EXPERIMENT NO.

DETERMINATION OF BIO-CHEMICAL
OXYGEN DEMAND OF WASTE WATER
(BOD)

EXPRIMENT NO.
DETERMINATION OF BIO-CHEMICAL
OXYGEN DEMAND OF WASTE WATER (BOD)
1.CONCEPT AND SIGNIFICANCE
BOD is the amount of oxygen required for the biological dcomposition of
the organic matter in the aerobic condition. This test is very important in analysis of
sewage and industrial wastes, as it indicator the amount of decomposable organic
matter in the waste. Greater the BOD value more is the nuisance potential, or the
strength of liquid waste.
Tests for BOD are usually made at a temp. of 20 C and for 5 days. The test
is based on determined of dissolved oxygen, prior to and following a 5 day period of
incubation at 20 C. The sample of sewage is diluted with dilution water. As the BOD
of the sample in titration depend upon the dilution used, several dilution of the
sample should be prepared and tested. The dissolved oxygen measurement of the
dilution water before incubation (called blank)and if the diluted sample after
incubation at 20C.

2. PRINCIPLE
BOD is quantity of oxygen utilized by a mixed population of microorganisms in aerobic oxidation of organic matter at 20 C.The dilution water
containing phosphate buffer, manganese sulphate. Calcium chloride and ferric
chloride is satured with dissolved oxygen.
The waste water supplies the organic matter (food) and the dilution waste
supplies dissolved oxygen. Depletion of dissolved oxygen in the test bottle is directly
related o the amount of degradable organic matter. However the degree of
reproducibility of BOD cannot be precisely defined on account of variations that
occur in decomposition of organic matter.

3. APPARATUS
A) Bod incubator
B) Burette,pipette, measuring cylinder etc.

4. REAGEN
A) 0.025 N sodium thio-sulphate solution
B) Conc.H2SO4
C) Manganess sulphate solution

D) Alkaline iodide azide reagent

E) Starch indicator
F) Phosphate buffer (all as per expt no.9)
G) Ferric chloride
H) Calcium chloride solution

5. PROCEDURE
A) take pure water in a glass container (about 5 litres ) and bubble compressed
air for 2 days to attain saturation.
B) Add 1ml each of manganese sulphate, phosphate buffer, ferric chloride and
calcium chloride solution for each litre of distilled water.
C) If the waste water is not expected to contain sufficient bacteria population,
add seed to dilution water (generally 2 ml of settled sewage for 1000ml of
water.)
D) Neutralize the sample of 7 Ph.
E) The dilution of the sample are made such that about 50% depletion of DO
take place and residual DO after incubation for 5 days is not less than 1
mg/lit.
F) The following dilution are suggested.
1) 0.1% to 1%
2) 1% to 5%
3) 5% to 25%

Strong wastes
Raw sewage
Treated effluent

G) Take 6 BOD bottles, prepare 2 bottles blank, for determination of initial Do

H) Add in 2 bottles necessary diluted sample. Keep 4 bottles in incubator at 20
C for 5 days.
I) Take 1 bottles blank and one sample bottle and determine its initial DO
contents by adding manganese sulphate, alkaline iodide azide agent and
conc. H2SO4 and titrate. DO same with 0.025 N sodium thio-sulphate
solution using starch as indicator. Note the burette reading.
J) After 5 days determine DO in the incubator bottles(blank and with sample)
in the similar way.
DO = DO of sample on-oth day
D1 = DO of sample on-5th day
CO = DO of blank on 0th day

C1 = DO of blank on on 5th day

Then BOD IN = (DO-D1) (CO C1) . percentage dilution
6. OBSERVATION TABLE AND SAMPLE
CALCULATION
SR.
BOTTLE DILUTION
NO.
NO.&
FACTOR
DO
COTENT
0TH DAY
1.
2.
3.
4.
5.
6.

D1
5TH DAY

CO
0TH DAY

C1
5TH DAY

AVE

AVE