EXPERIMENT FILE (ENE-252)

 EXPERIMENT – 1 : Determination of TS , TDS and TSS in Water

AIM - To determine Total Solids (TS), Total Dissolved Solids (TDS), Total
Suspended Solids (TSS), Volatile Suspended Solids (VSS) and Fixed Suspended
Solids (FSS) in the given water sample.

APPARATUS REQUIRED - Weighing scale , Petri dish , Ashless filter papers ,

Membrane filter holder , Beaker , vacuum pump , pipette , etc.

THEORY –

Water contain different types of impurities like suspended, colloidal and

dissolved materials in it. Based on its contaminants water can be broadly
defined as mixture of different types of solids in a liquid base. Solids may be
classified as settleable, suspended, dissolved, volatile and fixed. All these types
together called as total solids.

Solids form an important component in treatment processes. The suspended

and settleable solids have to be removed in the sedimentation units and the
dissolved solids will have to be removed in the filtration and other secondary
treatment facilities. The volatile solids play an important role in biological
treatment of waste water.

 Total Dissolved Solid (TDS) is the amount of combined contents of all

inorganic and organic substances contained in a liquid in molecular, ionized or
micro-granular (colloidal sol) suspended form. Total dissolved solids are
usually discussed only for freshwater systems, as salinity includes some of the
ions constituting the definition of TDS.
 Total Suspended Solids (TSS) are solids that can be trapped using a filter in
water. TSS can include a wide variation in material, such as silt, decaying plant
and animal matter, industrial wastes, and sewage. High concentrations of
suspended solids can cause several problems to stream health and aquatic life.
 Total Solids (TS) can be found by summation of TSS and TDS.
 Volatile Suspended Solids (VSS) are the suspended solids associated with
volatile fraction.
 Fixed Suspended Solids (FSS) are the suspended solids associated with the
mineral fraction.
Relevant Indian Standard for TS, TDS and TSS Test :

1. IS 3025 (Part 15)- 1984: Method of Sampling and Test (Physical and
Chemical) for Water and Wastewater : Total Residue (total Solids,
Dissolved and Suspended, First Revision.

PROCEDURE

These procedure steps will be followed on the simulator

1. When you click on total solids in water file, a new window will open .
Click on the NEXT button at the bottom right corner to move to the next
step.
2. Click on graduated pipette, now click on lower and upper arrows shown
to take sample in beaker.

3. Click on crucible to place it in weighing machine, then petri dish and

filter paper to note down the empty weights of each.
4. Click on filter paper to place it in the membrane filter and click on beaker
to pour 50ml of sample from top of filter slowly.
5. Click on pipe end to connect the filter to vacuum pump.

6. Click on filter paper to take out the filter paper from the filter and click
on temperature set button to set oven temperature at 103°C over night.
7. Click on crucible to collect the sample that passes through the filter
paper.
8. Click to oven door, click on sample to keep it in oven and adjust the
temperature set button to to set oven temperature to 115°C over night.

9. Click on Petri dish and crucible to take out from the hot air oven and
allow it cool in desiccator.
10.Click on Petri dish and crucible to take it out from the desiccator after it
cools down, click on NEXT button.
11.Note down the final weight of the ash less filter paper and final weight
of the crucible.
 OBSERVATIONS & CALCULATIONS (1)

12.Calculate the values of total suspended solids, total dissolved solids and
total solids then enter the value and click on CHECK to check the results,
then proceed with the further process.

13.Ignite the filter paper with the solids in a muffle pot at 600°C in a muffle
furnace.
14.Take out the muffle pot from the desiccator and then note down the
weight of it.
 OBSERVATIONS & CALCULATIONS (2)

15.Now , calculate the values of fixed suspended solids and volatile

suspended solids and then check the results.

 EXPERIMENT . 2 : Determination of Alkalinity in Water .

AIM : To determine acid neutralizing capacity of given water sample.
APPARATUS REQUIRED : Burette , pipette , pipette bulb , Conical Flask , 250 mL
measuring cylinders , Standard Flask , beakers , etc.

THEORY :

Alkalinity is the presence of sufficient alkaline ions in water. The absence of

alkalinity makes the water acidic. Water is said to be alkaline when pH of water is
above 7.00. The determination of alkalinity is very useful in water and waste water
because it provides buffering to resist changes in pH value. The alkalinity and acidity
in water usually changes in natural waters with large algal growth. The alkalinity of
natural waters is primarily due to the salts of week acids and bicarbonates represent
the major form of alkalinity. Alkalinity can be expressed as follows:

Alkalinity (mol/l) = [HCO3-] + 2[CO3 2-] + [OH-] – [H+]

The presence of alkalinity in surface waters is primarily indicates the

concentration of carbonate, bicarbonate and hydroxide contents. Alkalinity in excess
of Alkaline earth metal concentrations is significant in determining the suitability of a
water for irrigation. Alkalinity measurements are used in the interpretation and
control of water and waste water treatment processes. Raw domestic waste water
has an alkalinity less than or only slightly greater than that of the water supply.

Alkalinity of water is the capacity of water to neutralize acid. It is usually

expressed as Total Alkalinity or Caustic Alkalinity in water. It is significant in many
uses and treatments of natural waters and wastewaters. Alkalinity is measured
titrimetrically by titrating against Dilute Sulphuric Acid. Phenolphthalein and Methyl
Orange are used as indicator to indicate pH 8.3 and pH 4.3. Phenolphthalein
process a pink colour when pH is above 8.3 and colourless when pH is below 8.3.

Relevant Indian Standard for Alkalinity Test :

1. IS 3025-23 (Part 23) - 1986: Methods of Sampling and Test (physical and
chemical) for Water and Wastewater, Alkalinity, First Revision.

PROCEDURE

These procedure steps will be followed on the simulator

1. Open the simulation of alkalinity of water and click on the arrow mark shown
at the bottom right corner.
2. Click on the funnel to place it on the burette.
3. Click on the lid to open and click on the bottle to pour dilute sulphuric acid on
to the burette up to zero mark.
4. Remove the funnel by clicking on it. Click on the NEXT button once the funnel
is removed.
5. Squeeze the pipette bulb by clicking on it and press the up arrow to take the
Na2CO3 solution up into the pipette.

6. Press the down arrow on the pipette bulb to release liquid into the conical
flask.
7. Note the addition of 10ml 0.02N Sodium carbonate solution to conical flask.
Click on NEXT button.
8. Click on the dropper to add methyl orange indicator to the conical flask.
9. After the addition observe the change in colour of the solution, then click on
the NEXT button.
10. Click on the knob to add the dilute sulphuric acid into the conical flask.
11. Observe the colour change of the solution in the conical flask to wine red.
Click on the NEXT button.
 OBSERVATIONS & CALCULATIONS :

FORMULA USED :
 N1V1=N2V2 , where N1= Normality of H2SO4 , V1= Volume of H2SO4 , N2=
Normality of Na2CO3 , V2= Volume of Na2CO3
 T as Alkalinity of CaCO3 = Nacid x Vacid x 5000 / Vsample

TABLE 1 : TOTAL ALKALINITY:

S.NO Volume of Initial Final burette Volume of

sample (in burette reading Sulphuric
mL) reading Acid (mL)

1.

2.

3.

PRECAUTIONS :
1. Do not keep the indicator solution open since it contains the alcohol
which tends to evaporate.
2. The mixed indicator solution contains dye in it ; Care should be taken
so that it is not spilled to your skin.
  EXPERIMENT 03 : Determination of Hardness in Water

To determine the amount of Total Hardness present in a given water sample

by EDTA Titration method.

INTRODUCTION

Hardness is a degree of ability of water to cause precipitation of

insoluble calcium and magnesium salts of higher fatty acids from soap
solutions. Hardness can be categorized into two types– permanent hardness
and temporary hardness. Temporary hardness is caused due to the
presence of bicarbonates of calcium and magnesium. Permanent hardness
is non-carbonate hardness and is caused due to the presence of sulphates,
chlorides and nitrates of calcium and magnesium.

Hardness of water is significant in determining the suitability of water

for domestic and industrial uses. The comparative amount of calcium and
magnesium hardness, carbonates and non-carbonates hardness present in
water are the aspects while determining the most economical type of
softening process.

The ions involved in water hardness, i.e. Ca 2+(aq) and Mg2+(aq), can be
determined by titration with a chelating agent, Ethylene Diamine Tetra
Acetic acid (EDTA), usually in the form of disodium salt (H2Y2-).

The titration reaction is: Ca2+(aq) + H2Y2-(aq) → CaY2-(aq) + 2H+(aq)

Eriochrome Black T is often used as indicator for the above titration.

At pH 10, Ca2+(aq) ion first complexes with the indicator as CaIn +(aq) which is
wine red. As the stronger ligand EDTA is added, the CaIn +(aq) complex is
replaced by the CaY2-(aq) complex which is blue. The end point of titration is
indicated by a sharp colour change from wine red to blue. Titration using
Eriochrome Black T as indicator states total hardness due to Ca 2+(aq) and
Mg2+(aq) ions. Hardness due to Ca 2+(aq) ion is determined by a discrete
titration at a higher pH, by adding NaOH solution to precipitate Mg(OH) 2(s),
by means of hydroxy naphthol blue as indicator.

Relevant Indian Standard for Hardness Test on Water :

1. 1. IS 3025 (Part 21) -2009: Method of Sampling and Test (Physical and
Chemical) for Water and Wastewater, Hardness, Second Revision.

these procedure steps will be followed on the simulator

1. Open the simulation of hardness of water, go through the given

Description and Solutions used and click on NEXT button shown at the
bottom right corner.
2. Click on funnel to place it in the burette to add 200ml test sample.
3. Click on the lid to open and pour EDTA solution on to the burette upto
zero mark.
4. Remove the funnel by clicking on it.
5. Click on NEXT button once the funnel is removed.
6. Squeeze the pipette bulb by clicking on it and press the up arrow to
take the CaCO3 solution up into the pipette.
7. Press the down arrow to release liquid into the beaker.
8. Note the addition of 10ml 0.01M Calcium carbonate solution to
conical flask. Click on NEXT button.
9. Click on the pipette to add ammonia buffer solution to the conical
flask.
10. Select answer to the displayed question and then click on the NEXT
button.
11. Click on the dropper to add 5-6 drops of E. B.T indicator to the conical
flask.
12. Observe the colour change of solution to wine red and click on the
NEXT button.
13. Click on the knob to add the ETDA solution into the conical flask.
14. Calculate CaCO3 equivalent to 1ml of ETDA using the formula, enter
the value and click on CHECK.
 15. Repeat the same procedure for the 100ml of water sample. Calculate
the total hardness of water using the formula and enter the value and
click on CHECK.
FORMULA USED

 EXPERIMENT 04 :Determination of Chloride in Water

AIM-To determine Chloride Ion Concentration in a water sample by Mohr's

method.

INTRODUCTION

Chloride, a naturally occurring element found most commonly in

natural waters. It is often found as a component of salt (sodium chloride) or
in combination with potassium or calcium. The weathering of soils, salt-
bearing geological formations, salt used for road de-icing, contributions
from wastewaters and in coastal areas, intrusion of salty ocean water into
fresh groundwater are the main sources of chloride in water. Chlorides can
corrode metals and affect the taste of food products. It is not usually
harmful to humans; however, the sodium part of table salt has been linked
to heart and kidney disease. Chloride is basically fully absorbed in normal
individuals until the content is too high. Presence of Sodium chloride in
water gives salty taste at 250mg/l, calcium or magnesium chloride does not
impart any taste until it reaches 1000mg/l.

The measured chloride ions can be used to know salinity of different

water sources. For brackish water (or sea water or industrial brine solution),
it is an important parameter and indicates the extent of desalting of
apparatus required. It also interferes with COD determination and thus it
requires a correction to be made on the basis of amount present or else a
complexing agent, such as HgSO4 can be added. Further, chloride ions are
used as tracer ions in column studies to model fate of different
contaminants in soil and liquid media.

Effects on humans :

A normal adult human body contains approximately 81.7g chloride.

Based on a total obligatory loss of chloride of approximately 530mg/day, a
dietary intake for adults of 9mg of chloride per kg of body weight has been
recommended (equivalent to slightly more than 1g of table salt per person
per day). For children up to 18 years of age, a daily dietary intake of 45mg of
chloride should be sufficient. A dose of 1g of sodium chloride per kg of body
weight was reported to have been lethal in a 9-week-old child. Little is
known about the effect of prolonged intake of large amounts of chloride in
the diet.

Principal involved :

The Mohr's Method uses silver nitrate for titration (normality: 0.0141)
(method applicability: 0.15 to 10 mg/l chloride ions). This corresponds to
1ml of 0.0141 equals to 1mg chloride in solution. The silver nitrate solution
is standardized against standard chloride solution, prepared from sodium
chloride(NaCl). During the titration, chloride ion is precipitated as white
silver chloride.

(Eg.1) : Ag+ + Cl- ⇔ AgCl (Solubility product constant, Ksp=3×10-10)

The indicator (potassium chromate) is added to visualize the

endpoint, demonstrating presence of excess silver ions. In the presence of
excess silver ions, solubility product of silver chromate exceeded and it
forms a reddish-brown precipitate (Eq.2). This stage is taken as evidence
that all chloride ions have been consumed and only excess silver ions have
reacted with chromate ions:

(Eg.2) : 2Ag+ + CrO42- ⇔ Ag2CrO4 (Ksp=5×10-12)

Relevant Indian Standard for Chloride in Water Test :

1. IS 3025 (Part 32) -1988: Method of Sampling and Test (Physical and
Chemical) for Water and Wastewater- Chlorides, First Revision.
These procedure steps will be followed on the simulator

1. When you click on a Chloride in water file, a new window will open as
shown below, go through the Description and Solutions used, then
click on the next button at the bottom right corner to move to the
next step.
2. Click on funnel to place it in the burette and click again to silver
nitrate (AgNO3) solution bottle to add it to the burette.
3. Click on pipette to add 10ml of 0.0141N standard sodium chloride
(NaCl) solution to a conical flask carefully without forming air bubbles.
4. Click on the dropper to add 10 drops of potassium chromate as
indicator to the conical flask.
5. Click on the knob to start titrating sodium chloride solution till the
colour change from yellow to pinkish yellow.
6. Stop titration when the colour change is seen from yellow to pinkish
red and note down the initial and final burette reading.
7. Click on the formula text box and calculate the Normality of silver
nitrate solution, then click on check to evaluate the result.
8. Obtain the normality of silver nitrate solution and carry out the
sample procedure of titration for blank test using distilled water
sample and sample test using water sample.
9. Display the calculation and obtain the value of chloride in water.
 EXPERIMENT-5
AIM: To determine concentration of Total Iron content in water sample by
Spectrophotometric method.

THEORY: The percolation of water through soil and rock dissolves minerals like Iron
and Manganese in that and takes along with its flow. Water containing iron and manganese
is actually colourless when it is not in contact with the oxygen. But when the same water is
exposed to air, it gets oxidized causing change in water colour. The oxidation of dissolved
iron particles in water changes the iron to white, then to yellow and finally to red-brown
solid particles that settle out of the water. Iron that does not form particles large enough to
settle out and that remains as colloidal iron leaves the water with a red tint. Although
manganese is present in water in a dissolved form it leaves black tint which is due to the
colloidal manganese in shallow wells. This water containing Iron is one of the minerals
required by the human body. It is used in the production of the oxygen-carrying proteins,
haemoglobin and myoglobin. A deficit of iron in the body can leave a person feeling tired
and listless, and can lead to a disorder called anemia. Presence of iron in water is not
harmful for health, but it renders the water turbid, imparts colour to water and hence
unacceptable from the aesthetic point. It causes satins in laundries and plumbing fixtures.
Its presence may help in the proliferation of iron reducing bacteria in the aquifer and the
pipe lines. Hence, the desirable limit is 0.3mg/l in water and the tolerable limit is 1.0mg/l.

To detect the presence of these ions in solution, Thiocyanate ions (SCN -) are added.
Thiocyanate ions respond with iron (III) ions in solution to form a deep red coloured
complex ion:

Fe3+(aq) + SCN-(aq) → [FeSCN]2+(aq)

or, more fully, [Fe(H2O)6]3+(aq) + SCN-(aq) → [Fe(H2O)5SCN]2+(aq) + H2O (l)

By comparing the intensity of the colour of this solution with the colours of a series
of standard solutions, with known Fe3+ concentrations, the concentration of iron in the food
sample may be determined. This method is called colorimetry. Colorimetric analysis is based
on the change in the intensity of the color of a solution with dissimilarities in concentration.
Colorimetric methods represent the simplest form of absorption analysis. The human eye is
used to associate the color of the sample solution with a set of standards until a match is
found. An increase in sensitivity and accuracy results when a spectrophotometer is used to
measure the color intensity. Mainly, it measures the fraction of an incident beam of light,
which is transmitted by a sample at a particular wavelength.

Relevant Indian Standard for Total Iron Test on Water:

1. IS 3025 (Part 53) -2003: Method of Sampling and Test (Physical and Chemical) for
Water and Wastewater- Iron, First Revision.
 PROCEDURE

To obtain the following quantity of chemicals from the back bench using a labeled beakers.
Unknown iron solution ~15 mL Standard iron solution (0.2500 g/L) ~30 mL 0.3%
weight/volume o-phenanthroline solution ~40 mL 10% weight/volume hydroxylamine
hydrochloride ~40 mL

2. Prepare the following iron calibration solutions by pipetting the indicated amounts of the
above iron solution (step 1) into labeled 50 mL volumetric flasks. The first flask is a blank
containing no iron. Concentration of Fe Volume to pipet 0.00 mg Fe 0.00 mL 0.05 mg Fe 4.00
mL 0.10 mg Fe 8.00 mL 0.15 mg Fe 12.00 mL 0.20 mg Fe 16.00 mL 0.25 mg Fe 20.00 mL

3. Pipet 10.00 mL of an unknown sample solution (record the unknown’s number) into a 250
mL volumetric flask and dilute to the mark with distilled water. Invert and shake the flask
several times to mix the solution.

4. Pipet two 25.00 mL aliquots of this solution into two 50 mL volumetric flasks labeled
unknown.

5. Using a 10 mL graduated cylinder, add 4.0 mL of 10% hydroxylamine hydrochloride

solution and 4.0 mL of 0.3% o-phenanthroline solution to each volumetric flask.

6. Swirl and allow the mixture to stand for 10 minutes.

7. Dilute each flask to the mark with distilled water and mix well by inverting and shaking
the capped volumetric flasks several times.

8. Using the Spectronic 301 spectrophotometer, carefully measure the percent

transmittance of the various solutions in the 50 mL volumetric flasks, including the two
unknown solutions. Record your results in the following table. Solution % Transmittance
Absorbance (A=-logT) 0.00 mg Fe (blank) 100% 0 0.05 mg Fe 0.10 mg Fe 0.15 mg Fe 0.20 mg
Fe 0.25 mg Fe

Calculations and Discussion

1. Prepare a plot of absorbance versus concentration of the known solutions (express the
concentration in mg Fe per 50 mL of solution). Draw the best fitting straight line through the
points – this is called the Beer-Lambert Law plot.

2. Place the best Absorbance value of each unknown solution onto this plot and determine
their concentrations.

3. Calculate the amount of iron in the unknown sample. Express this as mg of Fe per litre of
the original unknown solution (mg/L Fe). E.g. From the graph you obtain a concentration of
0.10 mg Fe/50 mL Since in step 3 we diluted the original sample 25 times and in step 4, 2
more times the concentration of the original sample is therefore: L mg Fe L mL dilution
factor mL mg Fe 1000 100 50( ) 50 0.10 ´ ´ =

4. Compare your results with the accepted value

Unknown #1 Unknown#2 Unknown#3 Unknown #4

173.5 mg/L 209.2 mg/L 225.6 mg/L 242.7 mg/L

And

calculate the relative error= [ (experimental value- accepted value )/ accepted value ]
*100%.

EXPERIMENT-6
 Aim: To determine the pH of the given sample.
 Theory: pH of aqueous solutions can be defined as negative logarithm of hydrogen
ion concentration. pH values ranging from 0 to 7 are acidic, and from 7 to 14 are
alkaline. The effect of pH on the chemical and biological properties of liquids makes
its determination very important. It is used in several calculations in analytical work.
The pH determination is usually done by electrometric method which is the most
accurate method and free of interference.
 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 sludge, 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, CO2 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.
 Standard: pH of drinking water should be between 6.5 and 8.5
 Requirements: pH Meter, Flask, Magnetic Stirrer, Funnel, Beaker, Wash Bottle
 Electrometric Method: The pH is determined by measurement of the electromotive
force of a cell comprising an indicator electrode (an electrode responsive to
hydrogen ions such as glass electrode) immersed in the test solution and a reference
electrode or a combined electrode. The contact between the test solution and the
reference electrode is usually achieved by means of a liquid junction. The emf of this
cell is measured with pH meter. This is a high impedance electrometer calibrated in
terms of pH. Reagents: Calibrates the electrode system against standard buffer
solution of known pH. Buffer tablets having pH 4.0, 7.0 and 9.2 are available.
 Reagents: pH 4, pH7, pH 9.2 Buffer Solutions: Dissolve buffer tablet of pH 4 in 100 ml
of distilled water to get pH 4 buffer solution. Similarly other buffer solutions can be
prepared by dissolving corresponding buffer tablets.

PROCEDURE

1. Prepare buffers of pH 4, 7 and 9.2.

2. Rinse the electrode thoroughly with distilled water.
3. Standardize pH meter electrode with any two of these buffers.
4. Dip the electrode in the sample for test.
5. Let the reading on screen stabilize; note it as pH & temperature of
your sample.
 EXPERIMENT 07

Aim: To find the Biological Oxygen Demand exerted by the given sample of waste water.

Theory:
The Biochemical Oxygen Demand (B.O.D.) of sewage or of polluted water is the amount of
oxygen required for the biological decomposition of dissolved organic matter to occur under
aerobic condition and at the standardised time and temperature. Usually, the time is taken
as 5 days and the temperature 20°C as per the global standard. The B.O.D. test is among the
most important method in sanitary analysis to determine the polluting power, or strength of
sewage, industrial wastes or polluted water. It serves as a measure of the amount of clean
diluting water required for the successful disposal of sewage by dilution. The test has its
widest application in measuring waste loading to treatment plants and in evaluating the
efficiency of such treatment systems.

The test consists in taking the given sample in suitable concentrations in dilute water in
B.O.D. bottles. Two bottles are taken for each concentration and three concentrations are
used for each sample. One set of bottles is incubated in a B.O.D. incubator for 5 days at
20°C; the dissolved oxygen (initial) content (D1) in the other set of bottles will be determined
immediately. At the end of 5 days, the dissolved oxygen content (D2) in the incubated set of
bottles is determined. Then,

Where,

P = Decimal fraction of sample used.

D1 = Dissolved oxygen of diluted sample (mg/l), immediately after preparation.

D2 = Dissolved oxygen of diluted sample (mg/l), at the end of 5 days incubation.

Among the three values of B.O.D. obtained for a sample select that dilution showing the
residual dissolved oxygen of at least 1mg/l and a depletion of at least 2mg/l. If two or more
dilutions are showing the same condition then select the B.O.D. value obtained by that
dilution in which the maximum dissolved oxygen depletion is obtained. Waste water from
sewage treatment plants often contains organic materials that are decomposed by
microorganisms, which use oxygen in the process. (The amount of oxygen consumed by
these organisms in breaking down the waste is known as the Biochemical Oxygen Demand
or BOD).

Objectives that we accomplish from BOD determination :

 To assess the quality of surface water and waste water

 To establish the amount of organic matter in samples

 To understand the significance of Dissolved Oxygen in sample

Relevant Indian Standard for Biological Oxygen Demand Test :

1. IS 3025 (Part 44)-1993: Method of Sampling and Test (Physical and Chemical) for
Water and Wastewater, Biochemical Oxygen demand, First Revision.

Procedure
These procedure steps will be followed on the simulator

1. When you click on a BOD file, a new window will open . Go through the solutions and
description, then click on the NEXT button at the bottom right corner to move to the
next step.

2. Click on the arrow mark and measure the Test Sample to be taken.
3. The measured 200ml Test Sample is taken in two BOD Bottles.

4. Dilute two BOD bottles with 200ml Test Sample by filling distilled water up to the
bottle neck and label it as Sample A and Sample B. Select answer to the question
displayed and then click Next.
5. Click on automatic pipette to add 1ml of MnSO4 solution to Sample A bottle.

6. Add 1ml of Alkali-iodide-azide solution to Sample A bottle.

7. Again follow the same procedure to add Alkali-iodide-azide solution to Sample A

bottle carefully so that there is no bubble formation. Click Next.
8. Mix the solution by shaking BOD bottle upside-down for 25-30 times and allow the
precipitate to settle down at bottom.

9. Click on arrow pointer to squeeze the pipette bulb and dip into the H 2SO4 solution.

10. Add 1-2ml of conc. H2SO4 carefully without forming air bubbles. Press the up and
down arrow on the bulb to take and release the liquid into the BOD bottle.

11. Click on the bottle to shake it by inverting the BOD bottle till all the precipitate
dissolves.

12. Take 200ml of sample in a conical flask using pipette.

13. Click on upper and lower arrow to take sample and add it to the flask.

14. Click on funnel to place it in the burette and the burette knob to take Na 2S2O3 in the
burette.

15. Titrate the solution against 0.025N Na2S2O3. Click on knob to start the liquid running
into the conical flask.

16. Add 1ml Starch once the colour of the solution changes to pale yellow, now the
colour changes to blue then continue the titration till blue colour becomes
colourless.
17. Note down the initial and final burette reading. Click Next.

18. Keep Sample B and put it into the BOD incubator for 5 days. Calibrate the
temperature of BOD incubator for approximately 20°C.

19. Set the temperature of incubator to 20°C and wait for 5 days.

20. Repeat step 5 to 16 for Sample B. Note down the Initial and final Burette reading.
Determine the BOD of the sample using formula.
 Experiment-8

AIM : To determine the Chemical Oxygen Demand of the given water sample.
APPARATUS : COD digester , Round bottom reflux flask , Digestion vessels ,
Conical flask , Burette , Pipette , etc.

THEORY : The Chemical Oxygen Demand is a measurement of the amount of

material that can be oxidized in the presence of a strong chemical oxidizing
agent.Other related analytical values are Total Oxygen Demand (TOD),
Biochemical Oxygen Demand (BOD), total organic carbon (TOC).

The COD value will always be higher than the value of BOD for wastewater.

COD = 1.6 BOD5

In COD, a strong oxidizing agent such as potassium dichromate in acid medium

is used for chemical oxidation of organic matter. It is measured in mg/l or ppm.
Chemical Oxygen Demand is an important water quality parameter because,
similar to BOD, it provides an index to assess the effect discharged wastewater
will have on the receiving environment. The COD test is often used as an
alternative to BOD due to shorter length of testing time.

Organic matter + Cr2O72- + H+ + (acid) → Cr3+ + CO2 + H2O

Oxidant Cr6+ Cr3+

Organic Matter → CO2 + H2O

Empirically for municipal wastewater, BOD5 ≈ 0.4 to 0.6 COD. Even though
COD does not represent exactly the biodegradable organics, it is sometimes
taken as a measuring parameter of organic waste, as a COD value can be
determined in 3-4 hours as against 3-5 days for a BOD value. COD can be
determined titrimetrically or photometrically.

Relevant Indian Standard for Chemical Oxygen Demand Test :

IS 3025 (Part 44)-1993: Method of Sampling and Test (Physical and Chemical)
for Water and Wastewater, Biochemical Oxygen demand, First Revision.

PROCEDURE :
1. Take 10 ml of sample into a round bottom reflex flask.
2. Add 1 ml of Mercury sulfate (HgSO4 ) solution to the flask and mix by
swirling the flask.
3. Add 5 ml of Potassium dichromate (K2Cr2O7 ) solution.
4. Now add slowly and carefully 15 ml Silver sulfate- Sulfuric acid solution.
5. Transfer the content in the flask to the digestion vessel. Then place the
digestion vessel in the COD digester.
6. Take 0.025M Ferrous Ammonium Sulphate solution in a burette.
7. Titrate the solution till the colour changes from blue to colourless.

OBSERVATIONS & CALCULATIONS

Experiment-9
AIM : To determine the optimum Alum dosage for water or waste
water treatment by Jar test.

APPARATUS : Nephelometer , Nephelometer tube , Pipette, Beaker ,

Stirrer and Measuring cylinder , etc.

THEORY: Generally, we encounter very fine and charged clay like

particles in water treatment, which should be detached before we
continue for further processes. When water is passed through a
sedimentation tank, these impurities do not settle down by gravity. The
cause being that these are charged particles, they repel each other and
just stay.

Coagulation is the process of addition of a chemical to destabilize a

stabilized charged particle.

Flocculation is a slow mixing technique which promotes agglomeration

and helps the particles to settle down.

The presence of fine charged particles increases the turbidity of the water,
which is undesirable, and hence these impurities are to be removed.
Coagulation removes not only turbidity, but also colour, odour producing
substance, microorganisms, algae, phosphate and taste.

In the current experiment we are using "Alum" [Al 2(SO4)3 18H20] as the
clarifying agent.Molecules dissociate to yield SO42- and Al3+ when alum
solution is added to water. In order to neutralize the charge these charged
species combine with the charged colloidal particles.

Jar Test :

Jar test is a common laboratory method used to determine the

optimum operating conditions for water or wastewater treatment. This
process allows changes in pH, variations in coagulant or polymer dose,
alternating mixing speeds, or testing of different coagulant or polymer
types, on a small scale in order to predict the working of a large-scale
treatment operation.

Jar Testing Apparatus :

The jar testing apparatus consists of six paddles, which stir the contents of
six one-litre containers. One container acts as a controller while the
operating conditions can be varied among the residual five containers. An
rpm gage at the top-centre of the device allows for the constant control of
the mixing speed in all of the containers.

Relevant Indian Standard for Alum Coagulation Test:

IS 3025 (Part 50)-1986: Method of Sampling and Test (Physical and

Chemical) for Water and Wastewater: Jar Test (Coagulation Test), First
Revision.

PROCEDURE
1. The jar test procedures involves the following steps:
Fill the jar testing apparatus containers with sample
water. One container will be used as a control while the
other 5 containers can be adjusted depending on what
conditions are being tested. For example, the pH of the
jars can be adjusted or variations of coagulant dosages
can be added to determine optimum operating conditions.
2. Add the coagulant to each container and stir at
approximately 100 rpm for 1 minute. The rapid mix stage
helps to disperse the coagulant throughout each
container.
3. Turn off the mixers and allow the containers to settle for
30 to 45 minutes. Then measure the final turbidity in each
container.
4. Reduce the stirring speed to 25 to 35 rpm and continue
mixing for 15 to 20 minutes. This slower mixing speed
 helps promote floc formation by enhancing particle
collisions which lead to larger flocs.
5. Residual turbidity vs. coagulant dose is then plotted and
optimal conditions are determined. The values that are
obtained through the experiment are correlated and
adjusted in order to account for the actual treatment
system.

OBSERVATIONS
 Experiment – 10
Aim: To determine Break-Point Chlorine Demand in water sample.
Apparatus used: BOD Bottles, Pipette, Conical flask, Burette, Dropper,
BOD Incubator etc.

Theory: Chlorination of water supplies and wastewater treatment plant

effluents is mainly done for the destruction of disease-producing
microorganisms. Chlorine has many other uses in the wastewater
treatment plant operation including odour control and fly and ponding
control (in trickling filters). By reacting with ammonia, iron, manganese,
sulphide and some organic substances, chlorine can provide additional
wastewater treatment. 

 Unfortunately, if the use of chlorine is not controlled carefully it might

cause problems. When used as a control on the disease causing
bacteria, the idea is to disinfect and not sterilize the effluent. Disinfection
is the process of killing disease-producing bacteria. 

 Process of killing all living organisms is called as sterilization. If an

attempt were made to sterilize the effluent, the biological life in the
receiving waters would likely be destroyed. The best process for
monitoring the disinfection process and preventing harm to aquatic life
by over-chlorination is to maintain a specific chlorine residual level in the
effluent.

It will also increase the quality of water by reacting with ammonia, iron,
manganese, sulphide and some organic substances. The residual
chlorine is sustained in water to promote the primary purpose of
chlorination. This method of determination depends upon the oxidising
power of free and combined chlorine residuals. 

 At pH 8 or less, chlorine will release free iodine from potassium iodide
solution. The released iodine is titrated against standard sodium
thiosulphate solution using starch indicator.

2KI + Cl2 → 2KCL+I2

I2 + Starch → Blue Color

I2 + 2Na2S2O3 → Na2S4O6 + 2NaCl + 2Na

 Breakpoint chlorination is the point where the demand for chlorine
has been fully satisfied in terms of chlorine addition to the water. When
chlorine is added to water, a reaction is produced in the compounds
present in the water.

Residual chlorine is the chlorine remaining in water after contact

period (of 30 to 60 min). During chlorination process, sufficient quantity
of chlorine must be added to reach 0.01 to 0.02 mg/l of residual content
of non-reacted chlorine in treated water.

Super chlorination is a water treatment process in which the addition

of excess amounts of chlorine to a water supply speed chemical
reactions or insure disinfection within a short contact time .

Procedure
 Take 200ml of test sample in 26 bottles.
 Add standard Chlorine solution to the bottles through pipette in
0.5ml increment.
 Mix the sample by inverting the bottle and keep all the bottles in
dark for 20 minutes.
 Pour 200ml of sample from each bottles into conical flask.
  Add 5ml of concentrated Sulphuric Acid to the conical flask.
 Add 1g Potassium Iodide (KI) crystals and stir the flask properly.
 Add 1ml of Starch indicator to conical flask and observe the
change in color of the solution from colorless to very light blue.
 Take 0.0282N Iodine solution bottle to fill the burette.
 Titrate the solution against 0.0282N Iodine solution until colour
changes from blue to light pink.
 Observe the colour change from light pink to colourless.
 With the help of formula used and observations obtained,
calculate Residual Chlorine.
  Continue the titration procedure for all the remaining samples
and obtain the Residual Chlorine value.

Observations
 Chlorine Titration Residual
Dosage(mg/L) value(mL) Chlorine(mg/L)

Experiment: 11
Aim: To determine Sulphate ion concentration in sample by Turbidimetric
method.

Apparatus used: Spectrophotometer, Cuvettes, Pipette, Conical flask,

Magnetic stirrer, Spatula, etc.
Theory: Sulphates is largely distributed in nature and may be present in
natural waters in concentration ranging from few hundred to several thousand
mg/l. It occurs naturally in frequent minerals, including barite, epsomite and
gypsum. 

These dissolved minerals subsidize to the mineral content of

drinking-waters. Acid Mine Drainage (AMD) may give large amounts of
sulphates through pyrite oxidation. 

It is the second most abundant anion in seawater. Its high

concentration owes to the high to reasonable solubility of the salts that it
forms with the foremost cations in seawater, namely, Na, Mg 2+ and Ca2+.

Environmental Significance:

 Sulphates are of substantial concern because they are indirectly liable

for two serious problems frequently related with handling and
treatment of wastewater. They are odour and sewer corrosion problem
result from the reduction of sulphates to hydrogen sulphide under
anaerobic conditions.
 The amount of sulphates in wastewater is a feature of concern in
defining the magnitude of problems that can rise from reduction of
sulphates to hydrogen sulphide. 
 For instance, knowledge of the sulphates content of the sludge or waste
fed to digestion units provides a means of estimating the hydrogen
sulphide content of the gas produced. From this information, the design
engineer can determine whether scrubbing facilities will be needed to
remove hydrogen sulphide and size of the units required.

Principle:
The turbidimetric method of measuring sulphates is based upon the
fact that barium sulphates tend to precipitate in a colloidal form of uniform
size and that this tendency is enhanced in presence of a sodium chloride,
hydrochloric acid and glycerol.
SO42- + BaCl2 → BaSO4 + 2Cl-
The absorbance of the barium sulphates formed is measured by a
spectrophotometer at 420nm and the sulphates ion concentration is
determined by evaluation of the reading with a standard curve.
 Procedure
 Prepare a series of standards of 100mL by accurately pipetting
calculated volumes of standard sulphate solution to measure 5 to
40 mg/L in conical flasks.
 Dilute the solution by adding distilled water.
 Similarly fill 6 flasks with sulphate solution in 5mL increment and
dilute it with distilled water.
 Prepare a blank solution and a sample solution by adding 100mL
of distilled water and water sample to conical flask respectively.
 Add 20ml of the Buffer solution to conical flask.
 Stir the solution in conical flask using magnetic stirrer at
constant speed.
  Add 1 spatula of Barium Chloride crystals stir it for 60-65sec
and allow it to rest.
 Fill the cuvette with blank solution and place it in
spectrophotometer. Set the wavelength to 420nm by adjusting
wavelength control knob.
 Fill the cuvette with Sulphate solution and place in the
Spectrophotometer and note down absorbance and transmittance
values for all solutions.
 Note down the absorbance and transmittance values for all
solutions.

Observations:

Sampl Sulphate Absorbanc Transmittanc

e Concentration(mg/L e e
)
  Experiment – 12
Aim: To determine Fluoride concentration in the given water sample
by Ion Selective Method.

Theory: Fluoride present can be determined by either colorimetric

method or Ion Selective Method. With colorimetric method, higher
concentrations of fluoride cannot be determined. With ISE method,
wide range of fluoride can be determined. Further, color and
turbidity will not interfere.
The fluoride electrode is an ion-selective sensor. The key element in
the fluoride electrode is the laser-type doped lanthanum fluoride
crystal across which a potential is established by fluoride solutions of
different concentrations. The crystal contacts the sample solution at
one face and an internal reference solution at the other. The
reference electrode used is Ag/AgCl.
Fluoride forms complexes with several polyvalent cations, notably
aluminium and iron. Use of TISAB provides uniform ionic strength
background, adjusts pH and breaks up such complexes. CDTA
(Cyclohexylene Diaxnine Tetra Acetic Acid), a component present in
TISAB (Total Ionic Strength Adjusting Buffer), preferentially will
complex with interfering cations and release free fluoride ions. By
maintaining pH above 5.0, complex formation of hydrogen fluoride is
avoided and interference of hydroxide ion is avoided by maintaining
the pH below 5.5.

Relevant Indian Standard for Fluoride Test:

1. IS 3025 (Part 60) -2008: Method of Sampling and Test (Physical

and Chemical) for Water and Wastewater: Fluoride, First
Revision.

Apparatus used: Ion Selective Electrode (ISE) meter, Fluoride Ion

Selective Electrode, Magnetic stirrer with TFE-coated stirring bracket,
Beaker, Pipette.

PROCEDURE

1. Pipette 2.5 ml, 5.0 ml, and 10.0 ml of standard Fluoride solution
in a beaker.
2. Dilute the solution by adding distilled water up to 100ml mark.
3. Use the graduated pipette to add 100 ml of sample water to
prepare a sample solution.
4. Add 25 ml of TISAB solution to the sample solution.
5. Clean the electrodes using distilled water, then dry the Fluoride
ISE electrodes using tissue paper.
6. Place the 2.5 ml Fluoride solution on a stirring plate and add a
magnetic stirring bar. Then dip the Fluoride ISE electrodes in
the solution. Then repeat the same for 5.0 and 10.0 ml Fluoride
solution.
7. Place the water sample on the stirring plate and add a magnetic
stirring bar. Then dip the Fluoride ISE electrodes in the solution
to determine the Fluoride concentration of it.
 Observations: Concentration of Fluoride in the given water
sample
= 0.12 mg/l

 Inference: Permissible limit of Fluoride in water is < 1mg/l. The

water sample given has Fluoride level = 0.12mg/l, so it is
suitable for drinking.
 EXPERIMENT - 13

Determination of Dissolved Oxygen in Water

AIM:
To estimate Dissolved Oxygen (DO) level in the given water sample
by Winkler's Method

THEORY:

INTRODUCTION

The major inputs of Dissolved Oxygen (DO) to natural water are

from atmosphere and photosynthetic reaction. Where the algae and
phytoplankton production is high, the saturation of oxygen can occur
during daytime. The solubility of oxygen in water depends on
temperature and other factors. The presence of oxygen is essential
for the survival of aquatic life in water. A rapid fall of DO in river water
is one of the first indications of pollution.

Significance :
 It is necessary to know the DO level to assess quality of raw water and to keep a check on stream
pollution.

 A minimum DO of 4 to 5 mg/l is desirable for the survival of aquatic life.

 Higher values of DO may cause corrosion of Iron and steel.

 DO test is used to evaluate the pollution strength of domestic and industrial wastes.

Winkler’s Modified Method :

The Winkler’s Method is the technique used to measure DO in
freshwater structures. It is used as an indicator of the health of a
water body, where higher DO concentrations are associated with high
production and little contamination. This test is performed on-site, as
delays between sample collections and testing may result in a
variation in oxygen content.
 In this method of analysis the oxygen present in the water
sample oxidizes the divalent manganous ion to its higher valency,
which precipitates as a brown hydrated oxide after addition of sodium
hydroxide (NaOH) and potassium iodide (KI). Upon acidification,
manganese reverts to divalent state and liberates iodine from KI
equivalent to DO content in the sample. The liberated iodine is titrated
against sodium thiosulphate (0.025N), using freshly prepared 2%
starch solution as indicator. If the oxygen is absent in the sample, the
MnSO4 react with the alkali to form white precipitate Mn(OH)2.

Relevant Indian Standard for Dissolved Oxygen Test :

1. IS 3025 (Part 38)-1989: Method of Sampling and Test (Physical

and Chemical) for Water and Wastewater, Dissolved Oxygen
(DO), First Revision.

PROCEDURE:
1. Fill a burette with a sodium thiosulphate solution (na2s2o3)
2. Take 10 mL of potassium dichromate ( k2cr2o7) in a conical
flask
3. Add 2 mL of hydrochloric acid (Hcl) and 1 spatula of potassium
iodide (KI) powder in the same conical flask
4. Titrate the sample with na2s2o3. Add 2 ml of starch when the
colour of the solution changes pale yellow. Then continue to
titrate till blue colour becomes colourless.
5. Now take another bottle and fill BOD bottle with sample water up
to the bottle brim
6. Immediately add 2 ml of manganese sulphate(MnSo4) solution
to the BOD bottle by inserting automatic pipette just below the
surface of the liquid.
7. Add 2 ml of alkali iodide azide reagent to the BOD bottle by
inserting automatic pipette just below the surface of the liquid
8. Mix the sample by inverting the BOD bottle several time allowing
precipitate to settle down at the bottom of the bottle
9. Add 2ml of concentrated sulphuric acid (H2So4) with a
graduated pipette carefully without forming any air bubble.
10. Take 200 ml of sample in a conical flask using a pipette.
11. Titrate the sample with na2s2o3. Add 2 ml of starch when
the colour of the solution changes pale yellow. Then continue to
titrate till blue colour becomes colourless
Conclusion / inference:
Acceptable limit of dissolved oxygen in water is 4mg/l to 7mg/l .
water with D.O less that 4mg/l is extremely harmful to drink. The
given water sample has D.0 =6.84 mg/l so it is in the BIS
standards range of drinking water.

EXPERIMENT-14
Aim: To determine the turbidity of the given sample using Nephelometer.

Principle:
The method presented below is based on a comparison of the intensity of light
scattered by the sample in specific conditions with the intensity of light
scattered by standard reference suspension under the same condition. The
higher the intensity of scattered lights, higher the turbidity. Formazine
polymer, which has gained acceptance as the turbidity standard reference
suspension is used as a reference turbidity standard suspension for water. It is
easy to prepare and is more reproducible in its lights scattering properties than
the clay or turbid natural water standards previously used. The turbidity of a
given concentration of formazine has an approximate turbidity of 100 NTU,
when measured on candle turbidity meter. Nephelometric turbidity units
based on formazine preparation will have approximate units derived from
Jackson candle turbidimeter but will not be identical to them.

Apparatus
Nephelometer with accessories

Reagents:
(i)Turbidity free distilled water (for setting zero).
(ii) Formazine turbidity concentrate (hydrazine sulphate + hexamine).
(iii) Formazine standard (for setting 100 of the instrument).

Preparation of Turbidity Free Distilled Water

Pass distilled water through a membrane filter having a precision pore size of
less than 10 microns (Whatman filter ). Rinse collecting flask at least twice with
such filtered water and discard the next 200 mL. Use this filtered water for
setting zero of the instrument.

Preparation of Formazine Turbidity Concentrate

(a) Solution I
Weigh accurately 5 g of ‘Anal–R’ quality hydrazine sulphate (NH2)2H2SO4 into
a 500 mL volumetric flask and add distilled water to make up to the mark.
Leave the mixture to stand for 4 hours.

(b) Solution II
Weigh accurately 50g of 'Anal-R' quality hexamethylene tetramine (CH2)6N4
(hexamine) into a 500 mL volumetric flask and add distilled water to make up
to the mark. Mix equal volume of solution I and II to form formazine turbidity
concentrate. Allow it to stand in a closed container at 25C to 30C for 48 hours
to produce insoluble white turbidity corresponding to 4000 NTU.
Note: Once prepared, formazine turbidity concentrate (which corresponds to
10000 ppm SiO2) is stable for 2 to 3 months.

Preparation of Formazine Standard

Dilute 25mL of the formazine turbidity concentrate to 1 litre with turbidity free
distilled water to obtain 250 ppm or 100 NTU for setting '100' of the
instrument.
Note: Formazine standard 100 NTU should be prepared weekly.

Procedure
(1) Switch the instrument on.
(2) Open the lid of the sample compartment.
(3) Insert a test tube filled with distilled water into the sample compartment.
Close the lid.
(4) Adjust 'SET 0' control to get '0' displayed on the read out.
(5) Open the lid. Replace the test tube filled with distilled water with a test
tube filled with formazine standard. Close the lid.
(6) Adjust the 'SET 100' control to get '100' displayed on the read out.
(7) Repeat the above operation to get consistent values of 0 to 100 within 1%
to 2%.

Measurement of turbidity less than 100 NTU

1. Thoroughly shake the sample.
2. Wait until air bubbles disappear and pour the sample into the nephelometer
tube.
3. Read the turbidity directly from the instrument.

Measurement of turbidity above 100 NTU

Dilute the sample with one or more volume of turbidity free distilled water
until the turbidity fall below 100 NTU.

A(B + C)
NTU of sample =
C
where,
A = NTU found in diluted sample
B = volume of dilution water in mL
C = sample volume taken for dilution in mL

-END-