Centre for Diploma Studies
Group
2 (Section 1)
Department of Civil Engineering
Session
2015/2016
Title: Biochemical Oxygen Demand Experiment
Semester
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 BOD is a procedure to determine the amount of oxygen consumed by the
microorganisms in the waste water chemically. It is commonly used as the indicator to show
the cleanliness of the waste water.
2.0 Objective
To measures the strength of the water sample (water, wastewater, etc.) based on the
amount of oxygen needed to stabilize the organic matter in the sample.
To assess the quality of surface waters.
To understand the characteristics of DO contained in water and wastewater.
2.0 Equipment and Material/Reagent
1.
5 units of 300 ml BOD bottles
2. Incubator, capable of maintaining 20 1C
BOD EXPERIMENT |ENVIRONMENTAL ENGINEERING LAB REPORT
Centre for Diploma Studies
Group
2 (Section 1)
Department of Civil Engineering
Session
2015/2016
Title: Biochemical Oxygen Demand Experiment
Semester
3. 100 ml beaker
4. 100 ml graduated cylinder
5. 2 units of 25 ml meaning pipettes
BOD EXPERIMENT |ENVIRONMENTAL ENGINEERING LAB REPORT
Centre for Diploma Studies
Group
2 (Section 1)
Department of Civil Engineering
Session
2015/2016
Title: Biochemical Oxygen Demand Experiment
Semester
6. DO meter
7. pH meter
8. Phosphate Buffer
BOD EXPERIMENT |ENVIRONMENTAL ENGINEERING LAB REPORT
Centre for Diploma Studies
Group
2 (Section 1)
Department of Civil Engineering
Session
2015/2016
Title: Biochemical Oxygen Demand Experiment
Semester
Dissolve 8.5 g of KH 2 PO 4 , 21.7 g of K 2 HPO 4 , 33.4 g of
Na 2 HPO4 and 1.7 g
NH 4 Cl of in deionized water. Adjust pH to 7.2, if necessary, with either 1 N
H 2 SO 4 or NaOH. Dilute to one liter.
9. Magnesium Sulphate
Dissolve 22.5 g Mg SO 4 .7 H 2 O and dilute to one liter.
10. Calcium Chloride
Dissolve 27.5 g CaCl 2 and dilute to one liter.
BOD EXPERIMENT |ENVIRONMENTAL ENGINEERING LAB REPORT
Centre for Diploma Studies
Group
2 (Section 1)
Department of Civil Engineering
Session
2015/2016
Title: Biochemical Oxygen Demand Experiment
Semester
11. Ferric Chloride
Dissolve 0.25 g
Fe Cl3 .6 H 2 O and dilute to one liter.
NOTE: To prepare dilution water, add 1 ml of each of the four solutions listed above to
one liter of deionized water. Saturate with DO by drawing a vacuum through the solution.
4.0 Methods and Procedure
BOD EXPERIMENT |ENVIRONMENTAL ENGINEERING LAB REPORT
Centre for Diploma Studies
Group
2 (Section 1)
Department of Civil Engineering
Session
2015/2016
Title: Biochemical Oxygen Demand Experiment
Semester
1. 500 ml of waste water is collected from Tasik G3 using a beaker.
2. The sample water is then tested by using pH meter. It is tested to be neutral. The readings
of both pH and temperature are recorded in the Bench Sheet.
However, if the sample is not neutral, pre-treatment process has to be done to neutralize
the sample. Procedure for neutralizing samples:
a) Pour 50 mL of samples into a 100 mL beaker.
b) Measure the pH of the sample using a pH meter. If the pH is out of the range of
pH 6.0 to pH 8.0 continue with steps 3 6, otherwise perform the BOD test on the
untreated sample.
c) Add 1 N sulfuric acid if the sample is alkaline, or 1N sodium hydroxide if the
sample is acidic, until the pH reaches 7.0.
d) Calculate the amount of sulfuric acid or sodium hydroxide needed to neutralize
1000ml of the sample.
e) Add the calculated amount of acid or base to the sample.
f) Repeat steps 1-5 until the pH test shows pH 7.0.
3. 2 ml of sample water is poured into the first, second and the third bottle. Then, all the
bottle if filled with dilution water until [Link] the last bottle is filled with dilution
water and labelled as Blank 1.
4. A magnetic stirrer is then added into these two bottles respectively. The sample in the
bottle is stirred whilst the DO (Dissolved Oxygen) meter is put into the bottle to get the
DO reading. Repeat this step for the second and third bottle.
5. The readings of DO for all three bottles are recorded in the Bench Sheet as initial DO,
6. The bottles are kept in the BOD incubator and remained around 20 .
7. The final DO is recorded after 5 days of the experiment.
BOD EXPERIMENT |ENVIRONMENTAL ENGINEERING LAB REPORT
Centre for Diploma Studies
Group
2 (Section 1)
Department of Civil Engineering
Session
2015/2016
Title: Biochemical Oxygen Demand Experiment
Semester
5.0 Results and Analysis
BIOCHEMICAL OXYGEN DEMAND ( BOD)
Analyst:
Date: 2/3/2016
Time : 11.30 a.m
Sample Details:
Source: G3 lake
pH
6.71
25.0
Pretreatment:
Alkalinity/Acidity
Sample Volume : 500 mL
I N NaoH : 0 mL
I N N2SO4 : 0 mL
Comments:
Neutral. This is because the sample water has a pH of 6.71
which is considered as neutral. (6.5 7.5) = neutral
Sample
Type
Sample
ID
Volume
Sample
( mL)
Dilution
Factor
Initial DO
(mg/L)
Final DO
( mg/L)
DO
Depletion
( mg/L)
BOD
(mg/L)
Blank
2 ml
0.0067
7.91
7.66
0.25
37.31
Blank
2 ml
0.0067
7.79
7.43
0.36
53.73
Blank
2 ml
0.0067
7.81
7.23
0.58
86.57
BOD1
BOD2
BOD3
BOD EXPERIMENT |ENVIRONMENTAL ENGINEERING LAB REPORT
Centre for Diploma Studies
Group
2 (Section 1)
Department of Civil Engineering
Session
2015/2016
Title: Biochemical Oxygen Demand Experiment
Semester
Average BOD
( show the calculation)
Cancelled Data/ Result: Average = 37.31 + 53.73 + 86.57 = 59.20 mg/L
3
BOD1 = 37.31 mg/L
BOD2 = 53.73 mg/L
BOD3
= 86.57 mg/L
BOD EXPERIMENT |ENVIRONMENTAL ENGINEERING LAB REPORT
6.0
Centre for Diploma Studies
Group
2 (Section 1)
Department of Civil Engineering
Session
2015/2016
Title: Biochemical Oxygen Demand Experiment
Semester
DATA ANALYSIS
6.1 Calculate for the average BOD?
Average =
37.31+53.73+ 86.57
= 59.20 mg/
3
6.2 Show all the calculation and state if any of the data needs to throw out.
DO Depletion ( mg/L) = Initial DO Final DO
DO1
= 7.91 7.66
= 0.25 mg/L
DO2
= 7.79 7.43
= 0.36 mg/L
DO3
= 7.81 7.23
= 0.58 mg/L
BOD5 = Initial DO Final DO
P
BOD1 = 0.25/0.0067
= 37.31 mg/L
BOD2 = 0.36/0.0067
= 53.73 mg/L
BOD3 = 0.58/0.0067
= 86.57 mg/L
BOD EXPERIMENT |ENVIRONMENTAL ENGINEERING LAB REPORT
Centre for Diploma Studies
Group
2 (Section 1)
Department of Civil Engineering
Session
2015/2016
Title: Biochemical Oxygen Demand Experiment
Semester
6.3 The dilution water blank cannot deplete more than 0.2 mg/L. Was this criteria met?
The BOD blank did not show a DO depletion of more than 0.2 mg/L after the five day incubation
period. A drop of more than 0.2 mg/L indicates some type of contamination or calibration error.
So the criteria is met.
6.4 Does you sample shows a toxic effect?
The sample is not showing any toxic effect, it is because the waste water, took from a lake is not
a toxic source. And the pH value of the sample is nearly neutral. It means that the sample doesnt
show any toxic effect.
6.5 Could you rely on your BOD results? Why?
Yes, because the average BOD result is compared with the effluent standard (DOE, Malaysia)
6.6 By referring to Sewage/Effluent Standard (DOE, Malaysia) could your sample be
discharge to river untreated? If not, suggest the associated treatment for BOD removal.
According to the Sewage/Effluent Standard (DOE,Malaysia), the sample cannot be discharge to
the river untreated because the value obtained with 2 ml of wastewater is higher than 50 mg/L
BOD EXPERIMENT |ENVIRONMENTAL ENGINEERING LAB REPORT
10
Centre for Diploma Studies
Group
2 (Section 1)
Department of Civil Engineering
Session
2015/2016
Title: Biochemical Oxygen Demand Experiment
Semester
7.0 Discussion
Q1. a) Calculate the min / max volume of sample to be added to prepare as estimated
BOD (mg/L)
Example:
Assume that the estimated BOD of an influent sample is 400mg/L.
The D.O. of saturated dilution water is approximately 8.0mg/L.
D.O. depletion to be at least 2.0mg/L after 5 days of incubation and the residual D.O. to be
at least 1.0mg/L.
Sample Water, mL
=
(min/max allowable depletion, mg/ L) x (vol of BOD bottle , mL)
Estimated BOD(mg/ L)
Sample water minimum, mL
2 mg/ L x 300 mL
400 mg/L
= 1.5 mL
Max allowable depletion
= (Sat D.O. concentration) (Min residual concentration)
= (8.0 1.0) mg/L
= 7.0 mg/L
Sample water maximum, mL
=
7 mg/L x 300 mL
400 mg/ L
= 5.25 mL
b) Why must samples containing cautic alkalinity or acidity be adjusted before
preparing dilution?
The BOD dilution must be natural which range between pH values of 6.5 to 7.5 because
microbes are most active in this range of pH value.
BOD EXPERIMENT |ENVIRONMENTAL ENGINEERING LAB REPORT
11
Centre for Diploma Studies
Group
2 (Section 1)
Department of Civil Engineering
Session
2015/2016
Title: Biochemical Oxygen Demand Experiment
Semester
c) The completed composite sample arrive in the lab at 11.30 A.M ,Wednesday, 2
March [Link] is the latest day and time the sample can be started for BOD.
The sample must be prepared not more than 24 hours. If it is begin within 2 hours of
collection, cold storage is unnecessary but, if the test will begin within more than 2 hours,
the sample must keep at or below 4 during compositing
d) Describe the function of BOD bottles cap and seal water.
The function of BOD bottles cap and seal water is to avoid drawing air into the dilution
bottle and reduce evaporation of the water seal during incubation. It was used to ensure
that no air bubble is trapped in the bottle that will affect the characteristics of sample. It is
also to prevent further oxygen dissolving in. Otherwise, bottles cap was using to reduce
evaporation of the water seal during evaporation.
Q2. a) Why must samples containing residual chlorine be dechlorinated before preparation
of BOD dilutions?
To determine the amount of valuable bacteria from its sources. Chlorine can kill the
microbes live and bacteria in samples.
b) What reagents are required to chemically dechlorinate a BOD sample?
Sulphuric acid solution or acetic acid solution, potassium iodide solution, sodium sulphite
(Na2SO3)
BOD EXPERIMENT |ENVIRONMENTAL ENGINEERING LAB REPORT
12
Centre for Diploma Studies
Group
2 (Section 1)
Department of Civil Engineering
Session
2015/2016
Title: Biochemical Oxygen Demand Experiment
Semester
c) What must be done to samples which have been dechlorinated or adjusted for pH
variations?
The samples which have been dechlorinated must be seeded with 300mL of BOD water in
the BOD bottle before allows it store in the BOD incubator.
Q3. a) State the formula to calculate
i
seed correction
Seed correction =
ii)
BOD5 of seed material x volume of sample
300
BOD5 (seeded)
BOD 5 (mg /L) =
[ ( D 1D2 )( B1 B 2) ]
P
Where;
D 1 = DO of diluted sample immediately after preparation, mg/L
D 2 = DO of diluted sample after 5 days incubation at 20 C, mg/L
P = decimal volumetric fraction of sample used
b) Calculate the seed correction and BOD5 (seeded) for the data given as below
Dilution
BOD5 of Seed Material
mL of seed material
95 mg/L
2 mL
mL of sample
Start D.O.
Final D.O.
100 mL
7.8 mg/L
2.9 mg/L
#1
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Centre for Diploma Studies
Group
2 (Section 1)
Department of Civil Engineering
Session
2015/2016
Title: Biochemical Oxygen Demand Experiment
Semester
Solution:
Seed correction
BOD5 of seed material x volume of sample
300
95 x 2
300
= 0.6333 mg/L
BOD 5
(mg/ L)
D1 D2
P
P=
2
100
= 0.02
=
(7.8 2.9)
0.02
= 245 mg/L
Q4.
a) 30 mL of wastewater are placed in a 300 mL BOD bottle. The sample is diluted to fill the
bottle. The DO concentrations at the beginning and the end of 5-day incubation period are
7.3 mg/L and 1.8 mg/L respectively. What is the BOD?
Solution:
P=
30 mL
300 mL
= 0.1
BOD 5
(mg /L) =
D1 D2 7.31.8
=
=55 mg/L
P
0.1
BOD EXPERIMENT |ENVIRONMENTAL ENGINEERING LAB REPORT
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Centre for Diploma Studies
Group
2 (Section 1)
Department of Civil Engineering
Session
2015/2016
Title: Biochemical Oxygen Demand Experiment
Semester
b) The BOD5 of a wastewater was determined to be 250 mg/L. If the reaction coefficient
was 0.23 l/d, calculate
i) ultimate BOD,
L0 =
Ym
1ekt
250
0.23 ( 5)
1e
365 mg/ L
ii)
BOD 3 and
BOD t=L 0 ( 1ekt )
BOD 3=365 ( 1e( 0.23) (3) )
181.9 mg/L
iii) BOD remaining at 3 days
BOD5 + BOD 3 = BOD 8 ( BOD at 8 days)
BOD 8=365 ( 1e( 0.23) (8 ) )
BOD EXPERIMENT |ENVIRONMENTAL ENGINEERING LAB REPORT
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Centre for Diploma Studies
Group
2 (Section 1)
Department of Civil Engineering
Session
2015/2016
Title: Biochemical Oxygen Demand Experiment
Semester
301mg /L
Q5. a) What is seeding process in BOD measurement?
Seeding is a process of adding live bacteria and microorganism to a sample. If the
samples tested contain materials which could kill or injure the microorganisms, the
condition must be corrected and healthy active organisms added.
b) Explain preparation of seed material.
Select a material to be used for seeding which will have a BOD of at least 180
mg/L. This will help ensure that the seed correction meets the 0.6 mg/L minimum
specified in Standard Methods.
Place the material in a suitable container and incubate at 20C for 24hours.
Usually, settled raw domestic sewage prepared in the manner above will have
sufficient BOD for use as a seed material. If 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. As an alternative,
commercially available seed material may be used. The seed correction should not
exceed 1.0 mg/L BOD; therefore, care should be taken not to use too strong a seed
material for the test.
The key to a good seed correction is a relatively stable seed material which
produces a good seed correction in every test situation
c) What materials can be used to seed a BOD sample
Any 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
Q6. What is the significance of dissolved oxygen ?
BOD EXPERIMENT |ENVIRONMENTAL ENGINEERING LAB REPORT
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Centre for Diploma Studies
Group
2 (Section 1)
Department of Civil Engineering
Session
2015/2016
Title: Biochemical Oxygen Demand Experiment
Semester
Dissolved 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. The oxygen level is not very high in most water sources. If the oxygen
level is reduced, the animals begin to die. It is also to maintaining the aquatic life and
aesthetic quality of streams and lakes.
Q7. a) With regard to precision, ten percent duplicate or replicate samples should be
run. This would result in one duplicate sample or one replicate sample being run
every ten samples. Differentiate replicate and duplicate sample
Replicates are two or more separate water samples collected in the field from the
same site and depth. Replicates are used to determine the errors involved in sample
collection; 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.
Duplicates are two [or more] lab analyses on the same water sample. Duplicates are
used to determine the % difference between two samples in order to estimate the error
involved in the analyses.
b) When are DO levels at their highest and their lowest and why?
DO levels are the highest if the sample of water is not polluted means there is no toxic
and bacterial effect .Therefore, the sample of water has a lot of oxygen demanding
wastes that well within self-purification capacity. Otherwise, the BOD level will be
low which indicates the water sample does not have any sign of live microbes. Then,
DO levels are low if the sample water does not have enough oxygen to dissolve in
water that is needed by all living things.
BOD EXPERIMENT |ENVIRONMENTAL ENGINEERING LAB REPORT
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Centre for Diploma Studies
Group
2 (Section 1)
Department of Civil Engineering
Session
2015/2016
Title: Biochemical Oxygen Demand Experiment
Semester
8.0 Conclusion
The result shows that in 3 beaker of BOD, the highest DO is BOD beaker 3. This show that the
higher the DO depletion the higher the BOD. In this 3 beaker of sample has the same amount of
volume sample which is 2mL. The initial of the DO between the beakers is different. DO
depletion that we get is final DO minus with initial DO. This show that the higher the DO
depletion the higher the BOD. BOD in beaker 1 has the smallest DO depletion and beaker 2 was
in between beaker 1 and beaker 3. After that, we find the average of the result that we get. In
beaker 1, we got 37.31mg/L. In beaker 2, we got 53.73mg/L. While in beaker 3, we got
86.57mg/L. Then we plus this 3 result and divide by 3 to get the average result and we get
59.20mg/L. in this 3 beaker of sample has the same amount of volume sample which is 2mL.
This show that the amount of oxygen needed to stabilize the organic matter in the sample based
on the measurement of strength of the water sample.
BOD EXPERIMENT |ENVIRONMENTAL ENGINEERING LAB REPORT
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Centre for Diploma Studies
Group
2 (Section 1)
Department of Civil Engineering
Session
2015/2016
Title: Biochemical Oxygen Demand Experiment
Semester
9.0 Appendix
Picture 1 shows four BOD bottles, which for the BOD bottle 1, 2 and 3 are the mixture of water
samples and dilution water while BOD bottle 4 is only dilution water.
BOD EXPERIMENT |ENVIRONMENTAL ENGINEERING LAB REPORT
19
Centre for Diploma Studies
Group
2 (Section 1)
Department of Civil Engineering
Session
2015/2016
Title: Biochemical Oxygen Demand Experiment
Semester
Picture 2 shows one of the group members was measured the pH value of the sample.
Picture 3 & 4 shows all the BOD bottles were placed in the incubator for 5 days which capable
of maintaining 20 1 0c
BOD EXPERIMENT |ENVIRONMENTAL ENGINEERING LAB REPORT
20
Centre for Diploma Studies
Group
2 (Section 1)
Department of Civil Engineering
Session
2015/2016
Title: Biochemical Oxygen Demand Experiment
Semester
Picture 5 shows our group members measured the PH value of each sample in BOD bottles after
5 days placed in an incubator.
10.0 Reference
1) Davis A Cornwell, Mackenzie L Davis. Introduction to Environment Engineering. 3rd Edition.
McGraw Hill, 2010.
2) Dissolved Oxygen (Wikipedia).
[Link] (Accessed 2016-21-3)
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