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FACULTY : CIVIL &

ENVIRONMENTAL ENG.
DEPART : WATER &
ENVIRONMENTAL ENG.
LAB : ENVIRONMENTAL
ENGINEERING
EXPERIMENT :BIOCHEMICAL
OXYGEN DEMAND
(BOD)

EDITION:
REVISION
02
NO:
EFFECTIVE 28/12/20
DATE: 15
AMENDMENT
DATE:

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)
Studentsshould be able to describe the importance of BOD in the
environmental studies.
ii)
Students should 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 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.

FACULTY : CIVIL &


ENVIRONMENTAL ENG.
DEPART : WATER &
ENVIRONMENTAL ENG.
LAB : ENVIRONMENTAL
ENGINEERING
EXPERIMENT :BIOCHEMICAL
OXYGEN DEMAND
(BOD)

EDITION:
REVISION
02
NO:
EFFECTIVE 28/12/20
DATE: 15
AMENDMENT
DATE:

Sometimes it is advantageous to measure just the oxygen demand exerted


by organic
(carbonaceous) compounds, excluding the oxygen demand exerted 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
Demandor BOD.
Generally, high BOD indicates a high content of easily degradable organic
material in sample, and low BOD indicates a low volume of organic material
substances which are difficult to break down.
4.0 EQUIPMENT & REAGENTS

i.
ii.
iii.
iv.
v.
vi.
vii.
viii.

6 units BOD bottles 300ml


100ml beaker
100ml graduated cyclinder
1unit pipettes (ml)
DO meter
pH meter
Air pump
Incubator, capable of maintaining 20 1C

Nutrient Buffer solution


1. Phosphate Buffer
- Dissolve 8.5g potassium di hydrogen phosphate (KH2PO4), are dissolved in
approximately 500ml distilled water, 21.75g di potassium hydrogen
phosphate (K2HPO4) are added, followed by 33.4g di sodium hydrogen
phosphate (Na2HPO4) and 1.7g ammomium chloride (NH4CI). Adjust pH to
7.2 if necessary with either 1 NH2SO4 or NaOH. Dilute to 1liter
2. Magnesium sulfate
- 22.5g magnesium sulphate (MgSO4.7H2O) is diddolved in distrilled water
and the solution made up to 1liter.
3. Calcium chloride
- 27.5g of anhydrous calcium chloride (CaCI2) are dissolved in distilled water
and the solution made up to 1liter.
4. Ferric chloride
0.25g FeCI3.6H2O is dissolved in distilled water and the solution made
up to 1liter.

FACULTY : CIVIL &


ENVIRONMENTAL ENG.
DEPART : WATER &
ENVIRONMENTAL ENG.
LAB : ENVIRONMENTAL
ENGINEERING

EDITION:
REVISION
02
NO:
EFFECTIVE 28/12/20
DATE:
15

EXPERIMENT :BIOCHEMICAL
OXYGEN DEMAND
(BOD)

AMENDMENT
DATE:

BOD Dilution water


Add 1ml of each of the 4 nutrient buffer solutions to 1liter of deionized
water and aerate for at least 1 hr before conducting BOD test to ensure
the DO concentration in the dilution water is at least 7.5 mg/L.
Chemical for pH adjustment
1N of sulfuric acid, H2SO4 or 1N sodium hydroxide, NaOH.
5.0 BOD5 MEASUREMENT PROCEDURES
5.1 DETERMINATION OF SAMPLE SIZE :
Once a general range for the BOD of a sample has been determined, the
dilutions can be established which will ensure that at least one dilution will
meet the criteria for valid BOD results. The following procedure can be used
to calculate volumes for sample dilution from the estimated BOD.
For example, 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:
A. 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
B.

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
FACULTY : CIVIL &
ENVIRONMENTAL ENG.
DEPART : WATER &
ENVIRONMENTAL ENG.
LAB : ENVIRONMENTAL
ENGINEERING

EDITION:
REVISION
02
NO:
EFFECTIVE 28/12/20
DATE:
15

EXPERIMENT :BIOCHEMICAL
OXYGEN DEMAND
(BOD)

AMENDMENT
DATE:

NOTE: Those sample dilutions which deplete less 2 mg/L, or have a final DO of
less than 1 mg/L would not be use in the calculation of the average sample
BOD.
Table 1: Volume of sample for dilution
Expected BOD
Range
(mg O2/L)
0-7
7 - 21
12 - 42
30- 105
60 - 210
120 - 420
300 - 1050
600 - 2100
1200 - 4200
3000 - 10500
6000 - 21000

Volume of sample to be diluted to


300 ml in BOD bottle (ml)
300
100
50
20
10
5
2
1
0.5
0.2
0.1

To determine the value of the BOD in mg/l, use the following formula:

When dilution water is not seeded:

D1 D 2
BOD5,mg/L =
P

D1 = DO of diluted sample immediately after preparation, mg/L,


D2 = DO of diluted sample after 5 d incubation at 20oC, mg/L,
P = reciprocal of dilution factor, D= volume of BOD bottle/volume of sample used in
mL

NOTE: The samples must undergo dilution if they have a BOD > 7, otherwise all the
dissolved oxygen will be used up before 5 days have elapsed. [Note: raw domestic

FACULTY : CIVIL &


ENVIRONMENTAL ENG.
DEPART : WATER &
ENVIRONMENTAL ENG.
LAB : ENVIRONMENTAL
ENGINEERING
EXPERIMENT :BIOCHEMICAL
OXYGEN DEMAND
(BOD)

EDITION:
REVISION
02
NO:
EFFECTIVE 28/12/20
DATE: 15
AMENDMENT
DATE:

5.2 PRE-TREATMENT SAMPLES.


1) That contain caustic alkalinity or acidity
2) Caustic alkalinity or acidity can prevent bacteria from growing during the
course of the BOD test.
3) 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 test is
performed.
NOTE :Neutralized samples must be seeded for the BOD test.
Procedure for neutralizing samples:
1.
2.
3.
4.
5.
6.
7.

Pour 50 mL of samples into a 100 mL beaker.


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 countinue with steps 3 6, otherwise perform
the BOD test on the untreated sample.
Add 1 N sulfiris acid if the sample is alkaline, or 1N sodium hydroxide
if the sample is acidic, intil the pH reaches 7.0.
Calculate the amount of sulfuric acid or sodium hydroxide needed to
neutralize 1000ml of the sample.
Add the calculated amount of acid or base to the sample.
Repeat steps 1-5 until the pH test shows pH 7.0.
Calculation the amount of 1 N sodium hydroxide or 1 N sulfuric acid
needed to neutralized the sample to pH 7.0 using the following formula:
mL needed = ( mL acid or base used x mL total test sample ) mL sample
portion used for neutralization.
For example, suppose 1.3 mL of 1 N NaOH are use to neutralize 50 mL of
sample to pH 7.0 calculate the volume of NaOH to be added to neutralize
the sample as follows:
mL 1 N NaOH needed = ( 1.3 mL x 1000 mL ) /50 mL = 1300 / 50 = 26ml

FACULTY : CIVIL &


ENVIRONMENTAL ENG.
DEPART : WATER &
ENVIRONMENTAL ENG.
LAB : ENVIRONMENTAL
ENGINEERING
EXPERIMENT :BIOCHEMICAL
OXYGEN DEMAND
(BOD)

EDITION:
REVISION
02
NO:
EFFECTIVE 28/12/20
DATE: 15
AMENDMENT
DATE:

5.3 BOD5PROCEDURES
1.
2.
3.
4.

Label separately 4 unit of 300ml BOD bottles (1 blank, 3 samples)


Pipette appropriate volumes into bottles. Fill all the bottles to the top with dilution water.
Measure the initial DO concentration using DO meter.
Place all the bottles into the BOD incubator, this is set at 20oC and is dark to prevent the
growth of algae. Leave for 5 days.
5. After 5 days, remove the samples from the incubator and measure the final DO content of
each.
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 NOT be less than
0.2 mg/l. in any event, do not use the depletion obtained as blank correction.

6.0 RESULT AND CALCULATION


a)
b)
c)
d)
e)
f)

Type of sample:
Initial pH sample:
Adjustment pH sample:
Volume acid used:
Volume base used:
Volume of sample used:

FACULTY : CIVIL &


ENVIRONMENTAL ENG.

.
.
.
.
.
.

EDITION:

DEPART : WATER &


ENVIRONMENTAL ENG.
LAB : ENVIRONMENTAL
ENGINEERING
EXPERIMENT :BIOCHEMICAL
OXYGEN DEMAND
(BOD)
Volume
of
incubati
Sample
on
bottle,
ml
BLANK
300
1
300
2
300
3
300
7.0 DATA ANALYSIS

Volume
of
sample
size, ml
0
50
50
50

Initial
DO,
mg/l
7.48
7.46
7.46
7.47

REVISION
02
NO:
EFFECTIVE 28/12/20
DATE: 15
AMENDMENT
DATE:

Final
DO,
mg/l

DO
depletio
n, mg/l

BODx,
mg/l

4.6
4.6

i.

Show all the calculation and state if any of the data needs to be discarded.

ii.

Does your BOD dilution watershow toxic effect?

iii.

Could you rely on your BOD results? Why?

iv.

Determine the pollution level of samples based on the BOD results.

v.

Differentiate between seeded and unseeded BOD.

8.0 DISCUSSIONS

PREPARED BY:

SIGNATURE:

POSITION:
DATE:

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