EE Lab Manual Final Draft

LABORATORY MANUAL
FOR
FNVIRONMFNTAL FNGINFFRING
GENERAL
I. Instructions
1. This laboratory manual is a reference manual for using the environmental science/engineering
laboratory.
2. Discussion after each experiment should be based on the following points:
(i) Limit prescribed for that constituent in drinking water standards.
(ii) The suitability of the sample for drinking purpose with respect to that particular constituent.
3. Users may refer the following for writing the discussion after each experiment:
(i) Standard Methods for the Examination of Water and Waste Water, American Public Health
Association, 1015, 15
th
Street, N.W., Washington D.C., 2005.
(ii) Chemistry for Environmental Engineers, Sawyer and McCarty, Tata Mc-Graw Hill.
(iii) Manual of Standards of Quality for Drinking Water Supplies, Indian Council of Medical
Research, New Delhi.
(iv) International Standards for Drinking Water World Health Organisation.
(v) IS 2490 - 1981, IS 3306 - 1974, IS 3307 - 1977, IS 7968 - 1976, IS 2296 - 1974, Bureau
of Indian Standards, New Delhi.
II. DOs and DONTs 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.
6. Do use large volumes of water, when a person is splashed with acid to prevent serious burns.
7. Do weigh the articles in a balance only at room temperature.
8. Do use different pipette for different reagents.
9. Do not pipette out acids and other toxic reagents by mouth.
10. 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.
III. General Information
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.
Normality
The desired normality of the titrant is obtained by the relationship of 1 to the equivalent weight of the measured
material. Thus normality of acid solution to measure ammonia, ammonia nitrogen, and alkalinity as CaCO
3
Ammonia
1/eq. wt. = 1/17 = N/17 = 0.0588N
Ammonia N 1/eq. wt. = 1/14 = N/14 = 0.0715N
Alkalinity 1/eq. wt. = 1/50 = N/50 = 0.020N
The normality of basic solution to measure mineral acidity as CaCO
3
is:
Acidity 1/eq. wt. = 1/50 = N/50 = 0.020N
The normality of silver nitrate to measure chloride and sodium chloride is:
Chloride 1/eq. wt. = 1/35.45 = N/35.45 = 0.0282N
Sodium chloride 1/eq. wt. = 1/58.44 = N/58.44 = 0.071N
Thus the substance measured is calculated as follows:
=
mL of titrant used 1, 000
mg / L
mLof sample
Most materials subjected to the analysis of water and wastewater fall in the realm of dilute solutions i.e., a few mg
in a litre. So the results are normally expressed in mg/L or ppm. Parts per million (ppm) is a weight ratio; but mg/L
is a weight by volume ratio. The relationship is given as follows:
ppm =
mg / L
Sp.gr.
If concentrations are less than 0.1 mg /L, express them in g/L (micrograms per litre).
If concentrations are more than 10,000 mg/L, they are expressed in percentages.
Plotting of Graphs
Rules listed by Worthing and Geffner are to be followed while plotting graphs. They are:
1. The independent and dependent variables should be plotted on abscissa and ordinate respectively.
2. The scale should be so chosen that the value of either coordinate could be found quickly and easily.
3. The curve should cover as much of the graph sheet as possible.
4. The scales should be so chosen that the slope of the curve approach unity as nearly as possible.
5. The variables should be chosen to give a plot that will be as nearly a straight line as possible.
2. Chemical substances
which may affect health
1. Toxic substances
Classification of Procedures
Laboratory analytical procedures are classified to quantify the chemical substances as follows:
1. Toxic chemical substances: e.g., lead, arsenic, selenium, hexavalent chromium, cyanide.
2. Chemical substances affecting health: e.g., fluoride, nitrate.
3. Chemical substances affecting potability: e.g., residue, turbidity, colour, taste and odour, iron, manganese,
copper, zinc, calcium, magnesium, sulphate, chloride, pH and phenolic compounds.
4. Chemical substances indicative of pollution: e.g., total organic matter, BOD, Kjeldahl nitrogen (total
organic nitrogen), albuminoid nitrogen, nitrite nitrogen and phosphate.
5. Residual chlorine.
Standards of Water Quality
Standards of water quality are presented as follows:
Bacteriological Quality
1. Treated water: In 90% of the samples examined throughout the year, the coliform bacteria shall not
be detected or the MPN index shall be less than 10. None of the samples shall have an MPN index
of coliform bacteria in excess of 10. An MPN index of 810 shall not occur in consecutive samples.
2. Untreated water: In 90% of the samples examined throughout the year, the MPN index of coliform
organisms should not be less than 10. None of the samples should show an MPN index greater than
20. An MPN index of 15 or more should not be permitted in consecutive samples.
Chemical and Physical Quality
Classification Substances Maximum allowable
concentration
Lead (Pb) 0.1 mg/L
Selenium (Se) 0.05 mg/L
Arsenic (As) 0.2 mg/L
Chromium (Cr
6+
) 0.05 mg/L
Cyanide (CN) 0.01 mg/L
Fluoride (F
) 0.081.0 mg/L
Nitrate
3
(NO ) 50.0 mg/L
Total solids 500 mg/L
Colour 5 Units
Turbidity 5 Units
Taste Unobjectionable
Odour Unobjectionable
Manganese (Mn) 0.1 mg/L
Contd...
Iron (Fe) 0.3 mg/L
Copper (Cu) 1.0 mg/L
Zinc (Zn) 5.0 mg/L
Calcium (Ca) 75 mg/L
Magnesium (Mg) 50 mg/L
Sulphate 200 mg/L
Chloride (Cl
) 200 mg/L
pH range 7.08.5
Phenolic substances 0.001mg/L
Significance and Determination of Chemical Parameters
Chemical parameters and their significance are presented as follows. The methods of the analysis adopted are also
presented. However, only simple methods will be dealt within this manual.
Chemical parameters commonly determined in natural waters and water supplies
No. Chemical species Significance in water Methods of analysis
commonly used
1. Acidity Indicative of industrial Titration
pollution, acid mine drainage
2. Alkalinity Water treatment, buffering, Titration
algal productivity
3. Ammonia Productivity, pollution Colorimetry
4. Calcium Hardness, productivity Atomic absorption
treatment
5. Carbon dioxide Bacterial action, corrosion Titration, calculation
6. Chloride Saline water contamination Titration, potentiometry
7. Chlorine Water treatment Colorimetry, titration
8. Fluoride Water treatment, toxic at Colorimetry, potentiometry
high level
9. Hardness Water quality, treatment Titration, atomic absorption
10. Iron Water quality, treatment Colorimetry, atomic
absorption
11. Magnesium Hardness Atomic absorption
12. Manganese Water quality Atomic absorption
13. Nitrate Productivity, toxicity Colorimetry, potentiometry
14. Nitrite Toxic, pollutant Colorimetry
3. Chemical substances
affecting the potability of
water
Contd...
15. Nitrogen (albumin.) Proteinaceous material Colorimetry
16. Nitrogen (organic) Organic pollution Colorimetry
17. Oxygen Water quality Titration, electrochemical
18. BOD Water quality, pollution Microbiological titration
19. COD Water quality, pollution Chemical oxidation-
reduction
20. pH Water quality, pollution Potentiometry
21. Phosphate Water quality, pollution Colorimetry
22. Sulphate Water quality, pollution Gravimetry, turbidimetry
23. Sulphide Water quality, pollution Colorimetry, potentiometric
titration
IV. Introduction
Humanity and Environment
A characteristic, which has set Homo sapiens apart from other species, has been their ability to control many
aspects of their environment. Throughout recorded history people have continually struggled to manage their
natural environment in order to improve their health and well-being. In recent years environmental sanitation in
many parts of the world has led to large reduction or virtual elimination of diseases spread via the environment.
Continuous environmental vigilance is necessary to keep away these weeds from the garden of humanity from
increasing out of proportion among a large part of the earths population.
Peoples success in the control of environmental borne diseases has not reduced the need for ever-increased
efforts of effective management of the total environment. The population explosion, an affluent society with desires
for a vast array of products, increased radiations, greater energy use, increased food production needs, and other
developments have created strains on parts of the ecological systems. Perhaps never in history have people
demonstrated such great concern for their total environment as is now being witnessed in many parts of the earth,
particularly in those areas which have benefited most from peoples environmental control efforts toward effective
use of human, material and natural resources. Over the years, intensification of old problems and the introduction
of new ones have led to basic changes in the philosophy of environmental engineering practice.
Water is one of the materials required to sustain life and has long been suspected of being the source of many
of the illnesses of man. It was not until a little over 100 years ago that definite proof of disease transmission through
water was established. Originally the major objectives were to produce hygienically safe water supplies and to
dispose off wastes in a manner that would prevent the development of nuisance conditions. Many other factors
concerned with aesthetics, economics, recreation and other elements of better living are important considerations
and have become part of the responsibilities of the modern environmental engineer.
The public has been more exacting in their demands as time has passed, and today water engineers are
expected to produce finished waters that are free of colour, turbidity, taste, odour and harmful metal ions. In
addition, the public desires water, which is low in hardness and total solids, non-corrosive, and non-scale forming.
To meet with such stringent standards, chemists, biologists and engineers must combine their efforts and talents
together and hence the need for analytical testing of water and waste becomes necessary.
Importance of Quantitative Analysis
Quantitative analysis serves as the keystone of engineering practice. 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 used by biologists.
Every problem in environmental engineering must be approached initially in a manner that will define the problem.
This approach necessitates the use of analytical methods and procedures in the field and laboratory, which have
proved to yield reliable results. Once the problem has been defined quantitatively, the engineer is usually in a
position to design facilities that will provide a satisfactory solution.
After construction of the facilities has been completed and they have been placed in operation, usually constant
supervision employing quantitative procedure is required to maintain economical and satisfactory performance.
The increase in population density and new developments in industrial technology are constantly intensifying old
problems and creating new ones. In addition, engineers are forever seeking more economical methods of solving
old problems. Research is continuously under way to find answers to the new problems and better answers to old
ones. Quantitative analysis will continue to serve as the basis for such studies.
Character of Problems
Most problems in environmental engineering practice involve relationships between living organisms and their
environment. Because of this, the analytical procedures needed to obtain quantitative information are in often a
strange mixture of chemical and biochemical methods, and interpretation of the data is usually related to the effect
on microorganisms or human beings. Also, many of the determination used fall into the realm of microanalysis
because of the small amounts of contaminants present in the samples. Ordinarily, the amounts determined are a few
milligrams per litre and often they are found only in few micrograms.
Standard Methods of Analysis
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. In many cases different methods
have been proposed for the same determination, and many of them were modified in some manner. As a result,
analytical data obtained by analysis were often in disagreement.
In an attempt to bring order out of chaos, the American Public Health Association appointed a committee to
study the various analytical methods available and published the recommendation of the committee as Standard
Methods of Water Analysis in 1905.
Standard Methods as published today is the product of the untiring effort of hundreds of individuals who
serve on committees, testing and improving analytical procedures for the purpose of selecting those best suited for
inclusion in Standard Methods, which is now available as Standard methods for the examination of water and
waste water.
1.0 EXPERIMENT ON DETERMINATION OF pH
PREAMBLE:
How to determine pH in Water and Wastewater.
Test procedure is in accordance to IS: 3025 (Part 11) - Reaffirmed 2002.
In addition to our Indian Standard, we also discuss in brief regarding the procedure
stated in
(1) APHA Standard Methods for the Examination of Water and Wastewater - 20
th
Edition. Method 4500-H
+
B.
(2) Methods for Chemical Analysis of Water and Wastes, EPA-600/4-79-020, USEPA,
Method 150.1.
1.1 AIM
To determine the PH of the given water sample with the stipulations as per IS: 3025
(Part 11) - Reaffirmed 2002
1.2 INTRODUCTION
The term pH refers to the measure of hydrogen ion concentration in a solution and
defined as the negative log of H
+
ions concentration in 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.
1.2.1 ENVIRONMENTAL SIGNIFICANCE
Determination of pH is one of the important objectives in biological treatment of the
wastewater. In anaerobic treatment, if the pH goes below 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 is
generally adjusted by addition of suitable acid or alkali to optimize the treatment of the
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 sludges, oxidation of cyanides and reduction of hexavalent chromium
into trivalent chromium also need a favorable pH range. It is used in the calculation of
carbonate, bicarbonate, CO
2
corrosion, stability index and acid base equilibrium.
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 are causing cancer in human beings.
1.3 PRINCIPLE
The pH electrode used in the pH measurement is a combined glass electrode. It
consists of sensing half cell and reference half cell, together form an electrode system.
The sensing half cell is a thin pH sensitive semi permeable membrane, separating two
solutions, viz., the outer solution, the sample to be analyzed and the internal solution,
enclosed inside the glass membrane and has a known pH value. An electrical
potential is developed inside and another electrical potential is developed outside, the
difference in the potential is measured and is given as the pH of the sample.
1.4 MATERIALS REQUIRED
1.4.1 APPARATUS REQUIRED
1. pH meter
2. Standard flasks
3. Magnetic Stirrer
4. Funnel
5. Beaker
6. Wash Bottle
7. Tissue Paper
8. Forceps
1.4.2 CHEMICALS REQUIRED
1. Buffers Solutions of pH 4.01, 7.0 and 9.2
2. Potassium Chloride
3. Distilled Water

1.5 SAMPLE HANDLING AND PRESERVATION
Preservation of sample is not practical. Because biological activity will continue after a
sample has been taken, changes may occur during handling and storage.

The characteristics of the water sample may change.

To reduce the change in samples taken for the determination of pH, keep samples at
4
0
C. Do not allow the samples to freeze.

Analysis should begin as soon as possible.

1.5.1 PRECAUTIONS
The following precautions should be observed while performing the experiment:
i. Temperature affects the measurement of pH at two points. The first is caused
by the change in electrode output at different temperatures. This interference
can be controlled by the instruments having temperature compensation or by
calibrating the electrode-instrument system at the temperature of the samples.
The second is the change of pH inherent in the sample at different
temperatures. This type of error is sample dependent and cannot be controlled;
hence both the pH and temperature at the time of analysis should be noted.
ii. In general, the glass electrode, is not subject to solution interferences like color,
high salinity, colloidal matter, oxidants, turbidity or reductants.
iii. Oil and grease, if present in the electrode layer, should be removed by gentle
wiping or detergent washing, followed by rinsing with distilled water, because it
could impair the electrode response.
iv. Before using, allow the electrode to stand in dilute hydrochloric acid solution
for at least 2 hours.
v. Electrodes used in the pH meter are highly fragile, hence handle it carefully.
1.6 PROCEDURE
Three major steps are involved in the experiment. They are
1. Preparation of Reagents
2. Calibrating the Instrument
3. Testing of Sample

1.6.1 PREPARATION OF REAGENTS
1. Buffer Solution of pH 4.0
Take 100 mL standard measuring flask and place a funnel over it.
Using the forceps carefully transfer one buffer tablet of pH 4.0 to the funnel.
Add little amount of distilled water, crush the tablet and dissolved it.
Make up the volume to 100 mL using distilled water.
Using the forceps carefully transfer one buffer tablet of pH 7.0 to the funnel.
Using the forceps carefully transfer one Buffer tablet of pH 9.2 to the funnel.
1.6.2 CALIBRATING THE INSTRUMENT
Using the buffer solutions calibrate the instrument.
Step 1
In a 100 mL beaker take pH 9.2 buffer solution and place it in a magnetic
stirrer, insert the teflon coated stirring bar and stir well.
Now place the electrode in the beaker containing the stirred buffer and check
for the reading in the pH meter.
If the instrument is not showing pH value of 9.2, using the calibration knob
adjust the reading to 9.2.
Take the electrode from the buffer, wash it with distilled water and then wipe
gently with soft tissue.

Step 2
1.6.3 TESTING OF SAMPLE
Step 3
Now the instrument is calibrated.
In a clean dry 100 mL beaker take the water sample and place it in a
magnetic stirrer, insert the teflon coated stirring bar and stir well.
Now place the electrode in the beaker containing the water sample and
check for the reading in the pH meter. Wait until you get a stable reading.
The pH of the given water sample is
Take the electrode from the water sample, wash it with distilled water and
then wipe gently with soft tissue.
1.7 CALCULATION
To determine the value of pH of the given water sample the readings obtained
are required to be tabulated
1.7.1 TABLE
Sample
No
Temperature of
Sample (C)
pH
1.
2.
3.
For sample 1 the temperature of the measurement is 27 C and as obtained the value
of the pH is
of the pH is
of the pH is
1.7.2 DATA SHEET
DETERMINATION OF pH
DATA SHEET
Date Tested :
Tested By :
Project Name :
Sample Number :
Sample Location BH1 :
Sample Description :
TABULATION
Sample
No
Temperature
of Sample
(C)
pH
1 27
2 27
3 27
Result:-
The pH of the given sample 1 =
1.8 INTERPRETATION OF RESULTS
The pH of the given water sample is
1.9 INFERENCE
pH is a measure of the hydrogen ion concentration in water. Values lower than 7
indicate acidity and values higher than 7 indicate alkalinity. Drinking water with a pH
between 6.5 and 8.5 is generally considered satisfactory. Acidic waters tend to be
corrosive to plumbing and faucets, particularly if the pH is below 6. Alkaline waters
are less corrosive. Waters with a pH above 8.5 may tend to have a bitter taste.
The pH of the water samples are well within the limit of the drinking water standards.
The pH of the ground water is slightly towards the alkaline side because of some soil
and rocks chemicals might have dissolved in it. In case of the pH of the fresh water,
aquatic plants uses up hydrogen molecules for photosynthesis, which causes the
concentration of hydrogen ions to decrease and therefore the pH is towards the
alkaline side. The sea water is mostly alkaline in nature because of the presence of
different type of salts.
1.10 EVALUATION
1. pH is defined as__________.
a) Logarithm of Hydrogen ions concentration
b) Negative logarithm of Hydrogen ions concentration
c) Hydrogen ion concentration
d) OH ion concentration
2. pH of neutral water is__________.
a) less than 7
b) more than 7
c) 7.0
d) 0.0
3. The acceptable value of pH of potable water is__________.
a) 7.0 to 8.5
b) 6.5 to 9.5
c) 6 to 8.5
d) 6.5 to 10
4. The inner solution present in the glass electrode of pH meter is__________.

a) HCl
b) KCl
c) NaCl
d) MgCL

5. The buffer solution can be stored for a minimum period at room temperature.
a) True
b) False

6. Possible reasons for a relatively low pH value in a river water sample is due to ___.

a) Organic material decomposition to form acidic substances
b) Running long distances
c) Presence of fishes
d) Presence of aquatic plants

7. Possible reasons for a relatively high pH value in a river water sample is due to ___.

a) Running over clay
b) Running long distances
c) Running of fishes
d) Presence of aquatic plants

8. A weak acid is one that ionize incompletely in aqueous solution.
a) True
b) False

9. A strong base is one that ionizes incompletely in aqueous solution.
a) True
b) False

10. The measurement of pH made by determining the e.m.f of the__________.
a) cell constant
b) solution
c) electrode cell
d) calomel electrode

KEY TO ITEMS:
1) b
2) c
3) a
4) b
5) False
6) a
7) d
8) True
9) False
10) c
2.0 EXPERIMENT ON DETERMINATION OF TURBIDITY
PREAMBLE:
How to determine turbidity in Water and Wastewater.
stated in
th
Edition. Method 2130 B.
Method 180.1.
2.1 AIM
To determine the turbidity of the given water sample with the stipulations as per IS:
3025 (Part 10) - Reaffirmed 2002.
2.2 INTRODUCTION
Turbidity is the technical term referring to the cloudiness of a solution and it is a
qualitative characteristic which is imparted by solid particles obstructing the
transmittance of light through a water sample. Turbidity often indicates the presence
of dispersed and suspended solids like clay, organic matter, silt, algae and other
microorganisms.
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 coloration is
apparent. The colloidal material which exerts turbidity provides adsorption sites for
chemicals and for biological organism that may not be harmful. They may be harmful
or cause undesirable tastes and odours. Disinfection of turbid water is difficult
because of 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 color 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. Knowledge of the turbidity
variation in raw water supplies is useful to determine whether a supply requires
special treatment by chemical coagulation and filtration before it may be used for a
public water supply. Turbidity measurements are used to determine the effectiveness
of treatment produced with different chemicals and the dosages needed. Turbidity
measurements help to gauge the amount of chemicals needed from day-to-day
operation of water treatment works.
Measurement of turbidity in settled water prior to filtration is useful in controlling
chemical dosages so as to prevent excessive loading of rapid sand filters. Turbidity
measurements of the filtered water are needed to check on faulty filter operation.
Turbidity measurements are useful to determine the optimum dosage of coagulants to
treat domestic and industrial wastewaters. Turbidity determination is used to evaluate
the performance of water treatment plants.
2.3 PRINCIPLE
Turbidity is based on the comparison of the intensity of light scattered by the sample
under defined conditions with the intensity of the light scattered by a standard
reference suspension under the same conditions. The turbidity of the sample is thus
measured from the amount of light scattered by the sample taking a reference with
standard turbidity suspension. The higher the intensity of scattered light the higher is
the turbidity. Formazin polymer is used as the primary standard reference
suspension.
1. Turbidity Meter
2. Sample Cells
3. Standard flasks
4. Funnel
5. Wash Bottle
6. Tissue Papers

1. Hexamethylenetetramine
2. Hydrazine sulphate
3. Distilled water

Water samples should be collected in plastic cans or glass bottles. All bottles must be
cleaned thoroughly and should be rinsed with turbidity free water.
Volume collected should be sufficient to insure a representative sample, allow for
replicate analysis (if required), and minimize waste disposal.
No chemical preservation is required. Keep the samples at 4C. Do not allow
samples to freeze.
Analysis should begin as soon as possible after the collection. If storage is required,
samples maintained at 4C may be held for up to 48 hours.
2.5.1 PRECAUTIONS
The presence of coloured solutes causes measured turbidity values to be low.
Precipitation of dissolved constituents (for example, Fe) causes measured
turbidity values to be high.
Light absorbing materials such as activated carbon in significant
concentrations can cause low readings.
The presence of floating debris and coarse sediments which settle out rapidly
will give low readings. Finely divided air bubbles can cause high readings.
2.6 PROCEDURE
For testing the given water sample first the reagents are to be prepared. Then the
turbidity meter is required to be calibrated.
1. Hydrazine Sulphate
Weigh accurately 1 g of hydrazine sulphate and dissolve it in turbidity free
distilled water.
Transfer it to a 100 mL standard flask and make up to 100 ml using turbidity
free distilled water.
2. Hexamethylene Tetramine
Weigh accurately 10 g of Hexamethylene tetramine and dissolve it in turbidity
Transfer it to a 100 mL standard flask and make up to 100 ml using turbidity
3. Standard 4000 NTU Solution
Mix 5 mL of hydrazine sulphate solution and 5 mL of Hexamethylenetetramine
solution in a 100 mL standard measuring flask.
Allow the mixture to stand for 24 hours.
After 24 hours, make up the volume to 100 mL using turbidity free distilled
water.
The standard 4000 NTU solution is ready.
2.6.2 CALIBRATION OF TURBIDITY METER
Using the standard solution calibrate the instrument.
The instrument is having four knobs, out of which the two knobs in the bottom is the
set zero knob, this is for setting the instrument to zero.
The one which is there in the top left hand side is the calibration knob, used for the
calibration.
The other one in the top is the knob for setting the detection range. It is adjusted to
1000 NTU range.
Step 1
To the sample cells, add turbidity free distilled water up to the horizontal mark, wipe
gently with soft tissue. Place it in the turbidity meter such that the vertical mark in the
sample cell should coincide with the mark in the turbidity meter and cover the sample
cell. Now using the set zero knob, adjust the reading to zero.
Step 2
According to our need, prepare a standard solution. In this case, a 200 NTU solution
is prepared by diluting the standard 4000 NTU solution and added to the sample cells,
up to the horizontal mark, wipe gently with soft tissue. Place it in the turbidity meter
such that the vertical mark in the sample cell should coincide with the mark in the
turbidity meter and cover the sample cell.
If the instrument is not showing 200 NTU, using the calibration knob adjust the
reading to 200 NTU.
Repeat the procedure for two / three times.
Now the instrument is calibrated.

2.6.3 TESTING OF WATER SAMPLE
To the sample cells, add sample water up to the horizontal mark, wipe gently
with soft tissue and place it in the turbidity meter such that the vertical mark in
the sample cell should coincide with the mark in the turbidity meter and cover
the sample cell.
Check for the reading in the turbidity meter. Wait until you get a stable reading.
The turbidity of the given water sample is 8.4 NTU.
2.7 CALCULATION
For determining the Turbidity of the given water sample the readings are required to
be tabulated.
2.7.1 TABLE
Sample
No.
Temperature of
Sample (C)
Turbidity
(NTU)
1.
2.
3.
For sample 1 the temperature of the sample is 27C and turbidity value NTU
For sample 2 the temperature of the sample is 27C and the turbidity value NTU
For sample 3 the temperature of the sample is 27C and obtained turbidity value is
NTU
2.7.2 DATA SHEET
DETERMINATION OF TURBIDITY
DATA SHEET
Date Tested :
Tested By :
Project Name :
Sample Number :
TABULATION
Sample
No
Temperature
of Sample
(C)
Turbidity
(NTU)
1 27
2 27
3 27
Result:-
The turbidity of the given sample 1 =
The turbidity of the given water sample is 8 NTU.
[
2.9 INFERENCE

Turbidity is a measure of light transmission and indicates the presence of suspended
material such as clay, silt, finely divided organic material, plankton and other
inorganic material. If turbidity is high, be aware of possible bacterial contamination.
Normally the ground water is clear in nature and it will satisfy the codes need. The
ground water may get contaminated by intrusion of domestic or industrial wastewater
causing turbidity of the sample. Turbidity in excess of 5 NTU is usually objectionable
for aesthetic reasons. In case of freshwater lakes and ponds, due to contamination
and algal growth the turbidity of these water increases to very high levels. The clarity
of sea water is very low because of huge amount of suspended particles, thereby
increasing the turbidity.

2.10 EVALUATION
1. Turbidity is caused by Clay, Silt, Organic matter and Microbes.
a) True
b) False

2. The turbidity is measured based on the

a) Light absorbing properties
b) Light Scattering properties
c) Particle Size
d) Particle mass

3. The colour of the water sample affects the turbidity.
a) True
b) False

4. In a nephelo turbidity meter the light detectors are at
a) 180
b) 360
c) 90
d) 270

5. What is the unit of turbidity.

a) TU
b) MTU
c) NTU
d) IU

6. What is the light source for the nephelo turbidity meter?
a) Tungsten filament lamp
b) Deuterium lamp
c) Hallow Cathode lamp
d) Sodium vapour lamp

7. The turbidity affects the aquatic life in the water.
a) True
b) False

8. The standard unit of turbidity is considered as that produced by
a) 2ppm of silica in distilled water
b) 1ppm of silica in distilled water
c) 4ppm of silica in distilled water
d) 9ppm of silica in distilled water

9. The material used in the standard solution for nephelometer is ___.

a) Silica
b) Clay
c) Formazin
d) Barium Chloride

10. Mixture of hydrazine sulphate and hexamethylenetetramine solution is allowed to
stand for _____.

a) 24 hours
b) 12 hours
c) Minimum 6 hours
d) No specific time

KEY TO ITEMS:

1) True
2) b
3) True
4) c
5) c
6) a
7) True
8) b
9) c
10) a

3.0 EXPERIMENT ON DETERMINATION OF CONDUCTIVITY
PREAMBLE:
How to determine conducti vity in Water and Wastewater.
In addition to our Indian Standard, we also discuss in brief regarding the
procedure stated in
th
Edition. Method 2510.
(2) Methods for Chemical Analysis of Water and Wastes, EPA-600/4-79-020,
USEPA, Method 120.1.
3.1 AIM
To determine the conductivity of given water sample with the stipulations as per
IS: 3025 (Part 14) - Reaffirmed 2002.
3.2 INTRODUCTION
Conductivity of a substance is defined as 'the ability or power to conduct or
transmit heat, electricity or sound'. When an electrical potential difference is
placed across a conductor, its movable charges flow, giving rise to an electric
current. This property is called conductivity. Since the charge on ions in solution
facilitates the conductance of electrical current, the conductivity of a solution is
proportional to its ion concentration.
The electrical conductivity can be expressed as mhos (Reciprocal of ohms) or as
siemens. The conductivity of water is a measure of the ability of water to carry an
electric current. In most water, the conductivity is very low, so millisiemens or
microsiemens are used as units for water conductivity. The conductivity of water
is directly linked to the concentration of the ions and their mobility. The ions in
water acts as electrolytes and conducts the electricity.
The conductivity depends on the value of the pH, on the temperature of
measurement and on the amount of CO
2
which has been dissolved in the water
to form ions. The conductivity is also affected by the concentration of ions
already present in the water such as chloride, sodium and ammonium. Chemical
composition of water determines its conductivity. Hence this becomes the most
widely used measure of the purity of water.
.3.2.1 ENVIRONMENTAL SIGNIFICANCE
Electrical conductivity measurements are often employed to monitor
desalination plants.
It is useful to assess the source of pollution.

In coastal regions, conductivity data can be used to decide the extent of
intrusion of sea water into ground water.
Conductivity data is useful in determining the suitability of water and
wastewater for disposal on land. Irrigation waters up to 2 millisiemens / cm
conductance have been found to be suitable for irrigation depending on
soils and climatic characteristics.

It is also used indirectly to fine out inorganic dissolved solids.
3.3 PRINCIPLE
Conductivity is measured with a probe and a meter. A voltage is applied between
the two electrodes in the probe immersed in the sample water. The drop in
voltage caused by the resistance of the water is used to calculate the
conductivity per centimeter.

Conductivity (G), the inverse of resistivity (R) is determined from the voltage and
current values according to Ohms law. i.e. R=V/I then, G=1/R=I/V.

The meter converts the probe measurement to micro mhos per centimeter and
displays the result for the user.

1. Conductivity Meter with Electrode /ATC probe
2. Magnetic Stirrer with stirring bead
3. Standard flask
4. Measuring jar
5. Beaker 250 mL
6. Funnel
7. Tissue Paper

1. Potassium Chloride
2. Distilled Water

Water samples should be collected in plastic cans or glass bottles. All bottles
must be cleaned thoroughly in phosphate-free detergent and rinsed thoroughly
with both tap and distilled water.

Volume collected should be sufficient to insure a representative sample, allow for
replicate analysis (if required), and minimize waste disposal.
Analysis should begin as soon as possible after the collection. If the analysis is
not completed within 12 hours of sample collection, sample should be filtered
through a 0.45 filter paper and stored at 4C. High quality distilled water must
be used for washing the filter and apparatus and needs to be rinsed with sample
before use.

No chemical preservation is required. Keep the samples at 4C. DO NOT
ALLOW SAMPLES TO FREEZE.

3.5.1 PRECAUTIONS
1. Switch on the conductivity meter for atleast 30 minutes before starting the
experiment so that the instrument gets stabilizes.
2. As it involves instruments for analyzing do not forget to calibrate the
instrument.
3. Always prepare the calibration solution freshly before the start of the
experiment.
4. As conductance is influenced by temperature, always use a conductivity
meter with temperature control.
5. Always dip the electrode in distilled water and do not expose it to air.
3.6 PROCEDURE
For testing the given water sample first the reagents are to be prepared. Then
the Conductivity Meter is required to be calibrated.
Potassium Chloride Solution (0.1N):
Switch on the Electronic balance, keep the weighing pan, set the reading
to zero.

Measure 50 mL of distilled water and transfer it to the beaker.
Weigh 0.7456g of Potassium chloride.
Transfer the 0.7456g of potassium chloride to the beaker contains distilled
water and mix it by the glass rod until it dissolves thoroughly.
Transfer the contents to the 100 mL standard flask.
Make up the volume to 100 mL, by adding distilled water and shake the
contents well. This solution is used to calibrate the conductivity meter.
1. Conductivity Meter
An overview on conductivi ty meter:
An electrical conductivity meter is used to measure the conductivity in a solution.
Basically the conductivity unit consists of
2. Electrode / ATC probe
3. Magnetic stirrer with bead
Before starting the experiment, switch on the instrument for atleast 30 minutes,
so that the instrument stabilizes. Using the same electrode the Conductivity
meter can measure three parameters
1. Conductivity
2. Selenity
3. Total Dissolved Solids (TDS)
The Room temperature can also be displayed using Automatic temperature
control (ATC) probe. The Mode button is pressed to select the parameter to be
measured. The Display can show conductivity reading in microsiemens or in
millisiemens. The Conductivity of a solution is highly influenced by temperature
therefore it is necessary to calibrate the instrument at the same temperature as
the solution is being measured.
Take 0.1N Potassium Chloride in a beaker. Switch on the magnetic stirrer and
place the beaker on the stirrer. Insert the magnetic bead in the beaker. Place the
electrode inside the solution. Select the calibration button and using up and
down key adjust the conductivity of the 0.1N potassium chloride solution to 14.12
millisiemens / cm at 30
o
c. Now the meter is ready for the measurement of
samples.
Calibration of Conductivity Meter
Rinse the electrode thoroughly with deionised water and carefully wipe
with a tissue paper.
Measure 200 mL of water sample and transfer it to a beaker and place it
on the magnetic stirrer.
Dip the electrode into the sample solution taken in a beaker and wait for a
steady reading. Make sure that the instrument is giving stable reading.
Note down the reading in the display directly, which is expressed in
millisiemens.
The reading is millisiemens.
3.7 DATA SHEET
DETERMINATION OF CONDUCTIVITY
DATA SHEET
Date Tested :
Tested By :
Project Name :
Sample Number :
TABULATION
Sample
No
Temperature
of Sample (C)
Conductivity
(mho)
1. 27
2. 27
3. 27
Result:-
The conductivity of the given sample 1 =
The conductivity of the given water sample is millisiemens.
3.9 INFERENCE
The conductivity value gives us a rapid and inexpensive way of determining the
ionic strength of a solution. This is an easy measurement to make and relates
closely to the total dissolved solids content of water. The total dissolved solids
are about seventy percent of the conductivity. In the ground water, the ionisable
salts are lesser and thereby the conductivity is also lesser in nature. Water
having more number of ionisable salts for example sea water, is having high
conductivity. The fresh water bodies only have a minimum amount of salts and
have moderate conductivity.
Solution S/cm
Totally pure water 0.055
Typical DI water 0.1
Distilled water 0.5
RO water 50-100
Domestic "tap" water 500-800
Potable water (max) 1055
Sea water 56,000
Brackish water 100,000
3.10 EVALUATION

1. Conductivity is the measure of the ability of water to carry the ions.
a) True
b) False

2. The unit of conductance is _____ .

a) mho
b) ohm
c) ampere
d) watts

3. The conductivity of standard 0.01M KCl solution is

a) 1412 mhos/ cm
b) 1412 mmhos/ cm
2

c) 1412 mhos/ mm
d) 1412 mmhos/ mm
2

4. The conductivity of a sample depends on Temperature.
a) True
b) False

5. Using a conductivity meter we can measure _______ of the solution.
a) specific conductance
b) equivalent conductance
c) Specific resistance
d) concentration

6. The Conductivity is the maximum for _______ water.

a) Distilled
b) Deionized
c) Ground
d) Sea

7. The conductivity of potable water varies from
a) 2501 to 5000 mhos /cm
b) 50 to 1500 mhos /cm
c) 1501 to 2000 mhos /cm
d) 2001 to 2500 mhos /cm

8. The measurement of conductivity may lead to the estimation of ___.

a) Total solids
b) Total dissolved solids
c) Colloidal solids
d) Suspended solids

9. Freshly made distilled water has a conductivity of
a) 2.0 to 3.0 mhos /cm
b) 0.5 to 2.0 mhos /cm
c) 2.5 to 4.5 mhos /cm
d) 4.5 to 5.0 mhos /cm

10. The conductance of a solution placed between two electrodes of 1 cm2 area
& kept 1 cm apart is Molar conductance.

a) True
b) False

KEY TO ITEMS:

1) False
2) a
3) a
4) True
5) a
6) d
7) b
8) b
9) b
10) False
4.0 EXPERIMENT ON DETERMINATION OF TOTAL DISSOLVED
AND SUSPENDED SOLIDS IN WATER
PREAMBLE:
How to determine total dissolved and suspended solids in Water and
Wastewater.
Test procedure is in accordance to IS: 3025 (Part 16 & Part 17).
procedure stated in
th
Edition. Method 2540 C and 2540 D.
Method 160.1.
4.1 AIM
To determine total dissolved and suspended solids in the given water sample
with the stipulations as per IS: 3025 (Part 16 & Part 17).
4.2 INTRODUCTION
The term total dissolved solids refer to materials that are completely dissolved in
water. These solids are filterable in nature. It is defined as residue upon
evaporation of filterable sample. The term total suspended solids can be referred
to materials which are not dissolved in water and are non filterable in nature. It is
defined as residue upon evaporation of non filterable sample on a filter paper.
Dissolved minerals, gases and organic constituents may produce
aesthetically displeasing colour, taste and odour.
Some dissolved organic chemicals may deplete the dissolved oxygen in
the receiving waters and some may be inert to biological oxidation, yet
others have been identified as carcinogens.
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. In industries, the use of water with high amount of
dissolved solids may lead to scaling in boilers, corrosion and degraded
quality of the product.
Estimation of total dissolved solids is useful to determine whether the
water is suitable for drinking purpose, agriculture and industrial purpose.
Suspended material is aesthetically displeasing and provides adsorption
sites for chemical and biological agents.
Suspended organic solids which are degraded anaerobically may release
obnoxious odours.
Biologically active suspended solids may include disease causing
organisms as well as organisms such as toxic producing strains of algae.
The suspended solids parameter is used to measure the quality of
wastewater influent and effluent.
Suspended solids determination is extremely valuable in the analysis of
polluted waters.
Suspended solids exclude light, thus reducing the growth of oxygen
producing plants.
4.3 PRINCIPLE
A well mixed sample is filtered through a standard glass fiber filter, and the filtrate
is evaporated to dryness in a weighed dish and dried to constant weight at 179-
181C. The increase in dish weight represents the total dissolved solids.
A well mixed sample is filtered through a weighed standard glass fiber filter and
the residue retained on the filter is dried to a constant weight at 103-105C. The
increase n weight of the filter represents the total suspended solids. If the
suspended material clogs the filter and prolongs filtration, the difference between
the total solids and total dissolved solids may provide an estimate of the total
suspended solids.
1. Evaporating Dish
2. Water Bath
3. Oven
4. Desiccators
5. Analytical Balance
6. Graduated Cylinders
7. Dish Tongs
8. Gooch Crucibles
9. Filter
10. Vacuum Pumps
11. Crucible tongs
12. Forceps, Smooth -tipped

Preservation of sample is not practical. Because biological activity will continue
after a sample has been taken, changes may occur during handling and storage.
Both the characteristics and the amount of solids may change.
To reduce this change in samples taken for solids determinations, keep all
samples at 4
0
C.
Do not allow samples to freeze.
4.5.1 PRECAUTIONS
Water or Wastewater samples which contain high concentrations of
calcium, chloride, magnesium or sulfate can rapidly absorb moisture from
the air.
Such samples may need to be dried for a longer period of time, cooled
under proper desiccation and weighed rapidly in order to achieve a
reasonable constant weight.
We should be aware prolonged drying may result in loss of constituents,
particularly nitrates and chlorides.
Volume of sample should be adjusted to have residue left after drying as
100 to 200mg. It is mainly to prevent large amount of residue in entrapping
water during evaporation.
Samples with high concentrations or bicarbonate require additional drying
at 180C to ensure that all of the bicarbonate is converted to carbonate.
4.6 PROCEDURE
4.6.1. TESTING OF SAMPLE FOR TOTAL DISSOLVED SOLIDS
To measure total dissolved solids, take a clean porcelain dish which has been
washed and dried in a hot air oven at 180(C for one hour.
Now weigh the empty evaporating dish in analytical balance. Lets denote
the weight measured as W1 = g.
Mix sample well and pour into a funnel with filter paper. Filter
approximately 80 -100 mL of sample.
Using pipette transfer 75mL of unfiltered sample in the porcelain dish.
Switch on the oven and allowed to reach 105C. Check and regulate oven
and furnace temperatures frequently to maintain the desired temperature
range.
Place it in the hot air oven and care should be taken to prevent splattering
of sample during evaporation or boiling.
Dry the sample to get constant mass. Drying for long duration usually 1 to
2 hours is done to eliminate necessity of checking for constant mass.
Cool the container in a desiccator. Desiccators are designed to provide
an environment of standard dryness. This is maintained by the desiccant
found inside. Don't leave the lid off for prolonged periods or the desiccant
will soon be exhausted. Keep desiccator cover greased with the
appropriate type of lubricant in order to seal the desiccator and prevent
moisture from entering the desiccator as the test glassware cools.
We should weigh the dish as soon as it has cooled to avoid absorption of
moisture due to its hygroscopic nature. Samples need to be measured
accurately, weighed carefully, and dried and cooled completely.
Note the weight with residue as W
2
= g.
4.6.2 TESTING OF SAMPLE FOR TOTAL SUSPENDED SOLIDS
Place filtration apparatus with weighed filter in filter flask.
Mix sample well and pour into a graduated cylinder to the selected
volume.
Apply suction to filter flask and seat filter with a small amount of distilled
water.
Pour selected volume into filtration apparatus.
Draw sample through filter into filter flask.
Rinse graduated cylinder into filtration apparatus with three successive 10
mL portions of distilled water, allowing complete drainage between each
rinsing.
Continue suction for three minutes after filtration of final rinse is
completed.
Dry filter in an oven at 103-105C for at least 1 hour.
Cool filter in desiccator to room temperature.
When cool, weigh the filter and support.
4.7 CALCULATION
4.7.1 TABLE
Total Dissolved Solids
Tabulation for Total Dissolved Solids (TDS):
Weight of the clean porcelain evaporating dish (g) W
1
=
Weight of the dish and the residue (g) W
2
=
Weight of residue (g) W =
The volume of the sample (mL) V =
Description Weight (g)
Weight of the clean porcelain evaporating dish (g) W1
Weight of the dish and the residue (g) W2
Weight of residue(g) W
Volume of the Sample (mL) V
Total Dissolved Solids (mg/L) TDS
Total Suspended Solids
Tabulation for Total Suspended Solids (TSS)
Weight of the clean filter paper (g) W
1
=
Weight of the clean filter paper and the residue (g) W
2
=
Weight of residue (g) W =
Volume of the sample (mL) V =
1

Weight of the filter paper and the residue (g) W
2

Total Suspended Solids (mg/L) TSS
4.7.2 DATA SHEET
DETERMINATION OF TOTAL DISSOLVED SOLIDS
DATA SHEET
Date Tested :
Tested By :
Project Name :
Sample Number :
Sample Location :
Model Calculation:
W1 =
W2 =
V =
Weight of residue (g) W = W2 -W1
=
=
Weight of residue in mg (To convert W (g) to W (mg), multiply W (g) with
1000)
W (mg) =
Multiply the weight of the dry solids (in mg) by 1,000 mL/L to convert the sample size from mL to L.
Description
Total Dissolved Solids (mg/L)
V = Volume of the sample (mL) (To convert mL to L, multiply by
1000)
Weight
(g)
Weight of the clean porcelain evaporating dish (g) W
1
Weight of the dish and the residue (g) W
2
Total Dissolved Solids (mg/L) TDS
DETERMINATION OF TOTAL SUSPENDED SOLIDS
DATA SHEET
Date Tested :
Tested By :
Project Name :
Sample Number :
Sample Location :
Model Calculation:
W1 =
W2 =
V =
=

1000)
W (mg) =
= 20.2 mg
Description
Total Suspended Solids (mg/L)
1000)

Weight
(g)
1
Weight of the filter paper and the residue (g) W
2
Total Suspended Solids (mg/L) TDS
In the given sample, total dissolved solid is to mg/L and total suspended
solid is to mg/L.
4.9 INFERENCE
Water can be classified by the amount of TDS per litre:
fresh water

<1500 mg/L TDS
brackish water

1500 to 5000 mg/L TDS
saline water
The following charts give some common ranges for TSS results and possible
removal efficiencies for various types of treatment.
>5000 mg/L TDS
Sample Common Ranges, mg/L
Influent Weak <150 400+Strong
Primary Effluent Weak <60 150+Strong
Secondary Effluent Good 10 - 60+Bad
Tertiary Effluent Less than 3
Activated Sludge
Mixed Liquor (MLSS) 1,000 - 5,000
Return or waste sludge 2,000 - 12,000
Digester Supernatent 3,000 - 10,000
Sludge 20,000 - 60,000
4.10 EVALUATION
1. The pore size of the filter paper used for filtration is
a) 2.0m or smaller
b) 2.0m or bigger
c) 2.0m
d) 20.0m
2. The type of crucible used for the experiment is made of _______.

a) Porcelain
b) Clay
c) Silver
d) Iron

3. Total Suspended Solids are mostly responsible for

a) Turbidity.
b) colour
c) Odour
d) Taste

4. The chemical substance used in the desiccators is _____.

a) Calcium Chloride
b) Calcium Carbonate
c) Sodium Chloride
d) Sodium Hydroxide

5. Always the Total Suspended Solids value will be
a) Less than Total Dissolved Solids
b) Greater than Total Dissolved Solids
c) Less than Total Solids
d) Greater than Total Solids

6. High total dissolved solids indicates lower level of hardness.
a) True
b) False

7. The concentration of dissolved solids in water can be determined by specific
conductance.

a) True
b) False

8. The settleable suspended solids with diameter 0.15 to 0.2mm are generally

a) inorganic
b) Organic
c) algae
d) fungi

9. The dissolved solids that impose BOD are ______.

a) volatile solids
b) non volatile solids
c) inorganic solids
d) total solids

10. As per IS Code the acceptable TDS value is

a) 250 ppm
b) 500 ppm
c) 750 ppm
d) 900 ppm

KEY TO ITEMS:

1) a
2) a
3) a
4) a
5) c
6) False
7) True
8) a
9) a
10) b

5.0 EXPERIMENT ON DETERMINATION OF ALKALINITY OF
WATER
PREAMBLE:
How to determine alkalinity in Water and Wastewater.
procedure stated in
th
5.1 AIM
To determine the alkalinity of given water sample with the stipulations as per
5.2 INTRODUCTION
Alkalinity is primarily a way of measuring the acid neutralizing capacity of water.
In other words, its ability to maintain a relatively constant pH.
The possibility to maintain constant pH is due to the hydroxyl, carbonate and
bicarbonate ions present in water.
The ability of natural water to act as a buffer is controlled in part by the amount of
calcium and carbonate ions in solution.
Carbonate ion and calcium ion both come from calcium carbonate or limestone.
So water that comes in contact with limestone will contain high levels of both
Ca
++
and CO
3
2-
ions and have elevated hardness and alkalinity.
Alkalinity is important for fish and aquatic life because it protects or buffers
against rapid pH changes. Higher alkalinity levels in surface waters will buffer
acid rain and other acid wastes and prevent pH changes that are harmful to
aquatic life.
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 waters. The resultant precipitate can corrode
pipes and other accessories of water distribution systems.
Wastewaters containing excess caustic (hydroxide) alkalinity are not to be
discharged into natural water bodies or sewers.
Alkalinity as carbonate and bicarbonate of saline water is very important in
tertiary recovery processes for recovering petroleum. Alkaline water offers better
wetting to the formation rock and improve oil release. As an additional benefit,
ions that provide alkalinity absorb on rock surfaces occupying adsorption sites
and decrease the loss of recovery chemical by adsorption.
The alkalinity value is necessary in the calculation of carbonate scaling
tendencies of saline waters.
The alkalinity acts as a pH buffer in coagulation and lime-soda softening of water.
In wastewater treatment, alkalinity is an important parameter in 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 become considerations.
5.3 PRINCIPLE
The alkalinity of water can be determined by titrating the water sample with
Sulphuric acid of known values of pH, volume and concentrations. Based on
stoichiometry of the reaction and number of moles of Sulphuric acid needed to
reach the end point, the concentration of alkalinity in water is calculated.
When a water sample that has a pH of greater than 4.5 is titrated with acid to a
pH 4.5 end point, all OH
-
, CO
3
2-
, and HCO
3
-
will be neutralized.
For the pH more than 8.3, add phenolphthalein indicator, the colour changes to
pink colour. This pink colour is due to presence of hydroxyl ions.
If sulphuric acid is added to it, the pink colour disappears i.e. OH
-
ions are
neutralized.
Then add mixed indicator, the presence of CO
3
2-
and HCO
3
-
ions in the solution
changes the colour to blue. While adding sulphuric acid, the color changes to red,
this color change indicates that all the CO
3
2-
and HCO
3
-
ions has been
neutralized. This is the end point.
1. Burette with Burette stand and porcelain title
2. Pipettes with elongated tips
3. Pipette bulb
4. Conical flask (Erlenmeyer Flask)
5. 250 mL Measuring cylinders
6. Standard flask
7. Wash Bottle
8. Beakers
1. Standard sulphuric acid
2. Phenolphthalein
3. Mixed Indicator
4. Bromocresol Green
5. Methyl Red
6. Ethyl alcohol
7. Distilled Water

To reduce the change in samples, keep all samples at 4C. Do not allow samples
to freeze.
Do not open sample bottle before analysis.
5.5.1 PRECAUTIONS
The following precautions should be observed while performing the
experiment:
1. Do not keep the indicator solution open since it contains the alcohol which
tends to evaporate.
2. The mixed indicator solution is containing dye in it; care should be taken
so that it is not spilled to your skin.
3. If it spills on your skin, the scar will remain at least for two to three days.
5.6 PROCEDURE
For testing the given sample, first the reagents are required to be prepared.
Take approximately 500 mL of distilled water in a 1000 mL standard flask.
Sulphuric Acid Solution (0.02N):
Pipette 20 mL of concentrated 0.1 Normality Sulphuric acid and add slowly
along the sides of the standard flask.
Then make up the volume up to 1000 mL mark. Now the strength of this
solution is 0.02 N.
Weigh 1g of phenolphthalein and add to 100 mL of 95% ethyl alcohol or to
100 mL of distilled water. Use the readymade Phenolphthalein indicator
available in the market.
Phenolphthalein Indicator Preparation:
Dissolve 100 mg Bromocresol green and 20 mg of methyl red in 100 mL of
95% ethyl alcohol or use 100 mL of distilled water. Mixed indicator also
readily available in the market. So it can be used as indicator in this
experiment.
Mixed Indicator Preparation:
Rinse the burette with 0.02N Sulphuric acid and discard the solution.
Fill the burette with 0.02N sulphuric acid and adjust it to zero.
Fix the burette in the stand.
Using a measuring cylinder exactly measure 100 mL of sample and pour it
into a 250 mL of conical flask.
Add few drops of phenolphthalein indicator to the contents of conical flask.
The colour of the solution will turn to pink. This colour change is due to
alkalinity of hydroxyl ions in the water sample.
Titrate it against 0.02N sulphuric acid till the pink color disappears. This
indicates that all the hydroxyl ions are removed from the water sample.
Note down the titter value (V1). The value of titration is 0.5mL .This value
is used in calculating the phenolphthalein alkalinity.
To the same solution in the conical flask add few drops of mixed indicator.
The colour of the solution turns to blue. This colour change is due to CO
3
2-

& HCO
3
-
ions in water sample.
Continue the titration from the point where stopped for the phenolphthalein
alkalinity. Titrate till the solution becomes red. The entire volume (V2) of
sulphuric acid is noted down and it is accountable in calculating the total
alkalinity.
The value of titration is 8.3mL.
Repeat the titration for concordant values.
5.7 CALCULATION
5.7.1 TABLE
Table -1 Phenolphthalein Alkalinity:
Sl.No.
Volume of
Sample (mL)
Burette Reading (mL)
Volume of
Sulphuric acid
(mL)
Initial Final
1.
2.
3.
Burette solution: Sulphuric Acid Solution
Pipette solution: Sample.
Indicator: Phenolphthalein Indicator.
End point: Disappearance of pink color.
For the calculation of Phenolphthalein Alkalinity
The Sulphuric acid is taken in the Burette.
For the first titration, the volume of water sample taken is 100 mL. The
initial reading is . The final reading is .
The volume of sulphuric acid consumed to get the end point is mL.
For the second titration, the volume of water sample taken is 100 mL. The
initial reading is _____ The final reading is _____.
The volume of sulphuric acid consumed to get the end point is ____ mL.
For the third titration, the volume of water sample taken is 100 mL. The
initial reading is ____. The final reading is _____.
The volume of Sulphuric Acid consumed to get the end point is _____ mL.
For the second and third titration, the burette reading is same so we have
achieved concordant value. We can go for the calculations
Phenolphthalein Alkalinity of the given water sample is equal to volume of
H
2
SO
4
(V
1
) * normality * 50 *1000 divided by volume of sample taken
Here the volume of H
2
SO
4
(V
1
) is _____mL
Normality is _______Molar
And volume of sample taken is 100mL
Substituting the values in the formula and calculating we get the value
______ mg/L
So the Phenolphthalein Alkalinity mg/L is _______ mg/L
Table - 2 Total Alkalinity:
Burette solution: Sulphuric Acid Solution
Pipette solution: Sample.
Indicator: Mixed Indicator.
End point: Appearance of Red color.
For Calculation of Total Alkalinity the volume of water sample taken is 100
mL.
The Sulphuric acid is taken in the Burette.
For the first titration, the volume of water sample taken is 100 mL. The
initial reading is _______.The final reading is _____.
The volume of sulphuric acid consumed to get the end point is _____ mL.
For the second titration, the volume of water sample taken is 100 mL.
The initial reading is _______.The final reading is ______.
The volume of sulphuric acid consumed to get the end point is _____mL.
For the third titration, the volume of water sample taken is 100 mL. The
initial reading is ______. The final reading is _______
The volume of H
2
SO
4
(V
2
) consumed to get the end point is ______ mL.
For the second and third titration, the burette reading is same so we have
achieved concordant value. We can go for the calculations
Total Alkalinity of the given water sample is equal to volume of H
2
SO
4
(V
2
)
* normality * 50 * 1000 divided by volume of sample taken
Here the volume of H
2
SO
4
(V
2
) is _______ mL.
Normality is _________Molar and volume of sample taken is 100mL.
Substituting the values in the formula and calculating we get the value
____mg/L. So the Total Alkalinity is _______ mg/L.
Sl.No.
Volume of
Sample (mL)
Volume of
Sulphuric acid
(mL)
Initial Final
1.
2.
3.
7.7.2 DATA SHEET
DETERMINATION OF ALKALINITY
DATA SHEET
Date Tested :
Tested By :
Project Name :
Sample Number :
Sample Location :
Phenolphthalein Alkalinity =
Specimen Calculation:
Volume of Sulphuric Acid =
Normality of Sulphuric Acid =
Volume of Sample =
Equivalent weight of CaCO3 =
(volume of H
2 2SO4(v1)* Normality * 50 * 1000)
Volume of sample taken
To convert the sample size from mL to L
Sl.No.
, multiply the result by 1,000 mL/L to convert the sample size
from mL to L
Alkalinity as CaCO3 equivalent (mg/L) =
= mg/L as CaCO
3
equivalent
Volume of
Sample (mL)
Burette Reading (mL) Volume of
Sulphuric acid (mL)
Initial Final
4. 100
5. 100
6. 100
Total Alkalinity:
Total Alkalinity =
Volume of Sulphuric Acid =
Volume of Sample =
(volume of H
2
2SO4(v1)* Normality * 50 * 1000)
To convert the sample size from mL to L, multiply the result by 1,000 mL/L to convert the sample size
from mL to L
Alkalinity as CaCO3 equivalent (mg/L) =
=
Sl.No.
Volume of
Sample (mL)
Sulphuric acid (mL)
Initial Final
1 100
2 100
3 100
Value of P and T
Al kalinity due to
OH
-
CO
3
2-
HCO
3
-

P=0 0 0 T =83
P< T 0 2P =10 T-2P =73
P= T 0 2P =10 0
P> T 2P-T =- 73 2P-T =- 73 0
P=T T =83 0 0
To find the different values of Alkalinity due to Hydroxyl, Carbonate and
Bicarbonate ions take P as Phenolphthalein Alkalinity and T as Total
Alkalinity.
If P=0, The Alkalinity due to Hydroxyl and carbonate ions is 0. Alkalinity
due to Bicarbonate ion is equal to the Total Alkalinity i.e. T =______ mg/L.
If P < ,T then the Alkalinity due to Hydroxyl ion is 0. The Alkalinity due to
carbonate ion is 2P. i.e. 2P =______ mg/L. Alkalinity due to Bicarbonate
ion is equal to the Total Alkalinity minus 2 times Phenolphthalein
Alkalinity
i.e. T-2P =______ mg/L.
If P = ,T then the Alkalinity due to Hydroxyl ion is _____. The Alkalinity
due to carbonate ion is 2P. i.e. 2P =______ mg/L. Alkalinity due to Bicar-
bonate ion is equal to ______
If P > ,T then the Alkalinity due to Hydroxyl and carbonate ions is 2P
T. i.e. 2P-T =______ mg/L. Alkalinity due to Bicarbonate ion is ______
If P=T, The Alkalinity due to Hydroxyl is equal to the Total Alkalinity i.e. T
=______ mg/L. Alkalinity due to carbonate and Bicarbonate ions is ____.
If P >, T then the Alkalinity due to Hydroxyl and carbonate ions is 2PT.
i.e. 2P-T =____ mg/L. Alkalinity due to Bicarbonate ion is 0. If P =T, The
Alkalinity due to Hydroxyl is equal to the Total Alkalinity. i.e. T =___ mg/L.
Alkalinity due to carbonate and Bicarbonate ions is 0.
5.9 INFERENCE
Alkalinity is a measure of the capacity of water to neutralize acids. The
predominant chemical system present in natural waters is one where carbonates,
bicarbonates and hydroxides are present. The bicarbonate ion is usually
prevalent. However, the ratio of these ions is a function of pH, mineral
composition, temperature and ionic strength. Water may have a low alkalinity
rating but a relatively high pH or vice versa, so alkalinity alone is not of major
importance as a measure of water quality. Alkalinity is not considered detrimental
to humans but is generally associated with high pH values, hardness and excess
dissolved solids. High alkalinity waters may also have a distinctly flat, unpleasant
taste. Based on the testing, it is found that the alkalinity of the sample is 83 mg/L.
As per the provisional code, alkalinity should not exceed 200 mg/L for potable
water. For the fresh water alkalinity ranges between 20 100 mg/L. Alkalinity of
tested sample is within the limits specified in the standards. Hence the water
sample is fit for drinking.
5.10 EVALUATION
1. Alkalinity of water is an indication of
a) Base neutralizing capacity
b) Acid neutralizing capacity
c) Quantity of base present
d) Quality of base present
2. Mixed indicator is a combination of
a) Bromcresol Blue and Methyl Orange
b) Bromcresol Green and Methyl Red
c) Bromcresol Blue and Methyl Red
d) Bromcresol Green and Methyl Orange

3. Alkalinity is present due to all except _____.
a) Bromates
b) Phosphates
c) Silicates
d) Chlorides

4. Alkalinity is not caused by
a) Carbonates ions
b) Bicarbonates ions
c) Hydroxyl ions
d) Chloride ions
5. The phenolphthalein alkalinity is present then the pH of that water will be More
than
a) 8.3
b) 9.3
c) 7.3
d) 6.3

6. Alkalinity of natural water is mainly due to the presence of _______.

a) Bicarbonates
b) Bromates
c) Phosphates
d) Silicates

7. The bicarbonate equivalence point normally occur at pH
a) 2.5
b) 3.5
c) 4.5
d) 5.5

8. What is ppm?
a) Parts per meter square
b) Parts per meter
c) Parts per million
d) Parts per millimeter

9. The normality of the acid used in the titration is ___.

a) 0.2 N
b) 0.02 N
c) 0.002 N
d) 2.0 N

10. A standard solution is a
a) Solution of accurately known strength
b) Solution of accurately known pH
c) Coloured solution
d) Colourless solution

KEY TO ITEMS:
1) b
2) b
3) d
4) d
5) a
6) a
7) c
8) c
9) b
10) a

6.0 EXPERIMENT ON DETERMINATION OF ACIDITY OF
WATER
PREAMBLE:
How to determine acidity in Water and Wastewater.
procedure stated in
th
6.1 AIM
To determine the acidity of given water sample sample with the stipulations as
per IS: 3025 (Part 22) - Reaffirmed 2003.
6.2 INTRODUCTION
Acidity is a measure of the capacity of water to neutralise bases. Acidity is the
sum of all titrable acid present in the water sample. Strong mineral acids, weak
acids such as carbonic acid, acetic acid present in the water sample contributes
to acidity of the water. Usually dissolved carbon dioxide (CO
2
) is the major acidic
component present in the unpolluted surface waters.
The volume of standard alkali required to titrate a specific volume of the sample
to pH 8.3 is called phenolphthalein acidity (Total Acidity).
The volume of standard alkali required to titrate a specific volume of the water
sample (wastewater and highly polluted water) to pH 3.7 is called methyl orange
acidity (Mineral Acidity).
Acidity interferes in the treatment of water. Carbon dioxide is of important
considerations in determining whether removal by aeration or simple
neutralisation with lime /lime soda ash or NaOH will be chosen as the water
treatment method.
The size of the equipment, chemical requirements, storage spaces and cost of
the treatment all depends on the carbon dioxide present.
Aquatic life is affected by high water acidity. The organisms present are prone to
death with low pH of water.
High acidity water is not used for construction purposes. Especially in reinforced
concrete construction due to the corrosive nature of high acidity water.
Water containing mineral acidity is not fit for drinking purposes.
Industrial wastewaters containing high mineral acidity is must be neutralized
before they are subjected to biological treatment or direct discharge to water
sources.
6.3 PRINCIPLE
Hydrogen ions present in a sample as a result of dissociation or hydrolysis of
solutes reacts with additions of standard alkali (NaOH). Acidity thus depends on
end point of the indicator used.
The colour change of phenolphthalein indicator is close to pH 8.3 at 25C
corresponds to stoichiometric neutralisation of carbonic acid to bicarbonate.
1. Burette with Burette stand
2. porcelain tile
3. 500 mL conical flask
4. Pipette with elongated tips
5. Pipette bulb
6. Conical flask
7. Measuring cylinders
8. Wash Bottle and Beakers
1. Sodium Hydroxide
2. Phenolphthalein
3. Methyl Orange
4. Ethyl alcohol
5. Distilled Water

Preservation of sample is not practical. Because biological activity will
continue after a sample has been taken, changes may occur during
handling and storage.
To reduce the change in samples, keep all samples at 4 C. Do not allow
samples to Freeze.
Do not open sample bottle before analysis.
6.5.1 PRECAUTIONS
Colored and turbid samples may interfere in end point. Those samples
may be analyzed electrometrically, using pH meter.
Do not keep the indicator solution open since it contains the alcohol which
tense to evaporate. The mixed indicator solution is containing die in it.
Care should be taken so that it is not spill to your skin. If it spills on your
skin the scare will remain for at least 2 to 3 days.
Presence of residual chlorine may interfere in the colour response, which
can be nullified by addition of small amount of sodium thiosulphate or
destroy it with ultraviolet radiation.
Presence of iron and aluminum sulphate may interfere in the colour
response while titrating in room temperature, which can be nullified by
titrating the sample at boiling temperature.
Dissolved gases contributing to acidity such as CO
2
, H
2
S may interfere in
the titration, hence avoid vigorous shaking.
Samples suspected to have hydrolysable metal ions or reduced forms of
polyvalent cations need hydrogen per oxide treatment.
6.6 PROCEDURE
Sodium Hydroxide (0.02 N)
Take 1000 mL standard measuring flask and fill 3/4
th
of it with distilled
water.
Accurately measure 20 mL of 1N sulphuric acid solution using a pipette
and transfer to 1000 mL standard flask containing the distilled water. Make
up to 1000 mL using distilled water.
Phenolphthalein Indicator
Weigh accurately 1 g of phenolphthalein and dissolve it in 95% ethyl
alcohol.
Transfer it to the 100 mL standard flask and make up to 100 mL using
95% ethyl alcohol.
Methyl Orange Indicator
Weigh accurately 1 g of methyl and dissolve it in distilled water.
Transfer it to the 100 mL standard flask and make up to 100 mL using
distilled water.
Rinse the burette with 0.02N sodium hydroxide and then discard the
solution.
Fill the burette with 0.02N sodium hydroxide and adjust the burette.
Fix the burette to the stand.
A sample size is chosen as the titre value does not exceed 20mL of the
titrant. For highly concentrated samples, dilute the sample. Usually, take
100 mL of a given sample in a conical flask using pipette.
Add few drops of methyl orange indicator in the conical flask.
The colour changes to orange. Now titrate the sample against the 0.02N
sodium hydroxide solution until the orange colour faints.
Note down the volume (V
1
) consumed for titration _____mL. This
volume is used for calculating the mineral acidity.
To the same solution in the conical flask add few drops of phenolphthalein
indicator.
Continue the titration, until the colour changes to faint pink colour.
Note down the total volume (V
2
) consumed for titration _____ mL.
This volume is used for calculating the total acidity.
6.7 CALCULATION
6.7.1 TABLE
Table -1 Mineral Acidi-
ty:
Burette Solution: Sodium Hydroxide
Pipette Solution: Sample
Indicator: Methyl Orange
End Point: Faint of Orange Color
For the calculation of Mineral Acidity:
The Sodium Hydroxide is taken in the burette.
For the First titration the volume of water sample taken is 100 mL.The
initial reading is _____, the final reading is ______ mL.
The volume of NaOH consumed to get the end point is _____mL.
For Second titration the volume of water sample taken is 100 mL.The
initial reading is ______, the final reading is _______mL.
The volume of NaOH consumed to get the end point is ______ mL.
For third titration the volume of water sample taken is 100mL.The initial
reading is _____, the final reading is ______ mL.
The volume of NaOH (V
1
) consumed to get the end point is _____ mL.
For second and third titration the burette reading is same so we have
achieved concordant values. We can go for the calculations.
Sl.No.
Volume of
Sample (mL)
NaOH (mL)
Initial Final
1.
2.
3.
Table -2 Total Acidity:
Burette Solution: Sodium Hydroxide
Indicator: Phenolphthalein
End Point: Faint Pink Color
For the calculation of Total Acidity:
The Sodium Hydroxide is taken in the burette.
For the First titration the volume of water sample taken is 100 mL.The
initial reading is ______, the final reading is ______ mL.
For Second titration the volume of water sample taken is 100 mL.The
initial reading is ______, the final reading is ______ mL.
For third titration the volume of water sample taken is 100 mL. The initial
reading is ______, the final reading is _____ mL.
The volume of NaOH (V
2
) consumed to get the end point is ______ mL.
For second and third titration the burette reading is same so we have
achieved concordant values. We can go for the calculations.
Sl.No.
Volume of
Sample (mL)
NaOH (mL)
Initial Final
1.
2.
3.
6.7.2 DATA SHEET
DETERMINATION OF ACIDITY
DATA SHEET
Date Tested :
Tested By :
Project Name :
Sample Number :
Sample Location :
Table - 1 for Mineral Acidity:
Table - 2 for Total Acidity:
Model Calculation:
Volume of NaOH for Mineral Acidity (V1) =
Volume of NaOH for Total Acidity (V2) =
Volume of Sample =
Mineral Acidity =
Sl.No.
Volume of NaOH (V1) * N * 50 * 1000
Volume of
Sample (mL)
NaOH (mL)
Initial Final
4. 100
5. 100
6. 100
Sl.No.
Volume of
Sample (mL)
NaOH (mL)
Initial Final
1. 100
2. 100
3. 100
To convert the sample size from mL to L, multiply the result by 1,000 mL/L
Mineral Acidity as CaCO3 equivalent (mg/L) =
=
Total Acidity = Volume of NaOH (V2) * N * 50 * 1000
Total Acidity as CaCO3 equivalent (mg/L) =
=
The Mineral Acidity as CaCO
3
equivalent is =______mg/L
The Total Acidity as CaCO
3
equivalent is =_______ mg/L.
6.9 INFERENCE
Acidity 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. Acidity may contribute to corrosiveness and influence chemical
reaction rates, chemical speciation and biological process. The measurement
also reflects a change in the quality of the source water. Strong mineral acids,
weak acids such as carbonic acid, acetic acid and hydrolyzing salts such as iron
or aluminum sulphates may contribute to the measured acidity
.
6.10 EVALUATION
1. Acidity is _____.
a) Base neutralizing capacity
b) Acid neutralizing capacity
c) Quantity of acid present
d) Quality of acid present
2. An Indicator is a substance that facilitate colour change at the end point.
a) True
b) False

3. The indicators used in the titration are
a) Methyl orange and phenolphthalein
b) Methyl red and phenolphthalein
c) Methyl orange and Methyl red
d) Bromocresol green and Methyl red
4. To prepare 100 mL of 0.02 N of NaOH from 1 N NaOH, dilute ________
of
NaOH.
a) 20 mL
b) 2 mL
c) 0.2 mL
d) 0.02 mL

5. The major acidic component of surface water is

a) Dissolved oxygen
b) Dissolved carbon di oxide
c) Dissolved sulphur di oxide
d) Dissolved nitrous oxide

6. The end point determination in titration will be based on the __________.

a) Temperature
b) Hardness
c) Residual Chlorine
d) Conductivity

7. The methyl orange acidity is at pH ______.

a) 3.7
b) 3.9
c) 4.5
d) 4.7

8. The phenolphthalein acidity is at pH is 8.3

a) 8.3
b) 9.3
c) 4.3
d) 7.3

9. For dilution purposes, ____________ type of distilled water is used.

a) Organic free
b) CO
2
free
c) O
2
free
d) Ordinary

10. Acidity can be electrometrically measured by_______________

a) pH meter
b) Conductivity meter
c) Turbidity meter
d) Spectrometer

KEY TO ITEMS:

1) a
2) True
3) a
4) b
5) b
6) c
7) a
8) a

9) b
10) a

7.0 EXPERIMENT ON DETERMINATION OF CHLORIDES
PREAMBLE:
How to determine chlorides in Water and Wastewater.
procedure stated in
th
Edition. Method 4500 - Cl
-
- B.
USEPA, Method 9253.
7.1 AIM
To determine the chlorides of given water sample with the stipulations as per
7.2 INTRODUCTION
Chlorides are widely distributed as salts of calcium, sodium and potassium in
water and wastewater. In potable water, the salty taste produced by chloride
concentrations is variable and dependent on the chemical composition of water.
The major taste producing salts in water are sodium chloride and calcium
chloride. The salty taste is due to chloride anions and associated cations in water.
In some water which is having only 250 mg /L of chloride may have a detectable
salty taste if the cat-ion present in the water is sodium. On the other hand, a
typical salty taste may be absent even if the water is having very high chloride
concentration for example 1000 mg /L.
This is because the predominant cation present in the water is not sodium but
either calcium or magnesium may be present.
7.2.1 Environmental Significance
Chlorides associated with sodium (Sodium Chloride) exert salty taste
when its concentration is more than 250 mg/L. These impact a salty taste
to water. Chlorides are generally limited to 250 mg/L in water supplies
intended for public water supply.
In many areas of the world where water supplies are scarce, sources
containing as much as 2000 mg/L are used for domestic purposes without
the development of adverse effect, once the human system becomes
adapted to the water.
It can also corrode concrete. Magnesium chloride in water generates
hydrochloric acid after heating which is also highly corrosive and creates
problem in boilers.
Chloride determinations in natural waters are useful in the selection of
water supplies for human use.
Chloride determination is used to determine the type of desalting
apparatus to be used.
Chloride determination is used to control pumping of ground water from
locations where intrusion of seawater is a problem.
Chlorides interfere in the determination of chemical oxygen demand
(COD).
7.3 PRINCIPLE
The amount of chloride present in water can be easily determined by titrating the
given water sample with silver nitrate solution.
The silver nitrate reacts with chloride ion according to1 mole of AgNO
3
reacts
with 1 mole of chloride. The titrant concentration is generally 0.02 M.
Silver chloride is precipitated quantitatively, before red silver chromate is formed.
The end of titration is indicated by formation of red silver chromate from excess
silver nitrate.
The results are expressed in mg/L of chloride (Cl
-
with a molecular weight of
35.453 g/mol).
1. Burette with Burette stand and porcelain tile
3. Conical flask (Erlenmeyer Flask)
4. Standard flask
5. Beaker
6. Wash bottle

1. Silver nitrate
2. Phenolphthalein Indicator
3. Sodium chloride
4. Potassium chromate

If Analysis is to be carried out with in two hours of collection, cool storage is not
necessary. If analysis can not be started with in the two hours of sample
collection to reduce the change in sample, keep all samples at 4
0
C.
Do not allow samples to freeze. Do not open sample bottle before analysis.
Begin analysis within six hours of sample collection
7.5.1 PRECAUTIONS
AgNO
3
should be stored in a brown amber bottle and should not be
exposed to sunlight.
While handling AgNO
3
, care should be taken so that it is not spilled on
your skin.
If it spills on your skin, the scar will remain at least for ten to fifteen days.
7.6 PROCEDURE
Standard Sodium Chloride Solution
Switch on the Electronic balance, keep the weighing pan, and set the
reading to zero.
Weigh 1.648g of Sodium chloride
Transfer the contents to the beaker containing distilled water. Using
glass rod, dissolve the contents thoroughly.
Transfer the contents in the beaker to a 100 mL standard flask; fill
distilled water up to 100 mL mark.
Transfer it to 100mL standard flask using funnel
Standard Silver Nitrate (0.0282 N)
Weigh 4.791g of Silver nitrate and transfer it to the beaker with distilled
water.
Transfer the contents in the beaker to a 100 mL standard flask, fill
distilled water up to 100 mL mark.
Standardize it against 0.0282 N NaCl solution. Store it in an amber
bottle.
Potassium Chromate Indicator
Weigh 25 g of Potassium Chromate. Transfer it to the beaker contains
distilled water. Add few drops of Silver Nitrate solution until slight red
precipitate is formed.
Allow it to stand for 12 hours. After 12 hours filter the solution using
filter paper and dilute the filtrate to 1000 mL using distilled water.
Before starting the titration rinse the burette with silver nitrate solution.
Fill the burette with silver nitrate solution of 0.0282 N. Adjust to zero
and fix the burette in stand.
Take 20 mL of the sample in a clean 250mL conical flask
Add 1 mL of Potassium Chromate indicator to get light yellow color
Titrate the sample against silver nitrate solution until the color changes
from yellow to brick red. i.e., the end point.
Note the volume of Silver nitrate added (A).
The value of titration is _____ mL.
Repeat the procedure for concordant values.
Blank Titration
Take 20 mL of the distilled water in a clean 250mL conical flask
Add 1 mL of Potassium Chromate indicator to get light yellow color
Titrate the sample against silver nitrate solution until the color changes
from yellow to brick red. i.e., the end point.
Note the volume of silver nitrate added for distilled water (B).
The value of titration is _______ mL
7.7 CALCULATION
7.7.1 TABLE
Sample
No
Volume of
Sample (mL)
Volume of AgNO
3

(mL)
Initial Final
1.
2.
Blank (B)
Burette solution: Silver Nitrate
Pipette solution: Sample
Indicator: Potassium chromate
End point: Appearance of Brick red color.
The volume of water sample taken is 20 mL.
The silver nitrate is taken in the Burette.
For the first titration, the initial reading is ______mL. The final reading
is ______ mL.
The volume of silver nitrate consumed to get the end point is ____ mL.
For the second titration, the initial reading is ______ mL. The final
reading is
______mL.
The volume of water sample taken is 20 mL.
The silver nitrate is taken in the Burette.
For the first titration, the initial reading is ______ mL. The final read-
ing is ______ mL.
The volume of silver nitrate consumed to get the end point is ____ mL.
For the second titration, the initial reading is ______ mL. The final
reading is ______ mL.
The volume of silver nitrate consumed to get the end point is ___ mL.
For the first and second titration, the burette reading is same so we
have achieved concordant value. We can go for the calculations
For the blank titration the end point is attained within the few drops of
silver nitrate
So the burette reading is _____ mL.
Total amount of Chlorides mg/L of the given water sample is equal to
Volume of AgNO
3
used for sample minus AgNO
3
used for blank
multiplied by Normality multiplied by 35.45 multiplied by 1000 divided
by Volume of sample taken
Here the volume of silver nitrate used for sample is
--------mL and for blank is ______mL
Normality is _______ N
volume of sample taken is 20 mL Substituting the values in the formula
and calculating we get the value ________mg/L
7.7.2 DATA SHEET
DETERMINATION OF CHLORIDES
DATA SHEET
Date Tested :
Tested By :
Project Name :
Sample Number :
Sample Location :
Volume of Silver Nitrate for sample (Vs) =
Volume of Silver Nitrate for Blank (VB) =
Normality of EDTA =
Volume of Sample =
Equivalent weight of Chlorine =
Chlorides mg/ L = (V
s
V
B
) * Normality * 35.45 * 1000
Chlorides mg/ L =
=
Sl.No.
Volume of
Sample (mL)
EDTA (mL)
Initial Final
1. 20
2. 20
Blank (B) 20
The amount of chloride present in the given water sample is =______ mg/L.
7.9 INFERENCE
The high concentrations of chloride ions mostly results in an unpleasant salty
taste of water and it also aides the corrosion of plumbing system. Very high
chloride content of water may also produce laxative effect. An upper limit of 250
mg/L has been set for the chloride ions. An increase in the normal chloride
content of your water may indicate possible pollution from human sewage,
animal manure or industrial wastes. As all aware the sea water is full of sodium
chloride, the chloride levels will be much higher compared to the fresh water
sources.
7.10 EVALUATION
1. The limit of chlorides in drinking water as per IS code is
a) 200 ppm
b) 225 ppm
c) 250 ppm
d) 500 ppm
2. Silver nitrate is stored in a brown bottle
a) to avoid decomposition by sun light
b) because it is dark in colour
c) because the solution is colourless
d) to avoid heat

3. The colour of Silver Chromate is
a) Milky White
b) pale Yellow
c) Colourless
d) Brick Red

4. When both hardness and chloride content are very high above 500 mg/L, then
the water will be

a) Non salty in nature
b) Fit for drinking
c) Salty in nature
d) Soft water

5. Presence of chloride can corrode________.

a) GI pipes
b) Rubber tubes
c) PVC pipes
d) Glass pipes

6. The chloride concentration in sewage is

a) More concentrated than the municipal water supplied
b) Equal concentration to the municipal water supplied
c) Less concentrated than the municipal water supplied
d) Only in trace

7. Chloride consumed by human beings

a) Pass through the fecal matter as it is
b) Gets changed into other forms
c) Gets disappeared in the body
d) Stored in bones

8. Chloride gives salty taste to water particularly when present as ___.

a) Sodium chloride
b) Magnesium chloride
c) Potassium chloride
d) Zinc chloride

9. The point at which a clear visual change is observed after the reaction
between titrant and titrates is called

a) End point
b) Equivalence point
c) Equal point
d) Double equivalence point

10. Most common ion in the water is

a) Fluoride
b) Nitrate
c) Chloride
d) Sulphate

KEY TO ITEMS:

1) c
2) a
3) d
4) c
5) a
6) a
7) a
8) a
9) a
10) c
8.0 EXPERIMENT ON DETERMINATION OF TOTAL SOLIDS IN
WATER
PREAMBLE:
How to determine total solids in Water and Wastewater.
stated in
th
Edition.
Method 2540 B.
Method 160.3.
8.1 AIM
To determine the total solids in the given water sample. Test procedure is in
accordance to IS: 3025 (Part 15) - Reaffirmed 2003.
8.2 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.
Solids can be classified as either filterable or non filterable. Filterable solids may
either be settleable or non settleable. Solids can also be classified as organic or
inorganic.
Total Solids is the term applied to the material residue left in the vessel after
evaporation of a sample and its subsequent drying in an oven at a defined
temperature.
Measurement of Solids can be made in different water samples (industrial, domestic
and drinking water) and it is defined as residue upon evaporation of free water.
Thus, Total solids are nothing but summation of total dissolved solids and
total suspended solids.
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.
Total solids also affect water clarity. Higher solids decrease the passage of light
through water, thereby slowing more rapidly and hold more heat; this, in turn, might
adversely photosynthesis by aquatic plants. Water will heat up affect aquatic life that
has adapted to a lower temperature regime.
As with turbidity, concentrations often increase sharply during rainfall, especially in
developed watersheds. They can also rise sharply during dry weather if earth-
disturbing activities are occurring in or near the stream without erosion control
practices in place.
Regular monitoring of total solids can help detect trends that might indicate
increasing erosion in developing watersheds.
Total solids are related closely to stream flow and velocity and should be correlated
with these factors. Any change in total solids over time should be measured at the
same site at the same flow.
In the case of water:
Water with total solids generally is of inferior palatability and may induce an
unfavorable physiological reaction. It may be esthetically unsatisfactory for purposes
such as bathing.
Total solids will be higher in highly mineralized waters, which result in unsuitability
for many industrial applications.
It indicates effectiveness of sedimentation process and it affects effectiveness of
disinfection process in killing microorganisms.
It is used to assess the suitability of potential supply of water for various uses. In the
case of water softening, amount of total solids determine the type of softening
procedure.
Corrosion control is frequently accomplished by the production of stabilized waters
through pH adjustment. The pH stabilization depends to some extent upon the total
solids present as well as alkalinity and temperature.
In the case of waste water:
Solids analyses are important in the control of biological and physical wastewater
treatment processes and for assessing compliance with regulatory agency
wastewater effluent limitations
Although the waste water or sewage normally contains 99.9 percent of water and
only 0.1 percent of solids, but it is the solids that have the nuisance value.
The amount of solids in wastewater is frequently used to describe the strength of the
water. The more solids present in a particular wastewater, the stronger that
wastewater will be. The environmental impacts of solids in all forms have detrimental
effects on quality since they cause putrefaction problems.
If the solids in wastewater are mostly organic, the impact on a treatment plant is
greater than if the solids are mostly inorganic.
8.3 PRINCIPLE
The sample is evaporated in a weighed dish on a steam bath and is dried to a
constant mass in an oven either at 103-105C or 179-181C.
Total solids/residue is calculated from increase in mass.
1. Crucible
2. Oven
3. Desiccators
4. Analytical Balance
5. Dish Tongs
6. Magnetic Stirrer
7. Wash Bottle

Preservation of sample is not practical. Because biological activity will continue after
a sample has been taken, changes may occur during handling and storage.
To reduce this change in samples taken for solids determinations, keep all samples
at 4
0
C.
Do not allow samples to freeze.
8.5.1 PRECAUTIONS
Water or Wastewater samples which contain high concentrations of calcium,
chloride, magnesium or sulphate can rapidly absorb moisture from the air.
Such samples may need to be dried for a longer period of time, cooled under
proper desiccation and weighed rapidly in order to achieve a reasonable
constant weight.
Non-representative particulates such as leaves, sticks, fish and lumps of fecal
matter should be excluded from the sample if it is determined that their
inclusion is not desired in the final result.
Floating oil and grease, if present, should be included in the sample and
dispersed by a blender device before sub-sampling.
Volume of sample should be adjusted to have residue left after drying as 100
to 200mg. It is mainly to prevent large amount of residue in entrapping water
during evaporation.
Highly mineralized water containing significant concentration of calcium,
magnesium, chloride, and/or sulphate may be hygroscopic. Hence prolonged
drying, desiccation and rapid weighing.
Volume of sample should be adjusted to have residue left after drying as 100
to 200mg. It is mainly to prevent large amount of residue in entrapping water
during evaporation.
8.6 PROCEDURE
To measure total solids, take a clean porcelain dish which has been washed
and dried in a hot air oven at 105C for one hour.
Now weigh the empty evaporating dish in analytical balance. Lets denote the
weight measured as (W1).
Now we should have to decide what should be the volume of sample to be
taken for analysis.
Volume may be estimated either from values of specific conductance or
general thumb rule.
In general, select a sample volume that will yield residue between 2.5 and
200 mg after drying.
Switch on the oven and allowed to reach 105C. Check and regulate oven
and furnace temperatures frequently to maintain the desired temperature
range.
Place it in the hot air oven and care should be taken to prevent splattering of
sample during evaporation or boiling.
Dry the sample to get constant mass. Drying for long duration usually 1 to 2
hours is done to eliminate necessity of checking for constant mass.
Cool the container in a desiccator. Desiccators are designed to provide an
environment of standard dryness. This is maintained by the desiccant found
inside. Don't leave the lid off for prolonged periods or the desiccant will soon
be exhausted.
Keep desiccator cover greased with the appropriate type of lubricant in order
to seal the desiccator and prevent moisture from entering the desiccator as
the test glassware cools.
We should weigh the dish as soon as it has cooled to avoid absorption of
moisture due to its hygroscopic nature.
Samples need to be measured accurately, weighed carefully, and dried and
cooled completely.
Note the weight with residue as (W
2
).
8.7 CALCULATION
Initial weight of the Crucible (W
1
) =.. g
Final weight of the Crucible +sample (W
2
) =.. g
Weight of residue (W) =W
2
- W
1
g
Amount of total solids present in the sample =
v
w 1000 * 1000
W =weight of total residue in (mg). (Therefore multiply W with 1000)
V =Volume of the sample (mL)(To convert mL to L)
=..mg/L
The readings are required to be tabulated.
14.7.1 TABLE
8.7.2 DATA SHEET
DETERMINATION OF TOTAL SOLIDS
DATA SHEET
Date Tested :
Tested By :
Project Name :
Sample Number :
Sample Location :
W1 =
W2 =
V =
=
=
1000)
W (mg) =
= 41.6mg
Description
Total Solids (mg/L)
V = Volume of the sample (mL) (To convert mL to L, multiply by 1000)
=41.6 mg/75 mL = 0.555 mg/mL
= 0.555 mg/mL x 1,000 mL/L
= 555 mg/L
Weight
(g)
Initial Weight of the Crucible (g) W
1
Final Weight of the Crucible + sample (g) W
2
Total Solids (mg/L) TS
8.8 Interpretation of Results
In the given sample, a total solid is equivalent to _______ mg/L.
8.9 Inference
Total solids are nothing but summation of Total Dissolved Solids and Total
Suspended Solids. Regular monitoring of total solids can help detect trends that
might indicate increasing erosion in developing watersheds. Total solids also affect
water clarity. Total Solids may indicate the presence of agricultural activities,
dredging, or mining upstream from your sample site.
8.10 Evaluation
1. After Evaporation, the evaporating dishes needs to be
.
a) weighed immediately
b) kept in air for cooling to room temperature
c) cooled to room temperature in a dessicator
d) cooled to a temperature less that 25C
2. Total Solids are referred to materials left after evaporation.
a) True
b) False
3. A sample was stored @ 4 C for 4 days. During the analysis, the temperature of
the sample should be
a) maintained at 4 C
b) brought to room temperature
c) below room temperature
d) brought above room temperature by adding boiled distilled water
4. The determination of total solids in wastewater gives an idea about
a) the foulness of the sewage
b) pH of the sewage
c) temperature of the sewage
d) colour of the sewage
5. The evaporating dishes needs to be cleaned and dried at _______ to remove the
existing organic content.

a) 100 C
b) 250 C
c) 450 C
d) 550 C

6. Sewage contains about 99% of _____.

a) water
b) solids
c) clay
d) microbes

7. Interference in the determination of total solids is due to ______.

a) Oil and Greese
b) Large water sample
c) Dissolved salts
d) Suspended salts

8. For analysis of total solids the sample used should be
a) homogenous sample
b) supernatant of the sample
c) settled sample
d) clear sample

9. The sewage contain
a) suspended and dissolved solids.
b) no solids
c) only dissolved solids
d) only suspended solids

10. The major dissolved substances in natural water are comprised of
a) iron, manganese, silica and nitrate
b) calcium, magnesium, sodium, bicarbonate, sulfate and chloride
c) all anions
d) all cations

KEY TO ITEMS:
1) c
2) True
3) b
4) a
5) d
6) a
7) a
8) a
9) a
10) b
9.0 EXPERIMENT ON DETERMINATION OF TOTAL ORGANIC AND
INORGANIC SOLIDS IN WATER
PREAMBLE:
How to determine total organic and inorganic solids in Water and Wastewater.
In addition to our Indian Standard, we also discuss in brief regarding the procedure stated
in
th
Edition. Method
2540 E.
(2) Methods for Chemical Analysis of Water and Wastes, EPA-600/4-79-020, USEPA, Method
160.4.
9.1 AIM
To determine total organic and inorganic solids in the given water sample with the
stipulations as per IS: 3025 (Part 18) - Reaffirmed 2002.
9.2 INTRODUCTON
The term total volatile solids refer to materials that are completely volatilised from water at
higher temperature (550C). These solids are often referred to the organic content of the
water. The term total fixed solids can be referred to materials which are not volatilised
from water at higher temperature (550C). These solids are often referred to the inorganic
content of the water.
The water which consists of high volatile solids is not suitable for drinking purpose
and indicates that the water may have been polluted by domestic wastes or other
organic wastes.
Volatile solids test is normally applied to sludges. It is indispensable in the design
and operation of sludge digest, vacuum filter and incineration plants.

Before the development of the COD test, it is used to find out the strength of
industrial and domestic wastewater. It is helpful in assessing the amount
biologically inert organic matter, such as lignin in case of wood pulping waste
liquours.
The determination of volatile and fixed components in the residue is useful in the
control of waste water plant operation because it offers an approximate amount of
organic matter present in the solid fraction of wastewater.
9.3 PRINCIPLE
The sample is evaporated in a weighed dish on a steam bath and is dried to a constant
mass in an oven at 103-105C. The residue obtained is ignited to constant weight at 550(C. The
remaining solids represent the total fixed solids and the weight lost during the ignition represents
the total volatile solids.
1. Evaporating Dish
2. Water Bath (Steam Bath)
3. Oven
4. Desiccators
5. Weighing balance
6. Dish Tongs
7. Magnetic Stirrer
8. Wash Bottle

Preservation of sample is not practical. Because biological activity will continue after a
sample has been taken, changes may occur during handling and storage.
To reduce this change in samples taken for solids determinations, keep all samples at
4
C. Do not allow samples to freeze.

9.5.1 PRECAUTIONS
Negative errors in volatile solids may be produced by loss of volatile matter during
drying in the oven.
In the presence of high concentration fixed solids, the determination of low
concentration of volatile solids may be subject to considerable error. In those
cases, the measure of volatile components by some other method like total organic
carbon is advisable.
Floating oil and grease, if present, should be included in the sample and dispersed
by a blender device before sub-sampling.
Volume of sample should be adjusted to have residue left after drying as 100 to
200mg. It is mainly to prevent large amount of residue in entrapping water during
evaporation.
9.6 PROCEDURE
To measure total volatile solids and fixed solids, take a clean silica crucible which
has been washed and dried in a hot air oven at 105C for one hour and ignited at
550C to remove all organic materials present in it.
Now weigh the empty silica crucible in analytical balance. Lets denote the weight
measured as W1 =________g
Switch on the oven and allowed to reach 105C. Check and regulate oven and
furnace temperatures frequently to maintain the desired temperature range.
Place the silica crucible in the hot air oven and care should be taken to prevent
splattering of sample during evaporation or boiling.
Dry the sample to get constant mass. Drying for long duration is done to eliminate
necessity of checking for constant mass.
Cool the container in a desiccator. Desiccators are designed to provide an
environment of standard dryness. This is maintained by the desiccant found inside.
Don't leave the lid off for prolonged periods or the desiccant will soon be
exhausted.
We should weigh the dish as soon as it has cooled to avoid absorption of moisture
due to its hygroscopic nature.
Samples need to be measured accurately, weighed carefully, and dried and cooled
completely.
2
= ________ g
Switch on the furnace and allow it to reach 550C. Check and regulate the furnace
temperatures frequently to maintain the desired temperature range.
Place the silica crucible in the furnace and care should be taken while keep the
crucible inside the furnace since it will be too hot.
Allow it to ignite for 20 minutes to get constant mass.
As above, cool the silica crucible in a desiccator to room temperature.
Weigh the dish as soon as it has cooled to avoid absorption of moisture due to its
hygroscopic nature.
3
= _________ g
9.7 CALCULATION
Total Volatile Solids
Initial weight of the evaporating dish +sample (W
1
) =.. g
Final weight of the evaporating dish +sample after drying at 105C (W
2
) =.. g
3
) =.. g
Weight of volatile substance (W) =W
2
W
3
g
Amount of total solids present in the sample =
V =Volume of the sample (mL) (To convert mL to L)
=..mg/L
Total Fixed Solids
Initial weight of the evaporating dish (W
1
) =.. g
2
) =.. g
3
) =.. g
Weight of non volatile substance (W) =W
3
W
1
g
Amount of total fixed solids present in the sample =
V =Volume of the sample (mL) (To convert mL to L)
=..mg/L
9.7.1 TABLE Total
Volatile Solids
The Weight of the clean silica crucible (g) W
1
=
The Weight of the clean silica crucible and the residue (g) W
2
=
The Weight of the residue (g) W=
Weight of the clean silica crucible (g) W
1

Weight of the silica crucible and the residue (g) W
2

Weight of residue (g) W
Weight of the silica crucible and the ash (g) W
3

Weight of ash (g) W
Total Volatile Solids (mg/L) TVS
The Weight of the silica crucible and the ash (g) W
3
= ______ g
Weight of the ash (g) W

= ________
The volume of the sample (mL) V = 100 mL
Total Fixed Solids
The Weight of the clean silica crucible (g) W
1
=
The Weight of the silica crucible and the residue (g) W
2
=
Weight of the residue (g) W=
3
=
Weight of the ash (g) W
a
=
Volume of the sample (mL) V =
1

2

3

Weight of ash (g) W
Total Fixed Solids (mg/L) TFS
9.7.2 DATA SHEET
DETERMINATION OF TOTAL VOLATILE SOLIDS
DATA SHEET
Date Tested :
Tested By :
Project Name :
Sample Number :
Sample Location :
Speicmen Calculation:
W2 =
W3 =
V =
Weight of residue (g) W = W2 W3
=

Weight of residue in mg (To convert W (g) to W (mg), multiply W (g) with 1000)
W (mg) =

Description
Total Volatile Solids (mg/L)
1000)
Weight
(g)
1
2
3
Weight of ash (g) W
Total Volatile Solids (mg/L) TVS
DETERMINATION OF TOTAL FIXED SOLIDS
DATA SHEET
Date Tested :
Tested By :
Project Name :
Sample Number :
Sample Location :
W1 =
W3 =
V =

1000)
W (mg) =
Description
Total Fixed Solids (mg/L)
1000)

Weight
(g)
1
2
3
Weight of ash (g) W
Total Fixed Solids (mg/L) TFS
In the given sample, total volatile solids is equivalent to _______ mg/L and total fixed
solids is ______mg/L.
9.9 INFERENCE
In domestic wastewater, solids are about 50 percent organic, which in turn contaminates
the ground and fresh water. These solids are generally from vegetable, dead animal
matter, and also include synthetic organic compounds. They can be ignited or
burned. Since the organic fraction can be driven off at high temperatures, they are called
volatile solids. Inorganic solids are frequently called mineral substances and include sand,
gravel and silt as well as the mineral salts in the water supply which produce the
hardness and mineral content of the water. Mostly, they are non-combustible. They are
called non volatile solids.
9.10 EVALUATION
1. The Total Volatile Solids determination is very important in the control of
a) Water treatment plant
b) Sewage treatment plant
c) Desalination plant
d) Effluent treatment plant
2. The crucible with sample, should be placed in the muffle furnace for atleast _______.
a) one hour
b) two hours
c) 20 minutes
d) 10 minutes
3. The Total Fixed Solids is the measure of
a) all the solids present
b) inorganic solids present
c) the salt content
d) organic solids
4. The method used for the determination of solids is _____.

a) volumetric method
b) gravimetric method
c) instrumentation method
d) visual method

5. The crucible after ignition should be cooled in a desiccator
a) because it is hot
b) to avoid moisture absorption
c) to cool
d) to incubate

6. Putrescible solid means
a) pure solids
b) dissolved solids
c) solids with high BOD
d) suspended solids

7. The solid organic matter (sludge) digested by Aerobic treatment.
a) True
b) False

8. The determination of total volatile solids is interfered by

a) Loss of volatile solids during the drying process
b) Large volatile solids water sample
c) Dissolved salts
d) Suspended salts

9. While placing the crucible in muffle furnace it is advisable to wear gloves made of

a) Leather
b) Rubber
c) Resin
d) Polythene

10. The Total Volatile Solids is the measure of

a) all the solids present
b) organic solids present
c) the salt content
d) inorganic salts present

KEY TO ITEMS:
1) b
2) c
3) b
4) b
5) b
6) c
7) False
8) a
9) a
10) b
10.0 EXPERIMENT ON DETERMINATION OF DISSOLVED OXYGEN
PREAMBLE:
How to determine dissolved oxygen in Water and Wastewater.
stated in
th
Edition. Method 4500-O G.
10.1 AIM
To determine dissolved oxygen (DO) in the given water sample with the
10.2 INTRODUCTION
Before performing this experiment, few questions may arise to the learners:
1. What is meant by Dissolved Oxygen (DO)? Is it oxygen in dissolved form?
2. Why we need to determine DO?
3. What are the methods available to determine DO?
4. Is it measured in natural water or wastewater?
5. Whether is it mandatory as per our codal provision to determine DO?
The term Dissolved Oxygen is used to describe the amount of oxygen dissolved in
a unit volume of water. Dissolved oxygen (DO) is essential for the maintenance of
healthy lakes and rivers. It is a measure of the ability of water to sustain aquatic
life.
The dissolved oxygen content of water is influenced by the source, raw water
temperature, treatment and chemical or biological processes taking place in the
distribution system.
The presence of oxygen in water is a good sign. Depletion of dissolved oxygen in
water supplies can encourage the microbial reduction of nitrate to nitrite and
sulfate to sulfide. It can also cause an increase in the concentration of ferrous iron
in solution, with subsequent discoloration at the tap when the water is aerated.
Hence, analysis of dissolved oxygen is an important step in water pollution control
and wastewater treatment process control. There are various methods available to
measure Dissolved Oxygen, which we will discuss in detail.

In a healthy body of water such as a lake, river, or stream, the dissolved oxygen is
about 8 parts per million. The minimum DO level of 4 to 5 mg/L or ppm is desirable
for survival of aquatic life.

Now imagine that a source of oxygen demanding wastes, such as feed lot, a paper
mill or a food processing plant, is built besides the river. The facility begins
operating and discharging wastes into the river.

This increases the BOD and affects the concentration of DO in the waters
downstream.

The wastes serve as the food for certain aerobic bacteria. as it moves
downstream, the conc. of bacteria increases. Because these bacteria remove
oxygen from water, their population increase causes a decline in the amount of
DO.

Beyond certain point, most of the wastes break down. The conc. of DO rises as the
river recovers oxygen from the atmosphere and aquatic plants.
Thus DO test is the basis for BOD test which is an important parameter to evaluate
organic pollution potential of a waste.

It is necessary for all aerobic biological wastewater treatment processes to control
the rate of aeration.

Drinking water should be rich in dissolved oxygen for good taste.
DO test is used to evaluate the pollution strength of domestic and industrial waste.
Higher values of DO may cause corrosion of Iron and Steel.
Algae growth in water may release oxygen during its photosynthesis and DO may
even shoot upto 30 mg/L.
Oxygen is poorly soluble in water. Its solubility is about 14.6 for pure water at 0C
under normal atmospheric pressure and it drops to 7 mg/l at 35C.
Higher temperature, biological impurities, Ammonia, Nitrates, ferrous iron,
chemicals such as hydrogen sulphide and organic matter reduce DO values.
Aerobic bacteria thrive when oxygen is available in plenty. Aerobic conditions do
prevail when sufficient DO is available within water. End products of aerobiosis are
stable and are not foul smelling.
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 for BOD test which is an important parameter to evaluate
organic pollution potential of a waste.
DO test is necessary for all aerobic biological wastewater treatment processes to
control the rate of aeration.

10.3 PRINCIPLE
Dissolved Oxygen can be measured either by titrimetric or electrometric method.
(1) Titrimetric Method
Titrimetric method is based on the oxidizing property of DO while the electrometric
method (using membrane electrodes) is based on the rate of diffusion of molecular
oxygen across a membrane. It is most accurate method to determine DO.

There are different titrimetric methods based on the nature of sample to be tested.
(a) Winkler Method
(b) Azide Modification
(c) Alum Flocculation Modification
(d) Permanganate Modification

However, in all the above the basic principle remains same.
Choice of the method depends upon the type of sample to be tested

Azide Modification:
In this method, interference caused by nitrate is removed effectively. Presence of
nitrate is most interference in biologically treated effluent and incubated BOD
samples.
Al um Flocculation Modification:
If the sample contains suspended solids (especially effluent samples), then this
method will be suitable.
Permanganate Modification:
If the sample contains iron (Fe2+) ions. Addition of 1mL of potassium fluoride and
azide solution can be adopted to suppress the interference due to (Fe3+).
This method is not useful when the sample contains sulphites, thiosulphates and
high BOD.
The Titrimetric principle:
Divalent Manganese salt in solution is precipitated by strong alkali to divalent
manganese hydroxide.

Addition of Potassium iodide or Potassium hydroxide is added to create a pinkish
brown precipitate.
In the alkaline solution, dissolved oxygen present in the sample rapidly oxidized to
form trivalent or higher valency hydroxide.

MnO(OH)
2
appears as a brown precipitate. There is some confusion about whether
the oxidised manganese is tetravalent or trivalent. Some sources claim that
Mn(OH)
3
is the brown precipitate, but hydrated MnO
2
may also give the brown
colour.
Iodide ions are added and acidified (acid facilitates the conversion by the brown),
which reduces tetravalent hydroxides back to their stable divalent state thereby
liberating equivalent amount of iodine.

Thiosulphate solution is used, with a starch indicator, to titrate the iodine.

This iodine is equivalent to dissolved oxygen present in the sample.
(2) Electrometric Method
The electrode method offers several advantages over the titrimetric method
including speed, elimination or minimization of interferences, field compatibility,
continuous monitoring and insitu measurement.

Dissolved oxygen can be measured by a special sensor kept in an electrochemical
cell by the amperometric method.

The cell comprises a sensing electrode, a reference electrode and a supporting
electrolyte, a semi-permeable membrane, which served dual function.

It separates the water sample from the electrolyte, and at the same time, permits
only the dissolved oxygen to diffuse from the water sample through the membrane
into the supporting electrolyte.

The diffusion current created by migration of oxygen through a permeable
membrane is linearly proportional to the concentration of molecular oxygen in the
sample.
The diffusion current created by migration of oxygen through a permeable
membrane is linearly proportional to the concentration of molecular oxygen in the
sample.

The sample is treated with manganous sulphate, alkaline-iodide-azide reagent and
finally sulfuric acid. The first two chemicals combine with dissolved oxygen to form
a compound which, when acid is added, releases free iodine (from the potassium
iodide).
1. Burette
2. Burette stand
3. 300 mL glass stoppered BOD bottles
6. Pipette bulb
7. 250 mL graduated cylinders
8. Wash bottle

1. Manganous sulphate solution
2. Alkaline iodide-azide solution
3. Sulfuric acid, Concentrated
4. Starch indicator solution
5. Sodium thiosulphate
6. Distilled or deionized water
7. Potassium Hydroxide
8. Potassium Iodide
9. Sodium Azide

If analysis is to be carried out with in two hours of collection, cool storage is not
necessary. If analysis can not be started with in the two hours of sample collection
to reduce the change in sample, keep all sample at 4 C.
Do not allow samples to freeze. Analysis should begin as shown as possible. Do
not open sample bottle before analysis.
Begin analysis within six hours of sample collection.
10.5.1 PRECAUTIONS
The experiment involves lot of solutions and additions of strong acid and alkali and
hence care should be taken.
Dissolved oxygen concentrations may change drastically depending upon
depth, distance, temperature and period of sampling.
If the sample was obtained by a sampling device of some kind, the water
cannot be simply poured into a BOD bottle, since this would cause aeration
of the sample. Instead, the sample must be drawn off from a tube located
near the bottom of the sampling device. Place the rubber tube into the
bottom of the BOD bottle and fill the bottle, again allowing the bottle to
overflow.
For shallow depth use normal water samplers. However for depth greater
than 150 cm (5 ft), use Kemmerer Sample Bottles.
In the case of electrode method:
Membrane-covered electrode systems minimize the interferences often
encountered with dropping mercury or rotating platinum electrodes.
The sensing element is protected by an oxygen permeable membrane,
which serves as a diffusion barrier against matrix interference problems.
10.6 PROCEDURE:
a) Manganous Sulphate Solution
Dissolve Manganese Sulphate
480g of (or)
400g of (or)
364 g of
in freshly boiled and cooled distilled water, filter the solution and make up to
1000 mL (One litre). In this experiment, we are using Manganese sulphate
Mono hydrate,

Take 364 g Manganese sulphate Mono hydrate ( ) and transfer it
to the beaker. To dissolve the content, place it in the magnetic stirrer.

The solution should not give blue color by addition of acidified potassium
iodide solution and starch.

b) Al kaline Iodide Sodium Azide Solution
To prepare this reagent we are going to mix three different chemicals
Dissolve either
500 g of Sodium Hydroxide (or)
700 g of Potassium Hydroxide and
135 g of Sodium Iodide (or)
150 g of Potassium Iodide
To prepare this reagent, take 700 g of Potassium hydroxide and add 150 g
of potassium iodide and dissolve it in freshly boiled and cooled water, and
make up to 1000 mL (One litre).

Dissolve 10 g of Sodium Azide in 40 mL of distilled water and add
this with constant stirring to the cool alkaline iodide solution prepared.

c) Sodium Thiosulphate Stock Solution

Weigh approximately 25 g of sodium thiosulphate and
dissolve it in boiled distilled water and make up to 1000 mL. Add 1 g of
Sodium Hydroxide to preserve it.

d) Starch Indicator
Weigh 2 g of starch and dissolve in 100 mL of hot distilled water. In case if
you are going to preserve the starch indicator add 0.2 g of salicylic acid as
preservative.

e) Sulphuric Acid

Take two 300-mL glass stoppered BOD bottle and fill it with sample to be
tested. Avoid any kind of bubbling and trapping of air bubbles. Remember
no bubbles!
(Or)
Take the sample collected from the field. It should be collected in BOD bottle
filled upto the rim.
Add 2mL of manganese sulfate to the BOD bottle by inserting the calibrated
pipette just below the surface of the liquid.
Add 2 mL of alkali-iodide-azide reagent in the same manner.
Squeeze the pipette slowly so no bubbles are introduced via the pipette (The
pipette should be dipped inside the sample while adding the above two
reagents. If the reagent is added above the sample surface, you will introduce
oxygen into the sample).
If oxygen is present, a brownish-orange cloud of precipitate or floc will
appear.
Allow it to settle for sufficient time in order to react completely with oxygen.
Add 2 mL of concentrated sulfuric acid via a pipette held just above the
surface of the sample.
Carefully stopper and invert several times to dissolve the floc.
At this point, the sample is "fixed" and can be stored for up to 8 hours if kept
in a cool, dark place.
Rinse the burette with sodium thiosulphate and then fill it with sodium
thiosulphate. Fix the burette to the stand.
Measure out 203 mL of the solution from the bottle and transfer to an conical
flask.
Titration needs to be started immediately after the transfer of the contents to
conical flask.
Titrate it against sodium thiosulphate using starch as indicator. (Add 3 - 4
drops of starch indicator solution)
End point of the titration is first disappearance of the blue color to colorless.
Note down the volume of sodium thiosulphate solution added which gives the
dissolved oxygen in ______ mL
10.7 CALCULATION
For determining the Dissolved Oxygen (DO) in the given water sample, the
readings are required to be tabulated.
10.7.1 TABLE
Burette Solution: Sodium Thiosulphate
Indicator: Starch
End point : Disappearance of blue color
For the calculation of DO the temperature at the time of measurement is 20
C and the volume of sample taken is 200 mL.
sodium thiosulphate is taken in the burette
For the first titration the Initial reading is _____ mL and the final reading is
______ The volume of sodium thiosulphate consumed to get the end point is
______mL.
For the second titration the Initial reading is ______ mL and the final reading
is ______ The volume of sodium thiosulphate consumed to get the end point
is________ mL.
For the third titration the Initial reading is ______mL and the final reading
is ______ The volume of sodium thiosulphate consumed to get the end point
is ______mL.
For the second and third titration, we have achieved concordant value. So we
can go for the calculations.
Trial
No.
Temperature
(C)
Volume of
Sample
(mL)
Burette Reading
(mL) Volume of
Titrant (mL)
Dissolved
Oxygen
(mg/L)
Initial Final
1.
2.
3.
10.7.2 DATA SHEET
DETERMINATION OF DISSOLVED OXYGEN
DATA SHEET
Date Tested :
Tested By :
Project Name :
Sample Number :
Sample Location :
Model Calculation:
Volume of Sodium thiosulphate V
1
=
Normality of Sodium thiosulphate N
1
=
Volume of Sample V
2
=
=
Trial
No.
Temperature
(C)
Volume of
Sample (mL)
Titrant (mL)
Dissolved
Oxygen (mg/L)
Initial Final
1. 20.0 200
2. 20.0 200
3. 20.0 200
The Dissolved Oxygen in the given sample of water at 27C =______mg/L.
10.9 INFERENCE
Dissolved oxygen of the tested sample is ______ mg/L. Test results shows the
water is in healthy condition and fit for aquatic life. IS code does not mentioned
minimum standards for DO. However, for healthy water body, the dissolved oxy-
gen is about _______ parts per million.
10.10 EVALUATION
1. Winkler titration method is based on _____ property of Dissolved Oxygen.
a) Reduction
b) Oxidation
c) Redox
d) Decomposition
2. Dissolved oxygen in the water mainly depends upon Organic content of the
water.

a) True
b) False
3. The ingredients of Alkali are NaOH, NaI
a) NaN
4

b) NaN
3

c) NaN
2

d) NaN

4. The precipitate formed after the addition of MnSO4 and Alkali azide is _______.
a) Manganese Hydroxide
b) Sodium sulphate
c) Potassium sulphate
d) Manganese oxide
5. Dissolved Oxygen depends only on Physical Properties of the water.
a) True
b) False

6. Along the stream the increase in dissolved oxygen in water will be at the

a) riffles
b) warm pool
c) bank erosion
d) top

7. The dissolved Oxygen in potable water_______.

a) imparts freshness
b) improves taste
c) improves smell
d) imparts colour

8. Sulphide and Sulphur dioxide interfere in the determination of dissolved oxygen.

a) True
b) False

9. The sample obtained for testing Dissolved Oxygen can be preserved by
a) adding the reagents and stored at 10 to 20 for up to 8 hours
b) storing at room temperature for up to 24 hours
c) storing at 0 for up to 24 hours
d) adding the reagents and stored at room temperature for up to 24 hours

10. Minimum DO in the fresh water for the survival of aquatic life is
a) 0 mg/l
b) 2 mg/l
c) 8 mg/l
d) 4 mg/l

KEY TO ITEMS:

1) b
2) True
3) b
4) a
5) False
6) a
7) a
8) a
9) a
10) d

11.0 EXPERIMENT ON DETERMINATION OF
BIOCHEMICAL OXYGEN DEMAND
PREAMBLE:
How to determine biochemical oxygen demand in Water and Wastewater.
stated in
th
Edition.
Method 5210 B.
(2) Methods for Chemical Analysis of Water and Wastes, EPA-600/4-79-020, USEPA, Method
405.1.
11.1 AIM
To determine biochemical oxygen demand in the given water sample with the
11.2 INTRODUCTION
The biochemical oxygen demand determination is a chemical procedure for determining
the amount of dissolved oxygen needed by aerobic organisms in a water body to break
the organic materials present in the given water sample at certain temperature over a
specific period of time.
BOD of water or polluted water is the amount of oxygen required for the biological
decomposition of dissolved organic matter to occur under standard condition at a
standardized time and temperature. Usually, the time is taken as 5 days and the
temperature is 20C.
The test measures the molecular oxygen utilized during a specified incubation period for
the biochemical degradation of organic material (carbonaceous demand) and the
oxygen used to oxidize inorganic material such as sulfides and ferrous ion. It also may
measure the amount of oxygen used to oxidize reduced forms of nitrogen (nitrogenous
demand).
BOD is the principle 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.
Efficiency of any treatment plant can be judged by considering influent BOD and the
effluent BOD and so also the organic loading on the unit.
Application of the test to organic waste discharges allows calculation of the effect of the
discharges on the oxygen resources of the receiving water. Data from BOD tests are
used for the development of engineering criteria for the design of wastewater treatment
plants.
Ordinary domestic sewage may have a BOD of 200 mg/L. Any effluent to be discharged
into natural bodies of water should have BOD less than 30 mg/L.
This is important parameter to assess the pollution of surface waters and ground waters
where contamination occurred due to disposal of domestic and industrial effluents.
Drinking water usually has a BOD of 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 stream waters.
The determination of the BOD of wastes is useful in the design of treatment facilities.
It is the only parameter, to give an idea of the biodegradability of any sample and self
purification capacity of rivers and streams.
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.
11.3 PRINCIPLE
The sample is filled in an airtight bottle and incubated at specific temperature for 5 days.
The dissolved oxygen (DO) content of the sample is determined before and after five
days of incubation at 20C and the BOD is calculated from the difference between initial
and final DO.
The initial DO is determined shortly after the dilution is made; all oxygen uptake
occurring after this measurement is included in the BOD measurement.
1. BOD Incubator
2. Burette & Burette stand
3. 300 mL glass stopper BOD bottles
6. Pipette bulb
7. 250 mL graduated cylinders
8. Wash bottle
1. Calcium Chloride
2. Magnesium Sulphate
3. Ferric Chloride
4. Di Potassium Hydrogen Phosphate
5. Potassium Di Hydrogen Phosphate
6. Di sodium hydrogen phosphate
7. Ammonium Chloride
8. Manganous sulphate
9. Potassium hydroxide
10. Potassium iodide
11. Sodium azide
12. Concentrated sulfuric acid
13. Starch indicator
14. Sodium thiosulphate
15. Distilled or deionized

If Analysis is to be carried out within two hours of collection, cool storage is not
necessary. If analysis can not be started with in the two hours of sample collection
to reduce the change in sample, keep all samples at 4

C.
Do not allow samples to freeze. Do not open sample bottle before analysis.
Begin analysis within six hours of sample collection
11.5.1 PRECAUTIONS
Prepare dilution water 3 to 5 days before initiating BOD test to ensure that the
BOD of the dilution water is less than 0.2 mg/L. Discard dilution water if there
is any sign of biological growth
The sample should be adjusted to a pH between 6.5 and 7.5, using sulfuric acid
for samples with pH in the alkaline side i.e., greater than 7.5 or sodium hydroxide
for samples with pH in the acidic side i.e., less than 6.5.
.
Add sodium sulfite (Na
2
SO
3
) to remove residual chlorine, if necessary. Samples
containing toxic metals, arsenic, or cyanide often require special study and
pretreatment.
While still letting sample water flow down the tube, slowly pull the tube from the
bottom of the bottle and fill the bottle to its brim. Check for bubbles. Carefully
stopper the BOD bottle as described above.
11.6 PROCEDURE
11.6.1 PREPARATION OF REAGENT
a) Manganous Sulphate Solution
Dissolve Manganese Sulphate
480g of (or)
400g of (or)
364 g of
in freshly boiled and cooled distilled water, filter the solution and make up to 1000
mL (One litre). In this experiment, we are using Manganese sulphate Mono
hydrate.
Take 364g of and transfer it to the beaker. To dissolve the content,
place it in the magnetic stirrer
Note: The solution should not give blue color by addition of acidified potassium
iodide solution and starch.

b) Al kaline Iodide Sodium Azide Solution
To prepare this reagent we are going to mix three different chemicals
Dissolve either
500 g of Sodium Hydroxide (or)
700 g of Potassium Hydroxide
135 g of Sodium Iodide (or)
150 g of Potassium Iodide

To prepare this reagent, take 700 g of potassium hydroxide and add 150 g of
potassium iodide and dissolve it in freshly boiled and cooled water, and make up
to 1000 mL (One litre).
Dissolve 10 g of Sodium Azide in 40 mL of distilled water and add this
with constant stirring to the cool alkaline iodide solution prepared.
c) Sodium Thiosulphate stock solution
Weigh approximately 25 g of sodium thiosulphate and dissolve
it in boiled distilled water and make up to 1000 mL. Add 1 g of sodium hydroxide
to preserve it.
d) Starch Indicator
Weigh approximately 2 g of starch and dissolve in 100 mL of hot distilled water
e) Sulphuric Acid
.
In case if you are going to preserve the starch indicator add 0.2 g of salicyclic
acid as preservative.

f) Calcium Chloride solution
Weigh accurately 27.5 g of anhydrous calcium chloride and dissolve it in distilled
water.
Transfer it to the 100 mL standard flask and make up to 100 mL using distilled
water.
g) Magnesium Sulphate solution
Weigh accurately 22.5 g of magnesium sulphate and dissolve it in distilled water.
water.
h) Ferric Chloride solution
Weigh accurately 0.15 g ferric chloride and dissolve it in distilled water.
water.
i) Phosphate buffer solution
Weigh accurately 8.5g of Potassium Di Hydrogen Phosphate (KH
2
PO
4
) and
dissolve it in distilled water.
Then add exactly 21.75 g of Di Potassium Hydrogen Phosphate (K
2
HPO
4
) and
dissolve it.
To the same beaker 33.4 g of Di sodium hydrogen phosphate (Na
2
HPO
4
7H
2
O),
is weighed and added.
Finally to the beaker containing all the salts, add accurately 1.7 g of Ammonium
Chloride (NH
4
Cl) and dissolve it.
water.
The pH should be 7.2 without further adjustment.
j) Dilution Water
High quality organic free water must be used for dilution purposes.
The required volume of water (five litres of organic free distilled water) is aerated
with a supply of clean compressed air for at least 12 hours. Allow it to stabilize by
incubating it at 20C for at least 4 hours.
For the test we have taken five litres of organic free aerated distilled water, hence
add 5mL each of the nutrients.
Add 5mL calcium chloride solution
Add 5mL magnesium sulphate solution
Add 5mL ferric chloride solution and
Add 5mL phosphate buffer solution
This is the standard dilution water. Prepare dilution water 3 to 5 days before
initiating BOD test to ensure that the BOD of the dilution water is less than 0.2
mg/L.
Take four 300 mL glass stoppered BOD bottles (two for the sample and two for
the blank).
Add 10 mL of the sample to each of the two BOD bottles and the fill the
remaining quantity with the dilution water. i.e., we have diluted the sample 30
times.
The remaining two BOD bottles are for blank, to these bottles add dilution water
alone.
After the addition immediately place the glass stopper over the BOD bottles and
note down the numbers of the bottle for identification.
Now preserve one blank solution bottle and one sample solution bottle in a BOD
incubator at 20C for five days.
The other two bottles (one blank and one sample) needs to be analysed
immediately.
Avoid any kind of bubbling and trapping of air bubbles. Remember no bubbles!
(The pipette should be dipped inside the sample while adding the above two
reagents. If the reagent is added above the sample surface, you will introduce
oxygen into the sample.)
When this floc has settled to the bottom, shake the contents thoroughly by
turning it upside down.
Add 2 mL of concentrated sulfuric acid via a pipette held just above the surface
of the sample.
Erlenmeyer flask.
Measure out 203 mL of the solution from the bottle and transfer to an Erlenmeyer
flask.
Titrate the solution with standard sodium thiosulphate solution until the yellow
color of liberated Iodine is almost faded out. (Pale yellow color)
Add 1 mL of starch solution.
and continue the titration until the blue color disappears to colourless.
Note down the volume of sodium thiosulphate solution added , which gives the
D.O. in mg/L. Repeat the titration for concordant values.
After five days, take out the bottles from the BOD incubator and analyse the
sample and the blank for DO.
If oxygen is present, a brownish-orange cloud of precipitate or floc will appear.
When this floc has settled to the bottom, shake the contents thoroughly by
turning it upside down.
Add 2 mL of concentrated sulfuric acid via a pipette held just above the surface
of the sample.
Erlenmeyer flask.

Measure out 203 mL of the solution from the bottle and transfer to an Erlenmeyer
flask.
Titrate the solution with standard sodium thiosulphate solution until the yellow
color of liberated Iodine is almost faded out. (Pale yellow color)
Add 1 mL of starch solution and continue the titration until the blue color
disappears to colourless.
Note down the volume of sodium thiosulphate solution added, which gives the
D.O. in mg/L. Repeat the titration for concordant values.
11.7 CALCULATION
For determining the Biochemical Oxygen Demand in the given water sample, the
readings should be tabulated.
11.7.1 TABLE
Burette Solution: Sodium Thiosulphate
Indicator: Starch
End point : Disappearance of blue color
For the calculation of initial DO, immediately after dilution the volume sample
taken is 200 mL.
For the blank titration the burette reading is ______ mL. The volume of titrant
sodium thiosulphate is _______mL. The value of DO in mg/L is ____
For the first titration the burette reading is _______mL. The volume of titrant is
7.9 and the value of DO is ______mg/L.
For the second titration the burette reading is _______ The volume of titrant
is ______mL and the value of DO is _____mg/L.
Trial
No.
Day
Volume of
Sample
(mL)
Burette Reading
(mL)
Volume of
Titrant
(mL)
(Na2S2O3
solution used)
Dissolved
Oxygen
(mg/L) Initial Final
Blank
1.
2.
Blank
1.
2.
For the calculation of DO at the end of five days the volume of sample taken is
200 mL. For the blank titration the value of burette reading is ______. The
volume of titrant is ______mL and the DO is ______ mg/L.
For the first titration the burette reading is ______. The volume of titrant is
______ and the DO value is_______mg/L.
For the second titration the burette reading is _____ The volume of titrant is
_____ and the value of DO is _______ mg/L. We have achieved concordant
values. So we can go for the calculations.
11.7.2 DATA SHEET
DETERMINATION OF BIOCHEMICAL OXYGEN DEMAND
DATA SHEET
Date Tested :
Tested By :
Project Name :
Sample Number :
Sample Location :
Initial DO of the diluted sample, D0 =
DO at the end of 5 days for the diluted sample, D5 =
Blank correction = C0 - C5, BC =
Initial DO of the blank, C0 =
DO at the end of 5 days for the blank, C5 =
Biochemical Oxygen Demand = {D0 D5 BC} x Volume of the diluted sample
Trial
No.
Biochemical Oxygen Demand (mg/L)
Day
Volume of
Sample (mL)
Titrant (mL)
Dissolved
Oxygen (mg/L)
Initial Final
Blank 0 200
3. 0 200
4. 0 200
Blank 5 200
1. 5 200
2. 5 200
The BOD of the given sample of water =______ mg/L.
11.9 INFERENCE
On the basis of the BOD values, the characteristics of the water and the biological
activity of the incubated microflora can be determined. Effluent with high BOD levels is
discharged into a stream or river; it will accelerate bacterial growth in the river and
consume the oxygen levels in the river. The oxygen may diminish to levels that are
lethal for most fish and many aquatic insects. As the river re-aerates due to atmospheric
mixing and as algal photosynthesis adds oxygen to the water, the oxygen levels will
slowly increase downstream. The biological capacity of a sewage treatment plant can
be tested by comparing the BOD value of a known control solution with the BOD
derived from the treatment plant.
BOD detects only the destructible proportion of organic substances and as a general
principle is therefore lower than the COD value, which also includes inorganic materials
and those materials which cannot be biologically, oxidized.
11.10 EVALUATION
1. Biochemical oxygen demand (BOD) is an important measure of
a) the oxygen using potential of water and wastewater
b) oxygen content of water and wastewater
c) an organism's natural level of oxygen requirement
d) a measure of the biological activity of water and wastewater
2. In BOD test, dilution water is aerated
a) for supplementing air
b) for cooling the sample
c) for super saturation
d) for diluting the sample
3. Which of the following is added as nutrient
a) Calcium chloride
b) Calcium sulphate
c) Magnesium chloride
d) Magnesium phosphate
4. Seeding is the process of addition of

a) seeds
b) live microbes
c) cold water
d) nutrients

5. After the incubation period of BOD which is 5 days at 20C,

a) all the organic content would be exhausted.
b) all organisms present will die
c) practical convenience
d) all the nutrients would be exhausted.

6. In a treatment plant when the influent BOD is 245 mg/L and the effluent BOD is 22
mg/L, the percentage of BOD removed is

a) 19%
b) 91%
c) 9%
d) 86%

7. The reaction that occurs between iodine and sodium thiosulphate result in ______.

a) Sodium iodide
b) Disodium iodide
c) Disodium thioiodide
d) Sodium thio iodide

8. Manganous hydroxide takes up dissolved oxygen in molecular form to form
Manganous oxide.

a) Manganous oxide
b) Manganous di oxide
c) Manganic di oxide
d) Manganic oxide

9. Sulphuric acid is added to

a) reduce tetravalent manganese to trivalent manganese
b) reduce tetravalent manganese to divalent manganese
c) reduce tetravalent manganese to manganese
d) make acidic pH

10. The increased level of BOD in water indicate that
a) it is not fit for potable use
b) it is fit for potable use
c) it tastes better
d) it smells pleasant

KEY TO ITEMS:

1) a
2) c
3) a
4) b
5) a
6) b
7) a
8) a
9) b
10) a

12.0 EXPERIMENT ON DETERMINATION OF
CHEMICAL OXYGEN DEMAND
PREAMBLE:
How to determine chemical oxygen demand in Water and Wastewater.
procedure stated in
th
Edition. Method 5220 C.
12.1 AIM
To determine chemical oxygen demand in the given water sample with the
12.2 INTRODUCTION
Before performing this experiment, few questions may arise to the learners:
What is meant by chemical oxygen demand?
Why do we need to determine COD?
What are the methods available to measure COD?
Is it measured in water or wastewater?
Whether is it mandatory to determine COD as per our codal provision?
The chemical oxygen demand (COD) test is commonly used to indirectly
measure the amount of organic compounds in water. Most applications of COD
determine the amount of organic pollutants found in surface water (e.g. lakes and
rivers), making COD a useful measure of water quality. It is expressed in
milligrams per liter (mg/L), which indicates the mass of oxygen consumed per
liter of solution.
COD is the measurement of the amount of oxygen in water consumed for
chemical oxidation of pollutants.
COD determines the quantity of oxygen required to oxidize the organic matter in
water or waste water sample, under specific conditions of oxidizing agent,
temperature, and time.
This method covers the determination of COD in ground and surface waters,
domestic and industrial wastewaters. The applicable range is 3-900 mg/L.

COD values are particularly important in the surveys designed to determine and
control the losses to sewer systems.

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
that wastewater highly polluted and amenable to the biological treatment.

It is useful to assess strength of wastes, which contain toxins and biologically
resistant organic substances.

COD can be related to TOC, however, does not account for oxidation state of the
organic matter.

BOD value is always lower than COD value. For domestic and some industrial
wastewater, COD value is about 2.5 times BOD value.
12.3 PRINCIPLE
The organic matter present in sample gets oxidized completely by potassium
dichromate (K
2
Cr
2
O
7
) in the presence of sulphuric acid (H
2
SO
4
), silver sulphate
(AgSO
4
) and mercury sulphate (HgSO
4
) to produce CO
2
and H
2
O. The sample is
refluxed with a known amount of potassium dichromate (K
2
Cr
2
O
7
) in the sulphuric
acid medium and the excess potassium dichromate (K
2
Cr
2
O
7
) is determined by
titration against ferrous ammonium sulphate, using ferroin as an indicator. The
dichromate consumed by the sample is equivalent to the amount of O
2
required
to oxidize the organic matter.

1. COD Digester
2. Burette & Burette stand
3. COD Vials with stand
4. 250 mL conical flask (Erlenmeyer Flask)
5. Pipettes
6. Pipette bulb
7. Tissue papers
8. Wash Bottle

1. Potassium dichromate
2. Sulfuric acid
3. Ferrous ammonium sulphate
4. Silver sulphate
5. Mercury sulphate
6. Ferroin indicator
7. Organic free distilled water

Samples are collected in glass bottles. Use of plastic containers is permitted if it
is known that there is no organic contaminants present in it.

Biologically active samples should be tested as soon as possible. Samples
containing settleable material should be well mixed, preferably homogenized, to
permit removal of representative aliquots.

Samples should be preserved with sulphuric acid to a pH <2 and maintained at
4
0
C until analysis.

Do not allow the samples to freeze.

12.5.1 PRECAUTIONS
Chlorides are quantitatively oxidized by dichromate and represent a
positive interference. Mercuric sulfate is added to the digestion tubes to
complex the chlorides so that it does not interfere in the determination.

Nitrites also interfere in the determination of COD and hence during the
determination of samples with high concentration of nitrites, 120mg of
sulphuric acid is added to the potassium dichromate solution.

Traces of organic material either from the glassware or atmosphere may
cause a positive error. Extreme care should be exercised to avoid
inclusion of organic materials in the distilled water used for reagent
preparation or sample dilution.

12.6 PROCEDURE
a) Standard Potassium Dichromate Reagent - Digestion Solution
Weigh accurately 4.913 g of potassium dichromate, previously dried at
103C for 2 - 4 hours and transfer it to a beaker.

Weigh exactly 33.3g of mercuric sulphate and add to the same beaker.
Measure accurately 167 mL of concentrated sulphuric acid using clean dry
measuring cylinder and transfer it to the beaker. Dissolve the contents and
cool to room temperature. (If not dissolved keep it over night).


Carefully transfer the contents to the 1000 mL standard flask and make up
to 1000 mL using distilled water.

This is the standard potassium dichromate solution to be used for
digestion.
b) Sulphuric Acid Reagent - Catal yst Solution
Weigh accurately 5.5 g silver sulphate crystals to a dry clean 1000 mL
beaker. To this carefully add about 500 mL of concentrated sulphuric acid
and allow to stand for 24 hours (so that the silver sulphate crystals
dissolve completely).

c) Standard Ferrous Ammonium Sulphate solution
Weigh accurately 39.2g of ferrous ammonium sulphate crystals and
dissolve it in distilled water.


Carefully transfer the contents to the 1000 mL standard flask and make up
to 1000 mL mark using distilled water.

Take three COD vials with stopper (two for the sample and one for the
blank).
Add 2.5 mL of the sample to each of the two COD vials and the remaining
COD vial is for blank; to this COD vial add distilled water.
Add 1.5 mL of potassium dichromate reagent - digestion solution to each
of the three COD vials.
Add 3.5 mL of sulphuric acid reagent - catalyst solution in the same
manner.
CAUTION: COD vials are hot now.
Cap tubes tightly. Switch on the COD Digester and fix the temperature at
150 C and set the time at 2 hours.
Place the COD vials into a block digester at 150C and heat for two hours.
The digester automatically switches off. Then remove the vials and allow
it to cool to the room temperature.
Meanwhile, get ready with the burette for the titration.

Fill the burette with the ferrous ammonium sulphate solution, adjust to
zero and fix the burette to the stand.
Transfer the contents of the blank vial to conical flask.
Add few drops of ferroin indicator. The solution becomes bluish green in
colour.
Titrate it with the ferrous ammonium sulphate taken in the burette.
End point of the titration is the appearance of the reddish brown colour.
Note down the volume of ferrous ammonium sulphate solution added for
the blank (A) is _______ mL.
Transfer the contents of the sample vial to conical flask.
Add few drops of ferroin indicator. The solution becomes green in colour.
Titrate it with the ferrous ammonium sulphate taken in the burette.
End point of the titration is the appearance of the reddish brown colour.
Note down the volume of ferrous ammonium sulphate solution added for
the sample (B) is _________mL.
12.7 CALCULATION
For determining the Chemical Oxygen Demand in the given water sample, the
readings should be tabulated.
12.7.1 TABLE
Burette Solution: Ferrous Ammonium Sulphate
Indicator: Ferroin Indicator
End point: Appearance of reddish brown color
For the blank titration the volume of sample taken is _______ mL.
Ferrous Ammonium Sulphate is taken in the burette.
The obtain reading is ________mL. Similarly for sample one the vol-
ume of sample taken is _______ mL.
Ferrous Ammonium Sulphate is taken in the burette
Sl No. Sample
Volume of
Sample
(mL)
Burette Reading
(mL)
Volume of 0.1 N
FAS (mL)
Initial Final
1.

2.
3.
The initial reading is _______mL and the final reading is ______ mL.
Volume of Ferrous Ammonium Sulphate consumed to get the end point is
______ mL.
For sample two the initial reading is _______mL and the final reading is
________mL.
The volume of Ferrous Ammonium Sulphate consumed to get the end
point is _______mL.
For sample 1 and 2 the reading as same so we can go for the calculations.
12.7.2 DATA SHEET
DETERMINATION OF CHEMICAL OXYGEN DEMAND
DATA SHEET
Date Tested :
Tested By :
Project Name :
Sample Number :
Sample Location :
Volume of Ferrous Ammonium sulphate for blank (A) =
Volume of Ferrous Ammonium sulphate for Sample (B) =
Normality of Ferrous Ammonium sulphate N =
Volume of Sample V =
Chemical Oxygen Demand =
Sl No.
(A - B * N * 8 * 1000)
To convert the sample size from mL to L, multiply the result by 1,000 mL/L to convert the sample size
from mL to L.
Residual Chlorine (mg/L) =
=
Sample
Volume of
Sample (mL)
Burette Reading (mL) Volume of 0.1 N
FAS (mL)
Initial Final
4. Blank 2.5
5. Sample 1 2.5
6. Sample 2 2.5
The COD of the given sample of water =_______mg/L.
12.9 INFERENCE
Chemical oxygen demand does not differentiate between biologically available
and inert organic matter, and it is a measure of the total quantity of oxygen
required to oxidize all organic material into carbon dioxide and water. COD
values are always greater than BOD values. For domestic and some industrial
wastewater COD is about 2.5 times BOD.
12.10 EVALUATION
1. Potassium dichromate is considered as the best
a) Oxidizing agent
b) Reducing agent
c) Redox agent
d) Chemical agent

2. Mercury Sulphate is added to reduce the interference of
a) Chlorides.
b) Sulphates
c) Organic pollutants
d) Hardness
3. Silver Sulphate is added as
a) Oxidizing agent
b) Reducing agent
c) Redox agent
d) Catalyst
4. Ferroin indicator is
a) Phenanthroline mono hydrate
b) Ferric sulphate
c) Phenanthroline mono hydrate and Ferric Sulphate
d) Ferrous Sulphate
5. After refluxing, ___________ solution is titrated against FAS.

a) excess potassium dichromate
b) consumed potassium dichromate
c) initially added potassium dichromate
d) potassium dichromate and silver sulphate

6. H
2
SO
4
is added to FAS solution
a) as it is a component of the reagent
b) to prevent hydrolysis of ferrous sulphate into ferrous hydroxide
c) to provide acidic medium
d) to neutralise the medium

7. The products formed after COD analysis are ______.

a) Carbon di oxide and water
b) Water alone
c) Carbon di oxide alone
d) Carbon monoxide and water

8. In industrial waste water, COD value is about _____________ BOD value.

a) 2.5 times
b) 3.5 times
c) 4.5 times
d) 5.5 times

9. Sulphuric acid is added

a) as it assists in oxidizing the nitrogen compounds
b) to provide acidic medium
c) to neutralise the medium
d) as catalyst

10. A blank solution is
a) identical in all respects to the test solution except for the absence of
test solute
b) identical in all respects to the test solution
c) a solution without any reagents
d) a solution without distilled water

KEY TO ITEMS:

1) a
2) a
3) d
4) c
5) a
6) b
7) a
8) a
9) a
10) a

13.1 DETERMINATION OF IRON
Aim
To determine the quantity of iron present in the given sample of water.
Principle
Iron is usually present in natural water and is not objectionable, if concentration is less than 0.3 ppm. It may be in
true solution in colloidal state that may be peptized by organic matter, in the inorganic and organic iron complexes,
or in relatively coarse suspended particles. It may be either ferrous or ferric, suspended or filterable. Iron exists in
soils and minerals mainly as insoluble ferric oxide and iron sulphide (pyrite). It occurs in some areas, also as ferrous
carbonate (siderite), which is very slightly soluble.
The phenanthroline method is the preferred standard procedure for the measurement of iron in water except
when phosphate or heavy metal interferences are present. The method depends upon the fact that 1,
10-phenanthroline combine with Fe
++
to form an orange-red complex. Its colour conforms to Beers law and is
readily measured by visual or photometric comparison. Small concentration of iron can be most satisfactorily
determined by colorimetric analysis. It is also based on Beers law. By measuring the intensities of transmitted and
incident light through a coloured solution and knowing its optical density or transmission, we can prepare a calibration
curve and subsequent concentration can be read.
Phenanthroline Method
Apparatus
1. Colorimetric 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.
2. Glassware like conical flasks, pipettes and glass beads.
13.0 DETERMINATION OF IRON
Reagents
1. Hydrochloric acid 2. Hydroxylamine solution
3. Ammonium acetate buffer solution 4. Sodium acetate solution
5. Phenanthroline solution 6. Stock iron solution
7. Standard iron solution (1 mL = 10 g Fe)
Procedure
1. Pipette 10, 20, 30 and 50 mL. Standard iron solution into 100 mL conical flasks.
2. Add 1 mL hydroxylamine solution and 1 mL sodium acetate solution to each flask.
3. Dilute each to about 75 mL with distilled water.
4. Add 10 mL phenanthroline solution to each flask.
5. Make up the contents of each flask exactly to 100mL by adding distilled water and left stand for 10
minutes.
6. Take 50 mL distilled water in another conical flask.
7. Repeat steps 2 to 5 described above.
8. Measure the absorbance of each solution in a spectrophotometer at 508 nm against the reference blank
prepared by treating distilled water as described in steps 6 and 7. Prepare a calibration graph taking
meter reading on y-axis and concentration of iron on x-axis.
9. For visual comparison, pour the solution in 100 mL tall form Nessler tubes and keep them in a stand.
10. Mix the sample thoroughly and measure 50 mL into a conical flask.
11. Add 2 mL conc. hydrochloric acid (HCl) and 1mL hydroxylamine solution. Add a few glass beads and
heat to boiling. To ensure dissolution of all the iron, continue boiling until the volume is reduced to 15
to 20 mL.
12. Cool the flask to room temperature and transfer the solution to a 100 mL Nessler tube.
13. Add 10 mL ammonium acetate buffer solution and 2 mL phenanthroline solution and dilute to the 100
mL mark with distilled water.
14. Mix thoroughly and allow at least 10 to 15 minutes for maximum colour development.
15. Measure the absorbance of the solution in a 1cm cell in a spectrophotometer at 508 nm.
16. Read off the conc. of iron (mg Fe) from the calibration graph for the corresponding meter reading.
17. For visual comparison, match the colour of the sample with that of the standard prepared in steps 1
to 7 above.
18. The matching colour standard will give the concentration of iron in the sample (g Fe).
Observation
Standard iron solution in mL Iron content in g Absorbance
Sample calculation
iron (Fe) in mg/L = g Fe/mL of sample
= ..... mg/L
Results
Sample no. or description Iron content in mg/L (Fe)
Discussion
Sample no. Absorbance Iron content from graph in g Iron as Fe in mg/L
Aim
To determine the ammonia nitrogen of the given sample of water.
Principle
Colorimetric method, using Nesslers reagent is sensitive to 20mg/L of ammonia N and may be used up to 5mg/L
of ammonia N. Turbidity, colour and substances precipitated by hydroxyl ion interfere with the determination. The
sample containing ammonia must be analysed immediately after collection; if not 0.8 M conc. H
2
SO
4
/L should be
added to the sample stored at 4C.
Direct Nesslerisation
Direct Nesslerisation is used only for purified water, natural water and highly purified effluents, which have low
ammonia concentration. In samples that have been properly clarified by a pretreatment method using zinc sulphate
and sodium hydroxide, it is possible to obtain a measure of the amount of ammonia N by treatment with Nesslers
reagent, which is strongly alkaline solution of potassium mercuric iodide (K
2
HgI
4
). It combines with NH
3
in alkaline
solution to form a yellowish brown colloidal dispersion, whose intensity of colour is directly proportional to the
amount of NH
3
present. The yellow colour or reddish brown colour typical of ammonia N can be measured in a
spectrophotometer in the wavelength of 400500 nm with a light path of 1cm.
Apparatus
1. Spectrophotometer, or Nessler tube tall form (50 mL or 100 mL capacity)
2. pH meter
Reagents
1. Zinc sulphate solution 2. EDTA reagent as stabiliser
3. Nesslers reagent 4. Stock ammonium solution 1.00 mL = 1.00 mg
14.0 AMMONIA NITROGEN
Procedure
1. Residual chlorine is removed by means of a dechlorinating agent (one or two drops sodium thiosulphate
solution)
2. 100 mL ZnSO
4
solution is added to 100 mL sample and to it is added 0.5mL of NaOH solution to
obtain a pH of 10.5. This is mixed thoroughly.
3. The floc formed is allowed to settle and the clear supernatent is taken for Nesslerisation.
4. If the sample contain Ca or Mg, EDTA reagent is added to 50mL of sample.
5. To this is added 2 mL of Nesslers reagent (proportional amount to be added (if the sample volume
is less).
6. A blank using distilled ammonia free water is treated with Nesslers reagent as above. The absorbance
is fixed as zero.
7. Then the sample is put in 1cm standard tubes of spectrophotometer and the absorbance noted at 400
500nm wavelengths.
8. A calibration curve is prepared as follows:
With 0, 0.2, 0.4, 0.7, 1.0, 1.4, 1.7, 2.0, 2.5, 3.0, 4.0, 5.0 mL of standard NH
4
Cl solution in 50 mL
distilled water standard diluted samples are prepared.
9. Each sample is Nesslerised as indicated earlier and the absorbance is noted down.
10. A graph with mg of NH
3
along x-axis and absorbance along y-axis is plotted and a straight-line graph
is drawn.
11. From the absorbance of a solution of unknown concentration, the g of NH
3
present can be read from
the calibration curve.
Calculation
ammonia N in mg/L =
A
mL of sample
where, A = g N found colorimetrically
Observation
The observation is presented in Tables A and B respectively.
Table A: Observation for calibration
Stock ammonia solution in mL Ammonia Absorbance
Table B
Results
Discussion
Sample no. Absorbance Ammonia nitrogen in g Ammonia nitrogen in mg
from graph
Sample no. or description Ammonia nitrogen in mg/L
Questions
1. Discuss the significance of ammonia nitrogen in water.
2. What is the source of ammonia nitrogen in water?
Aim
To determine the nitrate nitrogen of the given sample of water.
Principle
The reaction with the nitrate and brucine produces yellow colour that can be used for the colorimetric estimation of
nitrate. The intensity of colour is measured at 410 nm. The method is recommended only for concentration of 0.1
2.0 mg/L
3
NO
N. All strong oxidising and reducing agent interfere. Sodium arsenite is used to eliminate interference
by residual chlorine; sulphanilic acid eliminates the interferences by
2
NO
N and chloride interference is masked
by addition of excess NaCl. High concentration of organic matter also may interfere in the determination.
Apparatus
1. Spectrophotometer 2. Water bath
3. Reaction tubes 4. Cool water bath
Reagents
1. Stock nitrate solution 2. Standard nitrate solution
3. Sodium arsenite solution 4. Brucine-sulphanilic acid solution
5. Sulphuric acid solution 6. Sodium chloride solution
Procedure
1. Nitrate standards are prepared in the range 0.11.0 mg/LN diluting 1.00, 2.00, 4.00, 7.00 and
10.0 mL standard nitrate solution to 10 mL with distilled water.
2. If residual chlorine is present 1 drop of sodium arsenite solution is added for each 0.1 mg Cl
2
and mixed.
3. Set up a series of reaction tubes in test tube stand. Add 10 mL sample or a portion diluted to 10 mL
to the reaction tubes.
4. Place the stand in a cool water bath and add 2 mL NaCl solution and mix well.
5. Add 10 mL H
2
SO
4
solution and again mix well and allow cooling.
15.0 NITRATE NITROGEN
6. The stand is then placed in a cool water bath and add 0.5 ml brucine-sulphanilic acid reagent. Swirl
the tubes and mix well and place the tubes in boiling water bath at temperature 95C.
7. After 20 minutes, remove the samples and immerse in cool water bath.
8. The sample are then poured into the dry tubes of spectrophotometer and read the standards and sample
against the reagent blank at 410 nm.
9. Prepare a standard curve for absorbance value of standards (minus the blank) against the concentration
of
3
NO N.
10. Read the concentration of
3
NO
N in the sample from the known value of absorbance.
Calculation
Nitrate N in mg/L =
3
g NO N
mL sample
NO
3
in mg/L = mg/L nitrate N 4.43.
Observation
The observation are presented in Tables A and B respectively.
Table B
Stock nitrate solution in mL Nitrate Absorbance
Sample no. Absorbance Nitrate nitrogen in g from graph Nitrate nitrogen in mg
Results
Discussion
Questions
1. In what forms does nitrogen normally occur in natural waters?
2. Discuss the significance of nitrate nitrogen analysis in water pollution control.
3. Differentiate between nitrite nitrogen and nitrate nitrogen.
4. Discuss the application of nitrate nitrogen data.
5. What are the various methods available for the determination of nitrate nitrogen?
Sample no. or description Nitrate nitrogen in mg/L
Aim
To determine the nitrite nitrogen of the given sample of water.
Principle
The nitrite concentration is determined through the formation of a reddish-purple azo dye produced at pH 2.02.5
by the coupling of diazotised sulphanilic acid with N-(1-naphthyl)-ethylenediamine dihydrochloride.
Apparatus
1. Spectrophotometer
Reagents
1. Sulphanilamide reagent
2. N-(1-naphthyl)-ethylenediamine dihydrochloride solution
3. Hydrochloric acid (1+3)
4. Stock nitrite solution
5. Standard nitrite solution
Procedure
1. To 50ml clear sample neutralised to pH 7, add 1ml sulphanilamide solution.
2. Allow the reagent to react for a period of 28 minute.
3. Then add 0.1ml of 1-naphthyl ethylenediamine solutions and mix immediately.
4. Measure the absorbance of the solution after 10 minute at 543nm at 1cm light path.
5. Prepare standard calibration curve as in any other case.
6. By noting the absorbance of an unknown sample, the concentration of nitrate can be determined.
16.0 NITRITE NITROGEN
Calculation
Nitrite N in mg/L =
mg Nitrite N
mL of sample
Observation
Table B
Results
Stock nitrite solution in mL Nitrite Absorbance
Sample no. Absorbance Nitrite nitrogen in g from graph Nitrite nitrogen in mg
Sample no. or description Nitrite nitrogen in mg/L
Discussion
Questions
1. Explain why sensitive colorimetric methods are needed for the determination of nitrite nitrogen.
2. Explain the nitrogen cycle.
3. What is the significance of determination of nitrite nitrogen in water?
Aim
To determine the Kjeldahl nitrogen of the given sample of water.
Principle
In the presence of sulphuric acid, potassium sulphate and mercuric sulphate catalyst, the amino nitrogen of many
organic materials is converted to ammonium sulphate. After the mercury-ammonium complex, the digestible has
been decomposed by sodium thiosulphate, the ammonia is distilled from an alkaline medium and absorbed in boric
acid. The ammonia is determined colorimetrically or by titration with a standard mineral acid.
Apparatus
1. Digestion apparatus of 800mL capacity
2. Distillation apparatus
3. Spectrophotometer
Reagent
1. All reagents listed for the determination of ammonia N
2. Digestion reagent
3. Phenolphthalein indicator
4. Sodium hydroxide-sodium thiosulphate reagent
5. Borate buffer solution
6. Sodium hydroxide 6N
Procedure
1. Place a measured sample into a digestion flask. Dilute the sample to 300 mL, and neutralise to pH7.
Sample size is determined as follows:
17.0 KJELDAHL NITROGEN
Sample size determination
Organic nitrogen in sample (mg/L) Sample size (mL)
0-1 500
1-10 250
10-20 100
20-50 50
50-100 25
2. Add 25 mL borate buffer and 6 N NaOH until pH 9.5 is reached.
3. Add a few glass beads and boil off 300 mL.
4. Cool and add carefully 50 mL digestion reagent. After mixing heat under a hood until the solution clean
to a pale straw colour.
5. Digest for another 30 minute and allow the flask and contents cool.
6. Dilute the contents to 300 mL and add 0.5 mL phenolphthalein solution.
7. Add sufficient hydroxide-thiosulphate reagent to form an alkaline layer at the bottom of the flask.
8. Connect the flashed to the steamed out distillation apparatus and more hydroxide-thiosulphate reagent.
If a red phenolphthalein colour fails to appear at this stage.
9. Distilled and collect 200 mL distillate below the surface of boric acid solution. Extend the lip of condenser
well bellow the level of boric acid solution.
10. Determine the ammonia as described earlier by taking 50 mL portion of the distillate.
11. Carry out a similar procedure for a blank and apply the necessary correction.
Observation
Stock ammonia solution in mL Ammonia Absorbance
Table B
Calculation
Organic N in mg/L =
A1000
mL of sample
B and C
where, A = mg N found colorimetrically
B = mL of total distillate collected including H
3
BO
3
C = mL of distillation taken for Nesslerisation.
Results
Discussion
Questions
1. What is the difference between Kjeldahl nitrogen and albuminoidal nitrogen?
2. In which form the organic nitrogen exists in domestic wastewater?
Sample no. Absorbance Ammonia nitrogen in g from graph Ammonia nitrogen in mg
Sample no. or description Organic nitrogen in mg/L
18.0 JAR TEST FOR DETERMINING
OPTIMUM COAGULANT DOSAGE
Aim
To determine the optimum coagulant dosage for clarifying the given sample of water by using alum as the coagulant
and performing the jar test experiment.
Principle
Coagulants are used in water treatment plants
(i) to remove natural suspended and colloidal matter,
(ii) to remove material which do not settle in plain sedimentation, and
(iii) to assist in filtration.
Alum [Al
2
(SO
4
)
3
. 18H
2
O] is the most widely used coagulant. When alum solution is added to water, the
molecules dissociate to yield
2
4
SO

and Al
3+
. The +ve species combine with negatively charged colloidal to neutralise
part of the charge on the colloidal particle. Thus, agglomeration takes place. Coagulation is a quite complex
phenomenon and the coagulant should be distributed uniformly throughout the solution. A flash mix accomplishes
this.
Jar test is simple device 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 20 to 30
minutes, corresponding to the flash mixing and slow mixing in the flocculator of the treatment plant. After 30
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
1. Jar test apparatus 2. Glass beakers
3. Pipette 4. Nephelometer
5. pH meter
Reagents
1. Alum solution (1mL containing 10mg of alum)
2. Lime
3. Acid/alkali
Procedure
1. Take 1-litre beakers and fill them with sample up to the mark.
2. Keep each beaker below each paddle and lower the paddles, such that each one is about 1cm above
the bottom.
3. Find the pH of the sample and adjust it to 6 to 8.5.
4. Pipette 1, 2, 3, 4, 5, 6 mL of the alum solution into the test samples.
5. Immediately run the paddles at 100 rpm for 1 minute.
6. Reduce the speed to 3040 rpm and run at this rate for 30 minutes.
7. Stop the machine, lift out the paddles and allow to settle for 30 minutes.
8. Find the residual turbidity of the supernatant using nephelometer.
9. Plot a graph with alum dosage along x-axis and turbidity along y-axis.
10. The dosage of alum, which represents least turbidity, gives Optimum Coagulant Dosage (O.C.D.).
11. Repeat steps 110 with higher dose of alum, if necessary.
Observation
Trial no. Alum dosage in mg/L Turbidity in NTU
Results
Optimum coagulant dosage = . .........
Discussion
Questions
1. Why is alum preferred to other coagulants?
2. What is the difference between coagulation and flocculation?
3. What are coagulant aids?
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?
APPENDIXI. PREPARATION OF REAGENTS AND MEDIA
Reagents for various determinations are prepared as follows:
Alkalinity
1. 0.02 N standard sulphuric acid: Prepare stock solution approximately 0.1 N by diluting 2.5 mL
concentrated sulphuric acid to 1 litre. Dilute 200 mL of the 0.1 N stock solution to 1 litre CO
2
free
distilled water. Standardise the 0.02 N acid against a 0.02 N sodium carbonate solution which has been
prepared by dissolving 1.06 g anhydrous Na
2
CO
3
and diluting to the mark of a 1 litre volumetric flask.
2. Methyl orange indicator: Dissolve 500 mg methyl orange powder in distilled water and dilute it to
1 litre. Keep the solution in dark or in an amber coloured bottle.
3. Phenolphthalein indicator: Dissolve 5 g phenolphthalein in 500mL ethyl alcohol and add 500 mL
distilled water. Then add 0.02 N sodium hydroxide drop-wise until a faint-pink colour appears.
4. Sodium thiosulphate 0.1 N: Dissolve 25 g Na
2
S
2
O
3
.5H
2
O and dilute to 1 litre.
Chloride
5. Potassium chromate indicator: Dissolve 50 g potassium chromate (K
2
Cr
2
O
4
) in a little distilled water.
Add silver nitrate solution until a definite red precipitate is formed. Let stand for 12 hours, filter and
dilute the filtrate to 1 litre with distilled water.
6. Standard silver nitrate solution 0.0141 N: Dissolve 2.395 g AgNO
3
in distilled water and dilute
to 1 litre. Standardise against 0.0141 N NaCl. Store in a brown bottle; 1 mL = 500 g Cl
2
.
7. Standard sodium chloride 0.0141N: Dissolve 824.1 mg NaCl (dried at 140C) in chloride free
water and dilute to 1 litre. 1mL = 500 g Cl
2
.
8. Aluminium hydroxide suspension: Dissolve 125 g aluminium potassium sulphate in 1 litre water.
Warmto 60C and add 55 mL concentrated NH
4
OH slowly with stirring. Let stand for 1 hour,
transfer the mixture to a large bottle. When freshly prepared the suspension occupies a volume of
approximately 1 litre.
Iron
9. Hydrochloric acid: Concentrated HCl.
10. Hydroxylamine solution: Dissolve 10 g hydroxylamine hydrochloride salt (NH
2
OH.HCl) in 100
mL distilled water.
11. Ammonium acetate buffer solution: Dissolve 250 g ammonium acetate (NH
4
C
2
H
3
O
2
) in 150
mL distilled water. Add 700 mL concentrated (glacial) acetic acid.
12. Sodium acetate solution: Dissolve 200 g sodium acetate (NaC
2
H
3
O
2
.3H
2
O) in 800 mL distilled water.
13. Phenanthroline solution: Dissolve 100 mg 1, 10-phenanthroline monohydrate (C
12
H
8
N
2
.H
2
O) in
100 mL distilled water by stirring and heating to 80C. Do not boil. Discard the solution if it darkens.
Heating is unnecessary if 2 drops of concentrated HCl are added to the distilled water. 1 mL of
this reagent is sufficient for no more than 100 g Fe.
APPENDICES
14. Stock iron solution: Add slowly 20 mL concentrated H
2
SO
2
to 50 mL distilled water and dissolve
1.404 g ferrous ammonium sulphate [Fe(NH
4
)
2
(SO
4
)
2
.6H
2
O]. Add 0.1 N KMnO
4
drop wise until a
faint-pink colour persists. Dilute to 1litre with iron free distilled water. Each 1 mL of this solution contains
200 g Fe.
15. Standard iron solution: Pipette 50 mL stock solution into 1 litre volumetric flask and dilute to the mark
with distilled water. 1 mL = 10 g Fe.
Dissolved oxygen
16. Manganous sulphate solution: Dissolve 480 g MnSO
4
.4H
2
O, 400 g MnSO
2
.2H
2
O or 364 g
MnSO
4
.H
2
O in distilled water, filter and dilute to 1 litre.
17. 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 litre. Add 10 g sodium azide (NaN
3
) dissolved in 40 mL
distilled water. The reagent should not give colour with starch when diluted and acidified.
18. Sulphuric acid concentrated: 1mL is equivalent to about 3 mL alkali-iodide-azide reagent.
19. Standard sodium thiosulphate 0.025 N: Dissolve 6.205 g sodium thiosulphate (Na
2
S
2
O
3
.5H
2
O) in
freshly boiled and cooled distilled water and dilute to 1 litre. Preserve by adding 5 mL chloroform or
0.4 g NaOH/L or 4 g borax and 510 mg HgI
2
/L. Standardise this with 0.025 N potassium dichromate
solution which is prepared by dissolving 1.226 g potassium dichromate in distilled water and diluted to
1 litre.
20. Standard potassium dichromate solution 0.025 N: A solution of potassium dichromate equivalent
to 0.025 N sodium thiosulphate contains 1.226 g/L K
2
Cr
2
O
7
. Dry K
2
Cr
2
O
7
at 103C for 2 hrs before
making the solution.
21. Standardisation of 0.025 N sodium thiosulphate solution: Dissolve approximately
2 g KI in an Erlenmeyer flask with 100 to 150 mL distilled water. Add 10 mL of H
2
SO
4
, followed
by exactly 20 mL, 0.1 N potassium dichromate solution. Place in the dark for 5 minutes, dilute to
approximately 400 mL and titrate with 0.025 N sodium thiosulphate solution, adding starch towards
the end of titration. Exactly 20 ml 0.025 N thiosulphate will be consumed at the end of the titration.
Otherwise, the thiosulphate solution should be suitably corrected.
22. Starch Indicator: Add cold water suspension of 5 g soluble starch to approximately 800 mL boiling
water with stirring. Dilute to 1 litre, allow to boil for a few minutes and let settle overnight. Use supernatant
liquor. Preserve with 1.25 g salicylic acid/1 litre or by the addition of a few drops of toluene.
BOD
36. Phosphate buffer solution: Dissolve 8.5 g potassium dihydrogen phosphate (KH
2
PO
4
), 21.75 g
dipotassium hydrogen phosphate (K
2
HPO
4
), 33.4 g disodium hydrogen phosphate heptahydrate
(Na
2
HPO
4
.7H
2
O) and 1.7 g NH
4
Cl in about 500 ml distilled water and dilute to 1 litre. The pH of
this buffer should be 7.2 without further adjustment. Discard the reagent if there is any sign of biological
growth in the stock bottle.
37. Magnesium sulphate solution: Dissolve 22.5 g MgSO
4
.7H
2
O in distilled water and dilute to 1 litre.
38. Calcium chloride solution: Dissolve 27.5 g anhydrous CaCl
2
in distilled water and dilute to 1 litre.
39. Ferric chloride solution: Dissolve 0.25 g FeCl
3
.6H
2
O in distilled water and dilute to 1 litre.
40. Sodium sulphate solution 0.025 N: Dissolve 1.575 g anhydrous Na
2
SO
3
in 1 litre distilled water.
This is to be prepared daily.
41. Seeding: The standard seed material is settled domestic wastewater that has been stored at 20C for
24 to 36 hours. A seed concentration of 12 mL/L is usually adopted.
Coliform test
43. Lactose broth: Beef extract 3 g, peptone 5 g, lactose 5 g and reagent grade distilled water 1 litre.
Add these ingredients to reagent grade distilled water, mix thoroughly and heat to dissolve. pH should
be 6.87.0 after sterilisation.
44. Lauryl tryptose broth: Tryptose 20 g, lactose 5 g, K
2
HPO
4
2.75 g, KH
2
PO
4
2.75 g, NaCl 5 g,
sodium lauryl sulphate 0.1 g, reagent grade distilled water 1 litre, sterilise and use. Add dehydrated
ingredients to water, mix thoroughly and heat to dissolve. pH should be 6.8 2 after sterilisation.
45. Endo agar: Peptone 10 g, lactose 10 g, K
2
HPO
4
3.5 g, agar 15 g, sodium sulphite 2.5 g, basic fuchsin
0.5 g, distilled water 1 litre, pH 7.4 after sterilisation.
46. EMB agar: Peptone 10 g, lactose 10 g, K
2
HPO
4
2 g, agar 15 g, eosin 0.4 g, methylene blue
0.065 g, distilled water 1 litre, pH should be 7.1 after sterilisation.
47. Brilliant green lactose bile broth: Peptone 10 g, lactose 10 g, oxgall 20 g, brilliant green 0.0133 g,
distilled water 1 litre, pH should be 7.2 after sterilisation and is then ready for use. Store away from
direct sunlight to extend the reagent stability to 6 months.
Acidity
48. NaOH solution 0.02 N: Dissolve 4 g NaOH in 1 litre water. This gives 0.1 N NaOH solution. Take
200 ml of this 0.1 N solution and make it up to 1 litre to obtain 0.02 N NaOH solution.
Appendices
Appendices
49. Methyl orange indicator: Dissolve 500 mg methyl orange powder in distilled water and dilute it to
1 litre.
50. Phenolphthalein indicator: Dissolve 5 g phenolphthalein disodium salt in distilled water and dilute to
1 litre.
51. Sodium thiosulphate 0.1 N: Dissolve 25 g Na
2
S
2
O
3
.5H
2
O and dilute to 1 litre distilled water.
COD
52. Standard potassium dichromate solution 0.25 N: Dissolve 12.259 g K
2
Cr
2
O
7
primary standard
grade previously dried at 103C for 2 hours and dilute to 1 litre.
53. Sulphuric acid reagent: Concentrated H
2
SO
4
containing 22 g silver sulphate per 4 kg bottle. Dissolve
22 g Ag
2
SO
2
in 4 kg bottle and keep it for 2 days. This is the reagent.
54. Standard ferrous ammonium sulphate 0.1 N: Dissolve 39 g Fe(NH
4
)
2
(SO
4
)
2
.6H
2
O in distilled water.
Add 20 mL conc. H
2
SO
4
and cool and dilute to 1 litre. Standardise this against the standard dichromate
solution. Dilute 10 mL standard K
2
Cr
2
O
7
solution to about 100 mL. Add 30 mL conc. H
2
SO
4
and cool.
Titrate with ferrous ammonium sulphate titrant using 23 drops of ferroin indicator.
2 2 7
4 2 4 2
mL K Cr O 0.25
Normality =
mL Fe (NH ) (SO )
Ammonia N
55. Zinc sulphate solution: Dissolve 100 g ZnSO
4
.7H
2
O and dilute to 1 litre.
56. EDTA reagent (stabiliser): Dissolve 50 g EDTA disodium salt in 60 mL of water containing 10 g
NaOH.
57. Nesslers reagent: Dissolve 100 g HgI
2
and 70 g KI in a small quantity of water and add this mixture
slowly with stirring to a cool solution of 160 g NaOH in 500 mL water. Dilute to 1 litre and store in
rubber stoppered pyrex glass out of sunlight.
58. Stock ammonia solution: Dissolve 3.811 g anhydrous NH
4
Cl dried at 100C in water and dilute to
1 litre. 1 mL = 1.00 mg N and 1.22 mg NH
3
.
Nitrate N
59. Stock nitrate solution: Dissolve 721.8 mg anhydrous potassium nitrate and dilute to 1 litre with distilled
water. 1 mL = 0.1 mg N.
60. Standard nitrate solution: Dilute 10 mL stock nitrate solution to 1 litre. 1 mL = 1 g N
61. Sodium arsenite solution: Dissolve 5.0 g NaAsO
2
and dilute to 1 litre.
62. Brucine-sulphanilic acid solution: Dissolve 1 g brucine sulphate and 0.1 g sulphanilic acid in about
70 mL of hot distilled water. Add 3 mL conc. HCl, cool and make up to 100 mL. This is stable for
several months.
63. Sulphuric acid solution: Carefully add 500 mL conc. H
2
SO
4
to 125 mL distilled water and cool to
room temperature.
64. Sodium chloride solution: Dissolve 300 g NaCl and dilute to 1litre with distilled water.
Nitrite N
65. Sulphanilamide reagent: Dissolve 5 g sulphanilamide in a mixture of 50 mL conc. HCl and about
300 mL distilled water. Dilute to 500 mL with distilled water.
66. N-(1-naphthyl)-ethylenediamine dihydrochloride solution: Dissolve 500 mg dihydrochloride in
500 mL distilled water. Store in a dark bottle.
67. Hydrochloric acid: HCl (1+3)
68. Stock nitrite solution: Dissolve 1.232 g NaNO
2
in nitrite free water and dilute to 1 litre. Fresh nitrite
from bottle should be taken 1 mL = 250 mg N in the solution. Preserve with 1 mL chloroform.
69. Standard nitrite solution: Standardise stock solution. Pipette 50 ml standard 0.05 N KMnO
4
, 5 mL
conc.H
2
SO
4
and 50 mL stock nitrite solution in a glass stoppered flask. Discharge the permanganate
colour by ferrous ammonium sulphate solution of 0.05 N (19.607 g ferrous ammonium sulphate and
20 mL conc.H
2
SO
4
in 1 litre) strength. Carry nitrite free blank through the entire procedure and make
necessary corrections. Calculate the nitrite N content of stock solution by the following equation:
A = [(B C) (D E)] 7/F
where, A = mg/mL nitrite N in stock solution,
B = total mL standard KMnO
4
used,
C = normality of KMnO
4
solution,
D = total mL of standard Fe(NH
4
)
2
(SO
4
)
2
used,
E = normality of standard Fe(NH
4
)
2
(SO
4
)
2
,
F = mL of stock NaNO
2
solution taken for titration.
Each 1 mL of 0.05 N KMnO
4
consumed by the nitrite corresponds to 1.729 g NaNO
2
or 350 g N.
Organic Nitrogen (to find Kjeldahl Nitrogen)
70. Digestion reagent: Dissolve 134 g K
2
SO
4
in 650 mL ammonia free distilled water and 200 mL
conc.H
2
SO
4
. Add with stirring a solution prepared by dissolving 2 g red mercuric oxide (HgO) in 25 mL
6N H
2
SO
4
. Dilute the combined solution to 1 litre.
71. Sodium hydroxide-sodium thiosulphate reagent: Dissolve 500 g NaOH and 2 g Na
2
S
2
O
3
.5H
2
O
in ammonia free distilled water and dilute to 1 litre.
72. Borate buffer solution: Add 88 mL 0.1N NaOH solution to 500 mL 0.025 M sodium tetraborate
(Na
2
B
4
O
7
) solution (5 g Na
2
B
4
O
7
in 1 litre) and dilute to 1 litre.
73. Sodium hydroxide 6 N: Dissolve 240 g NaOH in 1 litre ammonia free distilled water.
74. Standard iodine 0.1 N: Dissolve 40 g KI in 25 ml distilled water, add 13 g resublimed iodine and
stir until dissolved. Transfer to 1 litre volumetric flask and dilute to the mark.
Appendices
Appendices
APPENDIXII. STANDARDS FOR DRINKING WATER
Requirements Acceptable Cause for rejection*
Physical
1. Turbidity (Turbidity Units) 2.5 10
2. Colour units on platinum cobalt scale 5 25
3. Taste and odour Not objectionable
Chemical
1. pH 7 to 8.5 Less than 6.5 or
greater than 9.2
2. Total solids mg/L 500 1500
3. Total Hardness (as CaCO
3
) mg/L 200 600
4. Calcium (as Ca) mg/L 75 200
5. Magnesium (as Mg) mg/L 30 150
6. Iron (as Fe) mg/L 0.1 1
7. Manganese (as Mn) mg/L 0.05 0.5
8. Copper (as Cu) mg/L 0.05 1.5
9. Zinc (as Zn) mg/L 5.0 15
10. Chlorides (as Cl) mg/L 200 1000
11. Sulphates (as SO
4
) mg/L 200 400
12. Phenolic substances (as Phenol) mg/L 0.001 0.002
13. Fluorides (as F) mg/L 1.0 2.0
14. Nitrates (as NO
3
) mg/L 45 45
Toxic substances
1. Arsenic (as As) mg/L 0.05 0.05
2. Chromium (as hexavalent) mg/L 0.05 0.05
3. Cyanides (as CN) mg/L 0.05 0.05
4. Lead (as Pb) mg/L 0.1 0.1
5. Selenium (as Se) mg/L 0.01 0.01
Ratio activity
1. Alpha emitters, c/mL 10
9
10
9
2. Beta emitters, c/mL 10
8
10
8
Bacteriological quality
1. MPN index of coliform bacteria Should be zero or less 10 per 100 mL **
than 1
* Figures in excess of the permissive while not acceptable may still be tolerated in the absence of alternative and better sources, but
up to the limits designated, above which the supply will not be acceptable.
** Occasionally, the samples may show an MPN index 3 to 10 per 100 mL provided this does not occur in consecutive samples. When
consecutive samples show an MPN index exceeding 8 per 100 mL additional samples should be collected promptly from the sampling
point and examined without delay. This should be done daily until the MPN index of samples collected on two successive days is
within the acceptable limits. If necessary, samples should also be taken from several other points such as the service reservoirs,
distribution systems pumping stations and treatment plant and examined for coliforms. In addition, the operation of all treatment
process should be checked and remedial measures taken if necessary. When the results obtained over a period of one month are
considered, not more than 10% of the samples examined during the period should have shown an MPN index of coliforms greater
than 1 per 100 mL.
APPENDIXIII. MPN TABLE
No. of tubes giving positive reaction out of MPN index 95% confidence limits
3 of 10 mL 3 of 1mL 3 of 0.1 mL per 100 mL Lower Upper
each each each
0 0 0 <1
0 0 1 3 <0.5 9
0 1 0 3 <0.5 12
1 0 0 4 <0.5 20
1 0 1 7 1.0 21
1 1 0 7 1.0 23
1 1 1 11 3.0 36
1 1 0 11 3.0 36
2 0 0 9 1.0 36
2 0 1 14 3.0 37
2 1 0 15 3.0 44
2 1 1 20 7.0 82
2 2 0 21 4.0 47
2 2 1 28 10.0 150
3 0 0 23 4.0 120
3 0 1 39 7.0 130
3 0 2 64 15.0 380
3 1 0 43 7.0 210
3 1 1 75 14.0 230
3 1 2 120 30.0 380
3 2 0 93 15.0 380
3 2 1 150 30.0 440
3 2 2 210 35.0 470
3 3 0 240 36.0 1300
3 3 1 460 71.0 2400
3 3 2 1100 150.0 4800
3 3 3 >2400
1. If instead of portions of 10, 1 and 0.1 mL, a combination of 100,10 and 1 mL is used then the MPN is recorded as 0.1 times the value
given in the table.
2. For 1, 0.1 and 0.01 combination then 10 times the value in the table should be used.
3. For 0.1, 0.01 and 0.001 combination, 100 times the value given in the table should be used.
Appendices

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C. Do not allow samples to freeze.

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