Laboratory Module

EXPERIMENT 2

BIOCHEMICAL OXYGEN DEMAND (BOD)

OBJECTIVES

To understand the nature of oxygen demand in water by determining

the degree of water pollution caused by microorganisms through
biodegradation.

To determine the biochemical oxygen demand (BOD) of the given

wastewater samples.

2.0 INTRODUCTION

BIOCHEMICAL OXYGEN DEMAND (BOD)

BOD is the amount of oxygen required by bacteria to stabilize organic

matter under aerobic conditions. The BOD test involves the
determination of oxygen uptake by bacteria under standard conditions
(5 day incubation at 20 C). Strong wastewater samples must be
diluted in keeping with the solubility of oxygen to ensure that DO will
be present throughout the test period.

The following guide may be used for preparing dilutions:

Typical Amount of Resulting Source

 mg/L
Concentratio
sample
Sample BOD5 range n
to add
(%)
(mL)
Clear creek water 0 to 5 300 100
Clear creek water 4 to 10 150 50
Clear creek water 8 to 20 75 25
Weak sewage, polluted creek water 10 to 25 60 20
Weak sewage, polluted creek water 13 to 33 45 15
Weak sewage, polluted creek water 20 to 50 30 10
Weak sewage, polluted creek water 25 to 62.5 24 8
Weak sewage, polluted creek water 40 to 100 15 5
Strong sewage, industrial waste 50 to 125 12 4
Strong sewage, industrial waste 67 to 167 9 3
Strong sewage, industrial waste 150 to 250 6 2
Strong sewage, industrial waste 200 to 500 3 1
Strong sewage, industrial waste 400 to 1000 1.5 0.5
Strong sewage, industrial waste 667 to 1667 0.9 0.3
2000 to
Strong sewage, industrial waste 0.3 0.1
5000
4000 to
Strong sewage, industrial waste 0.15 0.05
10000

3.0 MATERIALS AND APPARATUS

3.1 Reagents

a) Phosphate buffer solution

b) Magnesium sulphate solution
c) Calcium chloride solution
d) Ferric chloride solution

3.2 Apparatus

a) BOD sample bottle with stopper

b) 50 mL pipettes
c) 1 mL pipettes
d) 10 L water container
e) DO meter

5.0 RESULTS
Record your sample details. Fill in your DO data to the table below.

Table 1: DO concentration

Dissolved oxygen (DO)

Bottle *
(mg/L)
Day 0
Blank
Day 5
Day 0
Seed control
Day 5

Sample dilution

Day 0
100% sample concentration
Day 5
Day 0
50% sample concentration
Day 5
Day 0
25% sample concentration
Day 5
Day 0
20% sample concentration
Day 5
Calculate the BOD for the samples.

When dilution water is seeded,

BOD (mg/l) = [(D1 D2) (B1 B2) x f] P

(1)

When dilution water is not seeded,

BOD (mg/l) = [(D1 D2)] P

(2)

where,

D1 = DO of diluted sample immediately after

preparation, mg/l
D2 = DO of diluted sample after 5-day incubation,
mg/l
B1 = DO of seed control before incubation, mg/l
B2 = DO of seed control after incubation, mg/l
f = fraction of seeded dilution water volume in
sample to volume of seeded dilution water in
seed control (volume of seed in sample/volume
of seed control)
 P = fraction of wastewater sample volume to total
combined volume (volume of sample/volume of
mixture)

BOD can be calculated using the formula given below:

BOD (mg/L) = [ (D1 D2) (B1 B2) x f ] / P

Table 2: BOD calculation

Blan Seed Sample dilution

k Ctrl 100% 50% 25% 20%
(A) Sample vol. in vol. of bottle
(mL)
(B) Seed vol. in vol. of bottle (mL)
(C) Initial DO, Day 0 (mg/L)
(D) Final DO, Day 5 (mg/L)
(E) DO depletion (mg/L), (C) - (D)
(F) f, ((E) x [(B) / vol. of bottle]
(G) P, (A) / vol. of bottle
(H) BOD5 (mg/L), [(E) - (F)] / (G)

6.0 DISCUSSION

Briefly discuss your experiments and results.

7.0 CONCLUSION

Conclude your experimental results.

8.0 REFERENCES

8.1 Dunnivat, F.M. (2004). Environmental Laboratory Exercise for Instrumental

Analysis and Environmental Chemistry, Wiley, Interscience.
8.2 APHA., AWWA., & WPCF. (1992). Standard Methods for Examination of Water and
Wastewater. 16th Edition. American Public Health Association, Washington.

9.0 QUESTIONS

9.1 What is the need for using seeded dilution water in the BOD test?

9.2 What are the main sources of error in a BOD test?

9.3 Why is a nitrification inhibitor used in the test?

9.4 What is the significance of BOD?

9.5 A 20 mL wastewater sample is placed in a standard 300 mL BOD

bottle and the bottle is filled with dilution water. The bottle had an
initial dissolved oxygen concentration of 10 mg/L and a final
concentration of 3 mg/L. A blank (a BOD bottle filled with dilution
water) run in parallel showed no changes in dissolved oxygen over
the 5 day incubation period. Calculate the BOD5.

9.6 In a test for determination of BOD of a sample of water from a

polluted stretch of river, three dilutions were used. The
observations are given below. Calculate the BOD 5 of the sample.
Then, from the result discuss what is the discharge limit for BOD
according to Environmental Quality Act (EQA) ?

Sample Dilution, % Initial DO, Final DO,

No mixture mg/L mg/L
1 50 7.5 2.5
2 20 7.5 5.2
3 10 7.7 6.9