Lab W5 BOD

EAB 3209 (Agricultural Waste Management)
LAB W5: Determination Of Biochemical Oxygen Demand (BOD)
Introduction:
The BOD test is used to measure waste loads to treatment plants, determine plant efficiency (in
terms of BOD removal), and control plant processes. It is also used to determine the effects of
discharges on receiving waters. A major disadvantage of the BOD test is the amount of time (5
days) required to obtain the results.
When a measurement is made of all oxygen consuming materials in a sample, the result is
termed “Total Biochemical Oxygen Demand” (TBOD), or often just simply “Biochemical Oxygen
Demand” (BOD). Because the test is performed over a five day period, it is often referred to as a
“Five Day BOD”, or a BOD5.
In many biological treatment plants, the facility effluent contains large numbers of nitrifying
organisms which are developed during the treatment process. These organisms can exert an
oxygen demand as they convert nitrogenous compounds (ammonia and organic nitrogen) to
more stable forms (nitrites and nitrates). At least part of this oxygen demand is normally
measured in a five day BOD.
Sometimes it is advantageous to measure just the oxygen demand exerted by organic

(carbonaceous) compounds, excluding the oxygen demand exerted by the nitrogenous
compounds. To accomplish this, the nitrifying organisms can be inhibited from using oxygen by
the addition of a nitrification inhibitor to the samples. The result is termed “Carbonaceous
Biochemical Oxygen Demand”, or CBOD.
Procedures:
1.0 Fast method
A. Dilution water preparation [BOD Nutrient Buffer Pillows]

1. Aerate 1L of distilled water(A) for 3-5 hours.
2. Add 1 pillow of BOD vial into the solution A and stir.
3. Measure the initial DO reading (D1) with YSI probe.
Sample Solution
Preparation of dilution water: Add 1mL each of phosphate buffer, magnesium sulfate,
calcium chloride, ferric chloride solution into 1L volumetric flask. Add distilled water to 1L and
stir well. [Replace with Fast Method (use the solution prepared in part A)]
1. Add 1mL wastewater sample into a 500mL beaker.

2. Add dilution water up to 300mL into the same beaker.
3. Measure DO for each sample by using Dissolved Oxygen Meter/YSI Probe.
4. Record initial DO (D1)
5. Put all prepared samples and control in 300mL-incubation bottle.
6. Fill up the incubation bottle with some distilled water for air seal before capping.
7. Put all the bottles in BOD Incubator for 5 days. Set the temperature at 20°C.
8. Measure final DO value (D2) after 5 days.
Calculations:
BOD5, mg/L = (D1 – D2) / P
Where;
D1 = DO value in initial sample

D2 = DO value in final sample
P = Decimal volumetric fraction of sample used
Or;
WE USE THIS
BOD5, mg/L = (D1 – D2) x Dilution factor
Dilution factor = Bottle volume (300mL) / Sample volume (1mL)

****[1ml sample:299ml dilution water]
Example:
Initial DO = 8.2 mg/L
Final DO = 4.4 mg/L
Sample size = 5 mL
BOD mg/L = [(8.2 - 4.4) x 300]/5 = (3.8 x 300)/5 = 1140/5 = 228 mg/L
2.0 Titration method
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 Day 0 and 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

P
Where P = volumetric fraction of sample to total BOD bottle volume
A. Determination of DO
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.

Quiz
1. What does a BOD test measure?
2. What can the BOD test be used for?
3. What are the types of samples and maximum sample holding time?
4. What reagents are required for the BOD5 and/or CBOD5 test?
5. What equipment, apparatus, or instrumentation is required for the BOD5 and/or

for CBOD5 test?
6. If the expected BOD of a sample is in the range 25 mg/L to 75 mg/L, what would
be the minimum and maximum sample volumes to use for the sample dilution?

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EAB 3209 (Agricultural Waste Management)

LAB W5: Determination Of Biochemical Oxygen Demand (BOD)

Sometimes it is advantageous to measure just the oxygen demand exerted by organic

1.0 Fast method

A. Dilution water preparation [BOD Nutrient Buffer Pillows]

1. Add 1mL wastewater sample into a 500mL beaker.

BOD5, mg/L = (D1 – D2) / P

D1 = DO value in initial sample

BOD5, mg/L = (D1 – D2) x Dilution factor

Dilution factor = Bottle volume (300mL) / Sample volume (1mL)

Initial DO = 8.2 mg/L

Final DO = 4.4 mg/L

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

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

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

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

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

1. What does a BOD test measure?

2. What can the BOD test be used for?

5. What equipment, apparatus, or instrumentation is required for the BOD5 and/or

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