EXPERIMENT TITTLE :

Biochemical Oxygen Demand (BOD)

1.0 OBJECTIVE
Measured the strength of the water sample based on the amount of oxygen needed to stabilize
the organic matter in the sample.
2.0 THEORY
Biological Oxygen Demand (BOD) is a measure of the oxygen used by
microorganisms to decompose this waste. If there is a large quantity of organic waste in the
water supply, there will also be a lot of bacteria present working to decompose this waste. In
this case, the demand for oxygen will be high (due to all the bacteria) so the BOD level will
be high. As the waste is consumed or dispersed through the water, BOD levels will begin to
decline.
Dissolved oxygen (DO) is the amount of oxygen that is dissolved in water and is
essential to healthy streams and lakes. 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 and much of the available dissolved oxygen is
consumed by aerobic bacteria, robbing other aquatic organisms of the oxygen they need to
live. The dissolved oxygen level can be an indication of how polluted the water is and how
well the water can support aquatic plant and animal life. Generally, a higher dissolved oxygen
level indicates better water quality. When BOD levels are high, dissolved oxygen (DO) levels
decrease because the oxygen that is available in the water is being consumed by the bacteria.
Since less dissolved oxygen is available in the water, fish and other aquatic organisms may
not be able to survive.

3.0 PROCEDURE BOD

1. Pre-treatment of sample

Test pH sample (use pH meter)

pH neutral (6.5 8.5)

pH acidic (0 6.5) + alkali (NaOH)

pH alkali (8.5 14) + acid (HCl)

Use 100 mL of sample

2. Label separately 3 units of 300ml BOD bottles.

3. Prepare the sample of estimated BOD5 is 60 - 80mg/L.
4. Determine the sample size (7.5 26 ml)
5. Two bottles for water sample (replicate)

For each BOD bottles, partially filled with dilution water, carefully measure
out the proper volume of sample size. Add dilution water until the bottles are
completely filled.

One for blank (dilution water)

6. Determine the initial DO for each bottle.

7. Insert the bottle stopper.
8. Place the bottles in the incubator at 20C and incubate for _______days.
9. At the end of exactly ________ day (+/- 3 hours), test the final DO of the incubated
bottles.
- increase water sample temperature from 20C to room temperature before test the
final DO
10. Calculate the BOD for each dilution.
11. The most accurate BOD will be obtained from those dilutions that have a depletion of
at least 1.0 mg/l DO residual.
12. The dilution water blanks are used only to check the quality of the dilution water. If
the Quality of the water is good and free from impurities, the depletion of DO should
be less than 0.2 mg/l

4.0 RESULT AND DATA ANALYSIS

RESULT
Sample
type

Sample
ID

Volume
Sample
( mL)

BOD5

Blank
1

0
120

0
0.40

10.11
7.78

Final
DO
(mg/L
)
8.20
7.03

120

0.40

7.85

5.02

Dilution
Factor

Initial
DO
(mg/L)

DO
Depletion
(mg/L)

BOD
(mg/L)

BOD
Average

1.91
0.75

0
1.88

4.48

2.83

7.08

DATA ANALYSIS
Dilution Factor

volume of water

Volume of water plus dilution water

= 120 mL / 300 mL
= 0.40
BOD1 =

DOi - DOf
P

( 10.11 mg/L 8.20 mg/L )

0.40

=
BOD2 =

1.88 mg/L
( 7.78 mg/L 7.03 mg/L )
0.40

=
BODaverage

7.08 mg/L
= = BOD1+BOD2
2
= ( 1.88 mg/L + 7.08 mg/L )
2
= 4.48 mg/L

Sampel
pH

: 7.29

20 C

Source :

Drain

Time (day)

BODt

1
5

15.6
83.4

Time

BODt

1/ 3

0.400
0.391

Pretreatment: Alkalinity/Acidity
Sample volume : 15 mL
In NaOH
In N2SO4

: 0.005 mL
:

- mL

BOD Data Table

Date/Time Started : 13/1/2011, 9.00 am
Temperature

: 25 C
Graph (Time/BODt)1/3 versus Time (t)

Dilution water blank

From the result, the DOi blank water =10.11mg/L
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DOf blank water =8.20 mg/L

Thus, the deplete is 0.09 mg/L which is less than 2.0mg/L.
It had met the criteria.

Time

BODt

Graph

1/ 3

vs time

Rate constant, K =2.61

B

A

Where A = interception of graph=0.346

B = the slope of graph= (0.390-0.346)/ 5

K= 2.61

0.0088

0.346

(from the graph)

=0.066 day-1

5.0 QUESTION
Q1.
a) Calculate the min / max volume of sample to be added to prepare as estimated BOD
_____________ mg/L.
Min value: 120 ml
Max value: 420 ml
Minimum mL sample added to BOD bottle:
2 mLx 300 mL
=120 mL
5 mL
Maximum mL sample added to BOD bottle :
7 mLx300 mL
=420 mL
5 mL

b) Why must samples containing cautic alkalinity or acidity be adjusted before preparing
BOD dilution?
Caustic alkalinity and acidity can prevent the growth of bacteria during the test which
prevents the use of oxygen
c) The completed composite sample arrive in the lab at eg: (Tuesday, 18th January 2011)
What is the latest day and time the sample can be started for BOD.
d) Describe the function of BOD bottles cap and seal water.
The function of BOD bottles cap is to prevent evaporation of the water seal during the fiveday incubation period. It is also to prevent the air bubble in the bottle.

Q2.
a) Why must samples containing residual chlorine be dechlorinated before preparation of
BOD dilutions?
There are bacteria in a sample and we need to dechlorinated it to prohibit the growth of the
bacteria during the BOD test and also to prevents the use of oxygen
b) What reagents are required to chemically dechlorinate a BOD sample?
Sodium sulfite solution, Potassium iodide solution, acetic acid or sulfuric acid

c) What must be done to samples which have been dechlorinated or adjusted for pH
variations?
The samples will be seeded using the seed correction that we have obtained from the
calculation.
Q3.
a) State the formula to calculate
seed correction
Seed correction=

Seed BOD x mL seed per bottle

300

If <2 mg/ L D. O consumed TOO WEAK

If <1 mg/L D. O consumed TOO STRONG

BOD5 (seeded)
BOD 5 ( seeded )
( Initial D. Oi Final D .Of seed correction ) x 300 mL samplebottle

If <2 mg/ L D. O consumed TOO WEAK

If <1 mg/L D. O consumed TOO STRONG

b) Calculate the seed correction and BOD5 (seeded) for the data given as below
BOD5 of Seed Material
Dilution
#1

95 mg/L

mL of seed material

2 mL

mL of sample
Start D.O.

100 mL
7.8 mg/L
7

Final D.O.
Seed correction=

2.9 mg/L

2 mL x 95 mg/L
300

190
300

0.63 mg/ L
BOD 5=

(7.8 mg/ L 2.9 mg/L 0.63) x 300

100

12.81mg/ 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?
Volume=30 ml
DOi=7.3 mg/ L

DOf =1.8 mg/ L

P=30 mL /300 mL=0.1

BOD=

(7.3 1.8)
0.1

55 mg/L

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, ii) BOD3 and iii) BOD remaining at 3
days
i.

BODu=BO D5
8

(1 ekt )
0.23 (5 )

250/(1 e

365.84 mg/ L

ii.

BO D3=366(1 e0.23 (5 ))
250.11 mg/ L

iii.

BOD remaining at 3 days

BODu BO D3
365.84 250.11

115.73 mg/ L

Q5.
a) What is seeding process in BOD measurement?
Seeding process is a process that added the dilution water to the waste sample. It is a way to
add the population of microorganisms in the sample.
b) Explain preparation of seed material.
The material that have been selected will be seeded and will be placed in the suitable
container and incubate 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.
c) What materials can be used to seed a BOD sample
We can use any material which is can provide a suitable population of organisms.

Q6.
What is the significance of dissolved oxygen ?
The titrimetric iodometric methods will be conducted to measure the DO. There will be
significance of dissolved oxygen if a reaction occurred.

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.
Duplicate refers to making one copy while replicate refers to making copies of no specified
number, but they're effectively synonymous.
b) When are DO levels at their highest and their lowest and why?
Photosynthesis occurs during the day and it adds DO, hence the level of DO is highest at this
time. It will drops when at night because of the respiration of organisms living.
When BOD levels are high, dissolved oxygen (DO) levels decrease because the oxygen that
is available in the water is being consumed by the bacteria and it becomes less dissolved
oxygen is available in the water aquatic organisms may not survive.

DISCUSSION
The pH of sample must around 6.8 to 7.3 and the temperature around 20C. The
value of DO must more than 9mg/L. It is to ensure bacteria in sample are alive and
active. This experiment been carried out in 5 days. Reagent added into sample as a food
of bacteria (Phosphate Buffer, Magnesium Sulfate, Calcium Chloride, Ferric Chloride).
As seen in BOD table and data analysis, values of DO are decreased.
When BOD levels are increased, DO levels are decreased because the demand for
oxygen by the bacteria is high and it consumes oxygen that been dissolved from water

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sample. The factors influence a value of BOD is contamination, supersaturating

temperature, pressure, source water, and measurement of DO. The BOD 1 and BOD2
have average BOD for 4.48.
The sample is actually acidic and after pretreatment with alkaline NaOH 0.0005
ml and this microbial utilization possibly occurs oxidation and reduction process
simultaneously. The action of bacteria or microorganisms can cause color, taste, odor
problems. With some circumstances, anaerobic or unstable end product could happen so
more oxygen will oxidized rapidly to be a stable end product. This situation will
severely affect the ecology of the system.
In recommendation, one should not mix biodegradable material in water system
because biodegradable material usually consist of starches, fats, proteins, alcohols,
acids and esters which can be dissolved as food to microorganisms within a reasonable
time. This is also to prevent affecting ecology as mentioned above.

CONCLUSION
The objective of this experiment is successful which is to measure the strength of
the water sample based on the amount of oxygen needed to stabilize the organic matter
in the sample is achieved. The BOD test takes 5 days to complete and is performed
using a dissolved oxygen test kit. The BOD 5 value is 4.48 mg/L. From the table, it
shows that this water quality is from surface water type and contains few pollutants and
organic waste. When BOD levels are high, there is a decline in DO levels. This is
because the demand for oxygen by the bacteria is high and they are taking that oxygen
from the oxygen dissolved in the water. If there is no organic waste present in the water,
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there won't be as many bacteria present to decompose it and thus the BOD tend to be
lower and the DO level will be higher. From the result, we know that the drainage at
UTHM is poor and polluted and have to treat to collect clean water for various
purposes.

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