12.

BIOCHEMICAL OXYGEN DEMAND (BOD)

Objective:

To determine biochemical oxygen demand (bod) of given water/waste water sample.

Introduction:

Biochemical Oxygen Demand (BOD) determines the amount of dissolved oxygen needed by
aerobic organisms to break down organic materials in a water sample at a certain temperature
over a specific period. BOD is the oxygen required for the biological decomposition of dissolved
organic matter under standard conditions at a standardized time (usually 5 days) and temperature
(20°C).

The test measures molecular oxygen used for the biochemical degradation of organic material
(carbonaceous demand) and the oxygen used to oxidize inorganic materials like sulfides and
ferrous ion. It may also measure oxygen used to oxidize reduced forms of nitrogen (nitrogenous
demand).

Environmental Significance:

BOD indicates the biodegradability of a sample and waste strength, helping to measure pollution
levels and the efficiency of treatment plants.

BOD is used in organic waste discharge assessment, it helps calculate the effect on oxygen
resources of receiving waters and is critical for designing wastewater treatment plants.

Guidelines:

 Drinking water: BOD should be less than 1 mg/L.

 Effluent to natural bodies: BOD should be less than 30 mg/L.
 BOD of typical domestic sewage: around 200 mg/L.
 Regulatory standards in Bangladesh specify a drinking water BOD of 0.2 mg/L and an
allowable effluent BOD of 50-250 mg/L depending on the discharge point.

Principle:

The sample is incubated at 20°C for 5 days in an airtight bottle. Dissolved Oxygen (DO) is
measured before and after incubation, and BOD is calculated from the difference between initial
and final DO. Due to high oxygen demand, samples often need to be diluted to keep DO above
zero. If DO drops to zero, the test is invalid.

The five-day BOD of a diluted sample is given by,

BOD5 = [DOi- DOf] × D.F. …………………………. (1)

In some cases, it becomes necessary to seed the dilution water with microorganisms to ensure
that there is an adequate bacterial population to carry out the biodegradation. In such cases, two
sets of BOD bottles must be prepared, one for just the seeded dilution water (called the "blank")
and the other for the mixture of wastewater and dilution wader. The changes in DO in both are
measured. The oxygen demand of waste water (BODw) is then determined from the following
relationship:

BODm× Vm= BODw × Vw + BODd × Vd ……………………… (2)

Where,

BODm, is the BOD of the mixture of wastewater and dilution water

BODd is the BOD of the dilution water alone;

Vw and Vd are the volumes of wastewater and dilution water respectively in the mixture and
Vm= Vw+ V

Sample Handling:

Samples should be analyzed within 2 hours of collection. If this is not possible, keep samples at
4°C and analyze within 6 hours. Do not allow samples to freeze or open the bottle before
analysis.

Apparatus:

1. BOD bottle
2. Beaker (250 ml)
3. Measuring cylinder
4. Dropper
5. Stirrer

Reagents:

1. Manganous sulfate solution

2. Alkaline potassium iodide solution
3. 0.025N sodium thiosulfate
4. Starch solution (indicator)
5. Concentrated sulfuric acid

Procedure:

Sample Preparation:

1. Fill two BOD bottles with the sample (or diluted sample), ensuring that the bottles are
completely filled.
 2. Immediately determine the initial dissolved oxygen (DO) concentration (DOi) in one
bottle after filling.
3. Store the other bottle in a dark location at 20°C. After the designated incubation period
(typically 5 days), determine the final dissolved oxygen concentration (DOf) in the
sample.

Measurement Procedure:

1. Using a pipette, add 1 mL of manganous sulfate solution to the BOD bottle, ensuring the
pipette tip is just below the surface of the water.
2. Similarly, add 1 mL of alkaline potassium iodide solution to the BOD bottle in the same
manner.
3. Insert the stopper into the bottle and mix the contents by inverting the bottle several
times.
4. Allow the precipitates to settle halfway and then mix the solution again.
5. Allow the precipitates to settle halfway once more.
6. Add 1 mL of concentrated sulfuric acid to the BOD bottle. Immediately insert the stopper
and mix by inverting the bottle.
7. Let the solution stand for at least 5 minutes to ensure complete reaction and settling.

Titration:

1. Withdraw 100 mL of the solution into an Erlenmeyer flask.

2. Add 0.025N sodium thiosulfate solution drop by drop from a burette until the yellow
color of the solution nearly disappears.
3. Add approximately 1 mL of starch solution to the flask and continue adding sodium
thiosulfate drop by drop until the blue color just disappears.
4. Record the volume of sodium thiosulfate solution used (ignore any return of the blue
color).

Results and Calculations:

Sample Dilution Factor DOi (mg) DOf (mg) BOD5 (mg/L)

Control Blank - 7.2 7.1 -
Sa 10 6.9 0.5 64
Sb 25 7.0 1.9 127.5
Sc 50 7.1 5.0 105.0
Sd 100 7.2 5.4 180.0

The five-day BOD of a diluted sample is given by:

BOD5= (DOi−DOf) ×D.F.

Where:
  DOi: Initial Dissolved Oxygen
 DOf: Final Dissolved Oxygen
 D.F. Dilution Factor

Control Blank:

BOD5= (DOi−DOf) ×D.F.

The blank sample has no dilution factor, so no meaningful BOD5 calculation is made.

Sample Sa:

BOD5= (6.9−0.5) ×10

BOD5=6.4×10=64.0 mg/L

Sample Sb:

BOD5= (7.0−1.9) ×25

BOD5=5.1×25=127.5 mg/L

Sample Sc:

BOD5= (7.1−5.0) ×50

BOD5=2.1×50=105.0 mg/L

Sample Sd:

BOD5= (7.2−5.4) ×100

BOD5=1.8×100=180.0 mg/L

Discussion:

The experiment measured BOD5 of water samples with varying dilution factors to assess organic
pollution levels. The control showed minimal DO reduction, indicating low biological activity.
Higher BOD5 values in diluted samples (64.0–180.0 mg/L) reflect significant organic pollution,
with the highest values suggesting heavily contaminated water. These levels exceed safe limits
for drinking water (<1 mg/L) and effluent discharge (<30 mg/L), highlighting the need for better
wastewater treatment. Proper dilution and seeding ensured accurate results, but the samples' high
oxygen demand underscores the severity of pollution. Regular monitoring and advanced
treatment methods are essential to reduce environmental impact.

Conclusion:
The results indicate that the tested water samples contain significant levels of organic pollution,
as evidenced by high BOD5 values ranging from 64.0 to 180.0 mg/L. Such levels suggest
untreated or inadequately treated wastewater, making the water unsuitable for drinking or safe
discharge into natural bodies.

11. DETERMINING CHEMICAL OXYGEN DEMAND (COD)

Objective:

To determine the Chemical Oxygen Demand (COD) of given sample.

Principle:

Potassium permanganate is used as the oxidizing agent in the COD test. It selectively oxidizes
carbonaceous organic matter in the sample. An excess of the oxidizing agent is used, and the
remaining excess is measured by titration with a reducing agent like ammonium oxalate.

Apparatus:

1. Beaker (250 mL)

2. Dropper
3. Stirrer

Reagents:

1. Diluted sulfuric acid solution

2. Standard potassium permanganate solution
3. Standard Ammonium Oxalate solution

Procedure:

1. Pipette 100 mL of the sample into a 250 mL Erlenmeyer flask.

2. Add 10 mL of diluted sulfuric acid and 10 mL of standard KMn04 solution.
3. Heat the flask in a boiling water bath for exactly 30 minutes, keeping the water in the
bath above the level of the solution in the flask. The heating enhances the rate of
oxidation reaction in the flask.
4. If the solution becomes faintly colored, it means that most of the potassium permanganate
has been utilized in the oxidation of organic matter. In such a case, repeat the above using
a smaller sample diluted to 100 mL with distilled water.
 5. After 30 minutes in the water bath, add 10 mL of standard ammonium oxalate
[(NH4)2C204] solution into the flask. This 10 mL ammonium oxalate, which is a
reducing agent, is just equivalent to the 10 mL potassium permanganate (oxidizing agent)
added earlier. The excess of reducing agent [(NH4)2C204] now remaining in the flask is
just equivalent to the amount of the oxidizing agent (KMn04) used in the oxidation of
organic matter.
6. The quantity of ammonium oxalate remaining in the flask is now determined by titration
with standard potassium permanganate. Titrate the content of the flask while hot with
standard potassium permanganate to the first pink coloration. Record the mL of
potassium permanganate used.

Results and Calculations:

20.4 ml of potassium permanganate used.

Discussion:

In this experiment, we determined the Chemical Oxygen Demand (COD) of the given sample
using potassium permanganate as the oxidizing agent under acidic conditions. COD is an
important parameter for evaluating the amount of organic pollutants in water by measuring the
total oxygen required to oxidize organic matter to carbon dioxide and water.

The COD test offers several advantages, including its quick results compared to the Biological
Oxygen Demand (BOD) test, which requires five days. However, it has limitations as it cannot
distinguish between biodegradable and non-biodegradable organic matter. Despite this
limitation, COD remains a widely used indicator in environmental studies, wastewater treatment
plants, and industries such as pulp and paper mills and power plants due to its ability to rapidly
assess organic pollution.

The experimental COD value was determined using 20.4 mL of potassium permanganate, which
was consumed during the oxidation process and found to be 20.4mg/dm3 O2. Upon comparison
with standard guidelines, such as the Bangladesh Environment Conservation Rules (1997),
where drinking water COD is limited to 4.0 mg/L and typical wastewater effluent COD ranges
between 200–400 mg/L, the measured value was analyzed to assess compliance and treatment
efficiency.
Environmental Implications:

The significance of the COD measurement lies in its ability to indicate the level of organic
pollution in water bodies. Deviations from acceptable COD ranges suggest potential
environmental risks, including oxygen depletion in aquatic ecosystems. High COD values in
wastewater effluents may indicate inadequate treatment and can harm aquatic life.

This experiment reinforces the importance of regular COD monitoring in maintaining water
quality and highlights the need for effective treatment methods to minimize the release of
organic pollutants into natural water bodies.

Conclusion:

The Chemical Oxygen Demand (COD) of the given water sample was determined to be 20.4
mg/dm³ O₂. This value indicates the amount of oxygen required to oxidize the organic matter in
the sample. Based on standard guidelines, such as the Bangladesh Environment Conservation
Rules (1997), which recommend a COD of 4.0 mg/L for drinking water, the measured COD
suggests that the sample may not meet the criteria for potable water. However, it falls well below
the typical range for untreated wastewater effluents, which is 200–400 mg/L, indicating
relatively low levels of organic pollution. This result highlights that the water sample is not
heavily polluted and may have undergone some level of treatment.