TABLE OF CONTENT

CONTENTS Page number

1.0: ABSTRACT 2

2.0: INTRODUCTION 3

3.0: AIMS 6

4.0: THEORY 7

5.0: APPARATUS 9

6.0: METHODOLOGY 10

7.0: RESULT 11

8.0: SAMPLE CALCULATION 12

9.0: DISCUSSION 13

10.0: CONCLUSION 16

11.0: RECOMMENDATION 17

12.0: REFERRENCES 18

13.0: APPENDIX 19

-1-
1.0 ABSTRACT

Water quality is a description of chemical, physical and also biological characteristics of water that
should be determined in order to ensure it is safe enough to be use and drink. The aim for this
experiment is to determine the amount of dissolved oxygen in sample water collected in fish pond,
using Winkler Method and then be comply with the Malaysian Standard of Water Quality. The
sample water was added with several reagents which are manganese sulfate, alkali-iodide -azide
reagent and sulfamic acid. After that, the water sample has been titrated with sodium thiosulfate.
The addition of astarch solution was act as an indicator to determine the titration endpoint that will
change the colour of solution from blue to colorless. Through observation, we can see that when we
put manganese sulfate and alkali-iodide-azide reagent into the sample water, the colour of sample
water change to orange colour and there are floc in it with brownish-orange. After the sample water
were added with sulfamic acid powder pillow, the colour change to yellow that means the oxygen is
present. The amount of DO calculated is 6.52 mg/L which is in range of 2-10 mg/L and classified as
Class II in Malaysian Standard of Water Quality.

-2-
2.0 INTRODUCTION

Water is important for our body and the earth. 78% of the earth is covered by water and 70%
of a human’s body contains water. Water also plays an important role in human’s daily life. This is
because, people need water body for detoxification, food digestion process, balancing the body
temperature and for metabolism.

Water quality is a description of chemical, physical and biological characteristics of water.

Water quality has an intrinsic value of life which is often judged on the availibility of pristine water.
Thus, water quality is not only because of the availability of water for various uses and its impact on
public health. When water is contaminated, it may effects the present and future generations. Other
than that, water quality also affected the presence, abundance, diversity and distribution of aquatic
species in surface water too. Even though we can use many method in measuring the quality of
water, still, it is not a simple thing to determined weither that water is good or bad.

There are some parameters for monitoring some activities and the impact to the waterthat
were used by Jabatan Alam Sekitar Malaysia. For an example, the table below shows that some test
needed to be conducted to know the value of DO, pH, conductivity, temperature, Calcium,
Magnesium and Natrium in general.

Table 2.1 Water Quality Parameter and Activities (Alam Sekitar Malaysia Sdn. Bhd, 2004)

Activity Water Quality Parameters

Dissolved oxygen (DO),conductivity,
General water quality temperature, dissolved solid, chloride (Cl),
calcium (Ca), magnesium (Mg), Natrium (Na)
Biochemical Oxygen Demand (BOD), Chemical
Human Activities Oxygen Demand(COD), Phospate (PO4), Oil and
gris.MBAS
Sewage Water Pollution Ammonium, E. Coli
Land projects Suspended Solid (SS)
Agriculture runoff Potassium, Nitrate
Arsenic, Cadmium, Chromium, Lead, Zink,
Industrial waste
Ferrous. Argentums
Table 2.2 Classes of Water Quality and its Uses

Class Uses

-3-
 I Conservation of natural environment water supply 1
- practically no treatment
necessary. Fishery 1- very sensitive aquatic species
IIA Water Supply II - conventional treatment required.
Fishery 11- sensitive aquatic species.
IIB Recreational use with body contact
III Water Supply lll - extensive treatment required.
Fishery 111- common, of economic value, and
tolerant species livestock drinking
IV Irrigation
V None the above

Table 2.3 Proposed National Water Quality Standard for Malaysia

Classes
Parameters Unit
I IIA IIB III IV V
Ammonical-N. mg/l 0.1 0.3 0.3 2.9 2.7 >2.7
BOD mg/l 1 3 3 6 12 >12
COD mg/l 10 25 25 50 100 >100
DO mg/l 7 5-7 5-7 3-5 <3 <1
pH 6.5-8.5 6-9 6-9 5-9 5-9 -
Colour TCU 15 150 150 - - -
Elec. Cond ӿ μmhos/cm 1000 1000 - - 6000 -
Floatables N N N - - -
Odour N N N - - -
Salinity % 0.5 1 - - 2 -
Taste N N N - - -
Tot. Diss. Sol. ӿ Mg/l 500 1000 - - 4000 -
Tot. Susp. Sol. ӿ Mg/l 25 50 50 150 300 >300
Normal ± Normal ±
Temperature ᴼC - - - -
2 2
Turbidity NTU 5 50 50 - - -
F. Coliform Counts/100 5000 5000
10 100 400 -
ӿӿ ml (20000)ᵋ (20000)ᵋ

-4-
 Counts/100
Total Coliform 100 5000 5000 50000 50000 >50000
ml

N = No visible floatable material / debris,

or No objectionable odour,
or No objectionable taste.

* = Related parameters, only one recommended for use

** = Geometric mean

= Maximum not to be exceeded

Dissolved oxygen (DO) is one of the most important parameters in aquatic systems. DO can
be related with the amount of oxygen in the water. DO is an important especially for the equatic lives
for respiration and photosynthesis. There are many thing in this world that needed oxygen. For
example, water from the photosynthesis process of algae, rooted plants and environment needed
oxygen. Other than that, fishes and aquatic animals also use the oxygen everyday. The solubility of
oxygen is increased at low temperature. Thus, cold water have more dissolved oxygen than warm
water. In winter and early spring, when the water temperature is low, the dissolved oxygen
concentration is high. Dissolved oxygen is essential for the support of fish and other aquatic life and
aids in the natural decomposition of organic matter. Waste treatment plant which employ aerobic
digestion must maintain a level of at least 2 ppm dissolved oxygen. This is usually accomplish by
mechanical aeration.

In making drinking water taste better, we needed a high DO level in a community water
supply. However, high DO levels speed up corrosion in water pipe. This is why industries use water
with the least possible amount of dissolved oxygen. The amount of dissolved often determines the
number and types of organism living in that body of water. The amount of oxygen required by
organism refer to the biochemical oxygen demand (BOD). Thus, BOD is defined as total oxygen that
needed by microorganism to decompose and oxidize the organic matter at aerobic condition.

-5-
3.0 OBJECTIVE

The aims of this experiment are:

i. To determine the amount of dissolved oxygen in wastewater.

ii. To comply the result of dissolved oxygen with the Malaysia Standard of Water Quality.

-6-
4.0 THEORY

The Winkler Method is a technique that is used in measuring dissolved oxygen in freshwater systems.
Dissolved oxygen is used as an indicator of the health of a water body. A high concentration of
dissolved oxygen has a high productivity and little pollution. This test is usually performed on-site.
The sample collection and testing method may result in an alteration in oxygen content.

The Winkler Method uses titration to determine dissolved oxygen in the water sample. A
sample bottle is filled completely with water until no bubble left in the bottle. Then, the dissolved
oxygen in the sample bottle is added by a series of reagents that form an acid compound. After that,
it is titrated with a neutralizing compound and results in a color change. The point of color change is
called as endpoint where it equivalent with the dissolved oxygen concentration in the sample water.
Dissolved oxygen analysis is best done in the field, as the sample will be less altered by atmospheric
equilibration.

An excess of manganese salt, iodide and hydroxide ions are added to a water sample and
caused a white precipitate of Mn(OH)2 to form. Then, oxidized the precipitate by the dissolved
oxygen in the water sample into a brown manganese precipitate. After that, a strong acid is added to
acidity of solution. The brown precipitate then convert the iodide ion to Iodine. The amount of
dissolved oxygen is directly proportional to the titration of Iodine with a thiosulfate solution.

Winkle Method has several method to be followed. Firstly, Manganese (II) sulfate is added to
an environmental water sample with a portion of 48% of the total volume. Next, potassium iodide is
added to create a pinkish-brown precipitate at 15% in potassium hydroxide. In the alkaline solution,
dissolved oxygen will oxidize manganese (II) ions to the tetravalent state.

2 Mn(OH)2(s) + O2(aq) → 2 MnO(OH)2(s)

-7-
 MnO(OH)2 appears as a brown precipitate. However, there is some confusion about whether
the oxidised manganese is tetravalent or trivalent. There are sources claim that Mn(OH)3 is the
brown precipitate, but hydrated MnO2 may also give the brown colour.

4 Mn(OH)2(s) + O2(aq) + 2 H2O → 4 Mn(OH)3(s)

The second part of Winkler test may reduces the acidities of the solution. The precipitate will
dissolve back into solution. Acid facilitates the coversion by the brown Manganese that contain
precipitate of the Iodide ion into elemental Iodine. The Mn(SO4)2 was formed by the acid converts
the iodide ions into iodine. Then, it is being reduced back to manganese (II) ions in an acidic medium.
Digit required x Digit multiplier = mg/L Dissolved Oxygen
Mn(SO4)2 + 2 I-(aq) → Mn2+(aq) + I2(aq) + 2 SO42-(aq)

Thiosulfate solution is used, with a starch indicator, to titrate the iodine.

2 S2O32-(aq) + I2 → S4O62-(aq) + 2 I-(aq)

Using the table below as reference :

Table 4.1 Reference table of standard water

Range (mg,L Sample Volume Titration Catalog Number Digit Multiplier

D.O.) (mL) Cartridge (N
Na2S2O3)
1-5 200 0.200 22675-01 0.01
2-10 100 0.200 22675-01 0.02
>10 200 2.000 14401-01 0.1

-8-
5.0 APPARATUS

Apparatus Chemical
Description Unit Cat.No

Alkaline Iodide-azide Powder

Pillows………………………................25/pkg…..............……..1072-68
Manganese Sulphate Powder
Pillows……..…......................25/pkg……..............................…..1071-68
300 mL BOD water
Sodium Thiosulfate Titration Cartridge. 0.2000 N....each….....22675-01
Starch Indicator Solution…...................................100ml MDB*...349-32
Sulfamic Acid Powder
Pillows………………………….............25/pkg…...…..............20762-68
Sample water

-9-
6.0 PROCEDURE

(USING A 300-mL BOD BOTTLE)

1. A water sample was collected in a clean 300-mL BOD bottle.

2. The contents of one Manganous Sulfate Powder Pillow and one Alkaline Iodide-Azide
Reagent Powder Pillow were added

3. The stopper was inserted immediately so air is not trapped in the bottle and it was inverted
several times to mix.

4. The floc in the solution was waited until settled and the bottle was inverted again for several
times and wait until the floc has settled.

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

6. A sample volume and Sodium Thiosulfate Titration Cartridge were selected corresponding to
the expected dissolved oxygen (D.O) concentration form.

7. A clean delivery tube was inserted into the titration cartridge. The cartridge was attached to
the titration body.

- 10 -
 8. The delivery knob was turned to eject a few drops of titrate. The counter was reset to zero
and the tip wiped.

9. A graduated cylinder was used to measure the sample volume. The sample was transferred
into a 250-mL Erlenmeyer flask.

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

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

12. The titration to a colourless end point was continued. The number of digits required was
recorded.

13. The total digits required multiply with Digit multiplier were calculated in mg/L dissolved
oxygen.

7.0 RESULTS

Dissolved Oxygen

Table 5: Result with Total Digit Required X Digit Multiplier = mg/L Dissolved oxygen

Range Sample volume Titration Catalog Digit (digit required)

(mg/L DO) (ml) Cartridge Number multiplier X

(N N2S2O3) (digit multiplier)

2-10 100 0.200 22675-01 0.02 6.86

- 11 -
8.0 CALCULATION

Experiment of oxygen using 300-mL BOD bottle

The range being use is between 2-10, by using 100mL sample volume and 0.02 digit multiplier. The
result obtain is:
mg/L Dissolved Oxygen = Digits required x Digit multiplier
= 343 mg/L X 0.02
= 6.86 mg/L Dissolved Oxygen

- 12 -
9.0 DISCUSSION

Dissolved oxygen (DO) is oxygen that is dissolved in water. There are many ways in dissolving oxygen
in water. For example, oxygen dissolves by diffusion from the surrounding air by aeration of water
that has tumbled over falls and rapids. An example of dissolving oxygen in water is given below.

Photosynthesis:

Carbon dioxide + Water → Oxygen + Carbon-rich foods

CO2 H2O O2 C6H12O6

Only green plants and some bacteria can dissolve oxygen through photosynthesis and similar
processes. Photosynthesis is daily process that happens in the water. In the morning, the rooted
plants and algae produced oxygen but at night, these plants use oxygen for their lives. This cause the
level of DO in water is low at night. Fish and aquatic animals cannot split oxygen from water (H2O) or
other oxygen-containing compounds.

- 13 -
 There were several steps that can be used in order to determine the concentration of
dissolved oxygen in water sample. Besides that, there also various types of chemical that can be used
to react with oxygen. Firstly, reagent pillow 1 which is Manganese Sulfate, MnSO4 was added into
sample water in BOD bottle. Through observation, we can see that the sample water form
flocculation after added MnSO4. MnSO4 was reacting with oxygen that present in water. Oxygen was
bounded with manganese and formed MnO2. This reaction formed a brownish solid that settles at
the bottom of the bottle. This process is called fixing the oxygen.

After that, other chemical that was added into the sample solution to react with sample
water is alkaline iodide-azide. There are three specific chemicals present in this reagent pillow which
are lithium hydroxide (LiOH), potassium iodide (KI) and sodium azide (NaN3). LiOH is one type of
basic. This means that water breaks them up to form the OH- ion and Li+ ion. Basically, function of
LiOH is as a catalyst to activate the binding process. Meanwhile, function of KI is as a dye. KI also
react with the added of sulfamic acid. The last reagent which is NaN3 is added to prevent conflict
reaction during the final titration. Sodium thiosulfate produces nitrite (NO2-) that conflicts with the
intended reaction. When the floc was formed, the floc was allowed to settle in the bottle to ensure
that the chemical were reacting completely.

At the addition of sulfamic acid (C6H1303NS), MnO2 that were produced was reduced to Mn2+.
MnO2 oxidized the iodine from the potassium iodide from I- to I2. This step causes the solution to
formed yellowish brown color. Here, the oxygen was fixed where all oxygen from water sample has
now been chemically modified to a form where it properties would not change when exposed to the
air.

MnO2 + 4H+ + 2I- = Mn2+ + I2 + 2H2O

As chemicals drops to enter the solution, the sodium separates from the thiosulfate ion.
Then, thiosulfate reacts with any iodine molecules which is available in the water. When the iodine
molecules react, it breaks up into I- ions until the color of solution changed to colorless.

2S2O32- + I2 = 2S4O62- + 2I-

The result showed that the mg/L of dissolved oxygen is 6.86 mg/L. The results value was in
range 2 to 10 mg/L. This mean it valid for digit multiplier times digit required to titrate the solution
into colorless. The result obtained is compared with standard quality of water that were proposed by

- 14 -
Jabatan Alam Sekitar Malaysia. It can be said that the type water sample was classified into classes II.
This mean that conventional treatment was required and recreational use with body contact.

Oxygen is the most important component to control the bioligical environment in water
sources. It is the end product of chemical and biological reaction in anaerobic condition. It give the
aesthetic effects such as undesirable colour, odor and taste. The reduction of oxygen in water can
cause a major effect on ecology and esthetic. Oxygen becomes the main cause of obvious pollution in
water that can be noticed visually. Oxygen appears in water from the photosynthesis process of
algae, rooted plants and from environment. Fishes and aquatic animals use oxygen everyday. The
minimum quantity of DO that needed in water sources is 2 mg/L. The level of DO that more than 4
mg/L is suitable for propagation for fish and aquatic wildlife.

The stability of oxygen in the water is dependent on some factors like prssure, temperature
and mineral concentration in te water. At pressure of 1 atmosphere with temperature of 20oC, the
DO level in water is 9.08 gram oxygen per 1m3 of water (Gary, 1999). The concentration of oxygen
decrease as the temperature increase. While, for concentration of salt, the concentration of the
oxygen will decrease when dissolved salt increases. The concentration of oxygen decrease when the
atmospheric pressure decreases.

- 15 -
10.0 CONCLUSION

The objectives of the experiment is to determined the concentration of dissolved oxygen in water
sample. From the calculation, the concentration value of dissolved oxygen was 6.86mg/L. This is
mean that the water sample is in class II when comply with standard quality water in Malaysia. This
standard quality of water also shows the quality of the water. The class II means that the water still
can be used for recreational activities. However, it still need conventional treatment.

Nowadays, the quality of water is very important because natural sources such as rivers and
lakes are used for many important purposed in our daily lifes. Life with low quality water can be
harm for health and life.

- 16 -
11.0 RECOMMENDATION

There are several recommendation that were proposed to improve the quality of the experiment.
First, allow the sample to overflow the bottle to ensure air bubbles are not trapped. Then, clean the
delivery tube with distilled water to ensure that there is no any other liquid that could mix up with
the liquid from the cartridge which can effects the result at the end of experiment. Other than that,
invert the BOD bottle properly to ensures the floc settle down properly.

- 17 -
12.0 REFERENCES

APHA et al., 1989. APHA, AWWA and WPCF, Standard Methods for the Examination of Water and
Wastewater. (17th edn ed.),, American Public Health Association, Washington, D.C.
(1989).

Bray et al., 1973. Bray J.T., Bricker O.P., and Troup B.N., Phosphate in interstitial waters of
anoxic sediments: Oxidation effects during sampling. Science, N.Y. 180 (1973), pp.
1362–1364

Carpenter S.R., Caraco N.F., Correll D.L., Howarth R.W., Sharpley A.N., and Smith V.H, Nonpoint
pollution of surface waters with phosphorus and nitrogen. Ecol Appl 83 (1998), pp. 559–568.

- 18 -
Chapman D. Water Quality Assessment. In: Chapman D on behalf of UNESCO, WHO and UNEP,
London: Chapman & Hall; 1992. 585p.

Förstner, 1990. Förstner U., Inorganic sediment chemistry and elemental speciation. In: Baudo R,
Giesy J.P., and Muntau H., Editors, Sediments: Chemistry and Toxicity of In-Place Pollutants, Lewis,
Chelsea, Mich (1990), pp. 61–105.

13.0 APPENDICES

- 19 -
- 20 -
 2 3

Flow diagaram of dissolved oxygen experiment

- 21 -