EXPERIMENT ON THE DETERMINATION OF BIOCHEMICAL OXYGEN DEMAND (BOD)

BOD
The biochemical oxygen demand determination 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.
BOD of water or polluted water is the amount of oxygen, required for the biological
decomposition of dissolved organic matter to occur under standard condition at a
standardized time and temperature. Usually, the time is taken as 5 days and the temperature
is 20˚C.
The test measures the molecular oxygen utilized during a specified incubation period for the
biochemical degradation of organic material (carbonaceous demand) and the oxygen used to
oxidize inorganic material such as sulphides and ferrous ion. It also may measure the amount
of oxygen used to oxidize reduced forms of nitrogen (nitrogenous demand).
Apparatus/Equipment Reagents
 BOD Incubator  Calcium chloride  Potassium iodide
 Burette & Burette  Magnesium sulphate  Sodium azide
Stand  Ferric chloride  Concentrated
 300 mL glass BOD  Dipotassium hydrogen sulphuric acid
stopper BOD bottles phosphate  Starch indicator
 500 mL conical flask  Potassium dihydrogen  Sodium thiosulfate
 Pipettes with phosphate  Distilled or deionized
elongated tips  Disodium hydrogen water
 Pipette bulb phosphate
 250 mL graduated  Ammonium chloride
cylinders  Manganous sulphate
 Wash bottle  Potassium hydroxide
Procedure
 Dissolve manganese sulphate in 1L of  For the starch indicator, weigh
distilled water and filter the solution. approximately 2g of starch and
Take 364g of the solution and place it dissolve it in 100ml of hot distilled
in the beaker. Place it in magnetic water. To preserve thesolution, add
stirrer to dissolve the content. 0.2g of salicylic acid.
 Take 700 g of potassium hydroxide  Weigh 22.5 g of magnesium sulphate
and add 150 potassium iodide and and dissolve it in distilled water. Take
dissolve it in a distilled water to make 100ml standard measuring flask and
1l of the solution. Dissolve 10g of place funnel over it. Transfer it in to
sodium azide in 40ml of distilled water the 100 ml standard flask and make up
and add this with constant stirring to to 100ml using distilled water.
cool the alkaline iodide solution  Weigh 0.15g of ferric chloride and
prepared. dissolve it in distilled water. Take
 Weigh approximately 25g of sodium 100ml standard flask and make up to
thiosulphate and dissolve it in distilled 100ml using distilled water.
water and make a 1l solution. Add 1 g
of sodium hydroxide to preserve it.
  Weigh 8.5 of potassium di hydrogen
phosphate and dissolve it in distilled
water then add exactly 21.75g of di
potassium hydrogen phosphate and
dissolve it. To the same beaker, add
33.4g of di sodium hydrogen
phosphate is added. Finally, add 1.7g
of ammonium chloride and dissolve it.
Take 1l measuring flask and add
distilled water until the solution
becomes 1l.
 Take 5l of organic free distilled water
and add 5ml of calcium chloride,
magnesium sulphate solution and
ferric chloride solution and the
phosphate buffer solution. Prepare
dilution of water 3 to 5 days before
initiating BOD test to ensure that the
BODof the dilution water is less than
0.2mg/L
TESTING OF SAMPLE
 Take four 300 mL glass stoppered BOD
bottles (two for the samples and two
for the blank)
 Add 10 mL of the sample to each of the
two BOD bottles and fill the remaining
quantity with the dilution water.
 The remaining two BOD bottles are for
blank, to these bottles add dilution
water alone.
 After the addition immediately place
the glass stopper over the BOD bottles
and note down the numbers of the
bottle for identification.
 Now preserve one blank solution bottle
and one sample solution bottle in a
BOD incubator at 20˚C for five days.
 The other two bottles (one blank and
one sample) needs to be analysed
immediately.
 Add 2 mL of manganese sulphate to
the BOD bottle by inserting the
calibrated pipetted just below the
surface of the surface of the liquid.
 Add 2 mL of alkali-iodize-azide reagent
in the same manner.
 If oxygen present, a brownish-orange
cloud of precipitate of floe will appear.
 Add 2 mL of alkali-iodized-azide
reagent in the same manner.
 Allow it to settle for sufficient time in
order to react completely with oxygen.
 When this floc has settled to the
bottom, shake the contents thoroughly
by turning it upside down.
 Add 2 mL of concentrated sulphuric
acid via a pipette held just above the
surface of the sample.
 Carefully stopper and invert several
times to dissolve the floc.
 Titration needs to be started
immediately after the transfer of the
contents to Erlenmeyer flask.
 Rinse the burette with sodium
thiosulfate and then fill it with sodium
thiosulfate. Fix the burette to the
stand.
 Measure out 203 mL of the solution
from the bottle and transfer to an
Erlenmeyer flask.
 Titrate the solution with standard
sodium thiosulfate solution until the
yellow colour of liberated iodine is
almost faded out.
 Add 1 mL of starch solution and
continue the titration until the blue
colour disappears to colourless.
 Note down the volume of the sodium
thiosulfate solution added, which give
the D.O. in mg/L. Repeat the titration
for concordant values.
 After 5 days, take out the bottles from
the BOD incubator and analyse the
sample and the blank for DO.
 Add 2 mL of manganese sulphate to
the BOD by inserting the calibrated
pipette just below the surface of the
liquid.
 Allow it to settle for sufficient time in
order to react completely with oxygen.
 When this floc has settled to the
bottom, shake the contents thoroughly
by turning it upside down.
  Add 2 mL of concentrated sulphuric
acid via a pipette held just above the
surface of the sample.
 Carefully stopper and invert several
times to dissolve the floc.
 Titration needs to be started
immediately after the transfer of the
contents to Erlenmeyer flask.
 Rinse the burette with sodium
thiosulfate and then fill it with
thiosulfate. Fix the burette to stand.
 Measure out 203 mL of the solution
from the bottle and transfer to an
Erlenmeyer flask.
Calculation

BOD = initial DO – final DO