1. PROCEDURE STEPS 1.

1 PRINCIPLE The BOD is the amount of oxygen required by bacteria for stabilization of decomposable organic matter under aerobic conditions. The quantity of oxygen required for the stabilization is taken as a measure of decomposable organic matter. The sample of waste or an appropriate dilution is incubated for 5 days at ! o" or # days at $o" in the BOD incubator. The reduction in DO concentration during the incubation period yields a measure of the biochemical oxygen demand. %erator BOD incubator %pparatus & BOD bottles '. Phosphate Buffer: Dissol(e ).5 g of *+ ,O-. '.$5 g * +,O-. ##.- g of /a +,O-0$+ O and '.$ g of /+-"l in one litre of distilled water. The p+ should be $. without further ad1ustment. . Ma !es"u# Su$phate: Dissol(e .5 g of 2g3O-.$+ O in little distilled water and dilute to one litre. #. %err"& Ch$or"'e So$ut"o!: Dissol(e !. 5 g of 4e"l#0 5+ O in little distilled water and dilute to one litre. -. Ca$&"u# Ch$or"'e So$ut"o!: Dissol(e $.5 g of "a"l in little distilled water and dilute to one litre. 5. See'"! : The purpose of seeding is to introduce into sample a biological population for oxidizing organic matter. 6here such micro7organisms are already present0 as in domestic water or biologically treated effluent0 it is not necessary to add the seed. 5. So'"u# Su$f"te so$ut"o! (.(2)N& Dissol(e '.5$5 g /a 3O#0 in distilled water and dilute to '!!!ml. $. G$u&ose* $uta#"& a&"' so$ut"o!& Dry reagent grade glucose and glutamic acid at '!#8" for ' hr. Dissol(e '5!mg glucose and '5!mg glutamic acid in distilled water and dilute to ' litre. %dditionally all reagents listed in DO estimation are required. '9 ,repare dilution water by aerating distilled water with supply of clean compressed air to get saturated with dissol(ed oxygen. 1

1.2 INSTRUMENT S 2. REAGENTS

+. PROCEDURE

%dd one ml each of phosphate buffer0 ferric chloride0 calcium chloride and magnesium sulphate solution to one litre of distilled water as the micro nutrients for bacteria. #9 3ample p+ should be ad1usted to $.!. 3eed the dilution water if required. -9 2ake se(eral dilution of sample to obtain required depletion. The following dilutions are suggested& !.' 7 '.! : for strong trade waste ' 7 5 : for raw and settled sewage 5 7 5 : for oxidized effluent 5 7 '!! : for polluted ri(er waters 59 ,repare the desired mixture by adding sample in dilution water. 4ill up one #!! ml bottle with the mixture and other one with dilution water ;blank9 in two sets. 59 *eep one set in BOD incubator for 5 days at ! 8" or # days at $o". 4ind out immediate DO of blank and sample bottle. $9 4ind out DO in both the bottles after incubation for 5 days. )9 To find the dissol(ed oxygen ;DO9 of sample and blank0 proceed as per the Dissol(ed Oxygen determination procedure. ,. CALCULATION '9 6hen dilution water is not seeded0 ;D'7D 9 x '!! BOD5 mg<litre = 77777777777777777777777 : of Dilution 9 6hen the dilution water is seeded0 ;D'7D 9 7;B'7B 9 x '!! BOD5 mg<litre = 7777777777777777777777777777777 : of Dilution 6here0 D' = DO of diluted sample immediately after preparation0 mg<l D = DO of diluted sample immediately after 5 days incubation period B' = DO of blank ; seeded diluting water9 before incubation 0mg<l B = DO of blank ; seeded diluting water9 after incubation 0mg<l 2