EELabManual Final

INTRODUCTION
Water is vital for life. Not only do we need water to drink, to grow food and to wash, but it is
also important for many of the pleasant recreational aspects of life. The impurities which
water dissolves or picks up as suspended matter, may sometimes make it more useful and
portable for public used and especially for drinking, and sometimes it may render it harmful
and unfit. For example, certain minerals such as iron, calcium, magnesium, fluorine etc. in
small quantities may be useful and good for health of the people, because human beings need
a certain amount of these elements in their bodies. But when these materials and others are
dissolved in large amounts or in certain combinations, the water may become unfit for
municipal, industrial and other uses. Similarly, sometimes the water may contain harmful
bacteria, the presence of which may cause diseases such as cholera, typhoid, dysentery,
gastro-enteritis, infectious hepatitis, etc.
To ensure safety to public health, economy and utility in industries and other uses, it,
therefore, becomes imperative upon the planners and designers of the public water supply
schemes, to thoroughly check, analyse, and treat the raw available water to safe and
permissible limits, before supplying to public. This is truer and must be strictly adhered to,
when water is supplied for municipal uses, such as drinking, bathing, washing etc. When the
water is required solely for industries and for irrigation, the quality standards may vary
depending upon the requirement of each industry or farming and irrigation. The quality
requirements for certain industries are so great that these industries have to develop their own
supplies.
CHARACTERISTICS OF WATER
The raw or treated water can be checked and analyzed by studying and testing their physical,
chemical and biological characteristics.
Physical Characteristics: Turbidity, Colour, Taste, Odour, Temperature, Electrical

conductivity etc.
Chemical Characteristics: Solids, pH, Hardness, Alkalinity, Chloride, Metals, Dissolved

gases, Biochemical oxygen demand, Chemical oxygen demand, Fats and oils and grease,
Volatile organic compounds, Toxic metal and non-metal ions, etc.
1 Environmental Engineering Laboratory Manual – NIT Calicut, Kerala

Biological Characteristics: Bacteria, Protozoa, Viruses, Pathogens, etc.
TYPES OF WATER ANALYSIS
Gravimetric Analysis: Gravimetric analysis describes a set of methods in analytical

chemistry for the quantitative determination of an analyte based on the mass of a solid. A
simple example is the measurement of solids in a water sample.
In most cases, the analyte must first be converted to a solid by precipitation with an
appropriate reagent. The precipitate can then be collected by filtration, washed, dried to
remove traces of moisture from the solution, and weighed. The amount of analyte in the
original sample can then be calculated from the mass of the precipitate and its chemical
composition.
In other cases, it may be easier to remove the analyte by evaporation. The analyte might be
collected—perhaps in a cryogenic trap or on some absorbent material such as activated
carbon—and measured directly. Or, the sample can be weighed before and after it is dried;
the difference between the two masses gives the mass of analyte lost. This is especially useful
in determining the water content of complex materials such as foodstuffs.
Volumetric Analysis (Titrimetry): Volumetric analysis is a widely-used quantitative

analytical method. As the name implies, this method involves the measurement of volume of
a solution of known concentration which is used to determine the concentration of the
analyte.
Colourimetric Analysis: It is a method of determining the concentration of a chemical

element or chemical compound in a solution with the aid of a colour reagent. It is applicable
to both organic compounds and inorganic compounds and may be used with or without
an enzymatic stage. The method is widely used in medical laboratories and for industrial
purposes, e.g. the analysis of water samples in connection with industrial water treatment.
A colourimeter is a device used to test the concentration of a solution by measuring its

absorbance of a specific wavelength of light. To use the colourimeter,
different solutions must be made, including a control or reference of known concentration.
With a visual colourimeter, for example the Duboscq colourimeter, the length of the light
path through the solutions can be varied while filtered light transmitted through them is

compared for a visual match. The concentration times the path length is taken to be equal
when the colours match, so the concentration of the unknown can be determined by simple
proportions. Nessler tubes work on the same principle.
There are also electronic automated colourimeters; before these machines are used, they must
be calibrated with a cuvette containing the control solution. The concentration of a sample
can be calculated from the intensity of light before and after it passes through the sample by
using the Beer–Lambert law. Photoelectric analyzers came to dominate in the 1960s.
The colour or wavelength of the filter chosen for the colourimeter is extremely important, as
the wavelength of light that is transmitted by the colourimeter has to be the same as that
absorbed by the substance being measured. For example, the filter on a colourimeter might be
set to red if the liquid is blue.
INSTRUCTIONS
1. Discuss the results after each experiment based on the following points:
 Limit prescribed for that constituent in drinking water standards.
 The suitability of the sample for drinking purpose with respect to that particular
constituent.
 Other uses
2. References for writing the discussion after each experiment are:
 “Standard Methods for the Examination of Water and Waste Water”, American
Public Health Association, 1015, 15th Street, N.W., Washington D.C., 2005.
 “Chemistry for Environmental Engineers”, Sawyer and McCarty, Tata Mc-Graw
Hill.
 “Wastewater Engineering Treatment, Disposal and Reuse”, Metcalf and Eddy,
Tata McGraw Hill.
 “Water and Wastewater Engineering, Design principles and practice”, M L Davis,
McGraw Hill, New York.
 “Environmental Engineering, Vol. 1” S. K. Garg, Khanna Publications, New
Delhi.
 “International Standards for Drinking Water” — World Health Organisation.
 “IS 10500-2012” Bureau of Indian Standards, New Delhi.

 “The Environment (Protection) Rules -1986” Ministry of Environment and
Forests, New Delhi.
 Other relevant standards.
DOs AND DON’Ts IN THE LABORATORY
 Do thoroughly clean the glassware before and after use.

 Do handle the glassware carefully.
 Do not handle chemicals with bare hands.
 Do not blow out the last drop from the pipette. When the liquid has drained out
completely, touch the tip of the pipette to the inner surface of the vessel.
 Do not add water to acids. Do always add acid to water.
 Do use large volumes of water, when a person is splashed with acid to prevent
serious burns.
 Do weigh the articles in a balance only at room temperature.
 Do use different pipette for different reagents.
 Do not pipette out acids and other toxic reagents by mouth.
 Do read the level of the curve (meniscus), in all volumetric glassware, with the eye
at approximately the same level as the curve of solution.

DETERMINATION OF pH
INTRODUCTION
The term pH refers to the measure of hydrogen ion concentration in the solution and defined
as the negative log of H+ ions concentration in the water and wastewater. The values of pH 0
to a little less than 7 are termed as acidic and the values of pH a little above 7 to 14 are
termed as basic. When the concentration of H+ and OH- ions are equal, then it is termed as
neutral pH.
ENVIRONMENTAL SIGNIFICANCE
 Lower value of pH below 4 will produce sour taste and higher value above 8.5 a bitter
taste.
 Higher values of pH hasten the scale formation in water heating apparatus and also
reduce the germicidal potential of chlorine.
 High pH induces the formation of trihalomethanes, which can cause cancer in human
beings.
 pH below 6.5 starts corrosion of pipes.
 It is used in the calculation of carbonate, bicarbonate, stability index and acid base
equilibrium.
 Determination of pH is important in biological treatment of wastewater. In anaerobic
treatment, if the pH goes below 6.5 due to excess accumulation of acids, the process is
severely affected. Shifting of pH beyond 5 to 10 upsets the aerobic treatment of the
wastewater. In these circumstances, the pH can be adjusted by the addition of suitable
acids and alkali to optimize the treatment of wastewater.
 pH value or range is of immense importance for any chemical reaction. A chemical
shall be highly effective at a particular pH. Chemical coagulation, disinfection, water
softening and corrosion control are governed by pH adjustment.
 Dewatering of sludge, oxidation of cyanides and reduction of Cr6+ to Cr3+ also need a
favourable pH range.
AIM
To determine the pH value of the given water sample.

PRINCIPLE
pH value of water indicates the hydrogen ion concentration in water and concept of pH was
put forward by Sorenson (1909). pH is expressed as the logarithm of the reciprocal of the
hydrogen ion concentration in moles/litre at a given temperature. The pH scale extends from
0 (very acidic) to 14 (very alkaline) with 7 corresponding to exact neutrality at 25°C. pH can
be measured colourimetrically or electrometrically. Colourimetric method is used only for
rough estimation. It can be done either by using universal indicator or by using pH paper. The
hydrogen electrode is the absolute standard for the measurement of pH. They range from
portable battery operated units to highly precise instruments. But glass electrode is less
subjected to interferences and is used in combination with a calomel reference electrode. This
system is based on the fact that a change of 1 pH unit produces an electric charge of 59.1 mV
at 25°C.
APPARATUS
o pH meter with electrode

o Beaker
o Thermometer
o Colour comparator with discs
o Cuvettes
REAGENTS
 Buffer solutions
 pH paper
 Universal indicator
PROCEDURE
(a) Using Universal Indicator
1. 10 mL of sample is taken in a cuvette.

2. Another 10 mL sample is taken in another cuvette and 0.2 mL of universal indicator is
added. The cuvettes are placed in the holes provided for it.
3. A colour disc corresponding to this indicator is inserted into the comparator and the
disc rotated such that the 2 circles indicate identical colours.
4. The reading is noted.
5. The procedure can be repeated using an indicator whose range is near the value
obtained.
6. The exact pH is obtained.
(b) Using pH Papers
1. Dip the pH paper in the sample.

2. Compare the colour with that of the colour given on the wrapper of the pH paper
book.
3. Note down the pH of the sample along with its temperature.
(c) Using pH Meter
1. Follow the manufacturer’s operating instructions.

2. Calibrate instrument.
3. A solution whose pH is to be found is taken in a beaker and the temperature knob is
adjusted such that the temperature of solution is same as that in dial.
4. The electrode is washed with distilled water and rinsed with the solution and then it is
dipped in the solution.
5. The reading on the dial indicates the pH of the solution
OBSERVATIONS
Temperature of the sample = .................................... oC
pH
Sample No.
pH Paper pH Meter Universal Indicator

RESULT
The pH of the given sample is = ..................................................
QUESTIONS
1. A decrease in pH of 1 unit represents how much of an increase in hydrogen ion

concentration?
2. Discuss the relationship between (a) pH and hydrogen ion concentration (b) pH and
hydroxide ion concentration?
3. What is a buffer solution? Give examples.
4. What is the pH of (a) 2N HCl (b) 0.02 N NaOH (c) a solution containing 8 mg/L HCl
and 4 mg/L NaOH
5. Explain the working principle of pH meter.
6. Explain the effect of temperature on pH.

DETERMINATION OF TURBIDITY
INTRODUCTION
Clarity of water is important in producing products destined for human consumption and in
many manufacturing operations. Beverage producers, food processors, and potable water
treatment plants drawing from a surface water source commonly rely on fluid-particle
separation processes such as sedimentation and filtration to increase clarity and insure an
acceptable product. The clarity of a natural body of water is an important determinant of its
condition and productivity.
Turbidity in water is caused by suspended and colloidal matter such as clay, silt, finely
divided organic and inorganic matter, and plankton and other microscopic organisms.
Turbidity is an expression of the optical property that causes light to be scattered and
absorbed rather than transmitted with no change in direction or flux level through the sample.
Correlation of turbidity with the weight or particle number concentration of suspended matter
is difficult because the size, shape, and refractive index of the particles affect the light-
scattering properties of the suspension. When present in significant concentrations, particles
consisting of light-absorbing materials such as activated carbon cause a negative interference.
In low concentrations these particles tend to have a positive influence because they contribute
to turbidity. The presence of dissolved, colour-causing substances that absorb light may cause
a negative interference. Some commercial instruments may have the capability of either
correcting for a slight colour interference or optically blanking out the colour effect.
 Turbidity is objectionable because of (a) aesthetic considerations and (b) engineering

considerations.
 When the turbid water in a small, transparent container such as drinking glass is held
up to the light, an aesthetically displeasing opaqueness or milky colouration is
apparent.
 The colloidal material which exerts turbidity provides adsorption sites for chemicals
they may be harmful or cause undesirable tastes and odours, and for biological
organism that may be harmful.

 Disinfection of turbid water is difficult because the adsorptive characteristics of some
colloids and because the solids may partially shield organisms from disinfectant.
 In natural water bodies, turbidity may impart a brown or other colour to water and
may interfere with light penetration and photosynthetic reaction in streams and lakes.
 Turbidity increases the load on slow sand filters. The filter may go out of operation, if
excess turbidity exists.
 Turbidity measures are of particular importance in the field of water supply. They
have limited use in the field of domestic and industrial waste treatment.
AIM
To determine the turbidity of the given water sample.
PRINCIPLE
This method is based on a comparison of the intensity of light scattered by the sample under
defined conditions with the intensity of light scattered by a standard reference suspension
under the same conditions. Higher the intensity of scattered light, the higher the turbidity.
Formazin polymer is used as the primary standard reference suspension. The turbidity of a
specified concentration of formazin suspension is defined as 4000 NTU.
APPARATUS
o Nephelometric turbidity meter

o Sample vials
o Standard flasks
o Measuring cylinder
REAGENTS
 Hexamethylene tetramine
 Hydrazine sulphate
 Distilled water
REAGENTS PREPARATION
Stock Primary Standard Formazin Suspension: Dissolve 1 g hydrazine sulfate,

(NH2)2⋅H2SO4, in distilled water and dilute to 100 mL in a volumetric flask. Dissolve 10 g
hexamethylenetetramine, (CH2)6N4, in distilled water and dilute to 100 mL in a volumetric
flask. In a flask, mix 5 mL each of the above solution and keep it for 24 h at 25 ± 3°C. This
results in a 4000 NTU suspension. Transfer stock suspension to an amber glass or other UV-
light-blocking bottle for storage. Make dilutions from this stock suspension. The stock
suspension is stable for up to 1 year when properly stored.
Dilute Turbidity Suspensions: Dilute 4000 NTU primary standard suspension with high-
quality distilled water. Prepare immediately before use and discard after use.
PROCEDURE
1. Switch on the instrument.

2. Calibrate the instrument.
3. Thoroughly shake the sample.
4. Wait until air bubbles disappear and pour the sample into the nephelometer tube.
5. Read the turbidity directly from the instrument.
OBSERVATIONS
No. Sample No. Turbidity (NTU)
RESULT
The turbidity of the given sample is = .................................................. NTU
QUESTIONS
1. Where do you find the adverse effects of turbidity in environmental engineering?

Mention two instances.
2. Discuss the nature of materials causing turbidity in
(a) River water during flash flood
(b) Polluted river water
(c) Domestic wastewater
3. What is the standard unit of turbidity?
4. Discuss why turbidity in general cannot be correlated with the weight concentration of
suspended matter in water samples.
5. Explain the working principles of Nephelometric and Jackson turbidity meter.
6. Why is silica no longer the preferred material for turbidity standards?

DETERMINATION OF SOLIDS
INTRODUCTION
The term “solids” is generally used when referring to any material suspended or dissolved in
water or wastewater that can be physically isolated either through filtration or through
evaporation. In stream water, dissolved solids consist of calcium, chlorides, nitrate,
phosphorus, iron, sulfur, and other ions particles that will pass through a filter with pores of
around 2 microns (0.002 cm) in size. Suspended solids include silt and clay particles,
plankton, algae, fine organic debris, and other particulate matter. These are particles that will
not pass through a 2-micron filter. Solids can also be classified as either filterable or non-
filterable. Filterable solids may either be settleable or non-settleable.
 Total solids measurements can be useful as an indicator of the effects of runoff from
construction, agricultural practices, logging activities, sewage treatment plant
discharges and other sources.
 Although the sewage normally contains 99.9% of water and 0.1% of solids, it is the
solids that have the nuisance value.
 Solids analyses are important in the control of biological and physical wastewater
treatment processes and for assessing compliance with regulatory wastewater effluent
limitations.
 If the solids in the wastewater are mostly organic, the impact on a treatment plant is
greater than if the solids are mostly inorganic.
 Dissolved minerals, gases and organic constituents may produce aesthetically
displeasing colour, taste and odour.
 Water with higher solids content often has a laxative and sometimes the reverse effect
upon people whose bodies are not adjusted to them.
 High concentration of dissolved solids, about 3000 mg/L, may also produce distress in
livestock.
 Suspended material is aesthetically displeasing and provides adsorption sites for
chemical and biological agents.

 Some dissolved organic chemicals may deplete the dissolved oxygen in receiving
water and some may be inert to biological oxidation, yet others have been identified
as carcinogens.
 Volatile solids test is normally applied to sludges. It is indispensable in the design and
operation of sludge digester, vacuum filter and incineration plants.
AIM
The aim of the experiments is to determine the following types of solids in the given
sample(s):
(a) Total solids

(b) Total (inorganic) fixed solids
(c) Total volatile (organic) solids
(d) Total dissolved solids
(e) Dissolved fixed (inorganic) solids
(f) Dissolved volatile (organic) solids
(g) Total suspended solids
(h) Suspended fixed (inorganic) solids
(i) Suspended volatile (organic) solids
(j) Settleable solids
PRINCIPLE
‘Total solids’ is the term applied to the material left in the vessel after evaporation of a
sample of water/waste water and its subsequent drying in an oven at a definite temperature.
Total solids include “total suspended solids” the portion of total solids retained by a filter and
“total dissolved solids” the portion that passes through the filter. Fixed solid is the residue
remaining after ignition for 1 hour at 550°C. The solid portion that is volatilised during
ignition is called volatile solids. It will be mostly organic matter. Waters that are low in
organic matter and total mineral content and are intended for human consumption may be
examined under 103–105°C or 179–181°C. But water containing considerable organic matter
or those with pH over 9.0 should be dried at 179–181°C. In any case, the report should
indicate the drying temperature.

The sample is filtered and the filtrate evaporate in a weighed dish on a steam bath, the residue
left after evaporation is dried to constant weight in an oven at either 103–105°C or 179–
181°C. The increase in weight over that of the empty dish represents total dissolved solids
and includes all materials, liquid or solid, in solution or otherwise, which passes through the
filter and not volatilised during the drying process.
The difference between the total solids and the total dissolved solids will give the total
suspended solids. The dishes with the residue retained after completion of the tests for total
solids and total dissolved solids are subjected to heat for 1 hour in a muffle furnace held at
550°C. The increase in weight over that of the ignited empty vessel represents fixed solids in
each instance.
The difference between the total dissolved/total suspended solids and the corresponding fixed
solids will give volatile solids in each instance. All the quantities should be expressed in
mg/L. Settleable matter in surface and saline waters as well as domestic and industrial wastes
may be determined and reported on a volume basis as millilitre per litre.
APPARATUS
o Porcelain evaporating dishes of 150–200 mL capacity

o Steam bath
o Drying oven
o Desiccators
o Analytical balance or monopan balance
o Filter paper (preferably of glass fibre)
o Electric muffle furnace
o Imhoff cone
PROCEDURE
(a) Total Solids
1. Ignite the clean evaporating dishes in the muffle furnace for 30 minutes at 550°C and
cool in a desiccator.
2. Note down the empty weight of the dish (W1).

3. Pour a measured portion (50 to 100 mL) of the well-mixed sample into the dish and
evaporate the contents by placing the dish on a steam bath.
4. Transfer the dish to an oven maintained at either 103–105°C or 179–181°C and dry it
for 1 hour.
5. Allow the dish to cool briefly in air before placing it, while still warm in a desiccator
to complete cooling in a dry atmosphere.
6. Weigh the dish as soon as it has completely cooled (W2).
7. Weight of residue = (W2 – W1) mg. W2 and W1 should be expressed in mg.
(b) Total Fixed Solids
1. Keep the same dish used for determining total residue in a muffle furnace for 1 hour
at 550°C.
2. Allow the dish to partially cool in air until most of the heat has dissipated, then
transfer to a desiccators for final cooling in a dry atmosphere.
3. Weigh the dish as soon as it has cooled (W3).
4. Weight of total fixed residue = (W3 – W1) mg. W3 and W1 should be expressed in mg.
(c) Total Dissolved Solids
1. Filter a measured portion of the mixed sample (50 or 100 mL) through a filter paper
and collect the filtrate in a previously prepared and weighed evaporating dish.
2. Repeat the steps 3 to 6 outlined in total solids procedure.
3. Weight of dissolved solids = (W5 – W4) mg.
W4 = Weight of empty evaporating dish in mg.
W5 = Weight of empty evaporating dish in mg + Residue left after evaporating the
filtrate in mg.
(d) Total Suspended Solids = Total solids – Total dissolved solids.
(e) Total Volatile Solids = Total solids – Total fixed solids.
(f) Fixed Dissolved Solids
1. Keep the same evaporating dish used in determining total dissolved solids in a muffle
furnace for 1 hour at 550°C.
2. Repeat the steps 2 and 3 outlined in total fixed solids procedure.
3. Weight of fixed dissolved solids = (W6 – W4) mg.
W6 = Weight of empty evaporating dish + Fixed solids left after ignition at 550°C.
(g) Volatile Dissolved Solids = Total dissolved solids – Fixed dissolved solids.
(h) Fixed Suspended Solids = Total fixed solids – Fixed dissolved solids.
(i) Volatile Suspended Solids = Total volatile solids – Volatile dissolved solids.
(j) Settleable Solids by Volume
1. Fill an imhoff cone to the litre mark with a thoroughly mixed sample.
2. Settle for 45 minutes.
3. Gently stir the sides of the cone with a rod or by spinning.
4. Settle 15 minutes longer.
5. Record the volume of settleable matter in the cone as mL/L.
CALCULATION

OBSERVATIONS
Total Solids
Item Sample 1 Sample 2 Sample 3
Volume of Sample (mL)
Empty Wt. of Dish (W1; mg)
Wt. of Dish + Total Solids ( W2; mg)
Total Solids (mg/L)
Total Fixed Solids
Wt. of Dish + Total Fixed Solids ( W3; mg)
Total Fixed Solids (mg/L)
Total Dissolved Solids
Wt. of Dish + Total Dissolved Solids ( W5; mg)
Total Dissolved Solids (mg/L)

Fixed Dissolved Solids
Wt. of Dish + Fixed Dissolved Solids ( W6; mg)
Fixed Dissolved Solids (mg/L)
Settleable Solids
Volume of sample = ................................ mL
Settled solids =.................................. mL
Settleable solids = .............................. mL/L
RESULT
Total Solids (mg/L)
Total Fixed Solids (mg/L)
Total Dissolved Solids (mg/L)
Total Suspended Solids (mg/L)
Total Volatile Solids (mg/L)
Fixed Dissolved Solids (mg/L)
Volatile Dissolved Solids (mg/L)
Fixed Suspended Solids (mg/L)
Volatile Suspended Solids (mg/L)
Settleable Solids (mL/L)

QUESTIONS
1. What is the application of determination of settleable solids?

2. Explain the significance of determination of total solids in sanitary engineering.
3. How will the volatile solids affect the strength of sewage? Why?
4. Why do you determine the fixed solids by igniting at 550°C?
5. What significant information is furnished by the determination of volatile solids?
6. What is sludge volume index?
7. With the help of a pictorial representation, explain the various components of total
solids. Explain their properties and inter-relationships.
8. The following test results were obtained for a wastewater sample. All the tests were
performed using a sample size of 50 mL. Determine the concentrations of total solids,
total volatile solids, suspended solids, volatile suspended solids, total dissolved solids
and total volatile dissolved solids.
Tare mass of evaporating dish after drying at 105oC: 53.5433g
Mass of evaporating dish plus residue after evaporating at 105oC: 53.5794g
Mass of evaporating dish plus residue after ignition at 550oC: 53.5625g
Tare mass of Whatman GF/C filter paper after drying at 105oC: 1.5433 g
Mass of Whatman GF/C filter paper plus residue after evaporating at 105oC: 1.5554g
Mass of Whatman GF/C filter paper plus residue after ignition at 550oC: 1.5476g

DETERMINATION OF CHLORIDES
INTRODUCTION
Chloride occurs in all natural water in widely varying concentrations. The chloride content
normally increases as the mineral content increases. Upland and mountain supplies usually
are quite low in chloride, whereas river and groundwater usually have a considerable amount.
Sea and ocean water represent the residues resulting from partial evaporation of natural water
that flow into them, and chloride levels are very high.
Chlorides are generally present in water in the form of sodium chloride and may be due to:
leaching of marine sedimentary deposits, pollution from sea water, brine or industrial and
domestic wastes, etc. The presence of high quantity of chloride in river or stream water may
indicate the pollution of water due to sewage and other human or industrial wastes. The
chloride concentrations of raw water, being used for public supplies, should therefore
regularly be tested, so as to immediately detect any sudden increase in its chloride content
and the possibility of any organic pollution of water.
 Chloride in reasonable amount is not harmful to humans. Chlorides associated with

NaCl exert salty taste when its concentration is more than 250 mg/L and is
objectionable to many people. Therefore, the chlorides are limited to 250 mg/L in
water intended for public supply.
 In many areas of the world where water supplies are scare, sources containing as
much as 2000 mg/L are used for domestic purposes without the development of
adverse effects, once the human system becomes adapted to the water.
 It can also corrode concrete by extracting calcium in the form of calcide.
 Magnesium chloride in water generates HCl after heating, which is also highly
corrosive and create problems in boilers.
 Chloride determination in natural water is useful in the selection of water supplies for
human use.
 The chloride content of water used for irrigation of agricultural crops is generally
controlled along with the total salinity of water. Evapotranspiration tends to increase
the chloride and salinity at the root zone of irrigated plants, making it difficult for

crops to take up water due to the osmotic pressure differences between the water
outside the plants and within the plant cells. For this reason, chloride and total salinity
concentrations at or below the drinking water standards are normally specified for
water to irrigate salt-sensitive crops.
 Chloride determination is used to determine the type of desalting apparatus to be used.
 Chloride is used to some extent as a tracer in environmental engineering practice.
 Chloride determination is used to control pumping of groundwater from the locations
where sea water intrusion is a problem.
 Chlorides interfere in the determination of chemical oxygen demand.
AIM
To determine the amount of chloride (in the form of Cl–) present in the given water sample.
PRINCIPLE
The Mohr (Argentometric) method is used to determine the chloride concentration present in
the water sample. The water sample is titrated with standard silver nitrate solution (0.0282 or
0.0141 N), in which silver chloride is precipitated at first (Eq. 1). Since silver chloride is a
white precipitate, the end point cannot be detected visually, unless an indicator capable of
demonstrating the presence of excess silver ions is present. The indicator normally used is
potassium chromate, which supplies chromate ions. As the concentration of chloride ions
approaches extinction, the silver ion concentration increases to a level at which the solubility
product of silver chromate is exceeded and it begins to form a reddish brown precipitate as in
Eq. 2. This is taken as evidence that all the chloride has been precipitated. Since an excess of
silver ions is needed to produce a visible amount of silver chromate, the indicator error or
blank must be determined and subtracted from all titrations.
(1)
(2)
APPARATUS
o Burette
o Pipettes
o Erlenmeyer flasks
REAGENTS
 Chloride free distilled water.

 Standard silver nitrate solution
 Potassium chromate indicator.
 Acid or alkali for adjusting pH.
Standard Silver Nitrate (0.0141 N): Dissolve 2.395 g of silver nitrate in distilled water and
make up to 1000 mL in a volumetric flask. Standardize it against 0.0141 N NaCl solution.
Store it in an amber coloured bottle.
Potassium Chromate Indicator: Dissolve 50 g K2CrO4 in a little distilled water. Add

AgNO3 solution until a definite red precipitate is formed. Let stand 12 h, filter, and dilute to 1
L with distilled water.
PROCEDURE
1. Take 20 mL of the given sample (V) in a conical flask. If the sample is coloured add 3
mL of aluminium hydroxide, shake well; allow to settle, filter, wash and collect
filtrate. If sulphide, sulphite, or thiosulphate is present, add 1 mL H2O2 and stir for 1
min.
2. Sample is brought to pH 7–8 by adding acid or alkali as required.
3. Add 1mL of indicator (Potassium chromate) to get light yellow colour.
4. Titrate the solution against standard silver nitrate solution until a reddish brown
precipitate is obtained.
5. Note down the volume of silver nitrate added (V1).
6. Repeat the procedure for blank and note down the volume (V2).
CALCULATION

OBSERVATIONS
Burette solution: Silver Nitrate

Indicator: Potassium Chromate
End Point: Yellow to Reddish Brown
Titration 1 (Sample)
Volume of Sample Burette Reading (mL) Volume of AgNO3
No.
(mL) Initial Final V1 (mL)
Titration 2 (Distilled water)

Volume of Sample Burette Reading (mL) Volume of AgNO3
No.
(mL) Initial Final V2 (mL)
RESULT
The amount of chloride present in the given sample is = .................................................. mg/L
QUESTIONS
1. Explain the need for blank correction.

2. Why must the sample pH be neither high nor low?

3. Would the analytical result by Mohr’s method for chlorides be higher, lower, or the
same as the true value if an excess indicator were accidentally added to the sample?
Why?
4. Why water has a lower content of salt than sewage?
5. Why silver nitrate solution is stored in amber coloured bottle?

DETERMINATION OF ALKALINITY
INTRODUCTION
Alkalinity of water is its acid-neutralizing capacity. It is the sum of all the titratable bases.
The measured value may vary significantly with the end-point pH used. Alkalinity is a
measure of an aggregate property of water and can be interpreted in terms of specific
substances only when the chemical composition of the sample is known.
Alkalinity is significant in many uses and treatments of natural waters and wastewaters.
Because the alkalinity of many surface waters is primarily a function of carbonate,
bicarbonate, and hydroxide content, it is taken as an indication of the concentration of these
constituents. The measured values also may include contributions from borates, phosphates,
silicates, or other bases if these are present. Alkalinity in excess of alkaline earth metal
concentrations is significant in determining the suitability of water for irrigation. Alkalinity
measurements are used in the interpretation and control of water and wastewater treatment
processes. Raw domestic wastewater has an alkalinity less than, or only slightly greater than,
that of the water supply. Properly operating anaerobic digesters typically have supernatant
alkalinities in the range of 2000 to 4000 mg calcium carbonate (CaCO3)/L.
 Highly alkaline water is usually unpalatable.

 Large amount of alkalinity imparts bitter taste in water.
 The principal objection of alkaline water is the reactions that can occur between
alkalinity and certain cations in water. The resultant precipitate can foul pipes and
other appurtenances of water distribution systems.
 To neutralize acids produced during flocculation, the sample should be alkaline as
otherwise further floc formation ceases.
 To find out the quantity of lime and soda ash required for the removal of hardness,
alkalinity data is required.
 Wastewater containing excess caustic alkalinity is not to be discharged into natural
water bodies or sewers.
 Excess alkalinity in the water is harmful for irrigation, which leads to soil damage and
reduce crop yields.

 Alkalinity value is necessary in the calculation of carbonate scaling tendencies of
saline water.
 In wastewater treatment, alkalinity is an important parameter determining the
amenability of wastes to the treatment process and control of processes such as
anaerobic digestion, where bicarbonate alkalinity, total alkalinity and any fraction
contributed by volatile acid salts becomes considerations.
AIM
To determine the amount of alkalinity present in the given water sample.
PRINCIPLE
The alkalinity of water is a measure of its capacity to neutralize acids. It is primarily due to
salts of weak acids, although weak or strong bases may also contribute. Alkalinity is usually
imparted by bicarbonate, carbonate and hydroxide. It is measured volumetrically by titration
with 0.02 N sulphuric acid and is reported in terms of CaCO3 equivalent. For samples whose
initial pH is above 8.3, the titration is conducted in two steps. In the first step, the titration is
conducted until the pH is lowered to 8.3, the point at which phenolphthalein indicator turns
from pink to colourless. Decolourization of phenolphthalein indicator will show complete
neutralization of hydroxyl ions (Eq. 1) and half of carbonate ions (Eq. 2). The second phase
of titration is conducted until the pH is lowered to 4.5, corresponds to bromocresol green end
point, which corresponds to the equivalence points for the conversion of bicarbonate ion to
carbonic acid (Eq. 3).
(1)
(2)
(3)
APPARATUS
o Burette
o Erlenmeyer flask
o Pipettes

REAGENTS
 Carbon dioxide free distilled water.

 Phenolphthalein indicator.
 Mixed indicator.
 0.1 N sodium thiosulphate solution
 0.02 N sulphuric acid.
Mixed Indicator: Dissolve 100 mg bromcresol green sodium salt and 20 mg methyl red
sodium salt in 100 mL distilled water; or 95% ethyl alcohol or isopropyl alcohol.
Phenolphthalein Indicator: Add 1 g of phenolphthalein in 200 mL of distilled water or ethyl

alcohol. Then add 0.02 N NaOH in drop wise till pink colour disappears.
Standard Sulphuric Acid (0.02 N): Makeup 0.53 mL of 98% pure H2SO4 to one litre in a
volumetric flask and standardize it against 0.1 N sodium carbonate using methyl orange as
indicator.
PROCEDURE
1. Pipette 20 mL of sample into a clean Erlenmeyer flask (V).

2. Add one drop of sodium thiosulphate solution, if residual chlorine is present.
3. Add two drops of phenolphthalein indicator; if the pH is above 8.3, colour of solution
becomes pink. Note down the initial burette reading (V1).
4. Titrate against standard sulphuric acid in the burette, till the colour just disappears.
Note down the burette reading (V2). The volume of sulphuric acid consumed for the
phenolphthalein alkalinity (A) = V2-V1.
5. Then add two drops of mixed indicator, the colour turns yellow.
6. Again titrate against acid, until the colour turns to pink. Note down the burette reading
(V3). The volume of sulphuric acid consumed for the total alkalinity (B) = V3-V1.
CALCULATION

The results obtained from the phenolphthalein and total alkalinity determinations offer a
means for stoichiometric classification of the three principal forms of alkalinity present in
many waters. The classification ascribes the entire alkalinity to bicarbonate, carbonate, and
hydroxide, and assumes the absence of other (weak) inorganic or organic acids, such as
silicic, phosphoric, and boric acids.
 Carbonate alkalinity is present when phenolphthalein alkalinity is not zero but is less
than total alkalinity.
 Hydroxide alkalinity is present if phenolphthalein alkalinity is more than half the total
alkalinity.
 Bicarbonate alkalinity is present if phenolphthalein alkalinity is less than half the total
alkalinity.
The types of alkalinities present in the samples are calculated using the equations given in the
following table.
Alkalinity due to (mg/L as CaCO3)
Result of Titration
OH- CO32- HCO3-
P=0 0 0 T
P < 0.5 T 0 2P T-2P
P = 0.5 T 0 2P 0
P > 0.5 T 2P-T 2(T-P) 0
P=T T 0 0
OBSERVATIONS
Titration 1 (Phenolphthalein Alkalinity)

Burette solution: Sulphuric Acid
Indicator: Phenolphthalein
End Point: Disappearance of Pink Colour
Volume of Sample Burette Reading (mL) Volume of H2SO4
No.
(mL) Initial Final A (mL)

Titration 2 (Total Alkalinity)
Burette solution: Sulphuric Acid
Indicator: Mixed Indicator
End Point: Appearance of Pink Colour
Volume of Sample Burette Reading (mL) Volume of H2SO4
No.
(mL) Initial Final B (mL)
RESULT
Total alkalinity present in the given sample = ………………… mg/L as CaCO3

Alkalinity due to OH- = ………………… mg/L as CaCO3
Alkalinity due to CO32- = ………………… mg/L as CaCO3
Alkalinity due to HCO32- = ………………… mg/L as CaCO3
QUESTIONS
1. Which is the major form of alkalinity? How is it formed?

2. The water where algae are flourishing is alkaline. Why? Will there be diurnal
variation in the pH of such waters?
3. Why does the pH change on aerating the water?
4. For efficient coagulation the water must be alkaline. Why?
5. Why do we use CO2 free distilled water for analysis?
6. Explain the alkalinity titration curve for a hydroxide carbonate mixture.
7. With the help of a neat sketch, explain the relationship between carbon dioxide and
the three forms of alkalinity at various pH levels.
8. Explain carbonic acid system with the help of neat sketch.
9. Why alkalinity is expressed in calcium carbonate scale?
10. Calculate the alkalinity as CaCO3 of water that contains 85 mg/L of HCO3-, 120 mg/L
of CO32- and 2 mg/L of OH-.

DETERMINATION OF TOTAL HARDNESS
INTRODUCTION
Originally, the hardness of water was understood to be a measure of the capacity of water for
precipitating soap. Soap is precipitated chiefly by the calcium and magnesium ions
commonly present in water, but may also be precipitated by ions of other polyvalent metals,
such as aluminium, iron, manganese, strontium and zinc, and by hydrogen ions. Because, all
but the first two are usually present in insignificant concentrations in natural waters, hardness
is defined as a characteristic of water, which represents the total concentration of just the
calcium and the magnesium ions expressed as calcium carbonate. However, if present in
significant amounts, other hardness producing metallic ions should be included.
When the hardness is numerically greater than the sum of the carbonate alkalinity and the
bicarbonate alkalinity, the amount of hardness, which is equivalent to the total alkalinity, is
called carbonate hardness; the amount of hardness in excess of this is called non-carbonate
hardness. When the hardness is numerically equal to or less than the sum of carbonate and
bicarbonate alkalinity all of the hardness is carbonate hardness and there is no non- carbonate
hardness. The hardness may range from zero to hundreds of milligrams per litre in terms of
calcium carbonate, depending on the source and treatment to which the water has been
subjected.
 Absolutely soft water is tasteless. On the other hand, hardness up to 600 mg/L can be
relished if got acclimatized to.
 Even though the scales formed as inner coating of the pipelines prevent corrosion, it
reduces the carrying capacity. The scale formation causes enormous loss of fuel in
boilers.
 Absolutely soft water is corrosive and dissolves the metals.
 More cases of cardio vascular diseases are reported in soft water area.
 Hard water is useful for the growth of children due to the presence of calcium.
 Hard water cause excessive consumption of soap used for the cleaning purpose.
 Magnesium hardness, particularly associated with sulphate ion has laxative effects on
persons unaccustomed to it.

 It affects the working of dyeing process.
 It also precipitates protein of meat and make it tasteless.
AIM
To determine the amount of hardness present in the given water sample.
PRINCIPLE
Ethylenediamine tetra-acetic acid and its sodium salts (EDTA) form a chelated soluble
complex with calcium and magnesium ions and other divalent cations causing hardness (Eq.
1). The successful use of EDTA for determining hardness depends upon an indicator present
to show when EDTA present in excess, or when all the ions causing hardness have been
completed. If a small amount of a dye such as Eriochrome black T (have a blue colour) is
added to an aqueous solution containing calcium and magnesium ions at a pH of 10 ± 0.1, the
solution will become wine red (Eq. 2). If EDTA is then added as a titrant, all free hardness
ions form complex with EDTA. Finally, the EDTA disrupts the wine red complex (M. EBT)
because it is capable of forming more suitable complex with the hardness ions. This action
frees the EBT indicator, and wine red colour changes to a distinct blue colour, heralding the
end point of titration.
(1)
(2)
APPARATUS
o Burette
o Erlenmeyer flask
o Pipettes
o Beakers
REAGENTS
 Ammonium buffer solution

 Standard EDTA titrant (0.02 N)
 Eriochrome black T indicator

Ammonia Buffer Solution: Dissolve 16.9 g ammonium chloride (NH4Cl) in 143 mL

concentrated ammonium hydroxide (NH4OH). Add 1.25 g magnesium salt of EDTA
(available commercially) and dilute to 250 mL with distilled water.
If the magnesium salt of EDTA is unavailable, dissolve 1.179 g disodium salt of EDTA
(analytical reagent grade) and 780 mg magnesium sulfate (MgSO4⋅7H2O) or 644 mg
magnesium chloride (MgCl2⋅6H2O) in 50 mL distilled water. Add this solution to 16.9 g
NH4Cl and 143 mL concentrated NH4OH with mixing and dilute to 250 mL with distilled
water.
Eriochrome Black T Indicator: Dissolve 0.5 g EBT in 100 g 2,2′,2′′-nitrilotriethanol (also

called triethanolamine) or 2-methoxymethanol (also called ethylene glycol monomethyl
ether). Add 2 drops per 50 mL solution to be titrated.
Standard EDTA Solution (0.02 N): Weigh 3.723 g analytical reagent-grade di-sodium
EDTA, also called (ethylenedinitrilo)tetraacetic acid disodium salt (EDTA), dissolve in
distilled water, and dilute to 1000 mL. Standardize against standard calcium solution.
PROCEDURE
1. Take 20 mL of sample in an Erlenmeyer flask.

2. Add 2 mL of ammonia buffer solution.
3. Add two drops of indicator solution. The solution turns wine red in colour.
4. Add the standard EDTA titrant slowly with continuous stirring until the last reddish
tinge disappears from the solution. The colour of the solution at the end point is blue
under normal conditions.
5. Note down the volume of EDTA added (V).
CALCULATION

OBSERVATIONS
Burette solution: EDTA

Indicator: EBT
End Point: Wine Red to Blue
Volume of Sample Burette Reading (mL) Volume of EDTA
No.
(mL) Initial Final (mL)
RESULT
The amount of total hardness present in the given sample = ……………… mg/L as CaCO3
QUESTIONS
1. What is degree of hardness? How will you classify water in terms of degree of
hardness?
2. Can you determine temporary hardness and permanent hardness separately? If yes,
how?
3. What are the principal cations causing hardness in water and the major anions
associated with them?
4. How is hardness classified? Explain their removal methods.
5. Why is softening of water necessary? What are the advantages of soft water?
6. What are the differences between scum and scale?
7. What is a scale? What is the main cause for scale formation? Temporary or permanent
hardness? Support your answer with suitable chemical equations.
8. Explain the relationship between hardness and alkalinity.
9. What is pseudo-hardness?
10. Report the level of groundwater hardness in India (At least for 10 Indian cities).

DETERMINATION OF CALCIUM HARDNESS
INTRODUCTION
Hard water is generally considered as those water that require considerable amount of soap to
produce a foam or lather and that also produce scale in hot water pipes, heaters, boilers and
other units in which temperature of water is increased materially. Calcium is one of the
principal cations involved in water hardness and usually found in higher concentration in
natural water. Lime stone, gypsum etc. are responsible for the higher calcium deposits in
water. The calcium content may range from zero to several hundreds ppm.
AIM
To determine the amount of calcium hardness present in the given water sample.
PRINCIPLE
Ethylenediamine tetra-acetic acid and its sodium salts (EDTA) form a chelated soluble
complex with calcium and magnesium ions and other divalent cations causing hardness. In
order to suppress the complex reaction between EDTA and magnesium ions, the solution is
titrated at elevated pH. Because, magnesium precipitates as magnesium hydroxide at highly
alkaline condition, the remaining calcium in water forms complex with EDTA. An indicator,
ammonium purpurate which combines only with calcium is used. The indicator imparts a
pink colour to the solution after making complex with calcium ions. If EDTA is then added
as a titrant, all free calcium ions form complex with EDTA. Finally, the EDTA disrupts the
pink complex (Ca. ammonium purpurate) because it is capable of forming more suitable
complex with the calcium ions. This action frees the ammonium purpurate indicator, and pink
colour changes to a distinct purple colour, heralding the end point of titration.
APPARATUS
o Burette
o Erlenmeyer flask
o Pipettes
o Beakers

REAGENTS
 Sodium hydroxide solution (1 N)

 Standard EDTA titrant (0.02 N)
 Ammonium purpurate indicator
Ammonium Purpurate: Mix 0.5 g of ammonium purpurate and 100 g NaCl. Use it as dry
powder.
Sodium Hydroxide (1 N): Dissolve 40 g NaOH in 1 litre of distilled water.
Standard EDTA Solution (0.02 N): Weigh 3.723 g analytical reagent-grade di-sodium
EDTA, also called (ethylenedinitrilo) tetra acetic acid disodium salt (EDTA), dissolve in
distilled water, and dilute to 1000 mL. Standardize against standard calcium solution.
PROCEDURE
1. Take 20 mL of sample in an Erlenmeyer flask.

2. Add 1 mL of 1 N NaOH solution.
3. Add a pinch of ammonium purpurate powder to the solution. The solution turns pink
in colour.
4. Add the standard EDTA titrant slowly with continuous stirring until the last pinkish
tinge disappears from the solution. The colour of the solution at the end point is
purple under normal conditions.
5. Note down the volume of EDTA added (V).
CALCULATION

OBSERVATIONS
Burette solution: EDTA

Indicator: Ammonium Purpurate
End Point: Pink to Purple
Volume of Sample Burette Reading (mL) Volume of EDTA
No.
(mL) Initial Final (mL)
RESULT
Total amount of calcium hardness present in the given sample = ……………mg/L as CaCO3
= ………………. mg/L as Ca

DETERMINATION OF DISSOLVED OXYGEN
INTRODUCTION
All living organisms depend upon oxygen in one form or another to maintain the metabolic
processes that produce energy for the growth and reproduction. Environmental engineers and
scientists are, of course, interested in atmospheric conditions in relations to humans, but, in
addition, they are vitally concerned with the “atmospheric conditions” that exists in liquids,
water being the liquid in greatest abundance and importance.
All the gases of atmosphere are soluble in water to some degree. Both nitrogen and oxygen
are classified as poorly soluble, and since they do not react with water chemically, their
solubility is directly proportional to their partial pressure. The solubility of atmospheric
oxygen in fresh water ranges from 14. 6 mg/L at 0oC to 7 mg/L at 35oC under 1 atm of
pressure.
The low solubility of oxygen is the major factor that limits the purification capacity of natural
water and necessitates treatment of wastes to remove polluting matter before discharge into
receiving streams. In polluted water the saturation value of Oxygen is also less than that of
clean water. The ratio of the value in polluted water to that of clean water is referred to as β
value. The rate of absorption of oxygen in polluted water is normally less than in clean water
and the ratio is referred as α value. They may range as low as 0.8 for β and 0.4 for α in some
wastewater. Both α and β are important design factors in selection of aeration equipment.
 The minimum DO level of 4 mg/L is desirable for the survival of aquatic life.
 Higher values of DO may cause corrosion of iron and steel.
 Drinking water should be rich in DO for good taste.
 Higher temperature, biological impurities, ammonia, nitrates, ferrous ion, hydrogen
sulphide and organic matter reduce DO values.
 It is necessary to know DO levels to assess quality of raw water and to keep a check
on stream pollution.
 DO test is the basis of BOD test which is an important parameter to evaluate organic
pollution potential of waste.

 DO test is necessary for all aerobic biological wastewater treatment process to control
the rate of aeration.
AIM
To determine the amount of dissolved oxygen present in the given water sample.
PRINCIPLE
The Azide modified Winkler method is used to determine the DO concentration present in the
water sample. In Winkler method, in the absence of oxygen, a pure white precipitate of
Mn(OH)2 forms when MnSO4 and alkali iodide reagent are added to the sample (Eq. 1). If
oxygen is present in the sample, some of the Mn2+ is oxidized to Mn4+ (Eqns. 2and 3) and
precipitates as a brown hydrated oxide (Eq. 4). The oxidation of Mn2+ to MnO2, is sometimes
called fixation of oxygen. MnO(OH)2 appears as brown precipitate. Some sources claim that
Mn(OH)3 is the brown precipitate, but hydrated MnO2 also give brown precipitate.
(1)
(2)
(3)
(4)
Under the low pH conditions, the iodide ions added is oxidized to iodine (Eq. 5). Standard
thiosulphate is used, with a starch indicator; to titrate the iodine released (Eq. 6). This iodine
is equivalent to DO present in the sample.
(5)
(6)
The nitrate ion is one of the most frequent interferences encountered in the DO
determination. It doesn’t oxidize Mn2+, but does oxidize I- to I2 under acidic conditions (Eq.
7). It is particularly troublesome because its reduced form N2O2, is oxidized by oxygen,
which enters the sample during the titration procedure and is converted to NO2- again (Eq. 8),
establishing a cyclic reaction that can lead to erroneously high results, far in excess of
amounts that could be expected. Nitrate interference may be easily overcome by the use of

sodium azide. It is most convenient to incorporate the azide in the alkali iodide reagent. The
azide destroys nitrite in the presence of sulphuric acid and suppresses the interference caused
by nitrite (Eqns. 9 and 10).
(7)
(8)
(9)
(10)
APPARATUS
o Burette
o Erlenmeyer flask
o Pipettes
o Beakers
o 300 mL capacity BOD bottle
REAGENTS
 Manganous sulphate solution

 Alkali-iodide azide reagent
 Conc. sulphuric acid
 Starch indicator
 Standard sodium thiosulphate solution (0.025N)
Manganous Sulphate Solution: Dissolve 480 g MnSO4⋅4H2O, 400 g MnSO4⋅2H2O, or 364 g

MnSO4⋅H2O in distilled water, filter, and dilute to 1 L. The MnSO4 solution should not give a
colour with starch when added to an acidified potassium iodide (KI) solution.
Alkali-Iodide-Azide Reagent: Dissolve 500 g NaOH (or 700 g KOH) and 135 g NaI (or 150
g KI) in distilled water and dilute to 1 L. Add 10 g NaN3 dissolved in 40 mL distilled water.
Potassium and sodium salts may be used interchangeably. This reagent should not give a
colour with starch solution when diluted and acidified.

Starch Indicator: Dissolve 2 g laboratory-grade soluble starch and 0.2 g salicylic acid, as a
preservative, in 100 mL hot distilled water.
Standard Sodium Thiosulfate Titrant: Dissolve 6.205 g Na2S2O3⋅5H2O in distilled water.

Add 1.5 mL 6N NaOH or 0.4 g solid NaOH and dilute to 1000 mL. Standardize with bi-
iodate solution.
PROCEDURE
1. Add 2 mL of manganous sulphate solution and 2 mL of alkali-iodide azide reagent to

the 300 mL sample taken in the bottle, well below the surface of the liquid. (The
pipette should be dipped inside the sample while adding the above two reagents.)
2. Stopper with care to exclude air bubbles and mix by inverting the bottle at least 15
times.
3. When the precipitate settles, leaving a clear supernatant above the manganese
hydroxide floc, shake again.
4. After 2 minutes of settling, carefully remove the stopper, immediately add 2 mL
concentrated sulphuric acid by allowing the acid to run down the neck of the bottle.
5. Restopper and mix by gentle inversion until dissolution is complete.
6. Measure out 203 mL of the solution from the bottle to an Erlenmeyer flask. As 2 mL
each of manganese sulphate and azide reagent have been added, the proportionate
quantity of yellow solution corresponding to 200 mL of sample is
7. Titrate with 0.025 N sodium thiosulphate solution to a pale straw colour.

8. Add 1–2 mL starch solution and continue the titration to the first disappearance of the
blue colour and note down the volume of sodium thiosulphate solution added (V),
which gives directly the D.O. in mg/L.
CALCULATION
Titration of a sample of size equivalent to 200 mL of the original sample with 0.025 N
Na2S2O3 yield results in mL, which can be interpreted directly in terms of mg/L of DO.

OBSERVATIONS
Burette Solution: Sodium Thiosulphate (0.025 N)

Indicator: Starch
End Point: Blue to Colourless
Volume of Sample Burette Reading (mL) Volume of Sodium
No.
(mL) Initial Final Thiosulphate (mL)
RESULT
Total amount of dissolved oxygen present in the given sample = ……………mg/L
QUESTIONS
1. Vigorous shaking of samples is required after the addition of alkali iodide reagent and
MnSO4. Why?
2. In Winkler method, the blue colour of the starch indicator return even after the
addition of higher amount of sodium thiosulphate. What is the reason behind this?
How can we rectify this?
3. Most of the critical conditions related to dissolved oxygen deficiency occur during
summer months. Why?
4. The turbulence of water should be encouraged. Why?
5. Explain self purification of streams. Draw the oxygen sag curve.
6. List out the saturation DO values at various temperatures.
7. What is oxygen fixation? Give two reasons why fixation of DO should be performed
in the field if at all feasible.

8. Two samples were collected simultaneously at the same spot in a river for DO
analysis. One sample was “fixed” immediately after collection, and the other was
treated later in the laboratory. Indicate two possible factors that could cause lower
results to be obtained in the second sample.
9. Prepare a table showing five substances that interfere with the Winkler method,
explaining their interferences.
10. Explain the differences between reoxigenation and deoxigenation.
11. What are the factors and processes affecting the DO concentration in water?
12. Explain Henry’s law.

DETERMINATION OF BIOCHEMICAL OXYGEN DEMAND
INTRODUCTION
The Biochemical Oxygen Demand (BOD) of sewage or of polluted water is the amount of
oxygen required for the biological decomposition of dissolved organic matter to occur under
aerobic condition and at the standardized time and temperature. Usually, the time is taken as
5 days and the temperature 20°C as per the global standard.
The BOD test is among the most important method in sanitary analysis to determine the
polluting power, or strength of sewage, industrial wastes or polluted water. It serves as a
measure of the amount of clean diluting water required for the successful disposal of sewage
by dilution. The test has its widest application in measuring waste loading to treatment plants
and in evaluating the efficiency of such treatment systems.
 BOD is the principal test to give an idea of the biodegradability of any sample and
strength of the waste. Hence the amount of pollution can be easily measured by it. It is
the basic criteria for the control of the stream pollution.
 Efficiency of any treatment plant can be judged by considering influent BOD and the
effluent BOD.
 Ordinary domestic sewage may have a BOD of 200 mg/L. Any effluent to be
discharged into natural water bodies should have BOD less than 30 mg/L.
 Drinking water usually has a BOD less than 1 mg/L. But, when BOD value reaches 5
mg/L, the water is doubtful in purity.
 The determination of BOD is used in studies to measure the self purification capacity
of streams and serves regulatory authorities as a means of checking on the quality of
effluents discharged to such water.
 It is a factor in the choice of treatment method and is used to determine the size of
certain units, particularly trickling filter and activated sludge units.
 It is useful to estimate the population equivalent of any industrial pollutant which is
useful to collect cess from the industrialist for purification of industrial wastes in
municipal sewage treatment plant.

AIM
To determine the BOD value of the given water sample.
PRINCIPLE
The test consists in taking the given sample in suitable concentrations in dilute water in BOD
bottles. Two bottles are taken for each concentration and three concentrations are used for
each sample. One set of bottles is incubated in a BOD incubator for 5 days at 20°C; the
dissolved oxygen (initial) content (D1) in the other set of bottles will be determined
immediately. At the end of 5 days, the dissolved oxygen content (D2) in the incubated set of
bottles is determined.
where, P = decimal fraction of sample used.
D1 = dissolved oxygen of diluted sample (mg/L), immediately after preparation.
D2 = dissolved oxygen of diluted sample (mg/L), at the end of 5 days incubation.
Among the three values of BOD obtained for a sample select that dilution showing the
residual dissolved oxygen of at least 1 mg/L and a depletion of at least 2 mg/L. If two or
more dilutions are showing the same condition then select the BOD value obtained by that
dilution in which the maximum dissolved oxygen depletion is obtained.
APPARATUS
o BOD bottles 300mL capacity

o BOD incubator
o Burette
o Pipette
o Air compressor
o Measuring cylinder etc.
REAGENTS
 Distilled water
 Phosphate buffer solution
 Magnesium sulphate solution
 Calcium chloride solution
 Ferric chloride solution
 Acid and alkali solution
 Sodium sulphite solution
 Reagents required for the determination of DO.
Phosphate Buffer Solution: Dissolve 8.5 g KH2PO4, 21.75 g K2HPO4, 33.4 g

Na2HPO4⋅7H2O, and 1.7 g NH4Cl in about 500 mL distilled water and dilute to 1 L. The pH
should be 7.2 without further adjustment. Alternatively, dissolve 42.5 g KH2PO4 or 54.3 g
2HPO4 in about 700 mL distilled water. Adjust pH to 7.2 with 30% NaOH and dilute to 1 L.
Magnesium Sulfate Solution: Dissolve 22.5 g MgSO4⋅7H2O in distilled water and dilute to
1 L.
Calcium Chloride Solution: Dissolve 27.5 g CaCl2 in distilled water and dilute to 1 L.
Ferric Chloride Solution: Dissolve 0.25 g FeCl3⋅6H2O in distilled water and dilute to 1 L.
Dilution Water: The BOD concentration in most wastewaters exceeds the concentration of
dissolved oxygen (DO) available in an air-saturated sample. Therefore, it is necessary to
dilute the sample before incubation to bring the oxygen demand and supply into appropriate
balance. Because bacterial growth requires nutrients such as nitrogen, phosphorus, and trace
metals, these are added to the dilution water, which is buffered to ensure that the pH of the
incubated sample remains in a range suitable for bacterial growth. Complete stabilization of a
sample may require a period of incubation too long for practical purposes; therefore, 5 day
has been accepted as the standard incubation period. If the dilution water is of poor quality,
the BOD of the dilution water will appear as sample BOD. This effect will be amplified by
the dilution factor. High quality organic free water must be used for dilution water.
Aerate required volume of water with a supply of clean compressed air. Add 1 mL each of
calcium chloride, Magnesium sulphate, ferric chloride and phosphate buffer solution to 1 L of
aerated distilled water and mix thoroughly. This is the standard dilution water.

PROCEDURE
1. Place the desired volume of distilled water in a 5 litre flask (usually about 3 L of
distilled water will be needed for each sample).
2. Add 1mL each of phosphate buffer, magnesium sulphate solution, calcium chloride
solution and ferric chloride solution for every litre of distilled water.
3. Seed the dilution water with 1–2 mL of settled domestic sewage.
4. Saturate the dilution water in the flask by aerating with a supply of clean compressed
air for at least 30 minutes.
5. Highly alkaline or acidic samples should be neutralized to pH 7.
6. Destroy the chlorine residual in the sample by keeping the sample exposed to air for 1
to 2 hours or by adding a few mL of sodium sulphite solution.
7. Take the sample in the required concentrations. The following concentrations are
suggested:
Strong industrial waste: 0.1, 0.5 and 1 per cent
Raw and settled sewage: 1.0, 2.5 and 5 per cent
Oxidised effluents: 5, 12.5 and 25 per cent
Polluted river water: 25, 50 and 100 per cent
8. Add the required quantity of sample (calculate for 650 mL dilution water the required
quantity of sample for a particular concentration) into a 1000 mL measuring cylinder.
Add the dilution water up to the 650 mL mark.
9. Mix the contents in the measuring cylinder.
10. Add this solution into two BOD bottles, one for incubation and the other for
determination of initial dissolved oxygen in the mixture.
11. Prepare in the same manner for other concentrations and for all the other samples.
12. Lastly fill the dilution water alone into two BOD bottles. Keep one for incubation and
the other for determination of initial dissolved oxygen.
13. Place the set of bottles to be incubated in a BOD incubator for 5 days at 20°C. Care
should be taken to maintain the water seal over the bottles throughout the period of
incubation.
14. Determine the initial dissolved oxygen contents in the other set of bottles and note
down the results.
15. Determine the dissolved oxygen content in the incubated bottles at the end of 5 days
and note down the results.
16. Calculate the B.O.D. of the given sample.
CALCULATION
where,
D1= Initial DO of sample
D2= Final DO of sample (after incubation period)
BC = blank correction = C1-C2
C1 = Initial DO of dilution water
C2 = DO of dilution water at the end of 5 days.
OBSERVATIONS

Indicator: Starch
Volume of Sample Burette Reading (mL) Volume of Sodium
No.
(mL) Initial Final Thiosulphate (mL)
Initial Day
Final Day

RESULT
BOD of the given sample = ……………mg/L
QUESTIONS
1. What are the three methods that can be used to control nitrification in the 5 days BOD
test at 20°C?
2. What are the factors affecting the rate of biochemical oxidation in the BOD test?
3. List five requirements, which must be complied with, in order to obtain reliable BOD
data.
4. Explain how a sample of river water having a temperature below 20oC should be pre-
treated in preparation for BOD analysis.
5. What method can be used to control nitrification in 5 day BOD test?
6. With the help of a neat sketch, explain CBOD and NBOD in detail.
7. Why 5 days are selected for the standard BOD analysis?
8. A student began experiment for determination of 5-day, 20C BOD on Monday. Since
the 5th day fell on Saturday, the final DO readings were taken on Monday. On
calculation the 7-day BOD was found to be 150 mg/L. What would be the 5-day BOD
in mg/L. Assume BOD rate constant (k) at 20C as 0.23/day(base e).
9. Explain Thomas slope method and Fujimotto method for finding the ultimate BOD.
10. Standard BOD test has been carried out for a water sample having initial temperature
of 32oC and the following readings are observed. Find the BOD of the sample after 7
days at 32oC. Initial DO of the sample is 8.7 mg/L.
Sample volume (mL) 2 5 7 12
DO after 5 days (mg/L) 6.9 5.2 4.4 0.9

DETERMINATION OF CHEMICAL OXYGEN DEMAND
INTRODUCTION
The Chemical Oxygen Demand (COD) test is widely used as means of measuring the organic
strength of domestic and industrial wastes and is defined as the amount of a specified oxidant
that reacts with the sample under controlled conditions. The quantity of oxidant consumed is
expressed in terms of its oxygen equivalence. Both organic and inorganic components of a
sample are subject to oxidation, but in most cases the organic component predominates and is
of the greater interest. If it is desired to measure either organic or inorganic COD alone,
additional steps not described here must be taken to distinguish one from the other. COD is a
defined test; the extent of sample oxidation can be affected by digestion time, reagent
strength, and sample COD concentration.
 The COD test is used extensively in the analysis of industrial wastewater.

 The test is 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 the strength of wastes, which contain toxins and biologically
resistant organic substances.
 The ratio of BOD to COD is useful to assess the amenability of waste for biological
treatment. Ratio of BOD to COD greater than or equal to 0.8 indicates wastewater
highly amenable to the biological treatment.
 COD value is always greater than BOD value. For domestic and some industrial
wastewater, COD value is about 2.5 times BOD value.
AIM
To determine the COD value of the given water sample.
PRINCIPLE
The organic matter present in the sample gets oxidized completely by K2Cr2O7 in the
presence of H2SO4 to produce CO2 and H2O. The excess K2Cr2O7 remaining after the
reaction is titrated with ferrous ammonium sulphate, using ferroin as indicator. The

dichromate consumed by the sample is equivalent to the amount of oxygen required to
oxidize the organic matter.
APPARATUS
o COD digester
o COD vials with stand
o Burette
o Pipette
o Conical flask etc.
REAGENTS
 Standard potassium dichromate

 Sulphuric acid
 Standard ferrous ammonium sulphate
 Ferroin indicator
 Mercury sulphate
 Silver sulphate
 Organic free distilled water
Standard Potassium Dichromate Digestion Solution (0.01667 M): Add to about 500 mL
distilled water 4.903 g K2Cr2O7, primary standard grade, previously dried at 150°C for 2 h,
167 mL concentrated H2SO4, and 33.3 g HgSO4. Dissolve, cool to room temperature, and
dilute to 1000 mL.
Sulfuric Acid Reagent: Add Ag2SO4, reagent or technical grade, crystals or powder, to
concentrated H2SO4 at the rate of 5.5 g Ag2SO4/kg H2SO4. Let stand 1 to 2 days to dissolve.
Mix.
Ferroin Indicator Solution: Dissolve 1.485 g 1,10-phenanthroline monohydrate and 695 mg

FeSO4⋅7H2O in distilled water and dilute to 100 mL.
Standard Ferrous Ammonium Sulfate Titrant (FAS), Approximately 0.10M: Dissolve

39.2 g Fe(NH4)2(SO4)2⋅6H2O in distilled water. Add 20 mL concentrated H2SO4, cool, and
dilute to 1000 mL.

PROCEDURE
1. To a 2.5 mL sample taken in a COD vial, add 1.5 mL of K2Cr2O7 reagent and 3.5 mL
of H2SO4 reagent carefully.
2. Employ a hot blank (Distilled water is taken instead of sample).
3. Switch on the COD digester and fix the temperature at 150oC and set the time at 2h.
4. Place the vials in the digester and heat for 2 h at 150oC.
5. Cool the mixture to room temperature and titrate the excess dichromate with FAS
using ferroin indicator.
6. The end point is the sharp colour change from blue-green to reddish brown, although
the blue-green colour may reappear within minutes.
7. Repeat the same procedure for blank solution.
CALCULATION
where, A = Volume of FAS used for blank ; B = Volume of FAS used for sample; N =
Normality of FAS.
OBSERVATIONS
Burette Solution: Ferrous Ammonium Sulphate

Indicator: Ferroin
End Point: Blue-green to Reddish Brown
Titration 1 (Distilled water)
Volume of Sample Burette Reading (mL) Volume of FAS
No.
(mL) Initial Final ‘A’ (mL)

Titration 2 (Sample)
Volume of Sample Burette Reading (mL) Volume of FAS

No.
(mL) Initial Final ‘B’ (mL)
RESULT
COD of the given sample = ……………mg/L
QUESTIONS
1. What general groups of organic compounds are not oxidized in COD analysis?
2. What is the most prevalent inorganic species that interferes with the COD analysis
and how is this interference dealt with?
3. Why do the COD and BOD analyses usually give different results for the same
waste?
4. Indicate whether COD results would probably be higher, lower or the same as the
true value under the following conditions and briefly explain why: (a) HgSO4 was
not added (b) AgSO4 was not added (c) the FAS was assumed to have the same
normality as it did two weeks prior to the current analysis.
5. What would be inferred from the following analytical results concerning the relative
ease of biodegradability of each waste?
Waste BOD5,20 (mg/L) COD (mg/L)
A 240 300
B 100 500
C 120 240

DETERMINATION OF IRON
INTRODUCTION
Iron (Fe) is the first element in Group VIII of the periodic table; it has an atomic number of
26, an atomic weight of 55.85, and common valencies of 2 and 3 (and occasionally valencies
of 1, 4, and 6). The average abundance of Fe in the earth’s crust is 6.22%; in soils Fe ranges
from 0.5 to 4.3%; in streams it averages about 0.7 mg/L; and in groundwater it is 0.1 to 10
mg/L. Iron occurs in the minerals hematite, magnetite, taconite, and pyrite. It is widely used
in steel and in other alloys.
The solubility of ferrous ion (Fe2+) is controlled by the carbonate concentration. Because
groundwater is often anoxic, any soluble iron in groundwater is usually in the ferrous state.
On exposure to air or addition of oxidants, ferrous iron is oxidized to the ferric state (Fe3+)
and may hydrolyze to form red, insoluble hydrated ferric oxide. In the absence of complex-
forming ions, ferric iron is not significantly soluble unless the pH is very low.
Iron and manganese are two similar elements that can be a nuisance in a drinking water
supply. Iron is more common than manganese, but they often occur together. They are not
hazardous to health.
Iron and manganese are common elements in the earth’s crust. As water percolates through
soil and rock it can dissolve these minerals and carry them into groundwater. Also, iron pipes
can corrode and leach iron into a household water supply.
 Although iron has got little concern as a health hazard, but still is considered as a
nuisance in excessive quantities. Long time consumption of drinking water with a
high concentration of iron can lead to liver diseases (hemosiderosis).
 Iron causes reddish-brown stains on laundry, porcelain, dishes, utensils, glass-ware,
sinks, fixtures and concrete. Detergents do not remove these stains. Chlorine bleach
and alkaline builders (such as sodium and carbonate) may even intensify the stains.
 Iron also forms red spot on clothes.
 Iron rich water exposed to the air, become turbid and highly unacceptable from the
aesthetic point of view, due to the oxidation of soluble ferrous ion to insoluble ferric
hydroxides or oxides, which settles out as a rust coloured salt. Such water often tastes
unpalatable even at low concentration.
 Iron promotes the growth of iron bacteria which derive their energy from the
oxidation of ferrous to ferric. This gives rusty appearance to the water. Colonies of
these bacteria also form a slime which causes problems in water closets, pipes, pumps
and distribution systems.
 High concentration iron in water is not suitable for processing of food, beverages, ice,
dyeing, bleaching etc. Water with higher iron concentration when used in the
preparation of tea and coffee, interacts with tannins giving a black inky appearance
with a metallic taste. Coffee may become unpalatable at concentration of iron more
than 1 mg/L.
AIM
To determine the amount of iron present in the given water sample.
PRINCIPLE
Iron is brought into solution, reduced to the ferrous state by boiling with acid and
hydroxylamine, and treated with 1,10-phenanthroline at pH 3.2 to 3.3. Three molecules of
phenanthroline chelate each atom of ferrous iron to form an orange-red complex. The
coloured solution obeys Beer’s law; its intensity is independent of pH from 3 to 9. A pH
between 2.9 and 3.5 insures rapid colour development in the presence of an excess of
phenanthroline. Colour standards are stable for at least 6 months.
APPARATUS
o Colourimetric equipment; one of the following is required:

a) Spectrophotometer, for use at 510 nm, providing a light path of 1 cm or longer.
b) Nessler tubes, matched, 100 mL, tall form.
o Glassware like conical flasks, pipettes and glass beads.
REAGENTS
 Hydrochloric acid
 Hydroxylamine solution
 Ammonium acetate buffer solution
 Sodium acetate solution
 Phenanthroline solution
 Stock iron solution
 Standard iron solution (1 mL = 10 µg Fe)
Hydrochloric Acid: Concentrated HCl containing less than 0.5 ppm iron.
Hydroxylamine Solution: Dissolve 10 g NH2OH⋅HCl in 100 mL water.
Ammonium Acetate Buffer Solution: Dissolve 250 g NH4C2H3O2 in 150 mL water. Add
700 mL concentrated (glacial) acetic acid. Because even a good grade of NH4C2H3O2
contains a significant amount of iron, prepare new reference standards with each buffer
preparation.
Sodium Acetate Solution: Dissolve 200 g NaC2H3O2⋅3H2O in 800 mL water.
Phenanthroline Solution: Dissolve 100 mg 1,10-phenanthroline monohydrate,

C12H8N2⋅H2O, in 100 mL water by stirring and heating to 80°C. Do not boil. Discard the
solution if it darkens. Heating is unnecessary if 2 drops concentrated HCl are added to the
water.
Potassium Permanganate (0.1 M): Dissolve 0.316 KMnO4 in reagent water and dilute to
100 mL.
Stock Iron Solution: Slowly add 20 mL concentrated H2SO4 to 50 mL water and dissolve
1.404 g Fe(NH4)2(SO4)2⋅6H2O. Add 0.1M potassium permanganate (KMnO4) dropwise until
a faint pink colour persists. Dilute to 1000 mL with water and mix; 1 mL = 200 µg Fe.
Standard Iron Solutions: Pipet 50 mL stock solution into a 1000 mL volumetric flask and
dilute to mark with water; 1 mL = 10 µg Fe.
PROCEDURE
Preparation of standards and blank:
1. Pipette 10, 20, 30 and 50 mL. Standard iron solution into 100 mL conical flasks.

2. Add 2 mL concentrated HCl and 1 mL hydroxylamine solution to each flask.
3. Dilute each to about 75 mL with distilled water.
4. Add a few glass beads and heat to boiling. To insure dissolution of all the iron,
continue boiling until volume is reduced to 15 to 20 mL.
5. Cool to room temperature and transfer to 100-mL Nessler tube.
6. Add 10 mL ammonium acetate buffer solution and 4 mL phenanthroline solution, and
dilute to 100 mL mark with distilled water.
7. Mix thoroughly and allow a minimum of 10 min for maximum colour development.
8. Take 50 mL distilled water in another conical flask and repeat steps 2 to 7 described
above.
Calibration:
9. Measure the absorbance of each solution in a spectrophotometer at 508 nm against the

reference blank prepared by treating distilled water as described in step 8. Prepare a
calibration graph taking meter reading on y-axis and concentration of iron on x-axis.
10. For visual comparison, keep Nessler tubes in a stand.
Sample analysis:
11. Mix the sample thoroughly and measure 50 mL into a conical flask and repeat steps 2
to 7 described above.
12. Measure the absorbance of the solution in a 1cm cell in a spectrophotometer at 508
nm.
13. Read off the concentration of iron (µg Fe) from the calibration graph for the
corresponding meter reading.
14. For visual comparison, match the colour of the sample with that of the standard
prepared in steps 1 to 7 above.
15. The matching colour standard will give the concentration of iron in the sample (µg
Fe).
CALCULATION

OBSERVATIONS
Spectrophotometer Analysis
Volume of Iron Content from Concentration of Iron

No. Absorbance
Sample (mL) Graph (µg) (mg/L)
Visual Comparison
Volume of Concentration of Iron in Matching Concentration of Iron
No.
Sample (mL) Colour Standard (µg) (mg/L)
RESULT
The amount of iron present in the given sample is = .................................................. mg/L
QUESTIONS
1. How are the iron and manganese removed from water?

2. Discuss the different oxidation states of iron occurring in natural water supplies, and
discuss the condition under which each form is stable.
3. Discuss briefly how iron and manganese get into underground water supplies.
4. In what form must iron exist in order to be measured by the phenanthroline method,
and how is iron converted to this form?

DETERMINATION OF MANGANESE
INTRODUCTION
Manganese (Mn) is the first element in Group VIIB in the periodic table; it has an atomic
number of 25, an atomic weight of 54.94, and common valencies of 2, 4, and 7 (and more
rarely, valencies of 1, 3, 5, and 6). The average abundance of Mn in the earth’s crust is 1060
ppm; in soils it is 61 to 1010 ppm; in streams it is 7 µg/L, and in groundwater it is <0.1 mg/L.
Manganese is associated with iron minerals, and occurs in nodules in ocean, fresh waters, and
soils. The common ores are pyrolusite (MnO2) and psilomelane. Manganese is used in steel
alloys, batteries, and food additives.
The common aqueous species are the reduced Mn2+ and the oxidized Mn4+. The aqueous
chemistry of manganese is similar to that of iron. Since groundwater is often anoxic, any
soluble manganese in groundwater is usually in the reduced state (Mn2+). Upon exposure to
air or other oxidants, groundwater containing manganese usually will precipitate black MnO2.
Elevated manganese levels therefore can cause stains in plumbing/laundry, and cooking
utensils. It is considered an essential trace element for plants and animals.
 Iron and manganese can give water an unpleasant taste, odour and colour.
 Manganese causes brownish-black stains on laundry, porcelain, dishes, utensils, glass-
ware, sinks, fixtures and concrete. Detergents do not remove these stains. Chlorine
bleach and alkaline builders (such as sodium and carbonate) may even intensify the
stains.
 Iron and manganese deposits build up in pipelines, pressure tanks, water heaters and
water softening equipment. These deposits restrict the flow of water and reduce water
pressure. More energy is required to pump water through clogged pipes and to heat
water if heating rods are coated with mineral deposits. This raises energy and water
costs.
 Water contaminated with iron and manganese often contains iron or manganese
bacteria. These bacteria feed on the minerals in the water. They do not cause health
problems, but do form a reddish-brown (iron) or brownish-black (manganese) slime in
toilet tanks and can clog water systems.

AIM
To determine the amount of manganese present in the given water sample.
PRINCIPLE
The concentration of manganese seldom exceeds a few mg/L. So, colorimetric methods are
applicable. The methods are (1) persulphate method, and (2) periodate method. Both methods
depend upon oxidation of manganese from its lower oxidation state to VII, where it forms the
highly coloured permanganate ion. The colour produced is directly proportional to the
concentration of manganese present over a considerable range of concentration in accordance
with Beer’s law. So it can be measured by eye or photometric means. Provisions must be
made to overcome the influence of chlorides. Manganese can also be determined by atomic
absorption spectrophotometry.
Persulphate oxidation of soluble manganous compounds to form permanganate is carried out

in the presence of silver nitrate. The resulting colour is stable for at least 24 h if excess
persulphate is present and organic matter is absent. Samples that have been exposed to air
may give low results due to the precipitation of MnO2. Adding one drop of 30% hydrogen
peroxide to the sample after the addition of special reagent re-dissolves precipitated
manganese.
Persulphate method is suitable because pre-treatment of samples is not required. Chloride

concentration is reduced by using mercuric sulphate. Persulphate oxidises manganese to
permanganate in the presence of silver nitrate as catalyst. The colour intensity is observed at a
wavelength of 525 nm in a spectrophotometer.
APPARATUS
o Colorimetric equipment; one of the following is required:

c) Spectrophotometer, for use at 525 nm, providing a light path of 1 cm or longer.
d) Nessler tubes, matched, 100 mL, tall form.
o Glassware like conical flasks, pipettes and glass beads.
REAGENTS
 Special reagent

 Ammonium persulphate
 Standard manganese solution
 Hydrogen peroxide 30%
Special Reagent: Dissolve 75 g HgSO4 in 400 mL conc. HNO3 and 200 mL distilled water.
Add 200 mL 85% phosphoric acid (H3PO4), and 35 mg silver nitrate (AgNO3). Dilute the
cooled solution to 1 L.
Ammonium Persulphate: (NH4)2S2O8, solid.
Standard Manganese Solution: Prepare a 0.1N potassium permanganate (KMnO4) solution

by dissolving 3.2 g KMnO4 in distilled water and making up to 1 L. Age for several weeks in
sunlight or heat for several hours near the boiling point, then filter through a fine fritted-glass
filter crucible and standardize against sodium oxalate.
Standard Manganese Solution (alternate): Dissolve 1g manganese metal (99.8% min.) in

10 mL redistilled HNO3. Dilute to 1000 mL with 1% (v/v) HCl; 1 mL ≡ 1 mg Mn. Dilute 10
mL to 200 mL with distilled water; 1 mL ≡ 0.05 mg Mn.
PROCEDURE
1. Take 50 mL of the sample in a conical flask. Add 50 mL distilled water to it.

2. Pipette 1, 2, 3, 4, and 8 mL of standard manganese solution to different flasks, and
dilute each to 100 mL using distilled water.
3. Add 5 mL special reagent to all the flasks.
4. Concentrate the solutions in all the flasks to about 90mL by boiling.
5. Add 1g ammonium persulphate to all the flasks, bring to boiling and boil for 1
minute.
6. Remove all the flasks from the heat source and let stand for 1 minute.
7. Then cool the flasks under the tap water.
8. Dilute the contents in all the flasks to 100mL with distilled water and mix. Pour the
contents into 100 mL Nessler tubes.
9. Match the colour of the sample with that of the colour standards. Note down the
concentration of Mn in µg.

10. If the spectrophotometer is used, one distilled water blank has to be prepared along
with the colour standards.
11. Measure the absorbance of each solution in a 1cm cell at 525 nm against the reference
blank prepared by treating distilled water.
12. Prepare the calibration graph taking meter reading along y-axis and concentration of
manganese (in µg) in colour standards on x-axis.
13. Keep the sample in the spectrophotometer and note down the meter reading.
14. Read off from the graph, the corresponding concentration of manganese in µg.
CALCULATION
OBSERVATIONS
Spectrophotometer Analysis
Volume of Mn Content from Concentration of Mn

No. Absorbance
Sample (mL) Graph (µg) (mg/L)
Visual Comparison
Volume of Concentration of Mn in Matching Concentration of Mn
No.
Sample (mL) Color Standard (µg) (mg/L)

RESULT
The amount of Mn present in the given sample is = .................................................. mg/L
QUESTIONS
1. In what oxidation state must the manganese be for colorimetric measurement?

2. In what oxidation state must the manganese be for colorimetric analysis? Why?
3. In the colorimetric determination of Mn:
a) What is the function of ammonium persulphate?
b) What is the function of Ag+?
c) What is the function of HgSO4?

DETERMINATION OF SULPHATE
INTRODUCTION
Sulphate (SO42–) is widely distributed in nature and may be present in natural waters in
concentrations ranging from a few to several thousand milligrams per liter. Mine drainage
wastes may contribute large amounts of SO42– through pyrite oxidation. Sodium and
magnesium sulphate exert a cathartic action.
 Excess sulphates in the form of Na2SO4 and MgSO4 should not be present in drinking
water as they cause cathartic action.
 In anaerobic decomposition of wastewater, sulphates are reduced to hydrogen
sulphide causing obnoxious odor and promote corrosion of sewers.
 Sulphates are reduced to sulphides in sludge digesters and may upset the biological
processes, if the sulphide concentration exceeds 200 mg/L.
AIM
To determine the sulphate content in the given water sample.
METHOD 1: GRAVIMETRIC METHOD WITH IGNITION OF RESIDUE
PRINCIPLE
Sulphate is precipitated in a hydrochloric acid (HCl) solution as barium sulphate (BaSO4) by

the addition of barium chloride (BaCl2). The precipitation is carried out near the boiling
temperature, and after a period of digestion the precipitate is filtered, washed with water until
free of Cl–, ignited or dried, and weighed as BaSO4.
APPARATUS
o Drying oven
o Desiccator
o Steam bath
o Analytical balance
o Ashless filter paper (Whatman filter paper No. 42)

o Muffle furnace
o Glassware like funnel, flask and pipette
REAGENTS
 Methyl red indicator solution

 Barium chloride solution.
 Silver nitrate–nitric acid reagent
Methyl Red Indicator Solution: Dissolve 100 mg methyl red sodium salt in distilled water
and dilute to 100 mL.
Hydrochloric Acid: 1+1 HCl.
Barium Chloride Solution: Dissolve 100 g BaCl2⋅2H2O in 1 L distilled water. Filter through
a membrane filter or hard- finish filter paper before use; 1 mL is capable of precipitating
approximately 40 mg SO42–.
Silver Nitrate-Nitric Acid Reagent: Dissolve 8.5 g AgNO3 and 0.5 mL concentrated HNO3
in 500 mL distilled water.
PROCEDURE
1. Take 250mL of the sample in a conical flask.

2. Adjust the acidity with HCl to 4.5 to 5 using a pH meter or the orange colour of
methyl red indicator.
3. Then add an additional 1 to 2 mL HCl.
4. Heat the solution to boiling and while stirring gently, add barium chloride solution
slowly until precipitation appears to be completed. Then add about 2 mL in excess.
5. Digest the precipitate at 80°C to 90°C preferably overnight but for not less than 2
hours.
6. Filter the contents in the flask through an ashless filter paper.
7. Wash the precipitate with small portion of warm distilled water until the washing is
free of chloride as indicated by testing with silver nitrate–nitric acid reagent.

8. Place the precipitate along with filter paper in a crucible after finding its empty weight
and dry it.
9. Keep the crucible in a muffle furnace and ignite at 800°C for 1 hour.
10. Cool in a desiccator and weigh.
11. Find weight of the barium sulphate precipitate.
CALCULATION
OBSERVATIONS
Weight of filter paper = ………………………. mg

Wt. of
Empty Crucible +
Volume of Weight of Residue Wt. of BaSO4 Concentration
No. Sample The Crucible After Precipitated of SO42-
(mL) + Filter Ignition + (mg) (mg/L)
Paper (mg) Filter Paper
(mg)
RESULT
The amount of sulphate present in the given sample = ……………… mg/L
METHOD 2: TURBIDIMETRIC METHOD
PRINCIPLE
Sulphate ion (SO42–) is precipitated in an acetic acid medium with barium chloride (BaCl2) so
as to form barium sulphate (BaSO4) crystals of uniform size. Light absorbance of the BaSO4
suspension is measured by a nephelometer or turbidimeter and the SO42– concentration is
determined by comparison of the reading with a standard curve.
APPARATUS
o Nephelometer or Turbidimeter
o Magnetic stirrer
REAGENTS
 Conditioning agent
 Barium chloride
 Standard sulphate solution
Conditioning Reagent: Mix 50 mL of glycerol with a solution containing 30 mL

concentrated HCl, 300 mL distilled water, 100 mL 95% ethyl or isopropyl alcohol and 75 g
NaCl.
Standard Sulphate Solution: Dilute 10.4 mL standard 0.02 N H2SO4 titrant to 100 mL with
distilled water or Dissolve 0.1479 g anhydrous Na2SO4 in distilled water and dilute to 1000
mL.
Barium Chloride: BaCl2, crystals, 20 to 30 mesh. In standardization, uniform turbidity is

produced with this mesh range.
HCl Solution: 1 mL of concentrated HCl + 9 mL of distilled water
PROCEDURE
1. Preparation of calibration curve: Prepare a series of standards by diluting 5, 7.5, 10,

12.5, 15, 17.5 and 20 mL of standard sulphate solution to 50 mL in volumetric flasks.
These solutions will have sulphate concentrations of 10, 15, 20, 25, 30, 35 and 40
mg/L, respectively.
2. Add 1 mL HCl solution and 1 mL of conditioning reagent and mix it well.
3. Dilute it to 50 mL with distilled water.
4. Add a spoon of barium chloride and mix it well.
5. Read the turbidity of each sample using nephelometer.
6. Prepare the calibration curve for standards (mg/L of sulphate vs turbidity).

7. Sample analysis: Take 20 mL of the sample (whose sulphate concentration is to be
determined) in a 50 mL volumetric flask and repeat the steps 2 to 5. Note down the
turbidity (A).
8. Blank correction: Take 20 mL of sample in a 50 mL volumetric flask, add 1 mL HCL
solution, add 1 mL of conditioning reagent, dilute to 50 mL and find its turbidity (B).
9. Therefore turbidity due to sulphate is A-B.
10. Read the sulphate concentration of the sample corresponding to the turbidity value
equal to A-B, directly from the calibration curve.
OBSERVATIONS
Sample No.
No. Turbidity (NTU)
or Description
RESULT
The amount of sulphate present in the given sample = ……………… mg/L
QUESTIONS
1. What is the purpose of digestion of the sample in the gravimetric analysis for
sulphates?
2. The water to be used for the preparation of cement concrete products should be free
from excess of sulphates and chlorides. Why?
3. List four precautions that must be observed to ensure an accurate gravimetric
determination of sulphate concentration.
4. What is the need of blank correction in turbidimetric analysis?

DETERMINATION OF SULPHIDE
INTRODUCTION
Sulphide often is present in groundwater, especially in hot springs. Its common presence in
wastewaters comes partly from the decomposition of organic matter, sometimes from
industrial wastes, but mostly from the bacterial reduction of sulphate. Water containing
hydrogen sulphide, commonly called sulphur water, has a distinctive "rotten egg" or swampy
odor. Hydrogen sulphide is a gas formed by the decay of organic matter such as plant
material. It is typically found in groundwater containing low levels of dissolved oxygen and a
pH less than 6.0. If the pH range of the water is higher (7.0-12.0), the water may contain
other forms of sulphur (sulphide or bisuphide).
 Sulphur problems occur less frequently in surface waters because flowing water is
aerated naturally so that the hydrogen sulphide reacts with oxygen and escapes as a
gas or settles as a solid.
 Hydrogen sulphide escaping into the air from sulphide-containing wastewater causes
odor nuisances. The threshold odor concentration of H2S in clean water is between
0.025 and 0.25 µg/L.
 Gaseous H2S is very toxic and has claimed the lives of numerous workers in sewers.
 At levels toxic to humans it interferes with the olfactory system, giving a false sense
of the safe absence of H2S.
 Sulphide attacks metals directly and indirectly has caused serious corrosion of
concrete sewers because it is oxidized biologically to H2SO4 on the pipe wall.
 Dissolved H2S is toxic to fish and other aquatic organisms.
AIM
To determine the sulphide content in the given water sample.
PRINCIPLE
Iodine reacts with sulphide in acid solution, oxidizing it to sulphur. A titration based on this
reaction is an accurate method for determining sulphides at concentration above 1mg/L, if

interferences are absent and if loss of H2S is avoided. This method is suitable for analyzing
samples freshly taken from wells or springs.
APPARATUS
o Burette
o Pipette
o Erlenmeyer flask.
REAGENTS
 Standard iodine solution (0.025N)
 Starch solution
Standard Iodine Solution (0.025 N): Dissolve 20 to 25 g KI in a little water and add 3.2 g
iodine. After iodine has dissolved, dilute to 1000 mL and standardize against 0.025 N
Na2S2O3, using starch solution as indicator.
Hydrochloric Acid: 6 N HCl.
Standard Sodium Thiosulphate Solution (0.025 N): Dissolve 6.205 g Na2S2O3⋅5H2O in

distilled water. Add 1.5 mL 6 N NaOH or 0.4 g solid NaOH and dilute to 1000 mL.
Standardize with bi-iodate solution.
PROCEDURE
1. Measure from a burette 10 mL of iodine into a 500 mL flask.

2. Add distilled water and bring the volume to 20 mL.
3. Add 2 mL of 6 N HCl.
4. Pipette 200 mL sample into the flask, discharging the sample under the surface of
solution.
5. If the iodine colour disappears, add more iodine so that the colour remains.
6. Titrate with sodium thiosulphate solution, adding a few drops of starch solution, as
the end point is approached and continuing until the blue colour disappears.
CALCULATION
where, A = Volume of 0.025 N iodine solution used (mL)
B= Volume of 0.025 N sodium thiosulphate solution used (mL)
OBSERVATIONS

Indicator: Starch
Volume of Sodium Thiosulphate
Volume of Iodine Solution (mL)
Volume (mL)
of Volume Volume
No. Initial Final Initial Final
Sample of Iodine of
(mL) Burette Burette Burette Burette
Solution Na2S2O3
Reading Reading Reading Reading
Used ‘A’ Used ‘B’
RESULT
The amount of sulphide present in the given sample = ……………… mg/L
QUESTIONS
1. Explain the significance of the determination of sulphide concentration in

environmental engineering.
2. Between sulphate and sulphide, which one is more toxic to environment? Why?

TEST FOR COLIFORMS IN WATER
INTRODUCTION
Water pollution caused by fecal contamination is a serious problem due to the potential for
contracting diseases from pathogens (disease causing organisms). Frequently, concentrations
of pathogens from fecal contamination are small, and the number of different possible
pathogens is large. As a result, it is not practical to test for pathogens in every water sample
collected. Instead, the presence of pathogens is determined with indirect evidence by testing
for an "indicator" organism such as coliform bacteria. Coliforms come from the same sources
as pathogenic organisms. Coliforms are relatively easy to identify, are usually present in
larger numbers than more dangerous pathogens, and respond to the environment, wastewater
treatment, and water treatment similarly to many pathogens. As a result, testing for coliform
bacteria can be a reasonable indication of whether other pathogenic bacteria are present.
The most basic test for bacterial contamination of a water supply is the test for total coliform
bacteria. Total coliform counts give a general indication of the sanitary condition of a water
supply.
 Total coliforms include bacteria that are found in the soil, in water that has been
influenced by surface water, and in human or animal waste.
 Fecal coliforms are the group of the total coliforms that are considered to be present
specifically in the gut and feces of warm-blooded animals. Because the origins of fecal
coliforms are more specific than the origins of the more general total coliform group of
bacteria, fecal coliforms are considered a more accurate indication of animal or human
waste than the total coliforms.
 Escherichia coli (E. coli) is the major species in the fecal coliform group. Of the five
general groups of bacteria that comprise the total coliforms, only E. coli is generally not
found growing and reproducing in the environment. Consequently, E. coli is considered
to be the species of coliform bacteria that is the best indicator of fecal pollution and the
possible presence of pathogens.

 Most coliform bacteria do not cause disease. However, some rare strains of E. coli,
particularly the strain 0157:H7, can cause serious illness. Outbreaks of disease caused
by E. coli 0157:H7 have generated much public concern about this organism. E. coli
0157:H7 has been found in cattle, chickens, pigs, and sheep. Most of the reported
human cases have been due to eating under cooked hamburger. Cases of E. coli
0157:H7 infection caused by contaminated drinking water supplies are rare.
 If coliform bacteria are present in drinking water, the risk of contracting a water-borne
illness is increased. Although total coliforms can come from sources other than fecal
matter, a positive total coliform sample should be considered an indication of pollution.
Positive fecal coliform results, especially positive E. Coli results, should be considered
indication of fecal pollution.
AIM
To determine the most Probable number (MPN) index of coliforms and E coli or (B coli)
organisms in the given sample (s) of water by the Multiple Tube Fermentation Technique.
PRINCIPLE
The coliform group of bacteria has been accepted as the indicator organisms for fecal
pollution of water .The E coli groups of organisms further confirms the presence of fecal
matter in the water tested. Since water born infections are essentially forced by sewage or
fecal contamination, the detection of coliform organisms in water which indicate excretal
contamination is of supreme importance.
The coliform group comprises the entire aerobic and facultative anaerobic gram negative, non
spore forming, rod-shaped bacteria which ferment lactose with gas formation within 48hours
at 350C.
The E coli group belongs to the coliform of fecal origin. They ferment lactose with gas
formation within 24 hours at 440C.
The water is considered to be safe when these two organisms are absent. This should be
conducted as routine experiments in all the water treatment plants to check the efficiency of
disinfection.
APPARATUS
o Incubator
o Test tubes
o Platinum Loops etc.
REAGENTS
 Lauryl Tryptose Broth

 Brilliant Green Lactose Bile Broth
 EC Medium
MEDIA PREPARATION
Lauryl Tryptose Broth: Dissolve 20 g tryptose, 5 g lactose, 2.75 g dipotassium hydrogen

phosphate (K2HPO4), 2.75 g potassium dihydrogen phosphate (KH2PO4), 5 g NaCl, 0.1 g
sodium lauryl sulphate in 1 L distilled water. 10 mL of this medium is put into each of test
tube. If 50 mL quantities are to be tested, 50 mL of the broth should be put into tubes or
bottles of suitable size. Sterilize at 10 psi (0.75kg /cm2) pressure for 15 minutes on 3
successive days.
Brilliant Green Lactose Bile Broth: Dissolve 10 g peptone in 500 mL with distilled water.
Steam to dissolve and add 20 g dehydrated ox-gall and 200 mL distilled water.
The ox-gall solution should be adjusted to a pH between 7 and 7.5. Make up with distilled
water to approximately 975mL. Add 10g lactose, adjust pH to 7.4 add 13mL of 3.1 percent
solution of brilliant green in distilled water and make up to 1000 mL.
Distribute in 5mL quantities in fermentation tubes and sterilize in the autoclave at 10 psi
(0.75 kg/cm2), pressure for 10minutes on three successive days. The pH which may be
determined by potentiometric method after sterilization shouldn’t be less than 7.1 and not
more than 7.4.
EC Medium: Dissolve 20 g tryptose or trypti case, 5 g lactose, 15 g bile salts (mixture or bile
salts no.3), 4 g dipotassium hydrogen phosphate (K2HPO4), 1.5 g potassium dihydrogen
phosphate (KH2PO4), 5 g NaCl in 1 L distilled water. Prior to sterilization, dispense in
fermentation tubes with sufficient medium (10 mL) to cover the inverted Durham tubes at
least partially after sterilization. pH should be 6.9 after sterilization.
PROCEDURE
1. Collect the sample in sterilized bottles intended for bacteriological analysis.

2. Prepare the sterilized media necessary for the bacteriological test and keep them ready
in test tubes containing Durham tubes.
3. Inoculate the sample in logarithmic dilutions i.e., 10, 1 and 0.1mL in 5 tubes each of
Lauryl tryptose broth under complete aseptic conditions.
4. Incubate all the tubes at 350C.
5. After 24 hours examine the tubes for gas formation.
6. The tubes containing the gas are marked positive and are taken out of the incubator
for further analysis. The remaining tubes are further incubated at 350C for another 24
hours. This is the presumptive test for coliform organisms.
7. Take one or two loop full of the liquid from the positive Lauryl Tryptose tubes and
inoculate in Brilliant Green Lactose bile broth tubes and incubate the tubes for 24
hours at 350C.
8. At the end of 24 hours examine the tubes for gas formation .The presence of gas
confirms the presence of coliform organisms. The negative tubes are further incubated
for another 24 hours and the presence or absence of gas is noted. This is the
confirmatory test for coliform organisms.
9. One or two loop full of the liquid from the positive Lauryl Tryptose tubes are
transferred into the sterilized EC medium tubes. They are incubated for 24 hours at
44oC.
10. The gas production after 24 hours, confirms the presence of E coli organisms or fecal
coliforms.
CALCULATION
Tabulate the results (in the adjoining) tabular column and find out the corresponding MPN
index from the MPN table.

OBSERVATIONS
Volume of sample incubated in mL Total number

Sample No. Date and Date and of positive
or time of time of tubes after
10 10 10 10 10 1 1 1 1 1 0.1 0.1 0.1 0.1 0.1
Description incubation observation confirmatory
MPN Test for Remarks
test
24 hours
presumptive
test
48 hours
presumptive
test
24 hours
confirmatory
Coliform test
organisms 48 hours
confirmatory
test
24 hours
E coli confirmatory
test

RESULT
MPN of coliform organisms =…………………./100mL of the sample

MPN of E coli or B coli organisms =…………………./100mL of the sample
QUESTIONS
1. What are E coli? Are they harmful to human beings? Why is their presence tested in
the waters to be supplied for domestic consumption?
2. What is coliform index?
3. Define MPN.

DETERMINATION OF OPTIMUM COAGULANT DOSAGE
INTRODUCTION
Coagulation: The process in which chemicals are added to water, causing a reduction of the
forces tending to keep particles apart. Particles in source water are in a stable condition. The
purpose of coagulation is to destabilise particles and enable them to become attached to other
particles so that they may be removed in subsequent processes. Particulates in source waters
that contribute to colour and turbidity are mainly clays, silts, viruses, bacteria, fulvic and
humic acids, minerals (including asbestos, silicates, silica, and radioactive particles), and
organic particulates. At pH > 4.0, particles or molecules are generally negatively charged.
The coagulation process physically occurs in a rapid mixing process.
Flocculation: The agglomeration of small particles and colloids to form settleable or

filterable particles (flocs). Flocculation begins immediately after destabilisation in the zone of
decaying mixing energy following rapid mixing, or as a result of the turbulence of
transporting flow. In some instances, this incidental flocculation may be an adequate
flocculation process. A separate flocculation process is most often included in the treatment
train to enhance contact of destabilised particles and to build floc particles of optimum size,
density, and strength.
 Coagulation removes not only turbidity, but also colour, microorganisms, algae,
phosphate, taste and odour.

AIM
To find the optimum amount of coagulant required to treat turbid water.
PRINCIPLE
Salts of aluminium and iron are commonly used as coagulants in water and wastewater
treatment. When a salt of aluminium and iron is added to water, it dissociates to yield
trivalent ions, which hydrate to form aquometal complexes Al(H2O)63+ and Fe(H2O)63+.
These complexes then pass through a series of hydrolytic reactions in which H2O molecules
in the hydration shell are replaced by OH- ions to form a variety of soluble species such as
Al(OH)2+ and Fe(OH)2+. These products are quite effective as coagulants as they adsorb very
strongly onto the surface of most negative colloids.
Jar test is a simple method used to determine this optimum coagulant dose
required. The jar test device consists of a number of stirrers (4 to 6) provided with
paddles. The paddles can be rotated with varying speed with the help of a motor
and regulator. Samples will be taken in jars or beakers and varying dose of
coagulant will be added simultaneously to all the jars. The paddles will be rotated
at 100 rpm for 1 minute and at 40 rpm for 9 minutes, corresponding to the flash
mixing and slow mixing in the flocculator of the treatment plant. After 10 minutes
settling, supernatant will be taken carefully from all the jars to measure turbidity.
The dose, which gives the least turbidity, is taken as the optimum coagulant dose.
APPARATUS
o Jar test apparatus

o Glass beakers
o Pipette
o Nephelometer
o pH meter
REAGENTS
 Alum solution
 Lime

 Acid/alkali
REAGENT PREPARATION
Alum Solution: Dissolve 10 g of alum in 100 mL of distilled water.
PROCEDURE
1. Take 1000 mL of given sample in 6 beakers.

2. Find the pH of the sample and adjust it to 6 to 8.5.
3. Add various dosages of alum in each one of the beakers.
4. Insert the paddle of the jar testing apparatus inside the beakers and start it.
5. Initially maintain a speed such that the paddles rotate at an angular velocity of 100
rpm for a time of 1 minute.
6. Adjust the speed such that the paddles rotate at 40 rpm for a time of 9 minutes.
7. Allow the beakers to settle down for 10 minutes.
8. Make an observation as of which of the 6 beakers is clearer. Also measure the
turbidity of each beaker using a turbidity meter and tabulate your results.
9. Plot a graph “Residual turbidity” Vs “Coagulant dosage” to determine the optimum
coagulant dosage.
OBSERVATIONS
Initial Turbidity of the sample: ………………………… NTU

pH of the solution: ……………………..
Volume of Sample Dose of Alum Residual
No.
(mL) mL mg/L Turbidity (NTU)

RESULT
Optimum dosage of coagulant is = ……………mg/L
QUESTIONS
1. List various coagulants used for water treatment? Why is alum preferred to other
coagulants?
2. What is the difference between coagulation and flocculation?
3. What are coagulant aids? Give examples.
4. Write the significance of pH in coagulation using alum.
5. What factors affect the sedimentation of a discrete particle setting in a quiescent
liquid?
6. For efficient coagulation the water must be alkaline. Why?
7. Explain various coagulation mechanisms in detail? What is the coagulation
mechanism of the laboratory experiment that you were carried out? Why?
8. Explain various flocculation mechanisms.
9. The ideal depth of immersion of paddles in jar test is 2/3rd of the total depth. Why?
10. Explain the coagulation activities of copperas.
11. What is the mechanism behind sedimentation or delta formation occuring at
estuaries (Estuary is the portion of the river where it meets sea)?

DETERMINATION OF AVAILABLE CHLORINE IN BLEACHING
POWDER AND TEST FOR RESIDUAL CHLORINE
INTRODUCTION
The chlorination of water supplies and polluted waters serves primarily to destroy or
deactivate disease-producing microorganisms. A secondary benefit, particularly in treating
drinking water, is the overall improvement in water quality resulting from the reaction of
chlorine with ammonia, iron, manganese, sulphide, and some organic substances.
Chlorination may produce adverse effects. Taste and odour characteristics of phenols and
other organic compounds present in a water supply may be intensified. Potentially
carcinogenic chloro-organic compounds such as chloroform may be formed. Combined
chlorine formed on chlorination of ammonia- or amine-bearing waters adversely affects some
aquatic life. To fulfil the primary purpose of chlorination and to minimize any adverse
effects, it is essential that proper testing procedures be used with a foreknowledge of the
limitations of the analytical determination.
Bleaching powder is commonly used as a disinfectant. The chlorine present in the bleaching
powder gets reduced with time. So, to find the exact quantity of bleaching powder required,
the amount of available chlorine in the sample must be found out.
 Active chlorine (free and combined) should be determined at each stage in the
treatment process of drinking water and in the water mains in order to guarantee
bacteriologically impeccable water.
 Active chlorine should present in drinking water within the range of 0.1 to 0.2 mg/L.
However, excessive chlorine content may give out bad odour and may change even
taste of water.
 Chlorine is said to be carcinogenous. Hence, except during epidemics ‘super
chlorination’ is not to be done.
 Chlorine residuals determination is used to control chlorination of domestic and
industrial wastewater.

 Determination of chlorine residual in water distribution is useful to find the source of
contamination or leakage points, so as to supply wholesome water to the consumer.
AIM
The aims of the experiment are:
a) To determine the amount of available chlorine in the bleaching powder by using

iodometric method and,
b) To find the chlorine dosage for the given sample of water using orthotolodine method.
PRINCIPLE
Iodometric Method: Chlorine will liberate free iodine from potassium iodide (KI) solutions
at pH 8 or less (Eq. 1). The liberated iodine is titrated with a standard solution of sodium
thiosulphate (Na2S2O3) with starch as the indicator (Eq. 2). Titrate at pH 3 to 4 because the
reaction is not stoichiometric at neutral pH due to partial oxidation of thiosulphate to
sulphate. The disappearance of blue colour indicates the completion of reaction with free
iodine converted to iodide.
(1)
(2)
Orthotolodine Method: It measures both free and combined available chlorine. To obtain
the correct colour development with chlorine and orthotolodine, (a) the solution must be at
pH 1.3 or lower during the contact period (b) the ratio by weight of orthotolodine to chlorine
must be at least 3:1 and, (c) the chlorine concentration must not exceed 10 mg/L. The
orthotolodine reacts with residual chlorine and give yellow colour to the sample. The
intensity of colour formation is proportional to the amount of residual chlorine present.
Usually permanent colour standards representing different values of chlorine residuals will be
prepared and kept ready for comparison.
APPARATUS
o Burette
o Pipettes
o Erlenmeyer flasks
o Mortar and pestle
o Chloroscope etc.
REAGENTS
 Concentrated glacial acetic acid

 Potassium iodide
 Starch indicator
 Iodine solution (0.025N)
 Orthotolodine reagent
Standard Sodium Thiosulfate Titrant: Dissolve 6.205 g Na2S2O3⋅5H2O in distilled water.

Add 1.5 mL 6N NaOH or 0.4 g solid NaOH and dilute to 1000 mL. Standardize with bi-
iodate solution.
Orthotolodine Reagent: Dissolve 1.35 g of orthotolodine dihydrochloride in 500 mL

distilled water. Add this solution with constant stirring to a mixture of 350 mL distilled water
and 150 mL concentrated HCl. Store the solution in brown bottle.
PROCEDURE
A. Available Chlorine in Bleaching Powder using Iodometric Method

1. Dissolve 1g bleaching powder in 1 litre of distilled water in a volumetric flask, and
stopper the container. (This can be done by first making a paste of the bleaching
powder with mortar and pestle.).
2. Place 5 mL acetic acid in an Erlenmeyer flask and add about 1g potassium iodide
crystals. Pour 25 mL of bleaching powder solution prepared above and mix with a
stirring rod.
3. Titrate with 0.025 N sodium thiosulphate solution until a pale yellow colour is
obtained. (Deep yellow changes to pale yellow.)

4. Add 1mL of starch solution and titrate until the blue colour disappears.
5. Note down the volume of sodium thiosulphate solution added (A).
6. Take a volume of distilled water corresponding to the sample used.
7. Add 5 mL acetic acid, 1g potassium iodide and 1 mL starch solution.
8. If blue colour occurs, titrate with 0.025 N sodium thiosulphate solution until the blue
colour disappears.
9. Record the volume of sodium thiosulphate solution added (B1).
10. If no blue colour occurs, titrate with 0.025 N iodine solution until a blue colour
appears. Note down the volume of iodine (B2).
11. Then, titrate with 0.025 N sodium thiosulphate solution till the blue colour disappears.
Record the volume of sodium thiosulphate solution added (B3). Note down the
difference between the volume of iodine solution and sodium thiosulphate as B4
(B4=B2 – B3).
Note: Blank titration is necessary to take care of (a) the oxidising or reducing reagents’
impurities (b) the iodine bound to starch at the end point.
B. Determination of Chlorine Dosage for the Given Water Sample using the above
Bleaching Powder Solution
1. Measure 200 mL of the given sample of water into 5 flasks of ample capacity to
permit mixing.
2. Add suitable increasing amounts of bleaching powder solution to the successive
flasks in such a way that the chlorine added will be 0.1 mg/L, 0.2 mg/L, 0.5 mg/L, 1
mg/L and 1.5 mg/L as mentioned in step 4 below.
3. Mix while chlorine solution is being added to the sample.
4. Dose the portions of the sample according to the staggered schedule that will permit
the determination of chlorine residuals at 30 min contact time.
5. At the end of contact period, pour the solution from the flask into the middle chamber
of the cell of the chloroscope. Add 5 drops of orthotolodine solution and mix well
with the plunger. Fill the outer chambers with unchlorinated water sample. Leave for
5 minutes.
6. Put the glass disc which indicates the dose of chlorine on its top, in front of the outer
chamber.
7. Compare the colour of the middle chamber with the colour of the glass disc.

8. The disc which matches with the colour of the sample will give the residual chlorine
present in the water.
CALCULATION
A. Iodometric Method
1000 mL of bleaching powder contains = 1000 x .................................. mg of chlorine
i.e. 1000 mg of bleaching powder contains ................................ mg of chlorine
Percentage of chlorine available in the bleaching powder (X) = ...............................................
B. Orthotolodine Method
Select the lowest dose which gives 0.2 mg/L of residual chlorine as chlorine dosage (Y).
Chlorine demand = Amount of chlorine added – 0.2 mg/L residual chlorine
OBSERVATIONS
A. Iodometric Method
Bleaching powder solution × Standard sodium thiosulphate solution (0.025 N)

Burette solution: Sodium Thiosulphate
Indicator: Starch
Volume of Bleaching Burette Reading (mL) Volume of Sodium
No.
Powder (mL) Initial Final Thiosulphate ‘A’ (mL)

Distilled water × Standard sodium thiosulphate solution (0.025 N)
Indicator: Starch
Volume of Distilled Water Burette Reading (mL) Volume of Sodium
No.
(mL) Initial Final Thiosulphate ‘B1’(mL)
Distilled water × Standard iodine solution (0.025N)

Volume of iodine added (B2) = .................... mL
Indicator: Starch
Volume of Distilled Water Burette Reading (mL) Volume of Sodium
No.
(mL) Initial Final Thiosulphate ‘B3’(mL)
B. Orthotolodine Method
No. Chlorine Added Bleaching Powder Residual Chlorine
(mg/L) Added (mL) (mg/L)

RESULT
The amount of available chlorine in the given bleaching powder is = ......................... %
Chlorine demand of the water sample = ............................... mg/L
Chlorine dosage of the water sample = ...................................mg/L
Bleaching powder required to treat 1 L of water = ..........................mg
QUESTIONS
1. What is disinfection? Differentiate between disinfection and sterilisation?

2. Why do we prefer chlorination over other methods of disinfection?
3. What is electro-katadyn process?
4. What are the different factors that affect the degree of disinfection?
5. Differentiate between free residual chlorine and combined residual chlorine.
6. The dosage of chlorine should be proper. Why?
7. What is the use of acetic acid addition in iodometric method?
8. Discuss the effect of pH of water and organic matter of water on efficiency of
disinfection by chlorine.
9. By use of appropriate equilibrium equations show why the addition of chlorine tends
to decrease the pH of water, while hypochlorite tends to increase the pH.
10. Explain various chlorination methods in detail.


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INTRODUCTION

Physical Characteristics: Turbidity, Colour, Taste, Odour, Temperature, Electrical

Chemical Characteristics: Solids, pH, Hardness, Alkalinity, Chloride, Metals, Dissolved

1 Environmental Engineering Laboratory Manual – NIT Calicut, Kerala

TYPES OF WATER ANALYSIS

Gravimetric Analysis: Gravimetric analysis describes a set of methods in analytical

Volumetric Analysis (Titrimetry): Volumetric analysis is a widely-used quantitative

Colourimetric Analysis: It is a method of determining the concentration of a chemical

A colourimeter is a device used to test the concentration of a solution by measuring its

2 Environmental Engineering Laboratory Manual – NIT Calicut, Kerala

3 Environmental Engineering Laboratory Manual – NIT Calicut, Kerala

DOs AND DON’Ts IN THE LABORATORY

 Do thoroughly clean the glassware before and after use.

4 Environmental Engineering Laboratory Manual – NIT Calicut, Kerala

To determine the pH value of the given water sample.

5 Environmental Engineering Laboratory Manual – NIT Calicut, Kerala

o pH meter with electrode

(a) Using Universal Indicator

1. 10 mL of sample is taken in a cuvette.

(b) Using pH Papers

1. Dip the pH paper in the sample.

(c) Using pH Meter

1. Follow the manufacturer’s operating instructions.

Temperature of the sample = .................................... oC

7 Environmental Engineering Laboratory Manual – NIT Calicut, Kerala

The pH of the given sample is = ..................................................

1. A decrease in pH of 1 unit represents how much of an increase in hydrogen ion

8 Environmental Engineering Laboratory Manual – NIT Calicut, Kerala

 Turbidity is objectionable because of (a) aesthetic considerations and (b) engineering

9 Environmental Engineering Laboratory Manual – NIT Calicut, Kerala

To determine the turbidity of the given water sample.

o Nephelometric turbidity meter

Stock Primary Standard Formazin Suspension: Dissolve 1 g hydrazine sulfate,

1. Switch on the instrument.

No. Sample No. Turbidity (NTU)

The turbidity of the given sample is = .................................................. NTU

1. Where do you find the adverse effects of turbidity in environmental engineering?

12 Environmental Engineering Laboratory Manual – NIT Calicut, Kerala

13 Environmental Engineering Laboratory Manual – NIT Calicut, Kerala

(a) Total solids

14 Environmental Engineering Laboratory Manual – NIT Calicut, Kerala

o Porcelain evaporating dishes of 150–200 mL capacity

(a) Total Solids

15 Environmental Engineering Laboratory Manual – NIT Calicut, Kerala

(b) Total Fixed Solids

(c) Total Dissolved Solids

(d) Total Suspended Solids = Total solids – Total dissolved solids.

(e) Total Volatile Solids = Total solids – Total fixed solids.

(f) Fixed Dissolved Solids

(j) Settleable Solids by Volume

17 Environmental Engineering Laboratory Manual – NIT Calicut, Kerala

Item Sample 1 Sample 2 Sample 3

Volume of Sample (mL)

Empty Wt. of Dish (W1; mg)

Wt. of Dish + Total Solids ( W2; mg)

Total Solids (mg/L)

Total Fixed Solids

Item Sample 1 Sample 2 Sample 3

Volume of Sample (mL)

Empty Wt. of Dish (W1; mg)

Wt. of Dish + Total Fixed Solids ( W3; mg)

Total Fixed Solids (mg/L)