G. Abril 2000, modified by M. Hesselse 2001, A.H.

Nielsen 2007

Oxygen measurement by Winkler titration

In 1888, the Hungarian chemist Lajos Winkler proposed a titremetric method to measure dissolved
oxygen (DO) in waters. Since that time, membrane electrodes have been developed, but Winkler
titrations are still used to calibrate electrodes, and when very accurate measurements are needed.
Winkler titrations can give oxygen values at about 0.05 mg/l (a few M), whereas membrane
electrodes have a precision of approximately 0.1 mg/l and often drift with time. However, in some
particular waters, Winkler titrations are not possible (because of high concentrations of suspended
solids that unable colorimetric titration, high concentrations of organic matter that interfere, etc).

Principle of the Winkler method

The method consists in adding simultaneously a strong base solution containing iodide (Solution 1 =
NaI 3M / NaOH 8M) and a solution of divalent manganese (Solution 2 = MnCl2 3M) which leads to
the precipitation of manganese hydroxide (Mn(OH)2) according to the reaction 1a. The oxygen
dissolved in the water then oxidizes an equivalent quantity of manganese hydroxide into a
manganese hydroxide of higher valence state: MnO(OH)2, according to reaction 1b. By adding a
strong acid, this manganese hydroxide complex is destabilized and, in presence of the iodide (I-)
introduced with the solution 1, this leads to the formation of a quantity of Iodine (I2) equivalent to
the original DO content (Reaction 2). Iodine is then titrated with a standard solution of thiosulfate
(Na2S2O3 0,0040 M) according to reaction 3.

Reaction 1a :

MnCl 2 + 2 NaOH Mn(OH) 2 + 2 Na + + 2 Cl-

Reaction 1b :

O 2 + Mn(OH) 2 MnO(OH) 2

Reaction 2 :

MnO(OH) 2 + H 2SO 4 + 2NaI Mn(OH) 2 + NaSO 4 + H 2O + I 2

Reaction 3 :

I 2 + 2 Na 2S2O3 Na 2S4O6 + 2 NaI

I.e. : each molecule of oxygen (O2) corresponds to 4 molecules of thiosulfate (Na2S2O3).

Standardization of thiosulfate: The molarity of the thiosulfate may vary with time so
standardization is necessary. For that, we use a solution of KIO3. We first add the three reagents to
the water sample, but the other way around (acid > solution 2 > solution 1) so no complex is

formed and no iodine (I2) is produced. We then add a well-known quantity of KIO3 (8 ml 0.0017M)
and I2 is produced by the reaction:

KIO3 + 5 I- + 6 H + 3 I 2 + 3 H 2O + K +

We can then determine the molarity of the thiosulfate by titration (reaction 3).

Preparation of reagents
Solution 1

NaI 3M & NaOH 8M

Dissolve 30g NaI into 50 ml deionized water
Dissolve 16g NaOH into 50 ml deionized water
Let cool down and mix both solutions.

Solution 2

MnCl2:4H2O
Dissolve 60g MnCl2:4H2O in 100 ml deionized water

Concentrated H2SO4

Use gloves and glasses. Be careful.

Thiosulfate (prepare and subsequently determine the exact concentration by titration)

Dissolve 1 g of Na2S2O3:5H2O in deionized water. Add 10 ml of a solution of Na2CO3 (10 g/l) for
conservation. Fill until a total volume of 1000 ml.

Starch indicator

Prepare a thin paste of 1 g soluble starch in a few mL deionized water. Bring 100 mL deionized
water to a boil, remove from heat and stir in the starch paste.

Standard of potassium iodate

Dissolved 0,3567 of dried K(IO3) in 200-300 ml deionized water in a 1L volumetric flask. Fill the
volumetric flask to 1L with deionized water. The molarity is 0.0017 M KIO3.

Standardization of thiosulfate solution

1. Prepare thiosulfate solution as described previously

2. Add approximately 50 mL deionized water to a titration beaker.
3. Add 250 L concentrated sulfuric acid (H2SO4)
4. Add 250 L MnCl2 solution and 250 L (NaI 3 M + NaOH 8 M) solution
5. Add 8.0 mL 0.0017 M KIO3. Mix with magnet.
6. Start titration with the thiosulfate solution (approximately 0.0040 M).
7. Continue the titration until a pale yellow color is obtained. Then add 1-2 drops of starch.
8. Continue titration until the first disappearance of the blue color.
9. Note the amount of thiosulfate added.
10. Calculate the molarity (moles per liter) of the thiosulfate on basis of the titrated volume.

Dissolved oxygen measurement

1. Transfer sufficient sample to the Winkler bottle (approximately 50 or 100 mL) for Winkler
determination of dissolved O2.
2. Fill the Winkler bottle completely.
3. Add 250 L MnCl2 solution and 250 L (NaI 3 M + NaOH 8 M) solution to 50 mL Winkler
bottles (twice the volume to 100 mL Winkler bottles), close and mix. Make sure that no air
bubbles are trapped inside the Winkler bottle.
4. Let the precipitate settle.
5. Add 250 L concentrated sulfuric acid (H2SO4) to 50 mL Winkler bottles (500 L to 100 mL
Winkler bottles). Close and mix until all precipitate has dissolved. At this point, the sample can
be stored for a few hours for later titration.
6. Transfer 50.0 mL to a titration beaker with a glass pipette.
7. Begin titration with the thiosulfate solution.
8. Continue titration until a pale yellow color is obtained. Then add 1-2 drops of starch.
9. Continue titration until the first disappearance of the blue color.
10. Calculate the concentration of dissolved oxygen based on the titrated volume of thiosulfate.

Standardization of Thiosulfate
KIO3 stock
t k solution
l ti 0.0017
0 0017 M
Titration of 8 mL KIO3 solution:
KIO 3 + 5 I- + 6 H+ 3 I2 + 3 H2O + K +

Reaction when adding KIO3 solution:

I.e., 3 moles of iodine (I2) is produced per mole of potassium iodate

(KIO3). Thus, 8 mL KIO3 produces:
8 mL x 3 (mol I2/mol S2O32-) x 0.0017 mol S2O32-/L = 0.0408 mmol I2
Amount of I2 in
titration beaker

Standardization of Thiosulfate
Thi
Thiosulfate
lf t and
d iodine
i di reacts
t 2:1:
21
I2 + 2 Na 2 S 2 O 3 Na 2 S 4 O 6 + 2 NaI

Thus, if x mL of Thiosulfate is added during the titration (until change in

color) the concentration of Thiosulfate is:

Thiosulfate concentration =

mol S 2 O 32 0.0408 mmol I2

mol I2
M
X mL

Thiosulfate concentration =

0.0816 mol S 2 O 32- 0.0816

M
=
X
L
X

Contains
0.0408 mmol I2

Titrate with
x mL Na2S2O3 solution

Thiosulfate standardization example

If 21.1
21 1 mLL Thi
Thiosulfate
lf t was added
dd d during
d i the
th standardization,
t d di ti
the
th
concentration of the Thiosulfate solution is:

Thiosulfate concentration =

0.0816
M = 0.0039 M = 3.9 mM
21.1

Winkler titration with Thiosulfate

A: Thiosulfate
A
Thi lf t concentration
t ti
B: Titrated Thiosulfate volume

=?M
= ? mL

2 /L)
(I M
(I.e.,
Moll S2O32-

The number of moles of Thiosulfate added during the Titration is:

A Mol S2O32-/L x B mL x 1/1000 L/mL

(Unit is Mol S2O32-)

According to the reaction schemes below, Thiosulfate and O2 reacts in

the ratio 4:1.

MnCl2 + 2NaOH Mn(OH)2 + 2Na + + 2 ClO 2 + 2 Mn(OH)2 2 MnO(OH)2

MnO(OH)2 + H2SO 4 + 2NaI Mn(OH)2 + NaSO4 + H2O + I2

I2 + 2 Na 2S 2 O 3 Na 2S 4 O 6 + 2 NaI

Winkler titration with Thiosulfate

The tit
Th
titration
ti iis performed
f
d on 50 mLL off sample;
l th
thus, th
the O2 concentration
t ti can
be calculated from the amount of Thiosulfate added as:

O2 Concentration =

mol S 2 O 321 mol O 2

A
B mL
24 mol S 2 O 3
L
50 mL

If we want the concentration in mg O2/L, we have to multiply with the molar

weight of O2 which is 32 g/mol.

mol S 2 O 321 mol O 2

A
B mL
2L
4 mol S 2 O 3
g
O2 Concentration =
32
50 mL
mol

Winkler titration example

If we added
dd d 12.3
12 3 mLL off 00.0039
0039 M Thiosulfate
Thi lf t solution
l ti d
during
i th
the tit
titration,
ti
then the O2 concentration of our sample was:
O2 Concentration =

mol S 2 O 321 mol O 2

0.0039
12.3 mL
24 mol S 2 O 3
L
g
32
50 mL
mol

O2 Concentration = 0.00768 g O2/L = 7.68 mg O2/L