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Centre for Diploma Studies Group 2 (Section 1)
Department of Civil Engineering Session 2015/2016
itle! "ioc#emical $%&gen Demand E%periment Semester 2
1.0 Introduction
Microorganisms such as bacteria are responsible for decomposing organic waste. When
organic matter such as dead plants, leaves, grass clippings, manure, sewage, or even food
waste is present in a water supply, the bacteria will begin the process of breaking down this
waste. When this happens, much of the available dissolved oxygen is consumed by aerobic
bacteria, robbing other aquatic organisms of the oxygen they need to live. Biochemical
oxygen demand or BO is a procedure to determine the amount of oxygen consumed by the
microorganisms in the waste water chemically. !t is commonly used as the indicator to show
the cleanliness of the waste water.
2.0 Objective
"o measures the strength of the water sample #water, wastewater, etc.$ based on the
amount of oxygen needed to stabili%e the organic matter in the sample.
• "o assess the quality of surface waters.
• "o understand the characteristics of O contained in water and wastewater.
2.0 Equipment and Material/Reagent
1. 5 units o !00 ml "O# bottles
2. Incubator$ capable o maintaining 20 % 1&'
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Centre for Diploma Studies Group 2 (Section 1)
Department of Civil Engineering Session 2015/2016
itle! "ioc#emical $%&gen Demand E%periment Semester 2
!. 100 ml bea(er
). 100 ml graduated c*linder
5. 2 units o 25 ml meaning pipettes
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Centre for Diploma Studies Group 2 (Section 1)
Department of Civil Engineering Session 2015/2016
itle! "ioc#emical $%&gen Demand E%periment Semester 2
+. #O meter
,. p- meter
. ospate "uer
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Centre for Diploma Studies Group 2 (Section 1)
Department of Civil Engineering Session 2015/2016
itle! "ioc#emical $%&gen Demand E%periment Semester 2
issolve &.' g of KH 2 PO
4 , ().* g of K 2 HPO
4 , ++. g of Na2 HPO
4 and ).* g
NH 4Cl of in deioni%ed water. -dust p/ to *.(, if necessary, with either ) 0
H 2
SO4 or 0aO/. ilute to one liter.
. Magnesium ulpate
issolve ((.' g Mg SO4.7 H
2O and dilute to one liter.
10. 'alcium 'loride
issolve (*.' g CaCl2 and dilute to one liter.
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Centre for Diploma Studies Group 2 (Section 1)
Department of Civil Engineering Session 2015/2016
itle! "ioc#emical $%&gen Demand E%periment Semester 2
11. 3erric 'loride
issolve 1.(' g Fe Cl3.6
H 2O and dilute to one liter.
0O"23 "o prepare dilution water, add ) ml of each of the four solutions listed above to
one liter of deioni%ed water. 4aturate with O by drawing a vacuum through the solution.
).0 Metods and rocedure
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Centre for Diploma Studies Group 2 (Section 1)
Department of Civil Engineering Session 2015/2016
itle! "ioc#emical $%&gen Demand E%periment Semester 2
). '11 ml of waste water is collected from "asik 5+ using a beaker.
(. "he sample water is then tested by using p/ meter. !t is tested to be neutral. "he readingsof both p/ and temperature are recorded in the Bench 4heet.
/owever, if the sample is not neutral, pre6treatment process has to be done to neutrali%e
the sample. 7rocedure for neutrali%ing samples3
a$ 7our '1 m8 of samples into a )11 m8 beaker. b$ Measure the p/ of the sample using a p/ meter. !f the p/ is out of the range of
p/ 9.1 to p/ &.1 continue with steps + : 9, otherwise perform the BO test on the
untreated sample.
c$ -dd ) 0 sulfuric acid if the sample is alkaline, or )0 sodium hydroxide if the
sample is acidic, until the p/ reaches *.1.d$ ;alculate the amount of sulfuric acid or sodium hydroxide needed to neutrali%e
)111ml of the sample.e$ -dd the calculated amount of acid or base to the sample.
f$ <epeat steps )6' until the p/ test shows p/ *.1.
+. ( ml of sample water is poured into the first, second and the third bottle. "hen, all the
bottle if filled with dilution water until +11ml.Only the last bottle is filled with dilution
water and labelled as =Blank )>.
. - magnetic stirrer is then added into these two bottles respectively. "he sample in the
bottle is stirred whilst the O #issolved Oxygen$ meter is put into the bottle to get the
O reading. <epeat this step for the second and third bottle.
'. "he readings of O for all three bottles are recorded in the Bench 4heet as initial O,
9. "he bottles are kept in the BO incubator and remained around (1 ℃ .
*. "he final O is recorded after ' days of the experiment.
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Centre for Diploma Studies Group 2 (Section 1)
Department of Civil Engineering Session 2015/2016
itle! "ioc#emical $%&gen Demand E%periment Semester 2
5.0 Results and 4nal*sis
"'$CE'C*+ $,-GE. DE*.D ( "$D)
Date! 2//2016
ime ! 110 am
Analyst:
Sample Details:
Source! G lae
p 0C
61 250
Pretreatment:
*lalinit&/*cidit&
Sample 3olume ! 500 m+
' . .ao ! 0 m+' . .2S$4 ! 0 m+
Comments!
.eutral #is is ecause t#e sample ater #as a p of 61
#ic# is considered as neutral (65 7 5) 8 neutral
Sample&pe
Sample'D
3olumeSample( m+)
Dilution9actor
'nitial D$(mg/+)
9inal D$( mg/+)
D$Depletion( mg/+)
"$D(mg/+)
"$D1
"lan 2 ml 0006 :1 66 025 1
"$D2
"lan 2 ml 0006 : 4 06 5
"$D
"lan 2 ml 0006 ;1 2 05; ;65
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Centre for Diploma Studies Group 2 (Section 1)
Department of Civil Engineering Session 2015/2016
itle! "ioc#emical $%&gen Demand E%periment Semester 2
Average BOD( s#o t#e calculation)
Cancelled Data/ <esult! *verage 8 1 = 5 = ;65 8 5:20 mg/+
"$D1 8 1 mg/+
"$D2 8 5 mg/+
"$D 8 ;65 mg/+
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Centre for Diploma Studies Group 2 (Section 1)
Department of Civil Engineering Session 2015/2016
itle! "ioc#emical $%&gen Demand E%periment Semester 2
+.0 #44 46478I
+.1 'alculate or te average "O#9
-verage ?37.31+53.73+86.57
3 ? '@.(1 mgA
+.2 o: all te calculation and state i an* o te data needs to tro: out.
O epletion # mgA8$ ? !nitial O : inal O
O) ? *.@) : *.99
? 1.(' mgA8
O( ? *.*@ : *.+
? 1.+9 mgA8
O+ ? *.&) : *.(+
? 1.'& mgA8
BO' ? !nitial O : inal O
7
BO) ? 1.('A1.119*
? +*.+) mgA8
BO( ? 1.+9A1.119*
? '+.*+ mgA8
BO+ ? 1.'&A1.119*
? &9.'* mgA8
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Centre for Diploma Studies Group 2 (Section 1)
Department of Civil Engineering Session 2015/2016
itle! "ioc#emical $%&gen Demand E%periment Semester 2
+.! e dilution :ater blan( cannot deplete more tan 0.2 mg/7. ;as tis criteria met9
"he BO blank did not show a O depletion of more than 1.( mgA8 after the five day incubation
period. - drop of more than 1.( mgA8 indicates some type of contamination or calibration error.4o the criteria is met.
+.) #oes *ou sample so:s <a to=ic eect>9
"he sample is not showing any toxic effect, it is because the waste water, took from a lake is not
a toxic source. -nd the p/ value of the sample is nearly neutral. !t means that the sample doesn>t
show any toxic effect.
+.5 'ould *ou rel* on *our "O# results9 ;*9
Ces, because the average BO result is compared with the effluent standard #O2, Malaysia$
+.+ "* reerring to e:age/Eluent tandard ?#OE$ Mala*sia@ could *our sample be
discarge to river untreated9 I not$ suggest te associated treatment or "O# removal.
-ccording to the 4ewageA2ffluent 4tandard #O2,Malaysia$, the sample cannot be discharge to
the river untreated because the value obtained with ( ml of wastewater is higher than '1 mgA8
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Centre for Diploma Studies Group 2 (Section 1)
Department of Civil Engineering Session 2015/2016
itle! "ioc#emical $%&gen Demand E%periment Semester 2
,.0 #iscussion
D). a@ 'alculate te min / ma= volume o sample to be added to prepare as estimated
"O# ?mg/7@
2xample3
-ssume that the estimated BO of an influent sample is 11mgA8.
"he .O. of saturated dilution water is approximately &.1mgA8.
.O. depletion to be at least (.1mgA8 after ' days of incubation and the residual .O. to be
at least ).1mgA8.
4ample Water, m8
? (min
/max allowabledepletion , mg
/ L) x(
vol of BOD bottle, mL) Etimated BOD(mg/ L)
4ample water minimum, m8
?2mg/ L x 300mL
400mg/ L
? ).' m8
Max allowable depletion
? #4at .O. concentration$ : #Min residual concentration$
? #&.1 : ).1$ mgA8
? *.1 mgA8
4ample water maximum, m8
?7 mg/ L x300mL
400mg/ L
? '.(' m8
b@ ;* must samples containing cautic al(alinit* or acidit* be adjusted beore
preparing dilution9
"he BO dilution must be natural which range between p/ values of 9.' to *.' because
microbes are most active in this range of p/ value.
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Centre for Diploma Studies Group 2 (Section 1)
Department of Civil Engineering Session 2015/2016
itle! "ioc#emical $%&gen Demand E%periment Semester 2
c@ e completed composite sample arrive in te lab at 11.!0 4.M $;ednesda*$ 2
Marc 201+.;at is te latest da* and time te sample can be started or "O#.
"he sample must be prepared not more than ( hours. !f it is begin within ( hours of
collection, cold storage is unnecessary but, if the test will begin within more than ( hours,
the sample must keep at or below ℃ during compositing
d@ #escribe te unction o "O# bottle>s cap and seal :ater.
"he function of BO bottle>s cap and seal water is to avoid drawing air into the dilution
bottle and reduce evaporation of the water seal during incubation. !t was used to ensure
that no air bubble is trapped in the bottle that will affect the characteristics of sample. !t is
also to prevent further oxygen dissolving in. Otherwise, bottle>s cap was using to reduce
evaporation of the water seal during evaporation.
D(. a@ ;* must samples containing residual clorine be declorinated beore preparation
o "O# dilutions9
"o determine the amount of valuable bacteria from its sources. ;hlorine can kill the
microbes live and bacteria in samples.
b@ ;at reagents are required to cemicall* declorinate a "O# sample9
4ulphuric acid solution or acetic acid solution, potassium iodide solution, sodium sulphite
#0a(4O+$
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Centre for Diploma Studies Group 2 (Section 1)
Department of Civil Engineering Session 2015/2016
itle! "ioc#emical $%&gen Demand E%periment Semester 2
c@ ;at must be done to samples :ic ave been declorinated or adjusted or p-
variations9
"he samples which have been dechlorinated must be seeded with +11m8 of BO water in
the BO bottle before allows it store in the BO incubator.
D+. a$ tate te ormula to calculate
i seed correction
4eed correction ?BOD
5of eed mate!ial x vol"me of ample
300
ii@ "O#5 ?seeded@
BOD5 #mg A8$ ?
[ ( D1− D
2 )−( B1−B
2) ] P
WhereE
D1 ? O of diluted sample immediately after preparation, mgA8
D2 ? O of diluted sample after ' days incubation at (1 ;, mgA8 ⁰
7 ? decimal volumetric fraction of sample used
b$ ;alculate the seed correction and BO' #seeded$ for the data given as below
"O#5 o eed Material 5 mg/7ilution
F)
m8 of seed material ( m8
m8 of sample )11 m8
4tart .O. *.& mgA8
inal .O. (.@ mgA8
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Centre for Diploma Studies Group 2 (Section 1)
Department of Civil Engineering Session 2015/2016
itle! "ioc#emical $%&gen Demand E%periment Semester 2
olution3
4eed correction ?BOD5 of eed mate!ial x vol"me of ample
300
?95 x 2
300
? 1.9+++ mgA8
BOD5
#mgA 8$ ?
D1− D
2
P 7 ?
2
100
? 0.02
? (7.8 # 2.9)
0.02
? (' mgA8
D.
a@ !0 m7 o :aste:ater are placed in a !00 m7 "O# bottle. e sample is diluted to ill te
bottle. e #O concentrations at te beginning and te end o 5Ada* incubation period are
,.! mg/7 and 1. mg/7 respectivel*. ;at is te "O#9
olutionB
7 ?30mL
300mL
? 1.)
BOD5 #mg A8$ ?
D1− D
2
P =
7.3−1.8
0.1=55mg/ L
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Centre for Diploma Studies Group 2 (Section 1)
Department of Civil Engineering Session 2015/2016
itle! "ioc#emical $%&gen Demand E%periment Semester 2
b@ e "O#5 o a :aste:ater :as determined to be 250 mg/7. I te reaction coeicient
:as 0.2! l/d$ calculate
i$ ultimate BO,
L0=
$ m
1−e−%t
¿
250
1−e−0.23( 5)
¿ +9' mg/ L
ii$ BOD3 and
BODt = L0(1−e
−%t )
BOD3=365 (1−e
−(0.23) (3) )
¿181.9mg/ L
iii$ BO remaining at + days
BO' GBOD
3 ? BOD8 # BO at & days$
BOD8=365 (1−e
−(0.23) (8 ))
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Centre for Diploma Studies Group 2 (Section 1)
Department of Civil Engineering Session 2015/2016
itle! "ioc#emical $%&gen Demand E%periment Semester 2
¿301mg / L
C5. a@ ;at is seeding process in "O# measurement9
4eeding is a process of adding live bacteria and microorganism to a sample. !f the
samples tested contain materials which could kill or inure the microorganisms, the
condition must be corrected and healthy active organisms added.
b@ E=plain preparation o seed material.
• 4elect a material to be used for seeding which will have a BO of at least )&1
mgA8. "his will help ensure that the seed correction meets the 1.9 mgA8 minimum
specified in H4tandard MethodsI.
• 7lace the material in a suitable container and incubate at (1J; for (hours.
Ksually, settled raw domestic sewage prepared in the manner above will have
sufficient BO for use as a seed material. !f not, small quantities of digester
supernatant, return activated sludge, or an acclimated seed material can be used to
increase the potency of the seed material used for the test. -s an alternative,
commercially available seed material may be used. "he seed correction should not
exceed ).1 mgA8 BOE therefore, care should be taken not to use too strong a seed
material for the test.
• "he key to a good seed correction is a relatively stable seed material which
produces a good seed correction in every test situation
c@ ;at materials can be used to seed a "O# sample
-ny source of water which can provide a suitable population of organisms can be
used, however, settled raw sewage or commercially prepared seed material are the
most common sources
C+. ;at is te signiicance o dissolved o=*gen 9
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Centre for Diploma Studies Group 2 (Section 1)
Department of Civil Engineering Session 2015/2016
itle! "ioc#emical $%&gen Demand E%periment Semester 2
issolved oxygen allows animals to breathe in water and it provides a suitable habitat
for the other animals. Bacteria in the water also use this oxygen to break down animals
and plants. "he oxygen level is not very high in most water sources. !f the oxygen
level is reduced, the animals begin to die. !t is also to maintaining the aquatic life and
aesthetic quality of streams and lakes.
C,. a@ ;it regard to precision$ ten percent duplicate or replicate samples sould be
run. is :ould result in one duplicate sample or one replicate sample being run
ever* ten samples. #ierentiate replicate and duplicate sample
• <eplicates are two or more separate water samples collected in the field from the
same site and depth. <eplicates are used to determine the errors involved in sample
collectionE if there are no errors in the collection and analysis, and then the difference
between two replicate analyses indicates the natural variability in the water at that
location.
• uplicates are two Lor more lab analyses on the same water sample. uplicates are
used to determine the N difference between two samples in order to estimate the error
involved in the analyses.
b@ ;en are #O levels at teir igest and teir lo:est and :*9
O levels are the highest if the sample of water is not polluted means there is no toxic
and bacterial effect ."herefore, the sample of water has a lot of oxygen demanding
wastes that well within self6purification capacity. Otherwise, the BO level will be
low which indicates the water sample does not have any sign of live microbes. "hen,
O levels are low if the sample water does not have enough oxygen to dissolve in
water that is needed by all living things.
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Centre for Diploma Studies Group 2 (Section 1)
Department of Civil Engineering Session 2015/2016
itle! "ioc#emical $%&gen Demand E%periment Semester 2
.0 'onclusion
"he result shows that in + beaker of BO, the highest O is BO beaker +. "his show that the
higher the O depletion the higher the BO. !n this + beaker of sample has the same amount of
volume sample which is (m8. "he initial of the O between the beakers is different. O
depletion that we get is final O minus with initial O. "his show that the higher the O
depletion the higher the BO. BO in beaker ) has the smallest O depletion and beaker ( was
in between beaker ) and beaker +. -fter that, we find the average of the result that we get. !n
beaker ), we got +*.+)mgA8. !n beaker (, we got '+.*+mgA8. While in beaker +, we got
&9.'*mgA8. "hen we plus this + result and divide by + to get the average result and we get
'@.(1mgA8. in this + beaker of sample has the same amount of volume sample which is (m8.
"his show that the amount of oxygen needed to stabili%e the organic matter in the sample based
on the measurement of strength of the water sample.
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Centre for Diploma Studies Group 2 (Section 1)
Department of Civil Engineering Session 2015/2016
itle! "ioc#emical $%&gen Demand E%periment Semester 2
.0 4ppendi=
7icture ) shows four BO bottles, which for the BO bottle ), ( and + are the mixture of water
samples and dilution water while BO bottle is only dilution water.
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Centre for Diploma Studies Group 2 (Section 1)
Department of Civil Engineering Session 2015/2016
itle! "ioc#emical $%&gen Demand E%periment Semester 2
7icture ( shows one of the group members was measured the p/ value of the sample.
7icture + shows all the BO bottles were placed in the incubator for 5 da&s #ic# capaleof maintaining 20 > 1 0c
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Centre for Diploma Studies Group 2 (Section 1)
Department of Civil Engineering Session 2015/2016
itle! "ioc#emical $%&gen Demand E%periment Semester 2
7icture ' shows our group members measured the 7/ value of each sample in BO bottles after ' days placed in an incubator.
10.0 Reerence
)$ avis - ;ornwell, Macken%ie 8 avis. !ntroduction to 2nvironment 2ngineering. +rd 2dition.
Mc5raw /ill, (1)1.
($ issolved Oxygen #Wikipedia$.
http3AAen.wikipedia.orgAwikiAissolvedPoxygen #-ccessed (1)96()6+$
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