EXPERIMENT 5

DETERMINATION OF DISSOLVED OXYGEN IN WATER USING THE

WINKLER METHOD

1.0 OBJECTIVE

i. To determine the dissolved oxygen concentration in water sample

ii. To learn the chemical reactions involved I the Winkler dissolved oxygen method

2.0 INTRODUCTION

Oxygen 𝑂2 is colourless, odourless and tasteless diatomic gas. Oxygen is poorly soluble
in water. Dissolved oxygen (DO) is important parameter in assessing water quality
because the quantity of oxygen in water provide a good condition or circumstances for
aquatic living to life.

There are two methods which are common used to determine the concentration of
dissolved oxygen on water samples: The Winkler (Iodometric) method and the membrane
electrode technique. Membrane electrode is based on rate of diffusion of oxygen
molecules through the membrane. Winkler method is a titration-based method.
Iodometric method is more accurate and it determines the DO concentration through a
series oxidation-reduction reaction.

First, 𝑀𝑛𝑆𝑂4 is added to a 250mL water sample where inside the Biological Oxygen
Demand (BOD) stoppered bottle. Then, alkali-iodide reagent is added. Under this caustic
condition, 𝑀𝑛2+ in 𝑀𝑛𝑆𝑂4 will oxidized to 𝑀4+ . The reaction is relatively slow and
require to shake several times to complete the reaction brown precipitate is form
(hydrated oxide)

2𝑀𝑛2+ + 4𝑂𝐻 − + 𝑂2 → 2𝑀𝑛𝑂2 (𝑠) + 𝐻2 𝑂 (1)

After 𝑀𝑛𝑂2 𝑝𝑒𝑟𝑐𝑖𝑝𝑖𝑎𝑡𝑒 settles to the bottom of the bottle, sulphuric acid 𝐻2 𝑆𝑂4 is added.
Under these low-pH conditions, 𝑀𝑛𝑂2 act as reducing agent oxidizes the iodide 𝐼 − to free
iodine 𝐼2 through the reaction

2𝑀𝑛𝑂2 + 2𝐼 − + 4𝐻 + → 𝑀𝑛2+ + 𝐼2 + 2𝐻2 𝑂 (2)

Next the sample is titrated with standardized sodium thiosulphate(𝑁𝑎2 𝑆2 𝑂3 ∙ 5𝐻2 𝑂)In
the titration process, the 𝐼2 is convert back to 𝐼 − . The amount of iodine molecule present
is directly related to the amount of 𝑂2 original sample.
 2𝑆2 𝑂3 2− + 𝐼2 → 𝑆4 𝑂6 2− + 2𝐼 − (3)

The endpoint of this titration can be determined by potentiometer or by using calorimetric

indicator (starch), which turn deep blue colour to clear.

The DO concentration can determine using the equation: (4)

𝐼2 𝑀𝑛𝑂 𝑂2 32 𝑔 𝑂2 1000 𝑚𝑔
[(𝐿 𝑆2 𝑂3 2− )(𝑀 𝑆2 𝑂3 2− ) ( ) ( 𝐼 2 ) (2 𝑀𝑛𝑂 )( ) ( 𝑔 𝑂 )]
2 𝑆2 𝑂3 2− 2 2 𝑚𝑜𝑙 2
𝑚𝑔𝑂2 /𝐿 =
𝐿 𝑠𝑎𝑚𝑝𝑙𝑒

Nowadays, electrode method is faster, easy operation and less disturbance compare to
Iodometric method. It takes just a few minutes to measure the concentration of DO inside
the water sample.

3.0 MATERIALS AND APPARATUS

Chemicals/Solutions

i. Manganese sulphate
ii. Alkali-iodide-azide reagent
iii. Concentrated 𝐻2 𝑆𝑂4
iv. Starch solution
v. Sodium thiosulphate titrant, 0.0250 M
vi. Water samples (tap water and pond water)
Equipment

i. Conical flasks
ii. Graduated cylinder
iii. 25mL Burette
iv. Burette stand
v. Funnel
vi. Sample bottles with stopper
vii. Three 1.00 mL pipettes

4.0 PROCEDURE

i. 300mL water sample is filled carefully into Biological Oxygen Demand (BOD)
stoppered bottle and the air bubbles are excluded by stopper the bottle.

ii. 1mL of 𝑀𝑛𝑆04 solution is added to the sample bottle, and then followed by 1mL of
alkali-iodide-azide reagent. The reagents are added by inserting the pipette just above
the surface of the liquid and add slowly to prevent the air bubbles from introducing
into the sample.
 iii. The sample bottle is stoppered to exclude air bubbles. Sample is mixed by turning it
upside down several times brownish-orange cloud of floc will appear and let it settle
to bottom.

iv. 1mL of concentrated 𝐻2 𝑆𝑂4 is added. Stopper carefully and shacked until the floc
dissolved.

v. Filled the burette with thiosulphate solution and the initial value is recorded.

vi. 200mL of the water sample is measured and poured into the conical flask. Titrated the
water sample with sodium thiosulphate solution. The sample is first titrated to pale
straw colour and few drops of starch indicator is added (blue colour water form) then
it is titrated to a clear endpoint.

vii. The final reading of burette is recorded in data table.

viii. All steps are repeated for another water sample from a different source.

5.0 RESULTS

Table 1: Titration water samples with thiosulphate solution to clear endpoint

Burette initial Burette final Volume of

Sample
reading, i (mL) reading, f (mL) titrant, f-I (mL)

A (Tap Water) 0 8.5 8.5

B (Pond Water) 10 12 2.0

5.1 CALCULATIONS

𝑪𝒐𝒏𝒄𝒆𝒏𝒕𝒓𝒂𝒕𝒊𝒐𝒏 𝒊𝒐𝒏 𝒐𝒇 𝒅𝒊𝒔𝒔𝒐𝒍𝒗𝒆𝒅 𝒐𝒙𝒚𝒈𝒆𝒏 (𝑫𝑶) 𝒐𝒇 𝒔𝒂𝒎𝒑𝒍𝒆

𝑽𝒐𝒍𝒖𝒎𝒆 𝒐𝒇 𝒕𝒉𝒊𝒐𝒔𝒖𝒍𝒑𝒉𝒂𝒕𝒆 𝒖𝒔𝒆𝒅 (𝑳) × 𝑪𝒐𝒏𝒄𝒆𝒏𝒕𝒓𝒂𝒕𝒊𝒐𝒏 𝒐𝒇 𝒕𝒉𝒆 𝒔𝒐𝒅𝒊𝒖𝒎 𝒕𝒉𝒊𝒐𝒔𝒖𝒍𝒑𝒉𝒂𝒕𝒆 (𝑴) × 𝟖 × 𝟏𝟎𝟑
=
𝑽𝒐𝒍𝒖𝒎𝒆 𝒐𝒇 𝒔𝒂𝒎𝒑𝒍𝒆 (𝑳)

Sample A

8.5 × 10−3 × 0.0250 × 8 × 103

𝐶𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛 𝑖𝑜𝑛 𝑜𝑓 (𝐷𝑂) 𝑖𝑛 𝑡𝑎𝑝 𝑤𝑎𝑡𝑒𝑟 =
200 × 10−3

= 8.5 𝑚𝑔/𝐿
Sample B

2 × 10−3 × 0.0250 × 8 × 103

𝐶𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛 𝑖𝑜𝑛 𝑜𝑓 (𝐷𝑂) 𝑖𝑛 𝑝𝑜𝑛𝑑 𝑤𝑎𝑡𝑒𝑟 =
200 × 10−3

= 2.0 𝑚𝑔/𝐿

5.2 DISSCUSION

Dissolve oxygen in water can be determine by using Winkler Method. In this experiment
water samples are titrated by thiosulphate solution and the volume of titration is recorded.
DO concentration can be determine using formula (4).

From the experiment conducted, the concentration DO for tap water is 8.5 mg/L and for pond
water is 2.0 mg/L. Theoretically the concentration DO for pond water is higher than tap water
because the concentration of organic waste is higher than the tap water. The higher the
organic waste cause, the lower the oxygen dissolve in the water. In our experiment results the
concentration DO of the pond water is lower due to the mechanical problem of the pump of
the pond destruct causes low organic waste flow into the pond.

There are also a lot of others factor that affecting the concentration of DO of water. Several
modifications of the Winkler Method have been developed to overcome interferences like
azide modification.

6.0 EXERCISE

i. Why is dissolved oxygen important in aquatic environment?

The dissolved oxygen level of water is an important indicator of the health of the aquatic
ecosystem. Aquatic life need dissolved oxygen to respire. Microbes such as bacteria and
fungi also use DO to decompose organic material at the bottom of a body of water. The
process is important to contribute the nutrient in food chain and web for the aquatic living
things.

ii. Give three factors affecting the concentration of dissolved oxygen. Discuss each
factor.
There are a lot of factors affecting the concentration of dissolved oxygen (DO) such as
Temperature of water, velocity of water, atmospheric pressure, photosynthesis process of
aquatic plant and organic waste.

Temperature. The concentration of (DO) in cold water holds more than warm water. The
solubility of oxygen decreases as water temperature increases. This is because oxygen is less
soluble in warm water than in cool water. Cooler water is denser than warmer water. Cold
water is dense and compact compare to warm water.

Velocity of Water. Oxygen is very reactive and sensitive easily enters water more quickly
when water mixes with air. More oxygen will mix with water when billow.
Organic Waste. Human-Related Organic Waste which include sewage, urban as well as
discharge from chemical processing industries, meatpacking plants, and dairy waste products.
Organic waste (waste that was once living plants or animals) makes oxygen levels drop
because bacteria that feed on organic wastes consume oxygen. Natural Sources of Organic
Waste. Natural organic waste is created from decaying plant and animal material in a stream,
or when leaves fall into a river.

iii. What is the purpose of the sodium azide modification to the Winkler titration
procedure?
Azide is added to suppress interference from any nitrite present which would react with the
iodide.

You titrate 200mL of sample with 0.025M thiosulphate and the titration takes 8.65mL
of thiosulphate to reach the end point. What is the DO content of the sample?

8.65 × 10−3 × 0.0250 × 8 × 103

𝐶𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛 𝑖𝑜𝑛 𝑜𝑓 (𝐷𝑂) 𝑖𝑛 𝑠𝑎𝑚𝑝𝑙𝑒 =
200 × 10−3
= 8.65 𝑚𝑔/𝐿

7.0 CONCLUSION

In this experiment we learn that the important of dissolve oxygen for the aquatic life to
survive and how it take an important role for our society. The oxygen dissolves by diffusion
from the surrounding into the water and photosynthesis of the green plant in water.

Dissolved oxygen level is important for the aquaculture. The dissolved oxygen level of water
is an important indicator of the health of the aquatic ecosystem. Productive waters contain
adequate amounts of DO to support life forms and maintain that oxygen levels in a stable,
healthy range. Dissolve oxygen level also used in the processing of waste water treatment.

8.0 REFERENCE

a. http://www.fondriest.com/environmental-measurements/parameters/water-
quality/dissolved-oxygen/, Doug Nguyen, 1999-07-18
b. https://www.lenntech.com/why_the_oxygen_dissolved_is_important.htm, enntech
Water &Luchtbehandeling Holding B.V., 2000-03-01
c. http://www.oocities.org/rivertalkhome/DissolvedOxygen.html, Pasch F., 2010-09-14