppm. On
calculations. Taking into account the presence of the other hand, they agree that fish cannot
air bubbles in the experiment, there is a possibility survive at DO levels with less than 2 ppm
that the determined average DO concentration of concentration.
4.02 mg dm-3 is skewed. Focusing on the
outcomes, the quantity of DO in the sampled With that being said, there is still
water from two trials is greater than 3.0 mg dm-3. uncertainty about the findings and may need to
Thus, the water is potentially deemed unpolluted. be reevaluated with little to no errors to ensure
the safeness of the aquatic animals.

As this experiment involves Winkler’s

method, the quantity of dissolved oxygen (DO)
from the sample water is measured by a simple
chemical analysis or a titration-based procedure.
Comparing it to other lab reports, they utilized
different methods with different approaches to
analyzing the DO in a water sample. An article by
Christopher (2019) states that the two other
methods, the Amperometric Membrane Electrode
method, and the Fluorescence method, use
different analyzing procedures. The
Amperometric Membrane Electrode method
employs an electrochemical analysis because it
involves an electrolyte solution by a submergible,
casing-covered electrode that is affixed to a
measuring device that gives direct analyses in
mg/L. On the other hand, The Fluorescence
method applies photochemical analysis because
it includes light wavelength in determining DO.
To further discuss, analyzation of
dissolved oxygen is also relevant in
environmental chemistry research. It can be used
to monitor the impact of human activities on
aquatic environments. To illustrate, factories
discharge several types of untreated chemical
waste that can pollute waters leading to hypoxia.
Under low points of DO, most marine life may
decline in health, undergo acute stress, and even
die (Western Australian Government,
Department of Water, 2015). Furthermore, long
lasting effects range from reduced biodiversity,
ecosystem resilience, to complete alteration of
biogeochemical cycles (Ma et al., 2018b).
Therefore, dissolved oxygen level
analysis in aquatic habitats is an essential
element of environmental chemistry research. It
allows scientists critical information on the health
and quality of aquatic ecosystems and can aid in
the identification of the impact of human activities.
CONCLUSION AND RECOMMENDATION
This experiment aims to identify the
dissolved oxygen concentration in a water
sample through the Winkler Method. The
researchers made use of tap water sample
sourced in one of the laboratories of De La Salle
University Manila. According to the experiment’s
findings, the average concentration of the water
sample was determined to be 4.02 mg dm -3,
which signifies that the water sample can be
deemed unpolluted. Therefore, in case the need
arises, the water sample can be used to host
aquatic life. However, it is not advisable as most
groups of fish experience stress in living
conditions with DO concentrations of less than 5
mg dm-3.
Due to factors such as (1) Presence of air
bubble due to ineffectiveness of volumetric flask
cover used in the experiment; (2) Experiencing
malfunction of burette; (3) Subjectivity of
determining pale yellow color and clearness of
solution in titration; (4) Improper conduct of
experiment that could introduce unwanted
oxygen in the sample, the findings in the
experiment are subject to skewness. To avoid
such from occurring, the following
recommendations are given by the researchers.
To future researchers conducting the
experiment: Make sure to use a rubber stopper if
the volumetric flask provided does not come with
it. This is to ensure that the flask can be tightly
sealed, avoiding presence of air bubbles when
procuring water samples; Inspect laboratory
equipment for flaws before conducting the
experiment. Using faulty equipment can lead to
inaccurate readings; Consult the professor in
charge of the appearance of analyte to avoid
over-titration; Transfer solutions into cylinders
and flasks by making use of a stirring rod to
ensure no air bubbles is incorporated.
To the faculty in charge of laboratory
maintenance: Observe regular laboratory
equipment check-ups. Replace immediately of
defective laboratory equipment. By doing all
provided precautions, optimum accuracy in the
experiment's findings is ensured.
To conclude, the findings indicated in this
lab report show that tap water in the De La Salle
University Manila is considerably clean and to
some extent safe for aquatic life. This observation
bears significance to the fact that access to clean
water is a privilege. Some people in other
countries, even aquatic life, struggle to gain
resources to clean water. That is why the
researchers urge the readers to observe the
collective effort of conserving water. After all,
there is only so much clean water in the world,
and it is only a matter of time before the clean
water we enjoy can no longer be within reach of
our hands.
REFERENCES
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Winkler Method. Environmental
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Coyote Creek Riparian Station. (n.d.).
Measurements of Dissolved Oxygen with
the Modified Winkler Titration.
https://www.waterboards.ca.gov/water_i
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Ma, Z., Wu, H., Zhang, K., Xu, X., Wang, C., Zhu, I hereby certify that I have given substantial
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APPENDICES

APPENDIX A: FILLED OUT DATA SHEET WITH SOLUTIONS

APPENDIX B: METHODOLOGY DIAGRAM