FACULTY : ENGINEERING

TECHNOLOGY
LABORATORY: CIVIL
ENGINEERING TECHNOLOGY
EXPERIMENT: BIOCHEMICAL
OXYGEN DEMAND (BOD)

EDITION:
REVISION NO:
EFFECTIVE DATE:

2
FEB. 2015

AMENDMENT DATE:

FACULTY OF ENGINEERING TECHNOLOGY

DEPARTMENT OF CIVIL ENGINEERING TECHNOLOGY

ENVIRONMENTAL ENGINEERING TECHNOLOGY

LABORATORY
LABORATORY INSTRUCTION SHEETS
COURSE CODE

BNP 20503

EXPERIMENT CODE

EXPERIMENT 4

EXPERIMENT TITLE

BIOCHEMICAL OXYGEN DEMAND (BOD)

DATE
GROUP NO.
LECTURER/ INSTRUCTOR/
TUTOR
DATE OF REPORT
SUBMISSION

DISTRIBUTION OF MARKS
FOR LABORATORY REPORT:

EXAMINER COMMENTS:

1)
2)

ATTENDANCE/PARTICIPATION/DISPLINE

/5%

INTRODUCTION:

/5%

PROCEDURE:

/5%

RESULTS & CALCULATIONS

ANALYSIS

/15%
/15%

DISCUSSIONS:
ADDITIONAL QUESTIONS:

/20%
/15%

CONCLUSION:

/10%

SUGGESTION & RECOMENDATIONS

/5%

REFERENCES:

/5%

TOTAL:

/100%

RECEIVED DATE AND STAMP:

FACULTY : ENGINEERING
TECHNOLOGY
LABORATORY: CIVIL
ENGINEERING TECHNOLOGY
EXPERIMENT: BIOCHEMICAL
OXYGEN DEMAND (BOD)

EDITION:
2

REVISION NO:
EFFECTIVE DATE:

FEB. 2015

AMENDMENT DATE:

STUDENT CODE OF ETHICS

DEPARTMENT OF CIVIL ENGINEERING TECHNOLOGY
FACULTY OF ENGINEERING TECHNOLOGY

I hereby declare that I have prepared this report with my own efforts. I also
admit to not accept or provide any assistance in preparing this report and
anything that is in it is true.
1) Group Leader
Name
:
Matrix No. :

__________________________________________(Signature)
__________________________________
__________________________________

2) Group Member 1
Name
:
Matrix No :

__________________________________________(Signature)
__________________________________
___________________________________

3) Group Member 2
Name
:
Matrix No. :

__________________________________________(Signature)
__________________________________
__________________________________

4) Group Member 2
Name
:
Matrix No. :

__________________________________________(Signature)
__________________________________
__________________________________

FACULTY : ENGINEERING
TECHNOLOGY
LABORATORY: CIVIL
ENGINEERING TECHNOLOGY
EXPERIMENT: BIOCHEMICAL
OXYGEN DEMAND (BOD)

EDITION:
2

REVISION NO:
EFFECTIVE DATE:

FEB. 2015

AMENDMENT DATE:

1.0 OBJECTIVES
a) To understand the concept of biochemical oxygen demand in a given
water/wastewater sample.
b) To determine the biochemical oxygen demand in water/wastewater
sample.
2.0 LEARNING OUTCOMES
At the end of this course students are able to:
a) Describe the importance of BOD in the environmental studies.
b) Evaluate the water parameter quality in domestic wastewater sample and
water supply.
c) Identify problems and use their generic skill to solve the problems.
d) Develop their ability to work in group.
3.0 INTRODUCTION
3.1 Biochemical Oxygen Demand (BOD) test measures the ability of naturally
occurring microorganism to digest organic matter, usually in 5 days
incubation at 20oC by analysing the depletion of oxygen. BOD is the most
commonly used parameter for determining the oxygen demand on the
receiving water of a municipal or industrial discharge. BOD can also be
used to evaluate the efficiency of treatment processes and is an indirect
measure of biodegradable organic compounds in water.
3.2 The BOD test is normally required by a regulatory program. For this
experiment two analytical methods for testing and reporting BOD 5 will be
examined; dilution method (EPA method 405.1 and Standard method
5210 B) and respirometric method (using BODTrak apparatus by
Hachcompany).

FACULTY : ENGINEERING
TECHNOLOGY
LABORATORY: CIVIL
ENGINEERING TECHNOLOGY
EXPERIMENT: BIOCHEMICAL
OXYGEN DEMAND (BOD)

EDITION:
2

REVISION NO:
EFFECTIVE DATE:

FEB. 2015

AMENDMENT DATE:

4.0 INSTRUMENTS /APPARATUS / CHEMICAL / REAGENTS

4.1

Chemicals / Reagents
a)

pH calibration buffers: pH 4.00, pH 7.00 and pH 10.00

b)

Nutrient salt solutions (prepared by technician);

i)

Phosphate buffer solution: 8.5 g potassium dihydrogen phosphate

(KH2PO4) are dissolved in approximately 500 mL distilled water and
21.75 g Di potassium hydrogen phosphate (K2HPO4) are added,
followed by 33.4 g di sodium hydrogen phosphate (Na 2HPO4. 7H2O)
and 1.7 g ammonium chloride (NH4Cl). The solution is made up to
1 L and the pH of this buffer should be 7.2.

ii)

Magnesium sulphate solution: 22.5 g magnesium sulphate

(MgSO4. 7H2O) is dissolved in distilled water and the solution made
up to 1 L.

iii) Calcium chloride solution: 27.5 g of anhydrous calcium chloride

(CaCl2) are dissolved in distilled water and the solution made up to
1 L.
iv) Ferric chloride solution: 0.25 g FeCl3.6H2O is dissolved in distilled
water and the solution made up to 1 L.
c)

Dilution water: 1 mL of each of the nutrient salt solutions is added to

1 L deionised water and aeration carried out for several days (at least
12 hours) in the dark. Mix thoroughly and bring temperature to 20 +
3oC.
Note: Prepare dilution water 3 to 5 days before initiating BOD test to
ensure that the BOD of the dilution water is less than 0.2 mg/L.

d)

4.2

1 N of sulfuric acid, H2SO4 or 1 N sodium hydroxide, NaOH

Apparatus /Instruments
a)

300 mL glass stoppers BOD bottles

b)

Incubator regulated at 20+ 1oC

c)

pH meter

d)

Dissolved oxygen meter

e)

Air pump

f)

Measuring cylinder, 500 mL

2

FACULTY : ENGINEERING
TECHNOLOGY
LABORATORY: CIVIL
ENGINEERING TECHNOLOGY
EXPERIMENT: BIOCHEMICAL
OXYGEN DEMAND (BOD)

g)

Beaker

h)

Volumetric flask, 1000 mL

EDITION:
REVISION NO:
EFFECTIVE DATE:

2
FEB. 2015

AMENDMENT DATE:

5.0 PROCEDURE
5.1 Determination of sample size
Use formulas in Calculation section to calculate the minimum and
maximum estimated of sample size.
5.2 Pre-treatment of samples (Neutralizing samples)
Caustic alkalinity or acidity can prevent bacteria from growing during the
course of the BOD test. To prevent this, samples which have pH values
higher than pH 8.0 or lower than pH 6.0 must be neutralized to pH 7.0
before the test is performed.

(i).

Pour 50 mL of sample into a 100 mL beaker.

(ii).

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.

(iii).

Add 1 N sulfuric acid, H2SO4 if the sample is alkaline, or 1 N

sodium hydroxide, NaOH if the sample is acidic, until the pH
reaches 6.5-7.5.

(iv).

Calculate the amount of sulfuric acid or sodium hydroxide needed

to neutralize 1000 mL of the sample.

(v).

Add the calculated amount of acid or base to the sample (See

calculation section).

(vi).

Repeat steps 1-5 until the pH test shows pH 6.5-7.5. Record the
volume of NaOH or H2SO4 in Results section.

5.3 BOD5 measurement

(i)

Prepared the sample of estimated BOD5 (400 mg/L for influent and
60 mg/L for effluent).

FACULTY : ENGINEERING
TECHNOLOGY
LABORATORY: CIVIL
ENGINEERING TECHNOLOGY
EXPERIMENT: BIOCHEMICAL
OXYGEN DEMAND (BOD)

(ii)

EDITION:
REVISION NO:
EFFECTIVE DATE:

2
FEB. 2015

AMENDMENT DATE:

Label separately 6 unit of 300 mL BOD bottle. One bottle for blank
and another bottle for sample.

(iii)

For each bottle, partially filled with dilution water and add volume
of sample size based on your calculation (except the blank bottle).
Completely fill the bottles with dilution water.

(iv)

Determine the initial DO for each bottle and record data on

Results section.

(v)

Insert the bottle stoppers and will the top of each bottle each
round the bottle stopper with dilution water.

(vi)

Place the remaining bottles in the incubator at 20C and incubate

for five days.

(vii)

At the end of exactly five days (+3 hours), determine the final DO
content of the incubated bottles. Record the result on data sheet.

Note: 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. In any event, do not use the
depletion obtained as a blank correction.

FACULTY : ENGINEERING
TECHNOLOGY
LABORATORY: CIVIL
ENGINEERING TECHNOLOGY
EXPERIMENT: BIOCHEMICAL
OXYGEN DEMAND (BOD)

6.0

EDITION:
2

REVISION NO:
EFFECTIVE DATE:

FEB. 2015

AMENDMENT DATE:

RESULTS & CALCULATIONS

6.1

Results
a)

Type of sample: _________________________

b)

Initial pH of sample : _________________________

c)

Adjustment pH of sample : _________________________

d)

Volume of acid used

e)

Volume of base used : _________________________ mL

f)

Volume of sample used:

: _________________________ mL
(i). __________________ ml
(ii). _________________ ml
(iii). ---------------------- ml

Table 6.1: BOD5 of water sample

Sample

Volume of

Volume of

Initial

Final

DO

incubation

sample

DO,

DO,

depletion,

bottle, mL

size, mL

mg/L

mg/L

mg/L

BOD5,
mg/L

Blank
1
2
3
4
5
6.1.1
g)

Effluent or wastewater sample:

Type of sample: _________________________

h) Initial pH of sample

: _________________________

i)

Adjustment pH of sample : _________________________

j)

Volume of acid used

: _________________________ mL

k)

Volume of base used

: _________________________ mL

FACULTY : ENGINEERING
TECHNOLOGY
LABORATORY: CIVIL
ENGINEERING TECHNOLOGY
EXPERIMENT: BIOCHEMICAL
OXYGEN DEMAND (BOD)

EDITION:
2

REVISION NO:
EFFECTIVE DATE:

FEB. 2015

AMENDMENT DATE:

Table 6.1.2: BOD5 concentrated of effluent water or wastewater

samples

Sample

Volume of

Volume of

Initial

Final

DO

incubation

sample

DO,

DO,

depletion,

bottle, mL

size, mL

mg/L

mg/L

mg/L

BOD5,
mg/L

Blank
1
2
3
4
5

6.2 Calculations
6.2.1

Suppose the estimated BOD of an influent sample is 400 mg/L

and assume the DO of saturated dilution water is 8.0 mg/L. since
the criteria for most valid results states that the DO depletion at
the end of five days incubation should be at least 2.0 mg/L and
the residual DO at least 1.0 mg/L , the formulas to calculate the
minimum and maximum estimated dilution are as follows .

(i)

mL sample added to BOD bottle,

= ( minimum allowable depletion, mg/L x Volume of BOD bottle, mL )
estimated BOD , mg/L.

example:
minimum mL sample = [( 8 mg/L 6 mg/L ) x 300 mL ] / 400 mg/L
minimum mL sample = ( 2 x 300 ) / 400 = 600 / 400 = 1.5 mL

FACULTY : ENGINEERING
TECHNOLOGY
LABORATORY: CIVIL
ENGINEERING TECHNOLOGY
EXPERIMENT: BIOCHEMICAL
OXYGEN DEMAND (BOD)

EDITION:
2

REVISION NO:
EFFECTIVE DATE:

FEB. 2015

AMENDMENT DATE:

(ii) ml sample added to BOD bottle

= ( maximum allowed depletion , mg/l x Volume of BOD bottle, ml )

estimated BOD, mg/l.
example:
maximum mL sample = [ ( 8 mg/L 1 mg/L ) x 300 mL ] / 400 mg/L
maximum mL sample = ( 7x 300 ) / 400 = 5.24 mL

6.2.2 Calculate the amount of 1 N sodium hydroxide, NaOH or 1 N sulfuric

acid, H2SO4 needed to neutralize the sample to pH 7.0 using the
following formula:
Volume of acid or base needed, mL
= (volume acid or base used, mL x total test sample, mL)
Sample portion used for neutralization, mL.
For example, suppose 1.3 mL of 1 N NaOH are used to neutralize 50 mL of
sample to pH 7.0. Calculate the volume of NaOH to be added to neutralize
the sample as follows:
Volume 1 N NaOH needed, mL = (1.3 mL x 1000 mL) = 26 mL
50 L
6.2.3 To determine the value of the BOD5 in mg/L, use the following formula;
BOD5, mg/L = (DO depletion) x 300
Sample size, mL
7.0 ANALYSIS
Please analyze the data and results obtained in this experiment

FACULTY : ENGINEERING
TECHNOLOGY
LABORATORY: CIVIL
ENGINEERING TECHNOLOGY
EXPERIMENT: BIOCHEMICAL
OXYGEN DEMAND (BOD)

8.0

EDITION:
2

REVISION NO:
EFFECTIVE DATE:

FEB. 2015

AMENDMENT DATE:

DISCUSSIONS
Discuss your results both on the basis of any theory presented and on their
relevance to practical applications and current industrial practise. Comment
on the variation of your results and compare them with the recommended
standard values from the Department of Environment, Ministry of Natural
Resources and Environment or other international organization such as EPA,
WHO, EC, etc.

9.0

ADVANCED QUESTIONS

1. Why is it important to determine BOD value in wastewater treatment?

2. Why do you incubate BOD bottles?
3. Why you need to determine the BOD of the blank (dilution water, unseeded)?
4. List the chemical added to the dilution water and explain why they are
added?
5.

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?

6.

CONCLUSION
Conclusion is merely a summary, presented in a logical order, of the
important findings already reported in the discussion section. It also relates
to the objectives stated earlier.

Prepared by/Disediakan oleh :

Approved by/Disahkan oleh :

Signature/Tandatangan :
Name/Nama : DR. NOR HASLINA HASHIM
Date/Tarikh : FEBRUARY 2015

Signature/Tandatangan :
Name/Nama : DR. SURAYA HANI ADNAN
Date/Tarikh : FEBRUARY 2015