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.
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.
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.
Analysis
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
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.