Department of

Chemistry

National INSTITUTE of Technology

KURUkshetra

Energy and environmental Science

Lab maNUal
SUbject Code: CHLR11

Page 1 of 32
 InSTRUction to the STUdentS
Students must….

 Carefully read through the experiment to be performed.

 Look up information on equipment, materials and special techniques
required for the experiment.
 Bring your lab coat, Practical note book and observation note for all
the practrical class

Student instruction

 Wear chemical safety goggles and a knee length laboratory white coat
at all times while in the laboratory when anyone is conducting
experiments.
 Wear closed shoes at all times while in the laboratory.
 Wear nitrile gloves when directed to do so by your instructor and/or
lab manual.
 Confine long hair when in the laboratory so that it will not catch on
fire or come into contact with chemicals.

Behavioral Rules for Safety

 Do not eat, drink, smoke in the laboratory

at any time. Keep all food and drinks sealed
and in your backpack or purse.
 Consider all chemicals to be hazardous
unless instructed otherwise.
 Do not taste anything in the chemistry
laboratory.
 Do not use flammable liquids near open
flames. Most organic liquids are flammable.
 When heating substances in a test tube, never point the mouth of the
test tube at yourself or at anyone else. It may erupt like a geyser.
 Do not force glass tubing or thermometers into rubber stoppers. The
tubing or thermometer may break and cut you badly. Consult with
your laboratory instructor for assistance.

Page 2 of 32
 Use caution when handling Bunsen burners, hot plates, and glassware
or other equipment that has been heated. Burns are the most common
laboratory injury so treat all equipment as if it were hot during
experiments that involve heating.
 Do not perform unauthorized experiments.
 If you see someone else doing something you think may be
dangerous, tell him or her to stop and/or report the incident to your
lab instructor. If another student tells you to stop doing something
because it is unsafe, stop as directed. Consult your lab instructor if
there is a problem or difference of opinion.

Page 3 of 32
 LiST of ExperimenTS

1. Determination of alkalinity in the water sample

2. Determination of total Ca2+ and Mg2+ hardness in water.
3. Determination of dissolved oxygen (DO) in the water sample.
4. Determination of strength of nitrite ions in water samples.
5. Determination of residual chlorine in the water sample.
6. Determination of chemical oxygen demand (COD) in the water
sample.
7. Determination of total dissolved solids in water/effluent sample.
8. Analysis of BTX through Gas-Chromatography in air samples.
9. Determination of moisture and pH of soil sample
10. Analysis of heavy metal ions in industrial effluent by Atomic
Absorption Spectroscopy (AAS).
.

Page 4 of 32
 Experiment No. 1
Aim:
To determine the type and extent of alkalinity in the given water sample provided N/50
H2SO4.

Chemicals required:
N/50 H2SO4, water sample and Indicators

Apparatus required:
Conical flask, pipette and burette.

Indicators:

Phenolphthalein Methyl red

End point:
Based on the type of indicator

 Phenolphthalein (Pink to colorless)

 Methyl red (Yellow to red)

Theory:
Alkalinity of water is defined as its capacity to neutralize acid or alternatively the
amount of acid (gm/L) required in neutralizing one litre of alkaline water sample. The
alkalinity of such water sample is mainly due to hydroxide (OH¯), carbonate (CO3¯) or
bicarbonate (HCO3¯) ions. They are present alone or in combination with others. The
possible combinations are a) OH¯ only b) CO 32¯ only c). HCO3¯ only d) OH¯ with
CO32¯ and e). CO32¯ with HCO3¯. The combination of OH¯ with HCO3¯ is not possible
because they react to form water and carbonate ion.
OH + HCO3¯ H2O + CO32¯
This alkalinity of the water sample can be estimated by titrating a known amount of
water sample against a standard acid using phenolphthalein and methyl red as an
indicator. Phenolphthalein at basic condition shows pink color due to deprotonation of
two phenolic hrydrogens as shown scheme 1

Page 5 of 32
 Scheme 1
Therefore, the volume of acid used up to phenolphthalein end point A mL lead to
complete neutralization of OH¯ ion or half neutralization of CO32¯ ions. At the end
point color changes from pink to colorless at pH 8-8.5. The alkalinity measured up to
this point is known as phenolphthalein alkalinity. However Methyl red indicator at
acidic pH shows color change from yellow to red due to protonation as shown in
Scheme 2.

Scheme 2
The volume of acid used in further titration using methyl red indicator B mL,
corresponds to neutralization of HCO3¯ formed from half neutralization of CO 32¯
and/or present originally At this end point color changes from yellow to red at pH 4-5.
The total volume of acid used is A+B mL Therefore, this volume of acid corresponds to
complete neutralization of OH¯, CO32¯ and HCO3¯. It counts to total alkalinity or also
known as methyl red alkalinity. The type and extent of alkalinity may be determined by
comparing the value of A and B. The complete titration sequence is shown below

OH¯ + H+ H2O Phenolphthalein endpoint (A)

CO32¯ + H+ HCO3¯
HCO3¯ + H+ H2O + CO2 Methyl red endpoint (B)

Procedure:
Pipette out 20 ml of water sample and transfer into 250ml conical flask. Add 2-3 drops of
phenolphthalein indicator so that of water sample solution becomes pink. Titrate the
resulting pink color solution against N/50 acid (H2SO4) taken in the burette until color
changes from pink to colorless. Note the volume of acid (say A ml) consumed to reach
the phenolphthalein end point. Now, add few drops of methyl red indicator to the same
water sample so that the color of the solution becomes yellow. Now, continue the
titration of the resulting yellow solution against same N/50 acid until color changes from

Page 6 of 32
yellow to red. Note the volume of acid (say B mL) consumed to reach the methyl red
end point. Repeat the same process to get the concordant readings.

Observation table:

Titration of given water sample Vs N/50 H2SO4 solution.

S.No Initial Burette reading Volume of acid used (mL)

Burette Phenolphthalein Methyl red Phenolphthalein Methyl red end
reading end point (Y) end point (Z) end point point
(X ) A = Y-X B = Z-Y

1
2
3

Concordant reading: A = …….. , B = ......................

Calculation:

Volume of water sample used for each = 20 mL

titration
Volume of acid used for the total alkalinity = A+B mL
By applying normality equation, N1V1 (water sample) = N2V2 (acid solution)
Therefore, 20 x N1 = 1/50 x A+B or
N1= A+B/1000
Now, alkalinity of water sample (in terms = N1 x equivalent weight of CaCO3 in
of CaCO3 equivalents) mg/L.
Similarly, the alkalinity due to each ion separately can be calculated by following
relation between A and B.

Relationship OH CO32- HCO 3-

A=0 - - B

B=0 A - -

A=B - 2A=2B -

A>B A-B 2B -

A<B - 2A B-A

Page 7 of 32
Precautions:

 The burette should be rinsed with dilute H2SO4 acid.

 As H2SO4 acid is colorless solution, so lower meniscus should be taken as the correct
reading.
 Parallel error should be avoided while taken the readings.

Result:

1. Phenolphthalein alkalinity = .................. ppm

2. Methyl red alkalinity
3. Carbonate alkalinity
4. Bicarbonate alkalinity
5. Hydroxide alkalinity
6. Total alkalinity
= .................. ppm
= .................. ppm
= .................. ppm
= .................. ppm
= .................. ppm

Page 8 of 32
 Experiment No. 2

Aim
To determine the total hardness (Calcium and Magnesium) of water sample by using
complexometric titration method using standard EDTA solution.

Chemical requirement
 Hard water Sample (HWS)
 Ammonical buffer solution (NH4Cl + NH4OH)
 Calcium precipitating buffer [(NH4OH+NH4Cl+(COONH4)2]
 EDTA

Apparatus requirement
 Conical flask
 100 mL measuring flask
 funnel
 measuring cylinder
 500 mL beaker etc.

Indicator used:
Eriochrome Black T (EBT)

End Point:
Color changes from wine red to blue.

Theory:
The property of water which restricts or checks the lather formation with soap is called
hardness. In other words, the presence of multivalent cations, mostly calcium and
magnesium ions, in water is referred to as hardness of water. The Ca 2+ and Mg2+ ions
reacts with soap (Sodium or potassium salts of higher fatty acids) to form precipitate.

Mg/Ca (HCO3)2 + 2C17H35COONa (C17H35COO)2Ca/Mg + 2 NaHCO3

Sodium stearate Sticky White ppt.

Lather is produced only after all the hardness causing ions are completely precipitated in
the form of corresponding insoluble salts. Cleaning with soap in hard water is not
preferred as it causes a lot of wastage of soap. Hardness is generally expressed in terms of
CaCO3 equivalent and results are reported in ppm (Part per million). Where, l ppm =
1mg/L. Temporary or carbonate hardness which can be removed by boiling and
Permanent or noncarbonate hardness which cannot be removed by boiling. On the basis
of degree of hardness in water, it is classified as:
 Soft water = 0-75 ppm CaCO3 equivalent
 Moderately hard water = 75-100 ppm CaCO3 equivalent
 Hard water = 150-300 ppm CaCO3 equivalent
 Very hard water = >300 ppm CaCO3 equivalent

Page 9 of 32
The hardness of water can be determined by complexometric titration using Ethylene
diammine tetra acetic acid (EDTA). EDTA in the form of its di-sodium salt forms
complex with Ca2+ and Mg2+ ions of water sample.

EDTA (H 2Y2-) MY2- (M = Ca2+ or Mg2+)

For the simplicity the dibasic EDTA can be represented by H 2Y2- ions. It forms
complexes with Ca2+ and Mg2+ ions and other divalent cations (M2+) as represented
below:

H2Y2- + M2+ MY2- + 2H+

The complexes with hardness causing divalent ions are stable in alkaline medium (pH
11.5

11.0
range 8-10). The indicator used is Eriochrome Black T (EBT). It has two ionizable
phenolic hydrogen atoms. For simplicity it is represented by Na+ H2In¯

In the pH range of 7-11, the indicator reacts with metal ion to form a weak complex of
wine red color.

(blue) (wine red)

Buffer solution:

The optimum pH for the experiments is 10.0 and it is adjusted by using buffer
(NH4OH+NH4Cl) solution. A small amount of indicator solution is added to hard water
sample by controlling the pH with buffer solution. The indicator reacts with Mg2+ to
produce Mg-indicator complex which is wine red in color. Once EDTA is added, first
free calcium ions form most stable complex of the type CaY 2- (Ca-EDTA complex),
following by formation of Mg-EDTA complex which is less stable than Ca-EDTA
complex

Ca2+ + H2Y2- CaY2- + 2H+

Mg2+ + H2Y2- MgY2- + 2H+

Therefore, an extra drop of EDTA added after all the free Ca 2+ and Mg2+ ions have
formed the complex with the EDTA. Then, EDTA takes Mg 2+ from weak Mg- Indicator
complex to form stable Mg-EDTA complex and simultaneously releases the EBT

Page 10 of 32
indicator (HIn2-) in free form, Formation of this complex makes the end point of this
titration from wine red to blue.

MgIn- + H2Y2- HIn2- + MgY2- + H+

(wine red) (blue)
Determination of Mg and Ca Hardness : Calcium ions in hard water can be
2+ 2+

separated by converting them into white ppt. of the calcium oxalate by addition of Ca2+
precipitating buffer [NH4OH+NH4Cl+(COONH4)2] to give hard water having only
magnesium ions, as shown below.

Therefore, by precipitating and filtering off of Ca 2+ ions from hard water, Mg2+ ion
hardness can be calculated and from which Ca 2+ ion hardness can be determined by
subtracting the Mg2+ ion hardness from total hardness.

Procedure:

Step 1: Determination of total hardness:

 Pipette out 50 mL water sample in a titration flask and add 5 mL (small test tube)
of ammonical buffer solution along with 4-5 drops of EBT indicator.
 Titrate the resulting wine red color solution against standard EDTA solution till
the color changes to blue.
 Note the volume of EDTA solution consumed and repeat the experiment to get
concordant reading.

Step 2: Determination of Mg2+ ions hardness:

 Take 200 mL of hard water sample in a 500 mL beaker.
 Add 50 mL of Ca2+ precipitating buffer solution by measuring cylinder with
constant stirring. Keep the solution approx. for 30 min. to settle down the white
color precipitate.
 Filter the solution with the help of filter paper.
 Take 50 mL of filtrate in 250 mL conical flask and add 5 mL of ammonical buffer
solution along with 4-5 drops of indicator.
 Titrate the resulting solution against standard EDTA solution and note the
volume of EDTA consumed.
 Repeat the experiment to get concordant reading.

Step 3: Determination of Ca2+ Hardness:

 Ca2+ Hardness = Total Hardness - Mg2+ ions Hardness.

Page 11 of 32
Observations and calculations

Table 1: Determination of total hardness

S. No. Initial Burette Reading Final Burette Reading Volume of EDTA (mL)
1
2.
3.

Table 2: Determination of Mg2+ hardness

S. No. Initial Burette Reading Final Burette Reading Volume of EDTA (mL)
1
2.
3.

Calculation for Total Hardness

Concordant Reading = say A Therefore,
50 mL HWS = A mL of EDTA
1 mL HWS = A/50 mg of CaCO3 equivalents. (Because, 1 mL of EDTA = 1 mg of
CaCO3)
1L (or 1000 mL) HWS = A /50 x 1000mg of CaCO 3 equivalents = Total Hardness

Calculation for Mg2+ Hardness

Concordant Reading = say B
Total water sample = 250 mL (200 mL HWS + 50 mL Ca 2+ precipitating buffer)
Therefore, 250 mL of Water Sample = 200 mL HWS (OR)
1 mL of water Sample = 200/250 ml HWS
And, 50 mL Water Sample = 200/250 x 50 mL HWS = 40 mL HWS
So, 40 mL of HWS= B mL of EDTA = B mL of CaCO 3
1 mL HWS = B/40 mg of CaCO3 equivalents
1 L (or 1000 mL) HWS = B/40 x 1000 mg of CaCO 3 equivalents = Mg2+ Hardness.

Result

Total harness = ............... ppm

Mg ion hardness = ............. ppm
2+

Ca2+ ion hardness = ..............ppm

Precautions:

1. EDTA solution is colorless, so lower measuring should be taken as the

correct reading.
2. The burette should be rinsed properly.
3. Parallel error must be avoided while recording the reading.

Page 12 of 32
 Experiment No. 3
Aim
To estimate dissolved oxygen (DO) content in the given water sample by Winkler’s method
provided N/80 hypo solution

Chemical requirement
Conc. H2SO4, KMnO4, MnSO4 (40%), alkaline KI solution, N/50 hypo-solution and water
sample.

Apparatus requirement
Burette, Conical flask, measuring cylinder, BOD, bottle with stopper.

Indicator
Freshly prepared starch solution.

Structure of the starch (amylose and amylopectin) molecule

End point
Disappearance of blue colour.

Theory
Definition and environmental Significance: 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. In a healthy body
of water such as a lake, river, or stream, the dissolved oxygen is about 8 ppm. The minimum
DO level of 4 to 5 mg/L or ppm is desirable for survival of aquatic life.
 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 ppm.
 Oxygen is poorly soluble in water. Its solubility is about 14.6 for pure water at 0°C
under normal atmospheric pressure and it drops to 7 mg/l at 35°C.
 Higher temperature, biological impurities, Ammonia, Nitrates, ferrous iron, chemicals
such as hydrogen sulphide and organic matter reduce DO values.

Page 13 of 32
  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.

Principle
Dissolved Oxygen can be measured either by titrimetric or electrometric 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.
Winkler’s method: DO is determined by the titrimetric method developed by Winkler in
1888. This method is based on the fact that in alkaline medium D.O. oxidizes Mn2+ to Mn4+
which in turn oxidizes I - to free I2 in acidic medium, The released free I2 is then titrated with
standard N/50 hypo-solution using freshly prepared starch solution as a indicator added near
the end point. The consumed volume of hypo is equivalent to the D.O. present in the given
water solution.
Modified Winkler’s method: Sometimes the strength of D.O. altered in water due to the
impurities such as NO2-, SO32-, Fe2+. These ions as impurities also liberate I2 from KI. In the
modified Winkler’s method interference caused by oxidizing agent impurity (NO 2-) and
reducing agent like Fe2+ is removed by the addition of excess of KMnO4. This excess KMnO4
is than removed by the addition of potassium oxalate solution. The following reaction takes
place to remove these impurities:

NO2- + H2O NO3- + 2H+ + 2e- } × 5

MnO4- + 8H+ + 5e- Mn2+ + 4H2O } × 2
5 NO2 + 2MnO4- + 6H+ 5NO3- + 2Mn2+ + 3H2O

SO32- + H2O SO42- + 2H+ + 2e-} × 5

MnO4- + 8H+ + 5e- Mn2+ + 4H2O} × 2
5SO32- + 2MnO4 - + 6H+ 5 SO42- + 2Mn2+ + 3H2O

]x5
Fe2+ Fe3+ + e-
MnO4- + 8H+ + 5e- Mn2+ + 4H2O
5 Fe2+ + MnO4 - + 8H+ 5Fe3+ + Mn2+ + 4H2O

NO3- and SO42- formed above do not interfere, but Fe3+ ions interfere when present in amount
more than 10 mg/L and this interference can be stoped by converting Fe 3+ ions to poorly
dissociated complex FeF63- by adding KF.

Fe3+ + 6F- FeF63-

The excess of KMnO4 is destroyed by adding potassium oxalate (K2C2O4)

MnO4- + 8H+ + 5e- Mn2+ + 4H2O ]x2

5 C2O42- 10CO2 + 10 e-
- + 2-
2 MnO4 + 16H + 5 C2O4 2Mn2+ + 8H2O + 10 CO2

Page 14 of 32
 Then MnSO4 and alkaline KI are added to remove the Mn2+ ions as white ppt. of Mn(OH)2.
Mn2+ + 2 OH- Mn(OH)2 white ppt.

Oxygen dissolved in sample oxidizes Mn2+ to Mn4+ which is precipitated as brown hydrated
MnO.(OH)2.
Mn2+ + 4OH- MnO(OH)2 + H2O + 2e- } × 2
-
O2 + 2H2O + 4e 4OH-
2+ -
2Mn + 4OH + O2 MnO(OH)2

This is called fixation of oxygen. On acidifying the solution manganese (Mn4+) in higher
oxidation state oxidizes I- to free I2.
MnO(OH)2 + 4H+ + 2e- Mn2+ + 3 H2O
- -
2I I2 + 2e
+ -
MnO(OH)2 + 4 H + 2I Mn2+ + 3 H2O + I2

I2 released in the above step is titrated against standard hypo-solution using freshly prepared
starch solution as indicator.
2S2O32- S4O62- + 2e-
I2 + 2e- 2I-
2S2O32- + I2 S4O62- + 2I-
Procedure:

 Collect sample in a 300 mL BOD bottle

 Add 0.7 mL conc. H2SO4 followed by 0.2mL (4 drops) of KMnO4 solution to get the
pink colour solution
 Close the bottle by stopper and mix the content thoroughly by inverting it many times
and allow the solution to stand for few minutes.
 If the pink color of the solution disappears, then add 1-2 drops of KMnO4 solution
again. If the pink color persists than add 2-3 drops (0.5mL) of potassium oxalate
(K2C2O4) solution. Mix the content again and allow to stand for few minutes
 The pink color of the solution must be removed completely. A few drops of extra
K2C2O4 solution may be added if required.
 Now add 2 mL of MnSO4 solution followed by 3 mL of alkaline KI solution. Mix the
content once again and allow to stand for 20 to 25 minutes to settle the brown ppt of
MnO(OH)2.
 Add 1 mL of conc. H2SO4 to dissolve the precipitate completely.
 The color of the resulting clear solution become yellow
Take 100ml of this prepared solution in a 250 mL conical flask and titrate with N/50
hypo-solution until the color of the solution turned to pale yellow. At this stage add
around 2 mL of freshly prepare starch solution in one lot so that color of the solution
turned to blue. Titrate the resulting blue solution against the same hypo- solutions until
blue color disappeared. Note the volume of hypo solution consumed and repeat the
titration to get the concordant reading.

Page 15 of 32
Observation Table:
Titration of liberate Iodine vs N/80 hypo solution.

S. No. Initial Burette reading Final Burette reading Volume of hypo consumed (mL)
1
2
3

Concordant reading: A
Calculations:

N1V1 (prepared solution) = N2V2 (hypo solution)

N1 ×100 = 1/80 × A mL
N1 = A/8000
Strength of dissolved oxygen = (N1 x equivalent weight of oxygen x 1000) mg/L

Result:
Strength of Dissolved Oxygen = .......... mg/L or ppm.

Precautions:

1. Do not allow air to trap while sampling water during BOD analysis
2. The reagent should be added carefully and the bottle should be inverted 3-4 times to
mix the content. Well, it may be done in the sink.
3. Hypo-solution is a colorless solution, so lower meniscus should be taken as the
correct reading.
4. The freshly prepared starch solution should be added in one lot near the end point.
5. The water sample should not be exposed to air for a long time.
6. Parallax error must be avoided while taking the reading.

Page 16 of 32
 Experiment No. 4

Aim
To determine the strength of sodium nitrite (NaNO 2) in the given water sample provided
N/10 KMnO4 solution and N/10 Mohr’s salt solution.

Chemical required
N/10 KMnO4 solution, N/10 Mohr’s salt solution and 0.75 H2SO4

Apparatus required
Burette, pipette, funnel, 250mL flask, burette holder, measuring Cylinder

Indicator
KMnO4 acts as a self-indicator.

End-point
Disappearance of pink colour

Theory
Nitrites can be naturally or artificially occurring in groundwater. Nitrites come from
fertilizers through run-off water, sewage and mineral deposits. High levels of nitrites are
toxic to humans and animals, especially infants. It can enter the body as nitrate, a
nutrient which is essential to plant growth and be converted into nitrite, which disrupts
the oxygen delivering ability of hemoglobin in the bloodstream. Infants can develop a
life-threatening blood disorder known as blue baby syndrome if exposed to the nitrite
contaminated water. Therefore it is important to determine the strength of nitrite ion in
the water sample.
The strength of NaNO2 in a given water sample can be measured by titrating the given
nitrite solution against standard KMnO 4 solution in acidic medium. Acidified KMnO4
oxidizes nitrite ion to nitrate ion and it reduces to manganese ions as per the following
reactions:

5 NO2¯ + 5 H2O 5NO3¯ + 10 H+ + 10 e¯

2 MnO4¯ + 16H+ + 10 e- 2Mn2+ + 8 H2O

2MnO4¯ + 5NO2- + 6H+ 2Mn2+ + 5NO3 - + 3H2O

The following problems were observed

1. Nitrite ion is unstable in acidic medium and readily gets converted into nitrous
acid which is volatile.

NO2¯ + H+ HNO2

2HNO2 NO + NO2 + H2O

Page 17 of 32
 2. Environmental oxidation of NO2¯ may take place and produce an error in the
result.
3. The reaction between KMmO4 and NO2¯ is not instantaneous; hence accurate
reading cannot be obtained.

In order to avoid these problems, we may change our approach from direct to indirect
titration in which a known excess volume of KMnO 4 is taken in a conical flask in acidic
medium and temperature is raised up to approximately 400 C following by addition of a
known volume of nitrite solution. At this temperature, the reaction between KMnO 4 and
NO2- is fast and instantaneous as soon as we add the NO 2¯ in the conical flask.
The volume of KMnO4 is taken in slight excess and un-reacted KMnO4 is determined by
titrating the reaction mixture against N/10 Mohr’s salt solution. Acidified KMnO 4
solution being oxidizing agent oxidizes Fe2+ to Fe3+ and itself reduces to Mn2+ ions. The
redox reaction between Mohr’s salt and KMnO4 is as follows:

Fe2+ Fe+3 + e¯ ]x5

MnO4¯ + 8H+ + 5e- Mn2+ + 4H2O
5Fe2+ + MnO4 + 8H+ 5Fe +Mn2+ + 4 H2O
3+

In order to overcome the problem for the formation of volatile nitrous acid and
environmental oxidation of NaNO2 precaution should be taken that the tip of the pipette
should be dipped in the solution while slowly adding nitrite solution, so that no
atmospheric oxidation of NaNO2 takes place and NO2¯ ions do not get any time or any
chance to change to nitrous acid. In this way, we can determine the amount of KMnO 4
left unreacted and hence the amount of KMnO 4 used by NO2¯ ions.

Procedure
 Pipette out 10 mL of N/10 KMnO4 and transfer it to 250 mL conical flask
 Add 100 mL of 0.75 N H2SO4 with the help of measuring cylinder.
 Warm the resulting solution up to 400C (approx).
 Now slowly add 10 mL of nitrite solution to the solution in the conical flask by
keeping the tip of the pipette below the surface of the liquid.
 Titrate the resultant solution against N/10 Mohr’s salt solution till the disappearance
of pink colour.
 Note the volume of Mohr’s salt solution consumed
 Repeat the experiment to get concordant reading.

Observations

S. Initial burette Final burette Vol. of Mohr’s salt sol. Used

No. reading reading (mL)
1
2
3
Concordant reading = A
Page 18 of 32
Calculation:

Volume of N/10 KMnO4 solution taken each time = 10 mL

Volume of nitrite solution taken each time = 10 mL
Volume of Mohr’s salt solution consumed for the unreacted N/10 KMnO4 solution
reacting nitrite solution = (10 – A) mL
Therefore, Volume of N/10 KMnO4 = (10 – A) mL
Now, by applying normality equation
N1V1 (NaNO2) = N2V2 (KMnO4)
N1 x 10 = 1/10 x V2
N1 x 10 = 1/10 x (10 – A)
N1 = 10- A/100
Strength = N1 x Eq. Wt. of NaNO2 x 1000 mg/L

Result:
The strength of NaNO2 solution = ..........mg/L or ppm.

Page 19 of 32
 Experiment No. 5

Aim
To determine the total residual chlorine in a water sample

Chemical required
 10 % KI Solution
 Acetic Acid
 N/20 Sodium thiosulphate

Apparatus required
 Conical flask
 Burette
 Pipette

Indicator used
Freshly prepared starch solution

End Point
Disappearnce of blue colour

Theory
Environmental Significance and definition: Chlorination is primarily adopted to destroy
or deactivate disease-producing microorganisms in the public water supplies and
polluted rivers. Chlorine is usually added to water in gaseous form or as sodium or
calcium hypochlorite. When chlorine is added to water, some of the chlorine reacts first
with organic materials and metals in the water and is not available for disinfection (this
is called the chlorine demand of the water). The remaining chlorine concentration after
the chlorine demand is known as total chlorine.
Total chlorine is further divided into: 1) the amount of chlorine that has reacted with
nitrates and is unavailable for disinfection which is called combined chlorine and, 2) the
free chlorine, which is the chlorine available to inactivate disease causing organisms, and
thus a measure to determine the potability of water The word "residual" means
"remainder" or "that which is left", and as the name suggests the chlorine residual test is
used to measure the amount of chlorine remaining in the water at the time the test is
made. Excess Chlorination may produce adverse effects. Potentially carcinogenic chloro
organic compounds such as chloroform may be formed.
To fulfill the primary purpose of chlorination and to minimize any adverse effects, it is
essential that proper testing procedures be used. Several methods for measurement of
total residual chlorine are available including iodometric methods, amperometric
titration methods, and N,N-Diethyl-p-phenylenediamine (DPD) methods. We are going
to use Iodometric method for residual chlorine determination.

Page 20 of 32
Principle
The starch-iodide titration method, one of the oldest methods for determining chlorine,
is very non-specific for oxidants and generally is used for total chlorine testing at levels
above 1 mg/L Cl2. Chlorine will liberate free iodine from potassium iodide (KI)
solutions at pH 3-5(see equation). The liberated iodine is titrated with a standard solution
of sodium thiosulphate (Na2S2O3) with fresh starch as the indicator and adding neae the
end point . The end point of the titration is indicated by the disappearance of the blue
colored, starch-iodide complex.

Cl2 + 2KI 2KCl + I2

Na2S2O3 + I2 Na2S4O6 + 2NaI
I2 + Starch Blue Color

Procedure
 Pipette out 50 mL of water sample and transfer it to 250 mL conical flask
 Add 5 mL of 10 % KI and 1.5 mL of Acetic acid solution to maintain pH 3-5
 Cover the flask and mix the solution by shaking
 Titrate the resultant solution against N/20 hypo solution until the color of the
solution turned to pale yellow.
 At this stage add around 2 mL of freshly prepare starch solution in one lot so that
color of the solution turned to blue.
 At the end point blue color disappeared .
 Repeat the experiment to get concordant reading.

Observations

S. No. Initial burette reading Final burette reading Vol. of Hypo. Used (mL)
1
2
3

Concordant reading = A

Calculation

N1V1 (water) = N2V2 (hypo solution)

N1 × 50 = 1/20 × A mL
N1 = A/1000
Total chlorine residuals = (N1 x 35.5 x 1000) mg/L

Result

Amount of total residual chlorine in a given sample of water = ......... mg/L or ppm
Page 21 of 32
 Experiment No. 6

Aim
To determine the Chemical Oxygen Demand (COD) of a given water sample

Chemical required
 Potassium dichromate
 Sulfuric acid
 Ferrous ammonium sulphate (FAS)
 Silver sulphate
 Mercury sulphate
 Distilled water

Apparatus required
 Burette
 250 mL conical flask (Erlenmeyer Flask)
 Pipettes
 Wash Bottle

Indicator
Ferroin Indicator

End Point
Blue-green to wine red

Theory
Chemical oxygen demand (COD) is the measurement of the amount of oxygen in water
consumed for chemical oxidation of pollutants. The COD test is commonly used to
indirectly measure the amount of organic compounds in water. 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. COD
determines 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 the solution.
Environmental Significance and definition: COD is an important water quality parameter
because, similar to BOD, it provides an index to assess the effect of discharged waste on
the environment. Higher COD levels mean a greater amount of oxidizable organic
material in the water, which will reduce dissolved oxygen (DO) levels. A reduction in
DO can lead to anaerobic conditions, which is deleterious to higher aquatic life forms.
The COD test is often used as an alternate to BOD due to shorter length of testing time.

Page 22 of 32
Principle
The organic matter present in sample gets oxidized completely by potassium dichromate
(K2Cr2O7) in the presence of sulphuric acid (H2SO4), silver sulphate (Ag2SO4) and
mercury sulphate (HgSO4) to produce CO2 and H2O. The sample is refluxed with a
known amount of potassium dichromat in the sulphuric acid medium and the excess
potassium dichromate 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 O2 required to oxidize the organic matter. Chlorides are quantitatively
oxidized by dichromate and represent a positive interference. Therefore, Mercuric sulfate
is added to complex the chlorides, so that it does not interfere in the determination.

Procedure
The COD test procedure is based on the chemical decomposition of organic and
inorganic contaminants, dissolved or suspended in water. The result of a chemical
oxygen demand test indicates the amount of water-dissolved oxygen (expressed as parts
per million or milligrams per liter of water) consumed by the contaminants, during two
hours of decomposition from a solution of boiling potassium dichromate. The higher the
chemical oxygen demand, the higher the amount of pollution in the test sample.
 Place 50.0 mL of sample in a 500 mL refluxing flask.
 Add 1 g of mercuric sulphate and a few glass beads.
 Add 5 mL of sulphuric acid-Ag2SO4 to dissolve the mercuric sulphate and cool.
 Add 25.0 ml 0.25 N potassium dichromate solution and mix well
 Attach the flask to the condenser and start the cooling water.
 Add the remaining acid reagent (70 mL) through the open end of condenser and
reflux for 2 hours.
 Cool and wash down the condenser with distilled water.
 Dilute the mixture to about twice its volume and cool to room temperature.
 Titrate the excess dichromate with standard ferrous ammonium sulphate using
ferroin indicator (2 to 3 drops).
 The color change from blue green to wine red indicates the end point.
 Same procedure repeated for blank consisting of distilled water of equal volume
as that of the sample

Page 23 of 32
Observations

S. Initial burette Final burette Volume of FAS

No. reading reading (mL)
Sample 1
2
3

Concordant reading = A mL

S. Initial burette Final burette Volume of FAS

No. reading reading (mL)
Blank 1
2
3

Concordant reading = B mL

Calculation
Volume of Sample = 50 mL
Volume of FAS used in the sample titration = A mL
Volume of FAS used in the blank titration = B mL
Therefore,
Volume of 0.25 N FAS equivalent to K 2Cr2O7 used for COD determination = B-A mL

N1V1 (water) = N2V2 (FAS solution)

N1 x 50 mL = 0.25 × (B-A) mL
N1 = 0.25 × (B-A) mL /50
COD = 0.25 × (B-A) mL /50 × 8 g/L
= 0.25 × (B-A) mL /50 × 8 × 1000 mg/L
= (B-A) × 40 mg/L

Result
Amount of DOD in a given sample of water = ......... mg/L or ppm

Precaution

 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

Page 24 of 32
 Experiment No. 7

Aim

Determination of total dissolved solids in water/effluent sample.

Apparatus required

 Evaporating Dish
 Water Bath
 Oven
 Desiccators
 Analytical Balance
 Gooch Crucibles
 Filter
 Pumps
 Forceps

Theory:
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. 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-
181 C. The increase in dish weight represents the total dissolved solids. Water can be
classified by the amount of TDS per litre: fresh water < 1500 mg/L , brackish water 1500
to 5000 mg/L and saline water> 5000 mg/L.

Environmental Significance:

 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.
Page 25 of 32
  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.

Procedure:

 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. Let’s denote the
weight measured as W1 = X g.
 Mix sample well and pour into a funnel with filter paper. Filter approximately 80
-100 mL of sample.
 Using pipette transfer 50 mL of unfiltered sample in the porcelain dish.
 Switch on the oven and allowed to reach 105°C. 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
.

Page 26 of 32
Observation Table

Description Weight
Weight of the clean porcelain evaporating dish (W1) X g
Weight of the dish and the residue (W2) Yg

Weight of residue (W) Y-X g

Volume of the Sample (V) 50 mL

Calculation
Weight of the clean porcelain evaporating dish (g) W 1 =X g
Weight of the dish and the residue (g) W2 =Y g
Weight of residue (g) W = Y- X g
The volume of the sample (mL) V = 50.0 mL
Total Dissolved Solids (TDS) = (Y-X ) / 50 X 1000 mg/L

Result

Amount of total dissolved solids in water/effluent sample is = ......... mg/L or ppm

Page 27 of 32
 Experiment No. 8

Aim: BTX analysis through gas-chromatography (GC)

Page 28 of 32
Page 29 of 32
Page 30 of 32
 Experiment No. 9

Aim:
Determine the moisture content of the given soil sample.

Theory:
In almost all soil tests natural moisture content of the soil is to be determined. The
knowledge of the natural moisture content is essential in all studies of soil mechanics. To
sight a few, natural moisture content is used in determining the bearing capacity and
settlement. The natural moisture content will give an idea of the state of soil in the field.
The natural water content also called the natural moisture content is the ratio of the
weight of water to the weight of the solids in a given mass of soil. This ratio is usually
expressed as percentage.

Apparatus required

1. Non-corrodible air-tight container.

2. Electric oven, maintain the temperature between 1050 C to 1100 C.
3. Desiccator.
4. Balance of sufficient sensitivity.

Procedure

 Clean the container with lid dry it and weigh it (W1).

 Take a specimen of the sample in the container and weigh with lid (W2).
 Keep the container in the oven with lid removed. Dry the specimen to constant
weight maintaining the temperature between 1050 C to 1100 C for a period
varying with the type of soil but usually 16 to 24 hours.
 Record the final constant weight (W3) of the container with dried soil sample.
Peat and other organic soils are to be dried at lower temperature (say 600 )
possibly for a longer period.

Certain soils contain gypsum which on heating loses its water if crystallization. If it is
suspected that gypsum is present in the soil sample used for moisture content
determination it shall be dried at not more than 800 C and possibly for a longer time.

Page 31 of 32
Observation and Calculation

Result

The natural moisture content of the soil sample is

Precautions
 A container without lid can be used, when moist sample is weighed immediately
after placing the container and oven dried sample is weighed immediately after
cooling in desiccator.
 As dry soil absorbs moisture from wet soil, dried samples should be removed before
placing wet samples in the oven.

Page 32 of 32