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Determination of Oxygen dissolved in water by Winkler’s Method

History

The test was first developed by Lajos Winkler while working on his doctoral dissertation
in 1888. The amount of dissolved oxygen is a measure of the biological activity of the
water masses. Phytoplankton and macroalgae present in the water mass produce oxygen
by way of photosynthesis. Bacteria and eukaytotic organisms (zooplankton, algae, fish)
consume this oxygen through respiration. The result of these two mechanisms determines
the concentration of dissolved oxygen, which in turn indicates the production of biomass.
The difference between the physical concentration of oxygen in the water (or the
theoretical concentration if there were no living organisms) and the actual concentration
of oxygen is called the biological demand in oxygen.

Principle: The Winkler test is used to determine the level of dissolved oxygen in water
samples and to estimate the biological activity in the water sample. An excess of
Manganese(II) salt, iodide (I-) and hydroxide (OH-) ions are added to a water sample
causing a white precipitate of Mn(OH)2 to form. This precipitate is then oxidized by the
dissolved oxygen in the water sample into a brown Manganese precipitate. In the next
step, a strong acid (either hydrochloric acid or sulphuric acid) is added to acidify the
solution. The brown precipitates then convert the iodide ion (I-) to Iodine. The amount of
dissolved oxygen is directly proportional to the titration of Iodine with a thiosulphate
solution.

Method First Manganese(II) sulfate is added to an environmental water sample. Next,


Potassium iodide is added to create a pinkish-brown precipitate. In the alkaline solution,
dissolved oxygen will oxidize manganese(II) ions to the tetravalent state.

2 Mn(OH)2(s) + O2(aq) → 2 MnO(OH)2(s)

MnO(OH)2 appears as a brown precipitate. There is some confusion about whether the
oxidised manganese is tetravalent or trivalent. Some sources claim that Mn(OH)3 is the
brown precipitate, but hydrated MnO2 may also give the brown colour.

4 Mn(OH)2(s) + O2(aq) + 2 H2O → 4 Mn(OH)3(s)

The second part of the Winkler test reduces acidifies the solution. The precipitate will
dissolve back into solution. The acid facilitates the coversion by the brown, Manganese-
containing precipitate of the Iodide ion into elemental Iodine.

The Mn(SO4)2 formed by the acid converts the iodide ions into iodine, itself being
reduced back to manganese(II) ions in an acidic medium.

Mn(SO4)2 + 2 I-(aq) → Mn2+(aq) + I2(aq) + 2 SO42-(aq)

Thiosulfate solution is used, with a starch indicator, to titrate the iodine.


2 S2O32-(aq) + I2 → S4O62-(aq) + 2 I-(aq)

Analysis

From the above stoichiometric equations, we can find that:

1 mole of O2 → 4 moles of Mn(OH)3 → 2 moles of I2

Therefore, after determining the number of moles of iodine produced, we can work out
the number of moles of oxygen molecules present in the original water sample. The
oxygen content is usually presented as mg dm-3.

Application
Dissolved oxygen analysis can be used to determine:
• the health or cleanliness of a lake or stream,
• the amount and type of biomass a freshwater system can support,
• the amount of decomposition occurring in the lake or stream.

Limitations

The success of this method is critically dependent upon the manner in which the sample
is manipulated. At all stages, steps must be taken to ensure that oxygen is neither
introduced to nor lost from the sample. Furthermore, the water sample must be free of
any solutes that will oxidize or reduce iodine.

Procedure

1. Carefully fill a 300-mL glass Biological Oxygen Demand (BOD)


stoppered bottle brim-full with sample water.
2. Immediately add 2mL of manganese sulfate to the collection bottle by
inserting the calibrated pipette just below the surface of the liquid. (If the
reagent is added above the sample surface, you will introduce oxygen into
the sample.) Squeeze the pipette slowly so no bubbles are introduced via the
pipette.
3. Add 2 mL of alkali-iodide-azide reagent in the same manner.
4. Stopper the bottle with care to be sure no air is introduced. Mix the
sample by inverting several times. Check for air bubbles; discard the sample
and start over if any are seen. If oxygen is present, a brownish-orange cloud
of precipitate or floc will appear. When this floc has settle to the bottom, mix
the sample by turning it upside down several times and let it settle again.
5. Add 2 mL of concentrated sulfuric acid via a pipette held just above the
surface of the sample. Carefully stopper and invert several times to dissolve
the floc. At this point, the sample is "fixed" and can be stored for up to 8
hours if kept in a cool, dark place. As an added precaution, squirt distilled
water along the stopper, and cap the bottle with aluminum foil and a rubber
band during the storage period.
6. In a glass flask, titrate 20 mL of the sample with sodium thiosulfate to a
pale straw color. Titrate by slowly dropping titrant solution from a calibrated
pipette into the flask and continually stirring or swirling the sample water.
7. Add 2 mL of starch solution so a blue color forms.
8. Continue slowly titrating until the sample turns clear. As this
experiment reaches the endpoint, it will take only one drop of the titrant to
eliminate the blue color. Be especially careful that each drop is fully mixed
into the sample before adding the next. It is sometimes helpful to hold the
flask up to a white sheet of paper to check for absence of the blue color.
9. The concentration of dissolved oxygen in the sample is equivalent to the
number of milliliters of titrant used. Each mL of sodium thiosulfate added in
steps 6 and 8 equals 1 mg/L dissolved oxygen.

Result Analysis

The total number of milliliters of titrant used in steps 6-8 equals the total dissolved
oxygen in the sample in mg/L. Oxygen saturation is temperature dependent - gas is more
soluble in cold waters, hence cold waters generally have higher dissolved oxygen
concentrations. Dissolved oxygen also depends on salinity and elevation, or partial
pressure.