School of Chemical, Biological and Materials Engineering and Sciences

1st Quarter S.Y. 2023-2024

Experiment 2.3
Analysis of Dissolved Oxygen

Submitted by: Date Submitted: 10/21/23

Group No: 1 Signature

Leader: Allen John A. Dalangin

Panopio, Jeremy M. ________________

Magdame, Eamon Miggs C. ________________
Pascua, James Anthony P. ________________
Siazon, Harvey Sean A. ________________

Engr. Elizabeth S. Espiritu

Faculty In-charge
CM011L-SECTION
 School of Chemical, Biological and Materials Engineering and Sciences
1st Quarter S.Y. 2023-2024

1. INTRODUCTION

A. Brief Background

Dissolved oxygen in aquatic environments is vital for aquatic life's existence and
well-being. It is an essential indicator of the water's quality and contributes significantly to
the health and function of aquatic environments. The amount of molecular oxygen (O2)
contained in water is called dissolved oxygen (DO). In this experimental report, the
students were to analyze dissolved oxygen guided with specific objectives: (1) To develop
proper technique for obtaining a natural water sample, (2) To determine the dissolved
oxygen concentration of a natural water sample, and (3) to learn the chemical reactions
involved in fixing and analyzing a water sample for dissolved oxygen using Winkler
Method.

In experimenting, the students first collect a water sample in a bottle and is treated
with manganese sulfate and alkaline iodide. Following the formation of floating particles,
sulfuric acid is added to the bottle, which makes the solution yellowish. 200 mL of the
treated sample is transferred to a separate flask, and adding starch solution continued the
titration results in a blue color. Sodium thiosulfate is then titrated with the solution,
resulting in a color fade/transparency when the titration ends. The dissolved oxygen
content in milligrams per liter (mg/L) is now represented by the amount of sodium
thiosulfate used for titration. This careful procedure ensures precise measurement of
dissolved oxygen levels, critical for measuring water quality and aquatic ecosystem health.
Understanding and monitoring dissolved oxygen levels are essential for maintaining water
environments.

B. Theoretical Discussion

Dissolved oxygen (DO) refers to the concentration of oxygen gas incorporated in

water. Oxygen dissolves in surface water due to the aerating action of winds. Additionally,
aquatic plant photosynthesis introduces oxygen into the water as its byproduct. It is
considered an essential measure of water quality as it directly indicates an aquatic
resource's ability to support marine life (Naresh, 2023).

In the field of aquaculture and fisheries, maintaining an adequate amount of

dissolved oxygen is critical for the survival of aquatic organisms. Most aquatic organisms,
such as fish, invertebrates, and some microorganisms, rely on dissolved oxygen in water
for respiration. Insufficient oxygen can lead to hypoxia or anoxia, where oxygen levels are
too low to support life, causing fish kills and other negative impacts on aquatic
ecosystems. According to the National Aquatic Resources Survey, the appropriate DO
concentration for fish survival is 5 to 6 parts per million (ppm), measured as milligrams per
liter. DO concentrations less than 5mg/L are considered stressful for fish, and
concentrations less than 3mg/L are significantly low to support fish survival. Levels below
1mg/L are hypoxic and usually devoid of life (USEPA, 2023a).

In the field of medicine, dissolved oxygen reflects the percentage of hemoglobin

binding sites in the bloodstream occupied by oxygen. Physicians in medical diagnosis use
oxygen saturation measurements in blood since low saturation or hypoxemia indicates
various medical conditions, including chronic obstructive pulmonary disease (COPD) or
 School of Chemical, Biological and Materials Engineering and Sciences
1st Quarter S.Y. 2023-2024

carbon monoxide poisoning (Nema et al., 2009). For wastewater treatment, dissolved
oxygen is added to enhance the oxidation process by providing oxygen to aerobic
microorganisms so they can successfully turn organic wastes into inorganic byproducts.
For their metabolization and reproduction, microorganisms require 0.1 to 0.3 ppm DO.

Multiple factors affect the concentration of dissolved oxygen in water. Related

factors to DO level fluctuation are temperature, as higher temperature reduces oxygen
solubility; high nutrient concentrations that lead to excessive plant growth, therefore
declining the DO levels due to respiration and decomposition; sediments, which prevent
DO from permeating interstitial areas; and ammonia that consumes oxygen from
nitrification (USEPA, 2023b).

There are multiple ways to measure the concentration of DO in a given water

sample. As oxygen can quickly diffuse into the water sample during handling, samples
must be collected without exposure to air. This ensures that the sample maintains its initial
concentration without alteration during the measurement process. According to
Katznelson (n.d.), the methods of measuring DO levels can be sorted into three major
groups: colorimetric, titrimetric, and polarographic. In colorimetric measurement, the water
sample is sucked into an ampoule under vacuum. Inside, chemical reagents interact with
oxygen to form a colored product. The intensity of the color is proportional to the DO
concentration in the sample. Therefore, it is compared to a series of tubes with color
intensities that reflect known dissolved oxygen concentrations to determine the
appropriate ppm. In polarographic measurement, electrodes measure the flux of oxygen
across a membrane. As oxygen is consumed in the process, traditional electrodes flush
the measured liquid at the membrane surface to constantly replace the oxygen consumed.
Lastly, the titrimetric method measures dissolved oxygen concentration in a sealed water
sample. The dissolved oxygen creates another product using chemical reagents, while
another chemical neutralizes the given concentration. The amount of chemical sufficient to
neutralize the product from the sample is proportional to the DO concentration, which can
be measured in ppm through calculations.

One titrimetric method is the Winkler method. As previously mentioned, a sample

bottle is entirely filled with water and then "fixed" by adding a series of reagents that
create an acid compound that is then titrated with a neutralizing compound that results in
a change of color (Bruckner, n.d.). This method uses 2 mL manganese sulfate, 2 mL
alkali-iodide, 2 mL concentrated sulfuric acid, 2 mL starch solution, and sodium thiosulfate
as the titrant. The procedure is as follows:
 School of Chemical, Biological and Materials Engineering and Sciences
1st Quarter S.Y. 2023-2024

Figure 1. The Process of Winkler Method

This method follows these series of chemical reactions:

2 Mn2+ (aq) + O2 (aq) + 4OH– → 2 MnO(OH)2(s)

MnO(OH)2 (s) + 2 I– (aq) + 4H+ → Mn2+ (aq) + I2 (aq) + 3 H2O

I– (aq) + I2 (aq) → I3– (aq)

2 S2O32- (aq) + I3– (aq) → S4O62- (aq) + 3 I– (aq)

Using the reduction of I- by the titrant, Na2S2O32-, the moles of O2 can be

calculated, measuring its mass and consequently, the present ppm of dissolved oxygen
in the water sample.

2. METHODOLOGY
 School of Chemical, Biological and Materials Engineering and Sciences
1st Quarter S.Y. 2023-2024

The analysis of dissolved oxygen will be the focus of the experiment. This has
several objectives, including learning how to collect a natural water sample properly,
calculating the amount of dissolved oxygen in a natural water sample, and understanding
the chemical process involved in resolving and evaluating a sample of water for dissolved
oxygen by employing the Winkler method. Safety gear (lab coat, gloves, and goggles), a
burette, a burette clamp, an iron stand, a beaker, an Erlenmeyer flask, a bottle, a
graduated cylinder, a glass graduated dropper pipette, tap water, manganous sulfate,
alkaline iodide, sulfuric acid, sodium thiosulfate, and starch solution are the materials used
in this experiment.

Figure 2.1 Tap Water Sample

First, the students execute procedure A, which involves collecting a water sample.
It required a precise method of doing this, letting the faucet run for approximately one
minute at first. After that, letting it run down the sides of the bottle and overflow to prevent
aeration from affecting the experiment.

Figure 2.2 Mixture of Manganous Sulfate and Alkaline Iodide

 School of Chemical, Biological and Materials Engineering and Sciences
1st Quarter S.Y. 2023-2024

Afterwards, the students proceeded with procedure B. By inserting the tip of the
pipette into the solution containers and extracting 5 mL of each manganous sulfate and
alkaline iodide solution into the bottle, excess liquid overflowed which is included in the
procedure. The bottle was then flipped upside down twice for the contents to mix and
allow the precipitation to take place.

Figure 2.3 Addition of Concentrated Sulfuric Acid in the Mixture

After the mixture had precipitated, 5 mL of concentrated sulfuric acid was added in
the same way as the prior solutions. The cap of the bottle was immediately put on, and it
was then gently inverted to mix the contents.

Figure 2.4 Titration of the Sample with Na2S2O3

 School of Chemical, Biological and Materials Engineering and Sciences
1st Quarter S.Y. 2023-2024

For procedure C, the students begin by cleaning the burette with sodium thiosulfate
and letting it run out through the tip, this was done three times The students then filled it
with the same solution, sodium thiosulfate, while making sure there was no air gap at the
tip of the burette. The burette was then fastened to the iron stand, and 200 mL of the
sample and 3 mL of the starch solution were added to the Erlenmeyer flask using the
graduated cylinder. The Erlenmeyer flask was positioned below the tip of the burette, and
sodium thiosulfate was used for titration. The students recorded the data and repeated the
procedure for Trial 2.

3. RESULTS AND DISCUSSION

In this experiment, students successfully analyzed the dissolved oxygen levels in a

natural water sample using the Winkler method. The process involved collecting a 200 mL
tap water sample, adding specific reagents like manganous sulfate and alkaline iodide to
initiate chemical reactions, and performing a titration with sodium thiosulfate to determine
the dissolved oxygen content. The meticulous approach emphasized the importance of
precision and accuracy in environmental water quality analysis. It provided students with
valuable hands-on experience while highlighting the role of chemical knowledge in
environmental studies.

Table 1.1. Water Sample Analysis, Trial 1

Sample Analysis Trial 1

Sample Volume (mL) 200
Initial Buret Reading (mL) 0
Final Buret Reading (mL) 6.6
Volume Na2S2O3 Dispensed (mL) 6.6
Moles of Na2S2O3 Dispensed (mL) 1.65x10-4
Moles of I20 Reduced by S2O32- (mol) 8.25x10-5
Moles of O2 (mol) 4.125x10-5
Mass of O2 (mg) 1.320
Dissolved Oxygen, ppm O2 (mg/L) 6.6

It was determined in trial 1 that the dissolved oxygen concentration in the given
water sample was 6.6 ppm. According to the observation made in the theoretical
discussion, it can be deduced that the water sample can sustain aquatic life, given that the
needed DO concentration is above 5-6 ppm.

Table 1.2. Water Sample Analysis, Trial 2

Sample Analysis Trial 2

Sample Volume (mL) 200
Initial Buret Reading (mL) 0
Final Buret Reading (mL) 6.3
Volume Na2S2O3 Dispensed (mL) 6.3
Moles of Na2S2O3 Dispensed (mL) 1.58x10-4
Moles of I20 Reduced by S2O32- (mol) 7.9x10-5
 School of Chemical, Biological and Materials Engineering and Sciences
1st Quarter S.Y. 2023-2024

Moles of O2 (mol) 3.95x10-5

Mass of O2 (mg) 1.264
Dissolved Oxygen, ppm O2 (mg/L) 6.3

The average DO concentration from the collected water sample is 6.45 ppm. Given
the similar results of both trials, the results can be assumed accurate. This experiment
holds significant environmental implications. Monitoring dissolved oxygen levels is critical
for assessing the health and quality of aquatic ecosystems. Adequate dissolved oxygen is
essential for aquatic life, and deviations from optimal levels can have detrimental effects.
By mastering the Winkler method and its associated chemical reactions, students have
gained the skills needed to contribute to the maintenance and improvement of water
quality, ultimately enhancing the well-being of aquatic environments and the sustainability
of aquatic ecosystems.

4. CONCLUSION

The experiment's core goal is precisely analyzing dissolved oxygen levels in a

natural water sample using the Winkler method. It entails collecting a 200 mL tap water
sample and after mixing it with 5 mL each of manganous sulfate and alkaline iodide
solutions. After precipitation, 5 mL of concentrated sulfuric acid is added. The titration is
performed with 3 mL of starch solution, and sodium thiosulfate is used to assess the
dissolved oxygen. This comprehensive approach not only deepens understanding of the
chemical processes involved but also hones skills needed for exact dissolved oxygen
measurements in environmental water samples, therefore enhancing water quality analysis.

Results showed an average dissolved oxygen concentration of 6.45 ppm in a natural

water sample. This was concluded from the amount of Na2S2O3 dispensed during titration,
which coincidentally averaged an amount of 6.45 mL. However, no direct relation can be
deduced from this observation as the concentration of dissolved oxygen was calculated
through its stoichiometric relation with Na2S2O3. Nonetheless, this means that at a
temperature of 32°, the DO level in a natural water sample is approximately 6.45 ppm. As
stated in the theoretical discussion, aquatic creatures require DO concentrations of 5 -6
ppm, therefore it can be concluded that fishes and alike creature can thrive on the given
water sample. The deliberate procedure used in this experiment helps students develop
critical analytical skills for deciding water quality. The precise use of manganous sulfate,
alkaline iodide, and other reagents in this method emphasizes the value of accuracy and
chemical knowledge in environmental study as it relates to the procedural aspect

REFERENCES:

Brezonik, P. and William A. Arnold, 'Dissolved Oxygen', Water Chemistry: The Chemical
Processes and Composition of Natural and Engineered Aquatic Systems, 2nd edn
(New York, 2022; online edn, Oxford Academic, 17 Nov. 2022),
https://doi.org/10.1093/oso/9780197604700.003.0018

Bruckner, M. (n.d.). The Winkler Method - Measuring Dissolved Oxygen. Montana State
University.
https://serc.carleton.edu/microbelife/research_methods/environ_sampling/
 School of Chemical, Biological and Materials Engineering and Sciences
1st Quarter S.Y. 2023-2024

oxygen.html?
fbclid=IwAR2G5sw2uLqfLQ3f3T5dV135eNBKIKQ1u_caCRow_zbsChh8FOXlJid1g4
o

Katznelson, R. (n.d.). Dissolved Oxygen Measurement Principles and Methods. DQM

Information paper 3.1.1.
https://www.waterboards.ca.gov/water_issues/programs/swamp/docs/cwt/
guidance/311.pdf

Naresh, R. (2023). The Importance of Dissolved Oxygen in Water Quality Parameters and
Importance of Dissolved Oxygen in Waste Water. ResearchGate.
https://www.researchgate.net/post/What_is_the_importance_of_dissolved_oxygen_i
n_water_quality_parameters_and_importance_of_dissolved_oxygen_in_waste_wat
er

USEPA (2023a). Indicators: Dissolved Oxygen. National Aquatic Resource Surveys.

https://www.epa.gov/national-aquatic-resource-surveys/indicators-dissolved-oxygen

USEPA (2023b). Dissolved Oxygen. CADDIS Volume 2. https://www.epa.gov/caddis-

vol2/dissolved-oxygen
 School of Chemical, Biological and Materials Engineering and Sciences
1st Quarter S.Y. 2023-2024