1

TABLE OF CONTENTS
PAGE

1.1

ABSTRACT/SUMMARY

1.2

INTRODUCTION

1.3

AIMS

1.4

THEORY

1.5

METHODOLOGY/PROCEDURE

1.6

APPARATUS AND MATERIALS

1.7

RESULTS

1.8

CALCULATIONS

10

1.9

DISCUSSION

10

2.0

CONCLUSIONS

12

2.1

RECOMMENDATIONS

12

2.2

REFERENCE

13

2.3

APPENDIX

13

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ABSTRACT
From this experiment , we have determined the amount of dissolved oxygen from the lake water
in Shah Alam sample that we had collected. Through the observations, we noticed that the lake
water sample that we had collected contain the oxygen, dissolved oxygen by it properties
showing the orange-brown color with precipitates when added with Manganese Sulphate Powder
Pillow with Alkali Iodide Azide Reagent Powder Pillow. When added with sulfonic acid powder,
the water sample have a color yellow, which both of the results proved the existence of oxygen in
the water sample. Using a few series of calculations, we have determined the amount of
dissolved oxygen in the lake water sample.

INTRODUCTION
Oxygen saturation or dissolved oxygen (DO) is a relative measure of the amount of oxygen
dissolved in a medium. DO is naturally present in water and tends to be manipulated in order to
suit the specific applications. For an example, DO level in drinking water improves the taste.
Other than that, high DO level is important for every living organism. Aquatic organisms needed
appreciable level of DO to exist in water. However, the industry tends to minimize the DO level
to reduce the maintenance costs and in order to reserve it pipelines from corrodes.
The level of DO in water depends on many physical, chemical, and biochemical factors
which is aeration, wing, velocity of water flow, algae, temperature, atmospheric pressure, organic
compounds, bacteria, salt content and animals. Oxygen is produced

by algae during

photosynthesis under the presence of sunlight. Nevertheless, this process is really not an efficient
means of oxygenating water because some of the oxygen formed by photosynthesis during the
daylight hours is lost at night when the algae consume oxygen as a part of their metabolic
processes. When the algae die, the degradation of the biomass also consumes oxygen.
Besides, temperature also gives a significant impact on the solubility of oxygen in water.
Increasing the temperature will give arose to the DO concentration in water. It is important to
distinguish between oxygen solubility, which is the maximum DO concentration at the

3
equilibrium, and the actual concentration of the DO level. The actual concentration of the DO
level which is generally not the equilibrium concentration and is limited by the rate at which the
minimal temperature where the oxygen dissolve. Water will become saturated with oxygen at
25oC, the minimal temperature.
So in this experiment we will determine the amount of the dissolved oxygen in water by
using the selected procedure that is Azide Winkler Method. Using this method, we will checked
the dissolve oxygen in the sample and it must be comply with Malaysian Standard of Water
Quality.

AIMS
To proposed method to check dissolve oxygen in water sample. To determine the number and
types of organisms living in that body of water by calculate amount of dissolved oxygen in water
sample.

THEORY
Winkler test is used in this experiment in order to measure the concentration of dissolved oxygen
(DO) in a lake water sample. In this method, excess manganese (II) sulphate will react with the
alkali iodide-azide reagent that is an iodide ion (I-) and potassium hydroxide ion (OH-) in water
to form a white precipitate Mn(OH)2. Alkali iodide azide also composed of NaN3 that is an alkali
azide compound that will eliminated the nitrile interference that can interfere the reaction of
reducing or oxidizing substances in this test.
MnSO4 + 2KOH

Mn(OH)2 + K2SO4

(1)

If oxygen is present in the water, the Mn(OH)2 will react further to form an orangebrown precipitate, manganic oxide (MnO(OH)2) but if the oxygen is not present in the water
sample, the color of the white precipitate will remain unchanged and no further reaction will
occurs. (Ruth F. Weiner, 2003)

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2Mn(OH)2 + O2

2MnO(OH) .. (2)

Sulfamic acid powder pillow, H3NSO3 is added, which dissolves the manganic oxide and in
together with the potassium iodide, KI added earlier, forms iodine (I2), which gives color to the
water sample.
2Mn(OH)2 + 4H3NSO3
2Mn(SO4)2 + 4KI

2Mn(SO4)2 + 6H2O (3)

2MnSO4 + 2K2SO4 + 2I2- .(4)

Then by referring to the table below, a sample of volume of the solution is taken and poured into
a graduated cylinder.
Range(mg/L

Sample

Titration

D.O)

volume(mL)

cartridge

200
100
200

(NNa2S2O3)
0.2000
0.2000
2.000

1-5
2-10
>10

Catalog number

Digit multiplier

22675-01
22675-01
14401-01

0.01
0.02
0.1

The quantity of Iodine in the water sample is measured by the method of titrating with
sodium thiosulphate (Na2S2O3) until the orange color of the Iodine will turns to pale yellow as
pale as the color of yellow plastic straw.
4Na2S2O3 + 2I2

2Na2S4O6 + 4NaI . (5)

At the end of the titration, starch is added to measure if there is any unreacted iodine, I 2 in
the solution. The starch will gives a dark blue color in the presence of the I 2 and it will becomes
more obvious at the end of the experiment and it will indicates the endpoint of the experiment.
Then it will be titrated again with sodium thiosulphate solution until the color of the solution
becomes colorless.
The quantity of MnO(OH)2 which is still available formed in the first step of the reaction
is directly proportional to the dissolved oxygen, and the amount of I 2 formed in the second

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equation is directly proportional to the MnO(OH)4. Thus in can be conclude that the titration of
the sodium thiosulphate solution measures a quantity of iodine directly related to the original
dissolved oxygen concentration. Therefore, we can calculate the amount of sodium thiosulphate
used by recording the digits at the titration and multiply it by the digit multiplier given at a
sample volume we had used like in table shown before.
In the 2nd test, we used oxygen powder pillow 1 and 2, in order to trap the oxygen that
gives out the yellow color of oxygen and then oxygen powder pillow 3 will be inserted to
dissolve any precipitate and titrate it with sodium thiosulphate as in test 1.
Dissolved oxygen(mg/L) = digits required 0.02
Using this formula, we can determine the dissolved oxygen in the water sample.

PROCEDURE
Azide Modification of Winkler Method (using 300mL BOD bottle)
1. A water sample is collected in a clean 300ml BOD bottle.
2. One Manganous Sulfate Powder Pillow and one Alkaline Iodide-Azide Reagent Powder
Pillow are added into the BOD bottle at the same time.
3. Then, immediately insert the stopper so that the air is not trapped in the bottle. To mix
well, invert the bottle several times.
4. Wait until the floc in the solution has settled. Again invert the bottle several times and
wait until the floc has settled.
5. The stopper is removed and one Sulfamic Acid Powder Pillow is added in the bottle.
6. The stopper is replaced without trapping air in the bottle and invert the prepared sample
several times to mix.
7. A sample volume and Sodium Thiosulphate Titration Cartridge is selected corresponding
to the expected dissolved oxygen (DO) concentration from Table 1.
8. The delivery tube is cleaned properly before inserted into the titration cartridge. Attached
the cartridge to the titrator body.
9. The delivery knob is turned to eject a few drops of titrant.
10. Reset the counter zero and wipe the tip.

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11. Measure 200ml of sample volume by using graduated cylinder. Transfer the sample into a
250 ml Erlenmeyer flask.
12. The delivery tube tip is placed into the solution and swirl the flask while titrating with
Sodium Thiosulfate to a pale yellow colour.
13. Two 1ml droppers Starch Indicator Solution is added. Swirl to mix well.
14. The titration is continued until to a colourless end point.
15. The numbers of digits required are recorded.
16. Dissolved oxygen are calculated by using this formula:
Dissolved oxygen(mg/L) = digits required digit multiplier

TABLE 1
Range(mg/L

Sample

Titration

Catalog

D.O)

volume(mL)

cartridge(NNa2S2O3

number

1-5
2-10
>10

200
100
200

)
0.2000
0.2000
2.000

22675-01
22675-01
14401-01

Digit multiplier

0.01
0.02
0.1

Using a 60ml BOD bottle

1. A sample water is collected in a clean 60ml BOD bottle.
2. One Dissolved Oxygen 1 Reagent Powder Pillow and one Dissolved Oxygen 2 Reagent
Powder are added into the BOD bottle at the same time.
3. Then, immediately insert the stopper so that the air is not trapped in the bottle. To mix
well, invert the bottle several times.
4. Wait until the floc in the solution has settled. Again invert the bottle several times and
wait until the floc has settled.
5. The stopper is removed and one Dissolved Oxygen 3 Reagent Powder Pillow is added in
the bottle.
6. Replace the stopper without trapping air in the bottle and invert the prepared sample
several times to mix.
7. 20 ml of prepared sample is measured and transferred it to a 250ml Erlenmeyer flask.
8. Attach a clean delivery tube to a 0.2000 N Sodium Thiosulfate Titration Cartridge. Twist
the cartridge onto the titrator body.
9. Eject a few drops of titrant by turning the delivery knob. Reset the counter to zero and
wipe the tip.
10. The prepared solution is titrated with 0.2000 N Sodium Thiosulfate until the sample
changes from yellow to colourless.

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11. The number of digits are recorded.
12. The Dissolved Oxygen is calculated by using this formula:
Dissolved Oxygen(mg/L) = Digits required 0.1
APPARATUS/MATERIALS
Apparatus:
60ml BOD bottle
300ml BOD bottle
Measuring cylinder
250ml Erlenmeyer flask
Straight-stem delivery tube
Beaker
0.2000 N Sodium Thiosulfate Titration Cartridge
Dropper
Stopper

Reagent :

Sample water
Dissolved Oxygen1 Reagent Powder Pillow
Dissolved Oxygen 2 Reagent Powder Pillow
Dissolved Oxygen 3 Reagent Powder Pillow
0.2000 N Sodium Thiosulfate
ManganousSulfate Powder Pillow
Alkaline Iodide-Azide Reagent Powder Pillow
Sulfamic acid Powder Pillow
Two 1mL droppers of starch indicator solution.

RESULT
To determine the amount of dissolve oxygen in the water sample.
(300ml BOD bottle)
Reagent
Manganous sulphate powder

Observation
Flocculent precipitate form,

pillow + Alkaline Iodide

colorless water turn into

Azide reagent powder pillow.

orange-brown color

Sulfonic Acid Powder

orange-brown colour turn into

Conclusion
Oxygen present

Oxygen present

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clear yellow colour
Digits required = 203
Digits multiplier = 0.01
Dissolved oxygen (mg/L) 0.0 = digit required x digit multiplier
= 203 x 0.01
= 2.03 mg/L
Range = 1-5 mg/L
( 60ml BOD bottle )
Reagent
One dissolve oxygen 1

Observation
Flocculent precipitate form,

reagent powder pillow + one

colourless water turn into

dissolve oxygen 2 reagent

orange-brown colour

Conclusion
Oxygen present

powder pillow.
One dissolve oxygen 3

orange-brown colour turn into

powder pillow

clear yellow colour

Digits required = 37
Digits multiplier = 0.1

Dissolved oxygen (mg/L) 0.0 = digit required x digit multiplier

= 37 x 0.1
= 3.7 mg/L dissolve oxygen

Oxygen present

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Range = 2-10 mg/

CALCULATIONS
For 300 mL BOD Bottle
Dissolved oxygen (mg/L) 0.0 = digit required x digit multiplier
= 203 x 0.01
= 2.03 mg/L
For 60 mL BOD Bottle
Dissolved oxygen (mg/L) 0.0 = digit required x digit multiplier
= 37 x 0.1
= 3.7 mg/L dissolve oxygen

DISCUSSION
Basic water properties 1 experiment consists of two test that must be conducted to determine the
dissolved oxygen in water. The method that used in this experiment is the Azide Modification of
Wrinkler Method. Two test that must be conducted in this experiment are testing the water
sample which is lake water by using a 300-mL BOD bottle and also using a 60-mL BOD bottle.
The type and location of the water sample collected were same but they had different in the
volume of water sample used.

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We conducted the first test of the lake water by using the 300-mL BOD bottle. The
sample water must be collected into a 300-mL BOD bottle with no air bubble trapped in it. Then,
one Manganous Sulfate Powder Pillow and one Alkaline iodideAzide Reagent Powder Pillow
were added into the bottle. The stopper of the bottle was immediately inserted to prevent the
bubble air trapped and it was inverted several time. As the result, the color of the sample water
changed from colorless to orange brown color with flocculent precipitate. These result can be
seen when the oxygen was present in the water but if not, the water sample would turned to
white .The solution in the bottle was added by one Sulfamic Acid Powder Pillow and the stopper
was inserted immediately. The bottle once again had been inverted several times to mix the
mixture. From that, the floc dissolved and leaved the yellow color if the oxygen present. 100-mL
of the solution has been measured and taken into an Erlenmeyer flask. Then, the solution is
titrated with Sodium Thiosulfate Titration Cartridge to find the expected dissolved oxygen (D.O).
1-mL of Starch Indicator Solution is added to the solution and swirled. In the same time, titration
continued until the dark blue color (effect from the starch) turned to colorless. The quantity of
the Sodium Thiosulphate used is calculated to indicate the dissolved oxygen in water .The digits
required (203) must be multiplied by digit multiplier (0.01) to find the dissolved oxygen which
was 2.03 mg/L and it follow the range that stated in TABLE 1.
The second test was conducted by using 60-mL BOD bottle. The water sample was
collected into the bottle and added by one Dissolved Oxygen Reagent Powder Pillow and one
Dissolved Oxygen 2 Reagent Powder Pillow. After that, the bottle was inverted for several time
then the color of solution turned from colorless to orange-brown which shown that oxygen
present. Then, one Dissolved Oxygen 3 Powder Pillow. The bottle was inverted several time
again until the floc dissolved and leaved the yellow solution. 100-mL of the solution was taken
from it and put into the 250-mL Erlenmeyer Flask. The solution then titrated by 0.2000 N
Sodium Thiosulfate Cartridge until the solution turned from yellow to colorless. The quantity of
the Sodium Thiosulfate used is calculated and recorded. To measure the dissolved oxygen in the
water sample, the digital required of Sodium Thiosulfate which is 37 was multiplied by 0.1 .So,
the dissolved oxygen calculated here was 3.7 mg/L.
In this experiment, the experimenter must be heedful while inserting the water sample
into the bottle at the beginning. It must be done successfully to prevent the air bubble trapped in

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the bottle. If there was air bubble, it would disturb the result. Other than that, when 0.2000 N
Sodium Thiosulfate was added, the solution in Erlenmeyer flask must be swirled. So that, the
colour would change completely when the volume of Sodium Thiosulfate is enough.

CONCLUSION
For experiment using 300mL BOD bottle by using the reagent between Manganous sulphate
powder pillow with Alkaline Iodide Azide reagent powder pillow, flocculent precipitate and
colourless water turn into orange-brown colour are formed. Therefore, the oxygen was presented.
Meanwhile for reagent Sulfonic Acid Powder, orange-brown colour turn into clear yellow colour
formed. These also mean that oxygen was presented. The oxygen dissolve is 2.03 mg/L. The
result shows that the water sample content complies with Malaysian Standard water which the
value of oxygen dissolves still in range 1 until 5 mg/L.
For experiment using 60mL BOD bottle by using the reagent between one dissolve oxygen
1 reagent powder pillow with one dissolve oxygen 2 reagent powder pillow, flocculent
precipitate and colourless water turn into orange-brown colour are formed. Therefore, the oxygen
was presented. Meanwhile for reagent One dissolve oxygen 3 powder pillow, orange-brown
colour turn into clear yellow colour formed. These also mean that oxygen was presented. The
oxygen dissolve is 3.7 mg/L. The result shows that the water sample content complies with
Malaysian Standard water which the value of oxygen dissolves still in the range 2 until 10 mg/L.

RECOMMENDATION
1. The experiment must be heedful while inserting the water sample into the bottle at the
beginning to prevent air bubble trapped in the bottle
2. We must waiting until the flock in the solution to make sure the reaction of the sample
and reagent are completed.
3. We must swirl the Erlenmeyer flask to make sure the reaction is mix completely after
adding Sodium Thiosulphate.

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REFERENCE
1.
2.
3.
4.

http://wiki.answers.com/Q/Explain_the_difference_between_cohesion_and_adhesion
http://www.watereducation.utah.gov/waterscience/Properties/default.asp
http://www.scribd.com/doc/39166232/Basic-Water-Properties-2-Experiment
http://serc.carleton.edu/microbelife/research_methods/environ_sampling/oxygen.html

APPENDIX
TABLE 1
Range(mg/L

Sample

Titration

Catalog

D.O)

volume(mL)

cartridge(NNa2S2O3

number

1-5
2-10
>10

200
100
200

)
0.2000
0.2000
2.000

22675-01
22675-01
14401-01

Digit multiplier

0.01
0.02
0.1

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