WORK INSTRUCTION

Dissolved Oxygen in water

Winkler method (azide modification)

Purpose and Scope

This work instruction is to describe the steps used for the determination of dissolved oxygen content in
effluent water.

Principle
Oxygen present in sample rapidly oxidises the dispersed divalent manganous hydroxide to its higher
valency, which is precipitated as a brown hydrated oxide after the addition of NaOH/KOH and Kl.
Upon acidification, manganese reverts to divalent state and liberates iodine from Kl equivalent to the original
DO content. The liberated iodine is titrated against original DO content. The liberated iodine is titrated against
standard thio sulution using starch as an indicator.
1 MnSO4 + 2KOH ® Mn(OH)2 + K2SO4(white ppt)
2 2 Mn(OH)2 + O2 ® 2 MnO(OH)2 (Brown ppt)
3 MnO(OH)2 + 2H2SO4 + 2 Kl ® MnSO4
4 Mn(SO4)2 + 2 Kl ® MnSO4 + K + l2
5 2Na2S2O3.5H2O + l2 ® Na2S4O6 + 2NaCl + 10H2O
6 2NaN3 + H2SO4 ® 2HN3 + Na2SO4
7 HNO2 + HN3 ® N2 + N2O + H2O

Special Equipment
Sl. No. Equipment
1 Titration assembly & BOD bottles

Reagent/Solution
Sl. No. Reagents
Manganous sulfate solution 480g MnSO4.4H2O or 400g MnSO4.2H2O or 364g MnSO4.H2O in
1
1000ml DM
Alkali-iodide-azide reagent 500g NaOH or 700g KOH and 135g NaI or 150g KI in 950ml DM.
2 Potassium and sodium salts may used interchangeably. Add 10g
NaN3 dissolved in 40ml DM.
3 Sulfuric acid conc. 1ml = 3ml alkali -iodide-azide reagent
4 Starch 2g soluble starch and 0.2g salicylic acid in 100ml hot DM
standard sodium thiosulfate 6.205g Na2S2O3.5H2O + 0.4g NaOH in 1000ml DM.Standardise with
5
(0.025N) bi-iodate solution. 1 ml = 0.2mg of O2

Activities: Sample Analysis

sample in BOD bottle +1ml MnSO4 solution+1ml alkali-iodide-azide reagent, mix by inverting bottles few times,
allow to settled (DO fixation), add 1.0ml conc. H2SO4, restopper and mix by inverting until dissolution of
precipitates, titrate 201ml sample against standard sodium thiosulfate using 2ml starch indicator.

Calculation
DO (mg/l) = 0.2 X 1000 X ml of thiosulphate / 200

*Note: Always wear cotton apron, safety Shoes, gloves and goggles in working area.*
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