1.

Abstract

The purpose of this experiment is to determine the DO level in a series of water

samples and as certain whether they comply with Malaysian Water Standards or not. The
Winkler-method is a technique that used to determine the dissolved oxygen in water samples.
Firstly, Winkler-method is a technique used to determine the dissolved oxygen in water
samples. The value of dissolved oxygen tested for each bottle is 6.85 mg/L and 13.3 mg/L for
300 mL and 60 mL BOD bottles respectively. In conclusion, based on Malaysian Water
Standards, the DO level for the water sample taken is not acceptable to support the population
in the lake where water sample is taken.

2.

Introduction

Dissolved oxygen (DO) is naturally present in lakes and rivers to maintain the healthy,
quality of aquatic environment and to sustain aquatic life. DO is almost important for
metabolic processes of almost every organism.The term of oxygen saturation or dissolved
oxygen (DO) is used to measure the relative amount of oxygen dissolved in a medium or a
unit volume of water.
The dissolved oxygen content of water is infected by the source, temperature of water
sample, chemical or biological process taking places and environment. The temperature will
determine the saturation point oxygen in water. The colder the water, it will hold more
oxygen and vice versa. Water that is flowing or running through the stream typically will
have higher concentration of DO than stagnant water. It is because the flowing water has a
greater contact of surface area with air that contains oxygen than stay still water. Winds are
that in contact with water will allow oxygen to mix with it. Windless night will cause oxygen
depletion in aquatic life. Other than that, aquatic plants also will release oxygen into water
during photosynthesis during the day.
Present of oxygen in the water is a good thing. Microorganism include fish and other
aquatic organism in water will use up the oxygen just like we use up oxygen in the air.
During the night, aquatic plants will take in oxygen rather than releasing it like they do
during the day. The intake of oxygen eventually will decrease the DO levels.
DO concentration level in water is important because it determines which organisms
are being able to live in water, and also waste water treatment plants. If the concentration of
DO level deplete, it will encourage the microbial reduction of nitrate to nitrite, sulphate to
sulphide and increase the concentration of ferrous iron in solution. Waste water treatment are
depends on bacteria to breakdown the organic compounds. If amount of dissolved oxygen in
waste water becomes low, the bacteria will die and septic conditions will occurs that result
unpleasant smell and increase the killing of organism. When it reach one level, most of waste
will break down, concentration of DO level will rises as the oxygen in river had recovers
from air and aquatic plant by photosynthesis. Thus, DO is only a basis test but an important
parameter to evaluate organic pollution of waste
For information, water present in lakes, river or stream, the dissolved oxygen is about
8 parts per million. Minimum DO level of 4 or 5 mg/L is good for aquatic to survive.

3.

Theory

Dissolved oxygen analysis measured the amount of oxygen dissolved in an aqueous

solution. Oxygen gets into the water by diffusion from the surrounding air by rapid
movement (aeration) and also as a waste product of photosynthesis process. A simple formula
of photosynthesis which is in the condition of the presence of light and chlorophyll is shown
below:
Carbon dioxide +

CO

water
2

oxygen

carbon-rich food

H 2 O 02 C 6 H 12 O6

The concentration of dissolved oxygen is usually expressed in term of milligram of

oxygen per liter of water (mg/L) or in parts per million (ppm). Some water compare
calculated oxygen content with observed concentration and report percent saturation (% sat).
Oxygen enters the water by direct absorption from the atmosphere or by plant
photosynthesis. The oxygen is used by plants and animals for respiration and by the aerobic
bacteria which consume oxygen during the process of decomposition. When organic matter
such as animal waste or improperly treated waste water enters a body of water, algae growth
increases and the dissolved oxygen levels decrease as the plant material dies off and is
decomposed through the action of the aerobic bacteria.
Decreases in the dissolved oxygen levels can cause changes in the types and numbers
of aquatic macro invertebrates which live in a water ecosystem. Species which cannot tolerate
decreases in dissolved oxygen levels include mayfly nymphs, stonefly nymphs,caddisfly
larvae and beetle larvae.As the dissolved oxygen level decrease, these pollution-intolerant
organisms are replaced by the pollution-tolerant worms and fly larvae.
Dissolved oxygen levels change and vary according to the time of day, the weather and
the temperature. If yearly comparisons are made on dissolved oxygen levels, they should be
done at the same time of day, during the same season and on day with a temperature variation
of only 10 degree Celsius from the previous reading.

Dissolved oxygen analysis can be used to determine:

1. The health or cleanliness of a lake or stream
2. The amount and type of biomass a freshwater system can support
3. The amount of decomposition occurring in the lake or stream
Dissolved oxygen can be calculated by:
Digits required Digits multiplier = _____mg/l

Dissolved oxygen is widely used in water quality studies and daily operations of
water reclamation facilities. An Dissolved Oxygen 1 Powder Pillow and Dissolved Oxygen 2
Powder Pillow are added to a water sample causing a white floc of is formed. This precipitate
is then oxidized by the Dissolved Oxygen 3 Powder Pillow which added in the water sample
into a brown-yellow floc. The amount of dissolved oxygn is directly proportional to the
titration a sodium thiosulphate solution.
There are two common methods for measuring DO. The firsts and most reliable is the
Azide-Winkler titration method, against which the others are compared to test for accuracy.
However, this method also requires the most training and the use of some strong chemicals.
For these reasons, it is not often used in citizen monitoring programs. The second and
probably most common method is the use of a DO probe and meter.

4.

Objectives

The objectives of conducting this biochemical oxygen demand (BOD) experiment are:
i. To determine the DO level in a series of water samples.
ii. Ascertain whether the water samples are comply with Malaysian Water
Standards or not.

5. Methodology

5.1 Material
60mL of water sample (Delima Colleges Drain water)
300mL of water sample (Delima Colleges Drain water)
Manganous Sulphate Powder Pillow

 Alkaline Iodide Azide

Reagent Powder Pillow
Sulfamic Acid Powder Pillow
Sodium Thiosulphate solution
2 mL of starch solution.

5.2 Apparatus
60mL of BOD bottle
300mL of BOD bottle
50mL flask
Titration Catridge.

6.

Procedures

6.1 Oxygen dissolved using a 300 ml BOD Bottle ( Axide Modification of Winkler Method)
i.

Water sample was collected and poured in the clean 300 ml BOD Bottle.

ii. Manganous Sulfate Powder Pillow and one Alkaline Iodide Axide Reagent Powder
Pillow were added.
iii. Stopper was inserted immediately so that there was no air trapped in the bottle.
iv.

The bottle was inverted a few times so that it mixed well. Bottle was left to make
sure the floc was settled.

v.

Stopper was removed and one Sulfamic Acid Powder Pillow was added.

vi. Stopper was inserted without trapping air and inverted a few times. The floc will
disappear and the solution turns yellow.
vii. 200 ml of solution was measured and transferred into the conical flask.
viii. Clean delivery tube was inserted into the titration cartridge contains 0.2 M Sodium
Thiosulfate Titration.
ix. The prepared solution was titrated by with 0.2 M Sodium Thiosulfate until yellow
solution turns into pale yellow colour.
x.

Starch Indicator Solution was inserted into the sample and the mixture was stirred
well.

xi. Titration was continued until the pale yellow solution turns into colourless.
xii. Number of digits that appear was recorded and calculation for oxygen dissolved was
calculated.
xiii.

6.2 Oxygen dissolved using a 60 ml BOD Bottle ( Axide Modification of Winkler Method)

i.

Water sample was collected and poured in the clean 60 ml BOD Bottle.

ii. One Dissolved Oxygen 1 Reagent Powder Pillow and one Dissolved Oxygen 2
Reagent Powder Pillow were added.
iii. Stopper was inserted immediately so that there was no air trapped in the bottle.
iv. The bottle was inverted a few times so that it mixed well. Bottle was left to make
sure the floc was settled.
v.

Stopper was removed and one Dissolved Oxygen 3 Powder Pillow was added.

vi. Stopper was inserted without trapping air and inverted a few times. The floc will
disappear and the solution turns yellow.
vii. 20 ml of solution was measured and transferred into the conical flask.
viii. Clean delivery tube was inserted into the titration cartridge contains 0.2 M Sodium
Thiosulfate Titration.
ix. The prepared solution was titrated by with 0.2 M Sodium Thiosulfate until yellow
solution turns into pale yellow colour.
x.

Starch Indicator Solution was inserted into the sample and the mixture was stirred
well.

xi. Titration was continued until the pale yellow solution turns into colourless.
xii. Number of digits that appear was recorded and calculation for oxygen dissolved was
calculated.

7.

Results

Range (mg/L
D.O)

Sample volume
(mL)

1-5
2-10
>10

200
100
200

Titration
Catridge (N
Na2S2O3)
0.200
0.200
2.000

Catalog No.

Digit Multiplier

22675-01
22675-01
14401-01

0.01
0.02
0.1

Table 7.1: Digit multiplier with volume of water sample taken.

8.

Calculations

8.1 Titration using a 300 mL BOD bottle:

Sample volume = 200 mL
Titration catridge =

0.2 N Na S O
2

Digital required = 685

Digit required x Digit multiplier
685

0.01

6.85 mg/L

8.2 Titration using a 60 mL BOD bottle:

Sample volume = 20 mL
Titration catridge = 0.2 N
Digital required = 133

Na S O
2

Digit required x Digit multiplier

133

0.1

13.3 mg/L

9. Discussions
Dissolved oxygen analysis measures the amount of gaseous oxygen dissolved in an
aqueous solution. Oxygen gets into water by diffusion from the surrounding air, by aeration
(rapid movement), and as a waste product of photosynthesis. As dissolved oxygen levels in
water drop below 5.0 mg/L, aquatic life is put under stress. The lower the concentration, the
greater the stress. Oxygen levels that remain below 1-2 mg/L for a few hours can result in
large fish kills. In the experiment, the sample is collected in 300 mL and 60 mL BOD bottles
and the value of dissolved oxygen tested for each bottle is 6.85 mg/L and 13.3 mg/L
respectively. From the data, the DO level for the water sample taken is not acceptable to
support the population in the drain where water sample is taken.
There are several errors that may lead to inaccurate data. Firstly, the water sample must
be taken before afternoon as DO level will raise rapidly during afternoon because a lot of
oxygen is used up by plants and animals in the lake. Next, the sample must be tested in-situ to
avoid loss of oxygen to the surroundings. The sample must be kept from atmospheric oxygen
to avoid inaccurate data. Finally, in the determination of DO level by the Winkler method,
there are various materials which can cause interference, including iron salts, organic matter,
excessive suspended matter, sulfide, sulfur dioxide, residual chlorine, chromium, cyanide,
and certain oxidizing and reducing agents that may influence the readings taken.

10.

Conclusion

The value of dissolved oxygen tested for each bottle is 6.85 mg/L and 13.3 mg/L for
300 mL and 60 mL BOD bottles respectively. In conclusion, based on Malaysian Water
Standards, the DO level for the water sample taken is not acceptable to support the population
in the drain where water sample is taken.

11.

Recommendations

There are several errors during experiments. Thus, few recommendations has been made;
i.

The apparatus used should be washed properly with distilled water and ensure it is
completely dry to avoid any effect on the solution.

ii. The experiment should be repeated at least twice to obtain the more accurate and
average results.
iii. At all stages, precaution must be taken to ensure that oxygen is neither
introduced nor lost from the water samples.

12.

References

i)

http://water.epa.gov/type/rsl/monitoring/vms52.cfm
[ Retrieved on 05/11/2015 ]

ii)

http://nitttrc.ac.in/Four%20quadrant/eel/Quadrant%20-%201/exp10_pdf.pdf
[ Retrieved on 08/11/2015 ]

iii)

http://www.water-research.net/index.php/dissovled-oxygen-in-water
[ Retrieved on 08/11/2015 ]

iv)

http://kmam.moh.gov.my/public-user/drinking-water-quality-standard.html
[ Retrieved on 10/11/2015 ]

v)

http://water.me.vccs.edu/courses/env211/lab14.htm
[ Retrieved on 11/11/2015 ]