UNIVERSITI PUTRA MALAYSIA (UPM)

FACULTY OF ENGINEERING
DEPARTMENT OF CIVIL ENGINEERING
WATER AND ENVIRONMENTAL ENGINEERING UNIT

WATER AND WASTEWATER ENGINEERING

Biochemical Oxygen Demand (BOD) and Dissolved Oxygen (DO) Determination

Introduction:

Biochemical Oxygen Demand (BOD) is defined as the requirement or demands for oxygen that matter,
organic and chemical during the decay process or during chemical reactions or BOD is said to be a measure of
the quantity of oxygen (determined by DO content) used by microorganisms (e.g., aerobic bacteria) in the
oxidation of organic matter. In other words, BOD is the measurement of how much stress is being placed on the
dissolved oxygen system of a river or waterway. Any wastes, such as municipal sewage, abattoir discharge, and
some industrial wastes with high BOD, must be treated to remove or lower the BOD before release into the
receiving water. The five-day Biochemical Oxygen Demand (BOD5), a traditional time duration of 5 days is a
standardized assessment of the amount of oxidizable, or respirable, organic matter in water.  It is the amount of
oxygen (in mg/L) required by bacteria to oxidize the organic molecules aerobically. 

Objective: i) To determine the level of BOD5 and DO (using Azide Modification Method) of a sample
given.

Apparatus: 1. Pipette 4. Measuring cylinder

2. Burette with retort stand 5. BOD bottle (300 ml)
3. Conical flask 6. Incubator at 20oC

Reagent and solutions: 1. Manganese sulfate solution (MnSO4)

1. Alkaline – Iodide Azide solution
2. Concentrated sulfuric acid (Conc. H2SO4)
3. Sodium thiosulfate titrant (Na2S2O3)
4. Starch indicator
5. BOD dilution water

Chemical Preparation:

A. BOD Dilution Water

1. Place those nutrients, 1 ml of each Phosphate Buffer, MgSO4, CaCl2 and FeCl3. 6H2O solution per liter
of distilled water.
2. Saturate the dilution water by aerating with organic free filtered air for around 1 hour and cover the
dilution water container before let the content settle down.

B. Manganese Sulfate Solution, MnSO4

1. Dissolve 480 g MnSO4.4H2O, 400 g MnSO4.2H2O or 364 g MnSO4.H2O in distilled water, filter and
dilute to 1 L.

C. Standard Sodium Thiosulfate Titrant, Na 2S2O3

1. Dissolve 6.205 g Na2S2O3.5H2O in 250 ml distilled water

2. Add 1.5 ml 6N NaOH or 0.4 g solid NaOH and dilute to 1000 ml

D. Alkali Iodide Azide reagent

1. Readily purchased

E. Concentrated Sulfuric Acid, H 2SO4

 1. Readily purchased

F. Starch Indicator

1. Dissolve 2 g laboratory grade soluble starch and 0.2 g salicylic acid (preservative) in 100 ml hot
distilled water

Methodology:

A. BOD Test

1. Prepare 12 BOD bottles (300 ml). Allocate 4 bottles each for 2 type of samples and remaining 4
bottles for blank.
2. For each type of sample, divide 2 bottles each for
(i) Day 0 and
(ii) Day 5
Label the bottles.

3. For bottles labeled sample (Day 0 and Day 5), pipette 1 ml sample or diluted sample into the BOD
bottle. For bottles labeled blank, pipette 1 ml distilled water into the BOD bottle.
4. For each BOD bottle, fill till the neck of the BOD bottle with BOD dilution water.
5. Cap all the BOD bottles tightly and ensure that no bubble formed or present in the bottle.
6. For BOD bottle labeled Day 5 (sample and blank), seal the cap together with the neck of the bottle
with parafilm provided. Wrap the whole bottle with aluminum foil provided and incubate in
incubator at 20oC for 5 days.
7. For BOD bottle labeled Day 0 (sample and blank), determine the initial DO content using Azide
Modification Method.
8. Determine the final DO content for sample and blank for Day 5 using Azide Modification Method.

Calculation:

BOD5 (mg/L) = DO Initial - DO Final(5)

P (1ml/300ml)

Where P = volumetric fraction of sample to total BOD bottle volume

B. Determination of DO (If you have Dissolve oxygen meter, no need to apply this method)

1. In each BOD bottle, add 1 ml MnSO4 followed by 1 ml Alkaline- Iodide Azide reagent.
2. Cap the bottle and mix inversely for 5 times.
3. Let the suspension settle fully.
4. Then, add 1 ml Conc. H2SO4 into the bottle.
5. Cap and invert the bottle until the suspension dissolve completely.
6. Take 200 ml sample (or blank) from the BOD bottle and pour into conical flask.
7. Titrate with Na2S2O3 to the pale yellow colour.
8. Add 2 – 3 drop of starch indicator (sample become slightly blue) and continue the titration until
sample become colourless.
9. Record the volume of Na2S2O3 used for each titration.

Calculation:

DO (mg/L) = total volume of Na2S2O3 used for each titration

1 ml of Na2S2O3 titrant = 1 mg/L of DO

Results:

1. Report results in tables and show all the calculations.