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Dissolved Oxygen in Water

Chapter · January 2008

DOI: 10.1007/978-0-387-49493-7_2

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Experiment 2
Dissolved Oxygen in Water
Reference Chapters: 6, 7, 8

Objectives oxygen content (or DO level) and the “death” of the

aquatic system. (See Section 8.4.2).
After performing this experiment, the student shall The measurement of the DO is also important
be able to: to determine whether a water system is predomi-
nantly aerobic or anaerobic, predict the survival of
r Determine the level of dissolved oxygen in a sam- aquatic organisms, and predict whether aerobic bi-
ple of water using Winkler’s method. ological processes can take place for transforming
r Analyze the effects of various factors on the level the biodegradable organic contaminants discharged
of dissolved oxygen in a water sample (e.g., salt in water. When there is an organic discharge, the
content, temperature, degree of mixing, and the DO decreases rapidly due to the action of the aer-
presence of reducing compounds). obic microorganisms that consume oxygen during
the metabolic degradation of organic matter (see
Chapter 7). Consequently, the presence of dissolved
oxygen is critical for the self-cleansing of the water
Introduction system, and in combination with the presence of
CO2 , it is also critical for the determination of the
The level of dissolved oxygen in water is one of the corrosive character of water on materials such as
most important parameters in determining its qual- iron and other metals.
ity, because it indirectly indicates whether there is The DO depends on water temperature, dissolved
some kind of pollution. Common processes that pol- salts, atmospheric pressure (and therefore of alti-
lute surface waters include the discharge of organic tude), the presence of reducing compounds, sus-
matter derived from municipal sewage or industrial pended matter, and living species. The aquatic flora
wastes, and runoff from agricultural lots and live- and fauna can contribute to the consumption or to
stock feedlots. In addition, the release of warm or the production of oxygen in water. For the aquatic
hot discharges from industrial cooling towers in- flora, the photosynthesis process is responsible for
duces what is known as thermal pollution. Such dis- the generation of oxygen in the presence of light; this
charges directly affect the level of dissolved oxygen explains the DO fluctuations during the day/night
in water bodies, which is crucial for the survival of cycles and in the different seasons.
aerobic organisms and aquatic fauna such as fish; The presence of microorganisms and biodegrada-
in fact, excessive pollution has caused massive fish tion processes affect the DO level as well. This can
deaths. In the long run, the discharges of organics or also vary vertically, decreasing in the water column
of nutrients favor the accelerated eutrophication or in lakes, large deep rivers, or the ocean due to strat-
productivity process with algal blooms. As a con- ification. By contrast, in rivers and streams it can
sequence, there will be a lowering of the dissolved vary horizontally along the course of the river flow,

16
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Experimental Procedure 17

increasing where there are waterfalls or rapids, and solved oxygen under basic conditions. This step of
decreasing in the slow-moving portions of the river the reaction is called the oxygen-fixation. If there
and in those with organic discharges or microbial is no oxygen present, this step generates a white
activity. Since the dissolution of air in water is an precipitate of Mn(OH)2 ; if there is oxygen present,
interface mass-transfer phenomenon, the degree of a brown precipitate of MnO(OH)2 and MnO2 is
contact and of mixing with water is also important. formed. This precipitate is dissolved with an acid,
The content of oxygen in air is only 21% (v/v) and when iodide ions are present, they are oxidized
and it dissolves in water according to the following to molecular iodine, I2 , which gives a brownish
equation: color to the solution. The amount of liberated io-
dine is directly related to the amount of dissolved
O2(g)  O2(aq) (1)
oxygen. This iodine is titrated with a standardized
This can be represented by Henry’s law (see Section sodium thiosulfate solution, using a starch indicator
6.2.1): for the endpoint. The presence of nitrites can inter-
fere and affect the results noticeably; however, if
K H,O2 = M O2 ,w / p O2 (2) sodium azide is added, it transforms the nitrites into
where N2 and N2 O. The addition of azide is known as the
modified Winkler method.
K H,O2 = Henry’s constant for oxygen In this experiment, students will take several sam-
= 1.29 × 10−3 mol/L-atm (at 298 K) ples of water—some of them saturated with air—
M O2 ,w = [O2(aq) ] = concentration of the dissolved and measure the DO using Winkler’s method, in a
semi micro scale adapted from an original method
oxygen in water, in mol/L
proposed by Ondrus (see Ondrus, 1993).
p O2 (g) = partial pressure of oxygen in air at Because the mixing of reactants must be done
ambient temperature, in atm without introducing additional air to the sample, a
10-mL syringe is used (instead of a standard DBO
Because the partial pressure of oxygen is approx- bottle). In this way, the sampling, the addition of
imated as p O2 (g) = 0.21 Patm (in atm), one can de- reactants, and the reaction all take place in the sy-
termine its concentration in water (in ppm) with this ringe. Another option is to use a small vial of known
relationship by multiplying it times the MW of oxy- volume, equipped with a septum seal, and add the
gen by 1000 (i.e., 32,000): reactants to it with a syringe. At the end, the sample
M O2 ,w (in mg/L) = (K H,O2 ) ( p O2 (g) ) is deposited in a small Erlenmeyer flask and titrated
to the endpoint.
= (K H,O2 ) × 0.21 × 32,000 (3)
The temperature, the mixing and salt content of
Henry’s constant for dilute solutions may approach a water sample are to be modified as well, so as
the real equilibrium constant for the dissolution re- to allow visualization of the factors that affect the
action; however, as the ionic strength of the solution dissolution and the concentration of oxygen in wa-
increases, this is no longer correct, and activities or ter. Finally, to observe the depletion of oxygen by
effective concentrations must substitute concentra- the addition of a reducing compound, one can mea-
tions in the equilibrium ratio. Since oxygen is a non- sure the DO before and after reaction with a re-
electrolyte, it undergoes a salting-out effect where ducing agent. The DO is measured with the same
the water molecules bind to salts, reducing the sol- method described above or with the selective elec-
ubility of the gas. trode method.
One of the two common methods for measur-
ing the DO in water involves an amperometric
method based on a selective electrode for oxygen,
which is the most direct type of measurement. The Experimental Procedure
other, more traditional method involves a volumet-
ric titration—it is known as Winkler’s method and Estimated time to complete the experiment: 15 min
consists in the oxidation of Mn2+ ions by the dis- per sample
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18 2. Dissolved Oxygen in Water

Materials Reagents or samples troduction of extra-dissolved oxygen. In order to

sample correctly, take the sampling bottle or vial
1 10-mL plastic syringe with 0.22 M MnSO4
with a gloved hand and introduce it slowly and hor-
its extreme adjusted with a Alkaline 0.3 M KI and
plastic pipet tip or a piece of sodium azide solution. izontally down the water surface to the height of
Tygon R
tubing and a valve; (Dissolve 5 g of KI, 2.4 g the sampling site. Rinse the bottle with water under
or 1 10-mL bottle adapted of NaOH and 0.3 g of the surface, then fill it completely and cap it inside
with a septum seal NaN3 made up to 100 mL) the water or immediately after it reaches the sur-
1 3- or 1-mL syringe with the 0.018 M H2 SO4 (dissolve
face; make sure that there are no air bubbles trapped
needle cut 1 mL of pure acid to a total
1 100-mL volumetric flask of 100 mL with D.I. water) inside. If samples from deeper levels are needed, a
2 50-mL volumetric flasks 0.00025 M freshly-prepared special sampler must be used.
2 50-mL Erlenmeyer flasks standard Na2 S2 O3 solution
1 Beral pipet sodium metabisulfite, Part A. Measurement of the Dissolved Oxygen
1 25-mm filter holder Na2 S2 O5 (solid)
1 10-mL syringe starch indicator solution Level (D.O.) of an Air-Saturated Sample of
6 25-mL beakers Na2 SO3 (solid) D.I. Water and of a Sample of Tap Water
0.7 μ 25-mm Nitrocellulose NaCl (solid)
filters CoCl2 (solid) 1. Place approximately 50 mL of D.I. water in a
1 microburet (e.g., a 2-mL tap water 100-mL beaker and saturate it with air by aeration
1/100 graduated pipet with a D.I. water with an air pump through a diffuser, for at least
3- or 5-mL syringe adapted river or lake water 20 minutes. Write down the ambient temperature.
with latex or Tygon R

tubing; or an insulin syringe

2. In the meantime, apply Winkler’s method to a
adapted with a thin Tygon R tap water sample. This technique will be used
tubing, see Figure I) throughout the experiment, with all samples.
1 wash bottle with D.I. water 3. After aeration, determine the D.O. level in the
3 air pumps (e.g., aquarium air saturated D.I. sample using the same technique.
pump) equipped with Tygon
tubing and a diffuser
5 100-mL beakers
Steps in the Microscale Winkler’s Technique
1 support stand and clamps for
the microburet 1. Rinse the 10-mL syringe prepared as shown in
1 ice bath
Figure 1 by pulling the plunger and slowly draw-
3 thermometers or
thermocouples ing approximately 2 mL of the water sample.
2 hot plates with sand bath Then, turn the syringe upwards and tap the body
1 stir plate of the syringe softly so as to liberate the small
1 magnetic stirrer bubbles on the surface of the plastic material.
1 small spatula
1 25-mL graduated cylinder
1 bottle for residues
1 10-mL graduated pipet
1 weighing balance

Safety Measures Syringe

Sulfuric acid is corrosive and contact with the skin

or eyes must be avoided. All the residues generated
in this experiment must be disposed of in a special Clasp valve Septum
container identified for that purpose. 10-mL
Before proceeding with the experiment, obtain bottle or vial
the value of the current barometric pressure.
Note: An incorrect sampling process in natural wa-
ters can alter the results considerably due to the in- FIGURE 1. Adapted syringe or vial for reaction.
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Experimental Procedure 19

Let them buoy to the tip and press the plunger c. Carefully draw 2–3 mL of 0.018 M sulfuric acid
to release them from the syringe. Now turn the into the reaction chamber in the same manner de-
syringe downwards and slowly push the plunger scribed above. Shake the entire reaction mixture
to expel the entire sample. This procedure will carefully but thoroughly until the precipitate dis-
leave a small amount of water filling the tip of solves in the acid medium. If a small residue of
the syringe. If the plunger is then softly pushed precipitate remains, add a very small amount of
further, a small drop of water will form at the tip extra acid so as to dissolve it.
of the syringe.
2. Now take approximately 6 mL of the target water (Note: After adding sulfuric acid, the presence of air
by introducing the tip of the syringe deep inside in contact with the sample will not affect the results).
the water sample in the beaker, and slowly pulling The color of the final solution (from pale yellow
out the plunger. No visible air bubbles, except to brown) will depend on the amount of dissolved
those of the sample, must be contained in the oxygen initially present in the sample. The darker
syringe. the color, the higher the amount of initial oxygen
3. Next, incline the syringe tip upwards and slowly dissolved. A brown color reflects the presence of I02
push the plunger in, so as to expel the necessary (in the form of I−3 ).

volume and leave exactly 5 mL of sample inside. Once the reaction is complete (i.e., the precipitate
If a vial is used instead of the syringe, fill it to is dissolved), dispense the content of the syringe into
capacity (record the exact volume added), and a clean, 50-mL Erlenmeyer flask. Use 3–5 mL of
seal it with a septum. D.I. water to rinse the body of the syringe and add it
to the sample in the flask. Fill a microburet with the
Care must be taken in order to prevent any air 0.00025 M Na2 S2 O3 solution and titrate the iodine
bubbles from getting into the syringe or vial as the in the sample (liberated as I−3 ) to a yellowish color.
following reactants are introduced: Then add two to three drops of starch indicator. A
green to blue color must develop. Softly, swirl the
a. Draw 0.5 mL of the manganese sulfate solution
Erlenmeyer flask and continue titrating slowly to a
into the syringe or vial (hereafter called the reac-
colorless endpoint. Record the total volume of titrant
tion chamber). Take care not to introduce more
used.
than what is specified, because once the solution
is inside, it cannot be taken out. If you use a sy-
ringe, close its tip using a septum, or by either Part B. Dissolved Oxygen Measurement of an
bending or clasping tightly a piece of tubing at- Aerated D.I. Sample Pre-treated with a
tached to the tip. If you use a vial, introduce the Reducing Agent
reactant with the syringe. In either case, after that,
shake the system up and down several times to Take a sample of 20 mL of the aerated D.I. water,
mix the contents well. place in a beaker, and with a thin spatula add a very
b. Draw 0.5 mL of the alkaline iodide-azide solu- small amount of a reducing agent, that is, sodium
tion into the syringe or vial in the same manner as sulfite or sodium metabisulfite, and a small crystal
in the previous step. The plunger tip must now be of cobalt (II) chloride as catalyst. Let the mixture
at the 6 mL mark. Once more close the tip of the react, then take a filtered sample of the treated water
syringe and mix thoroughly as mentioned above and apply the same technique as before to measure
(i.e., allow it to react mixing the contents by shak- the dissolved oxygen. Observe the result after the
ing the reaction chamber up and down at least 10 reducing treatment, and compare it with the D.O. of
times or for 1 or 2 minutes). Observe the kind the original sample (i.e., aerated D.I. water).
of precipitate formed. If it is cloudy-brownish,
this signals the presence of manganese hydroxide
Part C. Effect of Temperature in the
(III), which also indicates that there is dissolved
Dissolved Oxygen Level in Water
oxygen present. If it is a cloudy-white precipi-
tate, then this means that no dissolved oxygen is Prepare three different samples of D.I. water of
present. 50 mL each in a beaker, and place them in different
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20 2. Dissolved Oxygen in Water

temperature baths: for example, (a) one in an Part E. Dissolved Oxygen Measurement of
ice bath (∼4◦ C), (b) a second one at 40◦ C and, Two Samples of Surface Water (River or
(c) another at 80◦ C. These last two may be pre- Lake): One Fixing the Oxygen on Site and the
pared in a sand bath, each at different temperature. Other, After Transporting it to the Laboratory
With the air pump let air bubble into the separate
samples in their corresponding baths, for at least 1. Carry out the measurement of dissolved oxygen
10 minutes each. After this, determine the tem- with the modified Winkler technique of a river or
perature, take a sample of each, and measure the lake surface water sample. The most reliable re-
dissolved oxygen level using the modified Winkler sults are obtained if the sample is processed and
technique. the dissolved oxygen is fixed through the first
two reactions of the Winkler method at the site
of sampling, after which the processed sample
Part D. Effect of Ionic Strength in the
is carefully transported to the laboratory for the
Dissolved Oxygen Level in Water
titration process (three samples for more relia-
Because the ionic strength or salt content will affect bility). The sampling may be done with a vial
the dissolved oxygen level, prepare 100 mL of a or directly with the syringe. That is, carry out
blank solution containing 50 g/L of NaCl. Prepare the first three steps of the Winkler technique, and
three beakers: (1) in one beaker, put 50 mL of the leave the titration step for the laboratory.
blank solution; (2) in a 50-mL volumetric flask, di- 2. Take another surface water sample at the same
lute 30 mL of the blank solution to 50 mL; (3) pre- site and this time do not fix the oxygen (i.e., do
pare another 50 mL in a volumetric flask with 15 mL not add any reactants); only take care to cap the
of the blank solution diluted with D.I. water. Place sample well until it is measured in the laboratory.
each solution in a beaker and saturate with air for Then, take the capped sample to the laboratory
at least 15 minutes, and then take a sample and and carry out the Winkler technique as specified.
measure the D.O. with the same modified Winkler After doing this, you will see the difference in
method. the measurements on site and off site.
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Experimental Procedure 21

Name Section Date

Instructor Partner

PRELABORATORY REPORT SHEET—EXPERIMENT 2

Objectives

Procedure flow sheet

Waste containment procedure

PRELABORATORY QUESTIONS 6. Establish all the chemical reactions that take place
in the modified Winkler Method.
1. Explain why the dissolved oxygen content of an
environmental sample of water is important. 7. Explain what is understood as the ionic force of
a solution, what determines it, and how it can be
2. Look up the reported limit values for dissolved
calculated.
oxygen in water so that a system may support
aquatic life. What would be the limit value for a 8. Explain the concept of salting out and how it af-
system to still be considered aerobic? Anaerobic? fects the solubility of non-electrolytes.
3. Explain what contributes or favors an increased 9. What other concept and units besides mg/L is
value of dissolved oxygen in water. used to express the presence of dissolved oxygen in
4. Which are the main types of pollutants that affect water? How is this calculated?
the level of dissolved oxygen in water?
10. Using the theoretical approach, calculate the
5. In what cases would it be undesirable for dis- saturation concentration of dissolved oxygen under
solved oxygen in water to be present? standard conditions.
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22 2. Dissolved Oxygen in Water

11. Using the theoretical approach, explain how r An additional experiment can be carried out in
you would calculate the concentration of dis- which an air-saturated water sample properly
solved oxygen at other latitudes and at other seeded with activated sludge is dosed, and the
temperatures. D.O. is measured with time.

r Another experiment would be to add a small

Additional Related Projects
amount of a biodegradable organic substrate to
r Consider allowing the reduced sample to re-aerate an air-saturated water sample seeded with acti-
and measure the D.O. level with time in order vated sludge and to measure the level of dis-
to determine when the reducing agent is con- solved oxygen vs. time (preferably with the
sumed and how fast the saturation level is attained amperometric method). Winkler’s method has the
again. disadvantage of being a destructive test, and there-
fore the amount of mixture or sample would
r Consider using in the experiment samples have to be large enough to allow the mea-
from other natural systems, for example, ocean surements, yet not interfere too much with the
water. process.
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Experimental Procedure 23

Name Section Date

Instructor Partner

LABORATORY REPORT SHEET—EXPERIMENT 2

Part A. Measurement of the dissolved oxygen level (D.O.) of an air-saturated sample of

D.I. water and a direct sample of tap water

Experimental data

Sample A#1: Air-saturated D.I. water

Temperature:

Barometric pressure:

Sample volume used:

Color of the observed precipitate after the first two reactant additions:

Sample A#2: Tap water

Temperature:

Barometric pressure:

Sample volume used:

Color of the observed precipitate after the first two reactant additions:

Titrant: Titrant concentration:

Sample mL of titrant D.O. concentration, mg/L

Air-saturated D.I. water
Tap water

Formula used:
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24 2. Dissolved Oxygen in Water

Part B. Dissolved oxygen measurement of an aerated D.I. sample treated with a

reducing agent
Experimental data

Sample B#1: Aerated D.I. sample treated with a reducing agent

Temperature:

Barometric pressure:

Sample volume used:

Reducing agent:

Color of the observed precipitate after the first two reactant additions:

Titrant: Titrant concentration:

Sample mL of titrant D.O. concentration, mg/L

B#1

How does the value compare with the two previous samples? Was the reducing agent efficient?

Part C. Effect of temperature in the dissolved oxygen level in water

Barometric pressure:

Sample volume used:

Titrant: Titrant concentration:

Sample D.O. concentration,

Sample temperature, ◦ C mL of titrant mg/L
A#1
C.a
C.b
C.c

How do the values vary with temperature?

Part D. Effect of ionic strength (or salt content) in the dissolved oxygen level in water
Experimental data

Barometric pressure:

Sample volume used:

Titrant: Titrant concentration:

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Experimental Procedure 25

Theoretical
Conc. NaCl, ionic strength, D.O. concentration,
Sample mg/L I (mol/L) mL of titrant mg/L
A#1 0 0
D.a
D.b
D.c

Formula used and calculations for the ionic strength:

Draw a graph of D.O. concentration as a function of ionic strength.

Part E. Dissolved oxygen measurement of two samples of river or lake surface water:
1) Fixing the oxygen on site, and 2) after transporting it to the laboratory.
Experimental data

Sample E#1: Surface water measured on site

Source:

Description of site and place (level) of sampling:

Sampling conditions or precautions:

Temperature:

Barometric pressure:

Sample volume used:

Color of the observed precipitate after the first two reactant additions:
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26 2. Dissolved Oxygen in Water

Sample E#2: Surface water measured at the laboratory

Source:

Description of site and place (level) of sampling:

Sampling conditions or precautions:

Temperature:

Barometric pressure:

Sample volume used:

Color of the observed precipitate after the first two reactant additions:

Titrant: Titrant concentration:

% saturation with
D.O. concentration, respect to saturated
Sample mL of titrant mg/L D.I. water
E#1
E#2

POSTLABORATORY PROBLEMS 2. Explain how you calculated the saturation per-

AND QUESTIONS centage of the samples.

1. Does the value of the measured parameter differ 3. Is each D.O. level acceptable for aquatic life?
from one sample to another? What may be the causes Why?
of such differences? 4. Why is it important to measure this parameter on
site?
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Experimental Procedure 27

Student Comments and Suggestions Literature References

Ondrus, M. G. Laboratory Experiments in Environmen-
tal Chemistry; Wuerz Publishing: Winnipeg, Canada,
1993.
Sawyer, C. N.; McCarty, P. L.; Parkin, G. F. Chemistry
for Environmental Engineering, 5th ed.; McGraw Hill:
New York, 2003.
Stumm, W.; Morgan, J. J., Aquatic Chemistry: Chemical
Equilibria and Rates in Natural Waters; Wiley Inter-
science: New York, 1996.

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