OD/CED: 84/85

Semester: Fall-2018

PRACTICAL BOOK
Public Health Engineering Laboratory

Name of Student: __________________________

PA/TC/NC/PC No: ______________

Civil Engineering Wing

Military College of Engineering
National university of Science and Technology
No. CLO PLO Assessment Learning Level of
Practical
No. No. Methodology Domain Learning
1 a. Preparation of Solution
2 b. Determination of pH/ temperature
3 c. Determination of turbidity
4 d. Determination of color
5 e. Determination of TS, TDS and TSS using gravimetric method
6 f. Determination of TFS and TVS using gravimetric method
7 g. Determination of chlorides
Laboratory
8 h. Determination of sulfates using spectrophotometer 3
1,2,3 1,2,3 Reports, Quiz, Psychomotor
9 i. Determination of nitrates using spectrophotometer
Viva
10j. Determination of total hardness
11k. Determination of alkalinity
12l. Determination of DO
13m. Determination of BOD
14n. Determination of COD
15o. Determination of Total Coliform
16p. Mini Project 4
 Table of Contents

S.No Description of Practical Page No.

1. Preparation of solutions 2
2. Measurement of pH/ Temperature value (Electrometric method) 3
3. Determination of Turbidity (Nephlometric method) 4
4. Determination of Color (Platinum-Cobalt method) 5
5. Determination of TS, TDS, TSS, TFS, TVS using Gravimetric method. 6
6. Determination of Chlorides 9
7. Determination of Sulfates using Spectrophotometer 11
8. Determination of Nitrates using Spectrophotometer 12
9. Determination of Phosphate using Spectrophotometer 13
10. Determination of Total Hardness (EDTA Titration method) 14
11. Determination of Dissolved Oxygen 15
12. Determination of Biochemical Oxygen Demand 16
13. Determination Of Chemical Oxygen Demand 20
14. Determination of Total Coliform (Membrane Filtration Method) 22
 2

PREPARATION OF SOLUTIONS

Preparation of Solution:
In volumetric analysis one of the required solutions must be standard, by definition a
standard solution is one whose strength or reacting value per unit volume is known.

Requirements:
Molar or Normal Solution (desired conc.)
Volume required / Quantity

Theory (The solute may be solid or liquid):

If the solute is solid:
Solute (g) = [Molarity x Molecular. wt of solute (g) x Volume (ml)] / 1000
OR
Solute (g) = [Normality x Equivalent .wt of solute (g) x Volume (ml)] / 1000

Laboratory Problem:
Determine the amount of NaHCO3, required to prepare 250ml solution of 0.1M?

Answer:

Comments:
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(Signatures Laboratory Demonstrator with date)

 3

MEASUREMENT OF pH/ TEMPERATURE OF WATER/WASTEWATER

(Electrometric Method)

Apparatus: pH Meter, Beaker, Sample of Water/ Wastewater

Measurement of pH and Temperature

 Connect the pH electrode and ATC/temp probe to the unit.
 Press MODE key repeatedly until the unit displays pH mode
 Rinse the pH electrode with distilled water and immerse it in the sample.
 Allow sufficient time for the display to stabilize.
 The unit will display pH and temperature value of the sample.

Observations/Results:

Sample pH Value Temperature oC

1
2
3

Comments:
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(Signatures Laboratory Demonstrator with date)

 4

DETERMINATION OF TURBIDITY OF WATER SAMPLE

(Nephlometric Method)

Apparatus: Turbidity meter, cleaning cloth,

Procedure:
 Turn the instrument on. Allow it to warm up as per the manufacturer’s
instructions.
 Fill a sample cell to the line from the sample and cap the cell.
 Wipe the cell with a soft tissue to remove water spots and fingerprints on it.
 Slowly invert the sample cell 2-3 times. But do not shake/invert rapidly.
Otherwise, entrapped air bubbles and false/incorrect readings could result.
 Place the sample cell in the instrument cell compartment, as shown and close the
lid.
 Record the stable reading

Observations/Results:

Sample Turbidity NTU/ FTU

1
2
3

Comments:
__________________________________________________________________________________________________
__________________________________________________________________________________________________
___________________________________________

(Signatures Laboratory Demonstrator with date)

 5

DETERMINATION OF COLOR OF WATER/WASTEWATER SAMPLE

(Colorimetric Method)

Apparatus/Chemicals: Spectrophotometer, filter paper, filtration assembly etc

Procedure:

 Filter 50 ml of deionized water through Vacuum pump assembly.

 Fill cell with 25 ml of filtered deionized water (Blank).

 Filter 50 ml of sample water through vacuum pump assembly.

 Fill another cell with 25 ml of filtered sample water.

 Enter the stored number for True color e.g press 120.

 Dial wavelength to 455 nm

 Place the blank and close the light shield.

 Press ZERO buttons, the display will show zeroing ….. And then 0 UNITS PtCo
APHA.

 Place the sample cell and close the light shield.

 Press READ and display will show READING …. And then result will be display.

Observations/Results:

Sample PtCo
1
2
3

Comments
__________________________________________________________________________________________________
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(Signatures Laboratory Demonstrator with date)

 6

DETERMINATION OF SOLIDS (TS, TSS, TDS, TFS, TVS)

(Gravimetric Method)

Apparatus: filter paper, china dish, weighting balance, oven etc

Procedure for Total Solids (TS):

 A well-mixed sample (say 50ml) is evaporated in a weight dish and dried at
constant weight at 103C to 105C for I hr in Oven or muffle Furnace.
 Cool, desiccate and weight, (say B mg)

Calculation:
mg/L of Total Solids = [(A - B) x 1000]/ml of sample
Where; A= weight of dried residue +dish (mg)
B = weight of dish, only (mg)

Procedure for Total Dissolved Solids (TDS) and Total Suspended Solids (TSS):
Total Suspended Solids (TSS)
 Wash the filter with distilled water and dry it an oven at 103±2 Co for 1.0 hour
and weigh the filter (say A1 mg).
 Ignite an evaporating dish in a muffle furnace at 550±5 Co for 1.0 hour and weigh
the dish (say A2 mg).
 Now filter the sample through the filtration assembly, use vacuum pump for
continuous suction for about 3.0 minutes, till the filtration is completed.
 Remove the filter paper from the filtration assembly. Dry it in an oven at 103 Co
for 1.0 hour. Cool, desiccate and weight it again (Say B1 mg)

Calculation:
TSS, mg/L = [(B1 – A1)]x1000/ml of sample
Where’
A1 = weight of the filter only (mg)
B1 = weight of the filter + dried residue

(Signatures Laboratory Demonstrator with date)

 7

TDS (Total Dissolved Solids),

 Transfer the filtrate from the filtration assembly to the evaporating dish (china
dish), evaporate to dryness at 180±2oC for 1.0 hour and weight it again (say B2)

Calculation;
TDS, mg/l = [(B2 – A2)]/ml of sample
Where’
A2 = weight of the china dish only (mg)
B2 = weight of the dish + dried residue

Observations/Results:

Sample TDS TSS

1
2
3

Total Fixed Solids (TFS)

 The residue from TDS, TSS, TS, ignite an evaporating dish in a muffle furnace at
550±5 Co for 1.0 hour, Cool, desiccate and weight it again (say A3 mg).

Calculation:
TFS, mg/L = [(B3 – A2)]x1000/ml of sample
Where’
B2 = weight of the dish (mg)
B3 = weight of the dish + dried residue after ignition

TVS (Total Volatile Solids),

 The residue from TDS, TSS, TS Ignite an evaporating dish in a muffle furnace at
550±5 Co for 1.0 hour, Cool, desiccate and weight it again (say A3 mg).

(Signatures Laboratory Demonstrator with date)

 8

Calculation;
TVS, mg/l = [(B3 – B2)]x1000/ml of sample
Where’
B3 = weight of the dish + dried residue after ignition (mg)
A2 = weight of the dish + dried residue

Observations/Results:

Sample TFS TVS

1
2
3

Comments:
__________________________________________________________________________________________________
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(Signatures Laboratory Demonstrator with date)

 9

DETERMINATION OF CHLORIDES IN WATER/WASTEWATER

(Argentometric Method)

Reagents
 Potassium Chromate indicator
 Standard Silver Nitrate Titrant (0.0141N)
 Standard Sodium Chloride (0.0141N)
 Sodium Hydroxide Suspension
 Sodium Hydroxide ( 1.0N) & Sulfuric Acid (1.0N)
 Hydrogen Peroxide

Procedure
Standardization of AgNO3 (0.0141N) with NaCl (0.0141N)

Method
Take a known volume of NaCl (0.0141N) in a flask (say 10ml) add a few drops of
potassium chromate indicator. The color of the NaCl solution will be “yellow”. Titrate
it against AgNO3 (from burette), till the color becomes “brick red”. Take at least three
reading from the burette

Using equation: Silver Nitrate = Sodium Chloride

N1V1 = N2V2
Hence: N1 = [0.0141 * 10ml] * [1/V1]
V1 = The volume of AgNO3 is determine from burette
 Now take a 100ml of the sample.
 Add 1.0ml Potassium Chromate (K2CrO4)
 Titrate it with Silver Nitrate (AgNO3). Till the color of the solution changes form
“yellow” to “brick - red”
 Take at least three readings from the burette. Suppose the mean volume of AgNO3
consumed is “A ml”.

(Signatures Laboratory Demonstrator with date)

 10

 Repeat the same procedure for blank/deionized water/distilled water, instead of

sample. Let the volume of AgNO3 consumed is “B ml”. This step is performed to
calculate the amount of AgNO3 consumed by the indicator (K2CrO4 )
 Using the following expression the conc. of chloride is calculated as;
Cl- mg/L = [(A - B) * N * 35450] / [ ml of sample]
(where, N = Normality of AgNO3)
OR
mg/L NaCl = mg/L Cl- * 1.65 (23 + 35.5 = 58.5)

Observations/Results:

Sample Cl-
1
2
3

Comments:
__________________________________________________________________________________________________
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(Signatures Laboratory Demonstrator with date)

 11

DETERMINATION OF SULFATES USING SPECTROPHOTOMETER

Apparatus/Chemicals: Spectrophotometer with cells, Samples, Distilled water,

Sulfates pillows/reagents, Beaker etc

Procedure:
 Enter the stored number for Sulfates e.g press 680.
 Dial wavelength to 450 nm
 Fill the sample cell with 25 ml of sample.
 Fill another cell with 25 ml deionized water
 Add the contents of one Sulfates Reagent powder pillow to each cell.
 Press SHIFT TIMER.
 When beeps, press SHIFT TIMER and 5 minutes reaction duration will begin.
 When the Timer beeps, place the blank (deionized) sample and close the light
shield.
 Press ZERO buttons, the display will show zeroing ….. And then 0.00 mg/L SO4-2 .
 Place the sample cell and close the light shield.
 Press READ and display will show READING …. And then result will be display.
X mg/L SO4-2

Observations/Results:

Sample X mg/L SO4-2

1
2
3

Comments:
__________________________________________________________________________________________________
__________________________________________________________________________________________________
___________________________________________

(Signatures Laboratory Demonstrator with date)

 12

DETERMINATION OF NITRATE USING SPECTROPHOTOMETER

Apparatus/Chemicals: Spectrophotometer with cells, Samples, Distilled water,

Nitrate pillows/reagents, Beaker etc
Procedure:
 Press SHIFT (release) then TIMER, display will show 1 minute countdown
 Shake the cell vigorously until the timer beeps
 Press SHIFT then TIMER again. Spectrophotometer will begin five minute
reaction period countdown
 While waiting, fill the pour-through funnel with filtered sample, this will be the
blank against which the color of the reacted sample will be compared. The
spectrophotometer reports the difference between these two as the final
composition.
 When the timer beeps, the display will show mg/L NO3-N HR
 Press ZERO (this measures and sets the blank concentration). The display will
show: Zeroing ? Then, 0.0 mg/L NO3-N HR.
 Pour the reacted sample into the pour-through funnel
 wait until the water level in the funnel stops falling
 Press READ The display will show: ``Reading'' then the result in mg/L nitrate
nitrogen (NO3-N) will be displayed.
 Record the value in your notebook.

Observations/Results:

Sample Value
1
2
3
Comments:
__________________________________________________________________________________________________
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_____________________________________________________________________________________________

(Signatures Laboratory Demonstrator with date)

 13

DETERMINATION OF PHOSPHATE USING SPECTROPHOTOMETER

Apparatus/Chemicals: Spectrophotometer with cells, Samples, Distilled water,

Phosphate pillows/reagents, Beaker etc

Procedure:
 Enter the stored number for Phosphorous, reactive e.g press 540.
 Dial wavelength to 420 nm
 Fill 50 ml flask with 25ml of sample.
 Add the contents of one Potassium Persulfate Powder Pillow and swirl to mix.
 Add 2.00 ml of 5.25N Sulfuric Acid solution.
 Place the flask on a hot plate and boil gently for 30 minutes.
 Cool the sample to the room temperature.
 Add 2.0 ml of Sodium Hydroxide solution and swirl to mix.
 Fill the sample cell with 25 ml of sample.
 Fill another cell with 25 ml of Sample (Blank).
 Press SHIFT TIMER. 7 minutes reaction duration will begin.
 When the Timer beeps, place the blank and close the light shield.
 Press ZERO buttons, the display will show Zeroing ….. And then 0.00 mg/L PO43-.
 Place the sample cell and close the light shield.
 Press READ and display will show READING …. And then result will be display.
 X mg/L PO43-

Observations/Results:

Sample I II III
Value

Comments:

__________________________________________________________________________________________________
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___________________________________________

(Signatures Laboratory Demonstrator with date)

 14

DETERMINATION OF HARDNESS BY EDTA

Apparatus/Chemicals:
Burette with stand, Titration flasks, Beakers, Cylinders, D.W, EBT, EDTA Sol, Buffer
Sol, Sodium Hydroxide sol, Hydoxynaphthol blue indicator, Complexing agents (Mg
CDTA) etc

Procedure:
 Take 50 ml of sample in a flask
 Add 2.0ml of buffer solution
 Add 2 drops of EBT indicator
 Note the color of the sample (i.e. wine red)
 Titrate the sample against the standard EDTA (0.01M) till the color changes from
“red” to “blue”
 Note the volume of EDTA consumed from the burette reading (say “A ml”)
 Repeat the same procedure for blank, in order to find out the amount of EDTA
consumed by the buffer solution/indicator etc. Let say the volume consumed is “B
ml”

Calculation:
Net volume of EDTA consumed by the sample = (A - B) ml
Since 1.0 mg CaCO3 = 1.0 ml EDTA sample
Therefore, total Hardness (mg CaCO3/L) = [(A - B) x 1000] / ml of sample

Observations/Results:

Sample I II III
Value

Comments:
__________________________________________________________________________________________________
__________________________________________________________________________________________________
___________________________________________

(Signatures Laboratory Demonstrator with date)

 15

DETERMINATION OF DISSOLVED OXYGEN (DO METER)

Apparatus: DO Meter, Beaker, Water/Wastewater Sample

Procedure:
 Take a beaker and sterilize it and then put sample of water in it.
 Clean the probe of DO meter and then dip it into the beaker.
 Place the Beaker on Magnetic Mixer to ensure that water flows through the probe
sensor.
 Press On button and then Contd… will be displayed.
 Then press Read button and after a while result will be displayed in X ppm.
 ppm = mg/L, so the displayed result will be DO Xmg/L

Observations/Results:

Sample I II III
Value

Comments:
__________________________________________________________________________________________________
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___________________________________________

(Signatures Laboratory Demonstrator with date)

 16

DETERMINATION OF BIOCHEMICAL OXYGEN DEMAND

(DILUTION METHOD)

Apparatus/Chemicals:
DO meter, flasks, BOD bottles, incubator, dilution water, magnesium sulfate solution,
calcium chloride solution, ferric chloride solution, acid and alkali solutions,
phosphate buffer solution

Measurement
Principle:
The test measures the oxygen utilized during specific period of incubation for the
biochemical degradation of organic material, by computing the difference between
initial and final DO.
Sampling and Storage
Samples for BOD analysis may degrade significantly during storage between
collection and analysis, resulting in low BOD values.
Samples should be analyzed immediately after collection, if not they should be kept
near freezing temperature (oC)
Seeding
It is necessary to have a population of microorganisms capable of oxidizing the
biodegradable organic matter in sample. Domestic wastewater, unchlorinated
effluents and surface water receiving wastewater discharges contain satisfactory
microbial population and may not be seeded. But untreated industrial effluent,
disinfected water/wastes, effluent with high temperature or pH value, lack
significant microbial. For such sample add seeds (mixed group of microorganisms).
Raw sewage or supernatant portion of the domestic sewage, after settling at 20 oC for
at least 1.0 hour, is preferred seed.
Temperature
The oxidative reactions involved in the BOD test are temperature dependent.
Temperature effects are held constant by performing the test at 20 oC, which is more
or less, a medium value as far as natural bodies of water are concerned. Since water
at 20 oC has limited dissolved oxygen approximately 9mg/L, it is necessary to dilute
the samples for BOD test to ensure the presence of excess dissolved oxygen

(Signatures Laboratory Demonstrator with date)

 17

Preparation of Dilution Water

 Take appropriate amount of distilled water in a suitable bottle.
 Add phosphate buffer solution (for pH adjustment at about 7.0)
 Add the following nutrients as 1mL/1L of distilled water,
- Magnesium Sulfates
- Calcium Chloride
- Ferric Chloride
 Aerating sample with compressed filtered air/ or manually by mixing
- Stored in an air sealed bottle (at 20oC)
 The DO uptake of dilution water in 05 days at 20oC should not be more than
0.2mg/L
Sample Preparation
 Neutralized the caustic alkalinity or acidity, using phosphate buffer solution
 Avoid sample containing residual chlorine by sampling a head of chlorination
 If the sample has been chlorinated but no detectable chlorine residual is present,
seed the dilution water
 Or determine the amount of Sodium thiosulfate
 to add to the sample
 To eliminate the effects of phenol, heavy metals or cyanide etc. dilute with high
quality and amount of water.
 For cold samples, first allow temperature to reach 20oC before testing

Reagents:
 Dilution Water
 Magnesium Sulfate Solution
 Calcium Chloride Solution
 Ferric Chloride Solution
 Acid and Alkali Solutions
 Phosphate Buffer Solution

Procedure:
 Take five clean and sterilized BOD bottles of 300mL capacity.

(Signatures Laboratory Demonstrator with date)

 18

 Use four BOD bottles for the preparation of different dilutions of a sample and
one bottle for blank (dilution water)
 Take 4 different volumes of the sample around the volume limits, given in table.
Fill each bottle with dilution water up to 300mL.
 Measure initial DO, D0 of each of the sample dilutions and of the blank with DO
meter
 Place the stoppers on each bottle in such way that no air bubbles remains inside
the bottles.
 Incubate all the bottles at 20 oC for five days
 Measure final DO, D5 of each of the sample dilutions and of the blank after
incubation period.
 Select those dilutions, which show drops of DO within the limits given below;
 It is desirable to have at least 2mg/L DO left unused after five-day incubation.
Depletion of 50% DO should be left after five day incubation is most desirable.
 e.g. 4.5mg/L of DO left out of initial DO of 9.0mg/L.

(Signatures Laboratory Demonstrator with date)

 19

Calculations:
BOD5 mg/L = (D0 – D5) x Dilution Factor (D.F)
Where,
D0 = DO conc. at Zero/Initial Day
D5 = DO conc. at 5th Day
D.F = Dilution Factor [300mL/mL of sample]
e.g. if we add 5mL of sample in 300mL of dilution water than,
D.F = 300mL/05mL = 60

1 2 2 2 2
DO0
DO5

Comments:
__________________________________________________________________________________________________
__________________________________________________________________________________________________
___________________________________________

(Signatures Laboratory Demonstrator with date)

 20

DETERMINATION OF CHEMICAL OXYGEN DEMAND (COD)

(Open Reflux Method)

Apparatus:
Blender, DRB200 Reactor with 13-mm wells (use adapters with 16-mm holes), 1-
COD TNTplus™ vials for the appropriate concentration range, Pipette for 2.0 mL
Sample, Test Tube Rack

Procedure:
 Turn on the DRB200 Reactor. Preheat to 150°C. For DRB200 Reactors with 16-
mm wells, insert a 16- mm to 13-mm adapter sleeve into each well before turning
on the reactor.
 Homogenize 100 mL of sample for 30 seconds in a blender. For samples
containing large amounts of solids, increase the homogenization time.
 To help to make sure that a representative portion of sample is analyzed, pour the
homogenized sample into a 250-mL beaker and gently stir with a magnetic stir
plate.
 Carefully pipet 2.0 mL of sample into the vial. Cap and clean the outside of the
vial.
 Hold the vial by the cap over a sink. Invert gently several times to mix. The sample
vials will become very hot during mixing. Place the vial in the preheated DRB200
Reactor. Close the protective lid.
 Heat for two hours.
 Turn the reactor off. Wait about 20 minutes for the vial to cool to 120°C or less.
 Invert the vial several times while still hot.
 Place the vial into a rack to cool to room temperature.
 Thoroughly clean the outside of the vial.
 Enter the stored program number 435 and then set wavelength at 620 nm.
 Then the spectrophotometer will display Zero Sample then mg/L COD HR.
 Insert the Blank into the cell holder with the marker to the right. Close the lid.
 Press Zero and it will display 0 mg/L COD HR.

(Signatures Laboratory Demonstrator with date)

 21

 Insert the sample vial into the cell holder with the marker to the right. Close the
lid.
 Press READ. The display will show
 Reading ….. Then the result will be displayed.
X mg/L COD.

Observations/Results:

Sample mg/L COD

1
2
3

Comments:
__________________________________________________________________________________________________
__________________________________________________________________________________________________
___________________________________________

(Signatures Laboratory Demonstrator with date)

 22

DETERMINATION OF TOTAL COLIFORM

(Membrane Filtration Method)

Apparatus/Chemicals:
Petridish, membrane filter (0.45microns), vacuum pump apparatus, incubator,
forceps, FC-Agar medium

Procedure:
 Place sterile petridish on the table and remove the lid.
 Carefully pour FC-Agar medium in the dish
 Set up the membrane filter assembly
 Place membrane filter in the assembly using a sterile forceps
 Pour 100mL of sample into the funnel
 Apply vacuum and filter the sample
 Turn off the vacuum
 Using sterilized forceps, transfer the filter immediately to the previously
prepared petridish.
 Invert the petridish lid and place it in the incubator at 41.5oC for 18 hours.
 Remove the petridish after 18 hours from the incubator and count the colonies
with that are yellow in color using a magnifier or a low-power microscope.

(Signatures Laboratory Demonstrator with date)

 23

Counting:
Fecal coliform colonies are yellow in color. This color is produced due to lactose
fermentation (confirms fecal coliform colonies).

Total Coliform colonies /100 mL = Coliform colonies counted x 100

mL sample used

Observations/Results:

Sample No. of Total Coliform

1
2
3

Comments:
__________________________________________________________________________________________________
__________________________________________________________________________________________________
___________________________________________

(Signatures Laboratory Demonstrator with date)