CENTER FOR DIPLOMA STUDIES

DEPARTMENT OF CIVIL ENGINEERING

ENVIRONMENTAL ENGINEERING LABORATORY

LABORATORY INSTRUCTION SHEETS

COURSE CODE DAC 10903

COURSE NAME ENVIRONMENTAL ENGINEERING

EXPERIMENT TITLE
BIOCHEMICAL OXYGEN DEMAND
 STUDENT CODE OF ETHIC
(SCE)
DEPARTMENT OF CIVIL ENGINEERING
CENTRE FOR DIPLOMA STUDIES

We, hereby confess that we have prepared this report on our own effort. We also admit not to receive any help
from any third party during the preparation of this report and pledge that everything mentioned in the report is
true.

1) Group Leader __________________________________ (Signature)

Name : __________________________________
Matrix No. : __________________________________

2) Group Member 1 ___________________________________ (Signature)

Name : ___________________________________
Matrix No : ___________________________________

3) Group Member 2 __________________________________ (Signature)

Name : __________________________________
Matrix No. : __________________________________

4) Group Member 3 __________________________________ (Signature)

Name : __________________________________
Matrix No. : __________________________________
 Faculty: Centre for Diploma Studies (CeDS)

Department: Civil Engineering

Title: BIOCHEMICAL OXYGEN DEMAND ( BOD)

1.0 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.
_____________________________________________________________________________________

2.0 LEARNING OUTCOMES

i) the student be able to describe the importance of BOD in the environmental studies.
ii) the students be able to measure the BOD of samples with the right sample size.
______________________________________________________________________________
3.0 THEORY

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). 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).

Because of complications measuring this ultimate BOD (BOD u), BODu is usually extrapolated from
-kt
laboratory 5-day BOD bottle tests BODt = BODu ( 1 – e )

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 Diploma Studies (CeDS)
Department: Civil Engineering

Title: BIOCHEMICAL OXYGEN DEMAND ( BOD)

4.0 EQUIPMENT & REAGENTS :

1. 5 units 300 mL glass stopper BOD Bottles

0
2. Incubator , capable of maintaining 20 ± 1 C
3. 100 mL beaker
4. 500 mL measuring cylinder
5. 1000 mL Volumetric flusk
6. Dissolved Oxygen meter
7. pH meter
8. Phosphate Buffer
Dissolve 8.5g KH2PO4, 21.7g K2HPO4, 33.4g Na2HPO4, and 1.7g NH4Cl in deionized water. Adjust
pH to 7.2, if necessary, with either 1 N H2SO4 or NaOH. Dilute to one liter.

9. Magnesium Sulfate
Dissolve 22.5g MgSO4.7H2O and dilute to one liter.

10. Calsium Chloride

Dissolve 27.5g CaCl2 and dilute to one liter.

11. Ferric Choloride

Dissolve 0.25g FeCl3.6H2O and dilute to one liter.

NOTE: To prepare dilution water, add one 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.

5.0 PROCEDURES

1. Determination of sample size

Use formulas in Calculation section to calculate the minimum and maximum estimated of
sample size.

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.
 Faculty: Centre for Diploma Studies (CeDS)
Department: Civil Engineering
Title: BIOCHEMICAL OXYGEN DEMAND ( BOD)

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.

6.0 RESULTS AND CALCULATION

Write the results in benchcheet

7.0 DATA ANALYSIS

7.1 Calculate for the average BOD

7.2 Show all the calculation and state if any of the data needs to thrown out.

7.3 The dillution water blank can not deplete more than 0.2 mg/L. Was this criteria met?

7.4 Does you sample shows ’a toxic effect’?

7.5 Could you rely on your BOD results? Why?

7.6 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.

8.0 DISCUSSION

Q1. a) Calculate the min / max volume of sample to be added to prepare as estimated
BOD _____________ mg/L.

b) Why must samples containing cautic alkalinity or acidity be adjusted before preparing BOD
dilution?

c) The completed composite sample arrive in the lab at

eg:_________________,_______________) What is the latest day and time the sample can
be started for BOD.

d) Describe the function of BOD bottle’s cap and seal water.

 Faculty: Centre for Diploma Studies (CeDS)
Department: Civil Engineering
Title: BIOCHEMICAL OXYGEN DEMAND ( BOD)

Q2. a) Why must samples containing residual chlorine be dechlorinated before preparation of
BOD dilutions?

b) What reagents are required to chemically dechlorinate a BOD sample?

c) What must be done to samples which have been dechlorinated or adjusted for pH
variations?

Q3. a) State the formula to calculate

i) seed correction
ii) BOD5 (seeded)

b) Calculate the seed correction and BOD5 (seeded) for the data given as below
BOD5 of Seed Material 95 mg/L
Dilution #1 mL of seed material 2 mL
mL of sample 100 mL
Start D.O. 7.8 mg/L
Final D.O. 2.9 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?

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

Q5. a) What is seeding process in BOD measurement?

b) Explain preparation of seed material.
c) What materials can be used to seed a BOD sample

Q6. What is the significance of dissolved oxygen ?

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

b) When are DO levels at their highest and their lowest and why?
Bench Sheet

BIOCHEMICAL OXYGEN DEMAND ( BOD)

Analyst:
Date:

Time :

Sample Details:

Source:

0
pH C

Pretreatment:

Alkalinity/Acidity Comments:

Sample Volume : __________mL

I N NaoH : __________mL
I N N2SO4:__________mL

Volume
DO
Sample Sample Sample Dilution Initial DO Final DO BOD
Depletion
Type ID ( mL) Factor (mg/L) ( mg/L) (mg/L)
( mg/L)

Blank
BOD---

Blank
BOD---

Blank
BOD---

Average BOD
( show the calculation)
Cancelled Data/ Result:

BOD__ =

BOD__ =

BOD__ =