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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.
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Hazardous Material Safety:
1. Unauthorized experiments are prohibited. Perform the experiments as directed by
your instructor.
2. Never take chemicals, supplies, or equipment out of the laboratory.
3. Students are not allowed to enter the Chemical Stockrooms.
4. 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. Learn the locations and operation of emergency equipment. This includes eyewash,
fire extinguishers, etc.
2. Know what to do in case of an emergency.
3. Know how to exit the building in case of an emergency.
4. Report ALL accidents, injuries, and near misses (close-calls) to your instructor
immediately
5. Treat burns immediately by putting the burned area under cold water for at least 15
minutes.
6. 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. 4
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

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MA01 Total Suspended Solid (TSS):
Weigh the Pour 15 mL of water
Switch on record the sample ( from UTHM
the initial dry lake) on the filter
balance weight of paper and leave it Take the filter paper,
( at least filter until the filtration place on the
30 min paper finishes evaporating dish
before the
test)

Weigh
and
record the Place the
final dry evaporating dish
weight of to cool at room Place the
filter temperature in evaporating disk
paper dessicator inside the drying
oven at 1050C for at
least 1 hour or until
completely dry.

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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 W2


residue (g)
Weight of residue (g) W

Volume
2) In of sample
your opinion, (mL) to discharge the sample
is it possible V used in this test to be
discharged to the river?. Give your reason.

B) Discussion

3) Why is the impacts of total suspended solid on health of the river? 7


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

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MA02 Bacteria Count
Note Prepare freshly by
All equipment boiling the mixture of:
used in this peptone = 5 g
experiments Inoculate
plate
Beef extract = 3 g
should be containin Agar = 15 g
sterilized in an g Distilled water = 600
autoclave at nutrient mL
agar
1210C for 15 min
Sterile the spreader
Carefull with
each time before
ethanol and
spread the bacteria
Bunsen burner
sample over the agar
Close the plate. by immerse the
Invert the plates spreader in ethanol,
and incubate for 24 tap on the side of the
hours at 300C beaker to remove the
excess ethanol and
After being flame
Close the plate.
incubated for 1 da, Invert the plates
take out the plates and incubate for 24
and count the hours at 300C
colony using colony
counter
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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
2) Between pour plate and spread plate,which method is more flexible to be used as
enumeration procedure for various types of bacteria ? 11
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
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MA03 Biochemical Oxygen Demand (BOD)
1. Collect water from UTHM lake
2. Measure the pH-
i) 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 Dilution water Sample Total


Bottle ID ( freshly prepared) volume
Sample 1 297 3 300
Sample 2 297 3 300
Sample 3 297 3 300
Blank 1 300 0 300
Blank 2 300 0 300
Dilution water * = Distilled water + phosphate buffer + magnesium sulfate + calcium Chloride + Ferric
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* Read 5210 B (Standard Method) for details
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 5 days at 20 0C
(typically), or 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 ( Eqn C)


P
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

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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 Volume of Dilution Initial DO Final DO (mg/L)
sample (mL) water ( mL) (mg/L)
Sample
1
Sample
2
Sample
3

2) In1your opinion, is it possible to discharge the sample used in this test to be


Blank

discharged to the river?. Give your reason.


Blank 2

B) Discussion

3) Why BOD test should be done promptly after the collection? 17


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:
Blank Vial ( do it in fume Sample vial ( do it in fume
hood) hood)
- 1.5 mL distilled water - 1.5 mL sample
- 2.5 mL standard potassium - 2.5 mL standard potassium
dichromate dichromate
solution ( 0.04167 M) solution ( 0.04167 M)
- 1.5 mL sulfuric acid reagent - 1.5 mL sulfuric acid reagent COD Vial
3. Close the cap tightly
4. Mix the mixture by shaking side by side
5. Put the COD vial into the COD reactor and digest at OC for hour
6. Take out the COD vial from the COD reactor, leave to cool at room temperature
7. Titration using standard ferrous ammonium sulfate (FAS) is crucial for COD
determination because:

COD ( mg/L) = B S X D x 1000


Note, this procedure had amended
T
to make it possible within 2 hours
where; Refer Standard Method 5220 for
B = titre for blank (mL) details of COD measurement.
S = titre for sample (ML)
T = titre (mL)
D = 1 if sample is not diluted COD Reactor 21
MA05 Chemical Oxygen Demand (COD):
To obtain T To obtain S To obtain B
(mL) (mL) (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:


In a flask of 100 mL:
100 mL flask
Mixed thoroughly 1.5 Pour the COD vial
Pour the COD vial
mL standard potassium (sample)
(sample)
dichromate solution Fill up the COD vial
Fill up the COD vial
( 0.04167 M) with 15 with distilled water for
with distilled water for
mL distilled water rinsing. Pour the water
rinsing. Pour the water
to the flask. Repeat it
to the flask. Repeat it
Then add 3.5 ml sulfuric twice.
twice.
acid reagent very slowly
Then add 2 drops of
Then add 2 drops of
Then add 2 drops of ferroin indicator
ferroin indicator
ferroin indicator
Then titrate using FAS
Then titrate using FAS
Then titrate using FAS (as titrant until the
(as titrant until the
(as titrant until the solution turns RED
solution turns
solution turns RED
Record the titrant used RED
Record the titrant used as S
as T
Record the titrant used
Note, this procedure had amended to make it possible within
as B
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2 hours Refer Standard Method 5220 for details of COD
MA05 Chemical Oxygen Demand (COD):

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

3) 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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