UNIVERSITY OF

NAIROBI
PUBLIC HEALTH ENGINEERING
FCE 581 LABS
NJOROGE KAGWI MAURICE
F16/1334/2010

FCE 581

F16/1334/2010

2014/2015

EXPERIMENTS: PUBLIC HEALTH ENGINEERING LABORATORY

OXYGEN DEMAND IN WATER AND WASTEWATER

The oxygen demand tests are indirect measures of the total organic carbon by determining its
oxidisability, either chemically or biologically. All are gross measures of the total concentration
of organic substances and are usually dominated by the concentration of natural organics.
These tests give an indication of the oxygen required to degrade biochemically and chemically
any organic matter in a water, as well as the oxygen needed to oxidise inorganic materials, such
as sulphides. The tests provide an empirical comparison of the relative oxygen requirements of
surface waters, wastewaters, and effluents. For example, if a sewage effluent with high contents
of of organics is discharged into a stream, the oxygen required by biological organisms to break
down the organic matter in the effluent is taken from the overall oxygen content of the receiving
water. This depletion could potentially destroy fish and plant life.
Because of the importance of estimating the oxygen-demanding potential of wastewaters, several
techniques have been developed over the years. Among the most commonly utilized procedures
are the biochemical oxygen demand (BOD) and chemical oxygen demand (COD) tests.
The BOD test is a batch reactor procedure in which microbes are typically allowed to degrade
organic matter in the sample for a five-day period (BOD5) at 20C. Through noting the volume
of sample placed in the reactor (or BOD bottle) and determining the amount of oxygen used in
the bottle over the five-day period, the BOD of the waste may be calculated.
In the COD test, a wastewater sample is placed in a flask containing chromic acid (dichromate
ions and sulfuric acid), a strong oxidizing solution. After refluxing the sample-oxidant mixture
on a burner for 2 hours, the mixture is removed and the amount of dichromate remaining in the
flask is determined through a redox titration. The amount of dichromate depleted during the test
is proportional to the COD of the sample.
1. BIOCHEMICAL OXYGEN DEMAND
OBJECTIVES
To determine the BoD of a lab sample of Wastewater
INTRODUCTION
Biochemical oxygen demand (BOD) is defined as the amount of oxygen required by
microorganisms, mostly bacteria, to break down decomposable organic matter under aerobic
conditions.
BOD directly affects the amount of dissolved oxygen in running waters. The greater the BOD is,
the more rapidly oxygen is depleted in the system. This means less oxygen is available to higher

forms of aquatic life. The consequences of high BOD are the same as those for low dissolved
oxygen: aquatic organisms become stressed, suffocate, and die. Sources of BOD include leaves
and wood debris; dead plants and animals; animal manure; effluents from pulp and paper mills,
wastewater treatment plants, feedlots, and food-processing plants; failing septic systems; and
urban stormwater runoff.
The Bod Test is meant to measure the dissolved oxygen consumed by bacteria and other
microbial life while the organic substances are present in solution.
This is done under ideal conditions of 200 C in the dark for a specified period of time, normally
adopted as five days.
The reduction of dissolved oxygen concentration in the sample is a measure of the BOD
expressed in mg/l.
The actual environmental conditions in the stream including temperature, biological population,
water movement, light conditions and oxygen concentration can only be simulated but not
accurately reproduced in the laboratory and the results must therefore take into consideration
these variations when related to oxygen demands of natural waters.
Most samples require dilution. If the source of sample is known, its BOD can roughly be
estimated. Accurate results may require more than two dilutions.
A blank sample of distilled water is normally treated and incubated in the same way as the rest of
the samples as a control.
Methods of Testing
The measurements of the dissolved oxygen can be carried out by the Winkler method or with a
Dissolved Oxygen Probe. The Winkler Method was used.
The Winkler method involves filling a sample bottle completely with water (no air is left to bias
the test). The dissolved oxygen is then "fixed" using a series of reagents that form an acid
compound that is titrated. Titration involves the drop-by-drop addition of a reagent that
neutralizes the acid compound and causes a change in the color of the solution. The point at
which the color changes is the "endpoint" and is equivalent to the amount of oxygen dissolved in
the sample
A dissolved oxygen probe is an electronic device that converts signals from a probe that is placed
in the water into units of DO in milligrams per liter. Most meters and probes also measure
temperature. The probe is filled with a salt solution and has a selectively permeable membrane
that allows DO to pass from the stream water into the salt solution. The DO that has diffused into
the salt solution changes the electric potential of the salt solution and this change is sent by

electric cable to the meter, which converts the signal to milligrams per liter on a scale that the
volunteer can read.
APPARATUS AND REAGENTS
Reagents for dilution water:
i.
ii.
iii.
iv.

Phosphate buffer solution

Ferric Solution
Magnesium Sulphate solution and
Calcium chloride solution.

Reagents for DO determination by AZIDE modification of winkler method

i.
ii.
iii.
iv.

Manganese sulphate solution

Concentrated Sulphuric acid
Starch indicator solution and
Standard sodium thiosulphate solution, 0.025 N

Apparatus
Beakers, Burrettes, Pipettes and air suckers, BoD bottles, Incubator
PROCEDURE
A waste water sample is used for the BoD Test.
Dilution water was prepared by adding the following reagents to 6 litres of well aerated water :
a)
b)
c)
d)

6ml of phosphate buffer solution

6ml of ferric chloride solution
6ml of magnesium phosphate solution and
6ml of calcium chloride solution.

The mixture was mixed well and stored under well aerated conditions in an aspirator bottle.
Diluted solutions of the sample (1:10, 1:25; 150 and 1:100) were made by mixing the waste
water sample and the dilution water. Using pipettes, the computed volume of sample was poured
into 280 mm BOD bottles and topped up with the dilution water to make the desired ratios. Care
was taken to ensure no overflowing and the bottle carefully stoppered without trapping any air
bubbles. A blank sample was then fully filled with dilution water alone and stoppered carefully
as for the other samples. 5 sets of each test sample were made and the bottles well labelled
according to the dilution factors used.
The dissolved oxygen content at the beginning of the experiment was then determined for each
of the five sets of bottles. This was done by adding, in quick succession, 2 ml of manganous

sulphate solution followed by 2 ml of alkali azide iodide reagent with the tip of the pipette well
below the water level in the bottle. Ensuring no air bubbles were trapped, the bottles were
stoppered and the contents well mixed by inverting each bottle several times. The precipitates
were then allowed to settle and further mixing done.
2 ml of conc. sulphuric acid were then added carefully using a pipette and air sucker, and the
contents further mixed until all the precipitate had dissolved.
203 ml of the contents were then measured and transferred to an Erlenmeyer flask where it was
titrated against standard sodium thiosulphate solution till the colour of solution just changed to
yellow. 1ml of starch indicator was then added turning the solution to blue; and titration
continued until the blue colour disappeared.
The remaining bottles were incubated at 20 C for five days and the dissolved oxygen determined
as outlined above after 5 days.
RESULTS
DO Determination azide modification
The 203 ml of the solution taken for titration corresponds to 200 ml of the original sample.
Because 1ml of 0.025N Sodium Thiosulphate titrant is equivalent to 0.200 mg DO, each ml of
Sodium Thiosulphate titrant used is equivalent to 1mg/l DO when a volume equal to 200 ml of
original sample is titrated. Therefore
DO mg/l = ml of titrant used under above conditions.
BOD RESULTS
READINGS

Blank

1:10

1:25

1:50

1:100

Final Reading

33.0

6.4

12.5

19

25.6

Initial Reading

25.7

0.5

6.4

12.5

19

Dissolved Oxygen after 5 days

7.3

5.9

6.1

6.5

6.6

Final Reading

29.4

36.2

6.9

7.1

7.2

Initial Reading

22

29.4

Dissolved Oxygen at start of exp

7.4

6.8

6.9

7.1

7.2

READINGS AFTER 5 DAYS

INITIAL READINGS 0 DAYS

CALCULATIONS
12

BoD, mg/l = where

D1, D2 is dissolved oxygen of sample immediately after preparation and after 5days respectively;
P is the fraction of wastewater sample volume to total combined volume (dilution).

SAMPLE

Dilution

DO used

B0D5 = DO/ Dilution

Blank

0.1

0.1

1:10

0.1

0.9

1:25

0.04

0.8

20

1:50

0.02

0.6

30

1:100

0.01

0.6

60

Reported BOD
The average less the blank BOD will be reported:
BOD=

9+20+30+60
4

0.1 = 29.75-0.1 =29.65mg/l

Therefore, BOD5 of sample at 200 C = 30mg/l

DISCUSSION
BOD is the difference between the initial dissolved oxygen in a sample and the dissolved oxygen
in a duplicate sample after a period of five days, expressed in milligrams per litre, divided by the
decimal fraction of the sample used, when examined in accordance with Standard Methods for
the Examination of Water and Wastewater
12
BoD, mg/l = where
D1, D2 is dissolved oxygen of sample immediately after preparation and after 5days respectively;
P is the fraction of wastewater sample volume to total combined volume.
This characteristic of surface water, sewage, sewage effluent, polluted water, industrial waste, or
other wastewater is the amount of dissolved oxygen in milligrams per liter required during
stabilization of the decomposable organic matter by aerobic bacterial action. BOD directly
affects the amount of dissolved oxygen in running waters. The greater the BOD is, the more
rapidly oxygen is depleted in the system. This means less oxygen is available to higher forms of
aquatic life. The consequences of high BOD are aquatic organisms become stressed, suffocate,
and die. Sources of BOD include leaves and wood debris; dead plants and animals; animal
manure; effluents from pulp and paper mills, wastewater treatment plants, feedlots, and foodprocessing plants; failing septic systems; and urban stormwater runoff.

Theoretically, Biochemical oxidation takes an infinite time to go to completion because the rate
of oxidation is assumed to be proportional to the amount of organic matter remaining. Within a
20-day period, the oxidation of the organic matter is about 90 to 99 percent complete, and in 5
day period used for the BoD test, oxidation is about 60-70 percent complete.
If sufficient oxygen is available, the aerobic biological decomposition of organic waste will
continue until all the waste constituents are consumed. During this process, a portion of waste is
oxidized to end products to obtain energy for cell maintenance and the synthesis of new cell
tissue. When all organic matter is used up, the cells begin to consume their own cell tissue to
obtain energy and die off.
The test is affected by factors such as temperatures, type of organic waste, velocity of stream
waters and volumes of the waters.
In the winkler test, Azide modification is done to eliminate inerferences due to nitrite-nitrogen
and ferrous iron effects in the test.
Importance of the BoD test
The BOD test is widely used to determine the pollutional strength of domestic and industrial
wastes in terms of the oxygen that they will require after being discharged into natural water
systems.
In design, BoD is used to determine the sizes of wastewater facilities, like ponds, required to
ensure complete natural organic breakdown of influents before the discharge of effluents.
Engineers have also used BOD tests to measure efficiency of some treatment processes and to
determine compliance with wastewater discharge permits.
Limitations of the test
BoD test has some limitations which have led to preference of other tests in some cases.
For credible results, a high concentration of active, acclimated seed bacteria is required. To
protect the biological microorganisms during toxic waste tests, the waste requires pretreatment
and the effects of nitrifying bacteria must also be reduced.
The other drawback is that only the content of biodegradable organic matter is measured and the
test has no stoichiometric validity after the soluble organic matter present is used up. Further, the
test is very slow and requires well-conditioned procedures for valid results. Even after this
period, the 5 day test period may or may not correspond to the point where all the organic matter
has been used up, making it difficult to have deterministic values of the actual content of organic
matter in a wastewater sample.

CONCLUSION
The BOD of the Waste water sample was found to be 30mg/l.

2. CHEMICAL OXYGEN DEMAND

Objective:
To determine the Chemical oxygen demand of a wastewater sample
INTRODUCTION
An alternative test used to determine the oxygen consumption potential of a wastewater sample
is the COD test.

The chemical oxygen demand (COD) test is a measure of the quantity of oxygen
required to oxidize the organic matter in a waste water sample under specific conditions of
oxidizing agent, temperature and time.
During the determination of COD, organic matter is converted to carbon dioxide and water,
amino nitrogen to ammonia nitrogen and organic nitrogen in higher oxidation states to nitrates
regardless of the biological degradability of the substances. For example glucose (biologically
degradable) and lignin (biologically resistant) are both oxidised completely. As a result COD
values should be greater than BOD values and may be much greater when significant amounts of
biologically resistant organic matter is present.
Potassium dichromate has been found to be the most practical oxidizing agent of all. Organic
substances in the sample are oxidized by potassium dichromate in sulphuric acid solution at
boiling temperature under reflux condensers. Silver Sulphate is used as a catalyst (for oxidation
of straight chain alcohols and acids) and mercuric sulphate is added to remove chloride
interference (which would give rise to an inorganic COD)
The excess dichromate is titrated with standard ferrous ammonium sulphate using
orthophenanthroline ferrous complex as an indicator.

APPARATUS AND REAGENTS

Reagents:
i.
ii.
iii.
iv.
v.

Distilled water
Standard Potassium dichromate solution (0.025N)
Sulphuric acid concentration reagent containing silver sulphate
Standard ferrous ammonium sulphate (0.025N)
Powdered mercuric sulphate

vi.

Phenanthroline ferrous sulphate (ferroin indicator solution)

Apparatus:
i.
ii.
iii.

Reflux apparatus with ground glass joint

250ml Erlenmeyer flask with ground glass joints
Glass beads

iv.

Pipettes

PROCEDURE
To a 250 ml Erlenmeyer flask the following reagents were added:
a) 0.4g solid mercuric sulphate
b) 20.0 ml of sample
c) 10.0 ml of 0.025N potassium dichromate and
d) a few glass beads
The above was repeated using 20.0 ml of distilled water sample as a blank specimen.
The flask was fitted to the condenser system making sure the ground-glass joint was snug.
Cooling water was then passed through the condenser.
30 ml of silver sulphate concentrated sulphuric acid solution was added to the flask through the
open air of the condenser. The contents of the flask were then mixed by swirling by adding the
acid.
The heaters were switched on and refluxed for two hours then switched off. The condensers were
rinsed inside with distilled water, cooled and the flask carefully removed from the heater.
The content of each flask was then diluted with distilled water to about 150 ml while mixing. 2-3
drops of Ferroin indicator solution was added to the flask. The contents of the flask were then
titrated with standard ferrous ammonium sulphate solution of 0.1N strength to an end point noted
with a change of colour from blue-green to reddish-brown.
STANDARDIZATION OF FERROUS AMMONIUM SULPHATE SOLUTION
10.0 ml of 0.25N standard potassium dichromate solution was diluted to about 100 ml in a 250
ml Erlenmeyer flask. 30 ml of concentrated sulphuric acid was then added, mixed thoroughly
and allowed to cool.

The mixture was then titrated with ferrous ammonium sulphate solution from the burette using 23 drops of ferrous as indicator. These results recorded were used t compute and verify the
normality of ferrous ammonium sulphate.
RESULTS
1. Chemical Oxygen Demand Results
initial pipette reading for sample =0 ml
Final pipette reading for sample= 23.0ml
Volume used therefore = 23.0 ml
Initial pipette reading for sample =23 ml
Final pipette reading for sample= 47.3 ml
Volume used therefore = 24.3 ml
2. Ferrous Ammonium Sulphate Standadization
Initial pipette reading for Ferrous Ammonium Sulphate = 9.00ml
Final pipette reading for Ferrous Ammonium Sulphate = 33.9 ml
Volume used therefore

= 24.9 ml

CALCULATIONS

1. Standardisation of Ferrous Ammonium Sulphate

0.25

Normality =
=

0.25 =10
24.9

= 0.1 N
This satisfies the requirements for the test.

2. Chemical Oxygen Demand for sample

()8000

Mg/l COD =

Where a= ml of titrant used for blank

b= ml of titrant used for sample
N=Normality of titrant
Mg/l COD =

24.723
20

x 0.1 x 8000

= 68 mg/ l
DISCUSSION
COD is a measure of the amount of oxygen, in milligrams per liter, chemically (rather than
biologically) consumed under specific conditions in the oxidation of organic and oxidizable
inorganic materials in water or wastewater.
It has been found that BOD is equal to about 67% COD for any given waste water sample.
The COD is substantially higher than the COD for the following reasons:
a. Many organic substances which are difficult to oxidize biologically such as lignin can be
oxidized chemically
b. Inorganic substances that are oxidized by dichromate increase the apparent organic
matter content of the sample
c. Certain organic substances may be toxic to microorganisms used in the COD test
From an operational standpoint, one of the advantages of the COD test is that it can be completed
in about 3 hours compared to the 5 day period required for a BOD test.
Importance of COD test in Public Health engineering
The COD test is used extensively in the analysis of industrial wastes. Results may be obtained
within a relatively short time(3hrs) In conjunction with the BOD test, the COD test is helpful in
indications toxic conditions and the presence of biologically resistant organic substances. The
test is widely used in the operation of treatment facilities because of the speed with which results
can be obtained.

CONCLUSION
COD of the waste water sample was found to be 68 mg/l