2.
Measurement of temperature, Turbidity/ penetration of light, determination of pH, dissolved Oxygen
Content (Winkler’s method) RAMJAS COLLEGE, UNIVERSITY OF DELHI
Aim: To estimate the quantity of dissolved oxygen present in water by Winkler’s method.
Requirements: Corical flask, Measuring cylinder, funnel, Dropper, Pipettes, Sampling Water
Bottle, Burette with stand. Chemicals include the following: -
1. MnSO4 (36.4 gms in 100 ml of distilled water)
2. NaOH (50 gms) or KOH (70 gms) and NaI (13.5 gms) or KI (15 gms) in 100 ml distilled
water, used as alkali solution.
3. 2% starch solution (2 gms in 100 ml)
4. Conc. H2SO4
5. Sodium Thiosulphate (N/80)
Stock solution: - 24.8 gms in 100 ml of distilled water and preserved by 5 ml chloroform at 40C.
Working solution: - 12.5 gms in 1 litre of distilled water i.e N/80
Theory: - Oxygen is of paramount importance to all living organisms in any aquatic ecosystem.
The occurrence of dissolved oxygen in water is mainly attributed to two distinct phenomena: -
i. Direct diffusion from the air and
ii. Photosynthetic evolution by aquatic autotrophs.
The first one is purely a physical process and depends upon the solubility of oxygen under the
influence of temperature, salinity, water movement, etc., whereas the latter is a biological
process and depends on the availability of light and rate of metabolic processes resulting in
diurnal fluctuations.
Principle: The modified Winkler’s method works on the procedure of titrimetry. Although, it
is a little time consuming, it enables the storage of sample and has precision and accuracy.
The oxygen present in water combines with Manganous Sulphate to form higher hydroxides,
which on acidification, liberates iodine equivalent to that of O2 fixed. This iodine is titrated by
standard sodium thiosulphate using starch as an indicator. The chemistry of the test which is
based on two oxidation seduction reactions is as follows: -
2. Measurement of temperature, Turbidity/ penetration of light, determination of pH, dissolved Oxygen
Content (Winkler’s method) RAMJAS COLLEGE, UNIVERSITY OF DELHI
- Sodium hydroxide reacts with manganous sulphate to give a white precipitate of
manganous hydroxide.
MnSO4 + 2NaOH → Mn(OH)2 + Na2SO4.
- In the presence of oxygen, in a highly alkaline solution, the white manganous hydroxide is
oxidized to brown manganese oxyhydrate.
2Mn(OH)2 + O2 → 2MnO(OH)2.
The white precipitate is then oxidised by the oxygen that is present in the water sample into a
brown manganese-containing precipitate with manganese in a more highly oxidised state (trivalent
or tetravalent)
- Upon the addition of the acid, the precipitate is dissolved forming manganic sulphate.
Mn0(OH)2 + 2H2SO4 → Mn (SO4)2 + 3H2O
- There is immediate reaction between this compound and potassium iodide previously
added, liberating iodine.
Mn (SO4)2 +2KI → MnSO4 + K2SO4 + I2
- This iodine liberated is equivalent to oxygen present and is measured by titrating with
sodium thiosulphate using starch as an indicator.
2Na2S2O3 + I2 → Na2S4O6 +2NaI
Procedure:-
1. Take 300 ml of water in the bottle.
2. Add 2 ml MnSO4 and 2 ml of alkaline KI to water in the bottle.
This white precipitate is then oxidized by the oxygen that is present in the water sample into a
brown manganese – containing precipitate with manganese in a more highly oxidized state
(trivalent or tetravalent)
2. Measurement of temperature, Turbidity/ penetration of light, determination of pH, dissolved Oxygen
Content (Winkler’s method) RAMJAS COLLEGE, UNIVERSITY OF DELHI
The basic steps of measuring dissolved oxygen by the Azide-Winkler titration method.
Stopper the bottle and mix the reagents by inverting the bottle several times.
3. Then add 2 ml of conc. H2SO4 and mix it gently.
4. From this measure 100 ml of the solution in a conical flask and add 1 ml of starch solution
(indicator solution).
5. Titrate it with N/80 sodium thiosulphate till the first disappearance of blue color to
colourless solution.
Result:-
The dissolved oxygen present in tap water of 4.5 mg/L, while that present in pond water is 13.4
mg/L.
Discussion:-
2. Measurement of temperature, Turbidity/ penetration of light, determination of pH, dissolved Oxygen
Content (Winkler’s method) RAMJAS COLLEGE, UNIVERSITY OF DELHI
Dissolved oxygen analysis can be used to determine: -
a. the health or cleanliness of a water body.
b. the amount and types of biomasses which a freshwater system can support.
c. the amount of decomposition occurring in the water body.
The amount of dissolved oxygen is a measure of biological activity of the water masses.
Phytoplankton and macroalgae present in the water body produce oxygen by way of
photosynthesis. Bacteria and eukaryotic organisms (zooplankton, algae and fish) consume
this oxygen through respiration. The result of these two mechanisms determines the
concentration of dissolved oxygen, which in turn, indicates the biomass production. The
amount of oxygen also determines the level of pollution in aquatic ecosystem because it
decreases with the increase of pollution. If water is polluted with organic matter, O2
consumption greatly exceeds O2 production, resulting in depletion of oxygen and
eventually resulting in anaerobic respiration.
The amount of oxygen present in any water body also depends upon a number of
other factors like temperature, salinity, etc. As the temperature increases, the ability of
water to hold dissolved oxygen decreases. The amount of dissolved oxygen reaches its
maximum level in the late afternoon and fall again at night because of absence of
photosynthesis and decrease in the temperature. This rise and fall in the oxygen level within
a day is termed as oxygen pulse.
Precautions:-
1. All the apparatus used should be clean.
2. There should be no air bubble in the burette as well as in the sampling bottle.
3. The solution of the burette should be added dropwise near the end point, with constant
swirling of the conical flask.
4. End point reading should be taken carefully.
5. Three concordant readings should be taken.