INTRODUCTION

Biochemical oxygen demand (BOD) is defined as the amount of oxygen

required by bacteria, while stabilizing decomposable organic matter under
aerobic conditions.

The significance of a BOD test is to examine the amount of oxygen required to

determine the pollution strength of domestic and industrial wastes & purity of
water bodies. It is one of the most important tests in regulating stream-
pollution-control activities. The BOD test looks at the amount of oxygen
consumed by living organisms that are feeding off organic matter present in
waste. The test is generally performed in such a manner as similar to
naturalistic conditions as possible, since the test ultimately studies the
pollution of water bodies in everyday conditions.
When the test is performed over a five-day period, it is referred to as a “Five
Day BOD”, or a BOD5.

In order for the test to be quantitative, the samples must be protected from
air to prevent the re-aeration as the dissolved oxygen level diminishes. Also,
since oxygen has limited solubility in water (9 mg/L @ 20 degrees Celsius ), it is
necessary to dilute the wastewater samples as the test is performed.

OBJECTIVES

The objective of this lab is to learn how to perform a BOD5 test of an influent
wastewater collected from the Northwest Mesa Wastewater Treatment Plant.
MATERIALS

• 300 ml BOD bottles

• Large glass or plastic bottle with siphon. Avoid using detergents to clean
these bottles. Periodically clean with bleach water.
• DO meter
• Graduated cylinders or serological pipettes.

Nutrient Solutions:

1. Phosphate buffer
2. Magnesium sulphate solution
3. Calcium chloride solution
4. Ferric Chloride solution
5. Ammonium chloride solution

PROCEDURE

 Preparation of dilution water, Part 1 (Performed by the TA the day

before)
1. Desired volume of DI water was added to 20-L on a plastic 20 L carboy
2. The water was aerated overnight by bubbling in ambient air
compressed air. The saturated O2 concentration in the water should be
around 8.4 mg/L DO.

 Preparation of dilution water Part 2 (Students part)

3. In the overnight aerated dilution water, 1 mL per liter of the five
nutrient solutions was added: phosphate buffered the magnesium
sulphate, calcium chloride, ferric chloride, and ammonium chloride.
4. pH of the dilution water was adjusted to 6.5-7.5.

An influent wastewater sample was provided by the TA for testing.

 Dilution technique:

1. Each BOD bottle was filled by slowly adding sufficient dilution water
with nutrient so that the stopper can be inserted without leaving an air
bubble but not so much that there is overflow
2. In addition to the sample bottles, triplicate bottles were completely
filled with dilution water to be incubated as blanks.
3. Each bottle was labelled carefully, identifying the sample and volume
used.

 Incubation and Dissolved Oxygen (DO) determinations

1. The initial DO of the dilution water blanks was measured and

recorded using the calibrated DO meter.
2. The initial DO in a few of the samples was measured and recorded to
ensure consistency between initial DO in blanks and in inoculated
bottles.
3. All BOD bottles were placed in a cabinet or in a closed box away from
light.
4.Bottles were Incubated at room temperature for 5 days.
5. After 5 days, the DO of the dilution water blanks and the sample
bottles were measured and recorded.
RESULTS

Data table shown below

WW vol DO DO Dilution factor BOD5

(mL) initial Final
0.006666667 615
2 8.2 4.1
0.006666667 675
2 8.3 3.8
0.006666667 600
2 8.9 4.9
0.006666667 705
2 8.9 4.2
0.01 120
3 8 6.8
0.01 190
3 8.3 6.4
0.01 160
3 8.3 6.7
0.01 230
3 8.4 6.1
0.013333333 292.5
4 8.9 5
0.013333333 277.5
4 9.2 5.5
0.013333333 300
4 9.4 5.4
0.016666667 552
5 14.6 5.4
0.016666667 654
5 14.6 3.7
0.016666667 600
5 14.7 4.7
0.016666667 558
5 14.7 5.4
0.025 336
7.5 8.4 0
0.025 244
7.5 8.3 2.2
0.025 280
7.5 8.2 1.2
0.025 332
7.5 8.3 0
 0.033333333 360
10 13 1
0.033333333 294
10 11 1.2
0.033333333 393
10 14.7 1.6
0.041666667 115.2
12.5 7.6 2.8
0.041666667 129.6
12.5 7.8 2.4
0.041666667 156
12.5 7.9 1.4
0.05 116
15 5.8 0
0.05 120
15 6 0
0.05 102
15 5.1 0
0.05 156
15 7.8 0
AVERAGE BOD5= 333.2

WW volume vs DO final
8
7
6
5
DO Final

4
3
2
1
0
2 2 3 3 4 4 5 5 7.5 7.5 10 12.5 12.5 15 15
WW volume(mL)
Calculation:

Formula used are given as

Sample calculation

Sample volume =2 mL

Initial DO =8.2

Final DO=4.1

Dilution factor=2/300 =0.0067

DISCUSSION

From this experiment, the BOD of the wastewater sample analyzed was
determined to be 333.2 mg/L for the solution diluted with influent.

In the test, the observations made did not verify with the theory of the
dissolved oxygen’s depletion over a 5day period. In fact, it was observed that
more titrant was necessary after solutions sat for 5 days, for the influent
samples. As seen from the % depletion of our blank sample of dilution water &
influent sample, we see that oxygen concentration actually increases. This
suggests that an error was present in our experiment. Such possible sources of
error may have risen from the improper sealing of the crucibles, which would
have allowed for air to enter the samples. This causes error in the data since
outside sources of oxygen might contribute to reaeration in the solutions.
Other possible sources of error would include the loss of dissolved oxygen
during transfer from its package to the BOD bottles.
 It was also observed that the stoppers used in this experiment were not
perfectly aligned to fit to the BOD bottles used, causing for an inevitable
bubble to be present in the solution. This might have allowed for air to enter
the solution, hence allowing for outside oxygen to add to the system. To
improve this error, one should have used extra precaution in closing the
system. Such measures of preventing this error include:

a) Sealing the flask with water and leaving the bottles in a closed
fumehood or oven (set at 20 degrees Celsius) for the duration of the 7-
day period.
b) Making sure the bottle stopper is aligned properly or using a vacuum
to leave the systems (bottles) in.

The United states wastewaters regulation of BOD in a wastewater sample is 20

mg/L over a 5-day incubation period. Neglecting the samples we analyzed
which gave inaccurate results, we will only consider the sample diluted with
effluent (treated sample), since it gives reasonable results. The sample was
found to be 7.6 mg/L BOD. Since this falls under the standard accepted BOD
concentration, it could be said that the wastewater sample we analyzed is
acceptable to flow back into the streams after being treated with effluent and
will not be considered a hazard to the environment and aqua life. The sample
containing influent, had the results been accurate according to theory, should
give a result that contains a BOD concentration higher than that which was
treated with effluent, since effluent is a treatment source that feeds the
microorganisms.

It should also be noted that the sample BOD concentration is not completely
precisely accurate in comparison to the United states wastewater standard of
BOD level. This is because the sample that was analyzed was left under a 7-day
incubation period, where as the standard is suggested to be studied after a 5-
day incubation period. For this reason, it should be noted that more time is
given for the sample to be depleted of oxygen, and thus giving a BOD result
that is likely to be .With this said, it is hard to say whether or not the results
obtained from this laboratory experiment provides sufficient data to make an
accurate conclusion for the wastewater sample analyzed.
APPENDICES

APPENDIX A – SUSPENDED SOLIDS DATA

Source: http://angel.cc.umanitoba.ca - Posted November 24th, 2010 by Victor

Wei, lab instructor for CHEM 2560 – Water Quality Analysis for Engineers