ANALYTICAL PROCEDURES OF WATER

1. Turbidity (Ref- APHA 2130 B)

Summary:

Turbidity is a term used to indicate the extent of cloudiness or muddiness of water

and muddiness caused in one litre of water by 1 mg of kaolin is taken as a turbidity
of 1 ppm.

Turbidity procedure by Turbidity Meter:

In this method, the turbidity of water is measured by using Turbidity Meter. Sample
is taken in Turbidity cell after thoroughly shaking and placed in Turbidity meter.
Measure the displayed value.

Apparatus required:
1. Turbidity meter

Turbidity standard 4000 NTU :

The particles of formazin are uniform in size and shape. The stock standards
prepared from formazin are accurate with in 1% and stable for 6-8 months

Preparation:

Solution A:

Take 5.0 grams of reagent grade “Hydrazine sulphate” and dissolve in 400ml of
distilled water.

Solution B:

Dissolve 50 grams of pure “HexamethyleneTetramine” in 400ml of distilled water.

 Mix solution A&B and make it up to one litre by adding distilled water and allow this
mixture to settle for 48 hours at normal room temperature.

This is stock solution of 4000 NTU strength of Formazin. This solution is usually
stable for a period of 6-8 months and working standard can be prepared as required
using following table.

Volume of stock solution (ml) to Final strength of Formazin in

make up 1 litre with higher grade diluted stock solution (NTU)
distilled water

125 500

25 100

12.5 50

20 of 500 NTU Solution 10

10 of 500 NTU Solution 5

Shake this stock solution well before dilution.

Cooling Water Parameters 2 Analytical Procedures

2. M-Alkalinity (Ref- APHA 2320 B)

Summary:

The alkalinity of water is either expressed by the equivalent of an acid in ppm that
is required in the neutralization of alkaline salts such as bicarbonate salt, carbonic
acid salt, hydroxyl compounds etc. dissolved in the water or expressed in ppm by
converting into the weight of calcium carbonate corresponding to the acid. The M-
alkalinity is measured by titrating the test water with N/50 sulphuric acid, the
mixed methyl red indicator being employed. (pH=3.8)

Reagents:

1. Mixed methyl red indicator :

Dissolve 0.02g of methyl red and 0.1g of bromocresol green in 100 ml of alcohol
(95%v/v).

2. N/10 Sodium carbonate solution (For standardization):

Heat in advance sodium carbonate (Na2CO3)-(a standard reagent for
O
quantitative analysis) at 100-110 C for 40-50 minutes; after leaving it in a
desiccator containing sulphuric acid, weigh out 5.300g of 100% Na 2CO3 and
dissolve it in water; place the mixture in a 1 litre volumetric flask and add water
up to the mark.

3. N/10 Sulphuric acid:

Add 3 ml of sulphuric acid to the beaker in which 100 ml of water has been
placed before hand and shake the solution thoroughly; after cooling it, add
distilled water to increase it to 1 litre.

4. N/50 Sulphuric acid:

Place 200 ml of N/10 sulphuric acid in 1 litre volumetric flask, add water that is
free of carbonic acid to get 1 litre. 1 ml of this solution is equivalent to 0.02
ppm corresponding to 1 mg converted in terms of calcium carbonate.

Cooling Water Parameters 3 Analytical Procedures

 5. Determine exact concentration by standardizing with std. Sodium carbonate
solution. Standardization of H2SO4: Take 25 ml of 0.02N Sodium Carbonate
solution in a conical flask add 3 drops of mixed indicator. Titrate against
N/50Sulfuric acid until the solution turns to reddish purple.
Normality of H2SO4 = 25 X 0.02/ Titre value of H2SO4.

Operating Method:

1. Place in a beaker 100 ml of water to be examined (when turbid, use the

supernatant liquid by filtering through the filter paper (Whatman No.42) or by
centrifugally separating it. Add 3-5 drops of mixed methyl red indicator.

2. If the water turns blue, it means that there is a presence of some hydroxide,
carbonic acid salt or carbonate; accordingly titrate it with N/50 sulphuric acid
until the solution turns reddish purple (pH 3.8), subsequently finding the M –
alkalinity.

Calculation :

M alkalinity in PPM as CaCO3 = Titre value X Normality of H2SO4X1000X50

Sample taken

Cooling Water Parameters 4 Analytical Procedures

3. P-Alkalinity water (Ref- APHA 2320 B)

Summary :

In this method, P-alkalinity (pH8.3) of water is measured when pH of water is more

than 8.3, by titrating with N/50 sulphuric acid using mixed phenolphthalein
indicator.

Reagents :

1. Phenolphthalein indicator :
Dissolve 0.5 gm of phenolphthalein in 50 ml of Iso PropylAlcohol and 50 ml
distilled water.

2. N/50 Sulphuric Acid:

Add 3 ml of sulphuric acid to the beaker in which 100 ml of water has been
placed before hand and shake the solution thoroughly; after cooling it, add
distilled water to increase it to 1 litre.

Operation:

1. Collect 100 ml of water to be tested in a beaker and add 4 drops of

phenolphthalein indicator.
2. When the water to be tested turns pink, it is an indication of the presence of
carbonate or hydroxide. Titrate the solution with N/50 sulphuric acid until the
pink color turns colorless (pH 8.3). Thus getting the P-alkalinity.

Calculation:

P alkalinity in PPM as CaCO3= Titre value X Normality of H2SO4X1000X50

Sample taken

Cooling Water Parameters 5 Analytical Procedures

4. Total Hardness (Ref- APHA 2340 A)

Summary:

Hardness of water is represented by the amount (ppm) of calcium carbonate

corresponding to the amount of calcium and magnesium ions in the water. After
adding a buffer to a sample to adjust it at about pH 10, it is titrated using
Eriochrome black T (E.B.T.) as indicator, with disodium ethylenediamine tetra acetic
acid (E.D.T.A) to get the total hardness.

Reagents:

1. Buffer solution:
Dissolve 1.179 gm of sodium salt of EDTA dihydrate and 644 mg of MgCl 2.6H2O in
50ml distilled water. Add this solution to 16.9 gm of ammonium chloride (NH 4Cl)
in 143 ml of liquid ammonia (NH4OH) and dilute it with distilled water to 1 litre.
Keep this solution in storage, tightly plugged up.

2. Eriochrome black T indicator:

Dissolve 0.5 gm Eriochrome black T in 100 ml of methyl alcohol. Keep it in
amber colored bottle.

3. Standard calcium chloride solution:

Weigh out 1.0 gm of CaCO3 or 1.470 gm of calcium chloride (CaCl 2. 2H2O) and
dissolve it in approximately 2 to 5 ml of hydrochloric acid; transfer it to a 1 litre
volumetric flask and dilute it up to the mark with distilled water.

4. Standard E.D.T.A. solution (0.01M) / 0.02N :

Dissolve about 3.723 gm of AR Grade disodiumethylenediaminetetra acetic acid
(Na2EDTA. 2H2O) in distilled water and dilute to one litre.
For standardizing this solution measure out 25 ml of standard calcium chloride
solution in a 250ml conical beaker, and dilute it with water to 50 ml and add 10
ml of buffer solution and 3 to 4 drops of E.B.T. indicator. Titrate with E.D.T.A.
solution until the reddish color changes to blue.

Cooling Water Parameters 6 Analytical Procedures

 Normality of EDTA = 25 X.02
X
Where ,
X = E.D.T.A. used in titrating (ml)

5. Inhibitor for Heavy metals:

Dissolve 5.0 gm sodium sulfide (Na2S.9H2O) in 100 ml distilled water.

6. Procedure:
a. Pipette out 25 ml of a sample into a 250 ml conical beaker.
b. Add 1 ml of ammonia buffer solution and 1 to 2 drops of E.B.T. indicator (pH
10).
c. While shaking and mixing well, titrate with standard E.D.T.A. solution; and the
point where the reddish color converts to blue color is taken as the end point.
Read the quantity in ml of E.D.T.A. consumed in titrating.

7. Calculation:

Total hardness (CaCO3) PPM = Titre value X Normality of EDTA X 1000 X 50

Sample taken

Note:

If heavy metals like Iron are high, add 1 ml of inhibitor solution in hardness test.
If alkalinity is more than 300 ppm, take 25ml water sample add 1ml HCl and boil
for half an hour. Cool the sample and proceed for hardness analysis.

Cooling Water Parameters 7 Analytical Procedures

5. Calcium Hardness (Ref- APHA 3500 Ca-D)

Summary:

In this method, after having made pH of water sufficiently high, it is titrated with
disodiumethylenediamine tetra-acetic acid (E.D.T.A.) liquid by using Calcon as
indicator.

Reagents:

a. Indicator:
Weigh 0.5 gm Calcon indicator mix in a pestle mortar with 5 gm of sodium
sulphate (anhydride) or 5 gm of Potassium chloride thoroughly until the mixture
becomes uniform or homogeneous.

b. Potassium hydroxide solution or Sodium hydroxide solution:

Dissolve about 225 gm of KOH in 500 ml of distilled water or 40 gm of NaOH in
500 ml of distilled water. Keep the solution in polyethylene bottle for
preservation.

c. Standard calcium chloride solution:

Weigh out 1.0 gm of CaCO3 or 1.470g of calcium chloride (CaCl 2. 2H2O) and
dissolve it in approximately 2 to 5 ml of hydrochloric acid; transfer it to a 1 litre
volumetric flask and dilute it up to the mark with distilled water.

d. Standard E.D.T.A. solution (0.02 N)

Dissolve about 3.723 gm of AR Grade disodium ethylenediamine tetra acetic acid

(Na2EDTA. 2H2O) in distilled water and dilute to one litre. For standardizing this
solution measure out 25 ml of standard calcium chloride solution in a 250ml
conical beaker, and dilute it with distilled water to 50 ml.Add10 ml of buffer
solution and 3 to 4 drops of E.B.T. indicator.

Titrate with E.D.T.A. solution until the reddish color turns to blue.

Cooling Water Parameters 8 Analytical Procedures

 Normality of EDTA = 25X.02
X
Where ,
X = E.D.T.A. used in titrating (ml)

e. Hydroxylamine chloride solution (10%)

f. Inhibitor for Heavy metals:

Dissolve 5.0 g sodium sulfide (Na2S.9H2O) in 100 ml distilled water.

Procedure:

1. Collect 25 ml of a sample in a 250 ml conical flask. (If turbid, filter it using

Whatman filter paper no.44). To this solution add 1.0 ml of potassium
hydroxide solution or 1.0 ml of sodium hydroxide solution; and add 1 spec
(pinch) of Calcon.

2. Titrate with standard E.D.T.A. solution until the reddish color turns to blue color.

Calculation :

Calcium Hardness (CaCO3) PPM = Titre value X Normality of EDTA X 1000 X 50

Sample taken

Note:

If heavy metals like Iron are high, add 1 ml of inhibitor solution in Calcium test.
If alkalinity is more than 300ppm, take 25ml water sample add 1ml HCl and boil for
half an hour. Cool the sample and proceed for calcium hardness analysis.

Cooling Water Parameters 9 Analytical Procedures

6. Magnesium hardness

Summary:

The magnesium hardness of water is calculated by subtracting its calcium hardness

from the total hardness of the water.

Calculation:

Magnesium hardness in (CaCO3) ppm = Total hardness in (CaCO3) ppm –

Calcium hardness in (CaCO3) ppm

Cooling Water Parameters 10 Analytical Procedures

7. Phosphate ions by Metol method (Total PO4 including organic phosphates)
(Ref- BIS 1109, modified)

Summary:

In this method, a heteropolymer compound obtained by adding ammonium

molybdate in acid medium is reduced by metol causing the solution to develop
molybdenum blue colour, the absorbance of which is measured to estimate the
phosphate ions in water.

Apparatus required:
1. Spectrophotometer

Reagents:

1. Metol:
Dissolve 20 gm metol in a 200 ml water _______________________ (1)
100 gm sodium metabisulphite dissolved in 200ml water __________ (2)
20 gm citric acid dissolved in a 200 ml water ____________________ (3)
Mix (1),(2) & (3) in 1 litre standard flask and dilute to one litre.

2. Ammonium-Molybdate Solution:
Dissolve 50 gm ammonium molybdate in 200ml distilled water _______ (A)
50 ml Conc. H2SO4 in 400 ml distilled water _____________________ (B)
Cool and mix (A) & (B) and dilute to one litre.

3. (1 + 2) H2SO4 :
Mix one part of sulphuric acid carefully and slowly in two parts of distilled water.

Operating procedure:

1. Take 25ml sample in 100 ml volumetric flask, add 1 ml (1+2) H2SO4 and 0.5
gms Ammonium persulphate and digest for 15-30 minutes. Add distilled water
and do not allow to dry. Cool the solution in water bath and add 5.0 ml of metol
solution. Add 5.0 ml of ammonium molybdate solution. Make up to 100 ml and
find absorbance/concentration after 30 minutes with 10mm cell at 720nm. Run
blank also.

Cooling Water Parameters 11 Analytical Procedures

 Calculation

Total Phosphates as PO4 = (Sample value - Blank value) X Total volume

Sample volume

Note-
For total inorganic (ortho & Pyro) PO4, add acid and digest but do not
add ammonium persulphate.

Poly phosphate as PO4 = Total inorganic phosphate – Ortho phosphate

Organic phosphate as PO4 = Total phosphate – Total inorganic phosphate.

Calibration :

1. Dissolve 1.4325 g of KH2PO4 in 1 litre of DM water; it is equivalent to 1000

ppm of PO4.

2. Dilute 10 ml solution (from A) to 100 ml with DM Water; it is equivalent to 100

ppm of PO4

3. Take 1,2,3,4,5,6,7,8,9 and 10 ml from solution B separately in to a 100 ml

volumetric flask.

a. Add 1 ml of H2SO4 + 5 ml METOL + 5ml Amm. molybdate and make up to

100 ml with DM Water

b. Wait for 20 to 30 minutes, record the absorbance on a Spectrophotometer

at 720 nm and plot the graph absorbance Vs Concentration.

Cooling Water Parameters 12 Analytical Procedures

8. O-Phosphate by metol method

Summary:

In this method, a heteropolymer compound obtained by adding ammonium

molybdate in acid medium is reduced by metol causing the solution to develop
molybdenum blue colour, the absorbance of which is measured to estimate the
phosphate ions in water.

Apparatus required:
1. Spectrophotometer

Reagents:

1. Metol:
Dissolve 20 gm metol in a 200 ml water _______________________ (1)
100 gm sodium metabisulphite dissolved in 200ml water __________ (2)
20 gm citric acid dissolved in a 200 ml water ____________________ (3)
Mix (1),(2) &(3) in 1 litre standard flask and dilute to one litre.

2. Ammonium-Molybdate Solution:
Dissolve 50 gm ammonium molybdate in 200ml distilled water _______ (A)
50 ml Conc. H2SO4 in 400 ml distilled water _____________________ (B)
Cool and mix (A) & (B) and dilute to one litre.

4. (1 + 2) H2SO4 :
Mix one part of sulphuricacid carefully and slowly in two parts of distilled water.

Operating procedure:

2. Take 25 ml sample and (blank) in 100 ml volumetric flask, add 1 ml (1+2)

H2SO4, 5 ml of metol solution and 5ml of ammonium molybdate solution. Make
up to 100 ml and find absorbance after 30 minutes with 10 mm cell at 720nm.

Cooling Water Parameters 13 Analytical Procedures

 Calculation

O- Phosphates as PO4 = (Sample value - Blank value) X Total volume

Sample volume

Calibration :

1. Dissolve 1.4325 gm of KH2PO4 in 1 litre of DM water; it is equivalent to 1000

ppm of PO4.

2. Dilute 10 ml solution (from A) to 100 ml with DM Water; it is equivalent to100

ppm of PO4

3. Take 1,2,3,4,5,6,7,8,9,and 10 ml from solution B separately in to a 100 ml

volumetric flask.

a. Add 1 ml of H2SO4 + 5 ml METOL + 5ml Ammonium molybdate and make

up to 100 ml with DM Water

b. Wait for 20 to 30 minutes, record the absorbance on a Spectrophotometer

at 720 nm and plot the graph. Graph will be absorbance Vs Concentration.

Cooling Water Parameters 14 Analytical Procedures

9. Determination of Zinc (Ref- APHA 3500 D-I, modified)

Zinc concentrations in most water supplies average about 1 mg / litre but may
range as high as 50 mg / litre in some areas. Although zinc commonly is found in
many natural waters, the deterioration of galvanised iron & leaching of brass can
add substantial amounts. Industrial effluents may contribute large amounts of zinc.
In cooling water treatment zinc salts are used as Cathodic Inhibitors along with
other inhibitors.

SUMMARY OF METHOD

Zinc reacts with dithiozone indicator in weakly medium to form a red compound.
The zinc – dithiozone reaction is extremely sensitive. Most interference can be
overcome be adjusting the pH to 4.0 to 5.5 & by adding sufficient sodium
thiosulfate. The zinc is estimated by complexometric titration using Disodiium salt
of EDTA and Dithiozone indicator in an Acetate Buffer medium.

REAGENTS

i. ACETATE BUFFER SOLUTION :

Dissolve 270 gram Sodium Acetate in distilled water. Add 225 ml of Glacial Acetic
Acid (Analar / Excelar Grade). Mix it well. & make up to 1000 ml with distilled
water.

ii. DITHIZONE INDICATOR SOLUTION :

Dissolve 0.05 gram of Dithiozone indicator powder in 100 ml Acetone. Keep the
solution in a stoppered to avoid loss of Acetone.

iii. EDTA SOLUTION :

Weigh accurately 0.575 gram of Ethylene diamine tetra acetic acid, disodium salt,
dihydrate Na2EDTA. 2H2O (Analar / Excelar Grade). Dissolve it in water. Add 5 ml
1N sodium Hydroxide Solution & make upto 1000 ml with Distilled water.
1 ml of this solution will correspond to 0.1 ppm zinc.

Cooling Water Parameters 15 Analytical Procedures

iv. ISOPROPYL ALCOHOL : Analar / Excelar Grade

v. SODIUM THIOSULFATE SOLUTION :

Dissolve 49.7 gram Sodium thiosulfate in Distilled water. Add 0.1 gram sodium
carbonate & make upto 1000 ml with Distilled water.

PROCEDURE

1. Pipette out 100 ml filtered water sample in a 250 ml conical flask.

2. Add 5 ml Acetate Buffer solution followed by 1 ml sodium thiosulfate solution. Mix
it well.
3. Add 50 ml Isopropyl Alcohol & 1 ml Dithizone indicator solution. Mix it well.
4. A pink color indicates the presence of zinc.
5. Titrate with EDTA solution until the color changes from pink to blue violet.
6. Note down the reading of EDTA consumed in the titration R ml.

Zinc R x 100
as = ml sample
ppm Zn

Where
R = ml of EDTA consumed in the titration.

Test Kit Procedure :

1. Take 5 ml sample in the easy Test Jar

2. Add 5 drops of Zinc - 1A Reagent and mix well.
3. Add 2 Scoops (0.5g) of Zinc -2A Reagent. Shake the Jar until it dissolves
completely.
4. Wait for 5 minutes.
5. Add 3 drops of Zinc-3A Reagent and mix. Color develops.
6. Compare color intensity immediately (within 5 minutes) with color chart.

Note:
For more details see Booklet which we provided along with Test Kit.

Cooling Water Parameters 16 Analytical Procedures

10. Total Iron (Ref- APHA 3500 Fe-D)

Summary:
Iron is present in water as bivalent ions: this compound has a comparatively large
solubility. Bivalent iron ions are oxidized into trivalent state after being exposed to
air becoming iron hydroxide having low solubility. In this method, total iron ions
present in the water are reduced wholly into ferrous ions by hydroxyl ammonium
chloride and then Phenanthroline solution is added to the above to form reddish
orange complex, the absorbance of which is subsequently measured for the
estimation of total iron content.

Apparatus required
1. Spectrophotometer

Reagents :

1. Hydroxyl Ammonium Chloride:

Dissolve 10 gm NH2OH.HCl in 100 ml distilled water.

2. Phenanthroline solution
Dissolve 100 mg of 1,10-Phenanthroline monohydrate C12H8N2.H2O in 100ml of
distilled water by stirring.

3. Ammonium acetate/acetic acid buffer (pH 4.2 approx.)

Dissolve 100 gm of ammonium acetate by adding 60 ml of distilled water and
280 ml of glacial acetic acid.

4. Stock Iron Solution

Add slowly 20 ml of concentrated H2SO4 to 50 ml distilled water. Dissolve 1.404
gm of Ferrous ammonium sulphate Fe(NH4)2 (SO4)2.6H2O and 0.1N KMnO4 drop
wise until a faint pink color persists. Dilute to 1000 ml with iron free distilled
water.
1 ml = 200 ppm Fe

5. Standard Iron Solution (Prepare Daily)

Dilute 50 ml stock solution to 1000 ml in a volumetric flask with iron free
distilled water. 1 ml = 10 ppm Fe

Cooling Water Parameters 17 Analytical Procedures

 Operating procedure

1. Place 50 ml of uniformly mixed water sample to be examined (containing 0.005

– 0.5 mg as Fe) in a conical flask; add 1 ml of hydrochloric acid and heat to boil
the solution for 30 minutes or until it is reduced to about less than 20 ml.

2. After cooling, add distilled water if the volume is less than 20ml. Then filter the
sample through Whatman filter paper in a 100 ml volumetric flask.

3. Add 5.0 ml of freshly prepared hydroxyl ammonium chloride and 4.0 ml of

Phenanthroline solution and mix them by shaking. Add 10 ml of ammonium
acetate-acetic acid buffer solution and make up the volume to 100 ml. Mix them
again by shaking, Allow the solution to stand for about 20 minutes.

4. Record absorbance by spectrophotometer at wave length 510nm.

Note
Throughout the whole operating procedure, conduct blank tests and correct the
result previously obtained.

Calculation

Total Iron as Fe = (Sample value-Blank value) X total volume/sample volume.

Calibration Procedure:

1. Take 1 ml ,2,3,4,5,6,7,8,9,10 ml standards from 10 ppm Fe standard in 100 ml

volumetric flasks separately. Add 50 ml distilled water.

2. Add 5.0 ml of freshly prepared hydroxyl ammonium chloride and 4.0 ml of

Phenanthroline solution and mix them by shaking. Add 10 ml of ammonium
acetate-acetic acid buffer solution and make up the volume to 100ml. Mix them
again by shaking. Allow the solution to stand for about 20 minutes.

3. Record absorbance on Spectrophotometer at wave length 510 nm.

Cooling Water Parameters 18 Analytical Procedures

11. Dissolved Iron (Ref- APHA 3500 Fe-D)

Reagents:
1. Hydroxyl Ammonium Chloride:
Dissolve 10 gm NH2OH.HCL in 100ml distilled water.
2. Ammonium acetate buffer solution :
Dissolve 100 gm NH4C2H3O2 (Ammonium Acetate) in 60ml distilled water. Add
280ml conc. (glacial) acetic acid.
3. Phenanthroline solution :
Dissolve 100mg 1,10-phenanthroline monohydrate, C12H8N2.H2O in 100ml
distilled water by stirring.
4. Stock Iron Solution:
Add slowly 20ml conc. H2SO4 to 50 ml distilled water and dissolve 1.404 g
Fe(NH4)2 (SO4)2.6H2O. Add 0.1 N KMnO4 (3.2 g/l) drop wise until a faint pink
color persists. Dilute to 1000 ml with iron free distilled water. 1ml = 200 ppm
Fe.
5. Standard Iron Solution:
(Prepare daily) Dilute 50ml stock solution to 1000ml in a volumetric flask with
iron free distilled water. 1ml = 10 ppm Fe.

Procedure:
Take 50 ml sample in a standard flask add 5 ml Hydroxyl ammonium chloride 10 ml
Acetate buffer 4 ml phenanthroline and make up to 100 ml. Wait for 20 minutes.
Record absorbance, at 510 nm using spectrophotometer.
Calculation
Dissolved Iron as Fe = (Sample value-Blank value )X total volume/sample volume.

Calibration Procedure:
1. Take 1 ml ,2,3,4,5,6,7,8,9,10 ml standards from 10 ppm Fe standard in 100 ml
volumetric flasks separately.
2. Add 5.0 ml of freshly prepared hydroxyl ammonium chloride and 4.0 ml of
Phenanthroline solution and mix them by shaking. Add 10ml of ammonium
acetate-acetic acid buffer solution and make up the volume to 100 ml with
distilled water. Mix them again by shaking. Allow the solution to stand for about
20 minutes.
3. Record absorbance by spectrophotometer at wave length 510 nm and plot a
Graph.

Cooling Water Parameters 19 Analytical Procedures

12. A) Chloride (Ref- APHA 4500 Cl—C)

Requirements:

1. 0.2N NaCl solution:

Dry NaCl AR grade in oven at approximately 105°C .Weigh and dissolve
1.17gm of dried NaCl into DM water in a volumetric flask and make the
final volume to 100 ml.
2. 0.02N NaCl solution:
Pipette out 10 ml of 0.2N NaCl solution to 100ml with DM water.
3. 0.02N Hg (NO3)2 solution :
7.46 0.02 gm of Mercuric Nitrate AR grade are dissolved in DM water
along with 6 ml of conc. Nitric acid LR. The final solution is made up to
2000 10 ml shake well and standardize against 0.02N NaCl solution.
4. 0.2% w/v Diphenyl Carbozone indicator (DPC):
Dissolve 2 0.02g of DPC in 1 litre of methanol LR.
5. Nickel Nitrate Buffer Solution:
Dissolve 500 5 gm of Nickel Nitrate LR grade along with 21  1 ml
concentrated Nitric acid. Make the total volume to 420 ml with DM water.
6. Standardization of Mercuric Nitrate :
Take 25 ml of 0.02N Sodium Chloride solution in a Conical flask add 1 ml
Diphenyl carbozone and 1 ml Nickel nitrate buffer. Titrate against Mercuric
nitrate until the solution turns from green to Violet/bluish.
Normality of Mercuric Nitrate = 25 X 0.02/ Titre value of Mercuric nitrate.

Testing Procedure :

1. Take suitable aliquot of the sample add DiphenylCarbozone indicator

solution and Nickel Nitrate buffer solution (each 1 ml for every 50ml of
sample).
2. Titrate with 0.02N Hg (NO3)2 till colour of solution changes from green to
violet.
3. Take burette reading (BR) and calculate the chloride contents from
formula given below:
Chloride ppm as CaCO3 = TV X Normality of Hg (NO3)2 X1000 X50
Sample volume

Cooling Water Parameters 20 Analytical Procedures

13. B) Determination of Chloride by Silver Nitrate Method (Ref- IS 3025)

Reagents:

1. Potassium chromate (5%):

Dissolve 5 g of potassium chromate in 100 ml distilled water.

2. Silver nitrate (N/50):

Dissolve 3.4 g Silver Nitrate in 1000 ml of Distilled water.

3. Nitric acid (0.10%):

Dilute 0.1ml HNO3 in 100 ml Distilled water.

4. Sodium hydroxide (0.1%):

Dissolve 0.1 g NaOH in 100 ml distilled water.

5. Standardization of Silver nitrate :

Take 25ml of Sodium chloride 0.02N Solution in a conical flask add 0.5 ml
Potassium chromate indicator and titrate with Silver nitrate until solution
changes to brick red.

Normality of Silver Nitrate = 25 X 0.02/ Titre value of Silver nitrate.

Procedure:

100 cc water sample is taken in a conical flask (250-ml), add phenolphthalein

indicator. Neutralize with HNO3 or caustic. Add 2 drops of Potassium
chromate solution.Titrate against N/50 Silver nitrate until color is changed
from yellow to brickred. Run blank also.

Calculation:

Chloride as Cl, mg/l = (A - B ) N 35.45

ml sample

Where, N = Strength of silver nitrate

A = ml AgNO3 used for sample.
B = ml AgNO3 used for distilled water.

Cooling Water Parameters 21 Analytical Procedures

14. Sulphate Ion (SO4 --) by Turbidity method (Ref- APHA 4500 E)

Summary:
In this, sulphate ions in water to be tested are estimated by turbidity method.

Interference:
If sample is colored, make color blank.

Reagents:

1. Standard sulphate solution :

Dissolve 0.1479 gm anhydrous sodium sulphate in distilled water and dilute to
1000ml.
2. Buffer Solution A :
Dissolve 30 gm of magnesium chloride MgCl 2.6H2O, 5 gm of sodium acetate
CH3COONa.3H2O, 1 g KNO3 and 20ml acetic acid in 1litre distilled water.
3. Buffer solution B: (Required when the sample contains less than 10ppm
sulphate ions.)
Dissolve 30 gm of magnesium chloride MgCl2.6H2O, 5 gm of sodium acetate
CH3COONa.3H2O, 1 gm KNO3, 0.111 gm of sodium sulphate anhydrous and
20ml acetic acid in1 litre distilled water.
4. Barium Chloride :
Barium chloride crystals of mesh size 20 – 30.

Procedure:

Calibration and Analysis:

Take 2, 4, 6, 8, 10 ppm standard sulphate ion solutions in 100ml standard flasks.

To each flask add 25ml. of buffer solution add 0.5gm of barium chloride crystals
and measure absorbance at 420nm after 15 min diluting it to 100ml. Prepare
calibration curve of SO4 in ppm Vs Abs.

Repeat the above procedure using suitable aliquot of sample instead of standard
sulphate solution. Measure absorbance at 420nm and find out content of ions
from standard graph. Run blank simultaneously.

Calculation
Sulphates as SO4 = (Sample value-Blank value) X total volume/sample volume.

Cooling Water Parameters 22 Analytical Procedures

15. A) Dissolved Silica (Ref- APHA 4500 Si-E)

Summary:

Silica estimated by applying the absorbance method is only the silica that is
present in a dissolved state, and the silica in its comparatively stabilized granular
state. Silica in colloid form or if absorbed to other matter cannot be estimated as
such. It is to be converted into soluble or reactive silica state . The method given
here is applicable for reactive silica only.

In this method, the coloration of molybdenum yellow produced by the action of

dissolved silica upon ammonium molybdate is changed to molybdenum blue by
applying 1-amino, 2-napthol, 4-sulfonic acid, and then measuring its absorbance.

Apparatus required:
1. Spectrophotometer

Reagents:

1. 1:1 HCl

2. 10% Ammonium molybdate:

Dissolve 10 grams of ammonium molybdate approximately in 100 ml of
DM distilled water

3. 10 % oxalic acid solution :

Dissolve 100 g Oxalic Acid in 1000 ml Distilled water. Ensure complete
dissolution.

4. Reducing agent:
(A) 30 grams of sodium metabisulphite is dissolved in approximately 150
ml of DM distilled water
(B) Grind 0.5 grams of 1-amino, 2-napthal 4 -sulphonic acid with 1.0 gm of
Sodium Sulphite and dissolve in 50 ml DM water.

Solution (A) and (B) are mixed together

Cooling Water Parameters 23 Analytical Procedures

 Procedure:

1. Take 25 ml of sample in PVC cylinder and make it up to 50 ml.

2. Then add 1 ml 1:1 HCl, 2 ml Ammonium Molybdate solution, Wait for 5
minutes.
3. Then add 1.5 ml Oxalic Acid & 2.0 ml reducing reagent.
4. The silica content is analyzed on spectrophotometer, at 650nm wavelength
after 10 minutes. Run blank also.

Calculation

Dissolved Silica as SiO2 = (Sample value-Blank value) X total volume/sample

volume.

Calibration Procedure:

(A) Dissolve 3.13 gm of Sodium fluorosilicate in 1 litre of DM water, it

is equivalent to 1000 ppm of SiO2.
(B) Dilute 10 ml solution (from A ) to100 ml with DM Water. It is
equivalentto100 ppm of SiO2

Take 0,0.5,1, 2 ,3 ,4,5 ml From solution B separately in to a 100 ml PVC

measuring cylinders and make up the volume to 50 ml with distilled water. Then
add 1 ml 1:1 HCl & 2 ml Ammonium Molybdate solution. Wait for 5 minutes.
Then add 1.5 ml Oxalic Acid & 2 ml reducing reagent.

The silica content is analyzed on spectrophotometer, at 650nm wavelength after

10 Minutes.

Draw calibration graph (Absorbance Vs SiO2 Conc. in ppm).

This procedure follows Beer - Lambert's law.

Cooling Water Parameters 24 Analytical Procedures

 B) Total Silica Measurement Non-Reactive Silica

Principle:
Non-reactive silica can be converted to reactive silica by treating the sample
above, with Boric acid and Hydrofluoric acid and then the absorbance of the
solution is measured in a spectrophotometer.

Reagents Required:
a. 5% (W/V) Boric acid.
b. 48% Hydrofluoric acid (with very low flurosilicic acid)
c. 555 ml. of H2SO4 diluted in 1.5 litres of double distilled silica free water.
d. Hydrocholoric Acid (1:1)
e. 10% (W/V) Ammonium molybdate solution.
f. 10% (W/V) Oxalic acid.
g. Reducing Agent (A.N.S.A.)

Note: All the reagents are to be of AR grade.

Double distilled silica free water is to be used.
All containers used are to be of polyethylene.

Procedure:
Add to the 25 ml of stock solution, 50 ml of 5% Boric Acid in a 150 ml
polyethylene breaker. Add 0.5ml of 48% Hydrofluoric acid. Mix well and keep
the beaker in a water bath at 50oC for 10 minutes to drive away the excess HF.
Then add 4 ml of 10% Ammonium molybdate, and agitate. Allow the solution to
stand for 20 minutes. Maintain the temperature of the water bath at around 30 to
40oC.

Remove the beaker from the water bath. Add 20ml of H2SO4, with rapid stirring.
Allow the mixture to stand for 5 minutes.

Add 1 ml of Reducing agent, and allow the solution to stand for 20minutes or until
the solution attains room temperature, which ever is earlier. Measure the
absorbance of the solution on spectrophotometer at 815 nm. Obtain the value by
referring to the calibration curve. The value obtained is Total Silica. Total Silica
minus the Reactive silica will give the non-reactive silica.

Colloidal Silica (Non-Reactive Silica) = Total Silica - Reactive Silica

Cooling Water Parameters 25 Analytical Procedures

16. Oil and Grease (Ref- APHA 5520 D)

Reagents:

1. Sulphuric acid [1:1] :

Add slowly and carefully 250 ml conc. sulfuric acid to 250 ml distilled water and
cool.

2. Petroleum ether :

Procedure:

1. Place 1 liter or appropriate volume of the sample in a suitable separating funnel.

2. Add 5 ml sulphuric acid per liter of sample.
3. Rinse the sample bottle with 15 ml petroleum ether. Add further 25 ml ether to
the funnel and shake vigorously for 5 minutes.
4. Draw the aqueous phase in to a clear container and transfer the ether layer
through Whatman filter paper no 40 in to a clean previously weighed 100 ml
beaker.
5. Continue the extraction twice and add the ether extracts to the beaker.
6. Distill off the ether over a water bath.
7. Cool in a desiccator and weigh.

Calculations:

Residue in the beaker (mg) X 1000

Oil and grease (ppm) = ------------------------------------------------
Sample taken (ml)

Note: The method does not determine volatile fraction.

Cooling Water Parameters 26 Analytical Procedures

17. Total Suspended Solids (Ref- APHA 2540 D)

Apparatus:

1. Gooch crucible
2. Glass fiber filter paper GFD (Whatman)
3. Filtration assembly

Procedure:

1. Place the fiber filter paper on the gooch crucible and record the weight as W1.
2. Shake the sample well and filter a known volume of the sample through the
crucible containing the filter paper.
3. Wash the crucible with distilled water. Dry the crucible for 2 hrs. at 105 oC in a
oven. Cool in a desiccator and record the weight of the crucible as W2.

Calculation:

(W2 –W1) X 1000

Total Suspended Solids = -------------------------------
100
Where
W1 = Weight of empty crucible + Filter paper
W2 = Weight of crucible + filter paper + suspended solids after drying.

Cooling Water Parameters 27 Analytical Procedures

18. Total Dissolved Solids (Ref- APHA 2540 C)

Apparatus :

1. Evaporating dish
2. Glass fiber filter paper GF-A (Whatman 42)
3. Filtration assembly

Procedure:

1. Place the fiber filter paper on the gooch crucible / Whatman 42 filter paper in a
glass funnel and wet it with the sample.
2. Record weight of an empty evaporating dish as W1.
3. Filter the sample through the crucible and then measure 100ml sample and
transfer it to the evaporating dish and evaporate to dryness over a water bath.
4. Dry the evaporating dish in an oven at 105oC for 2hrs.
5. Record weight of evaporating dish with solids as W2.

Calculation :

Total Dissolved solids (ppm) = (W2 - W1) X 1000

100
W1 = weight of empty evaporating dish
W2 = weight of evaporating dish + residue

19. Measurement of pH

Procedure:

1. Rinse the electrode with MB water.

2. Dry the electrode with tissue paper.
3. Insert the electrode into the sample to be tested in a clean beaker.
4. Measure the pH by using appropriate mode. Constant reading obtained is
taken as the final reading.
5. Clean the electrode in MB water.
6. Keep the electrode immersed in 0.01N HCl, when not in use.

Cooling Water Parameters 28 Analytical Procedures

20. Conductivity (Ref- APHA 2510 B)

Procedure :
1. Take sample in a Clean Beaker.
2. Dip Conductivity Cell in the sample.
3. Wait for Stable Reading.
4. Record the Value.

21. Temperature

Temperature measurements are usually made with mercury filled centigrade

thermometer. The reading should be made by dipping the thermometer in the
sample. Reading to be taken after giving sufficient time and upon obtaining
constant reading. Normally temperature should be read at the time of sampling.
The temperature should be expressed to the nearest degree centigrade.

22. Free residual Chlorine

Free residual chlorine is normally determined at site using test kits. Chlorotex
reagent is widely used. Other kits which have wide acceptance are kits using Ortho-
toludine reagent or DPD reagent kits. In the laboratory the DPD titrometric method
is extensively used. Test procedures are given along with the test kits and the same
to be adopted. For Qualigen’s chlorotex test, for every 9.0 ml sample, 1.0 ml of
reagent is to be used and mixed gently. Wait for two minutes for the appearance of
color. Compare it with the color comparator affixed to the bottle to know the value
of chlorine in ppm.

Test Kit Procedure

For determination of Free Chlorine up to 4 ppm in water.

Fill the water sample for analysis in the Easy test jar up to the 10ml mark. Add 1
micro spoon of Reagent-A. Shake well.

If pink color does not appear free chlorine is nil. If pink color appears, add Reagent
– B drop by drop till the pink color is discharged. Shake the jar after the addition of
each drop.

Count the number of drops added to the above solution.

Free residual chlorine in ppm = No of drops consumed X 0.1

Note: For more details see Booklet which is provided along with the Test Kit.

Cooling Water Parameters 29 Analytical Procedures

23. Chemical Oxygen Demand (Ref- APHA 5220 B)

Reagents :

1. Standard potassium dichromate digestion solution - 0.0417 M

(0.25N):
Dissolve 12.259 gm of K2Cr2O7 ( previously dried at 1030C for 2hrs.) in
500ml distilled water and dilute to 1000ml.
2. Sulphuric acid reagent :
Add Ag2SO4 (silver sulfate) powder to the standard sulphuric acid at the rate
of 5.5 gm/Kg H2SO4. Let stand for a day to dissolve Ag2SO4.
3. Ferroin indicator Solution :
Dissolve 1.485 gm of 1,10-phenanthroline mono hydrate and 695mg
FeSO4.7H2O in distilled water and dilute to 100ml.
4. Standard Ferrous Ammonium Sulphate FAS ( 0.1 N ) :
Dissolve 39.2 gm Fe(NH4).2(SO4)2.6H2O in distilled water. Add 20 ml
sulphuric acid, cool and dilute to 1000ml. Standardize against 0.25N
K2Cr2O7
5. Mercuric sulphate :
6. Sulfamicacid :
Required only to remove nitrite interference.

Procedure :

Take 5 ml of sample in a 100ml round bottom flask add 0.4 gm of mercuric

sulphate and 10 ml of K2Cr2O7 and 15ml distilled water and 30 ml of H2SO4 and
keep for reflux for 2 hours. After cooling add 90 ml of distilled water, add 3 drops
of ferroin indicator and titrate with 0.1N FAS. Run blank using distilled water in
place of sample.

Calculation :

COD ( ppm ) = (Blank – titer value) X Normality of FAS X 8000

Sample Volume (ml)

Cooling Water Parameters 30 Analytical Procedures

24. Nitrites (Ref- Modified For Residue)

Reagents:

1. CAS (Cerci Ammonium sulphate 0.1 N)

2. Ferroin Indicator

Procedure:

Take 50 ml sample in a conical flask. Add 3 drops of Ferroin indicator. Titrate with
0.1N Cerci Ammonium sulphate solution. The end point is from orange red to pale
blue or yellowish green.

Calculations:

Nitrites as NaNO2 = 34.5 X N X T.V X 1000

V

N = Normality of CAS
V = Volume of Sample(ml)

Cooling Water Parameters 31 Analytical Procedures

25. Nitrates (Ref- NPDES & SDWA)

Reagents:

Sulfuric acid solution:

Carefully and slowly add 500ml of conc. sulfuric acid to 75ml of distilled water.
Cool and store in a bottle.

Brucine sulphanillic acid solution :

Dissolve 1g of brucine sulfate and 0.1gm of sulphanalic acid in 70ml of hot distilled
water. Add 3ml of conc. hydrochloric acid cool and dilute to 100ml, with distilled
water.

Sodium Arsenite solution:

Dissolve 1.83g of sodium arsenite (NaAsO2) in 1liter of distilled water.

Potassium nitrate Standard solution:

Dissolve 1.631 potassium nitrate in 40ml of distilled water and dilute to one liter.
1ml = 1mg nitrate

Calibration:

1. Prepare a blank and series of standards using the standard nitrate solution.
The standards should cover the range from 0 - 50 ppm as NO3.
2. In to a clean dry 50ml beaker pipette out 5ml of the standard add 1ml of
Brucine sulphanalic acid regent in to a second 50 ml beaker, 10ml of sulphuric
acid.
3. Mix the contents of the beaker carefully adding the sample with brucine acid
reagent to the beaker containing the acid. Pour from one beaker to the
another six times to ensure mixing.
4. Set aside for 10 min in a dark place.
5. While the color is developing add 10 ml of distilled water to the empty 50ml
beaker. After 10 min interval add to the 10ml distilled water to the sample
and mix as before. Allow to cool for 20 - 30 minutes in a dark place.
6. Measure the absorbance of the above standards at 410 nm. Prepare reagent
blank using DM distilled water and set the instrument to zero absorbance
using the same.
7. Plot a graph of absorbance Vs the Nitrate concentration in ppm.

Cooling Water Parameters 32 Analytical Procedures

 Procedure:

1. In to a clean dry 50 ml beaker, pipette out 5ml portion of the sample.

2. Add 1ml of brucine sulphanalic acid reagent.
3. In to a second beaker measure 10 ml of sulfuric acid.
4. Mix the contents of the two beaker by carefully adding the sample with the
brucine sulphanalic acid reagent to the beaker containing acid. Pour from one
beaker to another six times to ensure mixing.
5. Set aside for 10 min in a dark place.
6. While the color is developing add 10 ml of distilled water to the empty 50 ml
beaker.
7. After 10 min interval add to the 10ml distilled water to the sample and mix as
before. Allow to cool for 20-30 min in a dark place.
8. Measure the absorbance of the sample at 410nm. Prepare a reagent blank
using DM distilled water and set the instrument to zero absorbance using this
solution.
9. Determine the concentration of the sample from the graph.

NOTE :

If residual chlorine is present add 0.1ml of arsenite solution to 50ml. portion of

sample for each 0.05ppm of chlorine, add a drop in excess and mix.

Calculation:

Find out nitrate content in ppm from graph.

Cooling Water Parameters 33 Analytical Procedures

26. KMnO4 Value (Ref- Physico-Chemical Examination of Water, Sewage and
Industrial Effluents by N.MANIVASAKAM 2nd Edition (pg. no. 200 /204 ) )

Reagents:
1) 0.125 N KMnO4 stock solution:
Dissolve 4 -4.2 g of KMnO4 in about 900ml distilled water to make a final
volume of 1L Standardize against 0.125 N sodium oxalate solution.

2) 0.125N Sodium oxalate:

Dissolve 2.0938 g sodium oxalate in distilled water & make final volume to
250ml. This is exactly 0.1250 N Place 25ml of this solution in 500 ml conical
flask and add 150 ml 2 N sulfuric acid (Prepared by mixing 50 ml conc.H 2SO4
and 950 ml distilled water). Titrate with the potassium Permanganate solution
rapidly at room temp. to the persistence of a faint pink color. Allow to stand
until the solution becomes color less. Then heat to 55-600c and complete the
titration until a faint pink color persists for 30 seconds.

3) 0.0125 N KMnO4 solution:

Dilute appropriate volume of 0.125 N KMnO4 solution ten times such that the
resulting solution is 0.0125 N KmnO4.

1ml of 0.0125 N KMnO4 = 0.1 mg of oxygen.

4) H2SO4 (1+3): Add carefully 250ml con H2SO4 into 750ml MB water with
constant stirring and cool.
5) Potassium iodide (10%): Dissolve 10g potassium iodide and dilute to
100mlL store it in an amber colored bottle.
6) Starch indicator: Dissolve 1g starch in 100ml of boiling MB water and
cool.
7) Na2S2O3 solution 0.125N: Dissolve 31.03 g Na2S2O3.5H2O in MB water
and dilute to 1L. Standardize it against 0.125 N K2Cr2O7 solution.

8) Na2S2O3 solution 0.0125 N: Dilute an appropriate volume of 0.125 N

Na2S2O3 ten times such that the resulting solution is 0.0125 N Na2S2O3.

9) K2Cr2O7 solution: Dissolve 6.13 g K2Cr2O7, (previously dried at 1050C for

2 hrs) in distilled water and make up to 1000 ml. in a volumetric flask. This is

Cooling Water Parameters 34 Analytical Procedures

 exactly 0.1250 N solution. Place 100 ml boiled & cooled distilled water in a 500 ml
conical flask. Add 3 g potassium iodide, 2.0 g sodium bicarbonate and shake until
the salts dissolve. Add 6 ml conc. HCl and pipette 25.0 ml. 0.1250 N potassium
dichromate solution in to the flask. Cover the flask with a watch glass and keep it in
dark for 5 minutes. Rinse the watch glass and dilute the solution in the flask to 250
ml with boiled & cooled distilled water, titrate the liberated iodine with the sodium
thiosulfate solution in the burette. When the solution acquires yellowish green
color, add 1 ml. starch indicator solution. Now a blue shade will be obtained. Rinse
the sides of the flask and continue the titration until one drop changes the color
from greenish blue to light green.

2. Procedure:
1) Place 250 ml of the well mixed sample in a stopper glass bottle. Add 10 ml
of sulfuric acid (1+3) followed by 10 ml of 0.0125 N KmnO4 (the volume of KMnO4
added may be such that half of it remains after 4 hours). Accordingly, 20 ml or 30
ml may be added depending upon the nature of sample. Prepare a blank also in
another conical flask.
2) Keep all the samples in the dark for 4 hours.
3) At the end of 4 hours, add 1 ml of KI and titrate with 0.0125 N N2S2O3
using starch indicator. End point is blue to colorless.

3. Calculation:
(Blank - Sample) x 0.1 x 1000_
Oxygen absorbed (at 270 C, 4 hrs) ppm = sample (ml.) taken for determination

4. Notes:
The amount of residual chlorine in treated waters does not interfere with this test
but however excess amounts of residual chlorine interfere and hence the sample
should be suitably de-chlorinated with sodium sulfate before the test is carried out
Inorganic substances such as nitrite, sulfide, ferrous iron react with KmnO 4.

Cooling Water Parameters 35 Analytical Procedures

27. Ammonical Nitrogen For ETP (Ref- APHA 4500 Norg B)

Reagents:

Borate Buffer Solution:

Add 88 ml 0.1N NaoH to 500 ml approximately 0.025M Sodium tetraborate (Na2B4O7)

Solution (9.5 grams Sodium tetra borate and 88 ml 0.1 N NaoH Solution Dilute to 1 liter.

Mixed Indicator Solution:

Dissolve 0.2 grams Methyl red indicator in 100 ml 95% ethyl or isopropyl alcohol.
Dissolve 0.1 grams methylene blue in 50 ml 95% ethyl or isopropyl alcohol.

Indicating boric acid Solution:

20 grams Boric acid in Ammonia free distilled water, add 10 ml mixed indicator Solution
,add dilute to 1 liter prepare monthly.

Procedure :

Take 500 ml Sample to a distillation flask and add 25 ml borate buffer Solution and
adjust pH 9.5 with 6 N NaOH .Distill & collect 200 ml distillate .Use 50 ml indicating boric
acid as absorbent solution.

Titrate ammonia in distillate with Standard 0.02 N H2SO4 titrant until indicator turns a
pale lavender.

Caculation :

Titer value of H2SO4 X Normality of H2SO4 X 1000 X 14

Ammonical Nitrogen = -------------------------------------------------------------------

( ppm ) Sample Volume(ml)

Cooling Water Parameters 36 Analytical Procedures

28. Determination of PO4 in Water by SnCl2 method (Ref- APHA - 4500 P. D /
AQUAPHARM PROCEDURE)

Scope:

This method covers the determination of ortho-phosphate, Inorganic phosphate, Organic

phosphate and total phosphate in water. This method is applicable for Steam drum B/D,
SHP steam, PO4 tank, and other water Samples from complex.

Summary:

For ortho- phosphate, an known volume of sample is treated with ammonium molybdate
and SnCl2 The blue colour formed is measured with spectrophotometer. For total
inorganic phosphate an known volume of sample is digested with Sulfuric acid and for
total phosphate an known volume of sample is digested with H2SO4 and ammonium per-
sulfate and then phosphate is estimated by developing the colour as per procedure of O-
phosphate. Organic phosphate is the difference between total phosphate and total
inorganic phosphate.

Principle:

Ammonium molybdate in acidic medium reacts with ortho-Phosphate to form

molybdophosphoric acid which is reduced by stannous chloride to intensely coloured
molybdenum blue. Poly-phosphates are converted to Ortho-phosphate by acid
hydrolysis. Organo-phosphorous compounds are converted to orthophosphate by
treating with H2SO4 and Ammonium persulphate.

Interference:

Coloured and hazy sample may alter the result.

Instruments / Equipments / Glassware / Materials:

Instruments / Equipments:

a) UV/VIS Spectrophotometer

b) Hot plate

Cooling Water Parameters 37 Analytical Procedures

Glassware / Materials:

a) Laboratory glass wares

b) Quartz/Glass cell: 10 mm or 20 mm path length

Chemicals/Solutions:

Chemicals:

a) Sulphuric Acid: AR Grade or equivalent.

b) Ammonium Persulphate: AR Grade or equivalent.

c) Stannous chloride: AR Grade or equivalent.

d) Phenolphthalein indicator: AR Grade or equivalent.

e) Ammonium molybdate: AR Grade or equivalent.

f) Potassium dihydrogen phosphate: AR Grade or equivalent.

g) Sodium hydroxide: AR Grade or equivalent

Solutions:

a) Stock solution of Phosphate

b) 2.5% Stannous Chloride solution

c) Sulphuric Acid 1:1 Solution

d) Phenolphthalein indicator

e) Ammonium molybdate Soln.

f) 4 N Sodium hydroxide solution

g) 2 N Sulphuric acid solution

Safety:

Normal laboratory safety practices shall be followed. Digestion of the sample should be
done in well ventilated fume hood.

Sampling Procedure Reference:

Samples are provided by the respective plant operators to the wet laboratory.

Cooling Water Parameters 38 Analytical Procedures

Calibration:

a) Prepare Std. Phosphate solution (1ml = 0.01 mg of PO4) by diluting 10 ml of stock

solution of Phosphate (1ml = 1mg) to 1000 ml in DM water.

b) Pipette out 2, 4, 6, 8 & 10 ml of standard phosphate solution (1ml=0.01 mg PO4) in

50 ml vol. flask / Nesseler tubes. Cary out a blank by omitting standard in another 50 ml
vol. flask / Nesseler tube.

c) Dilute each solution to about 10 ml with DM water & mix.

d) Add 2 ml of ammonium molybdate solution to each flask and wait for five minutes.

e) Add 0.5 ml of stannous chloride solution to each flask and mix. Make up to 50ml with
DM Water and mix well.

f) After 10 minutes measure the absorbance at 690 nm wavelength with reference to

blank as zero. (Cell length =10 mm)

g) Plot a curve of absorbance v/s conc. in mg of phosphate and find out slope value or
calculate slope value as per given formula

Concentration in mg of PO4

Slope = ------------------------------------

Absorbance reading

Analytical Procedure:

O-phosphate:

a) Pipette out 5 ml sample or a suitable aliquot to 100 ml vol. flask/ Nesseler tube.
Dilute to about 40 ml with DM water. Carry out the blank by omitting sample in to
another volumetric flask/Nesseler tube.

Note:

i) For the lower concentration level of Phosphate, take 200 ml of sample and digest to
10-20 ml on hot plate & transfer quantitatively in 50 ml volumetric flask.

ii) For Phosphate tank sample, take 2 ml sample and dilute it to 1000 ml with DM water.
From this take 1 ml sample for the test.

Cooling Water Parameters 39 Analytical Procedures

b) Add 4 ml ammonium molybdate reagent, wait for five minutes and add 0.5 ml
Stannous Chloride solution.

c) Make up to 100 ml with DM water and mix well.

d) Measure absorbance after 10 minutes of the Sample using Spectrophotometer against

reagent blank at 690 nm with l0 mm or 20 mm cell.

e) Note down the absorbance reading.

Total Inorganic phosphate:

a) Transfer 5 ml Sample or a suitable aliquot to 250 ml beaker. Add 1 ml 1:1 Sulfuric

acid reagent. Dilute to about 100 ml with DM water. Carry out blank by omitting sample
into another beaker in a same manner.

b) Keep the beaker on hot plate and boil until the volume is reduced to 10-20 ml.

c) Cool and neutralize to faint pink colour with 4N NaOH solution by using
phenolphthalein as an indicator.

d) Neutralize it again by adding 2N H2SO4 till pink colour just disappears.

e) Quantitatively transfer it to 100 ml volumetric flask or Nesseler tube. Rinse the beaker
twice with 5 ml DM water each time and add to the vol. flask.

f) Proceed as per step b) to d) of above Sec.

g) Note down the absorbance reading.

Total Phosphate:

a) Transfer 5 ml Sample or a suitable aliquot to 250 ml beaker. Add 1 ml 1:1 Sulfuric

acid reagent and 0.4 gm of Ammonium persulphate. Dilute to about 100 ml with DM
water. Carry out blank by omitting sample into another beaker in a same manner.

b) Proceed as per step b) to f) of above Sec.

c) Note down the absorbance reading.

Cooling Water Parameters 40 Analytical Procedures

Calculation:

a) Calculate O - Phosphate, Total Inorganic Phosphate & Total Phosphate as mg/litre PO4
using respective absorbance readings by given formula:

Absorbance reading x 1000 x Slope value

PO4 mg/litre = -----------------------------------------------------

Sample vol. (ml)

b) Organic phosphate = Total phosphate - Total Inorganic phosphate

c) Phosphate as P = PO4 (mg/l) x 0.33

Note: Take the slope value according to the Cell path length (10 or 20mm) used for the
test and apply dilution factor if sample is diluted.

Report:

Report the results as given bellow

Up to 100 ppb- Multiple of 5 ppb

0.1 to 1 ppm– 1st decimal

1 to 15 ppm- 1st decimal with multiple of 0.5 ppm

Above 15 ppm- Whole number

Cooling Water Parameters 41 Analytical Procedures

29. Determination of Sulphide (Ref- IS – 3025, Part 29)

Scope:
Titrimetric iodine method (Iodometric) for the determination of total and dissolved
sulphides in waters and wastewaters is suitable for measurement of sulphide in
concentration above 1 mg/l, if interferences are absent and loss of hydrogen sulphide is
avoided.

Iodometric Method

Principle and Theory:

Sulphides are stripped from the acidified sample with an inert gas and collected in zinc
acetate solution. Excess iodine solution, added to the zinc sulphide suspension, reacts
with the sulphide under acidic condition. Thiosulphate is used to measure un reacted
iodine to indicate the quantity of iodine consumed by sulphide.
The reaction may be given as follows:

1) S + I2 = S2 + 2 I

2) I2 (excess) + 2 S203 = S406 + 2 I

Interferences:

Reduced sulphur compounds, such as sulphite, thiosulphate and hydrosulphite, which

decompose in acid, may yield erratic results. Volatile iodine consuming substances will
give high results.
Eliminate interferences due to sulphite, thiosulphate, iodide and many other soluble
substances, (but not Ferro-cyanide) by first precipitating zinc sulphide, removing the
supernatant, and replacing it with distilled water. Use the same procedure, even when
not needed for removal of interferences, to concentrate sulphide.

Procedure:

Put required quantity of 2 N zinc acetate solution into 500 ml glass bottle. Fill with
sample and add required quantity of 6 N sodium hydroxide solutions. Put stopper with no
air bubbles under stopper and mix by rotating back and forth vigorously about a
transverse axis. Addition of reagents may be varied in volume so that the resulting
precipitate is not excessively bulky and settles rapidly. Add enough sodium hydroxide to
produce a pH above 9. Let the precipitate settle for 30 minutes. Filter the precipitate
through glass fiber filter paper and carryout titration immediately.

Sampling and Storage:

Sampling and storage shall be done as prescribed in IS: 3025 (Part I) 1986 'Methods of
sampling and test (physical and chemical) for water and wastewater: Part 1 Sampling
(first revision)',
Samples must be taken with a minimum of aeration and preserved at low temperature
(Sulphide may be volatilized by aeration and any oxygen is advertently added to the
sample may convert the sulphide to an immeasurable form).

Cooling Water Parameters 42 Analytical Procedures

Preserve the sample with addition of 2 mill of zinc acetate. Samples not preserved must
be analyzed immediately.

Apparatus:

Reaction flask- Wide mouth bottle of 1 liter capacity, with a 2 hole stopper, fitted with a
fritted gas-diffusion tube (plastic, ceramic or glass and a gas outlet tube).
Absorption flasks - Two 250-ml capacity long necked flask with 2 hole stoppers fitted
with glass tubes and suitable connections to pass gas through in series.

Reagents:

Zinc acetate solution (2 N) - Dissolve 110 gm Zn (C2H3O2)2.2H20 in 400 ml distilled

water and finally make up to 1 liter.

Inert gas - A cylinder of nitrogen [pure grade, see IS: 1747-1972 Specification for
nitrogen (first revision)] or CO2 or a CO2 gas generator [Grade 1, see IS: 307~1966
Specifications for carbon dioxide (second revision)].

Sulphuric acid concentrated

Standard iodine solution (0.025 N) - Dissolve 20-25 gm potassium iodide (KI) in a little
water and add 3.175 gm iodine. After iodine has dissolved, dilute to 1 liter with distilled
water, standardize this solution against 0.025 N sodium thiosulphate using starch
indicator.

Hydrochloric acid concentrated

Standard thiosulphate solution (0.025 N) - Dissolve 6.205 gm Na2S203.5H20 in 800 ml

boiled and cooled distilled water. Add 0.4 gm NaOH or 5 ml chloroform as a preservative
and finally make up to 1 liter.

Starch indicator solution - Add 5.0 gm starch to 800 ml boiling distilled water and stir.
Dilute to one liter and boil for few minutes and let settle over night. Use the clear
supernatant. (This solution may be preserved by adding 1.25 g salicylic acid/liter or by
adding a few drops of toluene).

Aluminium chloride solution (6 N) - Take the 100 gm AlCl3.6H20 from a previously

unopened reagent bottle and dissolve in 144 ml distilled water.
Note - Because of the hygroscopic and caking tendencies of this chemical, it will be
convenient to purchase in small packing.

Sodium hydroxide (6 N) - Dissolve 240 gm NaOH in distilled water and dilute to 1 liter.

Procedure:

Total sulphide
Take 5 ml zinc acetate solution and 95 ml distilled water into each of the two absorption
flasks.
Connect the reaction flask and two absorption flasks in series and purge the system with
CO2 or N2 for 2 minutes. Measure 500 ml well mixed sample into the reaction flask.

Cooling Water Parameters 43 Analytical Procedures

Acidify the sample with 10 ml concentrated H2S04 and replace the prepared 2 holes
stopper tightly; pass N2 or CO2 ( not air or oxygen) through the sample for 1 hour or
until the experiments show no more sulphide coming over.
To each of the absorption flasks, then add iodine solution well in excess of the amount
necessary to react with the collected sulphide.
Add 2.5 ml concentrated HCl acid to each flask, stopper and shake to mix thoroughly.

& Transfer contents of both flasks and back titrate with 0.025 N sodium thiosulphate
solution using starch solution as indicator. Run a blank parallel for accurate results.

Dissolved Sulphide
Remove suspended solids in the sample by flocculation and settling.
Fill 1 liter bottle with flowing sample in such a way that the sample, which has had the
least possible contact with air. Add 2 ml aluminium chloride solution and 2 ml NaOH
solution and stopper with no air bubbles under the stopper. Rotate back and forth about
a transverse axis as vigorously as possible for at least 1 minute in order to flocculate the
contents thoroughly.

Note - The volume of these chemicals may be varied according to experience, the idea
being to get good clarification without using excessively large amounts.

Allow to settle for 15 minutes, or until supernatant liquid is reasonably clear.

Alternatively remove suspended matter by centrifugation.

Proceed as for total sulphide after taking 500 ml sample into the reaction flask.

Calculation:

(V1-V2)*400
mg/l, Sulphide = --------------------------------
V

Where
V1 = volume in ml of standard iodine solution added,
V2 = volume in ml of standard thiosulphate solution used.
V = volume in ml of sample taken.

Cooling Water Parameters 44 Analytical Procedures

 STANDARDISATION AND CALIBRATION OF INSTRUMENTS

1. TURBIDITY STANDARD PROCEDURE (Ref- APHA 2130 B)

Turbidity standard 4000NTU :

The particles of formazin are uniform in size and shape. The stock standards
prepared from formazin are accurate within 1% and stable for 6-8 months

Preparation:

Solution A:

Take 5 g of reagent grade “Hydrazine sulphate” and dissolve in 400ml of distilled

water.

Solution B:

Dissolve 50 g of pure “HexamethyleneTetramine” in 400ml of distilled water.

Mix solution A & B and make it up to one litre by adding distilled water and allow
this mixture to settle for 48 hours at normal room temperature.

This is stock solution of 4000 NTU strength of Formazin. This solution is usually
stable for a period of 6-8 months and working standard can be prepared as required
using following table.

Volume of stock solution (ml) to

Final strength of Formazin in diluted
make up 1 litre with higher grade
stock solution (NTU)
distilled water
125 500
25 100
12.5 50
20 of 500 NTU Solution 10
10 of 500 NTU Solution 5

Shake this stock solution well before dilution

Cooling Water Parameters 45 Analytical Procedures

2. pH METER CALIBRATION PROCEDURE (Ref- APHA 4500- H+B)

1. Method Of Calibration:

The instrument is calibrated by checking the pH of standard buffers.

2. Frequency Of Calibration:

pH meter is calibrated everyday during the beginning of the day.

3. Calibration Standard To Be Employed:

Dissolve one packet / Tablet of the buffer powder in 100 ml of MB water. Buffer
solutions should not be used beyond one month from the data of preparation.

4. Acceptance Criteria:

pH meter reading set to 7.00, then the other buffers viz,9.20 & 4.00 should
adjust to +/- 0.1 pH value off the standard value.

In case the variation lies between 0.1 – 0.2 correction factor of actual variation
is applied to the measured value.

In case the variation is > 0.2, the instrument is taken for service.

Note- Keep the electrode in 0.1 N HCl, when not in use

Cooling Water Parameters 46 Analytical Procedures

3. CONDUCTIVITY METER CALIBRATION PROCEDURE (Ref- APHA 2510 B)

Procedure :

1. Take sample in a Clean Beaker.

2. Dip Conductivity Cell in the sample.
3. Wait for Stable Reading.
4. Record the Value.

Calibration of Conductivity Meter :

The instrument is calibrated by checking the conductivity of standard KCl

solution daily

Concentration of KCl Solution Conductivity at 25° C

1.0 N 111.3 ms

0.1 N 12.85 ms

0.01 N 1.408 ms

0.001 N 146.9 ms

Preparation of 1N K Cl :

Dissolve 7.45 g of K Cl in 100 ml of distilled water and use for one month from the
date of manufacturing.

NOTE:

Keep the cell in distilled water, when not in use

Cooling Water Parameters 47 Analytical Procedures

4. ORP ELECTRODE CALIBRATION PROCEDURE

The performance of an ORP electrode can be determined by use of the ORP

Calibration Kit and the Procedures given below. The Calibration Kit consists of the
following items:
1. pH 4 Buffer sachet
2. pH 7 Buffer sachet
3. 100 mL conical flask
4. Plastic applicators
5. Indicator bottle
6. Instruction sheet

Procedure:
1. Take 100 mL of distilled water in a 250 mL conical flask.
2. Add the contents of the pouch labeled pH4 into it.
3. Dissolve it completely.
4. Measure the pH of the solution. It should be 4.0±0.1
5. Add a spatula full of the indicator powder in to the solution.
6. Stir it for 5 minutes. A small amount of indicator MUST remain un-dissolved. If
the entire indicator dissolves add a small amount and stir. Repeat as necessary
until a small amount of indicator remains un-dissolved.
7. Keep the ORP electrode in a beaker of distilled water for 5 minutes.
8. Rinse the ORP electrode and pat it dry with a soft tissue.
9. Put it in the beaker filled with pH 4 buffer/indicator solution.
10. Stir the electrode gently and let it rest against the side of the beaker.
11. Allow the reading to stabilize 30 to 60 seconds typically and note the reading.
12. The ORP value should be 482 mV Vs NHE. If the reference electrode is
Saturated Calomel electrode, then the ORP value is 242 mV.
13. The reading should be ±15 mV from the above value.
14. Repeat the above steps with pH 7 sachet.
15. The reading should be 285 mV (NHE) or 43 mV vs. SCE.

Buffer pH NHE* SSC* SCE*

4.0 482 285 246
7.0 285 88 49
*NHE : Normal Hydrogen Electrode, SSC : Silver - Silver chloride saturated with
NaCl and SCE : Saturated calomel electrode
The difference between the two values should be 197±20 mV.

Cooling Water Parameters 48 Analytical Procedures

5. WEIGHING BALANCE CALIBRATION PROCEDURE

Method of Calibration:

1. The balance is calibrated over complete range and checked for its performance
capability by manufacturer or servicing agent once in six months or when the
balance is found to be out of calibration. The standard and acceptance will be
specified by the servicing agent.

2. In addition the balance is calibrated after switching on everyday using 200 g

weight. This weight is checked once in six months by the servicing agent.

Acceptance Criteria:

0.3% variation in weight is accepted without any correction factor. For 0.31 to
1.00% variation the same correction factor is applied.

Cooling Water Parameters 49 Analytical Procedures

 MICROBIOLOGICAL EVALUATION METHODS

1. EVALUATION OF TBC/SRB - DIP SLIDE METHOD

TBC
Sterile plastic pieces coated with nutrient medium. Remove from container, rinse in
cooling water for 15-30 seconds, put back in container after draining excess water
from slide. Keep in warm place / Incubator for 48 Hrs.
Bacterial Colonies developed are counted using the comparator chart.

SRB
Sterile medium filled in sachets to be carefully cut open. Drain the contents
carefully into the sterile plastic container. Cooling water sample to be added into
the same container carefully / slowly without too many air bubbles
Fill the container to brim and leave it in a warm place. (Invert the container once to
ensure proper mixing of medium with the sample).
Periodically inspect the container for blackening of medium.

Interpretation - Blackening within

- 12Hrs - 106 orgs / 100ml
- 24 Hrs - 105 orgs / 100ml
- 48 Hrs - 104 orgs / 100ml
- 72 Hrs - 103orgs / 100ml
- 96 Hrs - 102 orgs / 100ml
- 120 Hrs - 101 orgs / 100ml
- 144 Hrs - not considered.

Cooling Water Parameters 50 Analytical Procedures