MODERNA

BOD estimation

Problem:
You are provided with an effluent sample from a cheese factory. Estimate its BOD and
determine whether it can be disposed into a natural water body without further treatment.

Approach:
The amount of oxygen consumed by microorganisms in the breakdown of organic is known as
the biochemical oxygen demand or BOD. Wastewater from sewage treatment plants and food
industries often contains organic materials that are oxidized by microorganisms, hence using
oxygen in the process. Oxygen is measured in its dissolved form as dissolved oxygen (DO). If
more oxygen is consumed than is produced, dissolved oxygen levels decline and some
sensitive animals may get affected or die, thus putting the aquatic ecosystem under stress.
The rate of oxygen consumption in a stream is affected by a number of variables: temperature,
pH, the presence of certain kinds of microorganisms, and the strength of the municipal or
industrial effluent, thus, the type of organic and inorganic material present in the water. BOD
directly affects the amount of dissolved oxygen in water bodies. The greater the BOD the more
rapidly is the oxygen being depleted in the water body. This means less oxygen available for
higher forms of aquatic life. The consequences of high BOD are the same as those for low
dissolved oxygen- aquatic organisms become stressed, suffocated, and die. BOD reduction is
used as a criterion to judge the effectiveness of wastewater treatment.

BOD assay is a procedure that measures the dissolved oxygen consumed by bacteria from the
decomposition of organic matter in a given sample. The test does not determine the total
amount of oxygen demand present, it measures only the organic matter that can be biologically
oxidized by aerobic bacteria. Non-degradable substances such as plastics, ribbons and
thermocols are not considered in the estimation of BOD.

This test is carried out by appropriately diluting the given effluent sample in two BOD bottles.
The oxygen level in one of the bottles is recorded on the first day, while the dissolved oxygen
content in the second bottle is measured after incubation at 20 deg for 5 days. The difference in
the dissolved oxygen content of both the samples is used in the formula for the calculation of
the biochemical oxygen demand. For estimation of dissolved oxygen by Winkler method, an
iodometric titration of the effluent sample with sodium thiosulphate is carried out. The effluent
sample is reacted with manganese sulfate and alkaline iodide to form a manganic hydroxide
precipitate. Concentrated sulphuric acid is added to dissolve the precipitate. The added iodide
reduces the manganese ions and in turn liberates iodine. Titration involves the drop-by-drop
addition of N/80 sodium thiosulphate reagent with liberated iodine using starch as indicator that
causes a change in the colour of the solution. The point at which the colour changes from blue
to colourless is the endpoint and is equivalent to the amount of oxygen dissolved in the sample.
Requirements:
Sr. No. Particulars Volume Number

Sample/Reagents

1. Effluent sample from cheese factory 10 mL

2. Aerated distilled water 1000 mL

3. Starch indicator

4. Sodium thiosulfate (Na2S2O3) N/80 25 mL

normality

5. Magnesium sulfate (MnSO4) 4 mL

6. Alkaline iodide solution 4 mL

7. Concentrated Sulphuric acid (H2SO4) 4 mL

Glassware

1. Sterile pipette 10 mL 2

2. Conical flask 250 mL, 1L 2

BOD bottles 300 mL 2

Instruments

1. Incubator 1

Miscellaneous

1. Measuring cylinder 100 mL 1

2. Dropper 1

3. Aeration tubes

Procedure:
● Take 800 mL of 1:100 diluted effluent sample in a 1-litre flask.
● For sufficient amount of dissolved oxygen, aerate the sample by passing air bubbles into
the sample for 2 hours.
● Fill two 300mL BOD bottles completely up to the brim with the aerated sample. Make
sure no space is left and no air is trapped which could give inaccurate results.
● Label one1 of the bottles as DO1 and use it immediately to determine the amount of
dissolved oxygen present initially
● Label the second bottle as DO5 and incubate it for 5 days with a water seal.
● At the end of 5 days of incubation, determine the amount of dissolved oxygen present.
 ● Find the difference in the dissolved oxygen contents of both the BOD bottles and apply it
to the formula to calculate the biochemical oxygen demand value of the effluent sample.

2. Estimation of dissolved oxygen (DO) content:

● Add 2 mL each of MnSO4 and alkaline iodide solution to both the BOD bottles
● During the addition of reagents, make sure to keep the tip of the pipette well below the
surface of the sample. This is to ensure no extra aeration is incorporated into one of the
BOD bottles
● Replace the stopper and mix thoroughly by inversion for 1 minute.
● A brown manganic hydroxide MnO(OH)2 precipitate is formed and is allowed to settle
down till two-thirds of the solution is clear.
● 2 mL of concentrated H2SO4 is added to dissolve the precipitate and the solution turns
orange-ish in colour.

3. For Titration:

● Take 100 ml of the solution from the BOD bottle into a conical flask.
● Pipette out N/80 Na2S2O3 (Sodium thiosulphate) and add it drop by drop into the flask
till the solution turns straw yellow in colour.
● After the solution has turned straw yellow, we add two drops of starch indicator. The
solution in the flask turns blue in colour
● Titrate the liberated iodine drop by drop against N/80 Na2S2O3 until the solution
reaches the endpoint, that is, until it turns colourless.

Observations:
1. Addition of reagents to effluent sample

Formation of brown The precipitate is allowed to Addition of concentrated

precipitate of MnO(OH)2 on settle down until ⅔ rd of the H2SO4 that dissolves the
addition of MnSO4 and solution appears clear precipitate and gives an
alkaline iodide to effluent orange colour to the solution.
sample
 2. Titration of BOD sample with sodium thiosulphate

The solution is titrated with
N/80 Na2S2O3 until it turns
straw yellow in colour.
The solution turns blue in
colour after addition of two
drops of starch indicator
On further titration with N/80
Na2S2O3, solution reaches
endpoint i.e colourless and
the titration reading is noted
down

3. Titration readings
Sample Dilution Dissolved oxygen content
name factor
Bottle Titration Bottle Titration
number reading number reading

Effluent 100 DO1 10.4 mL DO5 6.2 mL

sample from
a cheese
factory
We obtained a considerably lower titration reading for day 5 i.e 6.2 mL compared to day 1 i.e
10.4 mL

Calculations and Result:

We used N/80 normality sodium thiosulphate
1000 ml of 1N Na2S2O3 = 8g oxygen
1000 ml of N/80 Na2S2O3 = 8*(1/80) = 0.1g of oxygen
Consider ‘x’ is the amount of Na2S2O3 that we used
Therefore,
‘x’ ml of of N/80 Na2S2O3 = 0.1x/1000 g of oxygen
= 10^-4x grams of oxygen
We used 100 ml of sewage sample for titration
Therefore,
100 ml of the effluent sample contains = 10^-4x g of oxygen
1000 ml of effluent sample contains = 10^-3x g of oxygen
x*10^-3 g of oxygen = x mg of oxygen.
The concentration of dissolved oxygen in the sample is equivalent to the mL of titrant used.
Each mL of sodium thiosulphate used in titration is equal to 1 mg of dissolved oxygen.

Therefore,
Titration reading for day 1 bottle = 10.4 mL
= 10.4 mg of dissolved oxygen.
Titration reading for day 5 bottle = 6.2 mL
= 6.2 mg of dissolved oxygen.

Calculation of Dissolved Oxygen content

1000 ml of 1N Na2S2O3 is equivalent to 8 g of O2

D.O. (mg/L) = B.R x N x 8 x 1000

V

B.R: Burette reading

N: Normality of Na2S2O3

V: Volume of sample titrated

BOD = [DO1 – DO5] x D.F

ESTIMATION OF BOD:
DO1 value =10.4 mg
DO5 value = 6.2 mg
Dilution Factor = 100
Formula = (DO1-DO5) * Dilution Factor
= (10.4-6.2) * 100
= 4.2 * 100
= 420 mg/L
Therefore, the BOD value of the given effluent sample is estimated to be 420 mg/L
Conclusion:
We were provided with an effluent sample from a cheese factory and were asked to estimate its
BOD and determine whether it can be disposed into a natural water body without further
treatment. We estimated the BOD level of the effluent sample using Winkler’s method of BOD
estimation. Magnesium sulfate added into the effluent sample resulted in the formation of a
brown coloured precipitate which was dissolved by adding concentrated sulphuric acid. Alkaline
iodide reduces the magnesium ions, in turn, liberating iodine. This solution on titration with
sodium thiosulfate estimates the total amount of dissolved oxygen present in the sample. The
dissolved oxygen content of the effluent sample was estimated on Day 1 and again after 5 days
and the values obtained were compared to estimate the BOD value of the effluent sample.
On calculation, the BOD value that we obtained for our Effluent sample was 420 mg/L
The permissible BOD limit for effluent or sewage wastewater that can be released into inland
water bodies is less than 30 mg/mL. The BOD value of the provided effluent sample is way
greater than the permissible value. The effluent sample still contains a high amount of organic
content and the water treatment system employed is not as effective as it should be. Care must
be taken to ensure that the effluent getting released from the cheese factory meticulously goes
through the primary, secondary and tertiary processes such as trickling filters, oxidation ponds,
fixed film systems, activated sludge processes of wastewater treatment before getting released
into the environment. Release of such untreated effluent water can cause oxygen depletion and
contamination of the receiving water body, thus affecting the aquatic flora and fauna.

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MODERNA

AYESHA ANSARI APPROACH

SHWETA GUPTA REQUIREMENTS

KAREENA PIRDHANANI PROCEDURE

AKSHATA KIRLOSKAR OBSERVATIONS

NEHA MAURYA IMAGES

ASHRAMI MHATRE RESULT AND CALCULATIONS

MAHITA LAVETI CONCLUSION