ABSTRACT / SUMMARY

An experiment called Biochemical Oxygen Demand (BOD) test

is conducted in order to determine the amount of dissolved oxygen
contains in water sample collected from Tasik Seksyen 7, Shah
Alam. The method comprises mainly three stages, which are
collection of water sample, stabilization of water sample with
manganous sulfate and alkali iodide – azide reagent powder pillows,
and titration of the sample with sodium thiosulfate cartridge plus
addition of starch solution. Besides that, this experiment also
discusses the importance of dissolved oxygen for aquatic ecosystem,
the reasons of reduction level of dissolved oxygen which is
temperature, pressure, salinity and elevation of water bodies as well
as the circumstances of the depletion of concentration of dissolved
oxygen to the survival of aquatic ecosystem. Based on the
experiment result, the amount of dissolved oxygen in the water
sample is 6.62 mg/L. The experiment is completed and conducted
successfully.

INTRODUCTION

Oxygen is indeed a well known fact as the air that both humans
and animals need for respiratory system. Dissolved oxygen is oxygen
in the form of water which is freely available to aquatic plants and
animals including fish, invertebrates, plants and aerobic bacteria.
The existence of dissolved oxygen is by the oxygen that are being
transferred from the atmospheres into water surfaces along with the
oxygen produced by aquatic plants, algae and phytoplankton as a by-
product of photosynthesis. Once the oxygen is dissolved in water, it
diffuses very slowly throughout a water body. The distribution is slow
due to the dependency on the movement of aerated water, which is
the water that is over flown with carbon dioxide, by turbulence and
currents, water flow as well as thermal upwelling.

The level of dissolved oxygen is an indicator for water body’s

ability to support desirable aquatic life. The species and the stage of
life of a species are the variables that determine the amount of
oxygen. As for growth and activity, the required amount are between
range of five to six parts per million (5 – 6 ppm). In addition,
dissolved oxygen levels below three parts per million (3 ppm) is
dicey and means harm to most aquatic organisms. It gets even worse
if the amount of dissolved oxygen is below two parts per million (2
ppm) as the aquatic livings especially fish may not survive. Oxygen
is particularly sensitive and hardly available as over period of time,
its existence is depending upon the temperature, pressure, salinity
effect, other chemicals present in water and biological processes.
Temperature indeed plays a major role in influencing the amount of
dissolved oxygen in water. Water at a temperature of 31 °C only
contains half of dissolved oxygen contents in cold atmosphere at
1°C. This shows that cold water has the ability to dissolve oxygen
greater than in warm water.

Oxygen from the atmosphere is transferred into surface water

at the point of contact where it interfaces with the air. Points of
contact are elevated corresponding to the wind behaviour through a
process called physical aeration. The oxygen is also comprises of
the by-product of photosynthesis process. Likewise, the levels of
dissolved oxygen are increased by rooted and floating aquatic plants
through this process. The process of producing oxygen usually can
occur near the surface of plants or in shallow water where light is
permissible due to the fact that plants depends on availability of
light to live and survive. Apart from that, oxygen is considered to be
harmful for consumption at night or on cloudy days where oxygen
producing via photosynthesis process could not occur because of
lacking of sunlight. Hence, nearly all aquatic plants organisms use
the dissolved oxygen to move and sustain in life. This process,
called respiration can in turn reduce the concentration of oxygen, in
this context, to the risky low levels.

There is however a lot more factors that contributes to the

depleting of dissolved oxygen in water. One of it is due to the
presence of organic wastes. The decaying process of dead
organisms or organic wastes consumes oxygen and is often
concentrated in summer. This is because the aquatic animals require
more oxygen for higher metabolism. Depletions of dissolved oxygen
level can cause major shifts in the kinds of aquatic livings. This
situation may cause catastrophe or further difficulties for human
beings especially, as aquatic livings are one of our source of food.

The decrease in level of dissolved oxygen is also related to

pressure. As the barometric pressure of atmosphere decreases, the
concentration of dissolved oxygen also decreases. This occurs due
to the increase in the altitude or elevation of water body. As an
example, consider the barometric pressure in the mountains, which
apparently is less than the barometric pressure in the oceans. Thus,
at higher altitudes, water holds less dissolved oxygen compared to
the condition at lower altitudes.

OBJECTIVE

The main objective of this experiment is to determine the

amount of dissolved oxygen in water sample. Besides that, a few
minor objectives are also considered, which include relating the
concept of oxygen with levels of dissolved oxygen in the aquatic
ecosystem, discussing the importance of measurement dissolved
oxygen as water quality as well as studying the reasons and
circumstances or effect corresponding to the change in level of
dissolved oxygen.

THEORY

The concentrations of dissolved oxygen may vary drastically

depending upon depth and distance of the water bodies from shore.
There are three different methods of measuring the level of
dissolved oxygen. The first one is the Winkler titration method,
where it requires lots of use of strong chemicals. Thus, it is not
often used for citizen monitoring programs. The second method is by
basically using a dissolved oxygen probe and meter. Nevertheless,
the probes are easily ruined as the membrane deteriorates, besides
the trapping of air bubbles under the membrane and contamination of
the sensing element. It is also not easy to assure whether the probe
is functioning properly. The third method is by using dissolved
oxygen field kits. The kits are user-friendly, convenient and come
with simple instructions. Different kit manufacturer however has
different instruction of using it. This test also requires short period
of time. However, this field kits method mostly does not meet the
requirements for precision and accuracy needed for professional
quality data.

The decrease in level of dissolved oxygen is related to

pressure. As the barometric pressure of atmosphere decreases, the
concentration of dissolved oxygen also decreases. This occurs due
to the increase in the altitude or elevation of water body. As an
example, consider the barometric pressure in the mountains, which
apparently is less than the barometric pressure in the oceans. Thus,
at higher altitudes, water holds less dissolved oxygen compared to
the condition at lower altitudes.

The experiment conducted is also known as biochemical oxygen

demand (BOD) test, and is no different with Winkler method. It is
generally a technique used in order to determine and measure the
level of dissolved oxygen in water bodies. Dissolved oxygen is
important to be measured as it indicates the condition or productivity
of the water source as high concentration of dissolved oxygen gives
a corresponding results that the water is of high productivity and
little pollutant.
 This experiment uses titration method where water sample is
filled completely in a specific bottle called Biochemical Oxygen
Demand (BOD) bottle. There must be no air bubbles contain in the
water sample and the dissolved oxygen content is then added with a
series of reagents that form an acid compound, which is then titrated
with neutralizing compound by considering any colour change.

This test technically involves three main stages; collection of

water sample, stabilization of the water sample, as well as titration
the water to determine the value of dissolved oxygen. The contact
between the water sample and the atmosphere will not affect much of
the results, so it is not necessary to perform the procedure
immediately after the water sample is collected.

However, this test does not give any information on the amount
of the dissolved oxygen the water is capable of holding at the
temperature of when it is tested. When water holds all the possible
dissolved oxygen at a given temperature, it is considered to contain
hundred percents (100%) of saturated oxygen.

EXPERIMENTAL PROCEDURE

1.) A water sample is collected in a clean 300-mL BOD Bottle.

2.) The contents of the Manganous Sulfate Powder Pillow and one
Alkaline Iodide-Azide Reagent Powder Pillow are added.
3.) The stopper is inserted immediately so air is not trapped in the
bottle. The stopper is inverted several times to mix.

4.) The floc in the solution is waited to settle. The bottle is inverted
again several times and the floc is waited to settle.

5.) The stopper is removed and the content of one Sulfamic Acid
Powder Pillow is added. The stopper is replaced without trapping air
in the bottle and inverted the prepared sample several times to mix.

6.) A sample volume and Sodium Thiosulfate Titration Cartridge are

selected corresponding to the expected dissolved oxygen (D.O)
concentration.

7.) A clean delivery tube is inserted into the titration cartridge. The
cartridge is attached to the titrator body.

8.) The delivery knob is turned to eject a few drops of titrant. The
counter is reset to zero and the tip is wiped.

9.) A graduated cylinder is used to measure the sample volume from

Table1. The sample is transferred into a 250-mL Erlenmeyer flask.

10.) The delivery tube tip is placed into the solution and the flask is
swirled while titrated with sodium thiosulfate to a pale yellow colour.

11.) Two 1-mL droppers of Starch Indicator Solution are added and
swirled to mix.

12.) The titration is continued to a colourless end point. The number

of digits required is recorded.

13.) The value is calculated using the equation:-

mg/L Dissolved Oxygen = Digits Required x Digit Multiplier.

 APPARATUS / REAGENTS

~ Manganous Sulfate Powder Pillow, Alkaline Iodide-Azide Reagent

Powder Pillow, Sulfamic Acid Powder Pillow, Sodium Thiosulfate
Titration Cartridge, 0.2000N, Sodium Thiosulfate Titration Cartridge,
2.0000N, Starch Indicator Solution, 300-mL BOD bottle, graduated
cylinder, delivery tube, conical flask, and digital titrator.

RESULT

Range Sample Titration cartridge Digit Digit Result

(mg/L) volume (mL) (N Na2S2O3) multiplier required (mg/L O2)
100 0.200 0.02 331 6.62
Observation
The floc appears soon after MgSO 4 powder pillow and alkaline
2 - 10
iodide-azide reagent powder pillow is mixed, and disappears.
Oxygen is present as the solution turn orange-brown. Pale yellow
colour is formed after sulfamic acid powder pillow is mixed.

SAMPLE CALCULATION
The total number of millilitres of titrant used equals the total
dissolved oxygen in the sample in mg/L. In order to calculate the
content of dissolved oxygen in water sample the following expression
is used;

Digit required x Digit multiplier = mg/L of dissolved oxygen

311 x 0.02 = 6.62 mg/L

DISCUSSION

From the experiment, a floc is observed to appear in the water

sample soon after the addition of Manganous Sulfate and Alkaline
Iodide – Azide powder pillows are added. The floc is in orange-brown
colour, indicating the presence of dissolved oxygen. The stabilized
water sample is then titrated with sodium thiosulfate and turns pale
yellow. When the starch solution is added, the solution turns
colourless, indicates the endpoint of the titration process. The
amount of dissolved oxygen is calculated to be a value of 6.62 mg/L,
which falls in the range required for 0.200 N of sodium thiosulfate,
which is between 2 to 10 mg/L.

Nevertheless, there must have been some errors made during

the experiment. In my opinion, the value of dissolved oxygen
obtained from the water sample collected from ‘Tasik Seksyen 7,
Shah Alam’ is not the actual value, due to the presence of air
bubbles inside the water sample when it is filled in the BOD bottle.
There is also little air bubbles present in the delivery tube, which
results in the inaccuracy of the amount of dissolved oxygen. Besides
that, the water sample is not collected at near the middle of the lake
or at arm’s length below the water surface. The starch solution
added may also not be of accurately 1 millilitre for each drop, which
also adds to reasons of my perspective that the amount of dissolved
oxygen obtained is not the precise value.

CONCLUSION

The amount of dissolved oxygen in the water sample is 6.62

mg/L, hence the water sample is considered to have high
concentration of dissolved oxygen and therefore, high productivity
and less polluted.

RECOMMENDATION

There are a few recommendations and precautions that have to

be taken account in this experiment.

Firstly, the water sample is best collected at near the middle of

the water stream and at least a few inches or at arm’s length below
the water surface. This is to ensure that the water is collected right
near the presence of aquatic organisms.

Secondly, the difference between the time the water sample is

collected and the time the experiment is conducted should not be too
long, as the temperature and barometric pressure of atmosphere
during the collection of water sample will change over a time period.

Thirdly, in order to avoid or eliminate the presence of air

bubbles inside the water sample, it is a must to cap the BOD bottle
immediately after the water sample is filled in. If air bubbles are
detected, remove the water sample and refill with another sample.
The water sample should be allowed to overflow the BOD bottle at
least 2 minutes or until the water in the bottle has replaced itself two
or three times. The bottle may also be inverted or turned upside
down to observe any bubble movement. Besides that, the bottle may
be squirted with distilled water along the stopper, capped with
aluminium foil and rubber band.

The reagents used in this experiment are mostly strong

chemicals and hazardous to one’s health. Therefore, cautions are
required when handling these reagents. Gloves and goggles are used
in order to prevent any unexpected contact with the reagents.

Besides that, the titration should be conducted slowly to allow

the solution to react and mix with each other to reach their
equilibrium state along the swirling and stirring of the solution. When
adding the starch solution, it is best to hold the flask up to a white
sheet of paper to observe the disappearance and absence of blue
colour.

REFERENCES

1. http://www.fivecreeks.org/monitor/do.html
2. http://serc.carleton.edu/microbelife/research_methods/environ_
sampling/oxygen.htm
3. http://www.ecy.wa.gov/programs/wq/plants/management/joysmanual/
4oxygen.html

APPENDICES
 Digital titrator BOD bottle

Titration cartridge
Hach DREL/2400 Basic Water Quality Laboratory Delivery tube