EnvEng 6052 / ENVIRONMENTAL ENGINEERING

LABORTORY (PG – MSc)

ACADEMIC YEAR: 2019-2020
Year: 1st and Semester: 2nd

Name:_________________________________

Id No:__________________________________

Environmental Engineering Chair

Department of Water Supply and Environmental Engineering
Faculty of Civil and Environmental Engineering
Jimma Institute of Technology, Jimma University,
Jimma, Ethiopia

Submitted to: Dr. P. Asaithambi [Link]., Ph.D., Date of Submission:______________.

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 1

 EnvEng 6052 / ENVIRONMENTAL ENGINEERING
LABORTORY (PG – MSc)
Environmental Engineering Chair
Department of Water Supply and Environmental Engineering
Faculty of Civil and Environmental Engineering
Jimma Institute of Technology, Jimma University.
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Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 2

Exp. No: Date:
Determination of Color
Aim: To determine the color of a given water sample using visual comparison methods.

Introduction:
 Coloured water is not acceptable for drinking (Aesthetic as well as toxicity reasons).
 Industrial wastewater requires colour removal before discharge into water bodies.
 The term colour means true colour that is the colour of water.

Visual Comparison Method

 Colour of the sample is determined by visual comparison with known concentration of
coloured solutions prepared by diluting stock platinum cobalt solution OR properly
calibrated glass coloured disk is used for comparison.
 This method is useful for potable water and water in which colour is due to naturally
occuring materials.
 This method is not applicable to most highly coloured industrial wastewater.

Materials Required:

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 3

Apparatus
(i). Beaker, etc.,
Reagents
(i) Water samples, etc.,

Procedure:
Determining the colour in the field is relatively easy. Pour an aliquot of approx.10mL of sample
into a glass test tube and judge the colour observed. Consider one of the following options:
Light brown, Brown, Dark brown, Light green, Green, Dark green, Clear, Other specify.

Results:

S/No Type of sample Colour of the sample

1. Industrial wastewater
2. Tap wastewater
3. Surface water
4. Wastewater
Environmental Significance:
(i), etc.,

Exp. No: Date:

Determination of Conductivity
Aim: To determine the conductivity value of a given water sample using conductivity meter.
Introduction:
 EC is reciprocal of electrical resistance (Ohms); it indicates presence of cations (K +, Na+,
Ca2+….) and anions (Cl-, HCO3-, SO42- ) in water.
 It is used to measure the concentration of dissolved solids which have been ionized in a
polar solution such as water.
 Unit: μS/cm or mS/cm (S=Siemens). Values in most natural waters: 10-1000 μS/cm and salt
water: mS/cm.
 EC is often used as (fully automated) water quality check for TDS (total dissolved solids),
e.g. for drinking water.
 In agriculture for quality of the irrigation/drainage water.
 Measuring the conductivity is an accurate way to determine the salinity.
Materials Required:
(i). Apparatus – conductivity meter with measuring cell, water sample, beaker, etc.,
(ii). Chemicals – KCl 0.1N, NaCl, etc.,
Procedure:

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 4

1. Switch on the conductivity meter.
2. Prepare KCl (0.1N) solution.
3. Calibrate the conductivity meter using the standard solution of 0.1N KCl.
4. Read the conductivity meter by inserting the sample.
5. Adjust the conductivity using NaCl or KCl solution.
Tabulation

Results: The conductivity value of given sample is: - μS/cm or mS/m

Environmental Significance:
1. It is useful to assess the source of pollution.
2. Electrical conductivity measurements are often employed to monitor desalination plants.
3. etc.,

Exp. No: Date:

Determination of pH
Aim: To determine the pH value of a given sample using pH meter.
Introduction:
• pH= -log[logH+]
• pH is a physical unit used to measure the degree of acidity (or) alkalinity of a solution.
• Measure ranging from 0 to 14.
• Value is directly related to the H+/OH-
• pH = 0 solution become more acidic H+ > OH-
• pH = 14 solution become more alkaline OH- > H+
• pH = OH- = H+ solution is neutral.
• If equal amount of H+ and OH- ion are present, the solution is neutral and is characterized by
a pH of value is 7.
[Link] Type of sample Temperature, Conductivity,
⁰C (µS/cm)
1. Surface water
2. Wastewater
3. Industrial wastewater
4. Tap wastewater
Materials Required:
(i). Apparatus – pH meter, water sample, beaker, etc.,
(ii). Chemicals – H2SO4, NaOH solution, etc.,
Procedure:
1. Switch on the pH meter.

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 5

2. Prepare buffer solution of 4.0. 7.0 and 9.2.
3. Calibrate the pH meter using buffer solution and adjust the calibration knob.
4. Read the pH meter by inserting the sample.
5. By adjust the pH using 0.1M solution of NaOH and H2SO4.
Tabulation:
[Link] Type of sample Temperature, ⁰C pH
1. Industrial wastewater
2. Tap water
3. Surface water
4. Wastewater

Results: The pH value of given water sample is:_________.

Environmental Significance:
1. etc.,

Exp. No: Date:

Determination of Turbidity
Aim: To determine the turbidity of the a given sample

Introduction:

Materials Required:
(i). Chemicals – Hydrazine sulphate, Hexa methylene tatramine
(ii). Apparatus – Nephelometric turbidity meter, Sample tubes

Procedure:
1. Switch on the turbidity meter.
2. Prepare 400 NTU solution.
3. Calibrate the turbidity meter to 400 NTU using the standard solution by adjust the calibration
knob.
4. Calibrate the turbidity meter to 0 NTU using the distilled water by adjust the calibration knob.
5. Read the turbidity meter by inserting the sample.

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 6

Tabulation
S.N Type of sample Temperature, ⁰C Turbidity, (NTU)
o
1. Surface water
2. Wastewater
3. Industrial wastewater
4. Tap water

Results: The turbidity of the given sample= ----------------------NTU

Environmental Significance:
• Clarity of water is important for human consumption.
• Manufacturing processes such as beverages, and many food products need clear water.

Exp. No: Date:

Determination of Total Solids

Aim: To determine the total solids present in the given sample.

Introduction:
• Solids – Suspended or dissolved solids.
• Physically isolated either through filtration, evaporation or combustion.
• Solids – (i) Filterable – (a) Settleable and (b)non-settleable
(ii) non filterable
• Solids –(i) Organic (Carbon, hydrogen, nitrogen, oxygen, etc.,)
(ii) In-organic (Salts, metals, etc.,)
• Total solids – the material residue left in the vessel after evaporation of sample and its
subsequent drying in an oven at defined temperature.
• Measurement of solids can be made in different water samples (Industrial, domestic and
drinking water, etc.,.)
• Total solids = TDS+ TSS.

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 7

Materials Required:
(i). Apparatus – Evaporating dish, oven, desiccators, weighing balance.

Procedure:
1. Wash and wipe the evaporating dish(Porcelain dish) and dry it in a hot air oven for dryness.
2. Measure the initial weights of dishes by using electronic balance(W1, (g)).
3. Take 10 mL of unfiltered sample in a china dish and evaporate in oven at 103⁰C to 110⁰C.
4. Cool the container to dryness in desiccators and weigh the dishes again.
5. Note the increase in weight(W2, (g))

Calculation
(W 2 −W 1 )∗1000
∗1000
Amount of total solids present in the water sample = Volumeofsampletaken (mL)

Results: Amount of total solids present in the sample is = ________ mg/L

Environmental Significance:
1. Total solids determination is used to assess the suitability of potential supply of water for
various uses. In cases, in which waster softening is needed, the types of softening procedure
used may be dictated by the total solid content.
Exp. No: Date:

Determination of Alkalinity
Aim: To estimate the alkalinity present in the given water sample.

Introduction:
• Alkalinity measures the acid-neutralizing capacity of a water sample.
• The alkalinity in a water sample measured by the amount of standard acid needed to lower the
pH.
• Based on stoichiometry of the reaction and number of moles of Sulphuric(H 2SO4) acid needed
to reach the end point, the concentration of alkalinity in water is calculated.

Materials Required:
Apparatus required
Burette, Pipette, Conical flask, Measuring Jar, Standard flask, Beakers, etc.,
Chemicals reagent required
1. Standard sulphuric acid (0.02N),
2. Phenolphthalein indicator,

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 8

3. Mixed Indicator.
Procedure:
Observation
Titration 1 (For Phenolphthalein Alkalinity)
Burette solution: H2SO4
Pipette solution: Sample
Indicator: Phenolphthalein
End point: Disappearance of pink color
Types of water: Tap

S.N Volume of sample in mL Burette readings Volume of H2SO4, used mL

Initial, mL Final, mL
o
1.
2.
3.

Titration 2 (For Total Alkalinity)

Burette solution: H2SO4
Pipette solution: Sample
Indicator: Mixed
End point: Red color
Types of water: Tap

S.N Volume of sample in mL Burette readings Volume of H2SO4, used mL

Initial, mL Final, mL
o
1.
2.
3.

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 9

Results:
Total alkalinity present in the given sample = --- mg/L
Phenolphthalein Alkalinity = --- mg/L

Environmental Significance:
1. Chemical coagulation of water and wastewater
2. Water softening
3. Corrosion control
4. Effluent of wastewater

Exp. No: Date:

Determination of Acidity
Aim: To estimate the acidity present in the given water sample.

Introduction:
 Acidity is a measure of the capacity of water to neutralise bases.
 Acidity is the sum of all titrable acid present in the water sample. Strong mineral acids,
weak acids such as carbonic acid, acetic acid present in the water sample contributes to
acidity of the water.
 Usually dissolved carbon dioxide (CO2) is the major acidic component present in the
unpolluted surface waters.

Materials Required:
Apparatus required
1. Burette with Burette stand

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 10

 2. porcelain tile
3. 500 mL conical flask
4. Pipette with elongated tips
5. Pipette bulb
6. Conical flask
7. Measuring cylinders
8. Wash Bottle and Beakers

Chemicals required

1. Sodium Hydroxide
2. Phenolphthalein
3. Methyl Orange
4. Ethyl alcohol
5. Distilled Water

Procedure:
Observation
Burette solution: NaOH (0.02N)
Pipette solution: Sample
Indicator: Methyl Orange
End point: Yellow Color
Indicator: Phenolphthalein
End point: Appearance of pink color

Types of water: Tap

[Link] Volume of Methyl Orange Indicator Phenolphthalein

sample in ml Initial, mL Final, NaOH Indicator
mL Used V1 Initial, Final, NaOH
mL mL Used V2

1.
2.
3.
4.

Calculation:

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 11

Mineral Acidity due to mineral acids as mg/L of CaCO 3 = V1*N*50*1000 / Volume of sample
taken
Phenolpthalein Acidity due to mineral acids as mg/L of CaCO 3 = V2*N*50*1000 / Volume of
sample taken
Total Acidity as mg/L of CaCO3 = Mineral Acidity + Phenolpthalein Acidity

Results:
Methyl orange acidity : ------- mg/L
Phenolpthalein Acidity : ------- mg/L
Total Acidity : ------- mg/L

Environmental Significance:
 Acidity interferes in the treatment of water. Carbon dioxide is of important considerations in
determining whether removal by aeration or simple neutralisation with lime /lime soda ash
or NaOH will be chosen as the water treatment method.
 The size of the equipment, chemical requirements, storage spaces and cost of the treatment
all depends on the carbon dioxide present.
 Aquatic life is affected by high water acidity. The organisms present are prone to death with
low pH of water.
 High acidity water is not used for construction purposes. Especially in reinforced concrete
construction due to the corrosive nature of high acidity water.
 Water containing mineral acidity is not fit for drinking purposes.
 Industrial wastewaters containing high mineral acidity is must be neutralized before they are
subjected to biological treatment or direct discharge to water sources.

Exp. No: Date:

Determination of Hardness
Aim: To determine the total hardness of the given sample.

Introduction:
• Generally hardness of water is defined as the measure of capacity of water to precipitate
soap i.e., the capacity of the water to form lather with soap.
• Hard water contains dissolved minerals such as Ca2+, Mg2+, Fe3+, SO42- ,etc.,
• The degree of hardness is measured in Parts Per Million(ppm) or Grams per Gallon(GPG).
• Water with different degrees of hardness is used for different purposes such as household
and industrial purposes.

Materials Required:

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 12

Apparatus - Burette with Burette stand, porcelain tile, 500 mL conical flask, Pipette with
elongated tips, Pipette bulb, Conical flask, Measuring cylinders, Wash Bottle and Beakers

Reagents
1. Buffer (pH 10): mixture of ammonia and ammonium chloride solutions in water.
2. EDTA (Disodium salt of EDTA) for precise end point.
3. Erichrome Black T.

Procedure:
Observation
Burette solution: EDTA
Pipette solution: Sample
Indicator: EBT
End point: Wine red to steel blue

[Link] Volume of Burette readings Volume of EDTA mL

sample in mL Initial, mL Final, mL

1.
2.
3.
4.
Calculation

Amount of hardness present

in the given sample = (Volume of EDTA*1000) / ( Volume of sample taken)

Total hardness = Temporary hardness + Permanent hardness

Temporary hardness = Total hardness – Permanent hardness

Results: The amount of Total hardness present in the given sample = -- mg/L .

Environmental Significance:
 It is essential to know the hardness of the water since it defines the purpose of it.

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 13

 Hard water is not a health hazard but it cannot be used in industrial applications such as in
boilers, pipes, etc.,
 It has the tendency of forming CaCO3 precipitate which later results in sludge/scale formation.

Exp. No: Date:

Determination of Chloride

Aim: To estimate the amount of chlorides present in the given water sample.

Introduction:
 Chloride in the form of Chloride (Cl−) ion is one of the major inorganic anions in water and
wastewater.

Materials Required:
Apparatus
Pipette, burettes, conical flasks, Volumetric flask, 1𝐿 Volumetric flasks, wateman filter
paper, Measuring equipments, distilled water, Bunsen burner.
Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 14
Reagents
1. Silver nitrate (0.014N)
2. Sodium Chloride (0.014N)
3. Potassium chromate indicator

Procedure:
1. 20 mL of sample from wastewater.
2. Adjust the pH to be between 7 & 8 by using H2SO4 (or) NaOH with pH meter.
3. Add 1 mL of potassium chromate to light yellow color.
4. Titrate with std AgNO3 solution till the colour change from yellow to brick red.
5. Note the volume of AgNO3 solution.
6. For better accuracy titrate distilled water in the same manner.
7. Volume of AgNO3 added for distilled water.

Calculation

Chloride (mg/L) = (A-B) x N x 35.45 x 1000

mL sample
Where A = mL AgNO3 required for sample, B = mL AgNO3 required for blank N = Normality of
AgNO3 used.

Results: Amount of chlorides present in the given sample = -- mg/L

Environmental Significance:
 The chloride concentration is higher in wastewater than in raw water because of sodium
chloride.
 Average consumption 6 g of chlorides/person/day.
 Measured chlorides ions can be used to know salinity of different water sources.
 It is also interferes the COD.
 It can also corrode concrete.
 Chloride determination natural water are useful in the selection of water supplies for human
use.
 It is used to determine the type of desalting apparatus to be used.
 It is used to control pumping of ground water from location where intrusion of seawater is a
problem.

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 15

Exp. No: Date:
Determination of Dissolved Oxygen
Aim: To determine the dissolved oxygen value of a given sample using Electrometric method.

Introduction:

Materials Required:
(i). Apparatus – Battery-powered meter, Oxygen-sensitive membrane electrode, water sample,
beaker, etc.,

(ii). Chemicals – Na2SO3, CoCl2, etc.,

Procedure:

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1. Follow exactly the calibration procedure described in the manufacturer’s operating
instructions. Generally, electrodes are calibrated by reading against air or against a sample of
known dissolved oxygen content. This “known” sample could be one for which dissolved
oxygen concentration has been determined by the Winkler method or one that has been saturated
with oxygen by bubbling air through it. The zero end of a calibration curve can be determined by
reading against a sample containing no dissolved oxygen, prepared by adding excess sodium
sulphite, Na2SO3, and a trace of cobalt chloride, CoCl2, to the sample.
2. Rinse the electrode in a portion of the sample which is to be analysed for dissolved oxygen.
3. Immerse the electrode in the water, ensuring a continuous flow of water past the membrane to
obtain a steady response on the meter.
4. Record the meter reading and the temperature, and the make and model of the meter.
5. Switch the meter off and pack it and the electrode in the carrying case for transport.
Tabulation:
[Link] Type of sample Temperature, ⁰C DO, mg/L
1. Surface water
2. Wastewater
3. Industrial wastewater
4. Tap water

Results: The dissolved oxygen value of given water sample is :

Environmental Significance:
2. etc.,

Exp. No: Date:

Determination of Chlorine
Aim: To determine the available chlorine in the given sample.

Introduction:
• Chlorine is one of the most widely used disinfectants.
• It is very applicable and very effective for the deactivation of pathogenic microorganisms.
• Chlorine can be easily applied, measures and controlled.
• It is relatively cheap.
• Chlorine has played an important role in lengthening the life-expectancy of humans.

Materials Required:
Apparatus

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• Burette & Burette Stand
• Porcelain Tile
• Pipettes with elongated tips
• Pipette Bulb
• Wash Bottle
• 250 mL Graduated Cylinder
• 500 mL Conical Flask (Erlenmeyer flask)

Reagents
 Acetic Acid, Conc. (glacial)
 Potassium Iodide, KI, crystals
 Sodium thiosulphate (0.025N)
 Starch indicator
 Distilled or Deionized Water

Procedure:
Observation and Table
Burette Solution: Sodium Thiosulphate
Pipette Solution: Sample
Indicator: Starch
End point: Disappearance of blue color

Burette Reading Volume of

Sample No. Temperature of Volume of (mL) Titrant Residual
Sample Sample (mL) Chlorine
(mL) (Na2S2O3 (mg/L)
(ºC) Initial Final
solution
used)

Calculation

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 18

Results:
The amount of residual chlorine in the given sample = -- mg/L

Environmental Significance:
1.
2.
3.
4. etc.,

Exp. No: Date:

Determination of Determination of Chemical Oxygen Demand (COD)

Aim: To find out Chemical Oxygen Demand of a given waste water sample.

Introduction:
 The organic matter present in sewage can be measured in number of ways.
 Organic matter is often assessed in terms of O 2 required to completely oxidize the organic
matter to CO2, H2O and other oxidized species.
 The O2 required to oxidize the organic matter present in given waste water can be
theoretically calculated.

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 19

 If the organic compounds and their concentration are known the oxygen demand of the
sample can be accurately computed but it is impossible to know the details of organic
compounds present in raw water or waste water.
 The COD is therefore determined by performing a laboratory test on given water sample
using a strong oxidant like dichromate solution.
 It is expressed in mg/L also referred to as ppm, which indicates the mass of O 2 consumed per
liter of solution.

Apparatus Required:
COD Digester, Burette & Burette stand, COD Vials with stand, 250 mL conical flask
(Erlenmeyer Flask), Pipettes, Pipette bulb, Tissue papers, Wash Bottle, etc.,

Chemicals required
Potassium dichromate, Sulfuric acid, Ferrous ammonium sulphate, Silver sulphate, Mercury
sulphate, Ferroin indicator, Organic free distilled water.

Preparation of reagents
1. Standard K2Cr2O7 Reagent - Digestion Solution
a. Weigh accurately 4.913g of potassium dichromate, previously dried at 103ºC for 2 - 4 hrs
and transfer it to a beaker.
b. Weigh exactly 33.3g of mercuric sulphate and add to the same beaker. Measure accurately
167 mL of concentrated sulphuric acid using clean dry measuring cylinder and transfer it
to the beaker. Dissolve the contents and cool to room temperature. (If not dissolved keep
it over night).
c. Carefully transfer the contents to the 1000 mL standard flask and make up to 1000 mL
using distilled water.
This is the standard potassium dichromate solution to be used for digestion.
2. H2SO4 Reagent - Catalyst Solution
Weigh accurately 5.5 g silver sulphate crystals to a dry clean 1000 mL beaker. To this carefully
add about 500 mL of concentrated sulphuric acid and allow standing for 24 hours (so that the
silver sulphate crystals dissolve completely).
3. Standard FAS solution
Weigh accurately 39.2g of ferrous ammonium sulphate crystals and dissolve it in distilled water.
Carefully transfer the contents to the 1000 mL standard flask and make up to 1000 mL mark
using distilled water.

Procedure
1. Wash 25 mL of COD vials.

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 20

 2. In COD vials put 0.05 g of HgSO 4 + 2.5 mL sample + 1.5mL K 2Cr2O7 + 3.5 mL concentrated
H2SO4.
3. Add small amount of silver sulphate.
4. Shake well and reflux for 2 hr.
5. Cool and add little amount of distilled water to the flask through the condenser.
6. Titrate the solution in the flask against FAS using Ferroin indicator
7. End point green color to reddish brown
NOTE: For blank, add 2.5 mL distilled water instead of sample. Rest of the procedure is the
same.

Tabulation
Burette solution: FAS
Pipette solution: Given Sample + Reagent
Indicator: Ferroin Indicator
End point: Blue green to Reddish Brown

[Link] Volume of Burette readings Volume of FAS,

sample in mL Initial, Final, mL mL
mL

1.
2.
3.
4.

Calculations
The COD in mg/L is determined by the formula,

(A - B) x N x 8000 (1)
COD, mg/L=
mL sample taken

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 21

Where, A = mL of FAS required for blank solution, B = mL of FAS requires for sample solution
and N = Normality of FAS.

Results: Chemical Oxygen Demand of a given water sample = ------------mg/L

Environmental Significance:
 The COD test is used extensively in the analysis of industrial wastewater.
 It is particularly valuable in surveys designed to determine and control losses to sewer
systems.
 The test widely used in the place of BOD in the operation of treatment facilities because of
the speed with which the results can be obtained.
 It is useful to assess strength of wastes, which contain toxins and biologically resistant
organic substances.
 The ratio of BOD/COD is useful to assess the amenability of waste for biological treatment.
Ratio of BOD/COD > than or = to 0.8 indicates that wastewater highly amenable to the
biological treatment.

Exp. No: Date:

Determination of Oil and Grease
Aim: The aim of experiment is to determine oil and grease in water, wastewater using Partition-
Gravimetric method.

Introduction
 Oil and grease is an important parameter for water quality and safety.
 The presence of oil and grease always can be seen in a water sample.

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 Regulatory bodies worldwide set limits in order to control the amount oil and grease entering
the water reservoirs or the sea through industrial discharges and also limit the amount
present in drinking water.
 The term grease is used to describe semisolid lubricants.
 Oil and grease in the water sample are referred to the all the filterable solvent soluble
substances such as elemental sulfur, complex aromatic compounds, hydrocarbon
derivatives of chlorine, sulfur and nitrogen and certain inorganic dyes that are extracted and
recovered.
 Oil and grease is any material recovered as a substance soluble in petroleum ether, Hexane or
trichlorotrifluroethane.
 Oil and grease have low solubility and tend to form floating scum layers in surface waters
and waste waters.
 Oil and grease originate from domestic and industrial wastes, and the main concern is that
they can cause problems in sewage and waste water treatment plant.
 If it is present in excessive amount, they may interfere with aerobic and anaerobic biological
processes and lead to decreased wate water treatment efficiency.
 Oils, fats, waxes, hydrocarbons and high molecular fatty acids contribute to the oil and
grease measurement.

Materials and chemicals required

Petroleum ether, Sulphuric acid (1:2), Ethyl alcohol, Separating funnel(500 mL), Filter paper,
Filter funnel, 200 mL glass beaker, Hot water bath, Electronic balance.

Procedure:
1. Add 10 mL of sulphuric acid, 50 mL of petroleum ether and approximately 3 mL of ethyl
alcohol to 250 mL water sample in a separating funnel and shake well.
2. Allow to stand untill the two layers become distinct.
3. Discard the lower water layer from the funnel and drain. The petroleum ether through a filter
paper inside a filter funnel into the preweighed beaker.
4. Pour some fresh petroleum ether through the filter paper to ensure that no oil and grease
remain stuck to the paper.
5. Place the beaker in the hot water bath and allow the petroleum ether to evaporate.
6. Allow the beaker to cool and weigh.
Calculation

Oil and grease = 1000 × (A-B)/V, (mg/L) (1)

Where, A - Weight of the beaker and residue after evaporation of ether (mg), B - Weight of the
preweighed beaker (mg), V- Volume of the sample (mL).

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 23

Results: The oil and grease of the given wastewater sample = ----------mg/L.

Environmental Significance
 Control of anaerobic waste treatment process.
 Accumulation should be controlled or causes disastrous effect
 Inhibitions caused results in decreased the rate of destruction of volatile.
 Sludge must be removed or transferred.
 Important in detecting presence of unbalanced conditions in anaerobic treatment units.

Exp. No: Date:

Determination of Nitrogen
Aim:- To determine the nitrogen from given sample by using Kjeldahl method.

Introduction
 Total Nitrogen – inorganic nitrogen plus organic nitrogen.
 Total Nitrogen – particulate nitrogen plus dissolved nitrogen

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 24

 Particulate Nitrogen - A significant amount of nitrogen (~20%) is bound to insoluble
organic matter. This nitrogen can eventually oxidize to inorganic nitrogen becoming
bioavailable.
 Dissolved Nitrogen – dissolved organic nitrogen plus dissolved inorganic nitrogen.
 Dissolved Organic Nitrogen – soluble organic compounds containing nitrogen, examples are
humic acids, urea, glycine, nicotinic acid, etc.
 Dissolved Inorganic Nitrogen = Total Inorganic Nitrogen
 Total Inorganic Nitrogen – ammonium ion(NH4+), nitrate ion(NO2-), and nitrite ion(NO3-).
These ions are freely water soluble.

Material and equipment:-

 500 ml Kjeldahl flasks.
 250 ml Erlenmeyer flasks.
 Glass beads.
Chemicals Required:-
 Mercuric oxide, reagent grade.
 Potassium sulphate or anhydrous sodium sulphate, reagent grade.
 Sulphuric acid (98%), nitrogen free.
 Paraffin wax.
 40% solution of NaOH; dissolve 400 g of NaOH in water and dilute to 1,000 mL.
 4% sodium sulphate solution.
 Boric acid indicator solution; add 5 mL of a solution with 0.1% methyl red and 0.2%
bromocresyl green to a saturated boric acid solution.
 Standard solution of 0.1N chlorhydric acid.

Procedure
1) Sample: 5.0 mL fresh cow’s milk in a digestion tube.
2) Reagents for digestion: to each milk sample and also to an empty digestion tube (blank) add
the followings:
- 2 tablets of K2SO4 + Se catalyst
- 10.0 mL of concentrated H2SO4 (98%)
3) Digestion: heat for ca. 30 minutes at 420ºC.
ATTENTION: Do not inhale the gases evolve in reaction 1.
4) Cooling and diluting: let the digestion tubes to cool to 50-60 ºC and add to each add 50 mL of
distilled water.
ATTENTION: Let the tubes stand in air to cool, cold water may break the tubes.
5) Distillation:
i. Place the digested samples in digestion tubes to the distilling unit and add 50.0 mL of 35%
(w/v) NaOH.

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 25

ii. The sample is distilled until 100 mL of distillate are collected in 25.0 mL of 4.0 % (w/v) boric
acid.
6) Titration: add 2-3 drops indicator to the Erlenmeyer flask and titrate it with 0.1 M HCl.
7) Calculate the amount of protein (% protein) and compare the result with the value give on the
milk.
Calculation
Let, Mass of the organic compound = W g
Volume of the standard acid required for complete neutralization of the evolved ammonia = V
mL
Normality of the standard solution of acid = N
From the law of equivalence (normality equation),
1000 mL of 1 N acid = 1000 mL of 1 N NH 3 = 17g(molar mass) NH3 = 14 g nitrogen(Atomic
mass)
Then,
V mL of N acid = V mL of NH3
NV milli equivalent of acid = NV milli equivalent of ammonia

(1)

Then,

(2)

1.4 X normality of H2SO4 x Volume of H2SO4

% of nitrogen in the sample = used for complete neutralization of NH3
Mass of the compound taken
Results: The nitrogen of the given wastewater sample = ----------mg/L.
Environmental Significance:
Exp. No: Date:
Determination of Sulfate
Aim:- Determine sulfate ion concentration in a water sample.

Introduction
 Sulfate are found in appreciable quantity in all natural waters, particularly high in arid and
semi arid regions where natural waters in general have high salt content.

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 26

 Sulfate salts are mostly soluble in water and impart hardness. Water with high concentrations
has a bitter test.
 Sulfate may cause intestinal disorders. These ions can produce hydrogen sulfides as per
following equation (1):

 The sulfate data is used in determining applicability of different water types for their
public and industrial applications.
 It indirectly indicates extent of problems that can arise due to reduction of sulfates to
hydrogen sulfides.
 In addition, sulfate content of organic matter fed to anaerobic digester is important
information as it gives idea of generation of hydrogen sulfides, which needs to be removed.

Material Required:-
Whatman No. 1 filter paper; UV/Vis Spectrophotometer; Magnetic stirrer.
Chemicals Required:-
1. Buffer Solution A: Dissolve 30 g magnesium chloride (MgCl 2.6H2O), 5 g sodium acetate
(CH3COONa.3H2O), 1.0 g potassium nitrate (KNO3), and 20 mL acetic acid (CH3COOH; 99%)
in 500 mL distilled water and make up to 1000 mL.
2. Buffer Solution B (required when the sample SO42-<10 mg/L): Dissolve 30 g magnesium
chloride, 5 g sodium acetate, 0.111 g sodium sulfate, and 20 mL acetic acid (99%) in 500 mL
distilled water and make up to 1000 mL.
3. Dry Barium Chloride (BaCl2) crystals
4. Standard Sulfate Solution: Dissolve 0.1479 g of anhydrous sodium sulfate in distilled water to
make the volume 1 L. This solution contains 100 mg sulfate/L (i.e., 1 mL=100μg SO4 2-). Prepare
standards of various strengths (preferably from 0.0 to 40.0mg/L at the intervals of 5 mg/L by
diluting this stock solution). Above 40 mg/L accuracy decreases and BaSO4 suspensions lose
stability.

Procedure
1. Filter the sample though filter paper (Whatman No. 1) and take 50 mL of filtrate in an
Erlenmeyer flask.
2. Add 20 mL buffer solution and mix in stirring apparatus. While stirring, add 0.15 g of barium
chloride to the sample and stir the sample with the help of magnetic stirrer for about an hour.
3. Measure the absorbance against a distilled water blank (do not add barium chloride to it) at
420 nm using spectrophotometer. Absorbance for the blank sample is taken to correct for sample
color and turbidity.

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 27

4. Process the standard solution of different strengths in similar way and record the absorbance
for each solution. Plot a standard sulfate calibration curve on a graph paper from these
absorbance values putting strengths (mg/L) on X-axis and absorbance @ 420 nm on Y-axis. Fit a
best-fit linear model to the data. Express equation as:
Absorbance value(y) = m× Sulfate concentration (in mg/L) + c
5. Using the standard sulfate calibration curve (a linear-model; Equation), find out sulfate
concentration in the given unknown sample in mg/L.
Sulfate concentration (mg SO42-/L) = (1000×mg SO42-)/(mL sample)

Results: The Sulfate of the given wastewater sample = ----------mg/L.

Environmental Significance:

Exp. No: Date:

Determination of Fluride
Aim:- To determine the Fluoride given the sample by using Ion Selective Electrode methods.

Introduction
 Fluoride is a natural mineral that is found in many foods and in all drinking water. The
amount of fluoride in water varies from area to area.
Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 28
 Fluoride occurs naturally in public water systems as a result of runoff from weathering of
fluoride-containing rocks and soils and leaching from soil into groundwater.
 Atmospheric deposition of fluoride-containing emissions from coal-fired power plants and
other industrial sources also contributes to amounts found in water, either by direct
deposition or by deposition to soil and subsequent runoff into water.

Material and Chemicals Required:-

1. NaF, dried at 100 °C for 1 hour.
2. Liquid NaF unknown.
3. KCl (7.55 g)
4. Fluoride ISE and Ag/AgCl or SCE reference electrode.
5. Multimeter or pH meter capable of displaying mV potentials

Procedure
Preparation of Standards
1. Dry the NaF solid for 1 hour at 100ᵒC.
2. Accurately weigh out about 0.42 g of NaF, dissolve in deionized water, dilute to 100 mL in a
volumetric flask and mix well. This solution is about 10-1 F in NaF.
3. Transfer 10 mL of the solution prepared in (2) to a 100 mL volumetric flask using a pipet,
dilute to volume with deionized water and mix well. This solution is about 10-2 F in NaF.
4. Weigh out 7.55 g of KCl on a top-loading balance and dissolve in 100 mL of deionized water.
This solution is 1 F in KCl.
5. Prepare standard solutions in four 100 mL volumetric flasks as follows:

Dilute each flask to volume with deionized water and mix well.
Determination of unknown samples
1. Add 1 mL of your prepared unknown, then 10 mL of KCl to a 100 mL volumetric flask.
Dilute to the 100 mL mark with deionized water.
2. Measure the potential in mV of the fluoride ISE vs the reference electrode for each of the four
standards and unknowns.
3. Pour about 30 mL of each standard or unknown solution into a clean, dry 100 mL beaker and
immerse the electrodes in the solution to a depth of not more than 2 cm. Measure the electrode
potential, taking care to note both the sign and the magnitude of the potential.

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 29

4. When you finish, rinse the electrodes with deionized water. Leave the reference electrode in
the appropriate storage solution. The F- ISE should be stored dry and loosely capped.

Results: The amount of F- present in the given sample is: _______

Environmental Significance:

Exp. No: Date:

Bacteriological analysis of water by MPN method
Aim: To find the Most Probable Number(MPN) of bacterial density by [Link] test.

Introduction:

Materials Required:
Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 30
Apparatus
Petri dishes,
Fermentation tubes
Autoclaves
Incubator
Test tubes
Pipettes
Measuring jar
Sampling bottle ( sterile)
MacConkey or Lactose broth
EMB agar, Nurtient agar
Durham’s tube
Test tube stand
Water sample
Reagents
1. Lactose broth
2. Lauryl tryptose
3. Brilliant green lactose bile broth
4. Endo agar
5. Eosin methylene blue agar atc.,
Procedure:
for presumptive test
 Prepare MacConkey purple media of single and double strength in test tubes with
Durham’s tube and autoclave it.
 Take three sets of test tubes containing five tubes in each set; one set with 10 mL of
double strength (DS) other two containing 10 mL of single strength (SS).
 Using sterile pipettes transfer 10 mL of water to each of DS broth tubes. Transfer 1 mL of
water sample to each of 5 tubes of one set of SS broth and transfer 0.1 mL water to five
tubes of remaining last set of SS broth tubes.
for presumptive test
 Incubate the tubes at 37°C for 24 hours
 After incubation, observe the gas production in Durham’s tube and color change of the
media.
 Record the number of positive results from each setand compare with standard chart to
give presumptive coliform count per 100 mL water sample.
For confirmed test
 Take the positive tube from the presumptive test and using Eosine Methylene Blue
(EMB) in duplicate.
 Incubate one plate at 37°C for 24 hours and another at 44.5°C for 24 hours.

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 31

  Look for typical colonies in the media; blue black with green metallic sheen colonies are
of E. coli in EMB agar.
Completed test
 Inoculate the colony in a tube of Lactose broth with Durham’s tube.
 Subculture the colony on Nutrient agar plate. This subculture is considered optional.
 Incubate the broth cultures at 37°C and 44.5°C and Nutrient agar at 37°C.
 Examine for acid and gas production in Lactose broth. The nutrient agar is used for Gram
staining and for IMViC (Indole test. Methyl red test. Voges-Proskauer test. Citrate
utilization test.) test.

Computation of MPN

MPN/100 mL = (No. of positive tubes x 100) / Square root of (mL sample in negative tubes x
mL sample in all tubes)

Observation

Results: MPN/100 mL = -------------------

Environmental Significance:
1.
2. etc.,
Exp. No: Date:
Determination of Biochemical Oxygen Demand (BOD)

Aim: To determine the BOD value of the given water sample.

Introduction:

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 32

 The amount of O2 absorbed by a sample of sewage during a specific period, generally 5 days
at a specific temperature, generally 20°C for the aerobic destruction of the organic matter
by living organisms.
 Total BOD is of more significance to food webs than to water quality.

Apparatus Required:
BOD Incubator, Burette & Burette stand, 300 mL glass stopper BOD bottles, 500 mL conical
flask, Pipettes with elongated tips, Pipette bulb, 250 mL graduated cylinders,Wash bottle

Reagents:
Calcium Chloride, Magnesium Sulphate, Ferric Chloride, Di Potassium Hydrogen Phosphate,
Potassium Di Hydrogen Phosphate, Di sodium hydrogen phosphate, Ammonium Chloride,
Manganous sulphate, Potassium hydroxide, Potassium iodide, Sodium azide, concentrated
sulfuric acid, Starch indicator, Sodium thiosulphate, Distilled or deionized.

Procedure:
1. Completely fill two BOD bottles with dilution water.
2. Into additional BOD bottles, partially filled with dilution water, carefully mix the proper
volume of sample.
3. Add dilution water until the bottles are completely filled.
4. Stopper each bottle taking care to avoid trapping air bubbles inside the bottles as the bottle
stoppers are inserted.
5. Fill the top of each bottle neck around the stopper with dilution water.
6. Determine the initial DO content of each set of duplicate bottles, including the dilution water
blank
7. Place the remaining bottles in the incubator at 20°C and incubate for five days.
8. At the end of exactly five days (+/-3 hours), test the DO content of the incubated bottles.
9. Calculate the BOD for each dilution.
10. The dilution water blanks are used only to check the quality of the dilution water.
11. If the quality of the water is good and free from impurities, the depletion of DO should be
less than 0.2 mg/L.
12. In any event, do not use the depletion obtained as a blank correction.

Tabulation
Burette Solution: Sodium Thiosulphate
Pipette Solution: Sample
Indicator: Starch

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 33

End point: Disappearance of blue color

Volume of Burette Reading Volume of Dissolved

Trial Sample (mL) Titrant Oxygen
No. Day (mL) (mL) (mg/L)
Initial Final (Na2S2O3
solution used)

Blank
1.
2.
Blank
1.
2.

Calculation
BOD, mg/L = (Initial DO - Final DO) x bottle volume)/Sample volume (1)
1. Major interferences of BOD test are substances that inhibit the growth of the
microorganisms viz. chlorine, mineral acids, and heavy metals.
2. Excessive nitrites can interfere with the BOD determination.
3. Growth of algae in the presence of light can increase the DO.
4. Artifacts in BOD testing results from residues building up in the BOD and dilution water
bottles.
5. Variations in BOD test is subject to a number of factors temperature, weather etc. results can
vary widely from day to day, or even hour to hour.
Results: The BOD of the given wastewater sample = ----------mg/L.
Environmental Significance:
 Principle test for biodegradability of any sample and strength of the waste so measures the
amount of pollution.
 Important parameter to assess the pollution of surface and ground waters where
contamination occurred due to disposal of domestic and industrial effluents.
 Allows calculation of the effect of the discharges on the oxygen resources of the receiving
water.
 Measurement of BOD in raw (influent) and treated (effluent) wastewaters is a standard
practice to evaluate treatment facility performance.
 Data from BOD tests used for the development of engineering criteria for the design of
wastewater treatment plants.

Exp. No: Date:

Jar Test – Coagulation/Flocculation

Aim:

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 34

1) To understand the process of coagulation and flocculation using alum to remove turbidity of
water.
2) To determine the optimum coagulant dose for a given water sample.

Introduction
 The jar test simulates the coagulation/flocculation process in a batch mode operation.
 A series of batch tests are run in which pH, coagulant type and dosage concentration, etc.,
are varied to get the optimal point and maximum removal turbidity.
 An economic analysis is performed to select these parameters.
 The primary purpose of the coagulation/flocculation process is the removal of turbidity from
the water and wastewater.
 Turbidity is a cloudy appearance of water caused by small particles suspended therein.
 Water with little or no turbidity will be clear.
 Water with a high turbidity need to be removed from water.
 The process removes many bacteria which are suspended in the water and can be used to
remove color from the water.
Methodology
Conducting jar test with variable dosage of alum concentration.

Apparatus required:
Jar test apparatus, Weighing balance, Stop watch, Turbidity meter, 3.5mL, 10 mL pipette, 1000
mL beakers, pH meter, etc.,

Chemicals Required:
Aluminum sulphate (Alum)-(Al2(SO4)3.18H2O)

Procedure:
1. Take 1 L of given water sample in 6 beakers.
2. Find the pH of the sample and adjust it to 6 to 8.5.
3. Now add various doses of Alum (i.e) 1 g, 2 g, 3 g, 4 g and 5 g, and 6 g to different beakers.
4. Now insert the paddle of the jar testing apparatus inside the beaker and start it.
5. Initially maintain a speed such that the paddle rotate at an angular speed 300 rpm for a time 10
minutes
6. Now adjust the speed such that the paddles rotate at 100 rpm for a time of 15 minutes.
7. Now allow the beakers to settle down for 10 minutes.
8. Make an observation as of which of the 6 beakers is most clear. Also measure the turbidity of
each beakers using turbidity meter and tabulates you results.
9. Plot graph settled turbidity Vs coagulant dosage.

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 35

Tabulation

Dosage , Turbidity, Turbidity, Final, Removal Settlement,

g/L Initial, mg/L mg/L efficiency , (mm)
%
1
2
3
4
5

Removal efficiency = ((Turbidityinitial - Turbidityfinal )/ Turbidityinitial)*100 (1)

Results:
Optimum dosage of coagulant is --------- mg/L

Environmental Significance:
 Excess dosage of alum may contribute excess aluminum in drinking water.
 Less dosage of Alum do not remove turbidity in water, which ultimately increase load on
filters. So the optimum dosage should be added in coagulation process to prevent the above
filtration.
 Coagulation removes not only turbidity but also color, microorganism, algae, phosphate, taste
and odor, etc.,

Dr. P. Asaithambi/Assistant Professor/WSEE/JiT/JU Page 36