CENTRE FOR DIPLOMA STUDIES

DIPLOMA OF CHEMICAL ENGINEERING TECHNOLOGY

ENVIRONMENTAL ENGINEERING LABORATORY

LABORATORY INSTRUCTION SHEETS

COURSE CODE DAK 23503

EXPERIMENT CODE EXPERIMENT 4

EXPERIMENT TITLE BIOCHEMICAL OXYGEN DEMAND (BOD)

DATE
(INDIVIDUAL REPORT)

STUDENT NAME &

MATRIC NO. & SIGNATURE

INSTRUCTOR Ts. Dr. Hazlini Binti Dzinun

DATE OF REPORT SUBMISSION 1 WEEK AFTER EXPERIMENT DATE

Element Percentage
(%)
Attendance/Participation/Discipline /5
Procedure /10
Results & Calculation /10
Analysis /20
MARKS:
Discussion /20
Answer question /20
Conclusion /10
References /5
Total /100

RECEIVED DATE AND STAMP: EXAMINER COMMENTS:

 FACULTY: CENTRE FOR
EDITION: 1
DIPLOMA STUDIES
ENVIRONMENTAL ENGINEERING
REVISION NO: 0
LABORATORY DAK 23503
EXPERIMENT: EFFECTIVE DATE: 11/2/2016
BIOCHEMICAL OXYGEN DEMAND
(BOD) AMENDMENT DATE: 10/10/2019

1.0 OBJECTIVE OF EXPERIMENT

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.

2.0 LEARNING OUTCOMES

At the end of this course students are able to:

a) the student be able to describe the importance of BOD in the environmental studies.
b) the students be able to measure the BOD of samples with the right sample size.

3.0 INTRODUCTION / THEORY

3.1 Biochemical Oxygen Demand is a common, environmental procedure for determining the extent
to which oxygen within a sample can support microbial life. This method is popular in many
environmental laboratories analyzing waste water, compost, sludge, and soil samples. When
a measurement is made of all oxygen consuming materials in a sample, the result is termed
“Total Biochemical Oxygen Demand” (TBOD), or often just simply “Biochemical Oxygen
Demand” (BOD).

3.2 Because the test is performed over a five day period, it is often referred to as a “five Day BOD
“, or a BOD5. In addition, this procedure is only suitable for samples void of serious matrix
interferences. To gain a broader appreciation of oxygen demand, additional avenues of interest
may be explored including CBOD (carbonaceous oxygen demand), COD (chemical oxygen
demand), and TOC (total organic carbon).

3.3 Because of complications measuring this ultimate BOD (BODu), BODu is usually extrapolated
from laboratory 5-day BOD bottle tests BODt = BODu (1 – e-kt ). In many biological treatment
plants, the facility effluent large numbers of nitrifying organisms which are developed during
the treatment process. These organisms can exert an oxygen demand as they convert
nitrogenous compounds (ammonia and organic nitrogen) to more stable forms (nitrites and
nitrates). At least part of this oxygen demand is normally measured in a five day BOD.
Sometimes it is advantageous to measure just the oxygen demand exerted by organic
(carbonaceous) compounds, excluding the oxygen demand exeried by the nitrogenous
compounds. To accomplish this, the nitrifying organisms can be inhibited from using oxygen
by the addition of a nitrification inhibitor to the samples. The result is termed “Carbonaceous
Biochemical Oxygen Demand” or BOD.
 FACULTY: CENTRE FOR
EDITION: 1
DIPLOMA STUDIES
ENVIRONMENTAL ENGINEERING
REVISION NO: 0
LABORATORY DAK 23503
EXPERIMENT: EFFECTIVE DATE: 11/2/2016
BIOCHEMICAL OXYGEN DEMAND
(BOD) AMENDMENT DATE: 10/10/2019

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 (Na2HPO4. 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) 1 N of sulfuric acid, H2SO4 or 1 N sodium hydroxide, NaOH

4.2 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
g) Beaker
h) Volumetric flask, 1000 mL
 FACULTY: CENTRE FOR
EDITION: 1
DIPLOMA STUDIES
ENVIRONMENTAL ENGINEERING
REVISION NO: 0
LABORATORY DAK 23503
EXPERIMENT: EFFECTIVE DATE: 11/2/2016
BIOCHEMICAL OXYGEN DEMAND
(BOD) AMENDMENT DATE: 10/10/2019

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).
(ii) 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 20°C 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: CENTRE FOR
EDITION: 1
DIPLOMA STUDIES
ENVIRONMENTAL ENGINEERING
REVISION NO: 0
LABORATORY DAK 23503
EXPERIMENT: EFFECTIVE DATE: 11/2/2016
BIOCHEMICAL OXYGEN DEMAND
(BOD) AMENDMENT DATE: 10/10/2019

6.0 RESULTS & CALCULATIONS

6.1 Results

a) Type of sample:____________________________
b) Initial pH of sample:_________________________
c) Adjustment of pH sample:____________________
d) Volume of acid used (mL):____________________
e) Volume of base used (mL):____________________
f) Volume of sample used (mL): (i) ________________
(ii) ________________
(iii)________________

Table 6.1: BOD5 of blank sample

Sample Volume of Volume of Initial DO, Final DO BOD5,

incubation sample size, mg/L DO, depletion, mg/L
bottle, mL mL mg/L mg/L

Blank

6.1.1 Effluent water or wastewater sample (minimum sample):

g) Type of sample:____________________________
h) Initial pH of sample:_________________________
i) Adjustment of pH sample:____________________
j) Volume of acid used (mL):____________________
k) Volume of base used (mL):____________________

Table 6.1.1: BOD5 (minimum sample) of wastewater samples

Sample Volume of Volume of Initial DO, Final DO BOD5,

incubation sample size, mg/L DO, depletion, mg/L
bottle, mL mL mg/L mg/L

1
2
 FACULTY: CENTRE FOR
EDITION: 1
DIPLOMA STUDIES
ENVIRONMENTAL ENGINEERING
REVISION NO: 0
LABORATORY DAK 23503
EXPERIMENT: EFFECTIVE DATE: 11/2/2016
BIOCHEMICAL OXYGEN DEMAND
(BOD) AMENDMENT DATE: 10/10/2019

6.1.2 Effluent water or wastewater sample (maximum sample):

l) Type of sample:____________________________
m) Initial pH of sample:_________________________
n) Adjustment of pH sample:____________________
o) Volume of acid used (mL):____________________
p) Volume of base used (mL):____________________

Table 6.1.2: BOD5 (maximum sample) of wastewater samples

Sample Volume of Volume of Initial DO, Final DO BOD5,

incubation sample size, mg/L DO, depletion, mg/L
bottle, mL mL mg/L mg/L

1
2

6.2 Calculations

6.2.1 The two equations below to determine the maximum and minimum amount of sample water
to be used in dilutions.

Sample water, mL= (minimum allowable depletion, mg/L) x (volume of BOD bottle, mL)
Estimated BOD, mg/L

Sample water, mL= (maximum allowable depletion, mg/L) x (volume of BOD bottle, mL)
Estimated BOD, mg/L

7.0 ANALYSIS

Please analyze the data and results obtained in this experiment.

7.1 Calculate for the average BOD

7.2 Show all the calculation and state if any of the data needs to throw out.
7.3 The dillution water blank cannot deplete more than 0.2 mg/L. Was this criteria met?
7.4 Could you rely on your BOD results? Why?
7.5 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.
 FACULTY: CENTRE FOR
EDITION: 1
DIPLOMA STUDIES
ENVIRONMENTAL ENGINEERING
REVISION NO: 0
LABORATORY DAK 23503
EXPERIMENT: EFFECTIVE DATE: 11/2/2016
BIOCHEMICAL OXYGEN DEMAND
(BOD) AMENDMENT DATE: 10/10/2019

8.0 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?

10.0 CONCLUSION

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