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 3.78 mg/L and 4.00 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 acceptable to support the population in the lake where water sample is
taken.

2. Introduction.
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.
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 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 wastewater 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 wastewater 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

water
CO2

oxygen
+

H 2O

carbon-rich food
O2

C6H12O6

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 wastewater 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:
a) To determine the DO level in a series of water samples.
b) Ascertain whether the water samples are comply with Malaysian Water
Standards or not.

5. Apparatus.
60mL of water sample (lake water), 300mL of water sample (lake water), Manganous
Sulphate Powder Pillow, Alkaline Iodide Azide Reagent Powder Pillow, Sulfamic Acid
Powder Pillow, Sodium Thiosulphate solution, 2mL of starch solution.
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)
Firstly, water sample was collected and poured in the clean 300 ml BOD Bottle.
Manganous Sulfate Powder Pillow and one Alkaline Iodide Axide Reagent Powder
Pillow were added. Stopper was inserted immediately so that there was no air
trapped in the bottle. The bottle was inverted a few times so that it mixed well.bottle
was left to make sure the floc was settled. Stopper was removed and one Sulfamic
Acid Powder Pillow was added. Stopper was inserted without trapping air and
inverted a few times. The floc will disappear and the solution turns yellow.100 ml of
solution was measured and transferred into the conical flask. Clean delivery tube
was inserted into the titration cartridge contains 0.2 M Sodium Thiosulfate Titration.
The prepared solution was titrated by with 0.2 M Sodium Thiosulfate until yellow
solution turns into pale yellow colour. Starch Indicator Solution was inserted into the
sample and the mixture was stirred well. Titration was continued until the pale yellow
solution turns into colourless. Number of digits that appear was recorded and
calculation for oxygen dissolved was calculated.

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

Method)
Firstly, water sample was collected and poured in the clean 60 ml BOD Bottle. One
Dissolved Oxygen 1 Reagent Powder Pillow and one Disslved Oxygen 2 Reagent
Powder Pillow were added. Stopper was inserted immediately so that there was no
air trapped in the bottle. The bottle was inverted a few times so that it mixed
well.bottle was left to make sure the floc was settled. Stopper was removed and one
Dissolved Oxygen 3 Powder Pillow was added. Stopper was inserted without
trapping air and inverted a few times. The floc will disappear and the solution turns
yellow.20 ml of solution was measured and transferred into the conical flask. Clean
delivery tube was inserted into the titration cartridge contains 0.2 M Sodium
Thiosulfate Titration. The prepared solution was titrated by with 0.2 M Sodium
Thiosulfate until yellow solution turns into pale yellow colour. Starch Indicator
Solution was inserted into the sample and the mixture was stirred well. Titration was
continued until the pale yellow solution turns into colourless. Number of digits that
appear was recorded and calculation for oxygen dissolved was calculated.

7. Results.
Table 1
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

8. Calculations.

Titration using a 300 mL BOD bottle:

Orange

Pale yellow = 179

Blue

Colourless = 189

Digit required x Digit multiplier

189

0.02

3.78 mg/L

Titration using a 60 mL BOD bottle:

Orange

Colourless = 40

Digit required x Digit multiplier

40

0.1

4.00 mg/L

Catalog No.

Digit
Multiplier

22675-01
22675-01
14401-01

0.01
0.02
0.1

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 3.78 mg/L and 4.00 mg/L respectively. From the
data, the DO level for the water sample taken is acceptable to support the population
in the lake 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 interferences, 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 3.78 mg/L and 4.00
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 acceptable to
support the population in the lake where water sample is taken.

11. Recommendations.
There are several errors during experiments. Firstly, the apparatus such as beakers
are not being rinsed with distilled water before use and some of apparatus such as
beaker not completely dry and clean and give effect to solution. So, the beaker,
pipette, burette and the flask should be washed properly with distilled water. At the
same time, make sure the apparatus such as beaker is completely dry and clean.
Next, repeat the experiment a few times to get more accurate result. At all stages,
precaution must be taken to ensure that oxygen is neither introduced nor lost from
the water samples.

12. References.

i)

http://kmam.moh.gov.my/public-user/drinking-water-quality-standard.html
[ Retrieved on 23/11/2014 ]

ii)

http://water.epa.gov/type/rsl/monitoring/vms52.cfm
[ Retrieved on 23/11/201 ]

iii)

http://www.water-research.net/index.php/dissovled-oxygen-in-water
[ Retrieved on 23/11/2014 ]

iv)

http://nitttrc.ac.in/Four%20quadrant/eel/Quadrant%20%201/exp10_pdf.pdf [ Retrieved on 21/11/2014 ]

v)

http://water.me.vccs.edu/courses/env211/lab14.htm
[ Retrieved on 21/11/2014 ]

Group members :
1. Nur Syahira Hazwani Binti Hamran
2014276948
2. Adibah Binti Abd Aziz
2014854258
3. Mafazrina Asyikin Binti Naspu
2014807024
4. Nur Amira Binti Khiruddin
2014670704
5. Nik Nurilya Hazlin Binti Nik Lah
2014807764