Environmental Chemistry

Laboratory exercises

Dissolved Oxygen

Purpose of the exercise

1. The determination of dissolved oxygen in water samples using Leithe and Winkler methods.
2. Comparison of the results obtained by these two methods.
3. Comparison of the results with the standards for drinking water given in Polish Regulation
currently in force.

1. General consideration
All living organisms are dependent upon oxygen to maintain the metabolic processes
that produce energy for growth and reproduction. Aerobic processes are the subject of greatest
interest because of their need for free oxygen. We are vitally concerned with the oxygen
content of the air that we breathe, since we know from experience that an appreciable
reduction in oxygen content will lead to discomfort in breathing and furthermore possibly to
death. For this reason, the number of occupants within enclosures must be carefully restricted
to the ventilating capacity.
Environmental engineers are interested in atmospheric conditions in relation to humans.
In addition, they are vitally concerned with the "atmospheric conditions" that exist in liquids,
especially water being the liquid of greatest abundance and importance. There are two sources
of oxygen in natural water, namely diffusion from the atmosphere and photosynthesis.
All the gases of the atmosphere are soluble in water to some degree. Both, nitrogen and
oxygen, are classified as poorly soluble. As they do not react chemically with water, their
solubility is directly proportional to their partial pressures. Hence, Henry's law may be used to
calculate the amounts of these gases in water at saturation point at any given temperature. The
solubility of both nitrogen and oxygen varies greatly with the temperature over the range of
interest to environmental engineers. Figure 1 shows solubility curves for the two gases in
distilled or low-solids-content water in equilibrium with air at 760-mm Hg pressure. It can be
noted that under the partial-pressure conditions that exist in the atmosphere, the amount of
nitrogen dissolved in water is higher than the amount of oxygen. At saturation point there is
about 38% of oxygen dissolved in water (on a molar basis), so nearly twice as much as in the
air.
The solubility of atmospheric oxygen in fresh waters ranges from 14.6 mg/L at 0oC to
about 7 mg/L at 35C under 1 atm of pressure. Since oxygen is a poorly soluble gas, its
solubility varies directly with the atmospheric pressure at any given temperature. This issue is
especially important at high altitudes.

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 Figure 1. Solubility of oxygen and
nitrogen in distilled water saturated
with air at 760 mm Hg.
As oxygen demands increase with increasing temperature due to increasing rates of
biological oxidation, high-temperature conditions (where oxygen is least soluble) are of
greatest concern to environmental engineers. Most of the critical conditions related to
dissolved-oxygen deficiency in environmental engineering practice occur during the summer
months when temperatures are high and solubility of oxygen is at a minimum. For this reason
it is customary to think of dissolved-oxygen levels of about 8 mg/L as being the maximum
available under critical conditions.
The low solubility of oxygen is the major factor that limits the purification capacity of
natural waters and necessitates treatment of wastes to remove pollutional matter before
discharge to receiving streams. In aerobic biological treatment processes, the limited solubility
of oxygen is of great importance because it governs the rate at which oxygen will be absorbed
by the medium and therefore also the cost of aeration.
Also the presence of salts alters the solubility of oxygen in water. Solubility in salt-
containing water is lower than in clean water. For this reason the solubility for a given
temperature decreases as one progresses from fresh water to estuary water, and finally to the
ocean. The extent of this effect is indicated in Table 1, which contains a listing of oxygen
solubility as a function of temperature. Chlorides concentration is used as a measure of the
seawater-fresh water mixture in a sample. The chlorides content of seawater is about 19,000
mg/L.
Table 1. Solubility of oxygen in water at different temperatures

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 Another factor influencing oxygen solubility is a degree of water pollution. The rate of
oxygen transfer to polluted waters is lower than to clean water and their ratio is referred to as
the value. In polluted waters also the saturation value of dissolved oxygen (DO) is lower
than in the clean water. The ratio of the saturation value in polluted water to that in clean water
is referred to as the value. They may range as low as 0.4 for and 0.4 for in some
wastewaters. Both and values are important factors in design and selection of aeration
equipment.

2. Environmental significance of dissolved oxygen

In liquid wastes dissolved oxygen (DO) is the factor that determines whether the
biological changes are brought about by aerobic or anaerobic organisms. The former ones use
free oxygen for oxidation of organic and inorganic matter and produce innocuous end
products, whereas the latter bring about such oxidations through the reduction of certain
inorganic salts such as sulfates, and the end products are often very obnoxious. Since both
types of organisms are ubiquitous in nature, it is highly important that conditions favorable to
the aerobic organisms (aerobic conditions) are maintained; otherwise the anaerobic organisms
will take over and create nuisance conditions. Thus, DO measurements are vital for
maintaining aerobic conditions both in natural waters that receive polluted matter and in
aerobic treatment processes intended to purify domestic and industrial wastewater. In the
latter one DO measurements allow to control the rate of aeration but also to prevent excessive
use of air and energy.
DO measurements are also used for a wide variety of other purposes. They are one of
the most important single tests that the environmental engineers use. For example the
maintenance of specified DO levels supports healthy condition for the growth and
reproduction of desired aquatic life. Measurements of DO also serve as the basis of the
biochemical oxygen demand (BOD) test; thus, they are the foundation of the most important
determination used to evaluate the pollution strength of domestic and industrial wastes. As it
is analytically impossible to determine separately every compound contributing to the
pollution of water, then BOD gives the general information about the presence of different
compounds which undergo biochemical oxidation. The rate of biochemical oxidation is
measured by determining residual dissolved oxygen in a system at various intervals of time.
The DO tests serve as the means of control the corrosion. As it is commonly known,
oxygen is a significant factor in the corrosion of iron and steel, particularly in water
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distribution systems and in steam boilers. Removal of oxygen from boiler-feed waters by
physical and chemical means is common practice, especially in the power industry.

3. Methods of determining dissolved oxygen

Originally, the DO measurement was performed by heating samples (to drive out the
dissolved gases) and analyzing the collected gases for oxygen content by methods applied in
gas analysis. Such method required large samples and was very cumbersome and time-
consuming. Nowadays, the Winkler method (called also iodometric method) and its
modifications are the standard volumetric procedures for determining DO. Also membrane
electrodes are used for DO measurements. As the electrodes allow for in situ measurements of
DO, their use has increased significantly since their development.

The Winkler Method

The test depends upon the fact that oxygen (O2) under alkaline conditions oxidizes
manganium ions (Mn2+) to a higher state of valence (Mn4+). Then, under acidic conditions
manganese is capable of oxidizing iodide ions (I-) to iodine (I2). Thus, the amount of I2
released is equivalent to the DO originally present in the sample tested. The iodine is then
measured with standard sodium thiosulfate (Na 2 S 2 O 3 ) solution and interpreted in terms of
DO.
Reactions

If no oxygen is present in the sample, then after addition of sulphate manganese

(MnSO4) and the alkali-iodide reagent (NaOH + KI) a pure, white precipitate of Mn(OH)2
forms (1):

Mn2+ + 2OH- Mn(OH)2 (white precipitate) (1)

Whereas if oxygen is present in the sample, then some of the manganese (Mn2+) is oxidized
(Mn4+), and precipitates as a brown, hydrated manganese oxide (MnO2). The reaction is
usually represented as follows:
Mn2++ 2OH-+ O2 MnO2 + H2 (2)
or

Mn(OH)2 + O2 MnO2 + H2O (3)

The oxidation of Mn2+ to MnO2 is sometimes called a fixation of the oxygen. The
fixation occurs slowly, especially if at low temperatures. Furthermore, it is necessary to move
the flocculated precipitate throughout the solution to enable all the oxygen to react. Therefore,
a vigorous shaking of the samples for at least 20 seconds is needed. In the case of brackish or
seawater, much longer contact time is required.
After shaking the samples for a time sufficient to allow all oxygen to react, the flocs
should have time to settle down so as to leave at least 5 cm of clear liquid below the
stopper. Subsequently a sulfuric acid (H2SO4) is added, providing a low pH conditions. Then
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MnO2 oxidizes I- to produce I2:
MnO2 + 2I- + 4H+ Mn2 + + I2 + 2H2O (4)

Iodine (I2) is rather insoluble in water, but forms a complex with the excess iodide ions to
reversibly form more soluble tri-iodate, thus preventing the escape of I2 from the solution:

I2 + I- I3- (5)

Then, the sample should be stoppered and shaken for at least 10 seconds to allow the
reaction to go up to the completion and to distribute the iodine uniformly throughout the
sample.
The sample is now ready for the titration with sodium thiosulfate (Na 2 S 2 O 3 ):

2 Na 2 S 2 O 3 5H 2 O + I 2 Na 2 S 4 O 6 + 2NaI + 10 H 2 O (6)
2 S 2 O 3 2 - + I2 S 4O62 - + 2 I- (7)

Titration of the sample gives results in mL of titrant, which can be interpreted directly
in terms of mg/L of DO.

Interferring substances
The unmodified Winkler method is prone to interferences of great many substances.
For example, certain oxidizing agents such as nitrites (NO2-) and iron ions (Fe3+) are also
capable of oxidizing I- to I2. Therefore, the results obtained with their presence will be too
high. On the other hand, some reducing agents such as Fe2+, SO32-, S2-, and polythionates,
reduce I2 to I-. Therefore, the results obtained with their presence will be too low. Summing
up, the unmodified Winkler method is applicable only to relatively pure waters.

Membrane electrodes
The membrane electrodes are especially useful to measurements of DO profiles in
reservoirs and streams at various depths. They serve also in biological waste treatment tanks
for monitoring the DO levels. The rates of biological oxygen utilization in such tanks is
determined by placing a sample of the mixed liquor in a BOD bottle and then inserting a DO
membrane electrode to observe the rate at which oxygen is depleted. They can also be used to
obtain rapid dissolved-oxygen measurements as part of the BOD test.
Membrane electrodes are usually calibrated by making measurements in water samples
that have been analyzed for dissolved oxygen by the Winkler procedure. Thus, any errors in
the Winkler analysis will be carried over to the electrode calibration. During dissolved-oxygen
measurements, it is important that sufficient movement of the sample by the electrode be
maintained to prevent low readings which result if oxygen is depleted at the membrane as it is
reduced at the cathode. Membrane electrodes are very sensitive to temperature, and thus either
accurate temperature measurements must be made along with dissolved-oxygen measurements
so that a correction can be applied, or else instruments that are equipped with a thermistor or
other device to compensate automatically for temperature changes must be used.

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4. Procedures

WINKLER METHOD
1. Refill the Winkler bottle with water sample, add 2 mL of the manganese reagent (MnSO4)
and 2 mL of the alkali-iodide (KJ) solution making sure no air is entrained into the
bottle. It is important to just immerse the tips of the dispensers without immersing them
completely. This eliminates possibility of contamination and introduction of an air
bubbles.
2. Mix until suspension will be homogeneous and then leave the sample in dark place until the
sediment of manganese hydroxide drops down.
3. Add carefully 1 ml of H2SO4 (just above the precipitate at the bottom) and close the bottle,
no precipitate can flow out and no air bubble can appear.
4. Mix the sample until entire precipitate will dissolve and then titrate.
5. For that purpose measure 100 ml of the sample to the Erlenmeyer flask and titrate with the
thiosulfate solution to pale yellow color. Then add 1 ml of starch and continue titration to
discoloration of the sample.
6. Calculate dissolved oxygen content [mg/L] in the sample using the following equation:
DO= 0,2 a 1000/ 100
a volume of sodium thiosulfate (Na2S2O3), mL

LEITHE METHOD
The principle of determination: Water sample is treated with a solution of ferrous sulphate in
excess and alkalined by KOH. Oxygen present in water oxidizes ferrous to ferric hydroxide.
The liquid is acidified and the excess of FeSO4 is determined by permanganate.

1. Fill a Winkler bottle with water completely.

7. Slowly add 5 mL of the 0,1N FeSO4 solution (the excess water flows out of the bottle)
making sure no air is entrained into the bottles. It is important to just immerse the tips of
the dispensers without immersing them completely. This eliminates possibility of
contamination and introduction of an air bubbles.
2. Add 5 pellets of KOH.
3. Close the bottle, mix and leave the sample in dark place until the sediment drops down.
4. Add 5 mL of 50 % H2SO4 (just above the precipitate at the bottom).
5. Close the bottle (the excess water will flow out of the bottle), then mix and leave the sample
until the sediment will dissolve and the yellowish color of the solution will disappear.
6. Pour the acidified sample to the 300 mL flask, and immediately titrate with 0.1 N KMnO4
to pale pink color persisting for 10 seconds (b - volume of KMnO4 used for titration of
water samples, mL).
7. Determine the X from the proportion:
5 ml 0,1 N FeSO4 a mL
X ml 0,1 N FeSO4 b mL
where:
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 a a titre of 0.1 N KMnO4 solution
b a volume of 0.1 N KMnO4 solution used for titration, mL
8. Calculate the DO value for the water sample in mgO2/L.
The number of mL of 0.1 N FeSO40.8 mg results in oxygen content in the volume of the
sample.
DO = (5 - X) 0,8 (1000/V)
where:
V - volume of water sample