Bharati Vidyapeeth's College of

Engineering
A-4, Paschim Vihar, New Rohtak Road, New Delhi-110063

LAB MANUAL

Department of Applied Science

SUBJECT NAME: ENVIRONMENTAL STUDIES LAB

SUBJECT CODE: BS-161/162

 Vision & Mission of Institute
Vision of t h e Institute: To be an institute of excellence that provides quality technical
education and research to create competent graduates for serving industry and society.

Mission of the Institute:

M1: To impart quality technical education through dynamic teaching-learning environment.

M2: To promote research and innovations activities which gives opportunities for life-long
learning in context of academic and industry.

M3: To build up links with industry-institute through partnerships and collaborative

developmental works.

M4: To inculcate work ethics and commitment in graduates for their future endeavours to
serve the society

Vision & Mission of Department

Vision of the Department:

The department aspires to be a center of excellence in education in basic sciences and technology
with ethical and social values.

Mission of the Department:

DM-1: To provide quality education through professional, problem-driven and interdisciplinary

teaching methodology

DM-2: To make students sensible in terms of ethical and social values in pursuing their
education
 Program Outcomes (POs)

1. Engineering knowledge: Apply the knowledge acquired in mathematics, science, engineering for
the solution of complex engineering problems.

2. Problem analysis: Identify research gaps, formulate and analyze complex engineering problems
drawing substantiated conclusions using basic knowledge of mathematics, natural sciences and
engineering sciences.

3. Design/development of solutions: Design solutions for the identified complex engineering

problems as well as develop solutions that meet the specified needs for the public health and safety,
and the cultural, societal, and environmental considerations.

4. Conduct investigations of complex problems: Use research-based knowledge and research

methods, including design of experiments, analysis and interpretation of data and synthesis of the
information to provide valid conclusions.

5. Modern tool usage: Work on the latest technologies, resources and software tools including
prediction and modelling to complex engineering activities with an understanding of their limitations.

6. The engineer and society: Apply the basic acquired knowledge to measure societal, health, safety,
legal and cultural issues and identifying the consequential responsibilities relevant to the professional
engineering practice.

7. Environment and Sustainability: Comprehend the impact of the professional engineering

solutions in context of society and environment and demonstrate the need and knowledge for
sustainable development

8. Ethics: Apply ethical principles and commit to professional ethics and responsibilities and norms
of the engineering practice

9. Individual and team work: Function effectively as an individual, and as a member or leader in
diverse teams, and in multidisciplinary settings.

10. Communication: Communicate effectively on complex engineering activities with the

engineering community and with society at large, such as, being able to comprehend and write effective
reports and design documentation, make effective presentations, and give and receive clear instructions.

11. Project management and finance: Demonstrate knowledge and understanding of the engineering
and management principles and apply these to one’s own work, as a member and leader in a team, to
manage projects and in multidisciplinary environments.

12. Life-long learning: Recognize the need for, and have the preparation and ability to engage in
independent and life-long learning in the broadest context of technological change.
 Subject Syllabus/Lab Plan

Course Code(s): BS 161/162 Credits: 1

Practical: 2 periods/week internal assessment: 40 marks
Paper: Environmental Studies Lab External assessment: 60 marks
PaperCode: BS-161 /BS- Paper: Environmental Studies Lab. L P C
162
- 2 1
Marking Scheme:
1. Teachers Continuous Evaluation: 40 marks
2. Term end Theory Examinations: 60 marks
Instructions:
1. The course objectives and course outcomes are identical to that of “Environmental
Studies” as this is the practical component of the corresponding theory paper.
2. The practical list shall be notified by the teacher in the first week of the class
commencement under intimation to the office of the office of the Head of Department
/ Institution in which the paper is being offered from the list of practical below. At
least 8 experiments must be performed by the students

LIST OF EXPERIMENTS

1. Determination of pH, conductivity and turbidity in drinking water sample.

2. Determination of pH and conductivity of soil/sludge samples.
3. Determination of moisture content of soil sample.
4. Determination of Total Dissolved Solids (TDS) of water sample.
5. Determination of Dissolved Oxygen (DO) in the water sample.
6. Determination of Biological oxygen demand (BOD) in the water sample.
7. Determination of Chemical oxygen demand (COD) in the water sample.
8. Determination of Residual Chlorine in the water sample.
9. Determination of ammonia in the water sample.
10. Determination of carbon dioxide in the water sample.
11. Determination of nitrate ions or sulphate ions in water using spectrophotometer.
12. Determination of the molecular weight of polystyrene sample using viscometer method.
13. Base catalyzed aldol condensation by Green Methodology.
14. Acetylation of primary amines using eco-friendly method.

15. To determine the concentration of particulate matter in the ambient air using High Volume
Sampler.
References:

1. Vogel’s Text Book of Quantitative Chemical Analysis by G.H. Jefferey, J. Bassett, J. Mendham, and
R.C. Denney, Logmaan Scientific & Technical, 1989
2. Essentials of Experimental Engineering Chemistry by S. Chawla, Dhanpat Rai & Co., 2008.
3. Experiments in Applied Chemistry by S. Ratan, S.K. KAtaria & Sons, 2003.
4. Practical Chemistry by O.P.Pandey, D. N. Bajpai and S. Giri, S.Chand & Co., 2005.
5. Engineering Chemistry with Laboratory Experiments by M. S. Kaurav, PHI Learning Pvt. Ltd., 2011.

6. Laboratory Manual on Engineering Chemistry by S. K. Bhasin, and Sudha Rani, Dhanpat Rai
&Co., 2006.

Note:

1. For better understanding of various aspects of environment visits to local areas, depending upon
easy access and importance may be planned to any nearby river, forest, grassland, hills and students
should write a report based on their observations.
2. At least 8 Experiments out of the list shall be performed by the students. Teachers may introduce
new experiments for the class in addition to above.
TITRATION:

The word titration comes from the Latin word "titulus", which means inscription or title. The
French word title means rank. Therefore, Titration means the determination of concentration or
rank of a solution with respect to water with a pH of 7. The standard solution is usually added
from a graduated vessel called a burette. The process of adding standard solution until the
reaction is just complete is termed as titration and the substance to be determined is said to be
titrated.
All chemical reactions cannot be considered as titrations. A reaction can serve as a basis of a
titration procedure only if the following conditions are satisfied:
1. The reaction must be a fast one.
2. It must proceed stoichiometrically.
3. The change in free energy (ΔG) during the reaction must be sufficiently large for
spontaneity of the reaction.
4. There should be a way to detect the completion of the reaction.

End point and Equivalent point:

For a reaction, a stage which shows the completion of a particular reaction is known as end
point. Equivalence point is a stage in which the amount of reagent added is exactly and
stoichiometrically equivalent to the amount of the reacting substance in the titrated solution.
The end point is detected by some physical change produced by the solution, by itself or more
usually by the addition of an auxiliary reagent known as an 'indicator'. The end point and the
equivalence point may not be identical. End point is usually detected only after adding a slight
excess of the titrant. In many cases, the difference between these two will fall within the
experimental error.

Indicator:

It is a chemical reagent used to recognize the attainment of end point in a titration. After the
reaction between the substance and the standard solution is complete, the indicator should give
a clear color change. When a titration is carried out, the free energy change for the reaction is
always negative. That is, during the initial stages of the reaction between A & B, when the titrant
A is added to B the following reaction takes place.
 Equilibrium constant,

a = activity co-efficient.

Large values of the equilibrium constant K implies that the equilibrium concentration of A & B
is very small at the equivalence point. It also indicates that the reverse reaction is negligible and
the product C & D are very much more stable than the reactants A & B. Greater the value of K
the larger the magnitude of the negative free energy change for the reaction between A & B.
Since,

Where,
R= Universal gas Constant = 8.314 JK-1mol-1, T= Absolute Temperature

The reaction of the concentration of A & B leads to the reduction of the total free energy change.
If the concentrations of A & B are too low the magnitude of the total free energy change
becomes so small and the use of the reaction for titration will not be feasible.

Expressions of Concentration of Solutions:

The concentration or strength of solution means the amount of solute present in a given amount
of the solution. The concentration may be expressed in physical or chemical units.
1. Normality (N): It is defined as number of gram equivalents of the solute present in 1 litre
(1000mL.) of the solution. If W g of solute of equivalent weight E is present in V mL of
the solution, the normality of the solution is given by:

2. Molarity (M): It is defined as the number of moles of the solute present in 1 liter (or 1000
mL) of the solution. A one molar solution contains 1 mole of the solute dissolved in 1 liter
of the solution.
3. Molality (m): It is defined as the number of moles of solute dissolved in 1000 g of the
solvent. One molal solution contains one mole of the solute dissolved in 1000 g of the
solvent.
 4. Normal solution: A solution containing one-gram equivalent weight of the solute
dissolved per litre is called a normal solution; e.g. when 40 g of NaOH are present in one
litre of NaOH solution, the solution is known as normal (N) solution of NaOH. Similarly,
a solution containing a fraction of gram equivalent weight of the solute dissolved per litre
is known as subnormal solution. For example, a solution of NaOH containing 20 g (1/2
of g eq. wt.) of NaOH dissolved per litre is a sub- normal solution. It is written as N/2 or
0.5 N solution.
Formulae used in solving numerical problems on volumetric analysis;
1. Strength of solution = Amount of substance in g litre-1.
2. Strength of solution = Amount of substance in g moles litre-1.
3. Strength of solution = Normality × Eq. wt. of the solute = molarity × Mol. wt. of solute.
4. Molarity = Moles of solute/Volume in litre.
5. Number of moles = Wt.in g/Mol. wt = M × V (initial) = Volume in litres/22.4 at NTP
(only for gases).
6. Number of milli moles = Wt. in g × 1000/mol. wt. = Molarity × Volume in mL.
7. Number of equivalents= Wt. in g/Eq. wt. = x × No. of moles × Normality × Volume in
litre(Where x = Mol. wt./Eq. wt.).
8. Number of mill equivalents (meq.) = Wt. in g × 1000 / Eq. wt. = normality × volume in
mL.
9. Normality = x × No. of mill moles (Where x = valency or change in oxidation number).
10. Normality formula, N1V1 = N2V2, (Where N1, N2 → Normality of titrant and titrate
respectively, V1, V2 → Volume of titrant and titrate respectively).
11. % by weight = Wt. of solvent/Wt. of solution ×100.

A solution is a homogeneous mixture of two or more components, the composition of which may
be changed. The substance which is present in smaller proportion is called the solute, while the
substance present in large proportion is called the solvent.

Volumetric Analysis:
It involves the estimation of a substance in solution by neutralization, precipitation, oxidation or
reduction by means of another solution of accurately known strength. This solution is known as
standard solution.
Volumetric analysis depends on measurements of the volumes of solutions of the
interacting substances. A measured volume of the solution of a substance A is allowed to react
completely with the solution of definite strength of another substance B. The volume of B is
noted. Thus we know the volume of the solutions A and B used in the reaction and the strength
of solution B; so the strength of the other solution A is obtained. The amount (or concentration)
of the dissolved substance in volumetric analysis is usually expressed in terms of normality. The
weight in grams of the substance per liter of the solution is related to normality of the solution as,
Weight of the substance (g per liter) = Normality × gram equivalent weight of the substance.

Conditions of Volumetric Analysis:

1. The reaction between the titrant and titrate must be expressed.

2. The reaction should be practically instantaneous.

3. There must be a marked change in some physical or chemical property of the solution at
the end point.

4. An indicator should be available which should sharply define the end point.
 Practical Manual
EXPERIMENT-1

AIM: To estimate the soil moisture content in the given soil sample.

APPARATUS: Watch glass; Drying oven, Spatula, Desiccator

THEORY: Water plays a very significant role in soil growth relationship. It forms major part
of plant itself, essential for physiological activities, acts as a solvent and nutrient carrier and
maintains turgidity of the plants. In fact water is a regulator of physical, chemical and biological
activities in the soil.
Water in a soil may be measured in a number of ways: Gravimetric, Volumetric method,
Neutron scattering, Soil moisture tension and Electrical conductivity. However gravimetric
method is the simplest and the most widely used method. In this the weighed Soil sample is placed
in an oven at 105oC and it is dried up to constant weight. The weight difference is considered to
be water present in soil sample.

Percent moisture = (Loss in weight /weight of soil) × 100

PROCEDURE
1. Weigh the empty watch glass.
2. Put 10 g soil sample on the watch glass and weigh immediately to avoid loss in moisture
content.
3. Place the watch glass in a preheated oven at 105oC for 1 h.
4. Keep the watch glass in a desiccator for cooling for at least half an hour.
5. Now weigh the watch glass along with the oven dry soil.
OBSERVATIONS

Weight of empty watch glass = x grams

Weight of watch glass + moist soil = y grams

Weight of watch glass + oven dried soil = z grams

CALCULATIONS

Moisture content in soil = (y - z)

Weight of moist soil = (y - x)

Percentage moisture in soil = (y - z) / (y - x) × 100.

RESULT: The amount of moisture present in soil sample is………..

 EXPERIMENT-2

AIM: Determination of Total dissolved solids in water/ effluent sample.

THEORY: Dissolved solids" refer to any minerals, salts, metals, cations or anions dissolved in
water. Total dissolved solids (TDS) content of drinking water varies between 20-1000 mg L-1, and
it comprises mainly of inorganic salts (principally calcium, magnesium, potassium, sodium,
bicarbonates, chlorides and Sulphates) and some small amounts of organic matter and dissolved
gases.
TDS in drinking-water originates from natural sources, sewage, urban runoff, industrial-
wastewater, and chemicals used in the water treatment process, and the nature of the piping or
hardware used to convey the water, i.e., the plumbing.

Table 1: Total Dissolved Solids (TDS) in some water samples.

Water Type TDS (mg L-1)

Sea Water 35,000
Potable Water <500
Irrigation Water 0-2000

PROCEDURE

1. Take the empty weight of evaporating dish/crucible.

2. Measure accurately 100ml of well mixed water/effluent Sample and filter the sample
through the whatman filter paper.
3. Transfer filtrate to a preweighed evaporating dish/crucible and evaporate to dryness on a
hot plate.
4. Dry for at least 1 h in oven at 180oC.
5. Cool in a desiccator and take the final weight. Calculate the total dissolved solids (TDS)
using the given equation.

CALCULATION:

Total dissolved solids (mg /L) = (𝐴 − 𝐵) ×

.

Where A = weight of dried residue + dish

B= weight of dish in mg
V = volume of sample in litre

RESULT: The amount of total disso1ved solids in a given water sample is …….mg/L
SIGNIFICANCE: An elevated TDS indicates the following:
1. The concentration of the dissolved ions may cause the water to be corrosive, salty or
brackish taste, results in scale formation, and interfere and decrease efficiency of hot water
 heaters; and
2. Many contain elevated levels of ions that are above the Primary or Secondary Drinking
Water Standards, such as: an-elevated level of nitrate, arsenic, aluminium, copper, lead,
etc.
DISSOLVED OXYGEN AND OXYGEN DEMAND
One of the most important water quality parameter is the amount of dissolved oxygen (DO) present. Oxygen is
poorly soluble in water. The amount of oxygen in water depends on physical, chemical and biochemical
activities taking place in water. The solubility of dissolved oxygen in water at saturation at any temperature and
pressure is given by Henry’s law. The – saturation value of DO in water is of the order of 8-15 mgL-1, depending
upon temperature. The minimum amounts required for aquatic life for a healthy fish population is 6-8 mgL-1.

Significance of the determination of DO

 In aquatic life, as DO drops, fish and other species are threatened and may get killed.
 Fall in DO levels causes undesirable odours, tastes and reduce the acceptability of water
for domestic use.
 In steam generation, DO is one of the most important factors causing corrosion of the
boiler material.

There are several measures of oxygen demand commonly used. Two of these are:

(i) Biochemical oxygen demand (BOD)

(ii) Chemical oxygen demand (COD)

BIOCHEMICAL OXYGEN DEMAND

When biodegradable organic matter is released into water, the microorganisms feed on the wastes and break it
down into simpler organic or inorganic substances. If the decomposition takes place in the presence of oxygen
i.e aerobically the non-objectionable, stable end products are formed.

On the other hand if insufficient oxygen is available, the decomposition takes place anaerobically. The micro-
organisms causing the decomposition of the organic matter in absence of oxygen are entirely different from
aerobic bacteria and produce highly objectionable end products including H2S, NH3 and CH4.

𝑂𝑟𝑔𝑎𝑛𝑖𝑐 𝑀𝑎𝑡𝑡𝑒𝑟 + 𝑂2 → 𝐶𝑂2 + 𝐶𝐻4 + 𝑂𝑡ℎ𝑒𝑟 𝑢𝑛𝑠𝑡𝑎𝑏𝑙𝑒 𝑝𝑟𝑜𝑑𝑢𝑐𝑡𝑠

The amount of oxygen required by micro-organisms to oxidize organic wastes aerobically is called
biochemical oxygen demand (BOD).
Actually when sufficient O2 is present, i.e. aerobic systems, oxygen is the oxidizing agent to convert organic
waste into CO2 and H2O. Similarly the inorganic materials also oxidized. The products obtained are not harmful.
However, if sufficient oxygen is not present (anaerobic conditions), organic matter is oxidized by nitrate to form
undesirable products.
5CH 3COO - + 8NO 3-  8H + 
Bacteria
 5CO 2  5HCO3  4N 2  9H 2O
CH 3COO - + NO3- 
Bacteria
 CO 2  HCO3  NH 3

The products in this case like NH3 etc. give bad odour to water.
Example NH3 give smell of rotten fish. Similarly under anaerobic conditions, SO 24  , PO34 , CO2 are reduced to

H2S, SH-, phosphine, methane etc. giving rotten egg smell or suffocating smell.
Therefore oxygen demanding wastes are termed as water pollutants because they deplete its dissolved oxygen.
Thus sufficient oxygen may be present or supplied to maintain aerobic conditions and hence prevent bad odour;
taste and colour.

CHEMICAL OXYGEN DEMAND

Although BOD test is applicable to organic wastes, there are many drawbacks. Certain organic materials are
not biodegradable and hence can give wrong conclusions that less organic matter is present because BOD of
such water samples will be low. In this case COD, Chemical Oxygen Demand reveals the real organic content
present. Here the oxidation of organic substances present in water is done chemically.
COD, is the amount of oxygen required by organic matter in a sample of water for its oxidation by a strong
chemical oxidizing agent such as K2Cr2O7.
Since in COD determination, the organic matter, both biologically oxidizable (glucose) and biologically inert
(cellulose) is completely oxidized to CO2 and H2O; COD values are greater than BOD values.

EXPERIMENT NO. 3
AIM: Determination of dissolved oxygen in sample of water.
APPARATUS: Burette, Pipette, Conical flask, Beaker etc.
CHEMICALS: Na2S2O3 (N/40), MnSO4 solution, KI, Starch, Conc.H2SO4.
THEORY: Oxygen itself is not a pollutant in water but its deficiency is an indicator of several types of pollution
in water. Dissolved oxygen (DO) is determined by Winkler’s method or iodometric titration. The dissolved
oxygen in water oxidizes KI and an equivalent amount of iodine is liberated. This iodine is titrated against a
standard hypo solution. However since dissolved oxygen in water is in molecular state and is not capable of
reacting with KI, therefore an oxygen carrier such as manganese hydroxide is used.
The method involves introducing a conc. solution of MnSO4, NaOH and potassium iodide, azide reagent, into
the water sample. The white precipitate of Mn(OH)2 which is formed, is oxidized by oxygen in water sample

to give a brown precipitate of basic manganese oxide.

This MnO(OH)2 , in acidic medium dissolves and liberates free iodine from the added KI in an equivalent
amount of dissolved oxygen in water sample. This liberated I2 is then titrated against Na2S2O3 solution using
starch as indicator. The reactions involved are:
MnSO4  2NaOH  Mn(OH)2 +Na 2SO4
4Mn(OH)2 +O2  2MnO (OH)2
2MnO (OH)2  H2SO4  MnSO4  2H 2O +O
2KI  H 2SO4  O  K 2SO4  H2 O +I2
2Na 2S2O3  I2  Na 2S4O6  2NaI

The nitrites present in water, interfere with the titration as these can also liberate I2 from KI.
2HNO2  H 2SO4  2KI  2NO  K 2SO4  2H 2O  I2
Thus to destroy nitrite, sodium azide is used.
2NaN3  H 2SO 4  2HN3  Na 2SO 4
HNO 2  HN3  NO 2  N 2  H 2 O

PROCEDURE:
1. A known amount of sample water (say 250 ml) is taken in a stoppered bottle avoiding contact with air: -
2. Add 0.4 ml of MnSO4 solution to it by means of a pipette, dipping the end well below the surface of water.
Also add 2 ml of alkaline iodide-azide solution to it.
3. Stopper the bottle and shake thoroughly. Allow the brown precipitates of MnO(OH)2 formed to settle down.

4. When some portion of the liquid below the stopper is clear, add 2 ml of conc. H2SO4 with the help of pipette.
Stopper and mix till the precipitate is completely dissolved. The characteristic brown colour of iodine is
produced.
5. Transfer 100 ml of the above solution in a 250 ml flask with a pipette. Titrate the liberated I2 with standardized
sodium thiosulphate solution until the sample solution becomes pale yellow.
6. Add 2 ml of starch solution, the solution will turn blue.
7. Continue titration till the blue colour disappears.
8. Repeat to get concordant reading.
OBSERVATIONS
Volume of the water sample taken for titration = 100 ml
Observation Table:

S. No. Burette Readings Volume of the titrant

Volume of the solution taken in the Initial Final used
titration flask Reading Reading (Final-Initial reading)

1
2
3

CALCULATIONS
Hypo = Oxygen in water

N1V1 = N2V2

N1  V1  N 2  100
N1V1
N2 
100
Strength of dissolved oxygen = N2  Equivalent Weight

N1V1
  8gL1
100
N1V1
  8 1000 mgL1
100
RESULT: The amount of dissolved oxygen in water =……ppm
PRECAUTIONS:
1. The water should be taken in the stoppered bottle very carefully without trapping air bubbles which could
raise oxygen level by aerating the sample.
2. MnSO4 and alkaline iodine-azide solutions are added to the water sample just below the surface of water.
3. Whole of the precipitate of MnO(OH)2 should be dissolved in H2SO4.

Instructions for the Lab Assistant:

1. MnSO4 solution: Dissolve 50g of MnSO4 in distilled water and make up the volume to 1 L.
2. Alkaline potassium iodide azide reagent: Dissolve 40 g of NaOH, 20 g of KI in distilled water-, add 0.5
g of reagent grade sodium azide to the cooled solution and make up the volume to 100 ml
3. N/40 Na2S2O3 solution: Dissolve 6.20 g of Na2S2O3 in distilled water and make up this volume to 1 L.
Standardize it with N/40 K2Cr2O7 solution.
Viva Questions:

1. Why MnSO4 is used in the above determination?

Ans. Since oxygen in water is in molecular form and is not capable of reacting with KI, therefore an oxygen
carrier such as manganese hydroxide is used. This in turn is prepared by MnSO4 and NaOH.
2. Why azide is added to the reaction mixture?
3. What is the significance of the above determination?
Ans. Measurement of D.O. is necessary because
 It causes corrosion in boiler feed waters.
 Conc. of dissolved oxygen is a good indicator for polluted Waters.
 DO is necessary for life of fish and other aquatic organisms.
Note: Sodium azide is poisonous. It may also explode if exposed to heat.
 EXPERIMENT No. 4

AIM: To determine Biological Oxygen Demand (BOD) of a given sample of water.

APPARATUS: Burette, Pipette, Conical flasks, Beakers.
CHEMICALS: MgSO4 solution (22.5 g/L), CaCl2 (27.5 g/L), FeCl3 solution (0.25 g/L), phosphate buffer
Solution; BOD bottles, Na2S2O3.
THEORY: The BOD in a water sample is directly related to the extent of pollution by sewage or other oxygen
demanding waste. If the oxygen in water is less than Biological Oxygen Demand, the decomposition of
microorganisms-takes-place-anaerobically (in absence of oxygen) and hence produce highly objectionable end
products including H2S, NH3 and CH4. Therefore sufficient oxygen must be present or supplied externally to
maintain aerobic conditions so that organic matter decomposes in to acceptable products. To know the amount
to oxygen required by the given Sample, the determination of BOD is necessary.
Polluted waters will continue to absorb oxygen for many months, till the oxidation gets completed and hence it
is not practical to determine this ultimate oxygen demand which would require an extended period of time. As
a result, it has become standard practice to simply measure the oxygen over a shorter period of 5 days. This is
known as 5-day BOD test. BOD5 is the total amount of oxygen consumed by micro-organisms during the first
five days of biodegradation.
In its simplest form BOD5 would involve diluting a known volume of sample water with a known
volume of pure water, whose oxygen content is already known: Let it be D1. This is kept for 5 days at 20 oC in
a stoppered bottle, away from light. The dissolved oxygen is measured after the period of incubation. Let it be
D2. The difference between the original oxygen content and residual oxygen content will indicate the oxygen
consumed by the water sample in five days in oxidizing the organic matter present in water sample. Thus BOD5
is given by:
Volume of sample after dilution
BOD5  D1 -D 2
Volume of sample before dilution

Where, D1 = D.O. of the sample in mg L-1 at the start of experiment

D2 = D.O. of the sample in mg L-1 after 5 days.
= Loss of oxygen in mg L-1 × Dilution factor
The dilution of the waste water sample with pure water is necessary because waste water contains more of
oxygen demanding material and hence the dissolved oxygen may not be sufficient to decompose it aerobically.

PROCEDURE:
Preparation of dilution water:
1. Take 1000 ml of distilled water in bottle. Add l ml each of phosphate buffer (pH = 7.2), CaCl2 (27.5 g L-1)
MgSO4 (22.5 g L-1) and FeCl3 (0.25 g L-1).
2. Water is aerated by passing compressed air through diffusion tube until it is completely saturated.
Determination
3. Take about 5 ml of the .sample and dilute it to 300 ml with dilution water prepared above (or any appropriate
dilution).
4. The diluted sample is taken in two bottles filled up to the neck.
5. The dissolved oxygen in one bottle is determined immediately and in the other bottle after five days of
incubation. The procedure for determining the DO is the same given in experiment number 3.
OBSERVATION

(i) Determination of. D. O. in bottle No. 1

S. No. Volume of the solution taken in Burette Readings Volume of the

titration flask (ml) Initial Final titrant used (final-
Reading Reading initial reading) (ml)
1
2
3

(ii) Determination of. D. O. in bottle No. 2 (after incubation for 5 days)

S. No. Volume of the solution taken in Burette Readings Volume of the

titration flask (ml) Initial Final titrant used (final-
Reading Reading initial reading) (ml)
1
2
3

Volume of sample taken for each titration = 100 ml

N
Normality of Na2S2O3 used =
100
CALCULATIONS:
Volume of the sample before dilution = 5 ml.
Volume of the sample after dilution = 300 ml.

(i) D.O in bottle -1

Hypo = O2 in sample
N1V1 = N2V2
 1
 V1  N 2 100
100
V1
N2 
10000
Strength of dissolved O2 = N2 × Equivalent Weight
V1
N2   0.8g L1  0.8 V1 ppm
10000
(ii) D.O in bottle-2

Similarly find D.O in bottle-No-2

Let it be D2
Volume of sample after dilution
(iii) BOD  D1 -D 2  mgL1
Volume of sample before dilution
RESULT: BOD of the given water sample =…... ppm
Instructions for the Lab Assistant
Prepare the solutions as in the case of determination of dissolved oxygen in experiment no 3.
Viva Questions:
1. What do you mean by BOD?
2. What is BOD5 test?
3. What is the significance of BOD determination?
4. Why bad odour is developed in water if insufficient dissolved oxygen is present in water?
5. How is diluted water prepared?
Ans. It is prepared from distilled water by seeding with a small amount of domestic waste water to provide
a mixed population of bacteria or it can be prepared as given in the experiment.
6. Why, while preparing diluted water is added, 1 ml each of phosphate, FeC13, CaC12, MgSO4 etc.?
Ans. Phosphate is added to maintain a pH of 7 end it also serves as a nutrient. Salt like FeC13 is added to
supply nutrients to bacteria and Ca2+ Mg2+ is added to serve as trace elements needed for growth of bacteria.
Note. The dilution depends upon the nature of the sample. For example 1-4% for industrial wastes, 1-5% for raw and
settled sewage, 25-100% for polluted water, 2-5% for domestic waste water).
 EXPERIMENT NO. 5
AIM: To find the chemical oxygen demand (COD) of a waste water sample using K2Cr2O7
APPARATUS: Burette, pipettes, conical flask, beakers, round bottom flask, condenser, water bath
CHEMICALS: N/4 K2Cr2O7, N/4 Mohr's salt solution, ferroin indicators, sample solution, HgSO4, silver
sulphate reagent.
THEORY: The chemical oxygen demand is defined as the amount of oxygen used while oxidizing the organic
matter content of a sample with a strong chemical oxidant-under-acidic-conditions.
A known amount of K2Cr2O7 is added to the measured amount of the sample and mixture is boiled with conc.
H2SO4. The organic matter gets completely oxidized to produce CO2 and H2O.

 y z y
C x H y O z   x    O 2  xCO 2  H 2 O
 4 2 2

The excess of K2Cr2O7 remained after the reaction is titrated against standard Mohr's salt-solution using ferroin
as indicator. The dichromate consumed is calculated which gives the oxygen required for the oxidation of
organic matter.

INDICATOR: Ferroin

END POINT: Blue to wine red or (colourless to red)

PROCEDURE:

1. Take 50 ml of the sample in a round bottomed flask and add 1 g of HgSO4 and some broken porcelain pieces.
2. Immerse the flask in cold water and slowly add 75 ml silver sulphate reagents with continuous shaking. (As
this is an exothermic process that is why the flask is immersed in cold water)
3. Now add 25 ml of K2Cr2O7 (0.25 N) to this solution and mix the contents of the flask.
4. Attach the reflux condenser and reflux for 2 h.
5. Wash the condenser with distilled water in to the flask. Cool and dilute to about 300 ml by distilled water.

Notes 1. HgSO4 is used to remove the chloride ions which may interfere because of their oxidation under the
experimental conditions.
2. Ag2SO4 is used as a catalyst to achieve the oxidation of straight-chain aliphatic hydrocarbons and low molecular
weight fatty acids.

6. Add 2-3 drops of ferroin as indicator and titrate against 0.25 N Mohr’s salt solution till the end point, i.e.
from blue to wine red.
7. Record the volume of Mohr’s salt solution used. Let it be X ml.
8. Perform a blank titration using distilled water in place of sample solution. For this, take 50 ml of distilled
water in round bottom flask, add the same amount of reagents and reflux for 2 h. Titrate it in the same way as
is done with the sample water.
9. Record the volume of Mohr's salt solution used. Let it be Y ml.
OBSERVATIONS:
Volume of the sample taken = 50 ml.
Observation Table
(a) For sample solution

S. N. Volume of the solution Burette Readings Volume of the

taken in the titration flask titrant used (final-
(ml) Initial Final Reading initial reading) (ml)
Reading
1
2
3

(b) For distilled water (blank titration)

S. N. Volume of the solution Burette Readings Volume of the

taken in the titration flask titrant used (final-
(ml) Initial Final Reading initial reading) (ml)
Reading
1
2
3

Volume of Mohr's salt so1ution used in case of sample solution = X ml (say)

Volume of Mohr's salt solution used in case of distilled water = Y ml (say)
Volume of K 2Cr2O7 used to oxidize 
 the organic matter present in the water sample    Volume of Mohr's salt solutionequivalent to K 2 Cr2 O7   [Y-X]ml
 

N1V1 =N2 V2
(Sample) (Mohr’s salt solution)

1
N1  50 = (Y-X)
4
1
N1 = (Y-X)
4  50
Therefore, COD = N1 × Equivalent Weight of O = N1 × 8 g L-1
RESULT: The. COD of' the given sample is N1 × 8 × 1000 mg L-1
PRECAUTIONS:
1. The addition of Ag2SO4-H2SO4 to the sample should be done slowly with shaking and cooling during mixing.
2. It should always be added through the open end of the condenser.
3. Smaller volume of the sample should be taken.
Instructions to the Lab Assistant
Provide the following:
N
1. K2Cr2O7 solution: Dissolve 12.25g of K2Cr2O7 in distilled water and make up the volume to 1000
4
ml.
N
2. Ferrous Ammonium sulphate (Mohr's salt solution):- Weigh 98 g of Mohr's salt and transfer in flask
4
having a few drops of conc. H2SO4. Add distilled water to dissolve it. Make up the volume to 1000 ml.
3. Ferroin indicator: It can be purchased from the market as such or can be prepared in the lab. Dissolve
3.7125 g of o-phenanthroline monohydrate with 1.6375 g of FeSO4. 7 H2O in distilled water and make
up the volume to 250 ml.
4. HgSO4 (solid)
5. Silver sulphate reagent: Dissolve 5g of AgSO4 in 500 ml of conc. H2SO4.
Viva Questions:

1. Define COD?
2. What is the significance of COD determination?
Ans. The COD is widely used for measuring the polluting properties of domestic and industrial wastes.
3. What is the structure of ferroin indicator?
Ans. Ferroin is a ferrous complex of the organic compound 1, 10-phenanthroline.

1, 10- Phenanthroline Ferrous 1, 10-Phenanthroline (Ferroin)

 EXPERIMENT NO. 6

RESIDUAL CHLORINE

Ingestion of pathogens (disease causing bacteria) not only in drinking water, but also from water that makes it
into person’s mouth through food, utensils and hands.
The process of destroying these pathogenic bacteria and micro-organism is known as sterilization or
disinfection. The chemicals used for the above purpose are called disinfectants. The various methods used for
disinfection of water include, boiling, addition of bleaching powder, chlorine, UV radiation etc. However
chlorination of water is widely used for disinfecting water supply since chlorine is a powerful oxidizing agent
and is cheaply available.
Chlorine is applied to water in its elemental form or as hypochlorite (bleaching powder).

Disinfecting Action of Chlorine: Cl2 reacts with H2O to form hypochlorous acid or hypochlorite ions which
have immediate disastrous effect on micro-organisms.
Cl 2 +H 2 O 
pH < 5
 HOCl+HCl

The hypochlorous acid is unstable and may dissociate as

HOCl 
pH > 8
pH < 7
 H + + OCl-
Hypochlorite ion

All the three forms viz. HOCl, OCl- and Cl2 existing in sample water are termed as free chlorine.
When HOCl react with NH3 likely to be present in water, it forms chloramines.
NH 3  HOCl  NH 2 Cl+H 2 O
NH 2 Cl + HOCl  NHCl 2 +H 2 O
NHCl 2  HOCl  NCl3 +H 2 O
These chloramines also have disinfecting properties and act as chlorine in this form is called combined chlorine.
The chlorine is added to water up to the break point. The break point chlorination of water is defined as
chlorination to such an extent that micro-organisms as well as bad tastes and odour present in water are
destroyed.
However, excess of free chlorine in drinking water is undesirable. An overdose of chlorine imparts an
unpleasant tastes and is also injurious to health.
Hence chlorine determination is necessary in those samples of water that have been treated with chlorine
or hypochlorites (bleaching powder) so that necessary adjustments in dose rate can be made accordingly.
 EXPERIMENT
AIM: Determination of total residual chlorine in water.
APPARATUS: Burette, Pipette, Conical flask and Beakers.
CHEMICALS: N/50 Na2S2O3, Glacial acetic acid, KI, conc. HCl
THEORY: The estimation of residual Cl2 is based on oxidation of KI by residual chlorine i.e. by iodometric
titration.
When the wafer sample containing residual chlorine is treated with KI, residual chlorine (both free and
combined) oxidizes KI and liberates I2 in equivalent amount.
Cl2 +2KI  2KCl+I 2
OCl- +2KI  2KCl+I 2 +H 2 O
HOCl+2KI+HCl  2KCl+I2 +NH 3
NHCl 2 +2KI+2HCl  2KCl+I2 +NH3
The liberated I2 is titrated against hypo solution, using starch as indicator.
I 2  2Na 2S2 O3  2NaI+Na 2S4 O 6
I 2  Starch  Deep blue complex

The end point is disappearance of blue colour.

PROCEDURE:
1. Pipette out 100 ml of the given water sample in conical flask and add 2g of KI solution and about 3 ml
of glacial acetic acid to maintain pH up to 3-4.
2. Cover the flask and shake it well to mix the solution.
3. Using a wash bottle rinse the sides of the flask.
4. Titrate it with N/50 Na2S2O3 solution from the burette till the solution becomes straw yellow.
5. Add 2 ml of the starch solution. The solution will turn blue.
6. Continue titration with Na2S2O3 till the blue colour disappears.
7. Note the final reading and repeat to get concordant readings.

OBSERVATIONS:
Normality of the Na2S2O3 solution = N/50
Observation Table
S. N. Volume of the solution Burette Readings Volume of the
taken in the titration flask titrant used (final-
(ml) Initial Final Reading initial reading) (ml)
Reading
1
2
3
CALCULATIONS:
Sample = Na2S2O3 solution
N1V1 = N2V2
1
N1 × 100 =  V2
50
V2
N1 
5000
Total residual Cl2 = N1 × Equivalent Weight of Cl2 = N1 × 35.5 g L-1 = N1 × 35.5 × 1000 mg L-1
RESULT: The amount of residual Cl2 in water sample = …….mg L-1.
PRECAUTIONS:
1. Chlorine vapours are harmful so the solution should not be sucked in to the pipette with mouth.
2. The titration should be completed rapidly in order to avoid atmospheric oxidation of iodide.
3. First disappearance of blue colour should be taken as end point.
Instruction to the Lab Assistant
Provide the following
1. N/50 Na2S2O3 solution: Dissolve 4.96 g of Na2S2O3 in distilled water and make up the volume to 1 litre.
Standardize with N/50 K2Cr2O7.
2. Glacial Acetic Acid
3. KI (Solid)
4. Starch solution
Viva Questions:
1. Why Cl2 is added to water?
2. What are the disadvantages of excess Cl2 present in water?
3. What is the action of Cl2 on pathogens?
4. Why and how pH is adjusted between 3 and 4?
Ans. pH is adjusted between 3 and 4 using acetic acid. At neutral pH, some combined chlorine residuals do
not liberate I2 by reacting with KI.
 EXPERIMENT NO. 7

AIM: Determination of Ammonia in Water.

THEORY: Ammonia is found as ammonium hydroxide in solution and reacts with sulfuric acid according to
the following equation.
H2SO4  2NH 4OH  (NH 4 )2 SO4  2H2 O
REAGENT: Standard solution of H2SO4
METHOD:
1. Rinse and fill the burette with N/50 H2SO4.
2. Pipette out 25mL of the water solution in titration flask.
3. Add 2 drops of methyl red indicator and mix well.
4. Run the acid solution into the flask till the colour changes from yellow to red.
5. Repeat to get three concordant readings.

OBSERVATIONS:
Normality of the acid solution = N/50
Volume of mixture taken for each titration =25mL
Observation Table

S. N. Volume of the water sample Burette Readings Volume of the

taken in the titration flask titrant used (final-
(ml) Initial Final Reading initial reading) (ml)
Reading
1
2
3

CALCULATIONS:
Acid = Water solution
N1V1  N2 V2
1
V1  N 2  25
50
1 V
N2   1
50 25
Strength = N2 ×17 g L-1 = N2 ×17×1000 mg L-1
RESULT: The strength of ammonia in water sample is……. mg L-1.
 EXPERIMENT NO. 8

AIM: Determination of free CO2 in given sample of water.

APPARATUS: Burette, Pipette, Conical flask, Beakers etc.
CHEM CALS: N/50 Na2CO3 or N/50 NaOH), phenolphthalein
THEORY: Free CO2 is determined by titrating the sample water with N/50 NaOH or N/50 Na2CO3 solution
using phenolphthalein as indicator.
The CO2 reacts with NaOH or Na2CO3 to form HCO3 .

CO 2 + H 2 O  HCO 3
CO 2 + CO 32- +H 2 O  2HCO 3
The end point is appearance of pink colour.
PROCEDURE:
1. Rinse and fill the burette with N/50 Na2CO3 solution
2. Pipette out 100 ml of water sample in a conical flask (there should be no air bubbles). Add a few drops of
phenolphthalein.
3. Titrate the solution With N/50 Na2CO3 from the burette, till the pink colour persists for at least 30 seconds.
4. Note the reading and repeat to get three concordant readings.
OBSERVATIONS:
Volume of water sample taken for each titration = 100 ml
Normality of Na2CO3 used = N/50
Observation Table:

S. N. Volume of the water sample Burette Readings Volume of the

taken in the titration flask titrant used (final-
(ml) Initial Final Reading initial reading) (ml)
Reading
1
2
3

CALCULATIONS:
Water sample = Na2CO3
N1V1  N2 V2
1
N1 100=  V2
50
1
N1 =  V2
5000
Strength of CO2 = N1 × Eq. Wt.
 V2 V
Strength of CO 2 =  22g L1 = Strength of CO 2 = 2  22  1000 mg L1
5000 5000
RESULT: The amount of free CO2 present in water sample is……..ppm.
PRECAUTIONS:
1. The reaction mixture should be stirred very gently during titration.
2. Same amount of indicator should be added while taking different readings.
Instructions to the Lab Assistant
Provide the following
1. N/50 Na2CO3 solution: Dissolve 1.06 g of Na2CO3 in distilled water and make up the volume to 1
litre.
2. Phenolphthalein.
Viva Questions:
1. What is the source of acidity in water?
2. How the acidity of water is determined?
3. What are the disadvantages of acidic water?
4. What are the disadvantages of dissolved CO2 present in water?
5. How can you determine the amount of dissolved CO2 in water?
 EXPERIMENT NO 9

AIM: Determination of pH of some soil samples.

APPARATUS: pH meter or potentiometer, glass electrode, thermometer, beaker, stirrer
THEORY: Soil acts as an electrolyte and hence its pH can be measured. The pH is measured to determine the
degree of acidity or alkalinity in the soil sample. The pH of soil is an important physico-chemical characteristic
because it influences:
• The suitability of a soil for crop production.
• The availability of nutrients in the soil.
• The microbial activity in the soil.
• The lime and the gypsum requirement.
• The soil's physical properties like structure, permeability etc.
For determining the pH of a solution different methods employed. The most widely used method is by using
pH meter. The value of pH, the logarithm of the reciprocal of the hydrogen ion concentration in solution, is
determined by measuring the difference in potential between two electrodes immersed in a sample solution.
The method is based on the potential difference between an electrode pair consisting of a glass electrode
sensitive to the difference in the hydrogen ion activity in the sample solution and the internal filling solution,
and a reference electrode, which is-supposed to have a constant potential independent of the immersing solution.
These days, combined electrodes are used in which the reference electrode is combined with the glass electrode.
The measured potential difference is compared with the potential obtained when both electrodes are immersed
in a solution or buffer with known pH or hydrogen ion concentration. The pH is defined by the formula:

F
pH(sample)  pH(reference)  E (sample)  E (reference)
RTln10
Where, E represent the electrode potentials, R is the universal gas constant, T the absolute temperature and F is
the Faraday constant.

pH interpretation :

S. N. pH Interpretation
1 Below 6.5 Acidic- Requires liming
2 6.5-8.7 Normal- No treatment, optimum for most of the crops
3 Above 8.7 Alkaline-Requires reclamation with gypsum and by growing the salt toleration
crops

PROCEDURE:
1. Soil Sample:
(i) Take the soil sample with the help of clean spatula and crush it if the lumps are there.
 (ii) Take around 50 g of the sample in a beaker and add 100 mL of distilled water. Stir at least 4
times over a 30 minutes period or continuously shaking for 10 min on mechanical shaker. This
time is allowed for the soil and water to reach equilibrium.
(iii) Shake the soil suspension and filter it.

2. Calibration: Calibrate the electrode as per the following procedure:

(i) Connect the pH meter to the power supply. Switch on the instrument.
(ii) Take a standard buffer solution of pH 7 in 100 mL beaker. Note the temperature of the buffer
solution.
(iii) Set the temperature control of the pH meter to the temperature of the buffer solution.
(iv) Remove the combination electrode from the storage solution, wash it with distilled water and blot
dry with soft tissue paper.
(v) Connect the combination electrode to the input socket and dip the electrode in 7 pH buffer solution.
(vi) Set the Function Selector Switch to pH position and adjust with calibrate control till the digital
display shows the pH 7.
(vii) Now move the function switch to ‘Stand by' position.
(viii) Remove the electrode from the buffer solution and wash it with distilled water. Dry with tissue paper.
(ix) Dip the electrode in the buffer solution having pH 4.
(x) Set the "temperature" to the temperature of the solution.
(xi) Set the Function Selector Switch to pH position and adjust with 'Slope correction" control till the
digital display shows the pH 4.

3. pH measurements:
(i) Connect the combination electrode to the input socket after washing it with distilled water.
(ii) Dip the combined electrode in the soil solution under test.
(iii) Set the Temperature knob to the temperature of the solution.
(iv) Set the Function Selector Switch to pH position.
(v) Note the pH of the solution.
(vi) Repeat the same procedure for other soil samples.
OBSERVATIONS: Soil sample
S.N. Sample pH
1
2
3
4
RESULT: The pH of the given soil samples are……..
PRECAUTIONS:
1. Immediately after testing, the electrodes should be washed off with a gentle stream of distilled H2O.
2. For storage, after cleaning, the electrodes are suspended in distilled water and the system is protected
against evaporation. The drying out of the electrodes should be avoided.
 EXPERIMENT NO 10
AIM: Synthesis of dibenzalpropanone (Base catalysed Aldol condensation by Green methodology).
REQUIREMENTS:
Chemicals: Acetone 1ml (0.83 g), benzaldehyde 3.8 ml (3.9 g), NaOH (30 ml) of 10% solution, Ethanol
(15 ml)
Apparatus: Conical flask, Filtration set up.
THEORY: Dibenzalpropanone is prepared by base catalysed Aldol condensation method.

Mechanism of this reaction is given below:

PROCEDURE:

(i) Take 3.8 mL benzaldehyde, 1 mL acetone; 15 mL alcohol in a conical flask. Fit a cork over the
mouth of this conical flask and shake for 2 minutes.
(ii) Add 30 mL of 10% sodium hydroxide solution.
(iii) Shake vigorously for 10 minutes with simultaneous pressure release.
 (iv) Cool the reaction mixture in ice.
(v) Filter the pale yellow solid through filter paper.
(vi) Wash with water, dry, collect, weigh, and recrystallize from ethanol.

RESULT: Yield = 3.0 g (90%)

SIGNIFICANCE:

In this synthesis method non-toxic reagents are used. Hazardous organic solvents are avoided so it is a green
methodology.