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Subject Code/
BFC 32501/ Sem II 20172018
Section 9
1. Muhammad Khairul Faiz Bin Mat Zen (CF160101)
Group Members 2. Mohd Farezy Bin Baharudin (CF160175)
3. Nur Adilah Binti Saiful Bahari (CF160192)
4. Nurlaila Binti Mohd Isa (AF150094)
5. Fatin Syamimi Binti Amirudin (AF150116)
Lecturer /
Instructor/ Tutor Prof. Madya Dr. Radin Maya Saphira Binti Radin
Name Mohamed
Submission Date 20/3/2018

CLO 1: 10%

CLO 2: 15%

Marks CLO 3: 20%

Received Stamp TOTAL

Examiner Comments
Lab Report Assessment Rubrics



Assessed by: Prof. Madya Dr. Radin Maya Saphira Binti Radin Mohamed

Section: 9
Student 1. Muhammad Khairul Faiz Bin Mat Zen
names: 2. Mohd Farezy Bin Baharudin

3. Nur Adilah Binti Saiful Bahari

4. Nurlaila Binti Mohd Isa
5. Fatin Syamimi Binti Amirudin

CLO 2: Organize ideas and method to carry out suitable testing in solving the assigned problems [P4].
Assessment Criteria Sub-criteria Level 1 2 3 4 5 Weight Score
Objective P4 1

Sample location P4 1

Parameter P4 1

Sampling Procedure P4 1
Proposal Perform the proposal
On-site measurement P4 1

Laboratory Procedure P4 5

T otal /50

P ercentage /10%

CLO 1: Prepare rep orts of environmental and transportation tests based on relevant t est standards and compare with t heory [C2].

C2 2

Graphical analysis of result (if needed) C2 2

WQI analysis C2 2

Discussion C2 4
Full Laboratory Writing a full laboratory
Report report Conclusion (purpose suitable treatment) C2 2

Reference (min 10 & cited in report) C2 1

Appendix C2 1

Writing Format C2 1

Total /75

Total /15%
CLO 3: Describe the laboratory testing procedures among group members throughout the experiments. [PLO5, A2]
Assessment Criteria Sub-criteria Level 1 2 3 4 5 Weight Score
Safety attire (PPE) A1 4

Team work: contribution from all members A1 4

Convey information in Follows the testing instructions including A2 4

group safety cautions
Bench organization (during & after test) A2 4

Data collection method (datasheet, etc) A2 4


Total (%)
Assessor signature/ date:

Individual Quiz: student name : 5%

No Student Name Quiz Mark

1. Muhammad Khairul Faiz Bin Mat Zen (CF160101)
2. Mohd Farezy Bin Baharudin (CF160175)
3. Nur Adilah Binti Saiful Bahari (CF160192)
4. Nurlaila Binti Mohd Isa (AF150094)
5. Fatin Syamimi Binti Amirudin (AF150116)
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.


i) Students should 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.

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

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 (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 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 Demand” or 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.


i. 3 units BOD bottles – 300ml
ii. 100ml beaker
iii. 100ml graduated cyclinder
iv. 1unit pipettes (ml)
v. DO meter
vi. pH meter
vii. Air pump
viii. Incubator, capable of maintaining 20 ± 1°C

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 dissolved in distill 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.
BOD Dilution water

Add 1ml of each of the 4 nutrient buffer solutions to 1liter of deionized water and aerate for at least 1 hour
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.



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.

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.
Maximum mL sample = [ ( 8 mg/L – 1 mg/L ) x 300 mL ] / 400 mg/L
Maximum mL sample = ( 7x 300 ) / 400 = 5.24 mL

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 Volume of sample to be diluted

Range to 300 ml in BOD bottle (ml)
(mg O2/L)
0–7 300
7 – 21 100
12 – 42 50
30- 105 20
60 – 210 10
120 - 420 5
300 - 1050 2
600 - 2100 1
1200 - 4200 0.5
3000 - 10500 0.2
6000 - 21000 0.1

To determine the value of the BOD in mg/l, use the following formula:
When dilution water is not seeded:
D 1 – D2
BOD5, mg/L =
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 wastewater typically has a BOD of around
300 mgO2/l].

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. 50 mL of samples is poured into a 100 mL beaker.

Figure 1: 50mL of sample

2. The pH of the sample is measured 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

Figure 2: Measure the pH sample

3. 1 N sulfuric acid is added if the sample is alkaline, or 1N sodium hydroxide if the sample
is acidic, until the pH reaches 7.0.

4. The amount of sulfuric acid or sodium hydroxide needed is calculated to neutralize 1000ml
of the sample.

5. The calculated amount of acid or base is added to the sample.

6. Steps 1-5 is repeated until the pH test shows pH 7.0.

7. The amount of 1 N sodium hydroxide or 1 N sulfuric acid needed to neutralized the sample
to pH 7.0 is calculated 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

1. 3 unit of 300ml BOD bottles (1 blank, 2 samples) is labeled separately.

2. The volumes is pipetted appropriately into bottles. All the bottles is filled to the top with dilution

Figure 3: Pipetted the volume Figure 4: Filled the bottle

3. The initial DO concentration is measured using DO meter.

Figure 5: Measure the initial DO

4. All the bottles is placed into the BOD incubator, this is set at 20oC and is dark to prevent the
growth of algae. Leave for 7 days.

5. After 7 days, the samples is removed from the incubator and the final DO content is measured
for 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.

Sample: pH : 6.65
Source : Drainage in front of FKAAS building

BOD Data table.

Date / Time Started: 6/3/2018 (2.00 PM)

Volume of
Volume of Initial DO Final DO
BOD ID Dilution Water
Sample (mL) (mg/L) (mg/L)

Sample 1 4 150 8.78 7.61

16 150 8.77 7.46

Blank 1 0 150 8.78 8.32


1. Calculation of BOD

D1 – D2
BOD5, mg/L =

i. Sample 1 :
(4 ÷150)
: 43.88 mg/L

ii. Sample 2 :
: 12.28 mg/L
2. Does your BOD dilution water show ’toxic effect’?

Both sample does not show any toxic effect. This is because the waste water is took from the drainage near
the FKAAS building is not from a toxic source. The pH value of the sample is nearly neutraland after 4
mL of wastewater is used. The value of BOD obtained is slightly higher than the sample size of 16 mL. It
means that the sample does not show any toxic effect.

3. Could you rely on your BOD results? Why?

No. This is because some errors were occurred. The dilution water was prepared and stored for some times
without proper quality control before the experiment. This caused growth of some biological that cannot
be seen by naked eyes. Existence of biological in the dilution water caused inaccuracy of the result.
Moreover, the waste water used in this experiment does not undergo pretreatment process. It may contain
dissolved heavy metal. The presence of heavy metal make the result obtained may be different.

Based on the experiment, the BOD for both sample are not polluted as the BOD is below 100mg/L. It is
not in the range of Standard A and Standard B. Standard A (20mg/L) is the indicator for drinking water
while Standard B (50mg/L) is the indicator for inland water quality. Therefore, the water sample that
located near the FKAAS building is not polluted. This is because there are no activities that occurs near
the drain and there is no waste product that can be pollute the water inside the drain.

DO levels are highest if the sample of water is not polluted means there is no toxic and bacterial
effect so in sample of water there is a lot of oxygen demanding wastes is well within self- purification
capacity. Otherwise, the BOD will low which is water sample not have microbes live. Then DO levels low
if sample water not have enough oxygen to dissolve in water that need to all leaving things.


From the experiment, it can be concluded that the objective of this experiment (Biochemical Oxygen
Demand) has been achieved. The purpose of this experiment is to measure the amount of dissolved oxygen
in the water for a specific period time and temperature. The sample of water source which is located near
the FKAAS building is not polluted. The results of the BOD which is 43.88mg/L ( sample 4 ml ) and
12.28mg/L ( sample 16 ml ) is still in the range of Standard A and Standard B. However,
this waste water cannot simply discharge to the river untreated. It must be treating first by any possible
treatment to make sure that water is safe and will not to be polluted to the river.