Env Lab Manual 2020

ENVIRONMENTAL ENGINEERING LAB MANUAL CVL77
INSTRUCTIONS
1. This laboratory manual is ONLY for reference for those using the Environmental
Engineering laboratory.
2. Discussion after each experiment should be based on the following points:
(a) Limit prescribed for that constituent in drinking water standards.

(b) The suitability of the sample for drinking purpose with respect to that particular
constituent.
3. To Note:
(a) In water and wastewater analysis, the results are usually reported in terms of mg/l of
some particular ion, element or compound. It is most convenient to have the
standard titrating agent of such strength, that 1mL is equivalent to 1mg of material
being measured. Thus 1 litre of the standard solution is usually equivalent to 1g of
the standard substance.
(b) Rules listed by Worthing and Geffner are to be followed while plotting graphs.
4. Users may refer the following for writing the discussion after each experiment:
(a) “Standard Methods for the Examination of Water and Waste Water”, American
Public Health Association, 1015, 15th Street, N.W., Washington D.C., 2005.
(b) “Chemistry for Environmental Engineers”, Sawyer and McCarty, Tata Mc-Graw
Hill.
(c) “Manual of Standards of Quality for Drinking Water Supplies”, Indian Council
of Medical Research, New Delhi.
(d) “International Standards for Drinking Water” — World Health Organisation.
(e) “IS 2490 - 1981, IS 3306 - 1974, IS 3307 - 1977, IS 7968 - 1976, IS 2296 – 1974”,
Bureau of Indian Standards, New Delhi.
DOS AND DONOT’S IN THE LABORATORY
1. Do thoroughly clean the glassware before and after use.

2. Do handle the glassware carefully.
3. Do not handle chemicals with bare hands.
4. 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.
5. Do not add water to acids. Do always add acid to water.
Experiment Content Hours COs

1 Determination of Solids in Sewage: Total Solids, Suspended
CO1
Solids, Dissolved Solids, Volatile Solids, Fixed Solids, Settle-able 10
CO4
Solids.
2 Electrical conductivity. Determination of Chlorides and Sulphates.
3 Determination of Alkalinity, Acidity and pH.
4 Determination of Calcium, Magnesium and Total Hardness.
5 Determination of Dissolved Oxygen. Determination of BOD.
6 Determination of COD.
CO2
7 Determination of percentage of available chlorine in bleaching
20 CO3
powder,
CO4
8 Residual Chlorine and Chlorine Demand.
9 Jar Test for Optimum Dosage of Alum, Turbidity determination by
Nephelometer.
10 Determination of Iron. Phenanthroline method.
11 Determination of Fluorides SPANDS Method.
12 MPN Determination
CO5
13 Determination Nitrates by spectrophotometer. 10
CO6
14 Determination of sodium and potassium by flame photometer
REFERENCES
1. Manual of Water and Wastewater Analysis – NEERI Publication.
2. Standard Methods for Examination of Water and Wastewater (1995), American Publication
Association, Water Pollution Control Federation, American Water Works Association,
Washington DC.
3. IS Standards : 2490-1974, 3360-1974, 3307-1974. 4. Chemistry for Environment
Engineering. Sawyer and Mc Carthy,
Name & Signature of Faculty In-charge Head of the Department

Preview
Concurrent with the evaluation of environmental engineering practice, analytical

methods have been developed to obtain the factual information required for the
resolution and solution of problems. Environmental engineering is perhaps most
demanding in this respect, for it requires the use of not only the conventional
measuring devices employed by engineers but, in addition many of the
techniques and methods of measurement used by chemists, physicists and some
of those.
The objective of sampling is to collect representative sample. Representative

sample by means a sample in which relative proportions or concentration of all
pertinent components will be the same as in the material being sampled. Samples
ought to be handled in such a way that no significant changes in composition
occur before the tests are made. The sample volume shall optimal small enough
that it can be transported and large enough for analytical purposes. Grab samples
are single collected at a specific spot at a site over a short period of time.
Composite samples provide a more representative sampling and can be obtained
by combining portions of multiple grab samples or by using specially designed
automatic sampling devices. Integrated samples are obtained by analyzing
mixtures of grab samples collected from different points simultaneously. The
type of sample containers used is also of utmost importance. Containers typically
are made of plastic or glass, but one material may be preferred over the other.
Because of variability from analytical and sampling procedures (i.e., population
variability), a single sample is often insufficient to reach any reasonable desired
level of confidence. If an overall standard deviation (i.e. the standard deviation of
combined sampling and analysis) is known, the required number of samples for
water may be estimated.
Complete and unequivocal preservation of samples, whether domestic

wastewater, industrial wastes, or natural waters, is a practical impossibility
because complete stability for every constituent never can be achieved. At best,
preservation techniques only retard chemical and biological changes that
inevitably continue after sample collection. If immediate analysis is not possible,
preferably store at 4°C. No single method of preservation is entirely satisfactory;
choose the preservative with due regard to the determinations to be made.
STANDARDS FOR DRINKING WATER

Sl.No Characteristics Acceptable cause for Remarks
Rejection
1 Turbidity (NTU) 1 10 Consumer acceptance decreases
2 Colour( Units on platinum cobalt 5 25
scale Hazen Unit)
3 PH 7.0 to 8.5 < 6.5 or > 9.2 Water will affect the mucous
membrane & / water supply
system
4 Total dissolved solids(mg/l) 500 2000 Consumer acceptance decreases
laxative effect upon people who
are not accustomed to it. May
cause gastro –intestinal irritation
5 Total hardness (as CaCo3) (mg/l) 200 600 Encrustation in water supply
structure and adverse effects on
domestic use / scale formation
6 Chlorides ( as CI ) (mg/l) 200 1000 Taste, palatability and corrosion
are affected
7 Sulphates ( as SO4) (mg/l) 200 400 Cause Gastro –intestinal irritation
8 Fluorides ( as F ) (mg/l) 1.0 1.5 Results in dental / skeletal
fluorosis
9 Nitrates ( as NO3) (mg/l) 45 100 May cause Methaemoglobineamia
/ Blue baby disease
10 Calcium (as Ca ) (mg/l) 75 200 Encrustation in water supply
11 Magnesium ( as Mg) (mg/l) <30 150 structure and adverse effects on
domestic use
12 Iron ( as Fe ) (mg/l) 0.1 1.0 Taste and appearance are affected
and promotes iron bacteria &
adverse effect on domestic user &
water structures
13 Alkalinity (mg/l) 200 600 Water will affect the mucous
membrane & / water supply
system, taste becomes unpleasant.
14 Arsenic ( as As ) (mg/l) 0.01 0.05 Water becomes toxic
15 Pesticides ( total, mg /l) Absent 0.001
16 Residual free chlorine (mg/l) 0.2 To be applicable when water is
chlorinated. Tested at consumer
end. When protection against viral
infection is required, it should be
min.0.5 mg/l
17 Bacteria (MPN /100ml) 0 10 Results in diarrhea, decentry
Experiment No.: 01
SOLIDS IN WASTE WATER
Aim
Determination of Solids in sewage: Total Solids, Suspended Solids, Dissolved Solids,

Volatile Solids, Fixed Solids and Settleable Solids.
Apparatus
China dish of 150–200 ml capacity, Muffle furnace, hot air oven, digital balance, filter paper,
Imhoff cone, conical flask, Desiccators.
Theory
One of the most important characteristics of sewage is the solid content. Sewage normally
contains 99 % water and 1% solids. Solids test is an important parameter as it is used in:
1. Design of biological units and water pollution control methods, which depend on the
concentration of organic solids.
2. In Grit chambers and sedimentation tank design which depends on the quantity of
inorganic and organic settleable solids respectively.
3. Dissolved inorganic solids are to be considered in the design of land treatment of
Sewage.
Classification of Sewage Solids:
Total solids: Analytically the total solids content of a wastewater is defined as all the matter
that remains as residue on evaporation at 103°C and subsequent drying. These solids are very
important as they indicate the strength of sewage and amount of treatment required.
Total solids in wastewater exist in 3 forms:
Dissolved solids, Colloidal solids and Suspended solids.
Dissolved solids are those which remain dissolved in sewage just as salt in water and are
generally organic in nature causing putrefaction and thus increasing the strength of sewage
and creating nuisance if disposed off untreated. Colloidal solids are finely divided solids
remaining either in solution or in suspension.
Suspended solids are those, which are in suspension and are floating in sewage.
These are further classified into:
 Settleable solids, which settle down at the bottom of an Imhoff cone and are an
approximate measure of the quality of sludge that will be removed by sedimentation.
 Non settleable solids are those, colloidal in nature and can be removed by Chemical
treatment.
1
Fixed solids: Fixed solids are also called inorganic solids consisting of minerals and salts,
which include sand, gravel, debris, dissolved salts etc. They constitute about 55°/o of the total
solids and are not harmful in sewage treatment operations and require only mechanical
appliances such as screens for their removal.
Volatile solids: They are also called organic solids or ignited solids, consisting of organic
matter such as carbohydrates, fats, oils etc and constitute about 45% of the total solids and
commonly used in the analysis of wastewater sludge's to measure their biological stability.
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 = WT = (W2 – W1) mg. [W2 and W1 should be expressed in mg.]
b] Total Fixed Solids
1. Keep the same dish (WT) 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 desiccator for final cooling in a dry atmosphere.
3. Weigh the dish as soon as it has cooled (W3).
4. Weight of total fixed residue = WF = (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 Whatmann
filter paper (No 44).
2. Heat and collect the filtrate in a previously prepared and weighed evaporating dish at
103 °C for 1 to 1.5 hours in a hot air oven and weigh the residue.
3. Allow the dish to cool briefly in air before placing it, while still warm in a desiccator to
complete cooling in a dry atmosphere.
4. Weigh the dish as soon as it has completely cooled (W5).
5. Weight of total dissolved solids = WD = (W5 – W4) mg.
Where,
W4 = Weight of empty evaporating dish in mg.
2
W5 = Weight of empty evaporating dish in mg + Residue left after evaporating the

filtrate in mg.
d] Total Suspended Solids = Ws = Total Solids – Total Dissolved Solids.
e] Total Volatile Solids = WV = Total Solids – Total Fixed Solids.
f] Settleable Solids (by volume) [WSS]
1. Fill an imhoff cone (lit capacity, graduated up to 40ml) to the litre mark with a
thoroughly mixed sample.
2. Allow to settle for 45 minutes.
3. Gently stir the sides of the cone with a rod a few times to ensure that all solids
adhering to the sides is loosened.
4. Allow to settle for further 15 minutes.
5. Record the volume of settleable matter in the cone as ml/l.
Observations, Calculations and Results
Parameter Label Unit Result

Volume of sample taken V ml
Total solids
Weight of clean empty evaporating dish W1 mg
Weight of china dish after it has completely W2 mg
cooled with residue of sewage sample oven
dried at 103°C
Total Solids WT = W2 – W1 mg
Total Solids (per ml of sample) [WT x 103] / V mg/l
Total Fixed Solids

Total Fixed Solids WF = W3 – W1 mg
Total Fixed Solids (per ml of sample) [WF x 103] / V mg/l
Total Dissolved Solids
Weight of china dish after it has completely W5 mg
cooled with residue left after evaporating the
3
filtrate.
Total Dissolved Solids WD = W5 – W4 mg
Total Dissolved Solids (per ml of sample) [WD x 103] / V mg/l
Total Suspended Solids

Total Suspended Solids WS = WT – WD mg
= Total Solids – Total Dissolved Solids.
Total Suspended Solids (per ml of sample) [WS x 103] / V mg/l
Total Volatile Solids
Total Volatile Solids WV = WT – WF mg
= Total Solids – Total Fixed Solids.
Total Volatile Solids (per ml of sample) [WV x 103] / V mg/l
Total Settleable Solids
Settleable solids v ml
Settleable solids (per ml of sample) Wss = v/V ml/l
Result:
Conclusion:
4
Experiment No.: 02
ELECTRICAL CONDUCTIVITY
Aim
To determine the Electrical Conductivity of water.
Apparatus
Self-contained conductance instruments: (Conductivity meter).

Thermometer, capable of being read to the nearest 0.1°C and covering the range 10-50°C.
Conductivity Cells:
Reagents
1] Conductivity water
2] Standard potassium chloride solution [0.01M KCl]
Theory
Conductivity is a measure of the capacity of water to pass an electrical current. Conductivity

in water is affected by the presence of inorganic dissolved solids such as chloride, nitrate,
sulfate, and phosphate anions (ions that carry a negative charge), or sodium, magnesium,
calcium, iron, and aluminum cations (ions that carry a positive charge). A failing sewage
system would raise the conductivity because of the presence of
chloride, phosphate, and nitrate; an oil spill would lower conductivity. The basic unit of
measurement of conductivity is the mho or siemens.
Procedure
1. Calibrate the cell with solution.

2. Rinse the cell thoroughly with deionised distilled water and carefully wipe with a
tissue paper.
3. Dip the cell into the sample solution, swirl the solution and wait upto 1 minute for a
steady reading.
4. Note down the instrument reading and also temperature by a thermometer.
Description of sample Temperature Conductivity
Result:
Conclusion:
5
Experiment No.: 02
DETERMINATION OF CHLORIDES IN WATER
Aim
To determine the chloride in water by Mohr's method.

Apparatus
Burette, pipette, conical flask, measuring jar.
Reagents
1) 0.0141 N Silver nitrate (AgNO3)
2) Potassium chromate (K2CrO4) indicator
Theory
If water containing chlorides is titrated with silver nitrate solution, chlorides are precipitated
as white silver chloride. Potassium chromate is used as indicator, which supplies chromate
ions. As the concentration of chloride ions approaches extinction, silver ion concentration
increases to a level at which reddish brown precipitate of silver
chromate is formed indicating the end point.
NaCl + AgNO3 AgCl + NaNo3
2AgNO 3 + K2CrO4 AgCrO 4 + 2KNO3

(Reddish brown)
Procedure
1. Pipette out 50m1 of the given water sample into a clean conical flask.
2. Add 3 drops of Potassium chromate indicator. The solution turns yellow in colour.
3. Titrate this with 0.0141 N AgNO3 to a stable reddish brown or brick red precipitate.
Observations
Trial No Initial Reading Final reading Difference
Calculation
Chlorides = [A x N x 35.460 x 1000] / ml of sample = mg/l.

Where,
A = Volume of Silver nitrate consumed.
N = Normality of AgNO3 (0.0141)
Result:
Conclusion:
6
Experiment No.: 02
SULPHATE
Aim
To determine the concentration of sulphate in water by Turbidimetric method.
Apparatus
Nephelometer, Magnetic stirrer, Stopwatch, Measuring spoon 0.2 to 0.3 ml capacity, Nessler
tubes
Reagents
Barium chloride: crystals, 20-30mesh, Standard sulphate solution, Buffer solution A
and Buffer solution B: (required when the sample sulphate (SO4--) is less than 10mg/L).
Sample collection, preservation and storage

In presence of organic matter certain bacteria may reduce SO4-- to S--. To avoid this, store
sample at 4°C.
Principle
The turbidimetric method of measuring sulphate is based upon the fact that Sulphate ion is
precipitated in an acetic acid medium with Barium chloride (BaCl2) so as to form Barium
sulphate (BaSO4) crystals of uniform size and that this tendency is enhanced in presence of a
sodium chloride—hydrochloric acid solution containing glycerol and other organic
compounds. The absorbance of the barium sulphate solution is measured by a nephelometer
or turbidimeter and the sulphate iron concentration, determined by comparison of the reading
with a standard curve.
Procedure
1. Take suitable volume of sample and dilute to 100mL into a 250 ml Erlenmeyer flask.
2. Add 20 ml buffer solution, mix well by placing on a magnetic stirrer.
3. Keep the flask constantly stirred with the help of stirrer. Add 1-spatula BaCl2 crystals
with stirring and begin timing immediately.
4. Continue stirring at constant speed exactly for 1 minute after addition of BaCl2
5. After stirring pour some of the solution into the absorption cell of the photometer, and
measure the turbidity at 30 second intervals for four minutes.
6. Usually maximum turbidity occurs within two minutes and the reading remains
constant thereafter for 3 to 10 minutes. So, take reading with maximum turbidity
occurring in within four minutes.
7. Prepare a calibration curve. The standards are prepared at 5 mg/l increments in the 0–
40 mg/l sulphate range and their turbidity or absorbance read alternatively using a
spectrophotometer at 420nm with a light path of 2.5 to 10 cm.
8. Absorbance versus sulphate concentration is plotted and a curve is obtained.
9. Finding the absorbance for a given sample, the concentration of sulphates in the
solution is determined with the help of calibration curve.
7
Observations
Sample Size Turbidity/ Absorbance
0 [BLANK]
Unknown Sample
Calculation mg SO4--/l = {mg (SO4--) x 1000} / {ml of sample}
Result:
Conclusion:
8
Experiment No.: 03
ALKALINITY
Aim
To determine the alkalinity of water.

Apparatus
Burette, pipette, conical flask, measuring jar, volumetric flasks (1000ml, 200ml, 100ml)
Reagents
1. 0.02N H2SO4
2. Methyl orange indicator
3. Phenolphthalein indicator
Theory
Alkalinity of a sample is a measure of its capacity to neutralize acids. It is mainly due to the
salts of weak acids and some times due to weak or strong bases, mainly due to hydroxides,
carbonates and bicarbonates. Highly alkaline water leads to embitterment and causes
deposition of precipitates in boiler tubes. Bicarbonates of calcium and magnesium induce
temporary hardness to water. Other forms of alkalinity include Hydroxide alkalinity,
Carbonate alkalinity and Bicarbonate alkalinity. Generally high alkalinity waters are un-
potable and may physiologically affect the consumers. It is a very important parameter in
corrosion control.
Alkalinity due to
Value of P and T
OH CO3 HCO3
P=0 0 0 T
P<T/2 0 2P T-2P
P=T/2 0 2P 0
P>T/2 2P-T 2T-2P 0
P=T T 0 0
Procedure
Phenolphthalein Alkalinity
1. Take 25ml of sample (Water) in . a conical flask, add 2-3 drops of

phenolphthalein indicator.
2. Titrate the contents in the conical flask with 0.02N H 2 SO 4 . The end point
is pink to colorless.
9
3. Note the burette reading, which gives the volume of H 2 SO 4 Consumed.

Repeat till two concurrent values are obtained.
Observations
Calculation
P = [A × N × 50000] / ml of sample = mg/l as CaCO3
Where,
A = Volume of H 2 SO 4 consumed.
N = Normality of H2SO4 (0.02)
Result:
Procedure
Total Alkalinity
1. Take 25m1 of sample (Water) in a conical flask, add 2-3 drops of Methyl-orange
indicator.
2. Titrate the contents in the conical flask with 0.02N H2SO4. The end point is pale
yellow to pale pink.
3. Note the burette reading, which gives the volume of H2SO4.
4. Repeat until two concurrent values are obtained.
Observations
Calculation
T = [B ×N × 50000] / ml of sample = mg/l as CaCO3

Where,
B = Volume of H 2 SO 4 consumed.
N = Normality of H2SO4 (0.02)
10
Result:
Environmental significance:
1. Highly alkaline waters are usually unpalatable and consumers tend to seek other
supplies.
2. Chemically treated waters sometimes have rather high pH values, which have met with
some objection on the part of consumers.
3. Large amount of alkalinity imparts a bitter taste to water.
4. The principal objection of alkaline water is the reactions that can occur between
alkalinity and certain cations in waters. The resultant precipitate can foul pipes and
other appurtenances of water distribution systems.
Application of alkalinity in environmental practice:

1. Chemical coagulation of water and wastewater: To neutralize acids produced
during flocculation, the sample should be alkaline as otherwise further floc
formation slowly ceases.
2. Water is softening: To find out the quantity of lime and soda ash required for the
removal of hardness, alkalinity should be found out.
3. Corrosion Control: To control the corrosion due to acids, natural waters are
rendered to alkaline.
4. Effluents of wastewater: wastewaters containing excess hydroxide alkalinity are
not discharged into natural water bodies or sewers.
11
Experiment No.: 03
DETERMINATION OF ACIDITY IN WATER
Aim
To determine the mineral acidity and total acidity of the given water sample.
Apparatus
Burette, pipette, conical flask, measuring jar.
Reagents
1. 0.02 N Sodium hydroxide (0.02 N NaOH)

2. Methyl orange indicator
3. Phenolphthalein indicator
Theory
Acidity of a solution is a measure of its capacity to neutralize bases. Acidity is caused due to
the presence of mineral acids (Al2(S04)3, Fe2SO4), CO2, strong acids and weak bases. This is
caused mainly due to industrial wastes. Acidity of a sample is determined by titrating against
a strong solution of a base (alkali) such as NaOH. Types of Acidity include Mineral acidity
(partial acidity) and Total acidity.
Procedure
Mineral Acidity
1. Pipette out 25ml of the given water sample into a clean conical flask. Add 3
drops of methyl orange indicator to it. The solution becomes pink in colour.
2. Titrate this solution against 0.02 N NaOH solution taken in the burette till t he
colour changes from pink to pale yellow.
3. Not e do wn t h e b ur et t e r e adi n g w hi ch i n di c at e s t he v ol u m e of N a OH r un
dow n (consumed). Repeat the experiment until at least 2 concurrent readings are
obtained.
4. The End Point is Pink to pale yellow.
Observation
12
Calculation
P = [A × N × 50000] / ml of sample = mg/l as CaCO3
Where,
A = Volume of NaOH consumed.
N = Normality of NaOH (0.02)
Result:
Procedure
Total Acidity
1. Pipette out 25ml of the given water sample into a clean conical flask.
2. Add 3 drops of phenolphthalein indicator to it. The solution becomes colorless.
3. Titrate this solution against 0.02 N NaOH solution taken in the burette till the colour
changes to pink.
4. Note down the burette reading which indicates the volume of NaOH run down
(consumed).
5. The End Point is Colorless to pink.
6. Repeat the experiment until at least 2 concurrent readings are obtained.
Observations
Calculation
T = [B ×N × 50000] / ml of sample = mg/l as CaCO3

Where,
B = Volume of NaOH consumed.
N = Normality of NaOH (0.02)
Result:
13
Experiment No.: 03
pH
Aim
To determine the pH of the given water sample.

Apparatus
Burette, pH paper, pH meter, Glass Beaker, Buffer tablets.
Reagents
Standard Buffer solutions
Theory
pH is defined as the negative logarithm of hydrogen ion concentration or logarithm of
reciprocal of hydrogen ion concentration. Measurement of pH is one of the most important
and frequently needed tests in water and waste water engineering. Practically every phase of
water supply and wastewater treatment, for example acid base neutralization, water softening,
coagulation, disinfection and corrosion control is pH dependent. pH of drinking water should
be in the range of 6.5 to 8.5.
Procedure
1. By pH Paper Method
Tear off a piece of pH paper from the pH paper book. Dip this paper in a test
tube filled with the given water sample. Allow this paper to dry and develop
color. Compare the colour with the standard colour printed on the cover of the
pH paper book and record the pH. This method is an approximate method only.
2. By pH meter Method (Electrometric method)
1) Check the inside of the electrode of the pH Meter to ascertain whether it contains
sufficient level of saturated KO solution. The two electrodes are always immersed in
distilled water.
2) Set the temperature to 25°C (room temperature). Switch on the pH Meter and allow
the meter to warm up for about 15 minutes.
3) Take out the electrodes from the distilled water, rinse it and clean with a tissue paper
and immerse them in standard buffer solution of say pH 4. Press the knob to adjust it
to read pH position using the calibrating knob.
4) Repeat the procedure with the second buffer solution. If the meter shows the correct
reading directly, the instrument is calibrated
5) Place the electrodes in the given water sample and record the pH directly.
Result:
Conclusion:
14
Experiment No.: 04
HARDNESS TEST
Aim
To determine the Total, Permanent and temporary hardness of given water sample.
Apparatus
Burette, pipette, conical flask.
Reagents
1. 0.01M EDTA (Ethylene Di-amine Tetra Acetic Acid)

2. Ammonia buffer solution
3. Eriochrome black-T indicator.
Theory
Hardness in water is defined as that property which prevents lathering of soaps. It is caused
due to the presence of dissolved materials such as carbonates, bi-carbonates, chlorides,
nitrates and sulphates of calcium and magnesium.
Types of Hardness
1. Temporary hardness or Carbonate hardness

2. Permanent hardness or Non-carbonate hardness.
Temporary hardness in water is caused due to the presence of bicarbonates of calcium and
magnesium and boiling or addition of lime can easily remove this.
Permanent hardness is caused due to the presence of sulphates, chlorides and nitrates of
calcium and magnesium. This cannot be removed by boiling but requires special treatment
such as demineralization, ion exchange etc.
Hardness is expressed as mg/l on calcium carbonate scale. The desirable (permissible) limit
of total hardness in potable waters is 300mg/l and maximum permissible limit is 600 mg/l.
Principle
EDTA Titrimetric Method
A compound called Ethylene Di amine Tetra Acetic acid is made use of in this method. In
alkaline condition, the Ca2+ and Mg2+ develops a wine red colour with eriochrome black-T
indicator. When EDTA is added as a titrant, the Calcium and Magnesium ions get complexed
resulting in a sharp change from wine red to blue, which indicates the end point of the
reaction.
15
Procedure
Total Hardness
1. Pipette out 50m1 of the given water sample into a conical flask.
2. Add 1m1 of ammonia buffer solution and 3 drops of Eriochrome black-T indicator to
it. The solution turns wine red in color.
3. Titrate this solution against 0.01 M EDTA taken in a burette till the color changes
from wine red to blue which indicates the end point.
4. Note down this burette reading.
Observation
Permanent Hardness or Non Carbonate Hardness
1. Take about 60 ml of the given water sample, boil and cool it to remove temporary
hardness.
2. Transfer 50 ml of this solution into a clean conical flask.
3. Add l ml of ammonia buffer solution and 3 drops of Eriochrome black-T indicator to
it. The solution turns wine red in colour.
4. Titrate this solution against 0.01 M EDTA taken in a burette till the colour changes
from wine red to blue which indicates the end point.
5. Note down this burette reading which gives the volume of EDTA consumed.
Observation
16
Calculation
Total hardness = [A × B × 1000] / ml of sample = mg/l as CaCO3

Where,
A = Volume of EDTA consumed.
B = 1 (mg of CaCO3 equivalent to 1m1 of EDTA titrant)
Permanent hardness = [C × D × 1000] / ml of sample = mg/l as CaCO3

Where,
C = Volume of EDTA consumed for titrating the boiled and cooled sample.
D = 1 (mg of CaCO3 equivalent to 1m1 of EDTA titrant)
Temporary hardness = Total hardness - Permanent hardness
=…………-……….= ………… mg/l as CaCO3
Conclusion
17
Experiment No.: 04
HARDNESS TEST
Aim
To determine the Calcium-Magnesium hardness of given water sample.

Apparatus
Burette, pipette, conical flask.
Reagents
1. Std. EDTA (0.01 M)

2. 1 N NaOH solution
3. Muroxide indicator.
Significance
Small amounts of Ca combat corrosion of metal pipes by forming a protective coating where
as appreciable Ca content, On the other hand, forms scales in boilers, pipes, utensils etc.
Principle
When EDTA is added to water containing both Ca2+ and Mg2+, it combines with Ca2+ in
preference to Mg2+. When the pH is sufficiently high (12 to 13), Mg2+ is largely precipated as
hydroxide and Ca2+ can be determined directly using EDTA.
Procedure
1. Pipette out 100m1 of water sample into a conical flask.

2. Add 2m1 of 1 N NaOH, followed by 2 to 3 drops of Muroxide indicator and shake
well. The solution turns pink in colour.
3. Titrate this solution against Std EDTA solution taken in a burette and discontinue
the titration when the colour changes from pink to purple.
4. Note down the volume of EDTA run down from the burette.
18
Observation
Calculation
Calcium Hardness = [A × B × 1000] / ml of sample = mg/l as CaCO3
Where,
A = Volume of EDTA consumed.
B = 1 (mg of CaCO3 equivalent to 1ml of EDTA titrant)
Magnesium Hardness = Total hardness - Calcium hardness = mg/l as CaCO3

Concentration of Ca or mg/l of Ca = {A × B × 400.8} / ml of sample.
Concentration of Mg or mg/l of Mg = 0.243 × Magnesium hardness.
Result :
Conclusion:
19
Experiment No.: 05
DISSOLVED OXYGEN
Aim
To determine the Dissolved Oxygen content present in the given water sample.
Apparatus
BOD bottles, burette, pipette, conical flask, measuring jar.
Reagents
1. Manganous sulphate solution

2. Alkali Iodide azide
3. Concentrated Sulphuric acid
4. Starch solution
5. 0.1N Stock Sodium-thio-sulphate solution
6. 0.025 N Sodium-thio-sulphate (Na2S203)
Principle (The Winkler method with Azide modification)
When MnS04 and alkali iodide azide are added to water containing DO, the oxygen oxidizes
Mn2+ to give a precipitate of Mn(OH)2. By the addition of H2SO4, free iodine is liberated,
which is converted to blue colour by the addition of starch.
Procedure
1. Fill up the BOD bottle with the given water sample up to the neck.
2. Add 2ml of MnSO4 and 2ml of alkali iodide azide solution to the BOD bottle. Exclude
the air bubbles and mix the contents of the bottle by inverting the bottle a few times.
3. After the precipitate has settled to about 1/3rd the volume of the BOD bottle from the
bottom, add 2 ml of concentrated sulphuric acid. Once again mix the contents gently
till the suspension is completely dissolved and a uniform yellow colored solution is
obtained.
4. Take 203 ml of this solution in a conical flask and titrate with 0.025 N Na2S2O3 to a
pale straw color.
5. Add 1 ml of starch, which turns the solution blue in color. Continue the titration till
the blue color disappears. Note down the volume of Na2S203 consumed, which gives
the DO directly.
6. Hence the endpoint changes colour from Yellow to pale straw to blue to colorless.
Note
1 ml of 0.025 N Na2S203 is equivalent to 0.2 mg of oxygen.
20
Observation

1
2
3
DO in mg/L = ((0.2 x 1000) / 200) x volume consumed of thiosulphate of 0.025N
Result:
Conclusion:
21
Experiment No.: 05
BIO-CHEMICAL OXYGEN DEMAND
Aim
To determine the 5 day Bio-Chemical oxygen demand (BOD5) of the given sewage sample
with 2% dilute solution.
Apparatus
BOD bottles, burette, pipette, conical flask, measuring jar.
Reagents
1. Magnesium sulphate solution

2. Alkali Iodide azide
3. Concentrated Sulphuric acid
4. Starch solution
5. 0.025 N Sodium-thio-sulphate (Na2S203)
6. Phosphate buffer
7. Calcium chloride
8. Ferric chloride
Theory
BOD is the amount of oxygen required to stabilize the organic matter present in wastewater.
The degree of magnitude of BOD depends on the amount of organic matter present in the
wastewater and temperature during the experiment. The rate of oxygen demand or the process
of stabilization on the organic matter is very high or rapid in the initial stage i.e. 1-7 days.
This is mainly due to the oxidation of carbonaceous matter present in sewage. The oxidation
of the rest is done due to nitrification. The time required for complete oxidation is infinite,
but it is limited to 20 days. For practical purpose the period of oxidation is taken as 5 days.
Then 70%-80% of oxidation will be completed within first 5 days itself.
Principle
The standard test temperature is 20°C. This temperature is taken as the temperature of natural
water. There are 2 methods for determining the BOD.
1. Direct method: In this method BOD is directly calculated. This method is applicable
only for wastewater whose BOD value is < 7mg/l.
2. Dilution method: This test is made with various dilutions.
The following dilutions are suggested,

a. 0.1% to 7% - Strong trade waste
b. 1% to 5% - for raw or settled waste
22
c. 5% to 15% - for treated effluent

d. 25% to 100% - river water
Detailed Procedure
1. Place the desired volume of distilled water in a 5 litre flask (usually about 3 litres 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 sample 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 neutralised 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 requisite concentrations as per dilution method
8. Mix the contents in the measuring cylinder.
9. Add this solution into two B.O.D. bottles, one for incubation and the other for
determination of initial dissolved oxygen in the mixture.
10. Prepare in the same manner for other concentrations and for all the other samples.
11. Lastly fill the dilution water alone into two B.O.D. bottles. Keep one for incubation
and the other for determination of initial dissolved oxygen.
12. Place the set of bottles to be incubated in a B.O.D. incubator for 5 days at 20°C. Care
should be taken to maintain the water seal over the bottles throughout the period of
incubation.
13. Determine the initial dissolved oxygen contents in the other set of bottles and note
down the results.
14. Determine the dissolved oxygen content in the incubated bottles at the end of
5 days and note down the results.
15. Calculate the B.O.D. of the given sample.
Procedure
1. Prepare the required dilution of the sewage sample.

(For 1% dilution 1/100 × 300= 3 ml of sewage is taken in the BOD bottle and diluted
to 300 ml with aerated distilled water).
2. Determine the DO of this sample, which is the zeroth day DO or initial DO.
3. In another BOD bottle, prepare the sample of same dilution as before. Keep this
sample in a BOD incubator at 20°C for determining the DO after 5 days to get D2
(final DO or DO after 5 days.)
Calculations
BOD5= (D1-D2)/P mg/l

D1= Zero day or initial DO (before incubation)
D2 = DO after 5 days (final or DO after incubation)
P = % dilution (1% = 1/100 = 0.01)
23
Observations for D1
Observations for D2
Result:
Conclusions:
24
Experiment No.: 06
CHEMICAL OXYGEN DEMAND
Aim
To determine the COD of the given sewage sample by Open Reflux method.
Apparatus
Reflux apparatus of capacity 500m1 with a condenser, Burner or hot plate, 250 or 500mL
Erlenmeyer flask with standard (24/40) tapered glass joints, Friedrich’s reflux condenser (12
inch) with standard (24/40) tapered glass joints, Volumetric pipettes (10, 25, and 50ml
capacity), Burette, 50ml with 0.1ml accuracy, Burette stand and clamp, Analytical balance,
accuracy 0.001gm, Spatula, Volumetric flasks (1000ml capacity), glass beads.
Reagents
0.25N Std Potassium-dichromate (0.04167 M), Conc. Sulphuric acid reagent, 0.1 N Std. FAS,
Ferroin indicator and Mercuric sulphate crystals (analytical grade)
Principle
The organic matter gets oxidized completely by K2Cr2O7 in the presence of H2SO4 to produce
CO2 and H2O. The excess K2Cr2O7 remaining after the reaction is treated with Ferrous
Ammonium Sulphate .The dichromate consumed gives the oxygen required for oxidation of
the organic matter.
Procedure
Standardisation
Dilute 10 ml standard K2Cr2O7 to about 100 ml, add 30 ml conc.H2SO4, cool. Add 2 drops of
ferroin indicator and titrate with FAS.
Estimation
Place 0.4 gm HgSO4 in a reflux flask. Add 20 m1 sample or an aliquot of sample diluted to
20m1 with distilled water. Mix well and add glass beads followed by 10 m1 Std K2Cr2O7.
Add slowly 30 m1 H2SO4 containing Ag2SO4, mixing thoroughly. Connect the flask to the
condenser and mix the contents before heating. Reflux for a minimum period of 2 hours, cool
and wash down the condenser with distilled water. Dilute to about 150 m1, cool and titrate
excess K2Cr2O7 with 0.1 N FAS using Ferroin indicator. Sharp colour change from blue
green to wine red indicates the end point. Reflux the blank in the same manner using distilled
water instead of the sample.
25
Calculation
COD (mg/l) = [ ( a – b) × 8000 × N ] / ml of sample.

Where a & b = ml of FAS for blank and sample respectively.
N = Normality/Molarity of FAS
Observation
Result:
Conclusions:
26
Experiment No.: 07
AVAILABLE CHLORINE
Aim
To determine the Percentage of Chlorine in Bleaching Powder.
Apparatus
Conical flask, burette, pipette, measuring jar, funnel.
Reagents
Bleaching powder, Potassium Iodide (KI), Glacial Acetic acid, Starch solution and 0.1 N
Sodium thio sulphate
Theory
Chlorine a strong oxidizing agent and a very effective disinfectant used for the destruction of
pathogenic bacteria. Chlorine is generally applied in the form of bleaching powder for
disinfection of water. Commercial bleaching powders generally contain 25 to 30% of
available chlorine. This percentage is a very critical factor for effective disinfection of water.
Procedure
1. Weigh 5 gm of fresh bleaching powder and make it into a paste by adding a small
quantity of distilled water.
2. Add some more water, stir it and allow it to settle for a few minutes. Decant the
supernatant solution and dilute to 500 ml with distilled water. This is called chlorine
water or standard chlorine solution
3. Pipette out 25 ml of this chlorine water into a conical flask. Add 10 ml of KI and 10
ml of glacial acetic acid. Wait for 15-20 minutes (contact period)
4. Titrate this solution with 0.1 N Sodium thio sulphate. Add 2 drops of starch solution.
The solution becomes blue in color.
5. Continue the titration till the blue color just disappears. Note down the volume of
Sodium thio sulphate used.
Observation
27
Calculation
Percentage of chlorine in the given bleaching powder = [(X × 20) / 5] ×100
Where X = Amount of chlorine in bleaching powder = 0.003546 × titre value.
Where l ml of 0.1 N Na2S203 is equivalent to 1 milli equivalent of chlorine
i.e., 1 ml of 0.1 N Sodium thio sulphate = 0.003546 gm of chlorine
Note
5 ml of chlorine water is taken from 500 ml of the std. chlorine solution, hence 20 (500/25)
appears in the numerator of the above equation
Result:
Conclusions:
28
Experiment No.: 08
RESIDUAL CHLORINE
Aim
To determine the residual chlorine present in the given sample of water by Iodometric
method.
Apparatus
Conical flask, burette, pipette, measuring jar, funnel, beakers.
Reagents
0.001N Sodium-thio-sulphate, Glacial Acetic acid, KI solution & Starch solution indicator.
Principle
Chlorine present in water reacts with KI and iodine is liberated, when starch is used as
indicator, the presence of blue color produced indicates the presence of Iodine.
Procedure
1. Take 100m1 of sample in a conical flask.

2. Add 5ml of KI and 5ml of Glacial acetic acid and wait for about 15 minutes.
3. Titrate this solution against (0.001 N) Sodium thio sulphate until an endpoint
of a pale yellow color is obtained.
4. Add 2 to 3 drops of starch solution, when the solution becomes blue.
5. Continue the titration till the blue color turns colorless.
Chlorine in aqueous solution is not stable, and the chlorine content of samples or solutions,
particularly weak solutions, will decrease rapidly. Exposure to sunlight or other strong light
or agitation will accelerate the reduction of chlorine. Therefore, chlorine determination
should be done immediately after sampling, avoiding excessive light and agitation. The
sample should not be stored for analyses of residual chlorine.
29
Observation
Calculation
Amount of residual chlorine = [A x N x 35.46 x 1000] / ml of sample = mg/l.
Where
A = Volume of sodium thio sulphate run down.
N = Normality of sodium thio sulphate.
Result:
Conclusions:
30
Experiment No.: 08
CHLORINE DEMAND
Aim
To determine the chlorine demand of the given water sample.

Apparatus
BOD bottles, conical flask, burette, pipette, measuring jar, funnel, beakers.
Reagents
0.01N Sodium thio sulphate, KI solution, Glacial acetic acid and Starch solution
Theory
Chlorine is used as a disinfectant in water. The chlorine demand of water is the difference
between the amount of chlorine applied and the amount of free chlorine left after the contact
period as residue. The optimum dose of chlorine for given water is therefore generally
determined experimentally by adding varying amounts of chlorine to the given sample and
observing the residual chlorine left after a contact period of about 20 minutes. The dose,
which leaves a residual of 0.2 mg/l, is then selected. This total dose minus the free residual
will automatically represent the chlorine demand of water.
Procedure
1. Take 6 clean BOD bottles and add 200m1 of homogenous water sample to each.
2. Add 2, 4, 6, 8 ml etc of chlorinated water (known strength) to each bottle respectively.
3. Shake them and allow it to remain so for about 20 minutes. Chlorine reacts with the
organic matter and oxidizes it and also kills the bacteria present in the sample
4. After 20 minutes of contact period add 10 ml of KI and 10 ml glacial acetic acid to each of
these bottles.
5. Add 2 to 3 drops starch indicator. The appearance of blue color indicates the presence of
chlorine.
6. To estimate the amount of chlorine left in the sample as residue, it can be titrated against
0.01 N Na2S203 till the end point is reached, which is indicated by the disappearance of blue
color.
7. The chlorine demand is given by the difference between the applied Chlorine and the
residual chlorine after the contact period.
Observations for Residual Chlorine
31
Calculation
Sl.No BOD ml of chlorine mg of chlorine Residual Chlorine

bottle No added added to 1000 chlorine Demand [mg/l]
ml [mg/l]
Result:
Conclusions:
32
Experiment No.: 09
TURBIDITY
Aim
To determine the turbidity of the given sample by Nephelo turbiditymeter.
Apparatus
Nephelo turbiditymeter.
Reagents
Standard stock solution for Nephelo turbidity test
Determine turbidity on the day the sample is taken. It storage is unavoidable, store the
samples in dark for up to 24 hr. Prolonged storage before measurement is not recommended
because irreversible changes in turbidity may occur. All samples should be shaken vigorously
before examination.
Theory
Suspension of particles in water interfering with the passage of light is called turbidity.
Turbid water is undesirable from aesthetic point of view in drinking water supplies and may
also affect products in industries. It also causes a number of problems in water treatment
plants. The permissible limits for drinking water is 5 to 10 ppm, measured on a silica scale
Principle
When light is passed through a sample having suspended particles, some light is scattered by
the particles. The scattering of light is proportional to the turbidity.
Procedure
1. Switch on the Nephelo Meter and wait for 10 to 15 minutes for the instrument to warm
up.
2. Fill up the sample holder with distilled water, place it in the instrument & Close the
cover. Using the zero adjustment knob, adjust the reading to show zero.
3. Take out the distilled water sample and fill up the holder with the standard solution of
500 NTU from the already prepared standard stock solution. Place it in the instrument
and close the cover. Turn on the range switch to 1000 and adjust the instrument to read
50 (500 NTU) using the calibrating knob. Repeat the procedure with other standard
solutions to calibrate the instrument.
4. To know the turbidity of the given unknown sample, Keep it in the instrument and
record the turbidity reading directly in NTU.
33
Note:
Linear dilution of 1000 NTU (Stock solution)
Strength Dilution
1000 NTU Direct

800 NTU 80m1 of 1000 NTU (stock) solution + 20m1 of distilled water
Result:
Conclusions:
34
Experiment No.: 09
JAR TEST EXPERIMENT

Aim
To determine the optimum dosage of the coagulant alum to treat turbid water.
Apparatus
Glass beakers, Jar test apparatus, Nephelometer, Pipette, pH meter.
Reagents
Alum
Theory
The public water supplies consists of very fine particles of clay, silt etc which are present in a
finely divided state and it is not possible to detain them in primary sedimentation tanks unless
they are designed for longer detention periods. The other way to remove them is to increase
their size so that they become settleable, by adding certain chemicals called coagulants,
which react with impurities in water and convert them into settleable size. Some common
coagulants used are Alum, Sodium aluminate, ferric chloride, copperas, chlorinated copperas,
ferric sulphate etc.
Procedure
1. Prepare standard solution of coagulant say 1% solution, by dissolving 5gm of alum in

distilled water and making upto 500m1.
The strength of the solution = 1% (1m1 = 10mg)
2. Take 1-litre beakers and fill them with sample up to the mark.
3. Keep each beaker below each paddle and lower the paddles, such that each one is
about 1cm above the bottom.
4. Find the pH of the sample and adjust it to 6 to 8.5.
5. Pipette 1, 2, 3, 4, 5, 6 ml of the alum solution into the test samples, i.e. the dosage is
10, 20, 30, 40, 50 and 60mg respectively.
6. Immediately run the paddles at 100 rpm for 1 minute.
7. Reduce the speed to 30–40 rpm and run at this rate for 30 minutes.
8. Stop the machine, lift out the paddles and allow to settle for 30 minutes.
9. Find the residual turbidity of the supernatant using nephelometer.
10. Plot a graph with alum dosage along x-axis and turbidity along y-axis.
11. The dosage of alum, which represents least turbidity, gives Optimum Coagulant
Dosage.
12. Repeat steps 1–10 with higher dose of alum, if necessary.
Note
Strength of solution. 1 gm = 100m1
35
Observation
Jar ml of mg of Concentration
Description of
No coagulant coagulant mg/l
the sample added added
Result:
Conclusions:
36
Experiment No.: 10
IRON
Aim
To determine the concentration of iron in the given water sample by Phenanthroline method.
Apparatus
Nessler's tubes (100 ml), Conical flasks, Pipettes, Colorimeter.
Reagents
Conc. HCl acid, Hydroxylamine hydrochloride solution, Ammonium acetate buffer solution,
1, 10-Phenanthroline solution, Stock Fe soln. and Std Fe soln. (1ml = 1 mg Fe).
Principle
In the Phenanthroline method, the ferric form of iron is reduced to ferrous form by boiling
with hydrochloric acid and hydroxylamine hydrochloride. The reduced iron chelates with
1,10 phenanthroline at pH 3.2 - 33 to form a complex of orange-red color. The intensity of
color is proportional to the concentration of iron and obeys Beer's law.
Procedure
1. Pipette 10, 20, 30 and 50m1 of standard iron solution into 100m1 conical flask
2. Add l ml of Hydroxyl amine hydrochloride solution and 1m1 of sodium acetate
solution to each flask.
3. Dilute each to about 75m1 with distilled water and Add 2ml of 1,10-phenanthroline
solution to each flask.
4. Make up the contents of each flask exactly to 100m1 by adding distilled water and
allow atleast 10-15 minutes for maximum color development.
5. For visual comparison pour the solution in 1.00 m1 Nessler tubes and keep them in a
stand. Mix the sample thoroughly and measure 50ml into a conical flask
8. Add 2 ml of concentrated HCl and 1 ml of Folderol amine solution and a few glass
beads. Boil the contents to half of the volume for dissolution of all the iron
9. Cool the flask to room temperature and transfer the solution to a 100 m1 Nessler tube
10. Add 10m1 of Ammonium acetate buffer solution and 2m1 Phenanthroline solution
and dilute to 100m1 mark with distilled water. If the sample contains interference of
heavy metals add 10m1 of Phenanthroline instead of 2m1
11. Mix thoroughly and allow atleast 10-15 minutes for maximum color development
12. For visual comparison, match the color of sample with that of the standards prepared
in steps 1-6 above
13. The matching color standard will give the concentration of iron in the sample
For photometric measurement, use light path at 510nm. Read standards against distilled water
set at zero absorbance and plot a calibration curve, including a blank. If samples are coloured
37
or turbid, carry a second set of samples through all steps of the procedure without adding
Phenanthroline.
Comments
Environmental Significance
Long time consumption of drinking water with a high concentration of iron can lead to
liver diseases. Iron rich water exposed to the air becomes turbid and highly unacceptable
from the aesthetic view point.
38
Experiment No.: 11
FLUORIDE
Aim
To determine the concentration of fluoride for a given water sample by Colorimetric method
(SPANDS method).
Apparatus
Distillation apparatus, Colorimetric, Nessler tubes [capacity-100ml], Pipette
Reagents
1. Standard sodium fluoride solution

2. Acid zirconium alizarin reagent
Polyethylene bottles are preferred for collecting and storing samples for fluoride analysis.
Glass bottles are satisfactory, provided that they have not previously contained high-fluoride
solutions. Always rinse the bottle with a portion of the sample.
Principle
This method is used for estimation of fluoride in natural water in the concentration range 0-
1.4 mg/l. The test is based on the fact that fluoride ion combines with zirconium on to form a
stable complex ion, ZrF6 and this results in bleaching the reddish colour of Zirconium and
alizarin combination. The decrease in intensity of colour is directly proportional to fluoride
concentration.
Procedure
1. The sample should be free from chlorine, if chlorine present, it shall be dechlorinated
with a slight excess of sodium thio sulphate solution before use.
2. If the sample contains excess interfering ions, the sample should be appropriately
diluted and distillated before test.
3. Take 0, 1, 2, 6, 8, 14 ml of Standard sodium fluoride solution in six Nessler tubes.
[The range should be such that it is between 0 and 1.4 mg/l]
4. Add 5m1 of acid zirconium reagent in each Nessler tube and make it up with distilled
water.
5. Similarly add 5m1 of acid zirconium reagent into the Nessler tubes containing sample
and make it up with distilled water.
6. Mix thoroughly and compare the colors after standing for one hour.
39
Comments
Fluoride ions have dual significant in water supplies. High concentration of F - causes dental
fluorosis (disfigurement of the teeth) and skeletal fluorosis. At the same time, a concentration
less than 0.8mg/l results in ‘dental caries’. Hence, it is essential to maintain the F-
concentration between 0.8 to 1.0mg/l in drinking water.
40
Experiment No.: 12
MPN TEST{B/E-COLI) COLIFORMS INDEX}
Aim
To determine the most probable number (MPN) index of coliforms and E-coli organisms in
the given water sample by multiple tube fermentation tube technique.
Apparatus
Incubator, fermentation tubes, test tubes, EMB agar plate, Petri-dishes, microscopes,
Autoclave, Pipettes, Measuring jars, Inoculating equipments [platinum loop], Media
preparation utensils.
Reagents
1. Lauryl tryptose broth

2. Brilliant green lactose bile broth
3. Endo methylene blue agar.
4. Eosin methylene blue agar.
5. Lactose broth
Theory
Coliform group comprises of all the aerobic, facultative and anaerobic gram-negative non-
spore forming rod shaped bacteria that ferment lactose with gas formation within 48 hours at
35°C. The standard test for this group may be carried out either by multiple tube fermentation
technique or by membrane filter technique. Escherichia coli (E.coli) is used as an indicator
organism for the purpose of sanitary examination of water. The E.coli test by multiple tube
fermentation technique consists of 3 phases – presumptive, confirmed and completed. The
results are expressed in terms of MPN (Most Probable Number), which is based on certain
probability formulae. The estimate may give a value greater than the actual number of
coliform present. The accuracy of any single test depends on the number of tubes fermented.
This method helps in describing the sanitary quality of water. The safety of the water is
generally judged from the knowledge of sanitary condition and mentioned by the number of
samples yielding positive or negative results. If more than 95% should yield negative results,
the safety is usually assured.
Procedure
1. Collect the sample in sterilized bottles

2. Prepare the sterilized media necessary for the bacteriological test and keep them re0y
in test tubes containing Durham tubes / fermentation tubes.
3. Inoculate the sample in an exponential order le 10, 1 and 0.1 ml in 5 tubes each of
lauryl tryptose broth under asceptic conditions.
4. Incubate all the tubes at 35°C.
41
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. Remaining tubes are further incubated at 35°C for another 24 hours.
This is the presumptive test for coliform organisms.
7. One or two loop full of the liquid from the positive lauryl tryptose tubes are transferred
into the sterilized EC medium tubes and incubated for 24 hours at 44°C.
8. The gas production after 24 hours confirms the presence of E-Coli or feacal Coliforms.
42
43
Observations
Computation of MPN
The number of positive finding of coliform group organisms resulting from the multiple
portion decimal dilution planting should be computed as the combination of positives and
recorded in terms of the Most Probable Number (MPN). The MPN may be estimated by
Thomas’ simple formula:
Result : MPN/100 ml =
Conclusion:
44
Experiment No.: 13
NITRATES
Aim
To determine the concentration of Nitrate Nitrogen present in the given sample of given
water sample.
Apparatus
Spectrophotometer or Colorimeter, Nessler tubes (capacity-100ml), Beakers (capacity-100ml

and Water bath.
Reagents
1. Stock nitrate solution

2. Standard nitrate solution
3. Standard silver sulphate
4. Phenol di sulphonic acid (PDA)
5. Concentrated Ammonium hydroxide [NH4OH].
Principle
Nitrate reacts with Phenol di sulphonic acid and produces a nitro-derivative which in alkaline
medium develops a yellow color. The color produced follows the Beer's law and is directly
proportional to the concentration of nitrate present in the sample.
Procedure
1. Take 50ml of filtered sample in an Erlenmeyer flask.

2. Add an equivalent amount of silver sulphate to remove chlorides
(1 mg/l Cl = l ml Ag2SO4 solution)
3. Heat slightly and filter the precipitate of AgCl.
4. Evaporate the filtrate in a porcelain disc to dryness.
5. Cool and dissolve the residue in 2m1 phenol di sulphonic acid and dilute to 50 ml.
6. Add 10 m1 of liquid ammonia to develop the yellow color.
7. Read the color developed at 410 nm with a light path of 1cm.
8. Calculate the concentration of nitrate nitrogen from the standard curve.
9. Prepare the standard curve using suitable aliquots of standard nitrate solution in the
range of 5-500 mg NO3-N following the above procedure.
Interferences
Chlorides and nitrite are the two main sources of interferences. Pretreatment of sample is
necessary when the interfering radicals are present.
45
Result:
Conclusion:
In surface waters, nitrate may cause objectionable eutrophication. Nitrates are poisonous to
infants, animals including humans. In humans it can cause 'methemoglobinemia'.
46
Experiment No.: 14
SODIUM
Aim
To determine the concentration of Sodium in given water sample by Flame Photometer.
Apparatus
Flame photometer
Reagents
Stock sodium solution, Intermediate sodium solution, Standard sodium solution
Storage
Store samples containing low sodium concentrations or alkaline samples in polyethylene
bottles to eliminate the possibility of sample contamination due to leaching of the glass
container.
Principle of Quantification
Trace amounts of sodium can be determined by flame emission photometry at the wavelength
of 589 nm. The sample is sprayed into a gas flame and excitation is carried out under
carefully controlled and reproducible conditions. The calibration curve may be linear but has
a tendency to levels off at higher concentrations.
Procedure
1. Follow instructions of flame photometer manufacturer for selecting proper photocell,

wavelength, slit width adjustments, fuel gas and air pressure, steps for warm up,
correcting for interference and flame background, rinsing of burner, sample ignition
and emission intensity measurements.
2. Prepare a blank and sodium calibration standards, in any of the applicable ranges, 0-
100, 0-10, or 0-1 mg Na/l. Set instrument zero with standard containing no sodium.
3. Measure emission at 589 nm and prepare calibration curve.
4. Determine sodium concentration of the sample, or diluted sample, from the curve..
Calculation
Dilution mg Na/l = mg Na/l [from the calibration curve] x Dilution
Where,
47
Observations
Sample Size Absorbance

0
5
10
15
25
Unknown Sample
Result:
Conclusions:
48
Experiment No.: 14
POTASSIUM
Aim
To determine the concentration of Potassium in given water sample by Flame Photometer.

Apparatus
Flame photometer
Reagents
Stock Potassium solution, Intermediate potassium solution, Standard potassium solution
Storage
Do not store samples in soft-glass bottles because of the possibility of contamination from
leaching of the glass. Use acid washed polyethylene or borosilicate glass bottles. Adjust
sample to pH <2 with nitric acid. This will dissolve potassium salts and reduce adsorption on
vessel walls.
Principle of Quantification
Trace amounts of potassium can be determined in either a direct-reading of internal standard
type of flame photometer at a wavelength of 766.5 nm. The sample is sprayed into a gas
flame and excitation is carried out under carefully controlled and reproducible conditions.
The calibration curve may be linear but has a tendency to levels off at higher concentrations.
Procedure
1. Follow instructions of flame photometer manufacturer for selecting proper photocell
wavelength, slit width adjustments, fuel gas and air pressure, steps for warm up,
correcting for interference and flame background, rinsing of burner, sample ignition
and emission intensity measurements.
2. Prepare a blank and potassium calibration standards, in any of the applicable ranges, 0-
100, 0-10, or 0-1 mg/l. Calibrate instrument, set instrument to zero absorbance with
blank solution containing no potassium.
3. Measure emission at 766.5 nm and prepare calibration curve.
4. Determine potassium concentration of the sample, or diluted sample, from the curve.
Calculation
mg K/l = mg K/l [from the calibration curve] x Dilution
Where,
49
Observations
Sample Size (ml) Absorbance
BLANK
Unknown Sample
Result:
Conclusions:
50
Environmental Lab viva questions

1. Define the term solids? Mention various types of solids present in
sewage sample.
2. What is the significance of determining settleable solids?
3. Distinguish between fixed solids and volatile solids? What is the
allowable limit of total solids in drinking water?
4. Mention the methods used to determine the sulphate?
5. Mention the possible source of chloride entry into the sewage?
6. What are the causes for the acidity and alkalinity of water?
7. How the acidity of the water/wastewater can be eliminated?
8. Why acidity, alkalinity and hardness are expressed in terms of caco3?
9. Mention the principle involved in pH meter?
10.Mention various methods used to measure pH of a water/waste water
sample?
11.Mention the sanitary significance of pH of water /waste water?
12.Define hardness of water? Mention the types of hardness?
13. Mention the causes of hardness of water?
14.How do you find temporary and permanent hardness of water sample?
15.Mention the various bacteriological tests conducted on water sample?
16.What is E-coli test? What is the sanitary significance of conducting E-coli
test?
17.Why it is necessary to maintain the minimum DO in water bodies? What
is its amount?
18.Why in DO test 201/203 ml of sample is taken instead of 200ml sample?
19.Mention the various forms in which chlorine is applied to water sample?
20.Distinguish between disinfection and sterilization?
21. What do you mean by break point chlorination?
22.What is chlorine demand?
23.Mention the various types of chlorination?
24.Why do you determine residual chlorine in water treatment 0practice
and water supply mains?
25.Free available chlorine is more effective than combined available
chlorine .why?
26.What is coagulant? Why alum is used as universal coagulant?
27.Mention the factors affecting the dosage of coagulant?
51
28.Mention the causes of turbidity in water? How turbidity is measured?

29.What is the principle involved in turbidity meter?
30.What is the principle involved in flame photometer?
31.What is the principle involved in spectrophotometer?
32.What for jar test apparatus is used?
33.What is the difference between BOD and COD?
34.Differentiate between E-coli and B-coli?
35.What is the effect of iron if it is in excess concentration in drinking
water?
36.How iron is removed in water?
37.What are the adverse effect of nitrates on human health and
environment if its concentration exceeds the limit in water?
38. How nitrate concentration from water bodies can be reduced?
39.Name few causes for taste and odour in water supplies
40. What are the ill-effects of fluorides when they are not in the permissible
limits?
41.Why bacteriological test should be necessary in handling problems of
water supply?
42.What are water-borne diseases and how it can be prevented?
43. What is the percentage of chlorine in fresh bleaching powder?
44. What are the methods of deflouridation ?
45. What is coagulation? What are the common coagulants used in water
treatment plant?
52
TABLE ISI Tolerance Limits for the Sewage and Industrial Effluents and that of Inland Surface
Water.
Characteristics Tolerance limits Tolerance limits for industrial effluents Tolerance limits for
for sewage discharged into Inland surface water,
effluents Inland surface water Public when used as raw water
discharged into sewers for public water supplies
inland surface IS: 2490-1974 IS: 3306-1974 and bathing ghats
water IS: 4764- IS: 2296-1974
1973
1 2 3 4 5
BOD ( 5day 20 30 500 3
200C)
mg/l
COD, mg/l - 250 - -
pH - 5.5-9.0 5.5-9.0 6.0-9.0
Total suspended 30 100 600 -
solids mg/l
Temperature , 0C - 40 45 -
Oil & Grease, - 10 100 0.1
mg/l
Phenolic - 1.0 5 0.005
compounds mg/l
Cyanides ( as - 0.2 2.0 0.01
CN), mg/l
Sulphides ( as S), - 2.0 - -
mg/l
Fluorides (as F ), - 2.0 - 1.5
mg/l
Total residual - 1.0 - -
chlorine mg/l
Insecticides, mg/l - zero - zero
Arsenic (as AS) - 0.2 - 0.2
mg/l
Cadmium (as Cd) - 2.0 - -
mg/l
Chromium, - 0.1 2.0 0.05(Total chromium)
hexavalent
Copper, mg/l - 3.0 3.0 -
Lead, mg/l - 0.1 1.0 0.1
Mercury, mg/l - 0.01 - -
Nickel, mg/l - 3.0 2 -
Selenium, mg/l - 0.05 - 0.05
Zinc, mg/l - 5.0 15.0 -
Chloride,(as Cl), - - 600 600
mg/l
Sulphates, mg/l - - - 1000
% Sodium - - 60 -
Ammoniacal - 50 50 -
Nitrogen, mg/l
Nitrates(as No3) - - - 50
53
Radioactive
material
α- emitters , 10-7 - 10-9

µc/ml-
β-emitters, 10-6 - 10-8
µc/ml-
Dissolved - - - 40% of the saturation
Oxygen, mg/l value, or 3 mg/l,
Coliform whichever is higher.
organism – - - - Should not exceed
(monthly 5000.(should not exceed
average) 20000 with less than 5%
MPN per 100ml samples, and 5000 with
less than 20% samples)
54

Env Lab Manual 2020

Uploaded by

Document Information

Original Description:

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Env Lab Manual 2020

Uploaded by

Copyright:

Available Formats

ENVIRONMENTAL ENGINEERING LAB MANUAL CVL77

2. Discussion after each experiment should be based on the following points:

(a) Limit prescribed for that constituent in drinking water standards.

DOS AND DONOT’S IN THE LABORATORY

1. Do thoroughly clean the glassware before and after use.

Experiment Content Hours COs

Name & Signature of Faculty In-charge Head of the Department

Concurrent with the evaluation of environmental engineering practice, analytical

The objective of sampling is to collect representative sample. Representative

Complete and unequivocal preservation of samples, whether domestic

STANDARDS FOR DRINKING WATER

SOLIDS IN WASTE WATER

Determination of Solids in sewage: Total Solids, Suspended Solids, Dissolved Solids,

Classification of Sewage Solids:

Total solids in wastewater exist in 3 forms:

Dissolved solids, Colloidal solids and Suspended solids.

These are further classified into:

b] Total Fixed Solids

c] Total Dissolved Solids

W5 = Weight of empty evaporating dish in mg + Residue left after evaporating the

d] Total Suspended Solids = Ws = Total Solids – Total Dissolved Solids.

e] Total Volatile Solids = WV = Total Solids – Total Fixed Solids.

f] Settleable Solids (by volume) [WSS]

Observations, Calculations and Results

Parameter Label Unit Result

Total Fixed Solids

Total Suspended Solids

Settleable solids (per ml of sample) Wss = v/V ml/l

To determine the Electrical Conductivity of water.

Self-contained conductance instruments: (Conductivity meter).

Conductivity is a measure of the capacity of water to pass an electrical current. Conductivity

1. Calibrate the cell with solution.

Description of sample Temperature Conductivity

DETERMINATION OF CHLORIDES IN WATER

To determine the chloride in water by Mohr's method.

2AgNO 3 + K2CrO4 AgCrO 4 + 2KNO3

Trial No Initial Reading Final reading Difference

Chlorides = [A x N x 35.460 x 1000] / ml of sample = mg/l.

Sample collection, preservation and storage

Calculation mg SO4--/l = {mg (SO4--) x 1000} / {ml of sample}

To determine the alkalinity of water.

1. Take 25ml of sample (Water) in . a conical flask, add 2-3 drops of

3. Note the burette reading, which gives the volume of H 2 SO 4 Consumed.

Trial No Initial Reading Final reading Difference

Trial No Initial Reading Final reading Difference

T = [B ×N × 50000] / ml of sample = mg/l as CaCO3

Application of alkalinity in environmental practice:

DETERMINATION OF ACIDITY IN WATER

1. 0.02 N Sodium hydroxide (0.02 N NaOH)

Trial No Initial Reading Final reading Difference

2. Add 3 drops of phenolphthalein indicator to it. The solution becomes colorless.

5. The End Point is Colorless to pink.

6. Repeat the experiment until at least 2 concurrent readings are obtained.

Trial No Initial Reading Final reading Difference

T = [B ×N × 50000] / ml of sample = mg/l as CaCO3

To determine the pH of the given water sample.

Standard Buffer solutions

2. By pH meter Method (Electrometric method)

1. 0.01M EDTA (Ethylene Di-amine Tetra Acetic Acid)

1. Temporary hardness or Carbonate hardness

EDTA Titrimetric Method

Trial No Initial Reading Final reading Difference

Permanent Hardness or Non Carbonate Hardness