1.

ABSTRACT
Biochemical oxygen demand, or BOD, measures the amount of oxygen consumed by
microorganisms in decomposing organic matter in stream water. BOD also measures the
chemical oxidation of inorganic matter for example the extraction of oxygen from water via
chemical reaction. The rate of oxygen consumption in a stream is affected by a number of
variables: temperature, pH, the presence of certain inds of microorganisms, and the type of
organic and inorganic material in the water.
!"ide #odification of $inler #ethod is one simple way in determining amount of Dissolve
Oxygen %DO& in water. This multi step chemical method involves the adding of chemicals such
as #anganous 'ulfate (owder (illow and !laline )odide*!"ide +eagent (owder (illow
followed by 'ulfamic (owder (illow , that react with water and ,fixes- it. .ertain reagents will
be placed in sample water that is taen from a water supply. !ny observation during these
periods shows the presence or absence of DO in water. The adding of these reagents will produce
a prepared sample which will be titrated with sodium thiosulfate using a titrator body with
titration cartridge until a pale yellow solution is formed. ! simple calculation later on will
determine the exact amount of DO in the sample water.
1
2.INTRODUCTION
The stream system both produces and consumes oxygen. )t gains oxygen from the atmosphere
and from plants as a result of photosynthesis. +unning water, because of its churning, dissolves
more oxygen than still water, such as that in a reservoir behind a dam. +espiration by a/uatic
animals, decomposition, and various chemical reactions consume oxygen.
$astewater from sewage treatment plants often contains organic materials that are decomposed
by microorganisms, which use oxygen in the process. %The amount of oxygen consumed by these
organisms in breaing down the waste is nown as the biochemical oxygen demand or BOD.
Other sources of oxygen*consuming waste include stormwater runoff from farmland or urban
streets, feedlots, and failing septic systems.
Oxygen is measured in its dissolved form as dissolved oxygen %DO&. )f more oxygen is
consumed than is produced, dissolved oxygen levels decline and some sensitive animals may
move away, weaen, or die.
DO levels fluctuate seasonally and over a 01*hour period. They vary with water temperature and
altitude. .old water holds more oxygen than warm water and water holds less oxygen at higher
altitudes. Thermal discharges, such as water used to cool machinery in a manufacturing plant or
a power plant, raise the temperature of water and lower its oxygen content. !/uatic animals are
most vulnerable to lowered DO levels in the early morning on hot summer days when stream
flows are low, water temperatures are high, and a/uatic plants have not been producing oxygen
since sunset.
Total dissolved gas concentrations in water should not exceed 223 percent. .oncentrations above
this level can be harmful to a/uatic life. 4ish in waters containing excessive dissolved gases may
suffer from 5gas bubble disease56 however, this is a very rare occurrence. The bubbles or emboli
bloc the flow of blood through blood vessels causing death. 7xternal bubbles %emphysema& can
also occur and be seen on fins, on sin and on other tissue. !/uatic invertebrates are also
affected by gas bubble disease but at levels higher than those lethal to fish.
!de/uate dissolved oxygen is necessary for good water /uality. Oxygen is a necessary element
2
to all forms of life. 8atural stream purification processes re/uire ade/uate oxygen levels in order
to provide for aerobic life forms. !s dissolved oxygen levels in water drop below 9.3 mg:l,
a/uatic life is put under stress. The lower the concentration, the greater the stress. Oxygen levels
that remain below 2*0 mg:l for a few hours can result in large fish ills.
The concentration of dissolved oxygen in water is affected by many factors including ambient
temperature, atmospheric pressure, and ion activity. !ccurate data on the concentration of
dissolved oxygen %DO& in environmental water resources are essential for documenting changes
that result from natural phenomena and human activities. #any chemical and biological
reactions in ground water and surface water depend directly or indirectly on the amount of
available oxygen. Dissolved oxygen is necessary in a/uatic systems for the survival and growth
of many a/uatic organisms and is used as an indicator of the health of surface*water bodies.
3.OBJECTIVE
The main purpose for doing this experiment are :
To determine the dissolved oxygen in water sample %lae water&
To ascertain whether the /uality complies with the #alaysian $ater 'tandards
3
1. THEORY
There are various type of element contained in a water. One of the elements is oxygen. The
stream system both produces and consumes oxygen. Oxygen dissolves in the stream by
diffusion of oxygen from the atmosphere and also by photosynthesis from plants. Running water
will dissolve much more oxygen than a still water such as reservoir. Oxygen is measured in its
dissolved form as dissolved oxygen (O!. O levels differ seasonally and over a 2"#hour
period. They vary with water temperature and altitude. $old water contains more oxygen than
warm water and water contained less oxygen at higher altitudes.
O is measured either in milligrams per liter (mg%&! or 'percent saturation.' (illigrams per
liter is the amount of oxygen in a liter of water. )ercent saturation is the amount of oxygen in a
liter of water relative to the total amount of oxygen that the water can hold at that temperature.
O samples are collected using a special *O bottle. issolved oxygen can be measured
either by using the +in,ler method or by using a meter and probe.
The +in,ler method involves filling a sample bottle completely with water. The dissolved
oxygen is then added by a series of reagents that form an acid compound that is titrated.
Titration involves the drop by drop addition of a reagent that neutrali-es the acid compound and
causes a change in the colour of the solution. The point at which the colour changes is the
endpoint of the titration and is e.uivalent to the amount of oxygen dissolved in the sample.
'odium thiosulphate is the reducing agent normally used and starch solution is used to
determine the end point of the titration. !ll reactions in the determination of oxygen involves
oxidation and reduction. However, starch is used as end point indicator, and forms a starch*
iodine complex with iodine from dilute solutions to produce a blue colour and change to a colour
less form when the iodine is all reduced to iodide ion. The reaction involved in $inler
procedure is given as follow.
#n
0;
; 0OH
*
#n%OH&
0
#n
0;
; 0OH
*
; < O
0
#nO
0
; H
0
O
#n%OH&
0
; < O
0
#nO
0
; H
0
O
"

The oxidation of #n%))& to #nO

0
, sometime called fixation of the oxygen, occurs slowly,
particularly at low temperatures. 4urthermore, it is necessary to move the flocculated material
throughout the solution to enable all the oxygen to react. =nder low pH conditions the #nO
0
oxidi"es )
*
to produce )
0
#nO
0
; 0)
*
; 1 H
;
#n
0;
; )0 ; 0 H
0
O
)odine is the rather insoluble in water, but forms a complex with the excess iodide present to
reversibly form the more soluble tri*iodate, thus preventing escape of )0 from solution
)
0
; )
*
)
>*
The sample is then titrated against a standard thiosulphate solution, using starch as indicator
towards the end. 4rom the titer value, results can be interpreted directly in terms of milligrams
per liter of DO.
?23@
5.METHODOLOGY
APPARATUS
>33mA BOD bottle
Blove
/
Boogles
Dissolved oxygen field it
'odium Thiosulfate Titration .artridge
Delivery tube
Braduated cylinder
093 mA 7rlenmeyer flas
(ipette
DO #eter calibration
MATERIALS
#anganous 'ulfate (owder (illow
!laline )odide*!"ide +eagent (owder (illow
'ulfamic !cid (owder (illow
'odium Thiosulfate
'tarch indicator solution
6.PROCEDURE
2. ! water sample in a clean >33 mA BOD bottle was collected. The sample was allowed to
0
overflow the bottle for 0*> minutes to ensure the air bubbles were not trapped.
0. 8ext the sample was added with one #anganous 'ulfate (owder (illow and one !laline
)odide*!"ide +eagent (owder (illow.
>. ! stopper was inserted immediately without trapping air inside the bottle. The bottle was
inverted several times to mix the solution.
1. !fter the bottle was inverted several times,waited until the floc settles and a clear
solution were obtained.
9. The stopper was removed and the content were added with one 'ulfamic (owder (illow.
The stopper was replaced without trapping the air inside the bottle and then again the
prepared sample was inverted several times.
C. ! sample volume and 'odium Thiosulfate Titration .artridge were selected
corresponding with expected dissolved oxygen %DO& concentration.
D. Then a clean delivery tube was inserted into the titration cartridge to the titrator body.
!fter that, the delivery nob was turned to inEect a few drops of titrant. The counter is
resetted to "ero and the tip was wiped.
F. ! graduated cylinder was used to measure the sample volume and the sample was
transferred to 09mA 7rienmeyer flas.
G. The delivery tube was placed into the solution and the flas is swirled while the solution
was titrated with sodium thiosulfate to a pale yellow colour.
23. 8ext, two 2 mA dropper of 'tarch )ndicator solution were added and the mixture was
swirled.
22. The titration was continued to get a colourless end point. The number of digits re/uired
were recorded.
20. 4inally, dissolved oxygen were calculated and being tabulated in a table.
1
7.RESULTS
Holume
'ample
water
% Aae
water &
Observation Holume
'odium
Thiosulphate
used %mA&
Dissolve
oxygen
%DO& levels
%mg:A&
#anganous
sulphate ;
iodide a"ide
'ulfamic acid starch
>33mA
Brown
orange
floc
I4loc dissolve
I4orm yellow
colour solution
'olutions
turns blue
02D 1.>1
C3mA Brown
orange
floc
I4loc dissolve
I4orm orange*
yellow colour
solution
'olutions
turns blue
10 1.20
Solution Observation/conclusion
!J$ater sample ; 2 #anganous (roduce orange*brown flocculent. )ndicates that oxygen
2
'ulfate !laline )odide ; 2 !"ide
+eagent (owder (illow.
is present in sample water.
BJ! ; 2 'ulfamic !cid (owder
(illow
4locculent dissolved and leaves yellow coloured
solution. )ndicates that oxygen is present in sample
water.
.J233mA of B ; 'odium
Thiosulfate %titration&
Drop by drop until solution turns pale yellow.
DJ. ; two 2mA droppers of
'tarch )ndicator 'olution
'olution turns dar blue. .ontinued titration until
solution turns colourless. 8umber of digits re/uired is
recorded.
8.CALCULATIONS
8umber of digits re/uired x digit multiplier J mg:A dissolved oxygen
3
Holume of water sample
% lae water &
Holume of 'odium
Thiosulphate used %mA&
Dissolve Oxygen %DO&
level %mg:A&

>33 mA 02D J 02D 3.30
J 1.>1
C3 mA 10 J 10 3.2
J 1.0
DISCUSSION
The main obEective of this experiment is to determine the dissolved oxygen in water sample
lae water. The dissolved oxygen in water sample is determined by using the win,ler a-ide
method. *ased on an article on 4ow To (easure issolved Oxygen by epartment of 5cology6
7tate of +ashington and an article produced by an online botanical encyclopaedia6 it shows that
18

win,ler method is the most precise and reliable titrimetric procedure for dissolved oxygen
analysis.
The win,ler method is a techni.ue which uses titration to determine the dissolved
oxygen in freshwater systems. 9ccording to an article by (onica :. *ru,ner in (icrobial &ife
5ducational Resource (2811!6 dissolved oxygen is used as an indicator of the health of a water
body6 where higher dissolved oxygen concentrations are correlated with high productivity and
little pollution. The dissolved oxygen analysis can be used to determine the health or cleanliness
of a la,e or stream6 the amount and type of biomass a freshwater systems can support6 and the
amount of decomposition occurring in the la,e or stream.This experiment was conducted to
verify the amount of dissolve oxygen contain in various sample water using !"ide #odification
of $inler #ethod.
The !"ide used in this experiment inhibits the interference of nitrogen ion during the redox
reaction by oxygen and iodine.
The addition of #anganous 'ulfate amd !laline )odide +eagent (owder (illow produce an
orange*brown precipitate due to the reduction of oxygen by #nK.
0 #n
0;
; 1 OH
*
; O
0
0#nO
0
%s& ; 0 H
0
O %brown precipitate&

'ulfamic !cid (owder (illow, H8'O was added and cause #nO to oxidise ).
Therefore, the precipitate dissolve and leave an orange*brown precipitate. The orange*brown
precipitate indicates the presence of oxygen in the sample water.
#nO
0
%s& ; 1 H
;
; 0 )
*
)
0
; #n
0;
; 0 H
0
O %orange*brown solution&

! definite amount of this sample is measured and selected to be titrated with 'odium
thiosulphate solution. Before titrating, a few drops were eEected from the tube to ensure there are
no bubbles traped that may cause an error. The titration tube is immersed directly into the
solution to ensure full reaction. The container of the solution is swirl throughout the titration to
speed up the rate of reaction.
11
The ) which present as triiodide ion, ) is titrated with 'odium Thiosuphate until it turns
pale yellow.
1 8a
0
'
0
O
>
; 0)
0
08a
0
'
1
O
C
; 18a) %pale yellow&

'tarch indicator is added and solution turns dar blue because of the presence of iodine in the
solution. !s nown, the #nO formed earlier s directly proportional to the amount of oxygen
present in the sample water. 8ow, the amount of iodine present is directly proportional to the
#nO present. Therefore, by titrating iodide with sodium thiosulphate until the solution turns
colourless will indicates the amount of DO. The amount of DO was calculated using this value,
which are 1.>1 mg:A.
;or the first step in this experiment6 all water samples were collected in the +ater
7ampling *ottle also ,nown as *O bottle and stoppered brim full with sample water. The
sample water was allowed to overflow a little before capping as this is crucial to avoid aeration
and additional oxygen into the water samples. Then6 (anganous 7ulphate )owder )illow and
<odide 9-ide )owder )illow was added to the water samples and inverted which resulting to
formation of brown orange solution with precipitate called floc indicating the presence of
dissolved oxygen. 9ccording to issolved Oxygen +ater =uality Test >it <nstruction (anual by
&a (otte 6 immediately upon formation of the precipitate6 the oxygen in the water oxidi-es an
e.uivalent amount of the manganous hydroxide to brown#coloured manganic hydroxide. ;or
every molecule of oxygen in the water6 four molecules of manganous hydroxide is converted to
manganic hydroxide.
Then a 7ulfamic 9cid )owder )illow was added to the water samples and the bottle was
inverted several times. This reagent is added to dissolve the floc and leaving only the orange#
yellow solution in the bottle. The acid converts the manganic hydroxide to manganic sulfate. 9t
this point6 the sample is fixed and can be stored for up to eight hours if ,ept in a cool6 dry place.
7imultaneously6 iodine from <odide 9-ide 7olution is oxidi-ed by manganic sulfate6 releasing
free iodine into the water. 7ince the manganic sulfate for this reaction comes from the reaction
between the manganous hydroxide and oxygen6 the amount of iodine released is directly
12
proportional to the amount of oxygen present in the original sample. The release of free iodine is
indicated by the sample turning a orange#yellow colour. 9fter that6 188 ml of this water sample
solution was transferred into a conical flas, and titrated with sodium thiosulphate solution and
swirled until the yellow colour turns pale. Then6 starch was added as an indicator and the
solution turns blue blac, upon addition. The water sample in the conical flas, was further titrated
until the colour turns colourless which indicates the endpoint of the titration. The sodium
thiosulfate reacts with the free iodine to produce sodium iodide. +hen all the iodine has been
converted the sample changes from yellow to colourless. The reason for titrating until the
solution turns pale earlier is to let the starch stays dar, blue right up until it goes clear6 unli,e
most titrations where the colour gradually moves toward the endpoint. Therefore6 it is easy to
become complacent during the titration and add an excess amount of titrant which overshooting
the endpoint6 thin,ing that you are far from the endpoint because the colour is not changing.
9nother reason was starch can be partially decomposed by a large amount of iodine. Therefore6
the starch should not be added until the bul, of the iodine has been reduced.
The amount of dissolved oxygen in la,e water is calculated to be ".3" mg%& when using
388 m& of water sample 6 while ".2 mg%& when using 08 m& water sample 6 whereas de#ioni-ed
water contained the most dissolved oxygen which is ".3" mg%&. *ased on the )roposed
?ational +ater =uality 7tandards for (alaysia prepared by @ayasan 7abah ;orest
(anagement 9rea6 these water samples can be categori-ed into a few classes which are class
<6 <<96 <<*6 <<<6 <A6 and A based on their issolved Oxygen (O! value (see appendix!.
;or type < classes the value of O ranged from 1 and above. *ased on this experiment6
it was proven that one of the water sample6 which is de#ioni-ed water falls into this category.
This type < class represents the excellent water .uality which meets the most stringent
re.uirement for human health and a.uatic life protection.
?evertheless6 our experiment is nowhere near perfect. There are some errors made during
the experiment. <n my opinion6 the dissolved oxygen value of la,e water is not an actual value
due to the presence of air bubbles inside the water sample in the water sampling bottle. *esides
that6 the water sample is not collected at near the middle of the la,e or at armBs length below the
water surface. This is because dissolved oxygen concentrations may change drastically in la,es
depending upon depth and distance from shore. The starch solution added is also not
accurately of 1ml for each drop and there might also be the presence of air bubbles in the
delivery tube6 resulting in the accuracy of the results obtained. These problems adds to reasons
13
of my perspective that the results obtained were not of the precise value.
10.CONCLUSION
=sing !"ide #odification of $inler #ethod, the DO of various sample water is nown.
!ddition of !"ide will inhibit the interference of nitrogen ion during the redox reaction of
reagents in the water sample. .ompared to the 8ational $ater Luality 'tandards for #alaysian,
the sample water has the rate of DO among the ))) class.
The means that contribute as class ))) is:
Defined with the primary obEective of protecting common and moderately tolerant a/uatic
species of economic value. $ater under this classification may be used for water supply with
extensive : advance treatment. This class of water is also defined to suit livestoc drining needs.
The dissolved oxygen contain in lae water is determined to be C.1 mg:A, tap water C.0mg:A,
drain water F.C mg:A, deioni"ed water D.1mg:A. The lae water and tap water is determined to
be polluted.
11.RECOMMENDATIONS
Here are some of the recommendations regarding to this study, before conducting experiment
students should put on gloves and goggles. This is to avoid contamination. !lways hold the
dropper perpendicular to and approximately one inch above the sample bottle so that drop si"e
will be consistent. 8ext, avoid the issue of bubble formation by using the dissolution medium
after e/uilibration at >D. for 01 hour in a water bath. Aogic behind this approach is when
medium is e/uilibrated for sufficiently long time at >D. and when transfer to dissolution bath, no
temperature changes, no change in dissolved gasses solubility, thus no bubble formation. The
titrator plunger has a tendancy to stic at the bottom or wherever it has been stored. To help
avoid this, always store the titrator with plunger not fully depressed. Before use, push on the
plunger to loosen it before pulling up. Besides, it is suggested to use digital titrator to get
1"
accurate and precise result. 7xperiment can be /uite variable so collect sufficient field of water
and split replicates %23 to 03 percent& to provide an estimate of method variability
12.REFFERENCES
2 .Brown, Theodore A., H. 7ugene Ae#ay, Mr., and Bruce 7. Burston. %033C&.hemistry: The
.entral 'cience. 23th ed. =pper 'addle +iver, 8M: (earson 7ducation, )nc.,
0. Determination of dissolved oxygen by $inler titration.
httpC%%www.core.org.cn%?R%rdonlyres%5arth##9tmospheric##and#)lanetary#7ciences%12#
831Danuary##<9)##2880%3/;9;;$3#13/*#"200#33*"#21"";1"2188"%8%dissolvedEoxygen.pdf
search on : 23 may 032>
>. Aide, D. +. %7d.& %2GG3&. .+. Handboo of .hemistry and (hysics %D3th 7dn.&. Boca +aton
%4A&:.+. (ress.
1. 'awyer, .. 8., #c.arty, (. A., and (arin, B. 4.%033>& .hemistry for 7nvironmental
7ngineering, 9th ed., #cBraw Hill
9. 9nonymous (2811!6 4ow to (easure issolve Oxygen6 epartment of 5cology 7tate of
+ashington retrieved from http:::www.ecy.wa.gov
search on C 18 may 2813
0. 'tudy and )nterpretation of the .hemical .haracteristics of 8atural $ater,%2GD3& =nited
'tates Beological 'urvey, $ater 'upply (aper 21D>,
1/
13.APPENDICES
10
11