BFC 32501

ENVIRONMENTAL ENGINEERING LABORATORY

SECTION 2

SEM 1 2006/2017

PREPARED BY
HJH ROSLINDA SESWOYA

General Requirements
1. Be properly prepared to do the experiment scheduled for that day.
2. There will be no smoking, using CELL PHONES, drinking, chewing gum, or eating
in the laboratory.
3. Do not touch your face, handle contact lenses, apply cosmetics or put your
fingers in your mouth.
4. Always wash your hands after handling viable material.
5. Avoid working alone in the laboratory.
6. Horseplay will not be tolerated. Avoid distracting others that are working.
7. At the end of each lab session, clean your lab bench, put away all materials,
and wash your hands thoroughly before leaving the laboratory. Wastes should
be deposited in appropriate receptacles.
8. Gloves and lab coats are to be removed prior to leaving the laboratory and may
not be worn in non-laboratory areas.
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Personal Protective Equipment (PPE):

1. Observe all laboratory signs and chemical labels for required protective
equipment.
2. Safety glasses or chemical splash goggles will be worn at all times when
performing laboratory work.
3. Wearing contact lenses is discouraged.
4. Always wear gloves when handling any chemicals or microorganisms.
5. You are required to wear laboratory coats while working to prevent
contaminating your clothes.
6. Wear shoes that cover the entire foot; NO sandals or open shoes.
7. Tie back long hair and confine loose clothing to keep away from flames
and chemicals.

Hazardous Material Safety:

1.
2.
3.
4.

Unauthorized experiments are prohibited. Perform the experiments as directed by your instructor.
Never take chemicals, supplies, or equipment out of the laboratory.
Students are not allowed to enter the Chemical Stockrooms.
Know where the MSDS (Material Safety Data Sheets) are for your chemicals. MSDS contain information
that you will need in case there is an emergency.
5. Never taste or smell a chemical.

Spills and Other Lab Incidents:

1.
2.
3.
4.
5.
6.

Learn the locations and operation of emergency equipment. This includes eyewash, fire extinguishers, etc.
Know what to do in case of an emergency.
Know how to exit the building in case of an emergency.
Report ALL accidents, injuries, and near misses (close-calls) to your instructor immediately
Treat burns immediately by putting the burned area under cold water for at least 15 minutes.
Clean up all spills immediately and notify your instructor. This includes water. In the event that a chemical
is spilled/splashed on your eyes, skin or body, IMMEDIATELY rinse the affected area for 15 minutes.
7. If your skin is exposed to a viable material, wash the affected area with antibacterial soap and hot water.
8. In case of accidental ingestion, notify instructor immediately and call for emergency medical services
(Dial 999).
9. Have the MSDS ready for personnel to determine the necessary course of treatment.
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MA01

Total Suspended Solid (TSS):

The term total suspended solids can be referred to materials which

are not dissolved in water and non filterable in nature.
It is defined as residue upon evaporation of non-filterable sample
on a filter paper

Total Suspended Solid (TSS):

MA01

Weigh the
record the
initial
dry
weight
of
filter paper

Switch on
the balance
( at least 30
min before
the test)

Weigh and
record the
final
dry
weight
of
filter paper

Pour 15 mL of water
sample ( from UTHM
lake) on the filter paper
and leave it until the
filtration finishes

Place the evaporating

dish to cool at room
temperature
in
dessicator

Take the filter paper,

place on the evaporating
dish

Place the evaporating

disk inside the drying
oven at 1050C for at least
1 hour
or until
completely dry.

MA01

Total Suspended Solid (TSS):

For Environmental Engineering Laboratory ( BFC 32501) SEM 1 2016/2017, submit
the lab report by answering the following
A )Data collection and analysis
1) Complete the table, What is the TSS of the sample? Show the calculation.
Description

Weight (g)

Weight of the clean filter paper (g)

W1

Weight of the filter paper and the residue (g)

W2

Weight of residue (g)

Volume of sample (mL)

2) In your opinion, is it possible to discharge the sample used in this test to be

discharged to the river?. Give your reason.
B) Discussion

1) Why is the impacts of total suspended solid on health of the river?

MA02

Bacteria Count

A plate count reflects the number of viable microbes and assumes

that each bacterium grows into a single colony.
Because it is impossible to say that each colony actually arose from
an individual cell, plate counts are reported as the number of
colony forming units (CFU) instead of the number of cell.
A plate count may be done on plates prepared by either the pour
plate method or the spread plate method.
To insure a countable plate, a series of dilutions ( as shown below)
should be plated. The serial dilution should give at least one
countable plate ( For the bacterial count , apply the similar dilution
and the numbers of plate as shown below)
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MA02

Bacteria Count

MA02
Note
All equipment used
in this experiments
should be sterilized
in an autoclave at
1210C for 15 min
Carefull with ethanol
and Bunsen burner

Bacteria Count
Inoculate
plate
containing
nutrient agar

Prepare freshly by boiling the

mixture of:
peptone = 5 g
Beef extract = 3 g
Agar = 15 g
Distilled water = 600 mL

Close the plate. Invert

the plates and incubate
for 24 hours at 300C

After being incubated

for 1 da, take out the
plates and count the
colony using colony
counter

Close the plate. Invert

the plates and incubate
for 24 hours at 300C

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MA02

Bacteria Count

For Environmental Engineering Laboratory ( BFC 32501) SEM 1 2016/2017, submit the
lab report by answering the following
A )Data collection and analysis
1) Complete the table, What is the bacterial count of the sample? Show the value in unit
of bacteria/mL sample.
Bacterial count = Number of colony/ (dilution x volume of sample applied on plate)
Dilution

Count

Countable ( Yes/No)

1/10
1/100
1/1000
1/10000
1/100000
B) Discussion

1) Between pour plate and spread plate,which method is more flexible to be used as
enumeration procedure for various types of bacteria ?

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MA03 Biochemical Oxygen Demand (BOD)

It is a measure of the quantity of oxygen
used by microorganisms (eg.aerobic
bacteria) in the oxidation of organic matter.
In other words: BOD measures the change
in dissolved oxygen concentration caused
by the microorganisms as they degrade the
organic matter.
High BOD is an indication of poor water
quality

12

MA03 Biochemical Oxygen Demand (BOD)

1. Collect water from UTHM lake
2. Measure the pHi) if the pH is in the range of 6.5 to 7.5- proceed to Step 3
step 3
ii) if the pH of sample is less than 6.5, then use base to improve the
pH to 7, refer to standard method for detail procedure Seeded BOD
iii) if the pH of sample is bigger than 7.5, then used acid to improve the pH to
pH to 7, refer to standard method for detail procedure Seeded BOD
3. Using the details as below, determine the sample size using Eqn A and
Eqn B
a) Based on the literature, the BOD of A = 400 mg/L ( subject to changed )
b) Assume the DO of saturated dilution water = 8 mg/L
c) The DO depletion at the end of 5 days incubation = 2 mg/L
d) The residual DO = 1 mg/L
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MA03 Biochemical Oxygen Demand (BOD)

Eqn A = minimum waste to be filled in BOD bottle
= [(2 mg/L ) X 300 mL] / 400 mg/L
= 1.5 mL
Eqn B = maximum waste to be filled in BOD bottle
= [(8 mg/L- 1 mg/L ) X 300 mL] / 400 mg/L
= 5.2 mL

Therefore sample size = 1.5 to 5.2 mL (Let say choose 3 mL)

4. Take 5 BOD bottles. Use 3 bottles for sample, and 2 for blank
5. Fill the BOD bottles ( one by one) as below:

BOD Bottle ID

Dilution water
( freshly prepared)

Sample

Total
volume

Sample 1

297

300

Sample 2

297

300

Sample 3

297

300

Blank 1

300

300

Blank 2

300

300

Dilution water * = Distilled water + phosphate buffer + magnesium sulfate + calcium Chloride + Ferric chloride
* Read 5210 B (Standard Method) for details

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MA03 Biochemical Oxygen Demand (BOD)

6. Knock the bottle to eliminate the air bubbles from
the BOD bottle.
7. Measure the DO using DO mete from each samples
and blanks. Record the value as initial DO.
8. Slowly refill the dilution water
( if needed) to the BOD bottles and put the lid.
9. Repeat step 6 to 8 for another BOD bottle.
10. Store the BOD bottles in the BOD incubator for
3 days at 300C
11. On day five/three of incubation days, take out the BOD bottles ,
leave it until it reach the room temperature
12. Read the DO from each samples and blanks. Record the value
as final DO
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MA03 Biochemical Oxygen Demand (BOD)

13. Apply Eqn C to measure BODX for the sample.
BODX mg/L = Di Df
P

( Eqn C)

Where
Di = initial DO ( mg/L)
Df = final DO (mg/L) , 5 days or 3 days
P = Volume of sample used / Total volume

Note:
1)
DO in blanks should not deplete more than 0.2 mg/L
2)
DO in sample should not less than 2 mg/L
3)
Final DO in the samples should not less than 1 mg/L

16

MA03 Biochemical Oxygen Demand (BOD)

For Environmental Engineering Laboratory ( BFC 32501) SEM 1 2016/2017, submit
the lab report by answering the following
A )Data collection and analysis
1) Complete the table, What is the BOD of the sample? Provide the value in
avg std-dev. Show the calculation.
BOD ID

Volume of sample (mL)

Volume of Dilution water ( mL)

Initial DO (mg/L)

Final DO (mg/L)

Sample 1
Sample 2
Sample 3
Blank 1
Blank 2

2) In your opinion, is it possible to discharge the sample used in this test to be

discharged to the river?. Give your reason.
B) Discussion

1) Why BOD test should be done promptly after the collection?

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MA04

JAR Test

This experiment is selected to be done by

students ( in a group) to meet the criteria of open
ended approach
Students (in a group) are required to design and
write the lab sheet on how to conduct the JAR
test.
Before execute the JAR test , students ( in a
group) should present and discuss the lab sheet
with the Lecturer a week before the execution.

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MA05

Chemical Oxygen Demand (COD)

It is based on the fact that all organic compounds,
with a few exception, can be oxidized by the
action of strong oxidizing agents under acid
condition (=Value usually about 1.25 times BOD)
Chemical oxygen demand is to measure the
oxygen requirement of a sample that is likely to
oxidation by strong chemical oxidant ( potassium
dichromate).

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MA05

Chemical Oxygen Demand (COD)

Organic matter in aqueous samples could be
determined by oxidation with dichromate.
The amount of O2 that is chemically equivalent to the
dichromate consumed is defined as the COD of the
sample.
During the oxidation in which sample is heated with
the known excess of dichromate, organic matter is
converted to carbon dioxide and water while
dichromate is reduced to Cr 3+.
The excess dichromate is determined by means of an
oxidation-reduction titration with ferrous ammonium
sulfate (FAS).
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MA05 Chemical Oxygen Demand (COD):

1. Take sample from UTHM lake
2. Take a COD Vial ( 2 pcs), and fill the vial with the following:

3.
4.
5.
6.
7.

Blank Vial ( do it in fume hood)

Sample vial ( do it in fume hood)

- 1.5 mL distilled water

- 2.5 mL standard potassium dichromate
solution ( 0.04167 M)
- 1.5 mL sulfuric acid reagent

- 1.5 mL sample
- 2.5 mL standard potassium dichromate
solution ( 0.04167 M)
- 1.5 mL sulfuric acid reagent
COD Vial

Close the cap tightly

Mix the mixture by shaking side by side
Put the COD vial into the COD reactor and digest at 150 OC for 1 hour
Take out the COD vial from the COD reactor, leave to cool at room temperature
Titration using standard ferrous ammonium sulfate (FAS) is crucial for COD determination
because:
COD ( mg/L) = B S X D x 1000
T
where;
B = titre for blank (mL)
S = titre for sample (ML)
T = titre (mL)
D = 1 if sample is not diluted

Note, this procedure had amended to

make it possible within 2 hours Refer
Standard Method 5220 for details of COD
measurement.

COD Reactor

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MA05 Chemical Oxygen Demand (COD):

To obtain T (mL)

To obtain S (mL)

To obtain B (mL)

Do it in fume hood

Do it in fume hood

Do it in fume hood

In a flask of 100 mL:

In a flask of 100 mL:

Mixed thoroughly 1.5 mL

standard potassium
dichromate solution
( 0.04167 M) with 15 mL
distilled water

Pour the COD vial (sample)

Then add 3.5 ml sulfuric acid

reagent very slowly

Then add 2 drops of ferroin

indicator
Then titrate using FAS (as
titrant until the solution turns
RED

In a flask of 100 mL:

Pour the COD vial (sample)

100 mL flask

Then add 2 drops of ferroin

indicator
Then titrate using FAS (as
titrant until the solution turns
RED

Then add 2 drops of ferroin

indicator

Record the titrant used as S

Record the titrant used as B

Then titrate using FAS (as

titrant until the solution turns
RED
Record the titrant used as T

Note, this procedure had amended to make it possible within 2 hours Refer
Standard Method 5220 for details of COD measurement.

22

Chemical
Oxygen
Demand
(COD):
MA05
For Environmental Engineering Laboratory ( BFC 32501) SEM 1 2016/2017,
submit the lab report by answering the following
Data collection and analysis
1) What is the COD of the sample ? Provide the calculation.
2) In your opinion, is it possible to discharge the sample used in this test to be
discharged to the river?. Give your reason.
Discussion

1) Is it possible to predict the BOD of the sample using the COD value? If yes,
explain how to do that.

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GOOD LUCK !!!

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