DETERMINATION OF

BIOCHEMICAL OXYGEN
DEMAND (BOD)
 AIM

To determine biochemical oxygen

demand in the given water sample with
the stipulations as per IS
Relevant Indian Standard for Dissolved
Oxygen Test :

IS: 3025 (Part 44) - Reaffirmed 2003.

 Introduction
The biochemical oxygen demand is a chemical procedure
for determining the amount of dissolved oxygen needed
by aerobic organisms in a water body to break the organic
materials present in the given water sample at certain
temperature over a specific period of time.

Usually, the time is taken as 5 days and the

temperature is 20°C.
 Introduction
The test measures the molecular oxygen utilized during a
specified incubation period for the biochemical
degradation of organic material and the oxygen used to
oxidize inorganic material such as sulfides and ferrous
ion.
 Significance
1. Principle test to give an idea of the biodegradability of any
sample and strength of the waste.
2. Efficiency of any treatment plant can be judged
3. Allows calculation of the effect of the discharges
4. Any effluent to be discharged into natural bodies of water should
have BOD less than 30 mg/L
5. when BOD value reaches 5 mg/L, the water is doubtful in purity
6. to measure the self-purification capacity of streams
7. measure of the amount of clean diluting water required for the
successful disposal of sewage
 Principle
A known volume of a sample of a waste water, diluted
using specially prepared distilled water, is incubated at20˚
c for 5 days.

DO depletion in the test sample is a measure of a fair

amount (68%) of biodegradable organic matter in the
sample To avoid getting a zero 5-day BOD, a dilution
technique is adopted.
 DO Fixation:-
DO Fixation is done by adding 2ml of manganous sulphates
and 2ml of alkali-iodide-azide.

For best results the sample must be stored below 10˚C during
transits and titrated with in six hours of fixation
 Winkler’s Modified Method :

The Winkler’s Method is the technique used to

measure DO in freshwater structures.

It is used as an indicator of the health of a water

body, where higher DO concentrations are
associated with high production and little
contamination.

This test is performed on-site, as delays between

sample collections and testing may result in a
variation in oxygen content.
 The BOD test is considered a failure for
any particular dilution
a) If the 5 day DO is equal to, or nearly equal to, immediate DO
i.e.. dilution has been too much.

(b) If the 5-day DO is equal to zero or less than 0.5mg/l i.e. dilution
has been too less. If two dilutions of a sample give satisfactory
results &the variation in test results is less than 5%, then the average
value represents the BOD value. If the variation in the result is more
than 5%, then only the dilution which gives a greater DO depletion
is considered more significant and is recorded for use.
 Seeding

Seeding is inoculating organic waste matter with

live microorganisms, that can initiate
biodegradation(of organic matter) under
favorable conditions
 APPARATUS REQUIRED
BOD Incubator
Burette & Burette stand
300 mL glass stopper BOD bottles
500 mL conical flask
Pipettes with elongated tips
Pipette bulb
250 mL graduated cylinders
Wash bottle
 CHEMICALS REQUIRED
CHEMICALS—(ALL OF BEST QUALITY—ANALAR GRADE):-
Calcium Chloride Potassium iodide
Magnesium Sulphate Sodium azide
Ferric Chloride Concentrated sulfuric acid
Di Potassium Hydrogen Phosphate Starch indicator
Potassium Di Hydrogen Phosphate Sodium thiosulphate
Di sodium hydrogen phosphate Distilled or deionized
Ammonium Chloride
Manganous sulphate
Potassium hydroxide
Procedure
1. Dilution water- prepare dilution water by taking distilled
water in a bottle(about one litre per dilution per sample) and
aerate it using a compressor or magnetic stirrer or a fish tank
aerator for at least half an hour.(the duration of aeration is
governed by the requirement, that immediate DO should be
quite high –at least 7mg/l).Keep the bottle loosely plugged
with cotton, in a BOD incubator for at least 10 hours.(This
will hold DO in dilution water in equilibrium with the air in
the incubator at 20˚C- the temperature of the test)
Procedure
(2) Take about 300ml to400ml of dilution water in a1000ml jar& add
the nutrients phosphate buffer solution ,MgSO4 ,CaCl2,& FeCl3- 1ml
each.
(3) Shake the sample well and add a calculated quantity as per the
dilution adopted. Fill up the jar with dilution water to make up
1000ml.
(4) Mix the jar contents vertically without aerating .For this a long
glass tube(such as an old 50ml burette open at both ends without
the regulating cock) may be used
1. Fill up three BOD bottles A,B,C with the diluted sample. Tap the
bottles to release all entrapped air bubbles. Stopper the bottles.
Procedure
(6) Keep the bottles B & C in the incubator maintained at 20˚C.
(7) Find out immediate DO of bottle A(This should be done with in
15 minutes of diluting sample- to minimize errors due to the effects
of temperature gradients)
(8) After 5 days, determine DO of incubated samples in bottles B&C.
(9) Calculate BOD of the sample.
 CALCULATION
BOD5 at 20˚C =
{Immediate DO of diluted sample,(A)-DO5 of diluted & incubated
sample,(B/C)} x D.F.
CALCULATION
 Recommended standards
Maximum Remark
permissible BOD5 at s
20˚C
(1)Inland surface waters for use as raw 3mg/l BIS
water for public water supply& for bathing 2296

(2)Domestic sewage effluent discharged in 20mg/l BIS

to inland surface waters 4764

(3)Industrial effluents discharged in to 30mg/l BIS

inland surface waters 2490

(4)Treated effluents to be discharged on 200mg/l BIS

land 2490
(5)Industrial effluents falling into marine 100mg/l BIS
coastal areas 7968