Method of Analysis - Water

DUBAI ELECTRICITY AND WATER AUTHORITY
G - STATION LABORATORY METHODS
LIST OF CONTENTS
GLAB - 01 Alkalinity, Caustic Page 1

GLAB - 02 Alkalinity, Carbonate, Bicarbonate and Hydroxide. Page 4
GLAB - 03 Ammonia Page 8
GLAB - 04 Ammonia in sea water and waste water Page 11
GLAB - 05 Atomic absorption/emission spectrometry Page 15
GLAB - 06 Biological oxygen demand (HACH method) Page 22
GLAB - 07 Biological oxygen demand (Winkler method) Page 27
GLAB - 08 Carbon dioxide, Free Page 33
GLAB - 09 Chemical oxygen demand (HACH method) Page 36
GLAB - 10 Chemical oxygen demand (Reflux method) Page 39
GLAB - 11 Chemical tank concentration (Refractometer method) Page 43
GLAB - 12 Chemical tank concentration (Sp. gravity bottle method) Page 46
GLAB - 13 Chemical tank concentration (Density balance method) Page 49
GLAB - 14 Chloride (Low range) Page 52
GLAB - 15 Chloride (Turbidity method, Very low range) Page 56
GLAB - 16 Chloride (High range) Page 59
GLAB - 17 Chlorine (Free and total) Page 62
GLAB - 18 Chlorine (Total available) Page 66
GLAB - 19 Chlorine dioxide (DPD method) Page 69
GLAB - 20 Chlorine dioxide (HACH method) Page 72
GLAB - 21 Chlorine demand Page 75
GLAB - 22 Chromate, Colourimetric Page 78
GLAB - 23 Coliform bacteria (Fecal) Page 81
GLAB - 24 Coliform bacteria (Total) Page 85
GLAB - 25 Coliform bacteria (MPN method) Page 90
GLAB - 26 Conductivity Page 97
GLAB - 27 Copper Page 100
i
LIST OF CONTENTS
GLAB - 28 Corrosion rate (Coupon method) Page 104

GLAB - 29 Corrosion rate (Probe method) Page 108
GLAB - 30 Dissolved oxygen (Indigo carmine method) Page 114
GLAB - 31 Dissolved oxygen (Modified Winkler method) Page 117
GLAB - 32 Dissolved oxygen (Test kit method) Page 120
GLAB - 33 Dissolved oxygen (Using meter) Page 122
GLAB - 34 Elimin-Ox (HACH method) Page 125
GLAB - 35 Ferric chloride (Iodometric) Page 128
GLAB - 36 Hardness, Total, Calcium, and Magnesium Page 131
GLAB - 37 Hydrazine (Colourimetric) Page 135
GLAB - 38 Hydrazine (Iodimetric) Page 138
GLAB - 39 Hydrochloric acid concentration % Page 141
GLAB - 40 Iodine number of Activated Carbon Page 144
GLAB - 41 Ion Chromatography, Anions Page 147
GLAB - 42 Ion selective electrode determinations Page 153
GLAB - 43 Iron (Phenanthroline method) Page 176
GLAB - 44 Iron (T.P.T.Z method) Page 179
GLAB - 45 Moisture content (At 105 °C) Page 182
GLAB - 46 Nitrite Page 184
GLAB - 47 Oil content Page 187
GLAB - 48 Ph Page 191
GLAB - 49 Phosphate, Ortho (High range)(Ascorbic acid reduction method) Page 194
GLAB - 50 Phosphate, Ortho (Low range)(Ascorbic acid reduction method) Page 197
GLAB - 51 Phosphate, Ortho (High range)(Vanadate method) Page 200
GLAB - 52 Phosphate ,Ortho (Stannous chloride method) Page 203
GLAB - 53 Phosphate (Ortho + meta)(Low range)(Ascorbic acid reduction) Page 206
GLAB - 54 Phosphonate (P205) Page 209
GLAB - 55 Polyphosphate based inhibitors Page 212
ii
LIST OF CONTENTS
GLAB - 56 Quality checks, acidity & alkalinity by autotitrator Page 2 15

GLAB - 57 Quality check of limestone Page 220
GLAB - 58 Salinity (Soils) Page 223
GLAB - 59 Salinity, Chlorosity, and Chlorinity (Water) Page 227
GLAB - 60 Settled volume Page 230
GLAB - 61 Silica (Low range) Page 232
GLAB - 62 Silt Density Index (SDI) Page 236
GLAB - 63 Sodium chlorite (Iodimetric) Page 241
GLAB - 64 Sodium sulphite (Iodimetric) Page 245
GLAB - 65 Specific gravity/Density of liquids (Density balance method) Page 248
GLAB - 66 Sulphate (Gravimetric) Page 251
GLAB - 67 Sulphate (Turbidimetnic) Page 254
GLAB - 68 Suspended solids Page 258
GLAB - 69 Tannin Page 261
GLAB - 70 Total dissolved solids at 180 °C Page 264
GLAB - 71 Total Kjeldahl Nitrogen Page 267
GLAB - 72 Total Nitrogen (HACH method) Page 271
GLAB - 73 Total Organic carbon, Total carbon, & Inorganic carbon Page 274
GLAB - 74 Turbidity (Nephlelometnic method) Page 279
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Page 1
G-STATION LABORATORY METHOD
SECTION: WATER
THE DETERMINATION OF CAUSTIC ALKALINITY
Method No. : GLAB - 01

Date issued : l3thMarch 1995
Date revised : 01st June 2008
Prepared by : K.P. Abraham
Approved by : Noora Bushawab
Page 2
DUBAI ELECTRICITY & WATER AUTHORITY Laboratory Methods of Analysis
TEST METHOD FOR WATER

CAUSTIC ALKALINITY
G-LAB METHOD No.01
(By K.P. Abraham)
1. SCOPE
This method covers the volumetric determination of free hydroxide ion in water and
wastewater. It is intended primarily for alkaline water, such as boiler water and boiler feed
water but is applicable also to waste waters when the interference is not excessive.
The amount of hydroxide ion may be a measure of the corrosiveness of the water toward those
materials subject to attack by basic medium.
There is no known method of chemical analysis by which the exact concentration of hydroxide
ion may be determined when other acid consuming components are present. Aluminates,
carbonates, chromates, phosphates, silicates, etc. and some organic matter affect the sample
titration to an undetermined degree, the amount being a function of both interference
concentration and sample pH. The effects of carbonates and phosphates, two components
commonly found in the water for which the method is normally used, are eliminated by the
addition of barium chloride in excess. No treatment is known for negating the other
interferences without destroying the hydroxide ion. However, if their combined concentrations
are very low as compared with that of the hydroxide ion (10% or less), the interference is
normally tolerable.
2. PRINCIPLE
The hydroxide ion is titrated with standard acid to the phenolphthalein end point after the
addition of barium chloride (BaC12) to precipitate dissolved carbonates and phosphates.
3. APPARATUS
3.1. Burette — 10 ml
3.2. Pipettes — 5 ml, 10 ml
3.3. Volumetric flasks — 100 ml, 500 ml, 1 litre
3.4. Conical flask — 250 ml
3.5. Weighing balance
3.6. Measuring cylinder — 100 ml.
4. REAGENTS
4.1. Barium chloride solution (10%)

Dissolve 10 g (BaCl2) Barium Chloride in distilled water and make up to 100 ml.
4.2. Sulphuric acid solution, standard
To be prepared from a BDH Convol (18032) or equivalent. Transfer quantitatively to
a 500 ml volumetric flask and make up to the mark with distilled water.
4.3. Phenolphthalein indicator
Dissolve 0.5 g of phenolphthalein in 50 ml of ethyl alcohol (95%) and dilute to 100
ml with distilled water.
GLAB-01(Revised) - Caustic Alkalinity Page 2 of 3

Page 3
5. PROCEDURE
Take 100 ml sample in a 250 ml conical flask. Add 3 drops of phenolphthalein indicator. Add 5
ml of barium chloride solution (10%) and shake. After 5 minutes, titrate with standard
sulphuric acid solution (0.01M). Note the titre reading when the pink colour of the solution
becomes colourless.
6. CALCULATIONS
0.01M Sulphuric acid = 0.02N Sulphuric acid
Caustic alkalinity as CaCO3, mg/l = Titre reading x 0.02 x 50 x 1000

Sample Volume, ml
= Titre reading x 0.02 x 50 x 1000
100
= Titre reading x 10
Where:
0.02 = Normality of H2SO4
50 = Equivalent weight of CaCO3
100 = Sample volume.
Caustic alkalinity as NaOH, mg/l = Titre reading x 0.02 x 40 x 1000

Sample volume, ml
= Titre reading x 0.02 x 40 x 1000
100
= Titre reading x 8
Where:
40 = Equivalent weight of NaOH
100 = Sample volume.
GLAB-01(Revised) - Caustic Alkalinity Page 3 of 3

Page 4
DUBAT ELECTRICITY AND WATER AUTHORITY
G-STATION LABORATORY METHODS
SECTION: WATER
THE DETERMINATION OF ALKALINITY IN WATER
(CARBONATE, BICARBONATE AND HYDROXIDE)
Method No : GLAB - 02
Page 5

ALKALINITY (CARBONATE, BICARBONATE AND HYDROXIDE)
G-LAB METHOD No.02
(By K.P. Abraham)
1. SCOPE
This test method covers the determination of alkalinity of water samples. By knowing the
phenolphthalein alkalinity (P-Alkalinity) and the methyl orange alkalinity (M-Alkalinity), the
carbonate, bicarbonate and hydroxide levels can be calculated.
2. PRINCIPLE
Alkalinity of water is it’s acid neutralizing capacity. It is the sum of all the filterable bases. It is
usually due to the bicarbonate, carbonate and hydroxide ions present in the sample, although
other ions such as phosphate, silicate and borates may partially contribute to the alkalinity.
The alkalinity is determined by titration with standard sulphuric acid solution using
phenolphthalein and then methyl orange in method - A and bromocresol green and methyl red
indicator in method - B, as indicators. The phenolphthalein alkalinity is measured at pH 8.3
whereas methyl orange alkalinity at pH 4.4.
3. APPARATUS
3.1 Automatic micro burette, 10 ml

3.2 Measuring cylinder, 100 ml
3.3 Indicator bottles
3.4 Conical flask, 250 ml
3.5 Volumetric flasks, 100 ml, 500 ml
3.6 Weighing balance
4. REAGENTS
4.1 Methyl orange indicator
Dissolve 0.04 g of methyl orange in 20 ml of alcohol. When dissolved add 80 ml of
distilled water and mix.
4.2 Phenolphthalein indicator
Dissolve 1 g of phenolphthalein is 60 ml of alcohol and then add 40 ml of distilled
water and mix.
4.3 Mixed Bromocresol green — Methyl red indicator
Dissolve 0.02 g of methyl red and 0.1 g of bromocresol green in 100 ml of ethyl
alcohol.
4.4 Sodium thiosulphate (N/20)
Dissolve 2.5 g of sodium thiosulphate (Na2S203.5H2O) in 100 ml
DM water and mix.
4.5 Standard acid solution (0.0lM H2SO4)
a 500 ml volumetric flask and make up to the mark with distilled water.
GLAB-02 (Revised) - Alkalinity Page 2 of 4

Page 6
5. PROCEDURE
Method - A
Measure 100 ml sample and transfer to the 250 ml conical flask. Add 4 drops of
phenolphthalein indicator and titrate against the 0.01M H2SO4, which is in the automatic
burette. Note the titre reading when the pink colour disappears. If there is no pink colour in the
beginning, note the reading as zero. Let the reading be Vp’,ml.. Then add 4 — 5 drops of
methyl orange indicator and continue the titration. Note the reading when the colour changes
from orange to red. Note this reading as VM’,ml.
Method — B
Measure 100 ml sample and transfer to the 250 ml conical flask, Add 4 drops of
phenolphthalein indicator and titrate against the 0.01M H2SO4, which is in the automatic
burette. Note the titre reading when the pink colour disappears. If there is no pink colour in the
beginning, note the reading as zero. Let the reading be ‘Vp’,ml. Then add 3 - 4 drops of N/20
sodium thiosulphate solution and 3 - 5 drops of bromocresol green - methyl red indicator and
continue the titration. Note the reading when the colour turns to purple gray. Note this reading
as VM ,ml.
6. CALCULATION
0.01M H2SO4 = 0.02N H2SO4
P - Alkalinity as mg CaCO3/l = Vp x 0.02 x 50 x 1000
Sample volume in ml
= Vp x 10
M - Alkalinity as mg CaCO3/l = VM x 0.02 x 50 x 1000
Sample volume in ml
= VM x 10
Where,
Vp = Titre reading, ml (Phenolphthalein end point)
VM = Titre reading, ml (Methyl orange end point)
100 ml = Sample volume
The results obtained from the phenolphthalein and methyl orange alkalinity determinations
offer a means for stoichiometric classification of the three principal forms of alkalinity present
in many water. The classification ascribes the entire alkalinity to bicarbonates, carbonates and
hydroxide and assumes the absence of other (weak) organic or inorganic acids, such as silicic,
phosphoric and boric acids. It further presupposes the incompatibility of hydroxide and
bicarbonate alkalinities. Because the calculations are made on a stoichiometric basis, ion
concentration in the strictest sense are not represented in the results, which may differ
significantly from actual concentrations especially at pH >10. According to this scheme
1. Carbonate (CO32-) alkalinity is present when phenolphthalein alkalinity is not zero but is
less than M-alkalinity.
2. Hydroxide (OH-) alkalinity is present if P-alkalinity is more than half the M-
alkalinity.
3. Bicarbonate (HCO3-) alkalinity is present if P-alkalinity is less than half the total
alkalinity.

Page 7
The relationship may be calculated by the following scheme, where ‘P’ is P-alkalinity and ‘M’
is M-alkalinity.
Select the smaller value of P or (M - P) . Then carbonate alkalinity equals twice the smaller
value. When the smaller value is P, the balance (M - 2P) is bicarbonate. When the smaller
value is (M-P), the balance (2P - M) is hydroxide. All results are expressed as CaCO3
ALKALINITY RELATIONSHIP
Result of Hydroxide Carbonate Bicarbonate

Titration concentration concentration concentration
as CaCO3 as CaCO3 as CaCO3
P=0 0 0 M
P < ½M 0 2P M—2P
P = ½M 0 2P 0
P>½ 2P-M 2(M—P) 0
P=M M 0 0
P = Phenolphthalein alkalinity, M = Methyl orange alkalinity.
For drinking water P < ½M.
Hence,
Carbonate as CaCO3 mg/l = 2P
Bicarbonate as CaCO3 mg/l = M-2P
Bicarbonate as HCO3 = (M—2P) x 61 = (M—2P) x 1.22
50
Where,
61 = Equivalent weight of HCO3
Reference APHA 2320 B

Page 8
DUBAT ELECTRICITY AND WATER AUTHORITY
SECTION: WATER
THE DETERMINATION OF AMMONIA (LOW RANGE)
Method No : GLAB - 03
Page 9

AMMONIA (DIRECT NESSLERIZATION METHOD)
G-LAB METHOD No.03
(By K.P. Abraham)
1. SCOPE
This method is suitable for the rapid routine determination of ammonia nitrogen in steam,
condensate and demineralised water.
2. PRINCIPLE
A sample aliquot is Nesslerized directly and the ammonia content determined

colourimetrically. The ammonia present in water forms reddish brown colour with Nessler’s
reagent, whose colour intensity is directly proportional to the ammonia content in water.
3. APPARATUS
3. 1. Drying Oven
3.3. Measuring cylinder, 50 ml
3.4. Pipette, 2 ml
3.5. Volumetric flask, 50 ml (10 Nos.), 1 lit, 200 ml.
3.6. Spectrophotometer, Jasco Model V 530
3.7. Beakers, 200 ml (10 Nos.)
3.8. Cell, 50 mm.
4. REAGENTS
4.1. Nessler’s reagent.
Readymade solution as purchased or this reagent can be prepared as follows:

Dissolve 100 g of anhydrous mercuric iodide (HgI2) and 70 g of anhydrous
potassium iodide (KI) in a small volume of water. Add this mixture slowly, with
stirring, to a cooled solution of 160 g of sodium hydroxide (NaOH) in 500 ml of
water. Dilute the mixture to 1 L. Store the solution in the dark for 5 days and filter
twice, either through a fritted glass crucible or glass fiber filter before using. If this
reagent is stored in a chemically resistant bottle out of reach of direct sunlight, it will
remain stable up to a period of 1 year.
4.2. Ammonia solution, Standard (1 ml = 1 mg NH3)

Dry ammonium chloride (NH4C1) at l00 °C overnight. Weigh 3.141 g of the dried
GLAB-03 (Revised) - Ammonia Page 2 of 3
Page 10
NH4C1 and dissolve in distilled water and make up to 11.

4.3. Ammonia solution standard (1 ml = 0.025 mg NH3)
Pipette 5.0 ml of the ammonia solution (1 ml = 1 mg NH3) to a 200 ml volumetric

flask arid make up to the mark.
5. CALIBRATION
Pipette 2.0, 4.0, 8.0, 12.0, 16.0 and 20.0 ml the standard ammonia solution (l ml = 0.025 mg
NH3) to 100 ml volumetric flasks. Make up to the mark with distilled water. This will give 0.5,
1.0, 2.0, 3.0, 4.0 and 5.0 mg/I Ammonia solutions. Transfer these to 200 ml beakers. Take 100
ml distilled water as blank in another beaker. Add 1 ml Nessler’s reagent to each including the
blank, Mix well and wait for 5 minutes for colour development. Measure the absorbance at 430
nm wavelength using 50 mm cells, against the distilled water blank. Plot a calibration graph
with absorbance against mg/l NH3. Calculate the factor ‘F’ which is equal to the mg/l NH3 per
unit absorbance.
6. PROCEDURE
Take 100 ml sample and 100 ml distilled water blank in 200 ml beakers. Add 1 ml Nessler’s
reagent to each. Mix well and wait for 5 minutes. Measure the absorbance using 50 mm cells at
a wavelength 430 nm. Note the absorbance. Concentration can be measured directly by
selecting File No.3 in JASCO, Model V 530 in G-Station laboratory.
7. CALCULATION
Ammonia, mg/l as NH3 = Absorbance x F

Where, F is the factor from the graph.
Note: a) Use only the clean supernatant portion of Nessler’s reagent.
b) Use only ammonia free distilled water for all solution preparation
and during determination.
c) As the intensity of colour fades with time, it is important to take

the reading just 5 minutes after adding Nessler’s reagent.
Reference: ASTM D 1426
GLAB-03 (Revised) - Ammonia Page 3 of 3

Page 11
SECTION: WATER
THE DETERMINATION OF AMMONIA

(SEA WATER & WASTE WATER)

Page 12

AMMONIA IN SEA WATER & WASTE WATER
G-LAB METHOD No.04
(By K.P. Abraham)
1. SCOPE
This method is suitable for the routine determination of ammonia nitrogen in seawater and
wastewater.
2. PRINCIPLE
Turbidity, colours, and substances precipitated by hydroxyl ion such as magnesium and
calcium, interfered is removed by preliminary distillation. Sample is buffered at pH 9.5 with a
borate buffer to decrease hydrolysis of cyanates and organic nitrogen compounds. It is distilled
into a solution of boric acid.
An aliquot of the distillate is Nesslerized directly and the ammonia content determined
colourimetrically. The ammonia present in water forms reddish brown colour with Nessler’s
reagent, whose colour intensity is directly proportional to the ammonia content in water.
3. APPARATUS
3.1. Drying Oven
3.4. Pipette, 2 ml
3.5. Volumetric flask, 100 ml (10 Nos.), 1 lit, 200 ml
3.6. Spectrophotometer, JASCO Model V 530
3.7 Beakers, 200 ml (10 Nos.)
3.8 Cell, 50 mm.
3.9. Distillation apparatus consisting of 500 ml round bottom flask, spray trap, bend tube,
condenser and receiver, heater etc.
4. REAGENTS
4.1. Nessler’s reagent.
Readymade solution as purchased or this reagent can lie prepared as follows:

Dissolve 100 g of anhydrous mercuric iodide (HgI2 ) and 70 g of anhydrous
potassium iodide(KI) in a small volume of water. Add this mixture slowly, with
stirring, to a cooled solution of 160 g of sodium hydroxide (NaOH) in 500 ml of
water. Dilute the mixture to 1 L. Store the solution in the dark for 5 days and filter
twice, either through a fritted glass crucible or glass fiber filter before using. If this
GLAB-04 (Revised) - Ammonia in Seawater& Waste water Page 2 of 4

Page 13
reagent is stored in a chemically resistant bottle out of reach of direct sunlight, it will
remain stable up to a period of 1 rear,
4.2. Ammonia solution, Standard (1 ml = 1 mg NH3)
Dry ammonium chloride (NH4C1) at l00 °C overnight. Weight 3.141 g of the dried
NH4C1 and dissolve in distilled water and make up to 11itre.
4.3. Ammonia solution standard (1 ml = 0.025 mg NH3)
Pipette 5.0 ml of the ammonia solution (1 ml = I mg NH3) to a 200 ml volumetric

flask and make up to the mark.
4.4. Borate buffer
Add 88 ml of 0.1M NaOH solution to 500 ml approximately 0.025M sodium

tetraborate (Na2B4O7) solution (9.5 g Na2B407.H2O/L) and
dilute to 1L.
4.5. Sodium hydroxide (1 M)
Dissolve 40 g NaOH in water and dilute to l L.
4.6. Dechlorinating agent.
Dissolve 3.5 g of sodium thiosulphate (Na2S2O3.5H20) in water and dilute to 1L.
4.7. Boric acid (2%)
Dissolve 20 g boric acid (H3BO3) in water and dilute to 1L with ammonia free water.
4.8. DPD Tablet No.1
5. CALIBRATION
Pipette 2.0, 4.0, 8.0, 12.0, 16,0 and 20.0 ml the standard ammonia solution (l ml = 0.25 mg
NH3) to 100 ml volumetric flasks. Make up to the mark with distilled water. This will give 0.5,
1.0, 2.0, 3.0, 4.0 and 5.0 mg/l Ammonia solutions. Transfer these to 200 ml beakers. Take 100
ml distilled water as blank in another beaker. Add 1 ml Nessler’s reagent to each including the
blank. Mix well and wait for 5 minutes for colour development. Measure the absorbance at 430
nm wavelength using 50 mm cells, against the distilled water blank. Plot a calibration graph
with absorbance against mg/l NH3. Calculate the factor ‘F’ which is equal to the mg/l NH3 per
unit absorbance.
6. PROCEDURE
A. Distillation
Free chlorine in the sample is checked using DPD No.1 tablet. If chlorine is present,
eliminate it by adding 0.5 ml of dechlorinating agent to remove 1 mg/L residual chlorine in
250 ml sample. Take 250 ml chlorine free sample in a beaker and add 15 ml borate buffer
and mix. Check the pH and adjust the pH to 9.5 using sodium hydroxide( 1M) solution.
Transfer it quantitatively to the distillation flask and connect the condenser unit. Take 25
ml boric acid solution (2 %) in the receiving beaker and keep the tip of the condenser
dipped in the solution, Start the distillation at a rate of 6 ml/min. Collect 200 ml distillate

Page 14
and transfer that to a 250 ml volumetric flask and make up that to 250 ml with ammonia
free DM water. Take 250 ml ammonia free DM water and do the distillation, as the
sample, for the reagent blank.
B. Ammonia determination
Take 100 ml sample distillate and 100 ml blank distillate solutions in 200 ml beakers. Add
2 ml Nessler’s reagent (an excess that raises the pH to the desired level) to each. Mix well
and wait for 5 minutes. Measure the absorbance using 50 mm cells at a wavelength 430
nm. Note the absorbance. Concentration can be measured directly by selecting file No.3 in
JASCO, Model V 530 in G - Station laboratory.
7. CALCULATION
Ammonia, mg/l as NH3 = Absorbance x F

Note: a) Use only the clean supernatant portion of Nessler’s reagent.
b) Use only ammonia free distilled water for all solution preparation
and during determination.
c) As the intensity of colour fades with time, it is important to take
the reading just 5 minutes after adding Nessler’s reagent.

APHA 4500-NH3 (B)

Page 15
DUBAI ELECTRICITY AND WATER AUTORITY
LABORATORY METHOD
SECTION: WATER
(THE ATOMIC ABSORPTION/EMISSION SPECTROPHOTOMETRY)

Date issued : 2nd November 1996
Page 16

THE ATOMIC ABSORPTION / EMISSION SPECTROPHOTOMETRY.
G-LAB METHOD NO.05
(By K.P. Abraham)
1. SCOPE
This method covers the determination of calcium, copper, iron, potassium, magnesium,
sodium, nickel, lead, titanium, vanadium and zinc in water samples. Samples, which can be
analysed includes cooling water, evaporator acid cleaning samples, drinking water, effluent
water, steam samples etc.
2. PRINCIPLE
Atomic absorption spectrophotometry make use of the fact that neutral or ground state atoms of
an element can absorb electromagnetic radiation over a series of very narrow, sharply defined
wavelengths. The sample, in solution, is aspirated as a fine mist into a flame where it is
converted to an atomic vapour. Most of the atoms remain in the ground state and are therefore
capable of absorbing radiation of a suitable wavelength. This discreet radiation is usually
supplied by a hollow cathode lamp which is a sharp line source consisting of a cathode
containing the element to be determined along with a tungsten anode.
When a sufficient voltage is impressed across the electrodes, the filter gas is ionised and the
ions are accelerated towards the cathode. As these ions bombard the cathode they cause the
cathode material to “sputter” and form an atomic vapour in which atoms exist in an excited
electronic state. In returning to the ground state the lines characteristic of the element are
emitted and pass through the flame where they may be absorbed by the atomic vapour. Since,
only the test element can absorb this radiation, the method becomes very specific in addition to
being sensitive.
In atomic emission spectroscopy, the metal is excited from the energy imparted to it thermally
by the flame and then as it returns to the ground state it emits radiation at a characteristic
wavelength. This radiation is then isolated by a monochromator and subsequently its intensity
is directly proportional to the concentration of the element present.
3. APPARATUS
3.1. Atomic absorption / emission spectrophotometer

Perkin-Elmer Model 3100
3.2. Funnel
3.3. Whatman filter paper No.41
3.4. Pipettes, 1, 2, 4, 5, 10 & 20 ml.
3.5. Volumetric flasks, 1 litre, 100 ml (15 Nos.)
GLAB-05 (Revised) - Atomic Absoption & Emission Spectrometry Page 2 of 7

Page 17
4. REAGENTS
4.1. Sulphuric acid (1M)

Add carefully 56 ml concentrated sulphuric acid to 500 ml distilled water and dilute to
1 litre.
4.2. BDH standard solutions (1ml = 1mg)

For calcium, copper, Iron, potassium, magnesium, sodium, nickel, lead, titanium,
vanadium and zinc.
4.3. Standard Solutions (1ml = 0.01mg)

Pipette 10.0ml each of the above standards except titanium and vanadium to 1000ml
volumetric flasks and make up to the marks with DM water.
4.4 Standard Solutions (1ml = 0.1mg)

Pipette.10.0 ml of titanium and vanadium standards (1 ml = 1 mg) to 100 ml
volumetric flasks and make up to the mark with DM water.
4.5. Working standard solutions

Pipette Ca, Cu, Fe, K, Mg, Na, Ni, Pb & Zn standard solutions (1 ml = 0.01 mg) and
Ti & V standard solution (1 ml = 1 mg) as described below in to 100 ml volumetric
flasks. Make up the volume with distilled water. Store the solution in polythene
bottles.
ml of standard solution from Concentration

Working (4.3)/(4.4) for (mg/l)
standards No Ca,Cu,Fe,K,Mg,Na,Ni,Pb,Ti,V
and Zn Ti & V Rest all
1 1 1.0 0.1
2 2 2.0 0.2
3 4 4.0 0.4
4 6 6.0 0.6
5 8 8.0 0.8
6 10 10.0 1.0
7 20 20.0 2.0
8 30 30.0 3.0
9 40 40.0 4.0
10 50 50.0 5.0
11 60 60.0 6.0
12 70 70.0 7.0
13 80 80.0 8.0
14 90 90.0 9.0
15 100 100 10.0
Page 18
5. PROCEDURE
5.1 Take 100 ml of the sample. Add 5 ml of 1M sulphuric acid and boil for 15 minutes. Cool
and make upto 100 ml with distilled water. Filter the sample, if it is turbid, using a
whatman No.41 filter paper.
5.2. Put ON the ventilation.

Open the acetylene and nitrous oxide (if needed) gas cylinders and note the bottle
pressure.
Fix the required burner head. (Air-Acetylene — 10 cm (Wide),

N2O - Acetylene — 5 cm (Narrow)).
Fix the required Hollow cathode lamp. (For atomic absorption mode)
Set the wavelength and the slit width as specified in the Standard conditions. (Table 2 for
atomic absorption, Table 3 for atomic emission) and the slit height to HIGH position.
Switch ON the instrument.
Set the instrument parameters through the parameter entry mode.
Press ENERGY.
Maximize the energy by adjusting the lamp position and adjusting the wavelength.
Set the normal operating conditions, which is specified in Table-1.
Ignite the flame.
When using the atomic emission mode, by keeping the highest standard press GAIN to
make the automatic gain control adjustment.
Maximize the energy by adjusting the burner height and angle with the aspiration of the
highest standard.
Activate the printer, if printing is required.
Enter the data collection mode by pressing DATA.
Run the blank and press A/Z.
Keep the first standard to aspirate, enter ‘0’ and press ‘Calib.’
Keep the standards to aspirate in sequence and press ‘Calib.’ each time
Page 19
After completing the calibration keep the blank and press READ to see the blank reading.
Blank can be auto zeroed any time by aspirating the blank and pressing A/Z.
Reslope can be done any time, if a reslope standard is entered in the parameter entry, by
keeping the reslope standard for aspiration and pressing RESLOPE.
Any standard can be rechecked and entered in the graph by keeping the standard to
aspirate and enter the standard number and press ‘Calib.’
Analyse the sample by keeping the sample to aspirate and press READ.
A continuous absorption reading can be noted by pressing CONT. button.
After completing the samples, aspirate DM water to clean the burner head.
Turn the gas control switch on the pneumatic control panel to OFF.
Close the acetylene / nitrous oxide gas source main valves.
Turn the gas control switch to air or N2O (if used) to allow them to bleed. The red
interlock light on the ignite button should turn ON.
Turn the gas control switch to OFF.
Close the air supply main valve.
Turn OFF the Hollow cathode lamp (if used) by entering the lamp current as ‘0’.
(Parameter # 1).
Turn the instrument OFF.
Turn the vent OFF.
Close the gas cylinders.

Page 20
Normal Operating Conditions
Table - 1
S1.No. Parameters Air—Acetylene N2O—Acetylene

1 Air supply pressure - Bar 4.0 4.0
2 Syn. Air cylinder pressure - Bar >15 >15
3 Acetylene pressure - Bar 0.9 1.0
4 Acetylene cylinder pressure - Bar >5.2 >5.2
5 Nitrous oxide pressure - Bar - 4.0
6 Nitrous oxide cylind. pressure - Bar - >10
7 Oxidant flow - Units 4.0 4.0
8 Fuel flow - Units 2.5 3.5
9 Flame condition Lean, Blue Redzone,1”
10 Sample flow rate (aqueous) - ml/min 8 8
11 Sample flow rate (solvent) - ml/min 3 3
Standard Conditions
Table - 2 ( Atomic absorption)
Wave
Flame Slit L.Cur Sen.C L.Rng
No Element length
(Gases) (nm) (mA) Mg/l Mg/l Remarks
(nm)
1 Calcium - Ca Air-Acety 422.7 0.7 20 4.0 5.0
2 Copper - Cu Air-Acety 324.8 0.2 30 4.0 5.0
3 Iron - Fe Air-Acety 248.3 0.2 30 5.0 5.0
4 Potassium - K Air-Acety 766.5 1.4 12 2.0 2.0 0.1%alk.add
5 Magnesium- Mg Air-Acety 285.2 0.7 20 0.3 0.5
6 Sodium - Na Air-Acety 589.0 0.4 12 0.5 1.0 0.1%alk.add
7 Nickel - Ni Air-Acety 231.1 0.2 30 10.0 5.0
8 Lead - Pb Air-Acety 283.3 0.7 10 20.0 20.0
9 Titanium - Ti N2O-Acety 364.3 0.2 40 80.0 100
10 Vanadium - V N2O-Acety 318.4 0.7 40 90.0 200
11 Zinc - Zn Air-Acety 213.9 0.7 20 1.0 1.0

Page 21
Table - 3 ( Atomic emission)
Wavelength
No Elements Flame(Gases) Slit(nm)
(nm)
1 Calcium - Ca N2O—Acetylene 422.7 0.2
2 Copper - Cu N2O—Acetylene 324.8 0.2
3 Iron - Fe N2O—Acetylene 372.0 0.2
4 Potassium - K Air—Acetylene 766.5 0.4
5 Magnesium –Mg N2O—Acetylene 285.2 0.7
6 Sodium - Na Air—Acetylene 589.0 0.2
7 Nickel - Ni N2O—Acetylene 341.5 0.2
8 Lead - Pb N2O—Acetylene 405.8 0.2
9 Titanium - Ti N2O—Acetylene 399.9 0.2
10 Vanadium - V N2O—Acetylene 437.9 0.2
11 Zinc - Zn N2O—Acetylene 213.9 0.2
6. CALCULATION
6.1. Read off the concentration values for the samples from the instrument display as
mg/litre.
6.2. Take into account, the dilution factor, if any.
7. NOTE
7.1. Follow, all the safety instructions which is recommended for the instrument.
7.2. If the sample concentration exceeds the standard concentration give necessary dilution
to bring the concentration in the limit of standard concentration.
Reference: Operation manual for Perkin-Elmer Model 3100.

Page 22
SECTION: WATER
THE DETERMINATION OF BIOLOGICAL OXYGEN DEMAND
(BOD TRACK METHOD)
Method No. : GLAB-06

Date issued : 24th April 2006
Prepared by : Abraham & Varkey
Page 23

BIOLOGICAL OXYGEN DEMAND (BODTrack METHOD)
G-LAB METHOD No.06
(By K.P. Abraham & Philip Varkey)
1. SCOPE
This method is used for the determination of Biological Oxygen Demand (BOD) of
wastewater, effluents and polluted waters. BOD analysis is a key test in water
pollution and water treatment process control.
2. PRINCIPLE
Bacteria in the sample uses, oxygen to oxidize organic matter within the sample
bottles. The air in the bottle above the sample contains 21% oxygen and replenishes
the oxygen used by the bacteria. During the test period, stir bars continually rotate
with in each bottle. The carbon dioxide must be removed from the system so the
measured pressure difference is proportional only to the amount of oxygen used.
Stirring helps transfer oxygen from the air to the sample and helps simulate natural
conditions. BOD track is sealed to prevent external atmospheric pressure changes
from affecting the BOD readings. Pressure sensors monitor air pressure within the
sample bottles, and when the air pressure drops, the pressure change is converted to
mg/ l BOD. The carbon dioxide is removed from the system by the lithium hydroxide
crystal placed in the seal cup of each sample bottle before testing.
3. APPARATUS
3.1 Incubator
3.2 BOD Track
3.3 BOD amber bottle
3.4 Seal cup & Magnetic stir bar
3.5 Stopcock grease
4. REAGENT
4.1 Lithium Hydroxide power pillows
4.2 BOD Nutrient buffer pillows
4.3 BOD Seed ( Polyseed Inoculum )
GLAB-06 (Revised) - B.O.D. (HACH method) Page 2 of 5

Page 24
5. PROCEDURE
5.1 Seed preparation
Take 3 Liter of distilled water add one nutrient buffer pillows to the water.
Aerate the water. Take 500 ml of water add one polyseed capsule into dilution
water to rehydrate (Hach cat 24712-00 ) .Aerate and stir for one hour. Allow
to settle for one hour.
5.2 Sample analysis
Heat or cool the sample to within 2°C of its incubation temperature (typically
20 °C)
Selection of sample volume
BOD Range ( mg/l) Required volume ( ml)
0- 35 420
0-70 355
0-350 160
0-700 95
5.2.1. Using a clean graduate cylinder add 417 ml of sample and 3 ml of BOD
seed into the BOD Track sample bottles.
5.2.2. Place a magnetic stir bar in each sample bottle. Add the content of one
BOD nutrient buffer pillows to each bottle for optimum bacteria growth.
If simulation of original sample characteristic is required do not add
nutrient buffer.
5.2.3. Apply stopcock grease to the seal lip of each bottle and to the to each
seal cup. Place seal cup in the neck of each bottle
5.2.4. Using a funnel add the content of one lithium hydroxide to each seal
cup. Do not allow lithium hydroxide particle to fall into the sample, if
this occurs discard the sample and prepare a fresh one.
5.2.5. Place the bottles on the chassis of the BOD track. Connect the
appropriate tube to the sample bottle and firmly tighten the cap. Each
tube is tagged with the channel number, and the channel number setup
will be reflected on the control panel.
Page 25
5.2.6. Switch on the incubator and adjust the temperature to 20 °C. Place the
BOD Track in the incubator. The Place the seal cup in the neck of each
bottle.Connect the electrical plug and turn the instrument on. Make sure
all stir bars are rotating properly.
5.2.7. To start the test press the channel number corresponding in the bottle.
Each channel (1-6 ) must be started individually.
5.2.8. Press and hold the ON key .A menu for selecting the BOD range will be
displayed. For 0-35 mg/l < (left) key. For 0-70 mg/l press the < key a
second time. Press and hold the ON key to start a test. An empty graph
for the selected channel is displayed. The status annunciate in the lower
right of the display will show DLY for the first hour of the test, during
which no data are taken. This allows the instrument, bottles, and sample
to equilibrate to the incubation temperature. After the first hour the
annunciate shows RUN, indicating that the channel is actively collecting
data. The BOD test automatically ends after the selected test duration of
5.25 days. The status annunciator in the lower right of the display will
change from RUN to END.
6. CALCULATION
BOD mg/l = BOD –BOD Seed correction
BOD Track Calibration check

(Providing Manometric BOD test and apparatus accuracy)
1. Nutrient buffer solution
Take 3 Liter of distilled water add one nutrient buffer pillows to the water.
Aerate the water.
2. Seed preparation
Take 500 ml of nutrient buffer solution add one polyseed capsule (Hach cat
24712-00 ). Aerate by stir for one hour. Allow to settle for one hour.(Best
results are obtained by drawing a aliquot one or two inches below surface. Be
careful not to draw from the bottom, as the presence of undisssolved material
may lead to non-uniform results)

Page 26
3. Take 6 BOD amber bottles.

3.1.Bottle # 1 420 ml of Nutrient buffer solution 420 ml (BOD rang 0-35
mg/l)
3.2.Bottle # 2 417 ml of Nutrient buffer solution and 3 ml of seed.(BOD
Range 0-35 mg/l )
3.3.Bottle # 3 Take 415 ml of Nutrient buffer solution , 3.0 ml of seed ,2 ml of
3000 ppm BOD Standard.( BOD range 0-35 mg/l)- 14.3 mg/l Std.
3.4. Bottle # 4 155ml of Pure seed water ( BOD Range 0-350 mg/l)- SEED
BOD
3.5.Bottle # 5 133 ml of Nutrient buffer solution , 15 ml of seed and 7 ml of
BOD standard ( 3000mg/l). (BOD range 0-350 mg/l)- Observed BOD
3.6.Bottle # 6 415 ml of Nutrient buffer solution , 3.0 ml of seed ,2 ml of
3000 ppm BOD Standard.( BOD range 0-35 mg/l)- 14.3 mg/l Std.
Check for BOD using sample analysis procedure starting from step 5.2.2
CALCULATION
BOD of sample = (BOD Observed) –(Decimal fraction of seed used x BOD Seed)
(Decimal fraction of sample used )
Example : A seeded sample is 10 % seed and 90 % sample (by volume )
The observed BOD is 60 mg/l and the pure seed BOD is 150 mg/l
BOD Sample = (60 mg/l) – (0.10x150 mg/l) = 50 mg/l
0.90
BOD for 14.3 mg/l standard = ( BOD mg/l ) – (0.007x BOD Seed )
0.993
BOD for 3ml seed correction = BOD of orginal x Vol of seed

Vol. Of sample
Note: BOD Test for all the BOD track to be carried out at the same as by taking 415
ml of Nutrient buffer solution , 3.0 ml of seed, 2 ml of 3000 ppm BOD
Standard.( BOD range 0-35 mg/l) - 14.3 mg/l Std.
Reference: HACH BODTrack Instruction manual

Page 27
SECTION: WATER
THE DETERMINATION OF BIOLOGICAL OXYGEN DEMAND
(WINKLER OR IODOMETRIC METHOD)

Date issued : 28th March 1995
Page 28
TEST METHOD FOR EFFLUENT WATER

BIOLOGICAL OXYGEN DEMAND BY WINKLER OR IODOMETRIC METHOD
G-LAB METHOD No.07
(By K.P. Abraham)
1. SCOPE
This method is used for the determination of Biological Oxygen Demand (BOD) of
wastewaters, effluents and polluted waters. BOD analysis is a key test in water pollution and
waste treatment process control.
2. PRINCIPLE
The biological oxygen demand (BOD) determination is an empirical test in which standardized
laboratory procedures are used to determine the relative oxygen requirements. The test
measures the oxygen required for the biological degradation of organic material (carbonaceous
demand) and the oxygen used to oxidize inorganic material such as sulfides and ferrous ion. It
also may measure oxygen used to oxidize reduced forms of nitrogen, (nitrogenous demand)
unless, their oxidation is prevented by an inhibitor.
The dilution method involves the treatment of the sample with aerated water and incubation for
5 days at 20°C. The dissolved oxygen content of the water before and after the incubation
period is determined by the azide method, the difference giving the BOD of the sample after
allowance has been made for the dilution of the sample.
3. APPARATUS
3.1. Narrow mouthed glass-stoppered bottles of a nominal capacity 250 ml. Iodine flasks
with flared necks and ground glass stoppers are ideal for this purpose, having a total
volume of 300 ml when stoppered. New bottles should be cleaned with either SN
hydrochloric or sulphuric acid and thoroughly rinsed with water.
3.2. Incubator maintained at 20 °C (± 0.5°C) from which light is excluded to prevent

algae in the sample from producing oxygen by photosynthetic action.
3.3. Burette, 25 ml.
3.4. Measuring cylinder.
3.5. Pipettes.
3.6. Volumetric flasks, 100 ml, 200 ml and 1000 ml.
4. REAGENTS
4.1. Water
Water used for solution preparation and dilution must be of the highest quality and
must be free of copper (<0.01 mg/litre), chlorine, chloramines, caustic alkalinity,
organic material or acids. Water from the laboratory still, which has been passed
through the Millipore system, is suitable.
GLAB-07 (Revised) - B.O.D. (Winkler method) Page 2 of 6

Page 29
4.2. Ferric chloride solution

Dissolve 0.250 g of ferric chloride (FeCl3.6 H20) in water and make up to 1 litre in a
volumetric flask. Store in a glass stoppered bottle.
4.3. Calcium chloride solution

Dissolve 27.5 g of calcium chloride (CaC12) or equivalent if hydrated calcium
chloride is used in water and make up to 1 litre in a volumetric flask. Store in a
glass stoppered bottle.
4.4. Magnesium sulphate solution
Dissolve 22.5 g of magnesium sulphate (MgS04.7H20) in water and make up to 1 litre
in a volumetric flask. Store in a glass stoppered bottle
4.5. Phosphate buffer solution

Dissolve 8.5 g of potassium dihydrogen phosphate (KH2PO4),2l.75 g
dipotassium hydrogen phosphate (K2HPO4), 33.4 g disodium hydrogen
phosphate (Na2HPO4.7H2O), and 1.7 g ammonium chloride (NH4C1) in about
500 ml water and dilute to 1 L. This should give a solution of pH 7.2 which
should be checked.
Note Any of these solutions showing signs of precipitates or growths should be
discarded.
4.6. Manganous sulphate solution (48% W/V)

Dissolve 480 g of manganese sulphate (MnSO4.4H2O) in distilled water and make up
to 1 L. No iodine should be liberated when 1 ml of the reagent is added to 50 ml of
acidified potassium iodide solution.
4.7. Sulphuric acid

Concentrated sulphuric acid (H2SO4 Sp.gr 1.84).
4.8. Alkaline potassium iodide solution

Dissolve 500 g of sodium hydroxide (NaOH) (or700 g KOH) and 135 g of sodium
iodide (NaI) (or 150 g KI) in distilled water and dilute to 1 L. Add 10 g of sodium
azide (NaN3) dissolved in 40 ml distilled water. Potassium and sodium salts may be
used interchangeably.
4.9. Orthophosphoric acid (85 — 90%)

Use the Analar grade reagent (S.G. 1.75 approx. 88%).
4.10. Sodium thiosulphate solution, standard (0.01 M)

Dissolve 2.482 g ± 0.001 g of sodium thiosulphate (Na2S2O3 5H2O) in distilled water
and make upto 1L.
4.11. Standard Iodine solution (0.05 M)

To be prepared from BDH Convol, Prod. No. 18038 or equivalent. Transfer
quantitatively to a 1 litre volumetric flask and make up to the mark with
distilled water.
4.12. Starch indicator

0.5 g of soluble starch is made to a paste with little cold water and then add 25 ml of
boiling water. Boil until a clean solution is obtained.

Page 30
5. STANDARIDIZATION
Pipette 5.0 ml of the standard iodine solution (0.05 M) to 250 ml Erlenmeyer flask. Take the
prepared sodium thiosulphate solution in a 100 ml burette. Start titration with constant shaking.
When the iodine colour is faint add 2 ml of starch solution and continue the titration. The end
point is noticed by the disappearance of the blue colour. Note the titre reading.
0.05 M Iodine = 0.1 N Iodine
Normality of Sodium thiosulphate = 0.1_X_5____

Titre reading, ml
If the normality obtained differs from 0.01 N, adjust the normality to 0.01 N by adding
calculated amount of Na2S2O3.5H2O or by diluting with distilled water. Standardize again. This
will provide a 0.01 M sodium thiosulphate solution.
6. PROCEDURE
6.1. The test should be carried out as soon as possible after the sample has been taken. If
samples are kept at room temperature for several hours a very appreciable change
may occur in the BOD depending on the character of the sample (i.e. 40% decrease
over eight hours). If samples cannot be dealt with at once they should be stored at 50C
in a refrigerator until required.
6.2. Prepare the dilution water as follows:
Add 1 ml of ferric chloride solution, calcium chloride solution, magnesium sulphate

solution and phosphate buffer to each litre of distilled water contained in a suitable
beaker. Place the beaker in the water bath and allow it to attain incubation
temperature (20°C ± 0.5°C). Using compressed air or oxygen, saturate the volume of
water with oxygen by vigorous bubbling of the gas through the liquid (15 mints). Use
this water as soon as possible after preparation.
6.3. If the pH of the sample is not in the range 6.5 - 8.5 add sufficient alkali or acid to
bring it within the range. Determine the amount of acid or alkali to be added on a
separate sample.
6.4. Unless the BOD of the sample is already known the required degree of dilution will
not be known and more than one dilution will have to be made. The following gives
approximate dilutions for one volume of sample:
Purified sewage effluent - 1 - 9 volumes dilution water

Settled sewage - 14 - 99 volumes dilution water
Crude sewage - 29 - 199 volumes dilution water
Dilute a suitable volume of the sample in a volumetric flask (1litre) with the freshly
aerated water. Samples, which are been stored in a refrigerator, should be allowed to
reach room temperature before dilutions are made. Mix the solution by careful
repeated inversion of the flask. Violent agitation must be avoided.
6.5. Transfer the diluted solution to two iodine flasks by careful pouring till the water level
is above the ground glass necks. Allow the bottles to stand for a few minutes and then
tap gently to remove any small air bubbles. Insert the stopper carefully ensuring that
no air bubbles are trapped underneath.

Page 31
6.6. Repeat the above step using the dilution water only.
6.7. Place one bottle of the sample together with one of the dilution water in the
incubator bath at 20°C (+ 0.5°C) and incubate in the dark for five days (± 3 hours).
6.8. Determine the initial concentration of dissolved oxygen in the other iodine flask
containing the diluted sample and also on the dilution water blank using the azide
method.
6.9. Azide method for the determination of dissolved oxygen
6.9.1. To the sample in the iodine flask, pipette 2 ml manganous sulphate

solution followed by 2 ml alkaline iodide- azide solution. The tip of the
delivery pipette in both cases should be inserted well below the surface of
the liquid.
6.9.2. Replace the stopper carefully so as to avoid inclusion of air bubbles and
thoroughly mix the contents by vigorously
inverting and rotating the bottle several times.
When the precipitate has settled to the lower third of the bottle, repeat the
mixing and then allow the precipitate to settle completely leaving a clear
supernatant liquid.
6.9.3. Add 2 ml sulphuric acid (Concentrated) or 85 — 95% phosphoric acid (if

the presence of ferric salts is suspected) by pipette at the surface of the
liquid. Replace the stopper and thoroughly mix the contents by rotation.
6.9.4. Measure a suitable volume V ml of the sample into a 250 ml conical flask
and titrate the yellow colour with 0.0l M sodium thiosulphate to pale yellow
colour. Add by pipette 2 ml of the starch solution and continue the titration
till the colour changes from blue to colourless. Let the titre = x ml.
6.10. Repeat stages of 6.9 with the aerated dilution water.

Let the titre = a ml.
6.11. After five days (± 3 hours) remove the iodine flasks from the
incubation and repeat stages of 6.9 and 6.10 for the sample and the aerated dilution
water.
Let titre for sample = y ml
Let titre for dilution water = b ml
7. CALCULATION
BOD5, mg/L = [(x-y) - (a-b) n ] x 0.01 (n+l) 1000 x 8

n+l V
Where
x = Volume 0.01M thiosulphate required for Vml of original diluted sample(ml)
y = Volume 0.01)1 thiosulphate required for Vml of incubate diluted sample(ml)
a = Volume 0.01M thiosulphate required for Vml of original aerated dilution

water(ml)
b = Volume 0.0lM thiosulphate required for Vml incubated dilution water (ml)

Page 32
V = Volume of sample taken for titration (ml)
n = Volume of dilution water to 1 volume of sample (ml)
n/n+l = Correction factor for volume occupied by sample compared to volume

of dilution water used
0.01 = Normality of sodium thiosulphate
8 = Equivalent weight of oxygen
Notes: It may be desired from time to time to check the precision and bias of
the method using a standard sample. For this purpose the following
procedure should be used
Dissolve 0.150 g (± 0.001 g) each of glucose and glutamic acid (both dried at 108°C for one
hour) in a litre of water. This solution should be freshly prepared when required.
Make up a 1 in 50 dilution using the seeded aerated dilution water and determine the BOD in
the usual way.
The BOD should be 220 mg/litre (± 20 mg/litre).
If the result is outside these limits then some defect in the reagents or experimental technique
should be suspected.
Reference: APHA 5210 B

Page 33
DUBAI ELECTRICITY AND WATER. AUTHORITY
SECTION: WATER
THE DETERMINATION OF FREE CARBON DIOXIDE

Page 34

FREE CARBON DIOXIDE
G-LAB METHOD No.08
(By K.P. Abraham)
1. SCOPE
This method is suitable for estimating free carbon dioxide ( CO2 ) content of water for routine
control purpose, as in water treatment systems
2. PRINCIPLE
Free CO2 is reacted with sodium carbonate (Na2 CO3 ) to form sodium bicarbonate (NaHCO3 ) .
The end point of the reaction is detected by means of a pH colour indicator, phenolphthalein.
3. APPARATUS
3. 1. Glass cylinder with stopper, 100 ml
3.2. Automatic burette, 10 ml
3.3. Volumetric flasks, 500 ml, 1 litre
3.4 Indicator bottle
3.5. Pipette, 100 ml
4. REAGENTS
4 .1. Sodium carbonate, standard (0 . 01M)

To he prepared from an ampoule, BDH Convol, Product No.18021 or equivalent.
Transfer quantitatively to a 500 ml volumetric flask and make up to the mark . Shake
well and pipette 100 ml from this to another 500 ml volumetric flask and make up to
the mark with distilled water.
OR
Prepare sufficient CO2 free water by boiling distilled water for 20 minutes and
cooling rapidly before use. Dry about 2 g of sodium carbonate anhydrous at 300 °C
for 2 hours and allow to cool in a desiccator. Dissolve 1.06 g of the dried sodium
carbonate in about 100 ml of CO2 free distilled water and make up to exactly 1 litre in
a volumetric flask with CO2 - free water.
Dissolve 1.0 g of phenolphthalein in 100 ml of 95% ethyl alcohol
5. PROCEDURE
Collect the sample directly to a 100 ml standard measuring cylinder with stopper. Fill
completely and keep it stoppered. Keep the sample, until tested, at a temperature lower than
that at which the water was collected, Carry out the analysis as soon as possible to minimize
the loss.
GLAB-08 (Revised) - Carbon dioxide, Free Page 2 of 3

Page 35
Take 100 ml sample in the measuring cylinder, in which the sample is collected, by throwing
the excess sample out. Add 3 drops of phenolphthalein indicator. Close the cylinder and shake
gently. Titrate with the standard sodium carbonate (0.01 M). The end point is noted by the
appearance of a pink colour in the sample when looked from the top of the cylinder against a
white background. Note the titre reading,
6. CALCULATION
Free CO2 = Titre reading x 0.01 x 44 x 1000 = Titre reading X 4.4

100

APHA 4500 CO2 (C).
GLAB-08 (Revised) - Carbon dioxide, Free Page 3 of 3

Page 36
SECTION WATER
THE DETERMINATION OF CHEMICAL OXYGEN DEMAND
(COD REACTOR METHOD)

Page 37

CHEMICAL OXYGEN DEMAND
[COD Reactor METHOD]
G-LAB METHOD No.09
(By K.P. Abraham & Philip Varkey)
1. SCOPE
This method covers the determination of the quantity of oxygen that certain impurities in water
will consume, based on the reduction of a dichromate solution under specified conditions. This
method can be applied to cooling waters, industrial effluents, domestic sewage etc. It can
measure COD values in the range of 50-800 mg/l. The measurement of COD may be used for
evaluating the organic contaminants and for evaluating the treatment and control of industrial
wastewater.
2. PRINCIPLE
Most organic and oxidisable inorganic substances present in water are oxidized by a standard
potassium dichromate solution in 50 % sulphuric acid solution. The oxidation of many
organics is facilitated by the use of silver sulphate, which acts as a catalyst in the reaction. The
oxidation of up to 1000 mg/l of chloride ion is inhibited by the addition of mercuric sulphate to
form a stable and soluble mercuric chloride complex.
3. APPARATUS
3.1. COD Reactor
3.2. Spectrophotometer – HACH 2010
3.3. Pipette
3.4. Volumetric flask
4. REAGENTS
4.1. COD Digestion Reagent 0 to 150 ppm ( Low Range )
4.2. Mercuric Sulphate
4.3. COD Standard ( Hach Cat. No.12186-49)
5. PROCEDURE
Turn ON the COD Reactor. Preheat to 150 °C .Mix the sample prior to homogenization.
Remove the cap of a COD Vial 0 to 150 ppm range. Hold the vial at a 45 –degree angle. Take
2.0 ml of sample into the vial using a pipette. Replace the vial cap tightly. Rinse the outside of
the COD vial with deionized water and wipe the vial. Invert gently several times to mix the
content. Place the vial in the preheated COD reactor. Prepare a blank by adding 2.0 ml of
deioned water to the COD Vial and place in the preheated COD vial. Heat the vials for two
hours. Turn the reactor off. Wait about 20 minutes for the vials to cool to 120 °C or less .Invert
each vial several times while still warm. Place the vials in to a rack. Wait until the vials have
cooled to room temperature.
GLAB-09 (Revised) - C.O.D. (HACH method) Page 2 of 3
Page 38
Power ON HACH spectrophotometer 2010. Call program # 430 and adjust the wave length to
420 nm. Place the COD vials adapter into the cell holder with the marker to the right. Clean
outside of the blank. Place the blank into the adapter .Place the cover on the adapter. Press
Zero.The display will show 0 mg/l COD LR. Place the sample vial into the adapter and cover
the adapter. Press READ. The display will show the result in mg/l COD.
Interferences
Chloride is the primary interference when determining COD .Each COD Vial (Low range 0 to
150 ppm) contains mercuric sulphate that will eliminate chloride interference upto the level of
2000 mg/l .By adding 0.50 g of mercuric sulphate to each vial the samples containing chloride
concentration up to 8000 mg/l can be tested.
Note: For Sea water sample, dilute the sample five times and do the analysis.
6. ACCURACY CHECK
Check the accuracy of the 0 to 150 mg/l range with Chemical oxygen demand standard solution
(Potassium acid phthalate ) Hach Cat. 12186-49
Prepare standard solution of 45 ppm standard solution from the stock solution of 300 ppm and
run the standard as sample . Apply correction factor for the sample if required.
7. SAFETY NOTE
7.1 Mercuric sulphate is very toxic, avoid contact with the chemical and its solutions.
7.2 Final sample will contain mercury, silver and chromium at concentration levels
regulated as hazardous waste. proper disposal of these materials.
Reference : ASTM D 1252

APHA 5220 (D)
GLAB-09 (Revised) - C.O.D. (HACH method) Page 3 of 3

Page 39
SECTION WATER
THE DETERMINATION OF CHEMICAL OXYGEN DEMAND

Page 40

CHEMICAL OXYGEN DEMAND
[COD Reflux METHOD]
G-LAB METHOD No.10
(K.P. Abraham)
1. SCOPE
This method covers the determination of the quantity of oxygen that certain impurities in water
will consume, based on the reduction of a dichromate solution under specified conditions. This
method can be applied to cooling waters, industrial effluents, domestic sewage etc. It can
measure COD values in the range of 50-800 mg/l. The measurement of COD may be used for
evaluating the organic contaminants and for evaluating the treatment and control of industrial
waste water.
2. PRINCIPLE
Most organic and oxidisable inorganic substances present in water are oxidized by a standard
potassium dichromate solution in 50 % sulphuric acid solution. The excess dichromate is
titrated with a standard ferrous ammonium sulphate solution using orthophenanthroline ferrous
complex as an internal indicator. The oxidation of many otherwise organics is facilitated by the
use of silver sulphate, which acts as a catalyst in the reaction. The oxidation of up to 1000 mg/l
of chloride ion is inhibited by the addition of mercuric sulphate to form a stable and soluble
mercuric chloride complex.
3. APPARATUS
3.1. Reflux apparatus
The apparatus consists of a 500 ml Erlenmeyer round bottom
flask made of heat resistant glass connected to a condenser by
means of a ground glass joint.
3.2. Heating apparatus
A heating mantle capable of delivering sufficient controlled
heat to maintain a steady reflux rate in the reflux apparatus.
3.4. Pipette, 10 ml.
3.5. Measuring cylinders, 10, 50 and 100 ml.
3.6. Beaker, 1 litre.
4. REAGENTS
4.1. Ferrous ammonium sulphate solution, standard (0.25 M).
Dissolve 98.0 g ferrous ammonium sulphate (FeSO4.(NH4)2SO4.6H2O) in water. Add
20 ml of concentrated sulphuric acid (H2SO4.Sp.gr 1.84), cool and dilute to 11itre.
Standardise this solution daily before use as per the method later.
4.2. Mercuric sulphate
Powdered mercuric sulphate (HgSO4).
4.3. Ferroin indicator
Dissolve 1.48 g of 1,10(ortho) phenanthroline monohydrate, together with 0.70 g of
GLAB-10 (Revised) - C.O.D. (Reflux method) Page 2 of 4

Page 41
FeSO4.7H2O, in 100 ml of distilled water.

4.4. Potassium acid phthalate solution, standard (lml =1 mg COD)
Dissolve 0.851 g of potassium acid phthalate (KC8H5O4) primary standard grade in
water and dilute to 1 litre.
4.5. Potassium dichromate solution, standard (0.0417M)
Dissolve 12.259 g of potassium dichromate (K2Cr2O7) primary standard grade,
previously dried at 103°C for 2 hours, in water and dilute to 1 litre in a volumetric
flask.
4.6. Silver sulphate
Powdered silver sulphate (Ag2SO4)
4.7. Sulphuric acid
Concentrated sulphuric acid (H2SO4 Sp.gr. 1.84)
4.8. Sulphuric acid Silver sulphate solution
Dissolve 10 g of powdered silver sulphate in 300 ml of concentrated sulphuric acid
and dilute to 1 litre with concentrated sulphuric acid,
5. STANDARDI ZATION
Standardisation of ferrous ammonium sulphate solution
Pipette 10.0 ml of the standard potassium dichromate solution (0.04l7M) to 250 ml Erlenmeyer
flask and dilute to about 100 ml with distilled water. Add 30 ml concentrated sulphuric acid
and cool. Titrate with the ferrous ammonium sulphate solution using ferroin indicator
(3 drops). The end point is noticed by the colour change from blue green to reddish brown.
Note the titre reading.
0.04l7M K2Cr2O7 = 0.25N K2Cr2O7
Normality, N = 10 x 0.25
Titre reading
6. PROCEDURE
Preserve samples by cooling to 4° C if analysed within 24 hours after sampling, or preserve for
up to 7 days at pH <2 at 4°C. The addition of 2 ml of concentrated H2SO4 per litre at the time
of collection will generally achieve this requirement.
Shake the sample well and take 50 ml sample in a reflux flask. Place the reflux flask in an ice
bath and add 1 gm of powdered mercuric sulphate HgSO4, 5.0 ml of concentrated Sulphuric
acid (H2SO4) and several glass beads. Mix well to complete dissolution. With the flask still in
the ice bath add slowly and with stirring 10.0 ml of 0.25 N standard K2Cr2O7 solution. Attach
the flask to the condenser and start the flow of cold water.
Add 70 ml of sulphuric acid-silver sulphate solution slowly through the open end of the
condenser, continue swirling the flask while the acid mixture is being added. Apply heat to the
flask and reflux for 2 hours. Allow the flask to cool and wash down the condenser with distilled
water. Add 3 drops of the ferroin indicator and titrate with the standardised ammonium ferrous
sulphate solution. The end point is noted by the colour changing from blue green to reddish
brown. (If the solution immediately turns a reddish brown upon the addition of the indicator
repeat the analysis on a smaller sample aliquot). Note the titre reading as ‘B’.
Place 50 ml distilled water in a reflux flask and follow the same procedure used for the sample,
for the blank determination. Let the titre reading be A’.

Page 42
To check the validity of the test result, make a standard determination using potassium acid
phthalate solution. A COD of 100 mg/litre should be obtained on a 5 ml aliquot of the standard
solution (l ml = 1mg COD) diluted to 50 ml.
7. CALCULATION
Chemical oxygen demand mg/litre = (A - B)N x 8000
V
Where,
A = millilitres of ferrous ammonium sulphate solution required for the titration of the blank.
B = millilitres of ferrous ammonium sulphate solution required for the titration of the sample.
N = normality of the ferrous ammonium sulphate solution.
V = millilitres of sample used for the test.
8. SAFETY NOTE
8.1 Excercise extreme care when handling concentrated sulphuric acid.

8.2 Silver sulphate is poisonous, avoid contact with the chemical and its solution.
8.3 Mercuric sulphate is very toxic, avoid contact with the chemical and its solutions.

APHA 5220 (B)

Page 43
SECTION: WATER
THE DETERMINATION OF CHEMICAL TANK CONCENTRATION

(REFRACTOMETER METHOD)

Prepared by : K. P. Abraham
Page 44

CHEMICAL TANK CONCENTRATION
(REFRACTOMETER METHOD)
G-LAB METHOD No.11
(By K.P. Abraham)
1. SCOPE
This method covers the determination of the concentration of chemicals in percentage.
Different chemicals, which are prepared on site for dosing purposes, can be checked by this
method, eg. Inhibitors for the desalination plants, trisodium phosphate for the boilers. This is a
quick field method modified to suit our plant operations.
2. PRINCIPLE
The refractive index of the liquid under test is determined using the principle of the critical
angle of refraction, which is proportional to the concentration of the solution. The scale of the
refractometer is made so that it can indicate the correct value when the refractometer is used for
measurement at a specified temperature.
3. APPARATUS
3.1. Hand held refractometer
ATAGO Model N 10.
3.2. Volumetric flask, 100 ml (5 Nos.)
3.3. Analytical balance.
3.4. Beakers, 500 ml.
3.5. Thermometer Range 0-50° C.
4. REAGENTS
4.1. Distilled water
Distilled water at 20’C.
4.2. Standard solutions

Weigh accurately 1.000, 2.000, 3.000, 4.000 and 5,000 g of the chemical which is to
be checked in clean 50 ml beaker and add 25 ml distilled water. Transfer
quantitatively to 100 ml volumetric flask. Make up to the mark with distilled water at
20°C. This will give standard solutions of the chemicals with 1%, 2%, 3%, 4% and
5% w/v concentrations.
GLAB-11 (Revised) - Concentration by Refractometer Page 2 of 3

Page 45
5. CALIBRATION
Put a few drops of water on the prism close the daylight plate gently. Check whether the water
has spread all over the prism surface. Look at the scale through the eyepiece. To have clear
view, turn focusing ring. Read the scale where the boundary line intercepts it. Adjust reading to
zero with the help of zero adjusting screw. (Turning anticlockwise makes the level move up).
After adjusting zero, wipe and clean the prism and daylight plate surface with tissue paper. Put
few drops of the first standard (1%) on the prism. Close daylight plate gently. Check whether
the solution has spread all over the prism surface. Look at the scale through the eye piece. Read
the scale where the boundary line intercepts and note the reading. Repeat this with all the
standards and note the readings. Plot a graph of concentration in percentage versus the scale
readings. Calculate a factor, which is equal to concentration in percentage for unit scale
reading.
6. PROCEDURE
Collect sample from the chemical tank and the water, which is used for making the chemical
solution. Keep at room temperature for a period. Check the temperature of the sample and the
make up water to make sure both are the same. Put few drops of the make up water on the
prism and adjust the reading to zero as described in the calibration procedure. Then wipe and
clean the prism and daylight plate surface with tissue paper. Put few drops of the sample on the
prism. Check the scale reading as described earlier. Note the refractometer scale reading for the
sample.
7. CALCULATION
Chemical concentration, % = Refractometer reading x F
Where,
F = Factor from the calibration graph.
(Note: The factor ‘F’ will be different for different chemicals).
Reference: Instruction manual for Hand held Refractometer.
GLAB-11 (Revised) - Concentration by Refractometer Page 3 of 3

Page 46
SECTION: WATER
(SPECIFIC GRAVITY BOTTLE METHOD)

Page 47

(SPECIFIC GRAVITY METHOD)
G-LAB METHOD No.12
(By K.P. Abraham)
1. SCOPE
This method covers the determination of the concentration of chemicals in percentages.
Different chemicals, which are prepared on site for dosing purposes, can be checked by this
method. (e.g., different inhibitors used in the desalination plant). This is a modified method to
suit our plant operations and it is more accurate than the refractometer method.
2. PRINCIPLE
This method is based on the relation between the specific gravity of a solution and its
concentration. The specific gravity at a specified temperature increase with the concentration,
there by the weight of a certain volume of the sample at the same temperature will be
proportional to its concentration. Here a modified technique of this is used i.e. the relation
between the weight difference of a certain volume of the sample at the same temperature and
its concentration.
3. APPARATUS
3.1. Specific gravity bottle, 50 ml
3.2. Analytical balance
3.4. Thermometer, 0 to 50°C
4. REAGENTS
4.1. Distilled water Distilled water at 20°C.

Weigh accurately 1.000, 2.000, 3.000, 4.000 and 5.000 g of the inhibitor in 50 ml
clean beakers and add 25 ml distilled water. Transfer quantitatively to 100 ml
volumetric flasks and make up to the mark with distilled water at 20°C. This will give
standard solutions of the inhibitor with 1%, 2%, 3%, 4% and 5% w/v concentrations.
GLAB-12 (Revised) - Concentration by Sp.Gravity Page 2 of 3

Page 48
5. CALIBRATION
5.1. Calibration and Standardization
Check the temperature of the blank distilled water and the standard solutions. Allow
them to attain room temperature. Fill the specific gravity bottle with distilled water
and fit the stopper. Allow the excess water to flow out through the hole in the stopper.
Wipe the outside of the bottle with tissue paper and remove all excess water. Weigh
the sp. gravity bottle with distilled water in an analytical balance. Note the weight.
Empty the bottle and rinse it with the first standard (1%) and fill. Fill stopper, wipe
the outside with tissue paper and weigh. Repeat the same procedure with all the
standards. Note their respective weights. Calculate the weight difference between the
specific gravity bottle with distilled water blank and the specific gravity bottle with
the standard solutions. Plot a graph of concentration in percentage versus the weight
difference in grams. Calculate a factor from the graph, which is equal to concentration
in percentage for a unit weight difference in grams.
6. PROCEDURE
Collect sample from the chemical tank and the water which is used for making the
chemical solution. Keep at room temperature for a period. Check the temperature of
the sample and the make up water to ensure both are the same. Fill the specific
gravity bottle with the make up water. Follow the same procedure used for the
standards, for calibration, and weigh the specific gravity bottle with the blank water.
Let it be W1 g. Empty the bottle and fill with the sample after rinsing. Weigh as
before. Let the weight be W2 g.
7. CALCULATIONS
Chemical concentration, % w/v = (W2-W1) x Factor from calibration graph
Where,
W1 = weight of sp.gr. bottle + blank, g
W2 = weight of sp.gr. bottle + sample, g
GLAB-12 (Revised) - Concentration by Sp.Gravity Page 3 of 3

Page 49
SECTION: WATER

(DENSITY BALANCE METHOD)
Method No : GLAB -13

Date issued : 5th November 1995
Prepared by : K.P.Abraham
Page 50

G-LAB METHOD No.13
(By K.P. Abraham)
1. SCOPE
This method covers the determination of the concentration of chemicals in percentages.

Different chemicals which are prepared on site for dosing purposes can be checked by this
method. (e.g., different inhibitors used in the desalination plant). This is a modified method to
suit our plant operations.
2. PRINCIPLE
This method is based on the relation between the density of a solution and its concentration.
The density at a specified temperature increases with the concentration, there by the weight of a
certain volume of the sample at the same temperature will be proportional to its concentration.
3. APPARATUS
3.1. Density balance
3.2. Measuring jar
3.4 Thermometer, 0 to 50°C
4. REAGENTS

Distilled water at 20°C.

Weigh accurately 2.000, 4.000, 6.000, 8.000 and 10.000 g of the inhibitor in .50 ml
clean beakers and add 25 ml distilled water. Transfer quantitatively to 100 ml
volumetric flasks and make up to the mark with distilled water at 20°C. This will give
standard solutions of the inhibitor with 2%, 4%, 6%, 8% and 10% w/v concentrations.
5. CALIBRATION
Zero point calibration

Carefully clean and dry the plummet and hang it on the hook. Keep both the sliding
weights on the 0.000 positions. Keep the beam weight in position. Turn the screws on
the foot and at the left of the beam to zero the balance.
Standardisation
GLAB-13 (Revised) - Concentration by Density Balance Page 2 of 3
Page 51
Fill up the measuring jar 20 mm below the rim with the first standard and
immerse the plummet into it.
Adjust the sliding weight and make the balance steady if the density of the
standard is more than 1 then remove beam weight to steady the balance.
Read the weight by adding the slider weights. Add 1 if the hanging weight is
off the beam.
The reading of the balance is corrected according to the correction schedule,

because air buoyancy and liquid bulge at the wire of the plummet, are going
into the measurement.
CORRECTION SCHEDULE
Density Correction Density Correction

0.6000 +0.0003 1.4000 -0.0006
0.7000 +0.0002 1.5000 -0.0008
0.8000 +0.0001 1.6000 -0.0010
0.9000 0.0000 1.7000 -0.0011
1.0000 0.0000 1.8000 -0.0013
1.1000 -0.0002 1.9000 -0.0015
1.2000 -0.0003 2.0000 -0.0017
1.3000 -0.0005
Repeat the same procedure with all the standards. Note their respective densities
Plot a graph of concentration in percentage versus the corresponding density

of the standard.
6. PROCEDURE
6.1. Carefully clean and dry the plummet and hang it on the hook. Keep both the
sliding weights on the 0.000 positions. Keep the beam weight in position. Turn
the screws on the foot and at the left of the beam to zero the balance.
6.2. Pill up the measuring jar 20 mm- below the rim with sample and immerse the
plummet into it.
6.3. Adjust the sliding weight and make the balance steady. If the density of the
standard is more than 1 then remove beam weight to steady the balance.
6.4. Read the weight by adding the slider weights. Add 1 if the hanging weight is
off the beam.
6.5. The reading of the balance is corrected according to the correction schedule
because, air buoyancy and liquid bulge at the wire of the plummet, are going
into the measurement.
7. CALCULATIONS
Chemical concentration, % w/v to be read from the graph.
GLAB-13 (Revised) - Concentration by Density Balance Page 3 of 3

Page 52
G – STATION LABORATORY METHOD
SECTION: WATER
THE DETERMINATION OF CHLORIDE (LOW RANGE)

Page 53

CHLORIDE (LOW RANGE)
G - LAB METHOD No.14
(By K.P.Abraham)
1. SCOPE
This method covers thee determination of chloride in boiler feed water, boiler water, distilled
water; distillate etc., where it will be in the range of 1 to 20 mg/l. Iron in excess of 10 mg/l
interferes by masking the end point.
2. PRINCIPLE
Water sample adjusted to pH 8.3 is titrated with silver nitrate solution in the presence of
potassium chromate indicator. The end point is indicated by persistence of the pinkish yellow
colour.
3. APPARATUS
3.1. Automatic micro burette, 10 ml

3.3. Conical flask, 250 ml
3.4. Indicator bottles
3.5. Volumetric flask, 1 lit
3.6. Droppers
3.7. Weighing balance.
4. REAGENTS
4.1. Phenolphthalein indicator solution

Dissolve 1 g of phenolphthalein in 60 ml of alcohol and then add 40 nil of distilled
water and mix.

Distilled water with conductivity less than 1 µS/cm.
4.3. Potassium chromate indicator — solution

Dissolve 50 g of potassium chromate (K2CrO4) in 100 ml of distilled water, and add
silver nitrate (AgNO3) until a slight red precipitate is produced. Allow the solution to
stand, protect from sunlight, for atleast 24 hours after the addition of AgNO3. Then
filter the solution to remove the precipitate and dilute to 1 litre with distilled water.
4.4. Silver nitrate solution, standard (0.0141M)

Crush approximately 5 g of silver nitrate (AgNO3) crystals and dry to constant weight
at 40C. Dissolve 2.3950 + 0.0002 g of the crushed, dried crystals in distilled water
and dilute to 1 litre. Standardize against the standard NaCl solution.
GLAB-14 (Revised) - Chloride (Low range) Page 2 of 4

Page 54
4.5. Sodium chloride solution, standard (0.0141M)

Dry several grams of sodium chloride (NaCl) for 1 hour at 600°C. Dissolve 0.8240 +
0.0002 g of the dry salt in distilled water and dilute to 1 litre at 25°C in a volumetirc
flask.
4.6. Sodium hydroxide solution (10 g/l)

Dissolve 10 g of sodium hydroxide (NaOH) in distilled water and dilute to 1 litre.
4.7. Sulphuric acid (1+19)

Carefully add 1 volume of concentrated sulphuric acid (H2S04 sp. gravity 1.84) to 19
volumes of water, while mixing.
5. CALIBRATION
Standardization of Silver nitrate solution
Pipette 10.0 ml of the standard sodium chloride solution (0.0141M NaCl) to a 250 ml conical
flask and add 90 ml blank distilled water. Add 1.0 ml potassium chromate indicator solution.
Titrate with the silver nitrate solution from the burette to a pinkish yellow end point. Note the
titre reading as V1 ml.
Take 100 ml blank distilled water in a 250 ml conical flask. Add 1.0 ml potassium chromate
indicator and titrate against the silver nitrate. Note the titre reading V2, ml at the end point. A
blank reading of 0.2 to 0.3 ml is usual.
Normality of silver nitrate = 0.0141x10

(V1 — V2)
Where,
V1 = Titre reading for sodium chloride solution, ml.

V2 = Titre reading for the blank distilled water, ml.
0.0141 = Normality of sodium chloride
(0.0141M NaCl = 0.0141N NaC1)
10 = Standard NaC1 volume, ml
6. PROCEDURE
Pour 100 ml of the sample into a 250 ml conical flask. Adjust the pH to phenolphthalein end
point (pH 8.3), using H2SO4 (1 + 19) or NaOH solution 10 g/l) with 1 drop of phenolphthalein
indicator.
Add approximately 1.0 ml of K2CrO4 indicator solution and mix. Add standard AgNO3 solution
dropwise from a 10 ml automatic micro burette until the pinkish yellow end point. Note the
burette reading as ‘V 3,’ ml.
Repeat the above procedure with 100 ml distilled water and note the burette reading as ‘V4’,
ml.

Page 55
7. CALCULATION
Chloride, mg/I = (V4 —V3) x 35.46 x 0.0141 x 1000

Sample volume, ml
Where,
V3 = Sample titre reading, ml.
V4 = Blank titre reading, ml.
35.46 = Equivalent weight of chlorine.
0.0141 = Normality of silver nitrate.
Reference : APHA 4500 Cl- (B)

ASTM D 512 – 89 (Reapproved 1994)

Page 56
G - STATION LABORATORY METHOD
SECTION: WATER
THE DETERMINATION OF CHLORIDE (VERY LOW RANGE)

Page 57

CHLORIDE (VERY LOW RANGE)
G-LAB METHOD No.15
(By K.P.Abraham)
1. SCOPE
This method is applicable to the determination of very low levels of chlorides in boiler feed
water, makeup water, demineralised water, steam condensate, boiler water, etc. This method is
specifically applicable to chloride levels of 0.05 to 5.0 ppm.
2. PRINCIPLE
The method is based on the measurement of the turbidity formed by the formation of silver
chloride in the solution. The intensity is measured photometrically at 420 nm wave length.
3. APPARATUS
3.1 Spectrophotometer
JASCO Model V 530
3.2 Sample cell, 50 mm.
3.3 Beakers, 100 ml (10 nos.)
3.4 Glass rod.
3.5 Measuring cylinder, 100 ml.
3.6 Volumetric flask, 100 ml, 1 lit.
3.7 Weighing balance.
3.8 Water polishing unit, MilliQ system
3.9 Pipettes, 1, 2, 3, 5, 10 ml
3.10 Beaker, 500 ml.
4. REAGENTS
4.1 Demineralized Water

Use water from the water polishing unit.
4.2 Nitric Acid (1 + 2)
Take 200 ml demineralized water in a 500 ml beaker. Measure 100 ml con. Nitric
acid (sp. gravity) and mix with the water.
4.3 Silver Nitrate (0.5% solution)
Dissolve 0.5 g of Silver Nitrate (AgNO3) in demineralized water and make up to
100 ml in a volumetric flask.
4.4 Chloride Solution, standard (1 ml = 1 mg Cl)
Dry about 5 g of Sodium Chloride (NaCl) at 180°C for 2 hours. Dissolve 1.6481 g
of the dried Sodium Chloride in demineralized water and make up to 1 litre in
volumetric flask.
GLAB-15 (Revised) - Chloride (Turbidity method) Page 2 of 3
Page 58
4.5 Chloride Solution, Standard (1 ml = .01 mg Cl)

Pipette 10.0 ml of the standard Chloride solution (1 ml 1 mg Cl) to a 1 litre
volumetric flask and make up to the mark with demineralized water.
5. CALIBRATION
Pipette 1, 2, 3, 4, 5, 10, 15, 20, 25, & 30 ml of the standard Chloride solution (1 ml = 0.01 mg
Cl) to 100 ml beakers and add the remaining volume of de-ionized water to make the volume
as 50 ml. This will provide standards of 0.2, 0.4, 0.6, 0.8, 1.0, 2.0, 3.0, 4.0& 5.0 mg/l. Add 5
ml of Nitric acid (1 + 2) to each and mix well. Add 1 ml of silver nitrate solution to each and
mix well. Take 50 ml de-ionized water and add 5 ml Nitric acid (1 + 2) and mix thoroughly and
use this as the blank. Wait for 10 minutes. Measure the absorbance at 420 nm wave length
using 50 mm cells in Spectrophotometer JASCO V 530 and save the calibration graph.
6. PROCEDURE
Take 50 ml sample in 100 ml beaker. Add 5 ml Nitric Acid solution and mix well. Add 1 ml
Silver Nitrate solution and mix well. Take 50 ml de-ionized water in another beaker and add 5
ml Nitric Acid solution and mix well. Wait for 10 minutes. Measure the absorbance at 420 nm
wavelength using 50 mm cell.
In G—Station laboratory, use method No.4 in spectrophotometer for chloride in the range of
0.05 to 5 ppm. By calling the specified method number all the parameters required for the
analysis will be set in the instrument and the concentration can be directly measured.
7. CALCULATION
Use the corresponding calibration graphs and read the concentration of chloride against the
absorbance. Report the concentration of Chloride as mg/l (ppm).
GLAB-15 (Revised) - Chloride (Turbidity method) Page 3 of 3

Page 59
SECTION: WATER
THE DETERMINATION OF CHLORIDE IN WATER (HIGH RANGE)

Page 60

CHLORIDE (HIGH RANGE)
G-LAB METHOD No.16
(By K,P.Abraham)
1. SCOPE
This method covers the determination of chloride in drinking water, cooling water,
waste water and saline water etc., where the chloride levels will be >50 mg/l.
2. PRINCIPLE
The estimation of chloride is based on the titration of the sample with standard silver nitrate
solution using potassium chromate as indicator, The end point is indicated by persistence of the
brick-red silver chromate colour.
3. APPARATUS
3.1. Automatic burette, 25 ml.
3.2. Measuring cylinder, 100 ml.
3.3. Pipettes, 1, 5, 10 and 25 ml.
3.4. Conical flask, 250 ml.
3.5. Volumetric flask, 1 lit.
3.6. Indicator bottle.
4. REAGENTS
4.1. Potassium chromate indicator solution

Dissolve 5 g of potassium chromate (K2CrO4) in 100 ml distilled water.
4.2. Silver nitrate solution, standard (0.141 M)

Crush approximately 25 g of silver nitrate (AgNO3) crystals and dry to constant
weight at 40°C. Dissolve 23.954 + 0.001 g of the crushed, dried, crystals in
distilled water and dilute to 1litre distilled water. Standardise with standard NaC1
solution (0.141 N).
4.3. Silver nitrate solution, standard (0.282 M)

weight at 40°C. Dissolve 47.908 + 0.001 g of the crushed, dried, crystals in
distilled water and dilute to l litre with distilled water. Standardise with standard
NaCl solution (0.141 N).
4.4. Sodium chloride solution, standard (0.141 M)

Dry about 10 g of sodium chloride (NaCl) for 1 hour at 140°C. Dissolve 8.2417 +
0.0002 g of the dry salt in distilled water and dilute to l litre 25 °C in a volumetric
flask.
GLAB-16 (Revised) - Chloride (High range) Page 2 of 3
Page 61
5. STANDARDIZATION
Pipette 10.0 ml of the standard sodium chloride solution (0.141 M) in a 250 ml conical flask.
Add 5 drops of potassium chromate indicator and mix titrate against the silver nitrate solution
from a burette, Note the end point when the brick-red colour persists.
0.141 M NaC1 = 0.141 N NaCl

Normality of AgNO3 = 10 x 0.141
Titre reading, ml
If the normality of the silver nitrate difference from 0.141/0.282, then adjust the normality to
0.141/0.282 by adding calculated volume of distilled water or calculated quantity of silver
nitrate. Standaridize again, after the adjustments, with same procedure given earlier.
0.141 N AgNO3 = 0.141 M AgNO3
0.282 N AgNO3 = 0.282 M AgNO3
6. PROCEDURE
Take samples as follows

100 ml if conductivity < 700 µS/cm
50 ml if conductivity >700 but <1500 µS/cm
5 ml if the sample in a saline water (eg. sea water, brine recycle etc).
The remaining portion, distilled water to be added to make the volume about 100 ml. Add 5
drops of K2CrO4 indicator and mix. Titrate against the standard AgNO3 solution, 0.141 M
incase of normal water but 0.282 M incase of saline water, until the brick-red colour persists.
Note the titre reading, ml.
7. CALCULATION
If 0.141 M AgNO3 is used,
Chloride mg/l = Titre reading x 35.46 x 0.141 x 1000

Sample volume

Sample volume
If 0.282 M AgNO3 is used,
Chloride mg/l = Titre reading x 35.46 x 0.282 x 1000

Sample volume
Sample volume

: APHA 4500-Cl- (B)
GLAB-16 (Revised) - Chloride (High range) Page 3 of 3

Page 62
SECTION: WATER
THE DETERMINATION OF CHLORINE (FREE AND TOTAL)

Page 63

FREE CHLORINE AND TOTAL CHLORINE
G-LAB METHOD No.17
(By K.P.Abraham)
1. SCOPE
This test method covers the determination of free chlorine and total chlorine present in
chlorinated waters such as drinking water, cooling water, waste water, sea water etc. This
covers a range of free Cl2 0 - 2 mg/l. For the determination two methods are used.
METHOD - A
2. PRINCIPLE
Chlorine applied to water in its elemental or hypochlorite form initially undergoes hydrolysis to
form free chlorine consisting of aqueous molecular chlorine, hypochlorous acid and
hypochlorite ion. The relative proportion of these free chlorine forms is pH and temperature
dependent. In the absence of iodide ion, free chlorine reacts instantly with N,N-diethyl-P-
phenylenediamine (DPD) indicator to produce a red colour. The intensity of the colour is
proportional to the chlorine concentration.
3. APPARATUS
3.1. Lovibond comparator
Lovibond 2000 Comparator with integrated prism.
3.2. Cells — 13.5 mm moulded cells DB 424
3.3. Colour discs Colour disc code 3/40A
3.4. Tablet crusher
4. REAGENTS
4.1 DPD tablets No.1 and No.4
4.2 Lovibond Comparator DPD No.1 and DPD No.4.
5. PROCEDURE
Open the lovibond comparator and insert the colour disc (0-1.0 mg/l 012). Take two cells, fill
one cell upto the mark with the sample. Wipe the cells to remove any traces of water. Insert this
to the left side compartment. Fill the other cell with the sample upto the mark. Add the DPD
No.1 tablet. Crush the tablet using the tablet crusher. Mix well and insert this in the right side
compartment. Then see through the integrated prism and compare the colour on both sides.
Bring the left side colour near to the right side by rotating the colour disc. Take the reading
shown at the side of the comparator, when the colour matches. For measuring total chlorine use
DPD No.4 tablet instead of DPD No.1, wait for one minute.
6. CALCULATION
Free chlorine, mg/l = Reading obtained from the compactor disc (when DPD No.1 tablet is
used).
GLAB-17 (Revised) - Chlorine (Free & Total) Page 2 of 4
Page 64
Total chlorine, mg/l = Reading obtained from the compactor disc (when DPD No.4
tablet is used).
Reference : APHA 4500 Cl (G), Manual of Lovibond Comparator.
METHOD - B
2. PRINCIPLE
Chlorine reacts instantly with N,N-diethyl-P-phenylenediamine (DPD) indicator to produce a
red colour. The intensity of the colour is proportional to the chlorine concentration. Since
chlorine also can produce similar colour with DPD, chlorine to be eliminated before proceeding
with ClO2 determination. Chlorine is eliminated by the addition of Glycine.
3. APPARATUS
3.1 HACH Pocket colorimeter
3.2 Cylinder, graduated mixing, 50 ml
3.3 Pipet, volumetric, 1.00 ml
3.4 Pipet filler, safety bulb
3.5 Samples cells 10.0 ml, HACH Cat.No.24276-06
4. REAGENTS
4.1 DPD Free Chlorine reagent powder pillows, HACH Cat. No. 21055-69
4.2 DPD Total Chlorine reagent powder pillows, HACH Cat. No. 21056-69
5. PROCEDURE
5.1 Fill a 10 ml cell to the 10 ml line with sample (the blank). Close with the cap and
wipe off sample cells.
5.2 Remove the instrument cap and place the blank in the cell holder, with the diamond
mark facing you. Tightly cover the cell with the instrument cap (flat side facing the
back of the instrument).
5.3 Press ZERO. The instrument will turn on and the display will show --- the 0.00
5.4 Remove the cell from the cell holder.
5.5 Fill a 10 ml cell to the 10 ml line with sample.
5.6 Add the contents of one DPD chlorine powder pillow to the sample cell (the
prepared sample). Cap and shake gently for 20 seconds. Wipe liquid off sample
cell. A pink colour will develop if chlorine is present.
5.7 Within 1 minute after adding DPD (free chlorine) to the sample or after 3 minutes
after adding DPD (total chlorine) to the sample, place the prepared sample in the
cell holder.
5.8 Tightly cove the cell with the instrument cap (flat side facing the back of the
instrument).
5.9 Press READ. The instrument will show --- followed by the results in mg/L free
chlorine.
5.10 Note the displayed reading.

Page 65
6. CALCULATION
Free Chlorine, mg/l = Reading obtained from the colorimeter when DPD free chlorine reagent
(DPD No.1) is used
Total Chlorine, mg/l = Reading obtained from the colorimeter when DPD total chlorine
reagent (DPD No.4) is used
Reference: APHA 4500 Cl (G), Manual of HACH pocket colorimeter.

Page 66
SECTION: WATER
THE DETERMINATION OF TOTAL AVAILABLE CHLORINE

Page 67
TEST METHOD FOR HYPOCHLORATES

TOTAL AVAILABLE CHLORINE
G-LAB METHOD No.18
(By K.P.Abraham)
1. SCOPE
This test method is used for the determination of the total available chlorine in bleaching
powder and hypochlorite solutions. The colourimetric end point may be obscured by
excessive turbidity and colour, which will limit the required precision to visually observe
the end point.
2. PRINCIPLE
A definite weight of the sample, suspended in water, or a definite volume of the sample, if
liquid, is treated with potassium iodide in acid medium. An equivalent amount of iodine is
liberated, which is titrated against the standard sodium thiosulphate solution, using starch as
indicator.
3. APPARATUS
3.3. Pipette, 10 ml
3.6. Volumetric flasks, 100 ml, 1 litre
4. REAGENTS
4.1. Sodium thiosulphate, Standard (0.1M)

Dissolve 24.82 g of sodium thiosulphate (Na2S2O3.5H2O) (15.82 gms of
anhydrous Na2S2O3) in boiled and cooled distilled water. Dilute to 1L and add 0.1
g of sodium carbonate as a preservative. This solution deteriorates on standing and
should be standardised every two weeks, using standard iodine solution.
4.2. Iodine solution, Standard (0.05M)
To be prepared from a BDH CVS (Prod. No. 18038) or equivalent. Transfer
quantitatively to a 1 litre volumetric flask and make up to the mark with distilled
water.
4.3. Potassium iodide - 10%
Dissolve 10 g of KI in distilled water and dilute to 100 ml.
4.4. Acetic acid - 30%
Add 30 ml glacial acetic acid to 50 ml distilled water and make up to 100 ml with
distilled water.
0.5 g of soluble starch is made to a thin paste with a little cold water and then add
25 ml. of boiling water. Boil until a clean solution is obtained. This should be
freshly prepared as required.
GLAB-18 (Revised) - Chlorine (Total available) Page 2 of 3

Page 68
5. CALIBRATION
Standardization of sodium thiosulphate solution
Pipette 10.0 ml of the standard iodine solution (0.05M) to a 250 ml conical flask and make
up to about 100 ml with distilled water. Take the sodium thiosulphate solution in the
burette. Titrate the iodine solution with the thiosulphate solution. When the colour of the
solution changes to pale yellow from the initial colour, add 1 ml of starch indicator.
Continue the titration. The end point is noted when the colour changes from blue to
colourless. Note the titre reading as V1, ml.
Normality of sodium thiosulphate = 0.1 x 10

V1
Where,
V1 = Titre reading, ml
0.1 = Normality of iodine solution (0.05M iodine = 0.1N iodine)
6. PROCEDURE
Pipette 10 ml of the sample into a 250 ml conical flask and make up to about 100 ml with
distilled water. Add 5 ml. of acetic acid (30%) solution and mix. Then add 5 ml. of
potassium iodide (10%) solution and mix. Titrate with the standard sodium thiosulphate
solution. Add 1 ml. of starch indicator towards the end of the titration. At the end point,
colour changes from blue to colourless. Note the titre reading V2, ml.
Note: For concentrated hypochlorite samples take 1ml of the sample only.
7. CALCULATION
Total available chlorine, mg/l as Cl2 = V2 x 35.46 x N x 1000

Sample volume
Where,
V2 = Titre reading for sample, ml

N = Normality of sodium thiosulphate
35.46 = Equivalent weight of Chlorine.
Note: For concentrated hypochlorite samples, report in % w/v
Total available chlorine, %w/v as Cl2 = __V2 x 35.46 x N__

Sample volume x 10
Reference APHA 4500 Cl (B)
GLAB-18 (Revised) - Chlorine (Total available) Page 3 of 3

Page 69
SECTION: WATER
THE DETERMINATION OF CHLORINE DIOXIDE

Page 70

CHLORINE DIOXIDE (DPD METHOD)
G-LAB METHOD No.19
(K.P. Abraham)
1. SCOPE
This method covers the determination of chlorine dioxide in drinking water.Chlorine dioxide is
a strong oxidizing agent and is unstable in natural waters. It reacts rapidly with various
inorganic compounds, but oxidizes organic compounds more slowly. Many factors, including
reactant concentrations, sunlight, pH, temperature, and salinity influence decomposition of
chlorine dioxide in water. This test method covers the determination chlorine dioxide present in
drinking water. This covers a range of free ClO2 0 - 1 mg/l
2. PRINCIPLE
Chlorine dioxide reacts instantly with N,N-diethyl-P-phenylenediamine (DPD) indicator to
produce a red colour. The intensity of the colour is proportional to the chlorine concentration.
Since chlorine also can produce similar colour with DPD, chlorine to be eliminated before
proceeding with ClO2 determination. Chlorine is eliminated by the addition of Glycine.
3. APPARATUS
3.1 HACH Pocket colorimeter

3.5 Samples cells 10.0 ml, HACH Cat.No.24276-06
4. REAGENTS
4.1 DPD Free Chlorine reagent powder pillows, HACH Cat. No. 21055-69
4.2 Glycine tablets
5. PROCEDURE
5.1 Fill a 10 ml cell to the 10 ml line with sample (the blank). Close with the cap and
wipe off sample cells.
5.2 Remove the instrument cap and place the blank in the cell holder, with the diamond
mark facing you. Tightly cover the cell with the instrument cap (flat side facing the
back of the instrument).
5.3 Press ZERO. The instrument will turn on and the display will show --- the 0.00
5.4 Remove the cell from the cell holder.
5.5 Fill a 10 ml cell to the 10 ml line with sample. (Note: If the sample contains
chlorine, add one Glycine tablet to each sample and crush it to dissolve in it).
5.6 Add the contents of one DPD free chlorine powder pillow to the sample cell (the
prepared sample). Cap and shake gently for 20 seconds. Wipe liquid off sample
GLAB-19 (Revised) - Chlorine Dioxide (DPD method) Page 2 of 3

Page 71
cell. A pink colour will develop if ClO2 is present.

5.7 Within 1 minute after adding DPD to the sample, place the prepared sample in the
cell holder.
5.8 Tightly cove the cell with the instrument cap (flat side facing the back of the
instrument).
5.9 Press READ. The instrument will show --- followed by the results in mg/L free
chlorine.
5.10 Note the displayed reading.
6. CALCULATION
Chlorine dioxide, mg/l = Reading obtained from the colorimeter X 1.5
Reference: Manual of HACH pocket colorimeter.
GLAB-19 (Revised) - Chlorine Dioxide (DPD method) Page 3 of 3

Page 72
SECTION WATER
THE DETERMINATION OF CHLORINE DIOXIDE

Date issued : 01st June 2008
Page 73

CHLORINE DIOXIDE (HACH METHOD)
G-LAB METHOD No.20
(By K.P. Abraham & Philip Varky)
1. SCOPE
This method covers the determination of chlorine dioxide in drinking water.Chlorine dioxide is
a strong oxidizing agent and is unstable in natural waters. It reacts rapidly with various
inorganic compounds, but oxidizes organic compounds more slowly. Many factors, including
reactant concentrations, sunlight, pH, temperature, and salinity influence decomposition of
chlorine dioxide in water.
2. PRINCIPLE
Chlorine dioxide (ClO2) is determined by its combination with chlorophenol red at pH 5.2 to
from a colorless complex. The net effect of bleaching of the colour in an amount proportional
to the chlorine dioxide concentration. The method is specific for Chlorine dioxide and is
uncreative to other active chlorine or moderate oxidizing compound. Test tests are measured at
575 nm.
3. APPARATUS
3.1 HACH Spectrophotometer 2010

3.5 Samples cells 25.0 ml
4. REAGENTS
4.1 Chlorine Dioxide Reagent # 1 Hach Cat # 20700-42

4.4 Dechlorinating Reagent Power Pillows Hach Cat # 14363-69
5. PROCEDURE
5.1 Fill two 50 ml mixing cylinders to the 50 ml mark with sample. One is Blank and other
sample.
5.2 Use a volumetric pipette and pipette filler to add 1.0 ml of chlorine dioxide reagent 1 to
each cylinder. Stopper. Invert several times to mix.
5.3 To one cylinder, add one Dechlorinating reagent power pillow. (This will be Blank)
5.4 Use a volumetric pipette to add exactly 1.0ml of chlorine dioxide reagent # 2 to each
cylinder. Stopper and invert several times to mix.
GLAB-20 (New) - Chlorine Dioxide (HACH method) Page 2 of 3

Page 74
5.5 Use a volumetric ppipet and pipet filler to add 1.0 ml of chlorine dioxide reagent # 3 to
each cylinder stopper and invert several times to mix.
5.6 Power ON the spectrophotometer and press program # 72. Dial the wavelenth to 575
nm.
5.7 Pour 25 ml from each cylinder into square sample cell.
5.8 Pace the blank into the cell holder. Close the light shield. Press Zero The display will
show 0.00 m/l CLO2 LR
5.9 Place the prepared sample into the cell holder. Press READ. The display will show the
result in mg/l ClO2
6. CALCULATION
Chlorine dioxide, mg/l = Reading displayed on the spectrophotometer.
Reference: Manual of HACH DR 2010 spectrophotometer.
GLAB-20 (New) - Chlorine Dioxide (HACH method) Page 3 of 3

Page 75
SECTION - WATER
THE DETERMINATION OF CHLORINE DEMAND

Page 76

CHLORINE DEMAND
G-LAB METHOD No.21
(By K.P.Abraham)
1. SCOPE
This test method covers the determination of the chlorine demand in water samples, which does
not contain any nitrite or manganese compounds and ferric ion content should be less than
2 ppm.
2. PRINCIPLE
Chlorine demand is the quantity of chlorine that is reduced or converted to inert or less active
forms of chlorine by substances in the water. In most cases, chlorine demand implies complete
reaction with all chlorine reactable materials and is defined as the difference between the
amount of chlorine applied and the amount of free chlorine remaining at the end of the contact
period.
A standard chlorine solution is kept in contact with a known volume of the sample for 5
minutes. The residual chlorine is then estimated using potassium iodide and sodium
thiosulphate. The sodium thiosulphate is standardised against potassium iodate.
3. APPARATUS

3.2. Volumetric flasks, 1 litre, 500 ml
3.3. Pipettes, 2 ml, 10 ml
3.6. Measuring cylinders, 25 ml, 50 ml and 100 ml
3.7. Drying oven.
4. REAGENTS
4.1. Chlorine solution

50 ml of concentrated hydrochloric acid (HCl) is reacted with 5 g of potassium
permanganate (KMnO4) and the chlorine gas drawn off via a condenser to a water
trap. The resulting solution is diluted to 1 litre and stored in a refrigerator.
Alternatively, dilute household hypochlorite solution, which contains about 30,000 to
50,000 mg chlorine equivalent/L can be used.
4.2. Potassium iodate, standard (0.1M)

Dry about 5 g of potassium iodate (1(103) at 103 + 2°C for 1 hour. Cool in a
dessicator, and weigh 3.567 g, dissolve in distilled water and dilute to 1 litre. Store in
a glass stoppered bottle.
GLAB-21 (Revised) - Chlorine Demand Page 2 of 3

Page 77
4.3. Sodium thiosulphate, standard (0.1M)

Dissolve 25 g sodium thiosulphate (Na2S2O3.5H20) in freshly boiled distilled water
and make up to 1 litre.
4.4. Acetic acid, Concentrated

Glacial acetic acid.
4.5. Starch solution

Boil 500 ml distilled water. Make a cold water paste with 3 g of soluble starch. Add
this to the boiling water. Cool and store.
4.6. Potassium iodide (10%)
Dissolve 10 g potassium iodide (KI) in distilled water and make upto 100 ml.
5. STANDARDI ZATION
5.1. Standardization of thiosulphate solution
To 80 ml distilled water, add with constant stirring, 1 ml concentrated sulphuric acid,
10.00 ml standard potassium iodate solution (0.10M) and 1 g potassium iodide (KI).
Titrate immediately with the thiosulphate solution (0.lM) until the yellow colour of
the liberated iodine is almost discharged. Add 1 ml starch indicator solution and
continue titrating until the blue colour disappears.
Normality of Na2S2O3 = _______1___________

ml Na2S2O3 consumed
5.2. Standardization of chlorine solution

Place 2 ml acetic acid and 25 ml chlorine — demand — free water in a flask. Add
about 1 g Potassium iodide KI. Measure into the flask a suitable volume of chlorine
solution. Titrate with standardised thiosulphate solution (0.1M) until the yellow
iodide colour almost disappears. Add 1 to 2 ml starch indicator and continue titration
to disappearance of blue colour. Note the titre reading.
Chlorine, mg/l = Titre reading x Normality of thiosuluhate x 0.71

Volume of chlorine solution.
6. PROCEDURE
To 500 ml of the sample add 5.0 ml of the standard chlorine solution and mix. Allow to stand
for 5 minutes (exactly), then place 100 ml of the sample in a 250 ml conical flask add 5 ml
glacial acetic acid, 2 ml 10% potassium iodide (KI) and a few drops of starch solution. Titrate
with the standard thiosulphate solution until the blue starch iodide colour disappears. Note the
titre reading as V2.
Pipette 5.0 ml of the standard chlorine solution into a volumetric flask and dilute to 500 ml.
Titrate 100 ml of this solution as above. Let the titre be V1
7. CALCULATION
Chlorine demand, mg/l = (V2 — V1) x Normality of thiosulphate x 0.71.
Note: The chlorine demand increase with increase in contact time and also varies with the
dose rate and temperature.
GLAB-21 (Revised) - Chlorine Demand Page 3 of 3

Page 78
SECTION: WATER
THE DETERMINATION OF CHROMATE

Date issued : 23rdMarch 1995
Page 79
TEST METHOD FOR COOLING WATER

CHROMATE (COLOURIMETRIC)
G-LAB METHOD No.22
(By K.P.Abraham)
1. SCOPE
This test method is used for the determination of chromate in cooling water, which is dosed
with chromates, e.g. Gas turbine cooling water.
2. PRINCIPLE
Chromates when dissolved in water give an intense yellow colour. The intensity of this colour
is proportional to the concentration of chromate. The intensity of the colour is measured
photometrically at 440 nm, using distilled water as blank.
3. APPARATUS
3.1. Spectrophotometer — JASCO, Model V - 530

3.2. Cells 10 mm.
3.3. Drying oven
3.5. Volumetric flask, 1 litre and 100 ml, 6 Nos.
3.6. Pipettes, 10, 20 and 50 ml.
4. REAGENTS
4.1. Chromate solution, standard (1000 mg/l)
Dry potassium chromate AR at l50°C for 2 hours. Weigh 1.6741 g of the crystals and
make upto 11 in a volumetric flask using distilled water.
5. CALIBRATION
Prepare dilutions of 100 mg/l , 200 mg/l , 300 mg/l , 400 mg/l and 500 mg/l from the standard
chromate solution (1000 ppm), by pipetting 10, 20, 30, 40 and 50 ml to 100 ml volumetric flask
and making upto the mark with distilled water. Set the spectro photometer wave length at 440
nm. Take distilled water in 10 mm cell and adjust the transmittance and absorbance at infinity
and zero respectively. Take various dilution of the chromate prepared, in different cells and
measure the absorbance. Then draw a calibration graph with absorbance against concentrations.
Calculate the factor F’ from the graph, where ‘ F ’is mg/litre of chromate which produces unit
absorbance.
6. PROCEDURE
Take sample in a cell and using distilled water as blank measure the absorbance. If absorbance
is more than 1.0, give suitable dilution. If sample is turbid, filter through a fast filter paper
(No.41).
GLAB-22 (Revised) - Chromate (Colourimetric) Page 2 of 3

Page 80
7. CALCULATION
Chromate as Cr04 mg/l = Absorbance x F where F is the factor from calibration graph.
Chromate as MgCrO4, mg/litre = Chromate as Cr04 mg/l x 1.21
(Note: If sample is diluted, dilution to be accounted for in the calculation).
GLAB-22 (Revised) - Chromate (Colourimetric) Page 3 of 3

Page 81
G—STATION LABORATORY METHOD
SECTION: WATER
THE DETERMINATION OF FECAL COLIFORM BACTERIA

Date issued : 25thMarch 1995
Page 82

FECAL COLIFORM BACTERIA
G-LAB METHOD No.23
(By K.P.Abraham)
1. SCOPE
This method covers the determination of fecal coliform bacteria in drinking water, blended
water and treated waters. Fecal coliform analysis is a more definite test for recent fecal
pollution than the total coliform test, and fecal coliform is the standard test organism used in
many laboratories testing treated sewage, untreated public water supplies and such primary
contact waters as swimming areas.
2. PRINCIPLE
100 ml of water (collected in a sterile container) is filtered thorough a Millipore filter under
sterile conditions. The bacteria are retained on the filter. This filter is then placed on top of
MFC medium containing lactose, protein digest, vitamins, selected chemicals and Aniline blue
dye. The membrane is incubated for 24 hours at 44.5°C + 0.2, allowing coliforms of fecal
origin only to grow into visible colonies. Non-fecal colonies will not grow due to the heat. As
the fecal colonies grow they ferment lactose and the product acid reacts with the aniline dye to
produce a blue colour. Only colonies exhibiting a blue colour are fecal colonies and these are
counted.
3. APPARATUS
3.1. Bottles
Sampling bottles should be washed and sterilised with great care. Wrap the cap of the
bottle with aluminium foil. Dry heat at 1700C for 1 hour or autoclave at 121°C for 15
minutes at one bar or sterilise in a UV steriliser for five minutes.
3.2. Milliflex-100 vacuum pump.
3.3. Milliflex—100 Unit,(funnel with filter - sterile)
3.4. Milliflex liquid medium cassettes
3.5. Forceps
3.6. Alcohol burner
3.7. Autoclave KSG 113
3.8. Incubator at 44.5°C
3.9. Colony counter, Suntex 560
3.10. Sterile 47mm filter, 0.45 micron pore size.
4. REAGENTS
4.1. Sterile Phosphate Buffer Water

4.1.1. Dissolve 34.0 g of potassium dihydrogen phosphate in 500 ml of distilled
water.
GLAB-23 (Revised) - Fecal Coliform Bacteria Page 2 of 4
Page 83
4.1.2. Adjust the pH to 7.2 with 1N Sodium Hydroxide solution.

4.1.3. Dilute to 1 litre to produce stock buffer solution I, keep in refrigerator.
Discard if it becomes turbid.
4.1.4. Dissolve 50 g of magnesium sulphate (MgSO4.7H20) in distilled water and
dilute to 1 litre — to produce stock solution II.
4.1.5. Add together 1.25 ml of stock solution I and 5 ml of stock solution II and
dilute to 1 litre with distilled water to make the working solution.
4.1.6. Sterilise (autoclave at 1210C at 15 psi for 15 minutes, or UV
irradiate). Filtration through a sterile filter membrane to a
flask which is sterilised by heating in an oven at 1700C for 1
hour is also recommended.
4.1.7. Store in refrigerator.
4.2. m-FC Broth
From prepared sterile ampoule.
5. PROCEDURE
5.1. Sampling
5.1.1. After sterilisation and cooling add 0.1 ml of 10% sodium thiosulphate for
each 120 ml of bottle capacity. This removes residual chlorine from the
sample.
5.1.2. Ensure the sample point is not leaking.
5.1.3. Allow sample to run to waste.
5.1.4. Shut off flow sterilise the sample point with a flame from an alcohol
burner. Open valve and flush for a short time.
5.1.5. Fill sample bottle but leave an air space so that the sample may be shaken.
5.1.6. If not tested within one hour of sampling, the bottle should be kept in a ref
irgerator. Up to 30 hours storage is permissable.
5.2. Laboratory Procedure
5.2.1. Ensure equipment is completely sterilised before use (including the forceps
in the alcohol burner flame).
5.2.2. Prepare the liquid medium cassette as follows:
Remove the yellow cap from the cassette. Open the medium ampoule by
twisting off the ampoule top. Insert the Luer male top of the ampoule into
the Luer female connection of the cassette. Squeeze the ampoule to
dipense the medium into the cassette. Make sure to use the entire content
of the ampoule. Remove the empty ampoule and replace the yellow cap.
The protective cover remains on the cassette and is only removed prior to
using the cassette.
5.2.3. Sanitize the pump head with alcohol wetted gauze. Place the aseptic spacer
on the vacuum support of the pump. Place the incubation cassette on the
work surface.
5.2.4. Place a Milliflex filter funnel on the vacuum support of the pump, pushing
down until it seats correctlt.
5.2.5. Remove cover from the funnel. Shake the sample vigorously for several
seconds.Fill sample into the funnel upto the 100 ml mark. If you replace
the cover on the Milliflex, don’t close completely.
Page 84
5,2.6. Operate the vacuum pump to filter the sample. Wash the funnel with sterile
buffer — 2 washes of 30 ml. After pump has been stopped manually, wait
for red indicator light to switch off, indicating venting of vacuum system.
After the indicator light has switched off, remove funnel from vacuum
support and replace cover on the funnel.
5.2.7. Verify membrane integrity through visual inspection: the membrane has
adopted a convex shape.
5.2.8. Open incubator cassette and place funnel onto the cassette. The membrane
is now fully in contact with the medium.
5.2.9. With the palm of your hand apply a firm vertical pressure onto the funnel.
The funnel will shear off at the recess provided.
5.2.10. Remove the lid from the funnel and place it onto the cassettre to protect the
membrane. Dispose of the funnel. Incubate for 22 — 24 hours at 44.5°C +
0.2. The petri dish must be inverted to prevent droplets of condensation
spoiling the developing colonies.
5.2.11. After the required period remove from incubator, turn the dish correct way
up.
5.2.12. Examine the surface of filter for colonies using the colony counter.
Fecal colonies are recognised by their blue colour. Cream or grey colonies
are not fecal coliform. It is important, in order to avoid errors due to subtle
colour changes, that the filter is examined within 30 minutes of removal
from the incubator.
6. CALCULATION
No of blue colonies/100ml is reported as No. Fecal Coliform per 100 ml.
Reference = APHA 9222 D

International standards for drinking water WHO 1971
Coliform analysis — Millipore Corporation.

Page 85
SECTION: WATER
THE DETERMINATION OF TOTAL COLIFORM BACTERIA

Page 86

TOTAL COLIFORM BACTERIA
G -LAB METHOD No.24
(By K.P. Abraham)
1. SCOPE
This method covers the determination of total coliform bacteria in drinking water, blended
water and treated waters. Coliform organisms, while relatively harmless themselves, are almost
always present in water that contains enteric pathogens. Thus, because they are relatively easy
to isolate and because they normally survive longer than the disease-producing organisms,
coliforms are a useful indicator of the possible presence of enteric pathogenic bacteria and
viruses. In most cases water that is free of total coliforms is considered free of diseases
producing bacteria.
2. PRINCIPLE
100 ml of water (collected in a sterile container) is filtered through a millipore filter under
sterile conditions. The bacteria are retained on the filter. This filter is then placed on top of
MF—Endo media containing lactose, protein digest, vitamins, selected chemicals and Schiff’s
Reagent. The membrane is incubated at 35°C for 24 hours, the media diffuses through the
membrane and supplies nutrient to the bacteria. The coliform bacteria present will ferment the
lactose forming an acid-aldehyde complex which reacts with the Schiff’s Reagent to form an
irridescent green coating over the growing colonies. These colonies are counted.
3. APPARATUS
3.1. Bottles
Sampling bottles should be washed and sterilised with great care. Wrap the cap of the
bottle with aluminium foil. Dry heat at l70°C for 1 hour or autoclave at l21°C for 15
minutes at one bar or sterilise in a UV steriliser for five minutes.
3.2. Milliflex—100 vacuum pump.
3.3. Milliflex—l00 Unit,(funnel with filter —sterile)
3.4. Milliflex liquid medium cassettes
3.5. Forceps
3.6. Alcohol burner
3.7. Autoclave KSG 113
3.8. Incubator at 44.5°C
3.9. Colony counter, Suntex 560
3.10. Sterile 47mm filter, 0.45 micron pore size.
GLAB-24 (Revised) - Total Coliform Bacteria Page 2 of 5

Page 87
4. REAGENTS
4.1. Sterile Phosphate Buffer Water
4.1.1. Dissolve 34.0 g of potassium dihydrogen phosphate in 500 ml of distilled water.
4.1.2. Adjust the pH to 7.2 with lN Sodium Hydroxide solution.
4.1.3. Dilute to one litre to produce stock buffer solution I, keep in refrigerator. Discard
if it becomes turbid.
4.1.4. Dissolve 50 g of magnesium sulphate (MgSO4.7H20) in distilled water and dilute
to 1 litre to produce stock solution II.
4.1.5. Add 1.25 ml of stock solution I and 5.0 ml of stock solution II in a 1 litre flask and
make up with distilled water to make the working solution.
4.1.6. Sterilise (autoclave at 121°C at 15 Psi for 15 minutes, or UV irradiate). Filtration
through a sterile filter membrane to a flask which is sterilised by heating in an
oven at 170°C for 1 hour is also recommended.
4.1.7. Store in refrigerator.
4.2. m-FC Broth

Ampoule (2m1) M-ENDO BROTH, Cat. No. M000 000 2E by Millipore
Corporation.
5. PROCEDURE
5.1. Sampling
5.1.1. After sterilisation and cooling add 0.1 ml of 10% sodium thiosulphate for each
120 ml of bottle capacity. This removes residual chlorine from the sample.
5.1.2. Ensure the sample point is not leaking.
5.1.3. Allow sample to run to waste.
5.1.4. Shut off flow, sterilise the sample point with a flame from an alcohol burner.
Open sampling valve and flush for a short time.
5.1.5. Fill sample bottle but leave an air space so that the sample may be shaken.
5.1.6. If not tested within one hour of sampling, the bottle should be kept in a
refrigerator. Up to 30 hours storage is permissible.
5.2. Laboratory Procedure
5.2.1 Ensure equipment is completely sterilised before use (including the forceps in the
alcohol burner flame).

Page 88
5.2.2. Prepare the liquid medium cassette as follows:

Remove the yellow cap from the cassette. Open the medium ampoule by twisting
off the ampoule top. Insert the Luer male top of the ampoule into the Luer female
connection of the cassette. Squeeze the ampoule to dipense the medium into the
cassette. Make sure to use the entire content of the ampoule. Remove the empty
ampoule and replace the yellow cap. The protective cover remains on the cassette
and is only removed prior to using the cassette.
5.2.3. Sanitize the pump head with alcohol wetted gauze. Place the aseptic spacer on the
vacuum support of the pump. Place the incubation cassette on the work surface.
5.2.4. Place a Milliflex filter funnel on the vacuum support of the pump, pushing down
until it seats correctly.
5.2.5. Remove cover from the funnel. Shake the sample vigorously for several
seconds.Fill sample into the funnel up to the 100 ml mark. If you replace the
cover on the Milliflex, don’t close completely.
5.2.6. Operate the vacuum pump to filter the sample. Wash the funnel with sterile
buffer-2 washes of 30 ml. After pump has been stopped manually, wait for red
indicator light to switch off, indicating venting of vacuum system. After the
indicator light has switched off, remove funnel from vacuum support and replace
cover on the funnel.
5.2.7. Verify membrane integrity through visual inspection: the membrane has adopted
a convex shape.
5.2.8. Open incubator cassette and place funnel onto the cassette. The membrane is now
fully in contact with the medium.
5.2.9. With the palm of your hand apply a firm vertical pressure onto the funnel. The
funnel will shear off at the recess provided.
5.2.10. Remove the lid from the funnel and place it onto the cassette to protect the
membrane. Dispose of the funnel. Incubate for 22 - 24 hours at 35° C + 0.2.
The petri dish must be inverted to prevent droplets of condensation spoiling the
developing colonies.
5.2.11. After the required period remove from incubator, turn the dish correct way up.
5.2.12. Examine the surface of filter for colonies using the colony counter. Total coliform
colonies are recognised by their “green sheen”.
6. CALCULATION
No of green sheen colonies/l00ml is reported as No. Total Coliform per 100 ml.
Reference : APHA 4222 B

International Standards for drinking water - WHO 1971
Coliform analysis — Millipore Corporation.
m-ColiBlue24 Broth data sheet

Page 89

Page 90
DUBAI ELETRICITY AND WATER AUTHORITY
SECTION: WATER
THE DETERMINATION OF TOTAL COLIFORM BACTERIA

(MPN Method)

Page 91

TOTAL COLI FORM BACTERIA (MPN METHOD)
G-LAB METHOD NO. 25
(By K.P. Abraham)
1. SCOPE
Microbial examination of water samples, are done to determine the sanitary quality of water.
This method covers the determination of total coliform bacteria in drinking water, blended
water, treated waters, salty or brackish waters as well as mud, sediments and sludge. Coliform
organisms, while relatively harmless themselves, are always present in the faeces of man and
other warm blooded animals and hence in sewage, in large numbers. The presence of such fecal
indicator organism in a sample of drinking water thus denotes that intestinal pathogens could be
present, and that the supply is there fore potentially dangerous to health, Thus because they are
relatively easy to isolate and because they normally survive longer than the disease producing
organisms, coliforms are a useful indicator of the possible presence of enteric pathogenic
bacteria and viruses. In most cases water that is free of total coliform is considered free of
disease producing bacteria.
2. PRINCIPLE
Coliforms are a bacterial group of organisms, They may be aerobic or facultative anaerobic,
gram negative, non spore forming, rod-shaped. They ferment lactose forming acid and gas
within 48 hours at 35°C. The standard test method for coliform group may be multiple tube
fermentation technique (MPN) or membrane filter technique (MF). By MPN method the
sample of varying volumes are inoculated into an array of fermentation tubes. By MPN method
the precision of each test depends on the number of tubes used. The most satisfactory
information will be obtained when the largest sample inoculums examined shows gas in some
or all of the tubes and the smallest sample inoculums show no gas in all or a majority of the
tubes. Bacterial density can be estimated either by using the formula given or from the table
using the number of positive tubes in the multiple dilutions. MPN tables are based on their
assumption of a random dispersion of the coliform. So the sample bottles are to be shaken
thoroughly before taking inoculums from it. To obtain a more precise estimate of bacterial
density in treated drinking water, which should contain no coliforms per 100 ml, use ten
replicate tubes of 10 ml each or 100 ml sample portions. The MPN technique is applicable in
the analysis of salt or brackish water as well as mud, sediments and sludge. Solid or semisolid
samples are diluted ten times (50 g sample + 450 ml diluent) using sterile phosphate buffer or
0.1% peptone dilution water. Homogenize the sample with mixer and take decimal dilutions as
early as possible and inoculate the fermentation tubes. MPN technique involves three phases in
the analysis. 1) Presumptive test, 2) confirmed test and, 3) completed test. The fecal coliform
test differentiates between coliforms of fecal origin (intestines of warm blooded animals) and
coliforms from other sources.
3. APPARATUS
3.1. 10 ml fermentation tubes with inverted vials.

3,2. Incubating oven with minimum temperature precision of ± 0.5°C.
3.3. Steam sterilizer (auto clave).
3.4. Petri-dishes.
3.5. Platinum wire loop.
GLAB-25 (Revised) - Total Coliform (MPN method) Page 2 of 7

Page 92
3.6. Microscopic glass slides,

3.7. Glass slide covers,
3.8. Burners.
3.9. Slide stands,
3.10. Circulating water bath with minimum temperature precision of ±0.2 C.
4. REAGENTS
4.1. Lactose broth

Dissolve 13 grants of dehydrated lactose broth into 100 ml demin water adjust pH to 6.9
±0.2. Before sterilizing dispense sufficient medium in fermentation tubes (1 ml) with an
inverted tube inside the vials,
4.2. Brilliant green lactose bile broth
Add 40 grams in one litre demin water and heat to dissolve. pH should be 7.2 ±0.2 after
sterilization,
4.3. Peptone water
Dissolve 10 grams in 100 ml, adjust pH to 6.8 and autoclave for 15 minutes at 121°C.
4.4. Nutrient agar
Take 23 grams of ready-made supply and dissolve in one litre of demin water.
4.5. Gram stain reagents
4.5.1. Ammonium oxalate — crystal violet, (Hackers’)
Dissolve 2 grams of crystal violet (90% dye content) in 20 ml 95% ethyl alcohol;
dissolve 0,8 grams of ammonium oxalate (NH4)2C2O4,H2O in 80 ml demin water
mix the two solution and store 24 hours before use, Filter through paper into
staining bottle
4.5.2. Lugol’s solution, (gram’s modification)
Grind 1 gram iodine crystals and 2 grams of KI in a mortar, add a little demin
water to dissolve completely while grinding. Dilute to 300 ml and store in an
amber glass bottle,
45.3. Counter stain
Dissolve 2.5 grants of safranin dye in 100 ml 95% ethyl alcohol, Add 10 ml of
this to 100 ml demin water.
4.5.4. Acetone alcohol mixture
Mix equal volumes of ethyl alcohol 95% with acetone.
4.6. EC medium
Take 37 grams and dissolve in one litre demin water by heating, pH should be 6.9 ±0.2
after sterilization, Transfer sufficient media into the fermentation tubes (1 ml) with an
inverted tube inside the vials,
5. PROCEDURE (for drinking water)
5.1. Presumptive test

5.1.1. Arrange 5 ten ml vials and dispense 1 ml of concentrated media into it and
sterilize for 15 minutes at 121°C and cool,
5.1.2. Add 10 ml of representative sample into each vials shake well and remove air
bubbles from the inverted tubes. For non potable polluted waters use 5 tubes
per dilution (of 10, 1 and 0.1 ml etc) and make up the vial volume using
peptone water.
Page 93
5.1.3. Incubate inoculated tubes at 35 ±0.5°C. After 24 hours examine tubes for gas.
If no gas produced inside the vials, incubate for 24 hours more. At the end,
examine the tubes again. The absence of gas or acid at the end constitutes
negative tests and the presence of gas and acid shows a positive presumptive
reaction. Submit the positive presumptive reaction to the confirmed phase.
5.2. Confirmed test

Schematic outline of confirmed phase,
Inoculate lactose broth fermentation tubes and incubate 24 ±2h at 35 ±0.5°C
(1) (2)
Gas or acidity produced, No gas or acidic growth,

Transfer to incubate additional
Brilliant green lactose 24 h (total 48 + 3 h )
bile broth, Incubate
48 ±3h at 35 ±0.5°C
(a) (b) (a) (b) (c)
Gas No Gas Gas Acidic No gas or

Produced. Produced Produced growth, acidic
Coliform Negative confirm confirm growth
Group test. as in (l ) as in ( l ) produced.
Confirmed Coliform Negative
Group test.
absent coliform
group absent
5.2.1. As soon as gas formation takes place in the presumptive tubes transfer to the
confirmatory medium. Do not have to wait to complete the 48 hours duration of
incubation, For transferring organism from one media to other just change the
caps after shaking the tubes,
5.2.2. Incubate the inoculated brilliant green lactose bile broth tube for 48 hours at 35
±0.05°C, Formation of gas in any amount in the inverted vial of the BGL broth
fermentation tubes at any time within 48 ±3 hours constitutes a positive
confirmed phase,
5.2.3. Calculate the MPN value from the number of positive brilliant green lactose bile
tubes. If all the presumptive tubes are positive in two or more consecutive
dilutions within 24 hours, submit to the confirmed phase only the tubes of
highest dilution (smallest sample inoculum) in which all tubes are positive and
any positive tubes in still higher dilutions, Submit to the confirmed phase all the
tubes in which gas or acidic growth is produced only after 48 hours,
5.3. Completed test
5.3.1. To establish definitively the presence of coliform bacteria and to provide quality
control data, use the completed test on all the positive confirmed tubes, double
Page 94
confirmation into brilliant green lactose bile broth for total coliforms and EC
broth for fecal coliforms.
5,3.2. Using aseptic technique streak one LES endo agar (alternately macConkey agar
also may be used) plate from each tube of BGL bile broth showing gas. Incubate
plates at 35 ±.5°C for 24 ±2 hours.
5.3.3. Typical colonies developed (pink to dark red with a green metallic sheen) on LES
endo agar/MacConkey agar are transferred into a nutrient agar slant and into a
single strength lactose broth vial, Incubate the secondary lactose broth vials at 35
±0.5°C for 24 or 48 hours.Microscopically examine the gram stained preparation
from those 24 hour nutrient agar slant culture.
5.3.4. Gram stain technique: (the gram stain nay be omitted from the potable water
samples).
5.3.4.1 Prepare a light emulsion of the test bacterial growth using demin water.
5.3.4.2 Transfer a few drops of the emulsion into the slide.
5.3.4.3 Air dry and fix by passing the slide through a flame,
5.3.4.4 Stain with ammonium oxalate—crystal violet solution for 1 minute.
5.3.4.5 Rinse the slide in tap water and drain off excess.
5.3.4.6 Apply lugols’ solution for 1 minute and rinse the stained slide in tap water.
5.3.4.7 Decolorize for approximately 15 to 30 seconds with acetone alcohol
mixture by holding slide between fingers and letting the mixture flow
across the stained smear until the solvent flows colourlessly.
5.3.4.8 Counter stain with safranin for 15 seconds,
5.3.4.9 Rinse with tap water, blot dry and examine microscopically. Gram-
positive organisms are blue and gram-negative organisms are red.
Interpretation
Formation of gas in the secondary tube of lactose broth within 48 ±3 hours and
demonstration of gram negative nonspore forming rod shaped bacteria from the agar culture
constitute a positive result for the completed test, dennnst rating the presence of a netter of
the coliform group.
5.4. Fecal coliform
5.4.1. The fecal coliform test differentiates between coliforms of fecal origin (intestines of
warm blooded animals) and coliforms from other sources.
5.4.2. Submit all the presumptive fermentation tubes showing any amount of gas or heavy
growth within 48 hours of incubation to the confirmed test.
5.4.3. Gently shake or rotate presumptive fermentation tubes showing gas or heavy
growth, With a sterile 3 mm diameter metal loop or sterile wooden applicator stick
transfer growth from each presumptive fermentation tube to EC broth.
5.4.4. Incubate inoculated EC broth tubes in a water bath at 44.5 ±0.2 °C for 24 ±2 hours.
Place all EC tubes in water bath within 30 minutes after incubation, Maintain
sufficient ‘hater depth in water bath incubator to immerse tubes to upper level of the
medium.
5.4.5. Gas production in an EC broth culture within 24 hours or less is considered a
positive fetal coliform reaction. (no gas or negative test at EC medium and presence
of gas or positive test at BGL bile broth indicate presence of non fecal coliforms).

Page 95
6. CALCULATION
6.1 Total nunher of coliform

Per 100 ml (by MPN table*) = Read the values from the chart corresponding
the combinations of positive coliform results
from the BGLB broth fermentation tubes.
6.2. Total number of fecal coliforms = Read the values from the table
per 100 ml (by MPN table*) corresponding to the combinations
of positive fecal coliform
results of the EC medium.
* MPN table.
While the MPN tables and calculations are described for use in the coliform test) they are
equally applicable in determining the MPN of any other organisms provided suitable test media
are available.
The MPN for combinations not appearing in the under given tables, or for other combinations
of tubes or dilutions, nay be estimated by Thomas’ simple formula as follows:
MPN /100 ml = ___________no of positive tubes X 10____________________

__________________________________________________
√ (ml sample in negative tubes X ml sample in all tubes)
MPN index value MPN index value

(when five 10 ml portions are used) (when ten 10 ml portions are used)
Number of tubes
Number of giving positive
MPN/100ml
tubes giving reaction out of 10 of
positive 10 ml each
reaction out MPN/100 ml <1.1
0
of 5 of 10 ml
each 1 1.1
0 <2.2 2 2.2
3 3.6
1 2.2
4 5.1
2 5.1
5 6.9
3 9.2
6 9.2
4 16.0 7 12.0
5 >16.0 8 16.1
9 23.0
10 >23.0

Page 96
MPN index for, various combinations of positive results, when sets of five tubes are used for
each sample portions of 10 ml, 1.0 ml and 0.1 ml (all diluted to 10 ml).
Combination MPN index / Combination MPN index / Combination MPN index

of positives 100ml of positives 100ml of positives / 100ml
0-0-0 <2 3-1-1 14 5-2-0 50

0-0-1 2 3-2-1 17 5-2-1 70
0-1-0 2 4-0-0 13 5-2-2 90
0-2-0 4 4-0-1 17 5-3-0 80
1-0-0 2 4-1-0 17 5-3-1 110
1-0-1 4 4-1-1 21 5-3-2 140
1-1-0 4 4-1-2 26 5-3-3 170
1-1-1 6 4-2-0 22 5-4-0 130
1-2-0 6 4-2-1 26 5-4-1 170
2-0-0 4 4-3-0 27 5-4-2 220
2-0-1 7 4-3-1 33 5-4-3 280
2-1-0 7 4-4-0 34 5-4-4 350
2-1-1 9 5-0-0 23 5-5-0 240
2-2-0 9 5-0-1 30 5-5-1 300
2-3-0 12 5-0-2 40 5-5-2 500
3-0-0 8 5-1-0 30 5-5-3 900
3-0-1 11 5-1-1 50 5-5-4 1600
3-1-0 11 5-1-2 60 5-5-5 >1600
When more than three dilutions are used in a decimal series of dilutions, use the results from the
only three of these in computing the MPN. To select the three dilutions to be used in determining
the MPN index, choose the highest dilution that gives positive results in all most of the five
portions
tested and the two next succeeding higher dilutions. Use the results of these three volumes in
computing the MPN index.
If the combinations of 10, 1.0 and 0.1 ml are used the MPN value is same as in the table.
If the combinations are the next dilutions i.e 1.0, 0.1 and 0.01 ml, then the
MPN value = Table value x 10.
If the contamination of the next dilutions i,e 0.1, 0.01 and 0.001 ml are used, then the
MPN value = Table value x 100
References: 1) APHA 9221, 1989,

2) Bacteriological examination of drinking water supplies 1982, Department of
environment, London.

Page 97
SECTION: WATER
THE DETERMINATION OF CONDUCTIVITY OF WATER

Date revised : 02nd June 2008
Page 98

CONDUCTIVITY
G-LAB METHOD No.26
(By K.P. Abraham)
1. SCOPE
This method covers the determination of conductivity of water samples i.e.; drinking water,
deionised water, cooling water, boiler water etc. Conductivity measurements are used for
1.1 Establish degree of mineralization to assess the effect of the total concentration of
ions on chemical equilibria, physiological effect on plants or animals, corrosion rate
etc.
1.2 Assess degree of mineralization of distilled water and deionised water.
1.3 Evaluate variations in dissolved mineral concentrations of raw water or wastewater.
1.4 Estimate sample size to be used for common chemical determinations.
1.5 Determine amount of ionic reagent needed in certain precipitation and neutralization
reactions.
1.6 Estimate total dissolved solids in a sample by multiplying conductivity by an

empirical factor.
2. PRINCIPLE
Conductivity is a numerical expression of the ability of an aqueous solution to carry an electric
current. It is measured as the reciprocal of the resistance in ohms measured between the
opposite faces of a centimetre cube of an aqueous solution at a specified temperature.
3. APPARATUS
3.1 Digital conductivity meter
Microprocessor conductivity meter, WTW
Model LF 537
3.2 Thermometer-capable of being read to the
nearest 0.1 °C and covering 0-100°C.
3.3 Conductivity cell-Electrode type with known cell

constant. WTW cell, Model LTA 01 or 1
3.4 Automatic Temperature Compensation probe. WTW

temperature sensor, Model TFK 530
4. REAGENTS
4.1 Conductivity water — distilled water with conductivity less than 1 µmho/cm.
4.2 Standard potassium chloride solution (KC1 0.0100M).
Dissolve 745.6 mg anhydrous KC1 in the conductivity water and dilute to 1000 ml at
GLAB-26 (Revised) - Conductivity Page 2 of 3

Page 99
25°C. This is a standard reference solution, which at 25°C has a conductivity of 1413
µmhos/cm. A second reference solution for lower conductivity can be made by
diluting the above standard by 10 times using the conductivity water i.e. 0.0010 M
KC1 at 25°C has a conductivity of 147 µmhos/cm.
5. CALIBRATION
Slide the reference temperature switch at the back side of the instrument to 25°C. Press the °C
push button until function C is selected. Press Up or Down push button until the display
indicates the cell constant of the cell which is being used. Press the °C push button until
function TC is selected, Press Up or Down push button until the display indicates 0.00. Rinse
the conductivity cell with distilled water. Then rinse the cell with standard KC1 solution
(0.0100 M). Take the standard KC1 solution in a beaker and immerse the electrode and ATC
probe in it. Note the conductivity reading displayed in the meter. If it is different from the
standard value of 1413 µmhos/cm, press the °C push button until function C is selected. Press
Up or Down push button until the conductivity display indicate the value 1413 µS/cm. Press
the °C push button until function TC is selected. Press Up push button until the display
indicates nLF. Press the °C push button again to select function °C.
6. PROCEDURE
Rinse the electrode with distilled water and then with the sample. Then take the sample in a
beaker and immerse the electrode and ATC probe and leave it for few minutes to stabilise.
Take the reading.
7. CALCULATION
Report the meter reading as micro mhos per centimetre (µmhos/cm) or microsiemens per
centimetre (µS/cm) at 25°C.
In the international system of units (SI) the conductivity is reported as millisiemens per metre
(mS/M)
1 mS/M = 10 µmhos/cm
1 mS/cm = 1000 µS/cm

ASTM D 1125
GLAB-26 (Revised) - Conductivity Page 3 of 3

Page 100
SECTION: WATER
THE DETERMINATION OF COPPER (LOW RANGE)

Page 101

COPPER (LOW RANGE)
G-LAB METHOD No.27
(By K.P. Abraham)
1. SCOPE
This method is applicable to the determination of copper in water such as steam condensate,
distilled water as well as in sea water and recirculated brine. It is specifically applicable to
concentrations of copper from 2 ~100 µg/litre (ppb).
2. PRINCIPLE
The method is based on the measurement of intensity of the blue colour of the cuprous complex
of zincon. The intensity of the colour is proportional to the copper concentration. Measurement
is done at 600 nm using 50 mm cells.
3. APPARATUS
3.1. Spectrophotometer
JASCO V 530.
3.2. Sample cells, 50 mm.
3.3. Erlenmeyer conical flasks, 250 ml (10 Nos.)
3.4. Pipettes, 1, 2, 4, 5, 10 & 20 ml.
3.5. Measuring cylinder - 250 ml.
3.6. Volumetric flasks, 1 litre, 200 ml.
3.7. Water polishing unit, Millipore – Milli-Q - gradient
4. REAGENTS
4.1 De-ionized water (Copper free).

Because most ordinary distilled water contains detectable amounts of copper, use de-
ionized water, prepared by passing distilled water through a water polishing unit, to
prepare all reagents and for dilutions.
Water polishing unit Millipore Milli-Q gradient is found suitable for this purpose.
GLAB-27 (Revised) - Copper (Low range) Page 2 of 4

Page 102
4.2 Stock copper solution, standard (1 ml = 1 mg Cu).

Standard (1 ml = 1 mg Cu) as purchased.
(or)
Dissolve 1.000 ± 0.0005 g of pure copper (Rod, foil or wire) in 20 ml ± 0.5 ml of

nitric acid (1:1 dilution), contained in a covered 250 ml glass beaker, by warming
gently on a hot plate. When dissolved add 16 ml ± 0.5 ml of approximately 6N
hydrochloric acid and 100 ml water. Cool and dilute to 1litre in a volumetric flask.
Store in a glass stoppered bottle.
4.3 Copper solution, Standard (1 ml = 0.01 mg Cu)
Pipette 10 ml of the standard copper solution (1 ml = 1 mg Cu) to a 1 litre volumetric
flask and dilute to 1 L using copper free distilled water.
4.4 Copper solution, Standard (1 ml = 2 µg Cu)

Pipette 20 ml of the standard copper solution (1 ml = 0.01 mg Cu) to a 100 ml
volumetric flask and make up using copper free distilled water.
4.5 Tartaric acid solution (15 %)

Dissolve 150 g tartaric acid and make up to 1 L with DM water.
4.6 Hydrochloric acid Concentrated HC1 (Sp.gr. 1.19)
4.7 Ammonium acetate buffer solution (50 %)

Dissolve 500 g ammonium acetate (CH3COONH4) in DM water and make up to 1 L.
4.8 Zincon stock solution

Dissolve 0.075 g of zincon (C20H15SN4O6Na) in 50 ml methyl/ethyl alcohol. Dissolve
completely by warming and dilute to 100 ml with DM water. This should be stored in
a brown coloured bottle.
4.9 Zincon solution (working)

Dilute 10 ml of zincon stock solution to 100 ml with ethyl alcohol.
5. CALIBRATION
Pipette 1.0, 2.0, 4.0, 6.0, 8.0 and 10 ml of the standard copper solution (1 ml = 2 µg Cu) to a
200 ml volumetric flask and add remaining volume of copper free distilled water to make the
volume as 200 ml. (This will give 10,20, 40, 60, 80 and 100 µg/l Cu). Transfer these standards
to 250 ml erlenmeyer flasks. Take 200 ml of copper free distilled water as blank. Add 8.5ml of
concentrated hydrochloric acid to each and boil to approx. 50 ml and then cool. Add the
following solutions in the given order with mixing.
a) 25 ml of ammonium acetate solution
b) 2 ml of tartaric acid solution
c) 2 ml of zincon solution (working)
Dilute to 100 ml with DM water. After 10 minutes measure the absorbance at 600 nm
wavelength using 50 mm cell, against the blank. Plot a graph with absorbance versus
concentration (µg/l Cu). Calculate the factor ‘F’ from the graph, where F is µg/l Cu per unit
absorbance.

Page 103
6. PROCEDURE
Take 200 ml sample each in 250 ml erlenmeyer flasks. Take 200 ml of copper free distilled
water as blank. Add 8.5 ml of concentrated hydrochloric acid to each and boil to approx. 50 ml
and then cool. Add the following solutions in the given order with mixing.
a) 25 ml of ammonium acetate solution
b) 2 ml of tartaric acid solution
c) 2 ml of zincon solution (working)
Dilute to 100 ml with DM water. After 10 minutes measure the absorbance at 600 nm
wavelength using 50 mm cell, against the blank. Note the sample absorbance. In G -
Station laboratory, by calling method No.6 in JASCO spectrophotometer, all the
parameters required for the copper determination will be set in the instrument. The
concentration can be directly measured.
7. CALCULATION
Copper, µg/l Cu = Sample absorbance x F
Where ‘F’ is the factor from the calibration graph.
Reference: JIS K 01 01
JIS B 8224

Page 104
SECTION: WATER
THE DETERMINATION OF CORROSION RATE

(USING COUPONS)

Page 105

CORROSION RATE MONITORING USING COUPONS
G-LAB METHOD No.28
(By K.P. Abraham)
1. SCOPE
This method covers the determination of the corrosivity of water by evaluating pitting and by
measuring the weight loss of metal specimens. Pitting is a form of localized corrosion, weight
loss is a measure of the average corrosion rate. This method is applicable mainly to cooling
water.
2. PRINCIPLE
The rate of corrosion of a metal immersed in water is a function of the tendency for the metal
to corrode and is also a function of the tendency for the water and the materials it contains to
promote (or inhibit) corrosion.
Carefully prepared, weighed metal coupons are installed in contact with flowing cooling water
for a measured length of time. After removal from the system, these coupons are examined,
cleaned and reweighed. The corrosivity and fouling characteristics of the water are determined
from the difference in weight, the depth and distribution of pits, and the weight and
characteristics of the foreign matter on the coupon.
3. APPARATUS
3.1. Corrosion coupons

Approximately 3”x 3/8” of mild steel or material of the circuit.
3.2. Plastic holders and bolts for mounting
3.3. Plastic sample pipe work in a bypass circuit
GLAB-28 (Revised) - Corrosion rate (Coupon method) Page 2 of 4

Page 106
4. REAGENTS
4.1. Inhibited Hydrochloric acid

Mix 357 ml of concentrated HC1 (sp.gr 1.19) and 5.0 g of acid inhibitor. Then dilute
to 1 L with water.
4.2. Acetone
4.3. Acid Inhibitor.
5. PROCEDURE
5.1. Preparation of coupon
Take the corrosion coupon and remove any oil or grease on it using a solvent. Dry
and then sand blast to remove any oxides and then immerse it in the inhibited acid for
30 minutes. Remove the coupon from acid and wash thoroughly with distilled water.
Then dip the coupon in acetone. Dry and keep in the desiccators.
5.2. Measurement
Remove the coupon from desiccators and weight it. Install the coupon in the system
as shown in the diagram Fig. 1, using plastic nuts and bolts. Start the sample flow at
average 1 in/second ensuring coupon is totally immersed. After a period of not less
than 30 days remove coupon from the water stream dry and examine. Scrape oxides
or deposites from the coupon with a plastic implement. Wash with water and degrease
with acetone and dry. Clean with the inhibited hydrochloric acid. Rinse thoroughly
with distilled water, wash with acetone and dry in an oven at 105°C.Cool in a
desiccators and weigh. Calculate the weight loss (mg).
6. CALCULATION
6.1. Corrosion rate as penetration, mils per year
Penetration, mpy = Weight loss in mg x 143.7

No.of days x A x d
Where, mpy = mils per year

A = area of the coupon exposed in cm2
d = density of the metal in g/cm3
143.7 = 365
2.54
(1 year = 365 days, 1 inch = 2.54 cm, 1 mils = __1 __ inch )
1000
Density of metals : Mild steel = 7.83 g/cm3
Copper = 8.9 g/cm3
Brass = 8.70 g/cm3
Note:
When taking the area of the coupon, area of both sides of the coupon to be
considered and the area of the hole provided for fixing the coupon to be subtracted. If the
thickness of the coupon is considerable, the surface area of the sides also to be considered.

Page 107
6.2. Corrosion rate as weight loss, miligrams/sq.decimeters/day.
Weight loss, mdd = Weight loss in mg_______________________

Surface area of the coupon, din2 x No. of days
1 sq.decimeter = 100 sq.centimeters

l mpy = ~ 5mdd
Note: Duration for the test time can be approximately calculated using the
following equation.
T = 50_
CR
Where,
T = Time in hours
CR = Corrosion rate in millimeters/year
7. INTERPRETATION AND REPORTING
From the examination note the type of corrosion, See - Fig. 2
7.1. General uniform metal corrosion
7.2. Localized attack - large isolated areas of corrosion
7.3. Pitting attack - small isolated areas of corrosion.
7.4. Interpretation for 65—1000F.

0 - 2 mpy( 0 - 10mdd) = excellent protection
2 - 5 inpy(10 - 25mdd) = highly satisfactory
5 -10 mpy(25 - 50mdd) = tolerable
>10 mpy ( > 50 mdd) = Unsatisfactory

The NALCO Water Handbook.

Page 108
G—STATION LABORATORY METHOD
SECTION: WATER
THE DETERMINATION OF CORROSION RATE
(USING PROBES)

Page 109

CORROSION RATE MONITORING USING PROBES
G-LAB METHOD No.29
(By K.P. Abraham)
1. SCOPE
This method covers the determination of the corrosivity of water using a CORRATER system.
The CORRATER instrument can measure the general corrosion rate as well as the pitting
corrosion. The rate of corrosion of a metal immersed in water is a function of the tendency for
the metal to corrode and is also a function of the tendency for water and the materials it
contains to promote (or inhibit) corrosion. Since the two tendencies are inseparable, the
corrosiveness of a material or the corrosivity of water must be determined in relative, rather
than absolute terms.
With the use of a two-electrode probe, water having corrosion rate times resistivity product of
10,000 or less up to 25,000 with correction curve can be measured.
2. PRINCIPLE
CORRATER system operates on the fundamental principle that a metal corroding through
oxidation will generate a current. This method of determining corrosion is generally referred to
as Linear Polarization Resistance (LPR). A CORRATER instrument determines the corrosion
rate by measuring the current from a small applied potential difference between two or three
electrodes. Within certain limitations, this current can be related to the general corrosion rate.
In addition, CORRATER system can also measure pitting corrosion tendency. The pitting
index is actually the electrical current passing between the two electrode when the two
electrodes are connected to a zero—impedance ammeter.
The polarization resistance method of measuring corrosion rate comprises a means to measure
the electrical (electron-ion interchange) resistance at the interface between a metal element of
known surface area and a liquid without significantly disturbing the natural conditions of that
interface. The metal element, or the test element’, which is inserted into the liquid must closely
match the material of construction (pipe or vessel) for which the corrosion rate information is
required. The measured interfacial resistance (polarization resistance) is an inverse function of
the corrosion rate.
3. APPARATUS
3.1. Corrosion monitor

CORRATER Model RCS 9000 by
ROHRBACK COSASCO Systems.
3.2. CORRATER probe

Two electrode standard probe, Part
No.060800
3.3. Electrode elements

1010 Carbon steel, Part No.060814-8001
3.4. Standard test probe Part No.011000-1
GLAB-29 (Revised) - Corrosion rate (Probe method) Page 2 of 6

Page 110
4. REAGENTS
4.1. Acetone
As purchased.
4.2. Emery paper
For cleaning the electrodes.
5. CALIBRATION
The RCS 9000 instrument is calibrated to read corrosion rate in units of mils per year for
carbon steel and the common grades of stainless steels. For other alloys, a multiplier is required
on the instrument read out.
For verifying the instrument operation, connect the standard test probe and read it several times
on various functions. First, turn the unit ON, then select the 2EL STD CORR function. After
the delay time, a reading of 5 ±0.5 will appear. Next press 3EL STD CORR. The reading will
be virtually identical to the first one. This will generally be true for any low resistance
solutions. Now press 2EL FLUSH CORR. The reading will be about ten times larger than the
previous ones. Next, press 5T0 PIT. The reading should be less than 1.0.
6. PROCEDURE
6.1. Preparation of electrode

The electrode should have a clean and polished surface. If the electrode become fouled with
corrosion products or other materials, they should be cleaned and polished to a dull shine with
an emery cloth. After cleaning, the electrode should be thoroughly degreased with acetone and
handled with a clean cloth or paper towel to prevent contamination. Pre treatment of electrode
is done in order to stabilize the instrument readings. Generally, a full strength sample of the
treatment chemical is used. The new electrodes are carefully placed into the solution for a 6—
12 hour period and then threaded into the probe and placed into service.
CORRATER electrodes, when new, at 3/16”(4.76 mm) diameter by 15/12” (31.75 mm) long
cylinders. As corrosion occurs on the electrodes, their diameter decreases and at some point the
reduced diameter begins to affect the corrosion and pitting readings. It is recommended that
CORRATER electrode be replaced when their diameter has been reduced to 5/32” (3.97 mm).
6.2. Probe installation

The system shut down is required before the installation or removal of the probe. The
CORRATER probe must be carefully mounted where suspecting corrosion is occurring, not a
side stream, in order to obtain readings which represent the true corrosion experienced in the
system. The probe should not be positioned in such a way that one electrode shields the other
from full flow. Both the electrodes must always contact the flow to produce reliable corrosion
readings. To monitor side wall corrosion, length and placement should be chosen to be close to
the wall (Fig 1). When observing general corrosion, the probe should be projected into the
stream so that electrodes contact the center of the moving fluid (Fig 2).
Flow
Flow
Fig 1 Fig 2

Page 111
6.3. Measurement
6.3.1. To verify the instrument is functioning properly, attach the standard test
probe to the coiled cable, and follow steps in 6.3.2 and 6.3.3 below.
The display should read as follows:
Selection Reading Typical reading
2 or 3 EL CORR STD 5 ± 0.5 4.9

2 or 3 EL CORR FLUSH 50 ±5 49.5
PIT STD <1 0.1
PIT FLUSH < 10 1.2
6.3.2. Connect the RCS 9000 coiled cable to the probe which is installed in the
sample flow.
6.3.3. Press the ON key. The number 188.8 and LO BAT will appear momentarily.
The display will then change to 0.0 If the LO BAT sign stays on, this means
the battery is low and it needs to be replaced.
6.3.4. Select and press a function key. The unit will display the function selected
and then flash 00 while taking the reading. When the measure cycle is
complete (approx. 50 seconds), the reading will be displayed. The unit will
turn itself OFF if a new function is not pressed (approx. 2 minutes). Press
OFF to switch off the instrument after measurements.
Note: If a function switch is pressed during an actual reading, the current
measurement cycle will be aborted, and new cycle will begin.
7. CALCULATION
The instrument is calibrated for carbon and stainless steels. For other alloys, a multiplier (AM)
from Table 1 should be used.
Another correction factor (RC) to be used from the solution resistivity correction curve. The
factor (RC) to be read from Graph 1, corresponding to the instrument reading
(m.p.y.)/Conductivity (µmhos/cm).
Corrected corrosion rate = Instrument reading x AM x RC
Where,
AM = Alloy multiplier factor
RC = Resistivity correction factor
Reference: ASTM 2688

Manual for CORRATER RCS 9000

Page 112
TABLE 1
CORRATER MULTIPLIER FACTORS
NOTE: These factors are recommended for use with the Model 9000. They are based upon use of
CORRATER electrodes which have surface areas of 5cm2 for “standard” probes and 0.5 cm2 “flush”
probes.
UNS RCS Alloy

Material Multiplier
Code Code
K03005 8001-8008 Pipe Grade Carbon Steel 1.00
8009 BS 970 EM 38 1.00
S30400 8013 AISI 304 Stainless steel 0.97
S31600 8020 AISI 316 Stainless steel 1.00
S31603 8021 AISI 316 L Stainless steel 1.00
N08020 8043 Carpenter 20 CB3 SST 0.98
N04400 8054 Monel 400 Nickel 1.13
N05500 8055 Monel K-500 Nickel 1.04
N06600 8057 Inconel 600 Nickel 0.95
C71500 8060 CDA 715 70/30 Copper/Nickel 1.50
C11000 8061 Copper 110 ETP Comm. Pure 2.00
C70610 8062 CDA 706 50/10 Copper/Nickel 1.80
C68700 8064 CDA 687 Aluminium Brass Arsenical 1.62
C64200 8067 CDA 642 A1 Silicone Bronze 1.48
8068 CDA 442 Admiralty Uninhibited 1.67
C44500 8072 CDA 445 Phosphorized Admiralty 1.68
C44300 8073 CDA 443 Arsenical Admiralty 1.67
8075 Duronze IV 1.56
A91100 8080 Aluminium 1100-0 0.94
A92024 8085 Aluminium 2024 0.88
Z17001 8090 Grades 1A,1,2,3,4or 5 Zinc 1.29
R50400 8093 ASTMB-348 Grades 2-4 Titanium 0.75

Page 113

Page 114

SECTION: WATER
THE DETERMINATION OF DISSOLVED OXYGEN
(INDIGO CARMINE METHOD)

Page 115

DISSOLVED OXYGEN (COLOURIMETRIC INDIGO CARMINE)
G-LAB METHOD No.30
(By K.P. Abraham)
1. SCOPE
This method is applicable to water containing less than 60µg/l of dissolved oxygen, such as
steam condensate and deaerated boiler feed water only.
2. PRINCIPLE
Dissolved oxygen reacts under alkaline conditions with the indigo carmine solution to produce
a progressive colour change form yellow-green through red to blue and blue-green. The result
of each test can be determined by comparison of colour developed in the sample with colour
standard made up to represent different concentrations of dissolved oxygen.
3. APPARATUS
3.1. Lovibond Nessleriser Type AF 300 Mk 111
3.2. Special Nessler cylinder with vial for oxygen

measurement. Type AF 315
3.3. Standard lovibond Nessleriser disc NOE
3.4. Beaker, 2L
3.5. Small filler, Type AF 278.
3.6. Thermometer.
4. REAGENTS
Indigo carmine stock solution

Dissolve 1 indigo carmine/glucose tablet (BDH) in 5 ml of distilled water. Make up to 80 ml
with glycerol. Keep in a cool, dark place. Stable for 1 month Or it can be prepared by mixing
the following together:
Indigo carmine - 0.018 g
Glucose - 0.20 g
Distilled water - 5 ml
Glycerol - 75 ml
4.1. Potassium hydroxide solution (KOH 60%)

Dissolve 53 g potassium hydroxide (KOH) in distilled water and make up to 100 ml.
4.2. Leuco Reagent

Mix indigo carmine and KOH solution in the ratio of 4:1. Mix well. Leave until
colour changes to yellow. This should be made on the same day.
GLAB-30 (Revised) - Dissolved Oxygen (Indigo carmine method) Page 2 of 4

Page 116
5. PROCEDURE
Fill the inner glass vial of the Nessler cylinder to the top with leuco reagent using the filler.
With the glass tube provided drop the glass ball onto the surface of the leuco reagent. Ensure
no air bubbles are trapped. Fill a 2 litre beaker with cool sample (temp <2l°C). Connect a
polythene tube to the sample point. Insert the sample tube into the Nessler cylinder (with the
leuco reagent) and pass the sample (temp <21°C) and allow to overflow. Immerse the Nesslers
cylinder with sample flowing into the 2 litre beaker. Allow at least 5 minutes for the sample to
overflow. Keep the stopper under water. Remove sample line from the Nessler cylinder
carefully, without disturbing the glass ball on the top of the leuco reagent. Make sure all the
excess reagent is washed out from the cylinder. Replace the stopper. Invert the Nessler
cylinder, which is under water, to allow the glass ball to fall and the leuco reagent to enter the
water. Place the nessler cylinder in the right hand side of a nessleriser and match up with the
colours in the standard disc. A white light source should be used. Note the reading.
6. CALCULATION
Dissolved oxygen, ml/l = Reading from the Std disc.

Dissolved oxygen, mg/l = Disc reading x 1.43
GLAB-30 (Revised) - Dissolved Oxygen (Indigo carmine method) Page 3 of 4

Page 117
SECTION: WATER
(WINKLER OR IODOMETRIC METHOD)

Page 118

DISSOLVED OXYGEN (WRINKLER OR IODOMETRIC METHOD)
G-LAB METHOD No.31
(By K.P. Abraham)
1. SCOPE
This test method covers the determination of dissolved oxygen (DO) in boiler feed
waters, recirculated brine etc. The concentration of dissolved oxygen will increase the
corrosivity of these waters.
2. PRINCIPLE
The iodometric test is the most precise and reliable titrimetric procedure for dissolved oxygen
(DO) analysis. It is based on the addition of divalent manganese solution, followed by strong
alkali, to the sample in a glass-stoppered bottle. DO rapidly oxidizes an equivalent amount of
the dispersed divalent managanous hydroxide precipitate to hydroxides of higher valency
states. In the presence of iodide ions in an acidic solution, the oxidized manganese reverts to
the divalent state, with the liberation of iodine equivalent of the original DO content. The
liberated iodine is then titrated with a standard solution of thiosulphate. The titration end point
can be detected visually, with a starch indicator.
3. APPARATUS
3.1. Narrow neck BOD bottles
3.2. Glass stoppers
3.4. Polythene bucket
3.5. Volumetric flask, 500 ml
3.7. Pipettes, 2 ml.
4. REAGENTS
4.1. Manganous sulphate solution (48% W/V)

Dissolve 480 g of manganese sulphate (MnSO4.4H2O) in distilled water and
make up to 1 L.
4.2. Sulphuric acid

Concentrated sulphuric acid (H2SO4 Sp.gr. l.84).
4.3. Alkaline potassium iodide solution

Dissolve 500 g of sodium hydroxide (NaOH) (or700 g KOH) and 135 g of sodium
iodide (NaI) (or 150 g Kl) in distilled water and dilute to 1 L. Add 10 g of sodium azide
(NaN3) dissolved in 40 ml distilled water. Potassium and sodium salts may be used
interchangeably.
GLAB-31 (Revised) - Dissolved Oxygen (Modified Winkler method) Page 2 of 4
Page 119
4.4. Sodium thiosulphate solution, standard (0.01 M)

Dissolve 2.482 g ± 0.001 g of sodium thiosulphate (Na2S203 5H2O) in distilled water
and make upto 1 litre.
4.5. Standard Iodine solution (0.05 M)

To be prepared from BDH Convol, Prod. No. 18038 or equivalent. Transfer
water.

0.5 g of soluble starch is made to a paste with little cold water and then add 25 ml of
boiling water. Boil until a clean solution is obtained.
5. STANDARIDIZATION
Pipette 5.0 ml of the standard iodine solution (0.05 M) to 250 ml Erlenmeyer flask.
Take the prepared sodium thiosulphate solution in a 100 ml burette. Start titration with
constant shaking. When the iodine colour is faint add 2 ml of starch solution and
continue the titration. The end point is noticed by the disappearance of the blue colour.
Note the titre reading.
0.05 M Iodine = 0.10 N Iodine
Normality of Sodium thiosulphate = 0.1 X 5
Titre reading, ml
If the normality obtained differs from 0.01 N, adjust the normality to 0.01 N by adding
calculated amount of Na2S2O3.5H2O or by diluting with distilled water. Standardize again. This
will provide a 0.01 N sodium thiosulphate solution.
6. PROCEDURE
Fill the polythene bucket with sample. Connect a teflon tube to the sample point and drain for
some time to the sample bottle. Immerse the sample BOD bottle in the bucket of water and
continue to flush the bottle with sample and allow it to overflow. Then take out the teflon tube
from the bottle and allow the bottle to remain under water. Keep the glass stopper also under
water. Add 2.0 ml of manganous sulphate solution into the sample using a pipette. Close the
bottle with stopper and mix. Add 2.0 ml alkaline potassium/sodium iodide into the sample
bottle, close and mix. Leave to stand for 5 minutes. Add 2.0 ml sulphuric acid solution into the
sample bottle, close and mix. Remove the sample bottle from water. Take 250 ml of the
solution from the sample bottle in a 500 ml conical flask. Titrate this solution with the standard
thiosulphate solution (0.01 M) using starch as indicator. The end point is noted when the blue
colour disappears to give a colourless solution. Note the titre reading.
7. CALCULATION
Dissolved oxygen, ml/l = Titre reading x 0.224
Dissolved oxygen, mg/l = Titre reading x 0.32
Reference: APHA 4500-0 (C)

ASTM D 888-92 (Reapproved 1996)
Boiler house and power station chemistry by W. Francis.
GLAB-31 (Revised) - Dissolved Oxygen (Modified Winkler method) Page 3 of 4

Page 120
SECTION: WATER
(Using dissolved oxygen test kit)

Page 121

DISSOLVED OXYGEN (USING DISSOLVED OXYGEN TEST KIT)
G-LAB METHOD No.32
(By K.P. Abraham)
1. SCOPE
This test method covers the rapid field determination of dissolved oxygen in brine recycle and
deaerated water. This could cover a range of 0-100 ppb (parts per billion).
2. PRINCIPLE
The sample is filled in the self filling ampoules for the colourimetric analysis of dissolved
oxygen. This ampoule contains a dilute solution of diethylene glycol and a dye. This in
presence of oxygen develops a pink colour which is proportional to the oxygen concentration.
This colour is compared with a comparator to get the oxygen concentration.
3. APPARATUS
3.1. Dissolved oxygen test kit, CHEMetrics, Inv Model 0 - 100.
This kit contains self filling ampoules for the

colourimetric analysis of dissolved oxygen
(CHEMETS R - 7540), Comparator (Type 0-100),
ampoule breaker, funnel, clamp and finger glove.
3.2. Sampling tube
4. REAGENTS
4.1. No separate reagents other than which is supplied with the test kit.
5. PROCEDURE
Connect the sampling tube and the funnel to the sample point. Clamp the funnel at a height
above or parallel to the sample point. Purge the sampling tube and funnel free of air bubbles
with flowing water sample. Sample temperature should be below 200C. Insert the ampoule
breaker into the funnel and allow it to remain there for some time. Then insert a CHEMet into
the ampoule breaker and press to snap the tip. Remove the CHEMet and cover the tip with a
finger, which is covered with the finger glove and mix. Place the sample CHEMet in the centre
tube of the comparator with its tip up. Make comparison by viewing from bottom with top of
comparator directed towards a source of white light. Note the reading corresponding to the
compared colour, as parts per billion dissolved oxgen.
6. CALCULATION
Dissolved oxygen, ppb = Reading from the comparator.
Reference : Manual for dissolved oxygen test kit.
GLAB-32 (Revised) - Dissolved Oxygen (Test Kit method) Page 2 of 2

Page 122
SECTION: WATER
(USING METER)

Date issued : 07thFebruery 1995
Page 123

DISSOLVED OXYGEN (USING METER)
G-LAB METHOD No.33
(By K.P. Abraham)
1. SCOPE
This instrumental method is applicable for the determination of dissolved oxygen in ppb and
ppm levels in water samples, such as steam condensate, deaerated boiler feed water, drinking
water, distillate etc.
2. PRINCIPLE
The water sample is allowed to pass through the flow chamber where the sensor head is
situated. The sensor consists of a cathode of gold and an anode of silver which is being
separated from the outside by a hydrophobic membrane. The oxygen from the water migrates
through the membrane. The anode is held positive with respect to the cathode. Current flowing
through the sensor due to the oxygen reduction at the cathode is converted to a voltage by an
amplifier. The output voltage is proportional to the oxygen concentration, temperature and
membrane permeability. Instrument calibration compensates the membrane permeability and
the temperature is compensated automatically.
3. APPARATUS
3.1. High sensitivity dissolved oxygen meter

Orbisphere 2713 series oxygen analyser, Model 26060
3.2. Sensor
Model 2110
3.3. Calibration cap
3.4. Flow chamber
3.5. Spanner
4. REAGENTS
4.1. Ammonium hydroxide solution

25 % by weight ammonium hydroxide (NH4OH) solution
4.2. Nitric acid
< 70 % by weight concentrated nitric acid
4.3. Recharge kit
Cat No. ..2949.A…
5. CALIBRATION
5.1 System calibration is necessary, before the instrument is used for the first time and
following a membrane change. This calibration should last the life of the membrane.
5.2. Following a membrane change, allow at least 30 minutes for the new membrane to
relax before calibrating the system. Ambient temperatures should be fairly stable
during this time; to confirm, flip the O2/°C switch to °C. Sometimes a sensor must be
serviced in one location and re-introduced to a much colder or warmer area, in which
instances additional time is required for stabilization to occur. The calibration cap is
designed to provide the humidified atmosphere necessary for rapid air calibration.
GLAB-33 (Revised) - Dissolved Oxygen (Using meter) Page 2 of 3
Page 124
5.3. Fill the calibration cap with tap water running at room temperature, then shake out the
water. This leaves the inside wet enough to humidify the air in the chamber.
5.4. Screw the calibration cap loosely into place on the sensor with the sensor coller.
5.5. When the temperature and oxygen readouts are stable, select the 02 mode. Push in and
turn the calibration knob on the front panel, and adjust the display to give the value
10.00. It may take some back-and-forth tweaking of the calibration knob to get all
four digits at this stable reading. Once the value is fixed, release the calibration knob.
This will conclude the calibration.
5.6 Note down the atmospheric oxygen and temperature readings from the display of the
instrument. Now match the readings with the standard oxygen levels at the specified
temperature and normal atmospheric pressure from the following chart.
Temperature °C 20 21 22 23 24 25 26 27 28 29 30
Oxygen ppm 9.05 8.88 8.71 8.54 8.38 8.23 8.08 7.94 7.80 7.66 7.53
If the chart is matching with the temperature and the oxygen concentration readings of
the instrument, it is now ready for measurements.
5.7. Perform a sensor service, when difficulties are experienced with calibration.
6. PROCEDURE
6.1. Replace the protective cap with the sample flow chamber, fix it tightly with the
sensor assembly and admit the sample flow. Adjust the sample flow for a minimum
of 50 ml/min. and maximum of 250 ml/min. The optimum flow is 180 ml/min. When
a stable reading is reached, the display’s rightmost digits may vary in values of three
or four. This slight oscillation is normal. Take the stable reading as the oxygen
concentration.
Note: Sample temperature should not be more than 70 °C. But the desired
temperature is 20 ~ 25 °C for an optimum performance
6.2. After completing the measurements, rinse the sensor with demin water and screw on
the protective cap.
6.3 Keep the instrument switched off but keep it plugged in, to maintain a constant
charge.
6.4. Perform sensor service when:
6.4.1 An unusual long stabilization time for O2 display, either the sensor
exposed to an air saturated media or to changing oxygen
concentration.
6.4.2. Noisy or drifting readouts under a constant oxygen concentrations
Note: Follow the sensor service instruction manual.

7. CALCULATION
Dissolved Oxygen in ppb = Steady reading obtained from the meter.
Result is expressed as <0.010 ppm for values less than 10 ppb and actual values for more
than 10 ppb.
Reference: Manual for oxygen analyser
GLAB-33 (Revised) - Dissolved Oxygen (Using meter) Page 3 of 3

Page 125
SECTION WATER
THE DETERMINATION OF ELIMIN-OX

Date issued : 02nd June 2008
Prepared by : Philip Varkey
Checked by : K.P.Abraham
Page 126

ELIMIN-OX (HACH METHOD)
G-LAB METHOD No.34
(By Philip Varkey)
1. SCOPE
This method covers the determination of ELININ-OX concentration for feed water samples.
Concentration of 0 – 1.5 mg/l can analysis. Reaction glassware should be clean. For best
results, isolate this glassware to be used only for this test.
2. PRINCIPLE
The test is based on iron reduction chemistry and is simple for perform. The ELIMIN-OX in
the sample reduces ferric iron to ferrous iron in direct proportion to the ELIMIN-OX
concentration. This solution then react with Reagent 1 to form a purple color with the ferrous
iron.
3. APPARATUS
3.1. HACH Spectrophotometer -2010

3.2. Pipette 0.50 ml
3.3. Sample testing bottle 50.0 ml
3.4. Sample cell 25.0 ml
4. REAGENTS
4.1. ELIMIN-OX Reagent – 1 100/ pkg (460-S0195.87)

4.2. ELIMIN-OX Reagent -2 100 ml (460-S0196.72)
5. PROCEDURE
5.1 Take two sample testing bottles. One is blank and second is sample.
5.2 Add 25 ml mark to one with Demineralized water and second with sample.
5.3 Add the content of one ELIMIN-OX reagent 1 pillow to each sample testing
bottles. Swirl the cells to dissolve.
5.4 Add exactly 0.50 ml of ELIMIN-OX Reagent 2 to each cell. Mix. Place both
cells in the dark.
5.5 Power on Hach Spectrometer. Enter program # 955 .Adjust the wave length to
562 nm. Press SHIFT + TIMER . A 10 minute reaction period will begin.
GLAB-34 (New) - Eliminox (HACH Method) Page 2 of 3
Page 127
5.6 When the timer beeps Transfer the Blank sample testing bottle content to 25 ml
sample cell. Wipe the cell and keep inside the spectrometer. Close the shield and
press ZERO. 0 mg/l will display. Remove the blank water and transfer sample to
25.0 ml sample cell. Press READ. The result of ELIMIN-OX mg/l will display
6 INTERFERENCES
Substances that reduce ferric iron( such as other oxygen scavengers) can interfere. A
cooler equipped with a copper cooling coil should NOT be used. Trace levels of
copper catalyze the reaction of ELIMIN-OX and oxygen. As a result of copper
contamination, any ELIMIN-OX present in the system may be consumed prior to
analysis.
GLAB-34 (New) - Eliminox (HACH Method) Page 3 of 3

Page 128
SECTION: WATER
THE DETERMINATION OF FERRIC CHLORIDE

Checked by : K.P. Abraham
Page 129

FERRIC CHLORIDE
G-LAB METHOD No.35
(By Philip Varkey)
1. SCOPE
This method is used for the determination of ferric chloride concentration in the bulk
chemical supply.
2. PRINCIPLE
Iodometric titration with standerdised sodium thiosulphate solution using starch as
indicator. At the end point colour change from blue to colourless solution.
3. APPARATUS
3.1 Conical flask
3.2 Burette
4. REAGENT
4.1 Sodium thiosulphate (0.1 N)
4.2 Potassium Iodide
4.3 Starch ( 1 % solution)
5. REAGENT PREPERATION
5.1 Sodium thisulphite ( 0.10 N)
Dissolve 24.82 g of Sodium thiosulphate (Na2S2O3.5H20 ) in boiled and then
cooled distilled water. Dilute to 1 L and add 0.10 g of Sodium carbonate as a
preservative. This solution deteriorates on standing and should be standardized
every week, using standard Iodine solution.
5.2 Starch (1 % Solution)

0.50 g of soluble starch is made to a thin paste with a little cold water and
then, add 25 ml of boiling water. Boil until a clean solution is obtained. This
should be freshly prepared as required.
GLAB-35 (New) - Ferric chloride Page 2 of 3

Page 130
6. PROCEDURE
Weigh, Ferric chloride (~ 0.50 grams ) in a weighing boat and transfer
quantitatively to a conical flask. Dilute to 100 ml with DM water and add
5 grams of potassium iodide. Titrate against standard Sodium thisoulphate
( 0.1 N). When the brown colour reduces, add 2.0 ml of starch solution
and continue titratation till the blue colour disappears. Note the titre reading at
colour change as V.
7. CALCULATION
Ferric Chloride, % w/w = 0.10 x V x 162.18 x 100

W
Where,
0.10 = Normality of Sodium thiosulphate
V = Titre reading (Volume of Sodium thisulphate consumed)
W = Weight of Ferric chloride in milligram.
162.18 = Equivalent weight of Ferric chloride
GLAB-35 (New) - Ferric chloride Page 3 of 3

Page 131
SECTION : WATER
THE DETERMINATION OF HARDNESS
(TOTAL, CALCIUM AND MAGNESIUM HARDNESS)

Date issued : 20thMarch1995
Page 132

TOTAL, CALCIUM AND MAGNESIUM HARDNESS
G-LAB METHOD No.36
(By K.P. Abraham)
1. SCOPE
This method covers the determination of hardness in water by titration. This method is
applicable to raw water, treated water and boiler water those are clear in appearance and tree of
complexing treatment chemicals. The lower detection limit of this method is approximately 0.5
mg/l as CaCO3 the upper limit can be extended to all concentrations by sample dilution. It is
possible to differentiate between hardness due to calcium ions and that due to magnesium ions
by this method.
2. PRINCIPLE
Ethylene diaminetetraacetic acid and its sodium salts (EDTA) form a chelated soluble complex
when added to a solution of certain metal cations. If a small amount of a dye such as Erichrome
Black T is added to an aqueous solution containing calcium and magnesium ion at a pH of 10.0
+ 0.1, the solution becomes wine red. If EDTA is added as a titrant the calcium and magnesium
will be complexed, and when all of the magnesium and calcium has been complexed, the
solution turns from wine red to blue, marking the end point of titration. Magnesium ions must
be present to yield a satisfactory end point.
When EDTA is added to water containing both calcium and magnesium, it combines first with
the calcium. Calcium can be determined directly, with EDTA when the pH is made sufficiently
high then the magnesium is largely precipitated as the hydroxide and an indicator is used that
combines with calcium only. For the detection of end point, two methods are available. In
method—A, murexide (ammonium purpurate) gives a colour change when all the calcium has
been complexed by EDTA at a pH of 12 to 13. This indicator changes from pink to purple at
the end point. In method—B, Nana indicator (calcon carboxyl acid) gives a colour change
when all calcium has been complexed by EDTA at pH of 12 to 13. This indicator changes from
reddish purple to blue at the end point.
3. APPARATUS
3.2, Conical flask, 250 ml
3.3. Measuring cylinders, 100 ml and 50 ml
3.4. Volumetric flasks, 1 litre and 500 ml
4. REAGENTS
4.1. Disodium ethylenediamine tetra acetate (Na2H2EDTA) solution Standard (0.01M).
Weigh 3.723 g analytical reagent grade disodium ethylenediarninetetra acetate

dihydrate (EDTA) and dissolve in distilled water and dilute to 1000 ml. Standardize
against standard calcium or zinc chloride solution.
GLAB-36 (Revised) - Hardness (Total, Calcium & Magnesium) Page 2 of 4

Page 133
4.2. Standard calcium solution (1 ml = 1 mg CaCO3)

Weigh 1.000 g anhydrous CaCO3 powder (primary standard or special reagent low in
heavy metals, alkalies and magnesium) into a 500 ml erlenmeyer flask. Place a funnel
in the flask neck and add, a little at a time, 1+1 HC1 until all CaCO3 has dissolved.
Add 200 ml distilled water and boil for a few minutes to expel CO2. Cool, add a few
drops of methyl red indicator, and adjust to the intermediate orange colour by adding
3N NH4OH or 1+1 HC1, as required. Transfer quantitatively and dilute to 1 litre with
distilled water.
4.3. Ammonia buffer (pH 10)
Dissolve 67.5 g ammonium chloride (NH4C1) in 570 ml of ammonia solution
(Sp.gr.0.91). Make up to 1 L using distilled water.
4.4. Calcium hardness buffer (pH 12—13)
Take 300 ml distilled water and add 225 g potassium hydroxide (KOH) and dissolve.
Make up to 500 ml.
4.5. Total hardness indicator
Grind 0.5 g of solochrome black (Erichrome black, mordant black II) with 100 gm of
powdered sodium chloride. Use dark coloured bottle for storage.
4.6. Erichrome black—T indicator solution (EBT indicator solution) Dissolve 0.5 g of
solochrome black (Erichrome black,mordant black II) in 100 ml of methyl alcohol,
4.7. Nana indicator
Calcon carboxyl acid (C21H14O2N2S)
4.6. Murexide indicator
Prepare by mixing 0.5 g murexide (ammonium purpurate) with 100 g solid sodium
chloride and grinding the mixture to 40 to 50 mesh.
5. STANDARDIZATION
Pipette 10.0 ml of the standard calcium solution (1 ml = 1 mg CaCO3) to 250 ml conical flask.
Add some 90 ml distilled water to it. Add 2 ml of ammonia buffer. Add a pinch of total
hardness indicator or 2 drops of EBT indicator solution and mix. Take the EDTA solution in
the burette. Titrate the calcium solution with EDTA from the burette, until the colour changes
from wine red to blue. Note the titre reading.
Normality of EDTA = _____ 2_______

Titre reading
Adjust the normality of the EDTA solution by adding calculated amount of EDTA or diluting
with distilled water. The final titre reading should be 10.0 ml for the 10.0 ml calcium standard
solution.
6. PROCEDURE
6.1. Total hardness
Take 100 ml sample. Add 2 ml of ammonia buffer. Add 2 drops of EDT indicator
solution or a pinch of total hardness indicator powder and mix. Titrate with the
standard EDTA solution until the colour changes from wine red to blue.
Note the titre reading as TH’.
6.2. Calcium hardness
Method - A
Take 100 ml sample. Add 4 ml of caustic buffer. Add a pinch of murexide
indicator and mix. Titrate with the standard EDTA solution until the
colour changes from pink to purple. Note the titre reading as ‘TC’.
Method - B
Take 100 ml sample. Add 4 ml of caustic buffer mix, wait and let stand for
5 minutes. Add a pinch of Nana indicator and mix. Titrate with the
standard EDTA solution until the colour changes from reddish purple to
blue. Note the titre reading as ‘TC’
Page 134
7. CALCULATION
Total hardness, mg/l as CaCO3 = ____TH x 1000___

Vol.of sample ml
Calcium hardness, mg/l as CaCO3 = ____TC x 1000____

Vol. of sample ml
Magnesium hardness mg/l as CaCO3 = Total hardness - Calcium hardness.
Note:
TH = Titre reading for total hardness, TC = Titre reading for calcium hardness.

APHA 2340 (C) & 3500 Ca (D)

Page 135
SECTION : WATER
THE DETERMINATION OF HYDRAZINE

Page 136

HYDRAZINE IN BOILER AND BOILER FEED WATER
G-LAB METHOD No.37
(By K.P. Abraham)
1. SCOPE
This method covers the colourmetric determination of hydrazine in water. Hydrazine is a man-
made chemical and is not found in natural waters. The determination of hydrazine is usually
made on boiler feed waters, process waters and other waters that have been treated with
hydrazine, for the purpose of maintaining residuals to prevent corrosion by dissolved oxygen.
2. PRINCIPLE
Para - dimethylaminobenzaldehyde produces a specific, yellow reaction product with
hydrazine. The intensity of the yellow colour is proportional to the amount of hydrazine in the
water and follows Beer’s law.
3. APPARATUS
JASCO, Model V 530
3.2. Sample cells 10 mm cells
3.3. Pipettes 1, 2, 4, 5 and 10 ml
3.5. Volumetric flasks, 50 ml, 7 Nos.
3.6. Beakers, 100 ml, 10 Nos.
4. REAGENTS
4.1. Para-dimethylaminobenzaldehyde solution (P-DMAB reagent). Dissolve 40 g of Para-

dimethylaminobenzaldehyde in 1L of 2N sulphuric acid.
4.2. Hydrazine solution, standard (lml = 1mg N2H4) (Stock) Dissolve 4.063 g of hydrazine
sulphate (N2H4.H2S04) in distilled water and make upto 1 litre.
4.3. Hydrazine solution, standard (lml = 0.005 mg N2H4)
Pipette 5.0ml of the standard hydrazine solution (lml = 1mg N2H4) to a 1 litre
volumetric flask and make up to the mark with distilled water.
5. CALIBRATION
Pipet 1.0,2.0, 4.0, 6.0, 8.0 and 10.0 ml of the standard hydrazine solution (l ml = 5 µg N2H4) to
50 ml volumetric flask and make upto the marks. This will give standard solutions of 0.10,0.20,
0.40, 0.60, 0.80 and 1.00 mg/l of hydrazine. Transfer these to 100 ml beakers. Take 50 ml
distilled water as blank. Add 5 ml each of the para-dimethylaminobenzaldehyde solution to
each and mix. Wait for 30 minutes and measure the absorbance at 456 nm wavelength using 10
mm cells against distilled water reagent blank. Plot a graph with absorbance against
concentration (mg/l N2H4). Calculate the factor from the graph, which is equal to mg/l N2H4
required for unit absorbance.
Note: In G - Station laboratory, this calibration graph and analytical parameters are stored in
File No.2 of the JASCO spectrophotometer.
GLAB-37 (Revised) - Hydrazine (Colorimetric method) Page 2 of 3

Page 137
6. PROCEDURE
Take 50 ml sample and 50 ml distilled water blank in beakers/cups. Add 5 ml

p-dimethy aminobenzadehyde reagent to each, mix and wait for 10 minutes. Measure the
absorbance at
456 nm using 10 mm cells, against the distilled water plus reagent blank. Note
the absorbance.
For G-Station laboratory, load the file in JASCO spectrophotometer and the concentration can
be measured directly.
7. CALCULATION
Hydrazine, mg/l N2H4 = Sample absorbance x Factor from the calibration graph
GLAB-37 (Revised) - Hydrazine (Colorimetric method) Page 3 of 3

Page 138
SECTION WATER
THE DETERMINATION OF HYDRAZINE
(IODIMETRIC Method )

Page 139

HYDRAZINE
[IODIMETRIC Method ]
G-LAB METHOD No.38
(By Philip Varkey)
1. SCOPE
This method covers the determination of Hydrazine for high concentration. This method is
applicable during the wet preservation & boiler hydro test time of boilers.
2. PRINCIPLE
Hydrazine reacts with iodine in the presence of sodium bicarbonate (NaHCO3) according to
the following formula.
N2H4+2I2 +4NaHCO3 ► N2 + 4NaI +4H20 +4H20
Hydrazine is titrated with iodine solution using starch as the colour indicator.
3. APPARATUS
3.1. Pipette
3.2. Burette
3.3. Conical Flask
4. REAGENTS
4.1. Iodine standard solution (0.10 N)

4.2. Sodium Bicarbonate.
4.3. Starch ( 1 % ) solution
5. Reagent preperation
5.1. Iodine standard solution (0.10 N)
To be prepared from the BDH CVS (Prod.No.18038) or equivalent. Transfer
quantitatively to a 1 Liter volumetric flask and make up to the mark with
distilled water.
0.50 g of soluble starch is made to a thin paste with a little cold water and then
add 25 ml of boiling water. Boil until a clean solution is obtained. This should
be freshly prepared as required.
GLAB-38 (New) - Hydrazine (Iodimetric method) Page 2 of 3

Page 140
6. PROCEDURE
6.1. Take adequate volume of sample into a conical flask.

6.2. Add about 0.50 g of sodium bicarbonate and 3.0 ml of starch solution into
the sample with mixing.
6.3. Titrate the sample against 0.10 N iodine standard solution, until the blue
starch iodine colour appears.
7. CALCULATION
Calculate the Hydrazine concentration as follows

Hydrazine, ppm = A x 1000x 0.80
V
Where,
A = ml of iodine standard solution required for sample titration
V = ml of sample taken
GLAB-38 (New) - Hydrazine (Iodimetric method) Page 3 of 3

Page 141
SECTION : WATER
THE DETERMINATION OF HYDROCHLORIC ACID CONCENTRATION

Page 142

HYDROCHLORIC ACID CONCENTRATION IN ACID CLEANING SOLUTION
G-LAB METHOD No.39
(By K.P. Abraham)
1. SCOPE
This test method is used for the determination of the hydrochloric acid concentration (HC1 %)
in acid cleaning solution. It is essential to determine the acid strength to control the acid clean
process and to asses the degree of cleanliness.
2. PRINCIPLE
A measured quantity of the sample is titrated with a standard sodium hydroxide solution
(NaOH, 1M), using methyl orange as an indicator.
3. APPARATUS
3.1. Erlenmeyer flask, 250 ml
3.2. Measuring cylinder, 50 ml & 100 ml
3.4. METTLER DL25 Titrator
3.5. Indicator bottle
4. REAGENTS
4.1. Sulphuric acid, standard (0.5M)

To be prepared from a BDH Convol Prod. 18045 or equivalent. Transfer
water.
4.2. Sodium hydroxide, standard (1M)

Dissolve 40.0 g of sodium hydroxide (NaOH pellets) in 400 ml distilled water cool
and dilute to 1 L Standardize using the standard 0.5M sulphuric acid solution.
4.3. Methyl orange indicator.

Dissolve 0.05 g of methyl orange in water and dilute to 100 ml.
Dissolve 1 g of phenolphthalein in 60 ml alcohol and then add 40 ml distilled water

and mix.
GLAB-39 (Revised) - Hydrochloric acid concentration Page 2 of 3

Page 143
5. STANDARDIZATION
Pipette 10.0 ml of the prepared sodium hydroxide solution (1M) to a 250 ml Erlenmeyer flask.
Add 5 drops of phenolphthalein indicator. Take the standard sulphuric acid (0.5M) in a burette
and titrate with the sodium hydroxide solution. The end point is noticed by the disappearance
of the pink colour. Note the titre reading.
0.5M H2S04 = 1N H2S04
Normality of Sodium hydroxide = 1 x titre reading

10
If the normality obtained is found differing from 1N, adjust the normality to 1N by adding
calculated amount of NaOH or by diluting with distilled water. Standardize again.
This will provide a 1M NaOH solution.
6. PROCEDURE
Take 50 ml of the sample in a 250 ml Erlenmeyer flask. Add 5 drops of methyl orange
indicator. Titrate the sample solution with the stapdard sodium hydroxide solution (1M), until
the colour changes from red to orange. Note the titre reading.
In G - Station laboratory, autotitrator can be used for the titration. By calling method No.4 in
METTLER DL25 titrator, all the parameters required for the autotitration method will be set.
No indicator is necessary when using autotitrator. Follow the analytical procedures for the
autotitrator. On completion of the titration the acid concentration will be displayed as well as
printed.
7. CALCULATION
Hydrochloric acid % = Titre reading x 0.073
Where,
0.073 = __Mole Wt. of HC1 x 1 = 36.5 x _1_
Sample volume ml x10 50 10
(1/10 is for converting g/l to %)
GLAB-39 (Revised) - Hydrochloric acid concentration Page 3 of 3

Page 144
SECTION: WATER
THE DETERMINATION OF IODINE NUMBER OF ACTIVATED CARBON

Page 145

IODINE NUMBER OF ACTIVATED CARBON
G-LAB METHOD No.40
(By Philip Varkey)
1. SCOPE
This test method covers determination of relative activated level of unused or
reactivated carbons by absorption of iodine from aqueous solution. The amount of
iodine absorbed (in milligrams) of carbon using test conditions listed herein is called
the iodine number.
2. PRINCIPLE
A standard iodine solution is treated with activated carbon under specific conditions.
The carbon treated solutions are filtered to separate the carbon from the treated iodine
solution (filtrate) iodine remaining in the filtrate is measured by titration. The amount
of iodine adsorbed (in milligrams) per gram of carbon at a residual iodine
concentration of 0.05 N is reported as the iodine number.
3. APPARATUS
3.1 Pipette
3.2 Burette
3.3 Beaker
4. REAGENT
4.1 Sodium Thisoulphate (0.05N)
4.2 Iodine (0.05N)
4.3 Starch solution (1%)
5. REAGENT PREPARATION
5.1. Sodium Thiosulphate (0.05 N)
Dissolved 12.412 g of sodium Thiosulphate (Na2S2O3.5H20 ) in boiler and cooled
distilled water. Dilute to 1 L and add 0.10 g of sodium carbonate as a preservative.
This solution deteriorates on standing and should be standardized every week, using
standard Iodine solution.
GLAB-40 (New) - Iodine number of Activated Carbon Page 2 of 3

Page 146
5.2 Iodine solution, standard (0.05N)

quantitatively to a 1 Liter volumetric flask and make up to the mark with distilled
water
6. PROCEDURE:
6.1. Grind the sample and make fine power still 95 % by weight will pass through
100 mesh screen
6.2. Take ~ 0.05 gram (Exact weight ) in a 100 ml of beaker. Note the weight as
(W) grams
6.3. Add 40 ml of 0.05 N Iodine solution to the sample.
6.4. Keep the sample on the stirrer and stir for one minute
6.5. Decant the iodine solution to a clean beaker
6.6. Take 25 ml of the decant solution from the beaker in conical flask.
6.7. Titrate against 0.05 N Sodium thiosulphate using starch as indicator. Note the
titre volume when the solution becomes colourless as (S) ml.
6.8. Take 25 ml of 0.05 N original iodine solutions and titrate against 0.05 N
Sodium thisoulphate with starch as indicator. Note the titre volume when the
solution becomes colourless as (B) ml. This will be the blank titre volume.
7. CALCULATION
Iodine Absorption Number, g /Kg = [(B-S)/B] x (V/W) x N x 126.91
Where,
B = Sodium thiosulphate required for the blank, ml
S = Sodium thiosulphate required for the sample, ml
V = Sample volume, ml
W = Weight of the sample, g
N = Normality of iodine solution (0.05N)
126.91 = Equivalent mass of Iodine
GLAB-40 (New) - Iodine number of Activated Carbon Page 3 of 3

Page 147
SECTION : WATER
ION CHROMATOGRAPHY, ANIONS

(F-,Cl- ,NO2- ,Br - ,NO3- ,P04-- ,SO4--)

Date issued : 20thJanuary 1996
Page 148

ION CHROMATOGRAPHY, ANIONS (F-,Cl-,NO2-,Br-,NO3-,PO4 --,SO4--)
G-LAB METHOD No.41
(By K.P. Abraham)
1. SCOPE
This test method covers the sequential determination of fluoride, chloride, nitrite, bromide,
nitrate, ortho-phosphate, and sulphate in water by chemically suppressed ion chromatography.
This test method is applicable for drinking water, waste water, cooling water, boiler water,
steam, sea water etc. Ion chromatography provides for both qualitative and quantitative
determination of seven common anions in the mg/L range from a single anlytical operation
requiring a few milliliters of sample and taking approximately 10 minutes for completion.
Anion combinations such as Cl-/Br- and NO2- /NO3-, which may be difficult to distinguish by
other analytical methods, are readily separated by ion chromatography.
2. PRINCIPLE
An aliquot of sample is injected into an ion chromatograph. The sample is pumped through two
columns and a suppressor device and into a conductivity detector. The analytical column and
the guard column are packed with low—capacity anion exchanger. Ions are separated based on
their affinity for the exchange sites of the resin. The suppressor device contains a membrane
based cation exchanger which is continuously regenerated by a flow of dilute sulphuric acid.
The suppressor device reduces the background conductivity of the eluent to a low or negligible
level by replacing the cations with the hydrogen ion, thereby converting anions in the sample to
their corresponding acids. The separated anions in their acid form are measured using an
electrical conductivity cell. Anions are identified based on their retention times compared to
known standards. Quantitation is accomplished by measuring the peak height or area and
comparing it to a calibration curve generated from known standards.
Interference
Since chloride and nitrite elute very close together, they are potential interferents for
each other. It is advisable not to have one of these anions present in a ten—fold excess
over the other; that is, Cl-/NO2- ratios higher than 1:10 or 10:1 if both ions are to be
quantitated.
Water from the sample injection will cause a negative peak or dip in the chromatogram
when it elutes, because its conductivity is less than that of the suppressed eluent. This
dip usually occurs before Cl-. Any peak of interest (eg. F-) eluting near the water dip
must be sufficiently resolved from the dip to be accurately quantitated. Addition of 1%
v/v concentrated eluent with the sample is recommended in such cases to overcome the
water dip.
3. APPARATUS
3.1. Ion Chromatograph, DIONEX DX—100

3.2. Guard column, IonPac AG4A—SC 4 mm
3.3. Analytical column, IonPac AS4A—SC 4mm
3.4. Sample loop, 25 µl
3.5. Suppressor, Anion micro membrane suppressor,
Model AMMS—II
3.6. Integrator, Chromjet SP 4400 by Spetra—Physics
3.7 Mixed bed De - ionized water assembly, Milli-Q Gradient System
GLAB-41 (Revised) - Ion Chromatography (An ions) Page 2 of 6

Page 149
4. REAGENTS
4.1. Double De-ionized water
Fresly prepared De-ionnized water from WATER I with resistance
>17 megohm-cm.
4.2. Eluent ( 2.2 mM Na2CO3 / 2.8 mM NaHCO3)
Dilute AS3 / AS4 eluent concentrate by 100 times.
Alternatively this can be prepared from solid analar grade chemicals.
Dissolve 0.4704 g of Sodium bicarbonate (NaHCO3) and 0.4664 g of
Sodium carbonate (Na2CO3) in De-ionized water and make upto 2 L
with De-ionized water.
4.3. Suppressor reagent (0.05N H2S04)
Dilute 100 ml of 1 N sulphuric acid to 2 L with DM water.
4.4 Stock solutions of standards.
Use ready made stanards of F-, Cl-, NO2-, Br-, NO3-, P04- -, SO4- - of
1 mg/ml concentration. Dilute these standards to the required
concentrations to prepare the standard mixtures. Use De-ionised water
for the dilutions.
4.5 Standard mixtures
Prepare standard mixtures as specified in Table—I
Table - I
-------------------------------------------------------------------------------------------------------------------------
Anion/STD No. 1 2 3 4 5 6 7 8 9 10 11
--------------------------------------------------------------------------------------------------------------------------
Fluoride mg/L 2 5 10 0.1 0.5 1.0 0.2 0.8 1.5 — —
Chloride mg/L 2 5 10 50 150 400 75 150 300 0.1 0.5
Nitrite mg/L 2 5 10 — — — 2 5 10 — —
Bromide mg/L 2 5 10 0.4 0.8 1.5 0.5 1.0 2.0 0.05 0.1
Nitrate mg/L 2 5 10 0.1 0.5 1.0 50 100 200 — —
Phosphate mg/L 2 5 10 0.5 1.0 2.0 5 10 20 1 2
Sulphate mg/L 2 5 10 10 20 40 10 20 40 0.1 0.2

-----------------------------------------------------------------------------------------------------------------------------------
Anion/STD No. 12 13 14 15 16 17 17A 18 19 20 21
-----------------------------------------------------------------------------------------------------------------------------------
Fluoride mg/L — — — — — — — — — — —
Chloride mg/L 1.0 0.01 0.02 0.04 0.05 0.5 10 20 ------- ~ 500 --------
Nitrite mg/L — — — — 40 100 200 200 — — —

Bromide mg/L 0.2 0.01 0.02 0.04 0.05 0.1 0.15 0.2 0.5 1.5 3.0
Nitrate mg/L — — — — 10 50 75 100 — — —
Phosphate mg/L 4 0.01 0.02 0.04 0.1 0.2 0.40 0.5 — — —
Sulphate mg/L 0.4 0.01 0.02 0.04 1 2 3 5 ---------- ~ 65-----------

-----------------------------------------------------------------------------------------------------------------------------------

Page 150
5. CALIBRATION
Prepare a file for the analysis by entering all the standard datas and the analytical and integrator
conditions through the dialog. Use three calibration standards as specified in Table —II for each
file. Complete the calibration by injecting the standards in the sequence in which the standards
are entered in the file. Follow the analytical procedure given below for the standards and
samples.
Routine analysis procedure
5.1. Check the Eluent and Suppressor solutions. If the level is less than 25 %, then fill the
reservoir with fresh solutions.
(i) Eluent — 2.2 mM Na2CO 3 / 2.8 mM NaHCO3
(ii) Suppressor — 0.05N H2S04
(iii) Rinse — Double demin. water
5.2. Open the Helium gas supply valve. (Keep pressure 7 bar)
5.3. Switch ON the Ion Chromatograph and set the control button on the system panel to Local.
5.4. Switch ON the eluent nitrogen pressure switch. (Keep pressure 7 ± 2 psi)
5.5. Make the dual channel control switch to the following positions: Elu A/B, Elu A, and Syringe.
5.6. Set the pressure Hi limit to 2000 psi.
5.7. Make the Low limit to OFF.
5.8. Open the pump priming valve by two turns and start the pump.
5.9. When there is no more air bubbles is seen coming through the drain, close the priming valve.
5.10. Set the eluent flow rate to 2.67 ml/min (pump dial setting 120) and suppressor flow rate
to 4.0 ml/min. (pressure 5.0 psi)
5.11. Select the conductivity range according to the method No. to be used.
5.12. Wait till the conductivity reading stabilizes. (16 ~ 17 µS/cm)
5.13. Switch ON the Auto offset and the Low limit ON.
5.14. Observe the level reading in the integrator.
5.15. Select the method by selecting the file number. Type FI: (file number) and press enter.
5.16. Set the anlytical conditions according to the method No. selected, from Table — II.
5.17. Make the sample button to Load position.
5.18. Take the sample in the syringe provided and push it through the injection valve.
5.19. Purge like this two times.
5.20. Press the sample button and immediately press the INJ A button in the integrator.
5.21. Sample will be analysed and the report will be printed.
5.22. Wait for 15 minutes and then switch OFF the pump.
5.23. Switch OFF the eluent nitrogen pressure switch.
5.24. Shut OFF the Helium gas supply.
5.25. Release the line pressure by opening the suppressor reservoir relief valve.
5.26. Switch OFF the Ion Chromatograph.
CAUTI ON
NEVER SWITCH OFF THE ON/OFF SWITCH IN THE INTEGRATOR.

Page 151
NOTE:
(i) For rinsing the system, make pump pressure low limit to OFF and open the
priming valve and change the selector from Elu A/B to Rinse in the dual channel
control panel. If the pump is not running, start the pump. When there is no air
bubble is seen coming through the drain, close the priming valve and wait for 15
minutes. This type of rinsing is required when the column is suspected to be
contaminated with high levels of ions.
(ii) Long time storage of the column and TAC should be done with 0.1M NaOH.
6. PROCEDURE
6.1. Filtre the sample with 0.45 µm filter paper if it contains any suspended particles.
6.2. For concentrated samples, dilute the sample with DM water to bring it to the
calibration standards range.
6.3. Follow the routine analytical procedure for the sample measurement.
7. CALCULATION
The concentration of each an ion will be calculated and printed by the integrator after
completing the plotting of the chromatogram.
Reference:
ASTM D4327
Manual for Dionex DX—100

Page 152
ION CHROMATOGRAPH ANALYTICAL CONDITIONS
FOR DIFFERENT DEWA METHODS
Table II —
1. DEWA Method # 1 2 3 4 5 6 7
2. File No,(FI) 1 2 3 4 5 6 7
3. Analysis for STD 2/5/10 DRNK.WATER SEWG. WATER BLR. WATER SATRD STEAM CLNG WATER SWTR Br-
4. Conductivity range (µS/cm) 100 1000 1000 3 1 300 10
5. Peak Threshold (PT) 200 50 50 2000 20000 100 1000
6. Attenuation (AT) 512 512 512 512 512 512 1024
7. Standards used for calibration 1,2,3 4,5,6 1,8,9 10,11,12 13,14,15 16,17,17A,18 19,20,21
8. Lower and higher limits, EL/EH - F - 1/15 0.05/1.5 0.1/2.0 -/- -/- -/- -/-
Lower and Higher limits, EL/ER Cl _ 1 / 15 25/ 500 50/350 0.05 /1.5 0.01 / 0.05 0.2/250 -/-
Lower and Higher limits, EL/EH NO2 -/- 1/15 -/- -/- 500/3500 10/175
Lower and Higher limits, EL/ER Br- 1 / 15 0.2 / 2.0 0.2/2.5 0.02 / 0.30 01/ 0.05 0.3/2,5 -/-
Lower and higher limits, EL/EN NO3 1 / 15 0.05/ 1.5 25/250 - /- -/- 50/ 1200 -/ -
Lower and Higher limits, EL/EN P04- 1 / 15 0.2/2.5 2.0/25 0.5 / 5.0 0.01/0.05 0.5 / 1.0 -/-
Lower and Higher limits, EL/ENSO4 1 / 15 5.0/50 5.0/50 0.05/0.5 0.01/ 0.05 5.0/60 -/ -
9. Addition of concentrated eluentl % v/v Yes(**) Yes(**) Yes(**) No No Yes(*) No
10. Blank chromatogram (Bin) subtraction No No No Yes Yes No No
11. Sample dilution (Times) (XF) 1.0(***) 1,0(****) 1.0(****) 1.0 1.0 10 50
* Eluent addition — 100 ml STD/Sample + 1 ml Con. Eluent
** Eluent addition — 99 ml STD/Sample + I ml Con.Eluent
*** To be changed to 0.99 if con.eluent is not added in the sample.
**** To be changed to 1.01 if con.eluent is added in the sample.

Page 153
SECTION : WATER
ION SELECTIVE ELECTRODE DETERMINATIONS

(pH, F-,NO3-,K+,Ca ++,Na+)

Date issued : 21stDecember 1995
Page 154

ION SELECTIVE ELECTRODE DETERMINATIONS (pH, F-, NO3 --, K+, Ca++, Na+)
G-LAB METHOD No.42
(By K.P. Abraham)
A. OPERATION OF WTW pMX 2000 pH / ION METER
1. SCOPE
This instrument is useful in the measurements of pH and other selective ions using specific ion
electrodes. The accuracy of the analyses are undisturbed by the colour or the turbidity of the
samples.
2. PRINCIPLE
The operation of the WTW pMX 2000 meter is always a dialogue between the user and the
instrument. This means that the user is always guided step by step by the instrument.
2.1. pH measurements are as usual using a pH glass electrode having automatic temperature
control and specific ions are tested using separate electrodes, reference electrode and
temperature compensators. The technics of measurement of specific ions vary each
other. It will be explained in individual procedures.
To enable to execute different technics of measurements there are twenty two built in
programs stored in the instrument memory.
They are:-
2.2. Program — 0
This program is used to limit the values, which are being measured. When the limited
values are exceeded in the measurements it will give an acoustic and optical signal.
2.3. Programs for pH measurements: —

2.3.1. Program — 1
MEMO CAL - pH calibration.
This program is used to calibrate the pH meter using buffers other than the
specified DIN/NBS or stored technical buffers. Since the instrument memory
is not aware of the temperature behaviour of the new set of buffers the values
must be put in dependence on the temperature.
2.3.2. Program — 2
ISO CAL - pH calibrations.
This program is used to get exact temperature compensation. Select any one
of the two sets of the instrument specified buffers (DIN—NBS/technical) and
calibrate the instrument with two different temperatures.
2.3.3. Program — 3
Single point calibration.
If the slope of your pH electrode is known, the calibration can be done with
one buffer only.
2.3.4. Program — 4
Indication and input of electrode data.
This program enables you to view/change/over write latest program stored in
the memory of the instrument.
GLAB-42 (Revised) - Ion Selective Electrode determinations Page 2 of 23

Page 155
2.3.5. Program — 5
Input of the sample temperature.
If the temperature compensation is to be given externally measure the sample
temperature externally and enter the values.
2.4. Programs for ion selective measurements: —
2.4.1. Program — 6
Two point calibration for direct potentiometry in the linear range
The instrument calculates the calibration line by the results of 2
measurements in calibration standard solutions and indicates the electrode
slope at the end of the calibrations. The measurement is effected by program
— 12.
2.4.2. Program — 7
Single point calibration for direct potentiometry in the linear range.
If the slope of the electrode is known calibration can be done using one buffer
solution only.
2.4.3. Program — 8
Input of known electrode slope without any calibrations.
2.4.4. Program — 9
Determination of electrode slope.
When the electrode is immersed in two different standards having a
concentration difference of one decade (eg..01 & .001) the
instrument reads a stable reading and the slope is accepted
automatically.
2.4.5. Program — 10
Calibration for direct potentiometry in the total range of the characteristics.
This program is used to measure even a small concentration starting from the
non linear to linear range. Six standards are required for the calibrations. Five
should be of non linear range where as standard 5 should be at the junction
line between the non linear and the linear range. Standard 5 and 6 then limit
the linear part of the measuring range. Measurement is effected by program
— 20.
2.4.6. Program — 11
Determination of blank value.
Blank value for TISAB is determined by this program.
2.4.7. Program — 12
Direct potentiometry in the linear range.
Used for the measurements of programs 6 or 7.
2.4.8. Program — 13
Known addition.
This program can be used for measurements of samples of higher salt
concentrations.
2.4.9. Program —14

Known addition with blank value correction
Program for small concentrations. Blank is decided by program 11 and is
being deducted.

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2.4.10. Program — 15
Known subtraction
The instrument calculates the concentration to be subtracted when the added

standard is part of the substance to be tested.
2.4.11. Program — 16
Sample addition.
This method is the inversion of the known addition method i.e. a standard is
measured first and the sample is added and measured again.
2.4.12. Program — 17
This is a method for the determination of thorium, uranium or zirconium.

The sampleis pipetted to a given standard volume whereby the indicator
decreases the concentration of the standard.
2.4.13. Program — 18 & 19

Sample addition and subtraction program.
Programs 15 & 16 are modified to suit the addition of solid samples.
2.4.14. Program — 20
Direct potentiometry in the total range of characteristics.
This is the measurement program of program 10.
2.4.15. Program — 21
Double standard addition.
For this method neither a calibration nor a slop is required for the
measurement. It is suitable for any difficult sample material. The results are
fixed addition relations of 1% for the first known addition (sample — 2) and
2% for the second known addition (sample — 3).
Reference: pH/ION meter manual.

Page 157
B. DETERMINATION OF pH
1. SCOPE
Measurement of pH is one of the most important and frequently used tests in water chemistry. At
a given temperature intensity of the acidic or basic character of a solution is indicated by pH or
hydrogen ion activity.
2. PRINCIPLE
2.1. Definition: The pH of a solution is the negative common log of the hydrogen ion activity
a H+. pH = -log [a H+]
2.2. The glass electrode is the most widely used hydrogen ion responsive electrode. The glass
electrode available is a combination electrode which contain an indicator electrode (a
thin glass bulb) and a reference electrode (silver—silver chloride) combined in a single
unit. The thin glass bulb and the narrow tube attached to it are filled with 0.1M HC]. and
carry a silver—silver chloride electrode in it. The external tube is fused to the lower end
of the narrow tube. The external tube is filled with saturated KC1 which is also saturated
with silver chloride and carries a silver—silver chloride electrode in it. The assembly is
sealed with an insulating cap.
2.3. pH of a solution is derived from the e.m.f. of the cell

GLASS ELECTRODE |SOLUTION| |REFERENCE ELECTRODE. The measurement
of the concentration of hydrogen ions using a pH meter is clearly an example of direct
potentiometry. When the electrode is immersed in a solution a potential is developed at
the glass surface, which is proportional to the hydrogen ion activity of the given
solution. The operation of a glass electrode is related to the situation existing at the inner
and outer surfaces of the glass membrane. Glass electrodes require soaking in water for
some hours before use and a hydrated layer is formed on the glass surface, where an ion
exchange process can take place. If the glass contains sodium it will be replaced by
hydrogen ion in an equilibrium reaction. Since the contents of the indicator electrode are
sealed and the concentration of the internal HC1 is constant the potential of the silver—
silver chloride electrode inserted into it will be constant so too will be the potential
between hydrochloric acid and the inner surface of the glass bulb. Hence the only
potential which can vary is that existing between the outer surface of the glass bulb and
the test solution in which it is immersed, and so the overall potential of the electrode is
governed by the hydrogen ion concentration of the test solution.
2.4. When a highly alkaline solution is tested a reverse of the equilibrium reaction mentioned
in step 2.3 takes place i.e. sodium ions from the solution will pass into the hydrated layer
in preference to hydrogen ions and consequently the measured e.m.f. (and hence pH) are
too low. This is the reason for alkaline error for soda lime glass electrodes. Using
lithium in place of sodium in the glass composition reduces alkaline error. Similarly
strong acid solutions reduces the activity of water in the solution and it affects the
hydrated layer of the electrode which is involved in the ion exchange reaction. But the
error in the measurements due to this is of a much smaller degree.
2.5. Dependent upon the nature of the glass used in the construction of membrane, individual
character of each electrode, age of an electrode etc, an asymmetry potential may exist.
This can be measured by keeping a test solution identical with the internal hydrochloric
acid solution. Then the electrode will show a small potential which is found to vary with
time. In order to over come the asymmetry potential the electrode must be standardised
frequently by solutions of known hydrogen ion activity (buffer solutions).

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2.6. The temperature probe connected measures the sample temperature and corrects the pH
value for 25 0C.
3. APPARATUS
3.1. Combined pH glass electrode temperature sensor and meter WTW pMX 2000.
3.2. Magnetic stirrer.
4. REAGENTS
4.1. Buffers Technical.

Take 4.01, 7.00 and 10.00 buffer solutions from the readymade supply.
5. CALIBRATION
Connect the pH electrode to the ‘electrode’ socket and temperature sensor to the
‘TP’ socket and immerse the electrode in demin water.
5.1. Switch on the instrument and wait to finish the auto test.
5.2. Press <CAL> on the meter.
5.3. Select technical buffers from the meter. (Buffer 4.01, 7.00 & 10.00) by pressing the
<RUN> button.
5.4. Select any two buffers from the list and immerse the electrode into the first one and
shake gently.
5.5. Press <BUFFER ENTER> and press <RUN>.
5.6. Press <AUTO READ>.
5.7. When the buffer 2 is requested keep the second buffer and press <RUN>.
5.8. Now the slope will be displayed. If the slope displayed is not between -50 and -60 repeat
the calibration, press <RUN>.
5.9. If the next display of asymmetric potential millivolt is not between +45 & -45 repeat the
calibration. Now the instrument is ready for measurements.
6. PROCEDURE
6.1. Rinse the electrode with the sample and keep the sample.
6.2. Press <RUN>.
6.3. Press <pH.01> for 2 decimal reading and <pH.001> for 3 decimal reading.
6.4. Press <AUTO READ> for automatic measurement and Press <CONT> for continuous
measurement.
7. CALCULATION
pH can be directly read from the display.
Reference: pH/ION meter manual.

Page 159
C. DETERMINATION OF FLUORIDE (F -)
1. SCOPE
This electrode method covers an accurate measurement of fluoride at a concentration range

between .02 mg/L to saturated fluoride. This test method is applicable to the measurement of
fluoride ion in finished water, natural water and most industrial waste waters.
Simple and complex fluoride ions are found in natural waters. Fluoride complexes of aluminium,
silicon and boron are of industrial origin.
Fluoridation of drinking water to prevent dental caries is practiced by a large number of
communities, fluoride is monitored in drinking water to assure the quality of the water supplied
for (level 1.4 to 2.4 mg/L) human consumption.
2. PRINCIPLE
2.1. The fluoride ion is determined potentiometrically using an ion selective fluoride
electrode in conjunction with a standard single junction, sleeve type reference electrode,
and a pH meter having an expanded millivolt scale, or a specific ion meter having a
direct concentration scale for fluoride. The fluoride electrode consists of lanthanam
fluoride crystals across which a potential is developed by the fluoride ions.
2.2. Interferences
Extremes of pH interferes. Sample pH should be between 5 ~7 for lower ranges and
others may be between 5 ~ 9. Sample with >10,000 mg/l TDS is not suitable for testing
by this method. Polyvalent cations of Si4+,Fe3+, and Al3+ interferes by forming complexes
with fluoride. The degree of interference depends upon the concentration of fluoride and
pH of the solution. In solutions with a pH below 5, hydrogen ions complexes a portion
of the fluoride ions by forming HF or HF2 Which cannot be detected by the electrode.
To avoid this, sample pH is raised with sodium acetate buffers (do not use caustic, this
will distrub the total ionic strength). To avoid interferences, calibration standards may be
prepared in fluoride free back ground water.
3. APPARATUS
3.1. pH meter WTW pMX 2000 with expanded millivolt scale.

3.2. Fluoride ion selective electrode, reference electrode single junction sleeve type and
temperature sensor.
3.3. Magnetic mixer with teflon coated stirrer.
3.4. Pipettes, beakers etc.
4. REAGENTS
4.1. TISAB (Total Ionic Strength Adjusting Buffer)

4.2. Fluoride standards.
Sodium fluoride solution (1.0 ml = .01 mg F-).
Dissolve 0.2210 g of NaF in water and dilute to 1 litre. Dilute 100 ml of this solution
and dilute to 1 litre with DM water and store in a borosilicate glass or polyethylene
bottle.
4.3. 15% Sodium acetate.
Dissolve 150 g of CH3COONa in 1000 ml DM water.
5. CALIBRATION
5.1. Electrode preparation

5.1.1. Fix the temperature sensor and reference electrode into its respective sockets.
5.1.2. Check the reference electrode for internal solutions. Make up with respective
Page 160
filling solution (provided by the manufacturer) if the level is less.

5.1.3. Remove the protective cap of the fluoride electrode, fix it to the electrode
socket and it is ready for use.
5.2. Press <PROG> on WTW pMX 2000 meter.
5.3. Press <PROG NR> on the extended key board.
5.4. Press 6 and <ENTER>.
5.5. Press <AUTO> and <ENTER>
5.6. Press <g/l> and <ENTER>.
5.7. Select 2 standards covering the sample concentration. Prepare the standard by pipetting
suitable volume from the standard stock solution and make up the volume using demin
water or fluoride free back ground solution.
5.8. Pipette 40 ml of any one of the two standards into a beaker and mix with
20 ml of F- TISAB and immerse the electrode into it and keep on a slow stirring.
5.9. Enter the concentration in g/l of the present standard kept and press <ENTER>.
5.10. Press <RUN>. Now the meter will accept a stable standard value. At the end a second
standard is requested.
5.11. Keep 40 ml of the next standard, add 20 ml of F- TISAB into it and keep on slow
stirring.
5.13. Press <RUN>. The meter will again accept a stable value. At the end a display of the
slope will be visible on the screen. If the value is not ~56 mV repeat the calibration.
5.14. Press <RUN> again. Now the meter is ready for measurement
6. PROCEDURE
6.1. Press <PROG NR>, enter 12 and press <ENTER>.

6.2. Enter the dilution/concentration factor for the sample, if any, and press <ENTER>.
6.3. Pipette 40 ml sample and 20 ml F- TISAB into a beaker, mix by stirring and immerse the
electrode into it.
6.4. Press <AUTO> and <RUN>. After attaining a stable reading the concentration is
displayed on the screen.
6.5. Electrode storage
Reference electrode: After the use wash and keep in demin water and for more than one
week store dry.
Fluoride electrode: After the use wash the electrode with demin water, blot dry, fix the
protective cap and store dry.
6.6 If a concentrated acid is to be tested
Prepare standards also in the same acid matrix. In place of ready made TISAB, 15%
sodium acetate is used. Sample and standards are diluted in the ratio of 9:1 (9 parts
sodium acetate and 1 parts of sample or standard) for calibration and measurement.
6.6.1. Follow the calibration procedure with the standard and sodium acetate
mixture.
6.6.1 For sample measurement pipette 5 ml of the sample and 45 ml of sodium
acetate and and continue steps 6.4.
7. CALCULATION
The concentration is directly read from the display.

References:
1) pH/ION meter manual.
2) ASTM D 1179
3) Literature from The Weir Westgarth Ltd.

Page 161
D. DETERMINATION OF NITRATE (N03-)

1. SCOPE
Nitrates are generally present in the sewage effluent water. The level of nitrates present gives an
idea of the oxidation level of the organic waste present in the treated water. Nitrate presence in
the drinking water may be suspected to be possible fecal contamination of the water. Nitrates
stimulates algal and aquatic growth. Nitrate presence more than 0.3 ppm in drinking water
causes disease methemoglobinemia in human infants (blue babies).
2. PRINCIPLE
2.1. The nitrate ion electrode is a selective sensor that develops a potential across a thin
porous inert membrane made up of Tris-(1—10 phenanthroline) Nickel II nitrate, that
holds in place a water immiscible liquid ion exchanger. The electrode responds to NO3
ion activity between about 0.4 to 62,000 mg/L of N03.
2.2. Interference
F- 600 C032- 20 SO42- 1000 CN- 0.1
Cl- 30 HCO3- 10 PO4- 50 OAC 200
Br- 0.7 HPO42- 50 NO2- 0.7 ClO3- 0.05
I- .005 H2P04- 50 HS- 1 ClO42- .0001
Any of the components exceeding the above concentration in ppm will cause
interference.
3. APPARATUS
3.1. pH/ion meter WTW pMX2000 with expanded millivolt scale.
3.2. Double junction reference electrode. Fill the outer chamber with (NH4)2S04
solution
3.3. Nitrtate ion electrode.
3.4. Magnetic stirrer (teflon coated).
4. REAGENTS
4.1. Nitrate—free demin water.
4.2. Stock nitrate solution.
Dry potassium nitrate (KNO3) in an oven at 105 °C for 24 hours. Dissolve 0.16306
grams in demin water and dilute to 1000 ml (1.0 ml 100 pg N03). Preserve with 2 ml
of CHC13/L. This solution is stable for 6 months.
4.3. Alternately standard solutions also can be prepared from the ready made stock solution.
4.5. Standard nitrate solutions. Dilute 1.0, 10.0 and 50 ml of stock nitrate solution to 100 ml
with demin water to obtain standard solutions of 1.0, 10 and 50 mg NO3 /L respectively.
4.6. Buffer solution (TISAB).
a).Dissolve1.732grams of Al2(SO4) 3.18H20, 3.43grams of Ag2SO4, 1.28 grams of
H3BO3 and 2.52grams of sulfamic acid (H2NSO3H) in about 800 ml water.
Adjust to pH 3.0 by slowly adding
0.10N NaOH. Dilute to 1000 ml and store in a dark glass bottle.
b).Ready made buffer solution (NO3 TISAB) available also can be used.
4.7. Sodium hydroxide 0.10 N.
4.8. Reference electrode filling solution.
Dissolve 0.53 grams of (NH4)2S04 in demin water and dilute to 100 ml.

Page 162
5. CALIBRATION
filling solution (provided by the manufacturers) if the level is less. Fill the
outer chamber with (NH4)2S04 solution.
5.1.3. Screw on the measuring head of the nitrate electrode with sealing ring. Shake
the electrode like a thermometer and fix the electrode with the
‘electrode’ socket and keep immersed in demin water for 15 minutes.
Then immerse the electrode in standard nitrate solution (0.10 ppm) for
one hour. After one hour wash the electrode in demin water and it is
ready for measurements.
5.5. Press <AUTO> and <ENTER>.
5.7. Select any of the two standards from the steps 4.5 (covering the sample concentration
range). Standards are to be selected keeping a minimum concentration difference of one
decade between standards.
5.8. Take 50 ml of the first standard and add 1 ml of NO3 TISAB into it and stir slowly for
1 minute.
5.9. Enter the concentration in g/1 of the present standard kept and press <ENTER>.
5.10. Press <RUN>.
5.11. Keep 50 ml of the second standard and add 1 ml of N03 TISAB stir slowly
for 1 minute.
5.13. Press <RUN>, now the meter will accept a stable value and the slop will be
displayed. The expected slope is ~56 mV.
5.14. Press <RUN>. again and the meter is now ready for measurements.
6. PROCEDURE

6.2. Enter the dilution/concentration factor of the sample, if any, and press <ENTER>.
6.3. Pipette 50 ml of sample and 1 ml of N03 TISAB and keep on slow stirring.
6.4. Press <AUTO> and <RUN>.
After attaining a stable reading the concentration is displayed on the display.
6.5. Storage of electrodes
Nitrate electrode: After the use keep in dilute nitrate solution (0.10 ppm) and for more
than two days wash the electrode in demin water, blot dry, remove the measuring head
and store dry.
Reference electrode: After the use keep in demin water and for more than one week
store dry.
7. CALCULATION
Read the concentration directly from the display.
References: 1) pH/ion meter manual.

2) APHA 4500 — N03 - D.

Page 163
E. DETERMINATION OF POTASSIUM (K+)

1. SCOPE
Potassium ranks seventh among the elements in order of abundance, yet its concentration in
most drinking waters seldom reaches 20 mg/L.
2. PRINCIPLE
2.1. Potassium ion selective electrode is having a membrane made up of the antibiotic
valinomycin. The electrode is set up with an internal potassium chloride solution of
definite concentration from which some potassium ions are extracted by the valinomycin
into the inner surface of the membrane. When the electrode is placed in an aqueous
solution containing potassium ions some of these are extracted into the external surface
of the membrane and being the internal concentration constant the resultant
electrode potential will be dependent upon the potassium ion activity of the test solution.
The electrode responds to potassium ion concentration between 0.4 to 39,000 mg/L.
2.2. Interferences
10 % Error in the case of thefollowing concentration ratio (interference ion /
measurement ion).
H+ 10 Ag+ 1000 NH4+ 6

Li+ 2000 Cs+ 0.3 Tris+ 1000
Na+ 2000 Tl 6
3. APPARATUS
3.1. pH/ion meter with expanded millivolt scale.

3.2. Double junction reference electrode. Fill outer chamber with (NH4)2S04 solution.
3.3. Potassium ion electrode.
4. REAGENTS
4.1. Stock potassium solution.

Dissolve 0.1907 gram of potassium chloride KC1 (dried at 110 °C) in demin water and
make up to 1000 ml (1.0 ml 100 = µg K)
4.2. Alternately standard solutions also can be prepared from the ready made stock solution.
4.3. Standard potassium solutions. Dilute 1.0, 10.0 and 50 ml of stock potassium solution
100 ml with demin water to obtain standard solutions of 1.0, 10.0 and 50 mg/L
respectively.
4.4. Buffer solution (TISAB) - 6 mol/L of sodium chloride NaCl. Dissolve 35.065 gram of
NaC1 in demin water and dilute to 100 ml (l ml =138 mg Na.)
4.5. Reference electrode filling solution.
Dissolve 0.53 gram of (NH4)2S04 in demin water and dilute to 100 ml.
5. CALIBRATION
5.1 Electrode preparation

filling solution (provided by the manufacturers) if the internal level is less.
Fill the outer chamber with (NH4)2S04 solution.
5.1.3. Screw on the measuring head of the potassium electrode with sealing ring.
Shake the electrode like a thermometer and f ix the electrode with the
Page 164
‘electrode’ socket and keep immersed in demin water for 15 minutes. Then
immerse the electrode in standard potassium solution (1 ppm) for one hour.
After one hour wash the electrode in demin water and it is ready for
measurements.
5.8. Take 100 ml of the first standard and add 1 ml of potassium TISAB into it and stir
slowly for 1 minute.
5.10. Press <RUN>.
5.11. Keep 100 ml of the second standard and add 1 ml of potassium TISAB and stir slowly
for 1 minute.
5.13. Press <RUN>, now the meter will accept a stable value and the slop will be displayed.
The expected slope is ~56 mV.
5.14. Press <RUN>. again and the meter is now ready for measurements.
6. PROCEDURE
6.1 Sample preparation

Sodium interferes when the concentration of sodium in the measuring media exceeds
more than 2000 times of measuring ion (potassium) concentration. Since TISAB added
is 6 molar NaC1 the sample chloride (assuming that all chlorides are sodium chloride)
level is to be brought to <500 ppm by diluting with demin water. After dilution the
potassium concentration must be more than 1 ppm for a chloride concentration of <500
ppm in the 100 ml of the test solution
6.4. Pipette 100 ml of sample and 1 ml of potassium TISAB and keep on slow stirring.
After attaining a stable reading the concentration is displayed.
6.6. Cleaning of potassium electrode
Put sluggish and drifting electrodes in distilled water for one hour, then keep in dilute
standard solution for several hours.
Potassium electrode: After the use keep in dilute potassium solution (1 ppm) and for
more than two days wash the electrode in demin water, blot dry, unscrew from the main
body and store dry.
store dry.
7. CALCULATION

References: 1) pH/ion meter manual.
2) APHA
3) VOGEL

Page 165
F. DETERMINATION OF CALCIUM (Ca++)
1. SCOPE
The calcium is fifth among the elements in order of abundance. Small concentrations of calcium
carbonate combat corrosion of pipe lines by laying down a protective coating. Appreciable
calcium salts on the other hand precipitate on heating to form harmful scale in boilers and
cooking utensils. Calcium gives good taste to drinking water as well as it is very much essential
for human body. Calcium contributes to the total hardness of water.
2. PRINCIPLE
2.1. The calcium electrode is a selective sensor for calcium ions.. The membrane of the electrode
is made up of o—nitrophenyl octyl ether prepared in a P V C matrix. A solution of a
specified calcium concentration filled inside creates a definite potential on the inside surface
of the membrane.On the outer side of the membrane the potential established will be
determined by the calcium ion activity of the test solution.
2.2. Interference
10 % Error in the case of the following concentration ratio (interference ion/ measurement
ion)
H+ 4 NH4’ 200 Sr2+ 6 Fe2’ 20
+ 2+ 2+
Li 300 Pb 0.01 Hg 4 Cu2’ 40
Na+ 200 Ni2+ 50 Zn2+ 1000 Tris+ 300
+ 2+ 2+
K 400 Ba 700 Mg 1000
pH of the measuring water must be between 2.5 and 11.0 at a temperature

between 0 — 40 0C.
3. APPARATUS
3.1. pH/ ion meter WTW pMX 2000 with expanded millivolt scale.
3.2. Double junction reference electrode.
3.3. Calcium ion selective electrode.
4. REAGENTS
4.1. Calcium stock solution.
Suspend 0.2497 gram of CaCO3 (dried at 180 °C for 1 hour before weighing ) in water
and dissolve cautiously with a minimum amount of 1+1 HNO3. Add 10.0 ml of con
HNO3 excess and make up to 1000 ml with demin water (1.0 ml = 100 µg Ca)
4.2. Alternately standard solution also can be prepared from ready made stock solution.
4.3. Standard calcium solution. Dilute 1.0, 10.0 and 50 ml of the stock calcium solution to
100 ml with demin water to obtain standard solution of 1.0,
10.0 and 50 mg Ca/L respectively.
4.4. Reference electrode filling solutions. Supplied by the manufacturer.
5. CALIBRATION
filling solutions (provided by the manufacturers) if any of the levels are less.
5.1.3. Screw on the measuring head of the calcium electrode with sealing ring.
Shake the electrode like a thermometer and fix the electrode with the

Page 166
‘electrode’ socket and keep immersed in demin water for 15 minutes. Then
immerse the electrode in standard calcium solution (10 ppm) for one hour.
After one hour wash the electrode in demin water and it is ready for
measurements.
5.5. Press <AUTO> and (ENTER>.
5.8. Take 50 ml of the first standard and add 1 ml of calcium TISAB into it and stir slowly
for 1 minute.
5.10. Press <RUN>.
5.11. Keep 50 ml of the second standard and add 1 ml of calcium TISAB and stir slowly for
1 minute.
5.14. Press <RUN> again and the meter is now ready for measurements.
6. PROCEDURE
6.1. Sample preparation
Dilute the samples if any of the interfering radicals exceed the given limits
6.3. Enter the dilution/concentration factor of the sample, if any, and press
<ENTER>.
6.4. Pipette 50 ml of sample and 1 ml of calcium TISAB and keep on slow stirring.
6.6. Cleaning of calcium electrode
Put sluggish and drifting electrodes in distilled water for one hour, then keep in dilute
standard solution for several hours.
Calcium electrode: After the use keep in dilute calcium solution (10 ppm) and for more
than two days wash the electrode in demin water, blot dry, unscrew from the main body
and store dry.
store dry.
7. CALCULATION
References:
1) pH/ion meter manual.
2) APHA
3) VOGEL

Page 167
C. DETERMINATION OF SODIUM (Na+)
1. SCOPE
Sodium ranks sixth among the elements in order of abundance and is present in most natural
waters. The ratio of sodium to total cations is important in agriculture and human pathology. Soil
permeability can be harmed by high sodium ratio, Sodium is not particularly significant in
potable water except for those persons having an abnormal sodium metabolism.
2. PRINCIPLE
2.1. The sodium ion electrode is made up of special glass containing aluminium oxide. Like
all potentiometric electrode measuring systems this is responsive to changes in ion
activity and not true concentration changes (that is the response is to changes in
concentration multiplied by an activity coefficient). However, as concentrations
approach infinite dilution, activity coefficients approach unity and ion concentration and
active ion concentration become very nearly equal.
2.2. The activity coefficient of the sodium ion will vary with changes in the total ionic
strength of the solution. There for it is important to maintain either a low or a constant
ionic strength.
2.3. The signal from the sodium ion electrode in a very dilute solution varies approximately
59-mV/ decade change in Na+ concentration at 25 °C.
2.4, This test method allows measurement of sodium in ppb level for monitoring condensate
and all low sodium water sources.
2.5. Interferences
1 % error in the case of the following concentration ratio (interference ion/ measurement
ion).
H+ 10-3 K+ 5 NH4+ 0.2 Li+ 1 Ag+ 0
3. APPARATUS
3.1. pH / ion meter WTW pMX 2000 with expanded millivolt scale.
3.2. Sodium ion selective electrode.
3.4. Double junction reference electrode.
4. REAGENTS
4.1. Stock sodium solution. (1.0 ml = 100 µg Na).
Dissolve 0.25418 gram of NaC1 (dried to a constant weight at 105 °C) and dilute to 1
litre in a polythene bottle (alkali free).
4.2. Alternately standard solution also can be prepared from ready made stock solution.
4.3. Sodium standard solutions.
Pipette 1.0, 10.0 and 50 ml of the above stock solution and dilute to 100 ml. This
solution will be equal to 1.0, 10.0 and 50 mg Na/L respectively.
4.4. Buffer solution (TISAB).
Ready made buffer solution containing morpholine available can be used.
5. CALIBRATION

filling solutions (provided by the manufacturers) if any of the levels are less.

Page 168
5.1.3. Soak the electrode in 0.1 molar NaC1 solution for several hours if the
electrode membrane is dried out. Rinse the electrode in demin water and it is
ready for measurements.
5.2. Press <P1100> on WTW pMX 2000 meter.
5.3. Press <PROG NR) on the extended key board.
5.8. Take 20 ml of the first standard and add 10 ml of sodium TISAB into it and stir slowly
for 1 minute.
5.10. Press <RUN>.
5.11. Keep 20 ml of the second standard and add 10 ml of sodium TISAB and stirr slowly for
1 minute.
5.14. Press <RUN> again and the meter is now ready for measurements.
6. PROCEDURE
6.1. Sample preparation

Dilute the samples if any of the interfering radicals exceed the given limits
6.4. Pipette 20 ml of sample and 10 ml of sodium TISAB and keep on slow stirring.
6.6. Cleaning of sodium electrode
6.6.1. Mild contamination:Clean with mild soap and water.
6.6.2. Heavy contamination: Dissolve the top layer by putting in approx.
2.5 mol/L NaOH (30 minutes). Treatment in hydrofluoric acid may be
considered as a last resort (a few minutes).
6.6.3. After each cleaning procedure put the electrode in sodium chloride solution
(0.1 mol/L) for several hours.
Sodium electrode: After the use store dry with the protective cap. Reference electrode:
After the use keep in demin water and for more than one week store dry.
7. CALCULATION
References:
2) ASTM D 4191, 3561 and 2791.
3) VOGEL

Page 169
H. DETERMINATION OF AMMONIA (NH3)
1. SCOPE
Nitrogen is a nutrient in the environment and is necessary to sustain growth of most organisms.
It exists in several forms such as nitrate, nitrite, organic, and ammonia.
Ammonia is a colourless, gaseous compound with a sharp distinctive odour. It is highly soluble
in water where it exists in a molecular form associated with water and in an ionized form as
NH4+. The extend of association or ionization is dependent on the temperature and pH. It may
also he toxic to aquatic life. The extend of toxicity is dependent upon species and extent of
dissociation. Arnmonia. may occur in water as a product of anaerobic decomposition of
nitrogen containing compound or from waste streams containing ammonia.
This method is applicable to the measurement of 0. 05 to 850 mgNH3 – N/L in river water,
saline water, and waste water.
2. PRINCIPLE
2.1 The sample is made alkaline with sodium hydroxide to convert ammonium ion to
ammonia. The ammonia thus formed diffuses through a gas permeable
membrane of a selective - ion electrode and alter the pH of its internal solution
which, in turn, is sensed by a pH electrode. The potential is measured by means of
a specific – ion
meter.The ammonia content is read directly from the meter after calibration.
2.2 Interferences
Volatile amines are positive interferences.
Mercury and silver interfere by complexing with ammonia.
3. APPARUTUS
3.1 pH /ion meter WTW pMX 2000 with Extender keydoard.

3.2 Ammonia ion selective electrode – WTW NH 500/2.
3.3 Temperature sensor – WTW FTK 530.
3.4 Desk stand with electrode holder.
3.5 Magnetic stirrer.
3.6 Beaker, 150 ml.
3.7 Measuring cylinder, 100 ml.
3.8 Volumetric flasks , 1L ,100 ml.
3.9 Tensette pipette, 10 ml, 1 ml.
4. REAGENTS.
4.1 Alkaline reagent - NaOH mol / L

WTW Model MZ/NH3/CN – Cat.No.150 130
4.2 Ammonia solution, Stock , 10g/L NH4+
WTW Model – ES/NH4 – cat.No. 120 240.
4.3 Ammonia standard solutions, Intermidiate.(1000, 100, 10 mg/L)
Pipette 10.0 ml of the above stock solution to a 100 ml volumetric flask and make to 100
ml with ammonoia free DM water. This solution will be equal to 100 mg NH4+ /L
Dilute the 1000mg/L standard solution 10 times to get 100 mg/L and 100 times to get 10
mg/L standard solutions.
4.4 Addition standard solution, 20 mg NH4+/L
Pipette 2.0 ml of 1000 mg/L standard to 100 ml volumetric flask and make upto the mark
with ammonia free DM water.
Page 170
4.5 Electrode storing solution.

Add 1% alkaline reagent solution in DM water.
5. CALIBRATION
5.1.1. Remove electrode protective cap.
5.1.2. Rinse electrode with deionized water.
5.1.3. Fill approximately 1 ml electrolytic solution into an exchange head.
5,1.4. Remove air bubbles in the electrolytic space by knocking.
5.1.5. Screw exchange head onto the meter with electrode.
5.1.6. Connect the electrode to the meter with cable.
5.2. Connect the temperature sensor to the TP socket.
5.3. Keep the electrode immersed in electrode storing solution for 1 hour for stabilization.
5,5. Press <PROG NR> on the extender keyboard.
5.6. Enter 9 and press <ENTER>.
5.7. Press <MAN> and <ENTER>.
5.8. select any of the two standards covering the sample concentration range. Standards are
to be selected keeping a minimum concentration difference of one decade between
standards.
Note: Select two standard as follows:
For sea water and outfall STD 1 = 0.05 mg/L NH4+
STD 2 = 0.5 mg/L NH4+
For sewage sample STD 1 = 0.1 mg/L NH4+
(Range 0.5 to 1.0 mg/h) STD 2 = 1.0 mg/L NH4+
For sewage sample STD 1 = 1.0 mg/L NH4+
(Range 1.0 to 10 mg/L) STD 2 = 10 mg/L NH4+
For sample of high conc STD 1 = 10 mg/L NH4+
(Range 10 to 900 mg/L) STD 2 = 100 mg/L NH4+
5.9. Take 100 ml. of the lower concentration standard in a 150 ml beaker.
5.10. Add 1 ml of alkaline reagent and put the stirring rod in it.
5.11. Keep the beaker on the magnetic stirrer and keep the stirring ON with a medium speed.
5.12. Clean the electrode and the temperature probe with PM water and wipe with clean tissue
paper.
5.13. Immerse the electrode and TFK 530 into the solution and press <RUN>.
5.14. Wait for stable reading
(Note: 30 minutes for standards of concentration <1 mg/L and 5 minutes for standards of
concentration > 10 mg/L and for the rest of standards give 15 minutes.
5.15. Press <ENTER>
5.16. Take 100 ml. of the higher concentration standard in a 150 ml beaker.
5,19. Clean the electrode and the temperature probe with PM water and wipe with clean tissue
paper.
5.21. Wait for stable reading
(Note: 30 minutes for standards of concentration <1 mg/l and 5 minutes for standards of
concentration > 10 mg/l and for the rest of standards give 15 minutes.
5.22. Press <ENTER>
5.23. Note the slope displayed in mV.
5.24. Press <RUN> and now the meter is ready for measurement.

Page 171
6. PROCEDURE

6.1.1. Remove electrode protective cap.
6.1.2. Rinse electrode with deionized water.
6.1.3. Fill approximately 1 ml electrolytic solution into an exchange head.
6.1.4. Remove air bubbles in the electrolytic space by knocking.
6.1.5. Screw exchange head onto the meter with electrode.
6.1.6. Connect the electrode to the meter with cable.
6.2. Connect the temperature sensor to the PP socket.(No reference electrode)
6.3. Keep the electrode immersed in electrode storing solution for 1 hour for stabilization.
6.5. Press <PROG NR> on the extender keyboard.
6.6. Enter 13 and Press <ENTER>.
6.7. Press <MAN> and <ENTER>.
6.8. Press <g/l> and <ENTER>
6.9. Enter the slope reading and press <ENTER>.
Note: Approximate slope reading are as follows:
<0.5 mg/L range = - 38 mV
0.5 to 1.0 mg/L range = - 44 mV
1.0 to 2.0 mg/L range = - 51 mV
> 2.0 mg/L range = - 59 mV
6.10. Enter the addition proportion in % (P)as 1 and press <ENTER>.
6.11. Enter the Factor (F) as 0.954 and press<ENTER>.
6.12. Enter the addition standard concentration as 0.02 and press <ENTER>.
6.13. Take 100 ml. of the sample in a 150 ml beaker.
6.16. Clean the electrode and the temperature probe with PM water and wipe with clean tissue
paper.
6.18. Wait for 30 minutes for stable reading.
Note: For samples of higher ammonia concentration 5 — 10 minutes is sufficient.
6.19. Press <ENTER>
6.20. Add 1 ml addition standard (20 mg/L NH4+) into the sample and press <RUN>.
6.21. Wait for stable reading.
6.22. Note the concentration displayed on the meter as ammonia content in the sample as
mg NH3—N/L
6.23. After the analysis store the electrode in electrode storing solution.
Note: For storage of more than 2 days, remove the exchange head and rinse with
PM
water and screw on the electrode protection cap.
7. CALCULATION
Ammonia as NH3 - N = Displayed reading in mg NH3—N/L
References:
2) Ammonia electrode NH 500/2 instruction manual
2) ASTM D 1426
3) APHA 4500 NH3 F&G

Page 172
H. DETERMINATION OF SULPHIDE (S2 -)

1. SCOPE
Sulphide ion is found in ground water and waste waters, causing odour and corrosion
problems. Its common presence in waste water comes partly from the decomposition of
organic matter, some times from industrial waste, hut mostly from the bacterial reduction
of sulphate. If acidified, waters can release hydrogen sulphide, which is extremely toxic
even at low levels. This method provides a means for interference free measurement of
free sulphide ion.
This test method is applicable for the measurement of 0.01 to 4000 mg/L Sulphide in
water.
Note: This standard procedure may involve hazardous material operations and
equipment. Use appropriate safety and health practices. Sulphide samples, when
acidified, can release highly toxic hydrogen sulphide gas.
2. PRINCIPLE
Sulphide ion is measured potentiometrically using a sulphide ion selective electrode in
conjunction with a double—junction sleeve type reference electrode. Potentials are read
using a specific ion meter having a direct concentration scale for sulphide ion.
Samples are treated prior to analysis with sulphide anti-oxidation buffer (SAOB II). This
buffer fixes the solution pH at highly alkaline level and contains ascorbic acid to retard air
oxidation of sulphide ion in solution.
3. APPARATUS
3.1. pH/ ion meter WTW pMX 2000 witb Fxtender- Keyboard,
3.2. Sulphide/Silver ion selective electrode — WTW Ag 500.
3.3. Reference electrode — WTW R 502
3.4. Desk stand with electrode holder.
3.6. Plastic beaker, 120 ml.
3.8. Volumetric flasks, lL , 100 ml.
3.9. TenSette pipette, 10 ml, 1 ml.
3.10 Analytical balance
4. REAGENTS
4.1. Silver standard solution, 1 g/L
Weigh accurately 1.5748 g of Silver nitrate (AgNO3) and dissolve in de-ionized
water. Transfer qualitatively to a 1 litre volumetric flask and make up to the mark
with de-ionized water.
4.2. Sulphide Anti-Oxidant Buffer (SAOB II)
In a 1000 ml beaker containing approx. 600 ml of de-ionized water, add 200 ml of
10 M sodium hydroxide (or 80 g pellets), 35 g of ascorbic acid, and 67 g of
disodium EDTA. Stir until everything dissolves and transfer the solution to a 1000
ml volumetric flask. Dilute to the mark with de-ionized water.
Note: Freshly prepared SAOB is colourless or slightly brownish yellow.
If the solution has strongly oxidized, its colour turns towards dark
brown. In this case the solution must not he used any more.
4.3. Sodium sulphide Solution, Stock, (~10 g/L S)
Weigh 7.5 g of sodium sulphide 9-hydrate (Na2S. 9H20) in a beaker and add 50 ml
SAOB solution to it and mix to dissolve. Make up to 100 ml with de-ionized
water.
Page 173
Note: Since Sodium sulphide is available with varying water content, concentration
determination is necessary, for its use as calibration standard.
4.4. Nitric acid

Concentrated nitric acid (65%)
4.5. Sample conditioning solution for silver WTW model ISA/FK Cat.No.140 110
5. CALIBRATION
5.1. Concentration determination of sulphide stock solution.
5.1.1. Preparation of calibration standard, Ag+ = 50 mg/L.
Pipette 5.0 ml of the standard solution Ag+ = 1g/L to a 100 ml volumetric
flask. Add 2 ml sample conditioning solution ISA/FK and 2 ml nitric acid
(65%) to it and make up to the mark with de-ionized water. Mix well and
pour into the 120 ml plastic beaker.
5.1.2. Preparation of calibration standard, Ag+ = 500 mg/L.
Pipette 50 ml of the standard solution Ag+ = lg/L to a 100 ml volumetric
5.1.3. Preparation of standard sample, Ag+ = 950 mg/L.
Pipette 95 ml of the standard solution Ag+ = 1g/L to a 100 ml volumetric
5.1.4. Rinse electrode with deionized water. Keep the electrode and the reference
electrode immersed in a blank solution containing 2 ml each of nitric acid and
ISA/FKand made to 100 ml with deionized water.
5.1.5 Press <PROG> on WTW pMX 2000 meter
5.1.6. Press <PROGNR> on the extender keyboard.
5.1.7. Enter 6 and press <ENTER>.
5.1.8. Press <AUTO> and <ENTER>,
5.1.9. Press<g/l>and press <ENTER>
5.1.10. Enter the concentration in g/l of the lower standard (0.050) and press
<ENTER>
5.1.11. Blot dry the clean electrodes and immerse them in the lower standard solution
kept in the plastic beaker and keep the stirrer off after 1 minute running and
press<RUN>.
5.1.12. The meter will accept a stable reading and will request for the second
standard concentration.
5.1.13. Enter the concentration in g/l of the second higher standard (0.500) and press
<ENTER>
5.1.14. Clean the electrode with de-ionized water and immerse the electrodes in the
second higher standard solution kept in the plastic beaker and keep the stirrer
off after 1minute running and press <RUN>.
5.1.15. The meter will accept a stable reading and will display the slope reading (~59
mV).
5.1.16. Press <RUN> to complete the step.
5.1.17. Clean the electrode with deonized water arid keep it immersed in blank water.
5.1.18. Press <PROGNR> on the extender keyboard.
5.1.19. Enter 12 and press <ENTER>
5.1.20. Enter the factor F as 1.0 and press <ENTER>
5.1.21. Blot dry the electrodes and immerse them in the standard sample solution
(Ag+ = 950 mg/L) and keep the stirrer running for 1 minute and note the
Page 174
concentration displayed as the concentration of the standard sample solution.

5.1.22. Press <PROG NR> on the extender keyboard.
5.1.24. Press <MAN> and <ENTER>.
5.1.25. Press (g/l) and press <ENTER>
5.1.26. Enter slope reading by pressing <ENTER> when the slope is displayed.
5.1.27. Enter the sample addition volume P% as 0.5 and press <ENTER>
5.1.28. Enter the factor F as 0.1485 and press <ENTER>.
5,1.29. Enter the concentration of the standard sample solution (~0.950) and press
<ENTER>
5.1.30. Blot dry the electrode and immerse the electrode in the standard sample
solution
(950 mg/L). Press <RUN> when sample 1 is displayed.
5.1.31. When the reading is stable press <ENTER>
5.1.32. Add 0.5 ml of the sodium sulphide stock solution when sample 2 is requested.
5.1.33. Keep the stirrer running and press <RUN>
5.1.34. When the reading is stable, note the concentration of the sodium sulphide in
g/L
5.2 Electrode calibration for sulphide determination.
5.2.1. Diluted sulphide standard stock solution (~0.10 g/L), Pipette 1.0 ml of the
standardized sulphide stock solution.
(~10 g/L) to a 100 ml volumetric flask and add 50 ml SAOB II solution and
make up to the mark with dc-ionized water.
5.2.2. Take 50 ml SAGE II solution each in two 100 ml volumetric flasks and add 0.1
ml and 1.0 ml of the diluted standard sulphide solution, respectively, and make
up to the marks with de-ionized water.
5.2.3. Transfer the prepared solutions to the plastic beakers.
5.2.4. Prepare a blank by mixing 50 ml de-ionized water with 50 ml SAOB.
5.2.5. Clean the electrode and keep them immersed in the blank solution.
5.2.6. Press <PROG NR> on the extender keyboard.
5.2.8. Press <AUTO> and <ENTER>.
5.2.9. Press <g/l> and press <ENTER>
5.2.10. Enter the concentration in g/l of the lower standard (1/10000 of the diluted
sulphide standard stock solution concentration) and press (ENTER>
5.2.11. Immerse the electrodes in the lower standard solution kept in the plastic beaker
and keep the stirrer running and press <RUN>.
5.2.12. The meter will accept a stable reading and will request for the second standard
concentration.
5.2.13. Enter the concentration in g/1 of the second higher standard (1/1000 of the
diluted sulphide standard stock solution concentration) and press <ENTER>
5.2.14. Clean the electrode with de-ionized water and immerse the electrodes in the
second higher standard solution kept in the plastic beaker and keep the stirrer
running and press <RUN>.
5.2.15. The meter will accept a stable reading and will display the slope reading(~29
mV).
5.2.16. Press <RUN> to complete the step.
5.2.17. Clean the electrode with dc-ionized water and keep it immersed in blank water.
6. PROCEDURE
6.1. Connect the electrodes to the meter with cable.
6.2. Keep the electrode immersed in blank water (50 ml de-ionized water plus 50 m1SAOB
II) and keep the stirrer running.

Page 175

6.4. Press <PROG NR> on the extender keyboard.
6.5. Enter 12 and Press <ENTER>.
6.6. Enter the dilution/concentration factor for the sample, as 2 and press <ENTER>.
6.7. Pipette 50 ml sample and 50 ml SAGE II into the plastic beaker, mix by stirring and
allow to stand for 5 minutes.
6.8. Blot dry the electrode and immerse the electrode into the sample solution
6.9. After attaining a stable reading note the concentration displayed on the screen.
6.10. Between samples, rinse the electrode with water, blot dry, and immerse in a blank
solution of 50 ml SAOB II plus 50 ml de-ionized water.
7. CALCULATION
Sulphide as S2- = Displayed reading in the specified unit.
References:
2) Sulphide electrode Ag 500 instruction manual
2) ASTM D 4658
3) APHA 4500 S

Page 176
SECTION : WATER
THE DETERMINATION OF IRON
(Orthophenanthroline method)

Date issued : 22ndMarch 1995
Page 177

IRON (ORTHPHENANTHROLINE METHOD)
G-LAB METHOD No.43
(By K.P. Abraham)
1. SCOPE
This method is applicable to the determination of total, dissolved and ferrous iron in drinking
water, cooling water and waste water having iron concentration between 0.05 and 3 mg/litre.
2. PRINCIPLE
Iron is brought into solution, reduced to ferrous state by boiling with acid and hydroxylamine,
and treated with, 1,10 phenanthroline at pH 3.2 to 3.3. Three molecules of phenanthroline
chelate each atom of ferrous iron to form an orange—red complex. The coloured solution
obeys Beer’s law and the intensity is proportional to the iron concentration.
Interference: Strong oxidising agents, nitrates, phosphates, chromium and zinc in

concentrations exceeding 10 times that of iron and cobalt and copper in excess of 5 mg/l will
interfere in this method.
3. APPARATUS
JASCO Model V 530.
3.3. Acid-washed glassware
Wash all glassware with concentrated hydrochloric
acid (HC1) and rinse
with distilled water before use to remove deposits
of iron oxide.
3.3.1. Beakers, 250 ml, 10 Nos.
3.3.2. Volumetric flasks, 50 ml, 10 Nos.
3.3.3. Volumetric flasks, 100 ml and 1 litre.
3.3.4. Burette, 25 ml.
4. REAGENTS
Use reagents low in iron. Use iron free distilled water in preparing standards and reagent
solutions. Store reagents in glass stoppered bottles.
4.1. Hydrochloric acid (HC1 Sp. gr. 1.19)
Concentrated HC1, containing less than 0.00005% Fe.
4.2. Hydroxylamine solution
Dissolve 10 g hydroxylamine hydrochloride (NH2OH.HC1) in distilled water and
dilute to 100 ml.
4.3. Ammonium acetate buffer solution
Dissolve 250 g ammonium acetate (NH4C2H3O2) in 150 ml distilled water. Add 700
ml concentrated (glacial) acetic acid. Because, even a good grade of ammonium
acetate contains a significant amount of iron, prepare new reference standards with
each buffer preparation.
4.4. Sodium acetate solution
Dissolve 200 g sodium acetate (NaC2H3O2.3H20) in 800 ml distilled water.
4.5. Phenanthroline solution
Dissolve 100 mg of 1,10 — phenanthroline monohydrate (C12H8N2.H20) in 100 ml
GLAB-43 (Revised) - Iron (Phenanthroline method) Page 2 of 3
Page 178
distilled water by stirring and heating to 80C. Do not boil. Discard the solution if it
darkens. Heating is unnecessary if 2 drops con. HC1 are added to the distilled water.
4.6. Stock iron solution, standard (1ml = 1mg Fe)
Spectroscopic standard as purchased (lml = 1 mg Fe) or prepare as follows:
Use electrolytic iron wire or iron wire for standardising” to prepare the
solution. Weigh 1.000 g wire and place in a 1000 ml volumetric flask.
Dissolve in 100 ml 6N sulphuric acid (H2SO4) and dilute to the mark.
4.7. Standard iron solution (lml = 0.01 g Fe)
Pipette 10.0 ml of the iron stock solution (lml = 1mg) to l000ml volumetric flask and
make upto the mark.
5. CALIBRATION
Take the standard iron solution (1 ml = 0.01 mg Fe) in the burette and transfer 2.5, 5.0, 7.5,
10.0, 12.5 and 15.0 ml of the standard iron solution (lml = 0.01 mg Fe) to clean 250 ml acid
cleaned beakers and make up the volume to about 50 ml with distilled water. Take 50 ml
distilled water as blank in another beaker. Add 2ml concentrated HC1 hydrochloric acid and
lml hydroxylamine hydrochloride reagent. Heat to reduce volume to 20 to 30 ml. Cool it and
add 10 ml ammonium acetate buffer. Add 2m1 phenanthroline reagent. Transfer to 50ml
volumetric flasks and make up to the mark. Wait for 10 minutes. Measure the absorbance
against the distilled water reagent blank at 510nm wavelength using 10mm cells. Plot a
calibration graph for concentration (mg/litre Fe) virsus absorbance. The above prepared
solutions are equivalent to 0.5, 1.0, 1.5, 2.0, 2.5 and 3.0 mg/l Fe. Calculate the factor which is
equivalent to mg/l Fe for unit absorbance from the graph.
6. PROCEDURE
Mix sample throughly and measure 50 ml into a 250ml beaker. If the sample contains more
than 3 mg/l iron use a smaller accurately measured portion and dilute to 50ml. Take 50 ml
distilled water as blank. Follow the same procedure used for the standard calibration. Measure
the absorbance against the blank. Note the sample absorbance.
Note If sample is coloured or turbid, carry out a second set of sample through all the steps of
the procedure without adding phenanthroline. Instead of distilled water blank use this to set
photometer to zero absorbance.
7. CALCULATIONS
Total iron, mg/litre Absorbance x F x Dilution (if any) Where,
Reference : APHA 3500 — Fe (D)
GLAB-43 (Revised) - Iron (Phenanthroline method) Page 3 of 3

Page 179
SECTION : WATER
THE DETERMINATION OF IRON (LOW RANGE) (T.P.T.Z. METHOD)

Date issued : 23rdMarch 1995
Page 180

IRON (T.P.T.Z METHOD) (LOW RANGE)
G-LAB METHOD No.44
(By K.P. Abraham)
1. SCOPE
This method covers the determination of iron in all water samples eg. drinking water, cooling
water, waste water, saline water etc. Using this method iron concentrations varying from
0.01 mg/l — 1.0 mg/l can be done using 50 mm cell.
2. PRINCIPLE
The sample is heated in a water bath for one hour with mercapto acetic acid to digest
the insoluble and colloidal iron. Then the solution is buffered to a pH 5.4 and the iron is
complexed with 2,4,6 tripyridyl 1,3,5 triazine solution.This forms a blue coloured
complex which is measured colorimetrically.
3. APPARATUS
3.1. Plastic sample bottles
These should be thoroughly washed and
heated in a solution of 1% mercaptoacetic
acid for 1 hour. This will remove any iron
from the inside of the bottle.
JASCO, Model V 530
3.3. Sample cells, 50 mm
3.4. Pipettes, 1, 2, 4, 5 and 10 ml.
3.5. Volumetric cylinder, 1 litre.
4. REAGENTS
4.1. T.P.T.Z reagent.
Dissolve 0.2 g of 2,4,6 tripyridyl 1,3,5 triazine in 10 ml of 50% hydrochloric acid
(HC1). Make upto 1L.
4.2. Buffer (pH 5.4)

Dilute 253 ml of glacial accetic acid with 300 ml of distilled water. Add 154.5
ml of ammonia solution (sp.gr 0.91) and make up to 1 1 using distilled water.
4.3. Mercapto Acetic Acid Thioglycollic acid.
4.4. Iron solution, standard (1 ml = 1 mg Fe)

Standard iron solution (1 ml = 1 mg Fe) as purchased or dissolve 1.000 g of pure iron
wire in hydrochloric acid and make upto 11
4.5. Iron Solution, Standard (1 ml = 0.01 mg Fe)

Pipette 10.0 ml of the iron standard solution (1 ml = 1 mg Fe) to a 1000 ml volumetric
flask and make up to the mark using distilled water.
GLAB-44 (Revised) - Iron (T.P.T.A method) Page 2 of 3

Page 181
5. CALIBRATION
Pipette 1.0, 2,0, 4.0, 6.0, 8.0 and 10.0 ml of the standard iron solution (1 ml = 0.01 mg Fe) to
clean and specially prepared plastic bottles. Add remaining volume of distilled water to make
the final volume as 100 ml. Take 100 ml distilled water as blank. Follow the same procedure
adopted for the sample described in section 6. Note the absorbances. This will give absorbances
corresponding to 0.1, 0.2, 0.4, 0.6, 0.8 and 1.0 mg/l Fe concentration. Plot a calibration graph
of absorbance against concentration. Calculate the factor F’ from the graph;
Where F’ is the factor corresponding to mg/l Fe producing unit absorbance.
6. PROCEDURE
Take 100 ml of the sample, after shaking the sample thoroughly, to a clean specially prepared
plastic bottle. Add 1 ml of mercaptoacetic acid and close the bottle tightly. Heat it in a water
bath at 80 – 90°C for 1 hour. Then cool and make upto 100 ml, if there is any volume
reduction. Then discard 50 ml of the sample. Add 4.0 ml of the buffer followed by 4.0 ml of
T.P.T.Z reagent. Mix and wait for 10 minutes for colour development. Measure the absorbance
against the blank at 598 nm using 50 mm cells. Take 100 ml distilled water treated the same
way as the sample, as blank.
In G - Station laboratory, by calling method No.7 in JASCO spectrophotometer, all the
analytical parameters will be set in the spectrophotometer and the concentration can be
measured directly.
7. CALCULATION
Iron mg/l Fe = Absorbance x F, where F’ is the factor from the graph.
GLAB-44 (Revised) - Iron (T.P.T.A method) Page 3 of 3

Page 182
SECTION : WATER
THE DETERMINATION OF MOISTURE

Date issued : 25thApril 1995
Page 183

MOISTURE CONTENT AT 105°C
G-LAB METHOD No.45
(By K.P. Abraham)
1. SCOPE
This test method covers the determination of the moisture content in scale samples collected
from the plant. This method also can be used for the determination of moisture content in
process chemicals like soda ash. This determination is used for calculating other analytical
results on a dry basis.
2. PRINCIPLE
The loss in weight of the sample is determined by heating the sample under controlled
conditions of temperature, time and equipment specifications.
3. APPARATUS
3.1. Drying oven with an opening for natural air circulation
and capable of controlling the temperature at 105°C.
3.2. Chemical balance (sensitivity 0.1mg).
3.3. Clean porcelain or platinum dish with cover (50ml

capacity)
4. REAGENTS
None
5. PROCEDURE
Heat the empty dish in the oven for 30 minutes at 105°C. Place the dish in a desiccator and
cover, allowing 15 - 20 minutes for cooling. Weigh the dish, then weigh approximately 5 g of
the representative sample into the dish and place it in the oven without the cover for 4 hours,
again ensuring that the temperature is maintained at 105°C. Replace the cover and cool the dish
in a desiccator. Weigh when the dish is cold.
6. CALCULATIONS
%Moisture = A—B X 100

A
Where,
A = weight of sample taken, and
B = weight of sample after heating
GLAB-45 (Revised) - Moisture content ( At 105°C) Page 2 of 2

Page 184
G -STATION LABORATORY METHOD
SECTION : WATER
THE DETERMINATION OF NITRITE (HIGH RANGE)

Date issued : 21stMarch 1995
Page 185
TEST METHOD FOR COOLING WATER

NITRITES (HIGH RANGE)
G-LAB METHOD No.46
(By K.P. Abraham)
1. SCOPE
This method covers the determination of nitrite present in cooling waters with a nitrite level of
200 — 2500 mg/l nitrite as NO2. Higher concentrations can be done by suitable dilution.
2. PRINCIPLE
Nitrites react in warm acid solution with potassium permanganate solution in accordance with
the following equation:
2 MnO4 + 5N02 + 6H = 2 Mn2 + 5 NO3 + 3H20

The reaction only gives good results in acid solution. But if acid is added to the nitrite solution,
then nitrite is lost as the volatile nitrous acid giving error to the result. For this reason the
sample is titrated to the acidified and warm (40 0C) permanganate solution until the solution is
just decolourised.
3. APPARATUS
3.2. Pipette, 5 ml.
3.5. Hot plate.
3.6. Volumetric flask, 1 litre.
4. REAGENTS
4.1. Potassium permanganete, Standard (0.02M) BDH Convol (Product No.18039) or
equivalent.
Transfer quantitatively and make upto 1 litre with distilled water.
4.2. Sulphuric Acid 0.38M
Take 500 nil distilled water and carefully add 21 ml concentrated sulphuric acid
(H2S04 sp. gr. 1.84) and dilute to 11.
5. PROCEDURE
Pipette 5 ml of potassium permanganate solution (0.02M) to a clean 500 ml conical flask. Add
about 300 ml of Sulphuric acid solution (0.38M) and heat to 40 °C on a hot plate. Take the
sample in a 25 ml burette. Titrate the sample in to the warm KMnO4 Potassium permanganate
solution until the solution is just decolourised. Note the titre reading.
(Note: Towards the end of the titration the reaction in sluggish, so that the nitrite solution must
be added very slowly).
GLAB-46 (Revised) - Nitrite (Titration method) Page 2 of 3

Page 186
6. CALCULATION
0.02M KMnO4 = 0.1N KMnO4
Nitrite as NO2 mg/l = 0.1 x 5 x 23 x 1000 = __11500_______

Titre reading (ml) Titre reading (ml)
Nitrite as NaNO2 mg/l = 0.1 x 5 x 34.5 x 1000 = __ 17250_____

Titre reading (ml) Titre reading (ml)
Reference : Quantitative inorganic analysis by Vogel, Method X,97.
GLAB-46 (Revised) - Nitrite (Titration method) Page 3 of 3

Page 187
G - LABORATORY METHOD
SECTION : WATER
DETERMINATION OR OIL CONTENT

(BY OIL CONTENT ANALYZER)

Date issued : 08thNovember 1995
Prepared by : Abraham & Gengadurai
Page 188

OIL CONTENT (BY OIL CONTENT ANALYSER)
G-LAB METHOD No.47
(By K.P. Abraham & V. Gengadurai)
1. SCOPE
This method is suitable for biological lipids and mineral hydrocarbons. This is also suitable for most
industrial waste waters or treated effluents and saline waters. Oil contamination in sea water can be
detected quickly by this method. A Knowledge of the quantity of oil and grease present is helpful in
proper design and operation of waste water treatment systems, electrolyzers, demineralizing plant etc.
and also may call attention to certain treatment difficulties and operation problems. This method is
not applicable for the measurement of low—boiling fractions that volatalize at temperature below 70
°C.
2. PRINCIPLE
Dissolved or emulsified oil, grease, fats and waxes are extracted from water by intimate contact with
halogenated solvents like trichlorotrifluro ethane (eg: FLON S—316). A nondispersive infrared
(NDIR) analyzer is used to measure the concentration of hydrocarbons in the solvent solution.
3. APPARATUS
3.1. Oil content analyzer by HORIBA Model OCMA 310 —
3.2. Calibration micro syringe, 25 µ1

3.3. Measuring syringe for sample, 20 ml
3.4. Measuring syringe for solvent, 20 ml
3.5. Seperating funnel, 300 ml
3.5. Measuring cylinder, 100 & 200 ml
3.7. Filter paper
3.8. volumetric flasks, 1 L
4. REAGENT
4.1. Extraction solvent FLON S—316
4.2. Hydrochloric acid (141)

Add 25 ml concentrated hydrochloric acid (HC1) to 25 ml demin water and mix.
4.3. Sodium sulphite, Anhydrous (Na2SO4)
4.4. B - Heavy oil as supplied
4.5. Span solution ( 20.0 ppm range)

Take 20.0 p1 of the B—heavy oil in a 1 litre volumetric flask and make upto the mark with the
extraction solvent and shake well. Span adjustment value for this standard is 20.0
4.6. Take 9.0 µl of the B—heavy oil in a 1 litre volumetric flask and make upto the mark with the
extraction solvent and shake well. Span adjustment value for this standard is 4.5
GLAB-47 (Revised) - Oil Content Page 2 of 4

Page 189
5. CALIBRATION
Make the POWER on and allow 60 minutes for warning up. (wait until the warming up indication
(yellow light) goes off. Press [ESC] to lead the screen into measuring mode.
5.1.ZERO CALIBRATION
5.1.1. Turn [EXTRACT COCK] to Close

5.1.2. Turn [DRAIN COCK] to close
5.1.3. Using the measurement syringe (for the solvent), insert 10 ml of pure solvent (S-316)
through the opening above the monitoring window.
5.1.4. Add one drop of Hydrochloric acid (1+1) using the dropper.
5.1.5. Using the measuring syringe (for sample) insert 20 ml oil free water (Cold DM water)
through the opening above the monitor.
5.1.6. Press [EXTRACT] to begin extraction.
5.1.7. After the extraction is complete, wait for the layer separation (30 Sec)
5.1.8. Open the [EXTRACT COCK] to send the solvent layer to the cell.
5.1.9. After 30 second press turn [EXTRACT COCK] to close
5.1.10. Turn [DRAIN COCK] to open.
5.1.11. After the draining is complete, turn [DRAIN COCK] to close
5.1.12. Open the [EXTRACT COCK] to send the solvent layer to the cell a second time.
5.1.13. The measuring value is displayed at the display screen.
5.1.14. After 30 second press [MEASURE] to start the stability judgment. “Measure” blinks
during the stability judgment. When the stability judgment is complete, “HOLD” is lit.
5.1.15. Press [ZERO CAL.] .The calibration data will be displayed and the zero calibration will
start.
5.1.16. Open [DRAIN COCK] to discharge the drains.
Note: Before pressing [MEASURE], make sure that the [EXTRACT COCK] is open
and a 30s time is elapsed after opening the cock.
5.2. SPAN CALIBRATION
5.2.1. Turn [EXTRACT COCK] to Close

5.2.2. Turn [DRAIN COCK] to close
5.2.3. Using the measurement syringe insert 10 ml of Span solution(span liquid prepared with
the same solvent to be used) through the opening above the monitoring window.
5.2.4. Using the measuring syringe (for sample) insert 20 ml oil free water (Cold DM water)
through the opening above the monitor.
5.2.5. Press [EXTRACT] to begin extraction.
5.2.6. After the extraction is complete, wait for the layer separation (30 Sec)
5.2.7. Open the [EXTRACT COCK] to send the solvent layer to the cell.
5.2.8. After 30 second press turn [EXTRACT COCK] to close
5.2.9. Turn [DRAIN COCK] to open.
5.2.10. After the draining is complete, turn [DRAIN COCK] to close
5.2.11. Open the [EXTRACT COCK] to send the solvent layer to the cell a second time.
5.2.12. The measuring value is displayed at the display screen.
5.2.13. After 30 second press [MEASURE] to start the stability judgment. “Measure” blinks
during the stability judgment. When the stability judgment is complete, “HOLD” is lit.
5.2.14. Press [SPAN CAL.] The calibration data will be displayed and the span calibration will
start.
5.2.15. Open [DRAIN COCK] to discharge the drains.

Page 190
Note: Before pressing [MEASURE], make sure that the [EXTRACT COCK] is open
and a 30s time is elapsed after opening the cock.
6. PROCEDURE
6.1. Take 100 ml of sample and add 10 ml of solvent add 2 drops of (1+ 1)HCl and extract it
out side.
6.2. Turn [EXTRACT COCK] to Close
6.3. Turn [DRAIN COCK] to close
6.4. Add the solvent layer and 20 ml extracted sample through the opening above the monitor
window.
6.5. Open the [EXTRACT COCK] to send the solvent layer to the cell.
6.6. After 30 second press turn [EXTRACT COCK] to close
6.7. Turn [DRAIN COCK] to open.
6.8. After the draining is complete, turn [DRAIN COCK] to close
6.9. Open the [EXTRACT COCK] to send the solvent layer to the cell a second time.
6.10. The measuring value is displayed at the display screen.
6.11. After 30 second press [MEASURE] to start the stability judgement. “Measure” blinks during
the stability judgement. When the stability judgement is complete, “HOLD” is lit.
6.12. Note the displayed reading.
6.13. Open [DRAIN COCK] to discharge the drains.
7. CALCULATION
Oil content mg/L = Reading in the instrument x solvent volume / sample volume.
NOTE:
Solvent is harmful to health, avoid skin contact or breathing of the vapours.
Reference: APHA 5520 C

Manual for HDRIBA OCMA - 310

Page 191
G-LABORATORY METHOD
SECTION : WATER
THE DETERMINATION OF pH

Page 192

pH VALUE
G-LAB METHOD No.48
(By K.P. Abraham & V.Gengadurai)
1. SCOPE
This method covers the determination of the pH value of the water sample. Measurement of pH
is one of the most important and frequently used test in water chemistry. At a given temperature
the intensity of the acidic or basic character of a solution is indicated by pH or hydrogen ion
activity.
2. PRINCIPLE
pH measurement is, determination of the activity of the hydrogen ions by potentiometric

measurement. pH is the negative logarithm of the hydrogen ion concentration in terms of the
power of 10.
3. APPARATUS
3.1. pH meter (Micro processor pH meter WTW)
3.2. Temperature sensor WTW Model TFK 150
3.3. pH electrode. Combined pH electrode, WTW (Sentix)
3.4. Volumetric flasks, 100 ml.
4. REAGENTS
4.1. Distilled water with conductivity <2µs/cm.
4.2. Buffer solutions for pH 4.0, 7.0 and 10.0

These buffers to be prepared from the standard pH ampoules or use the ready made
standard buffers.
5. CALIBRATION
Calibration can be carried out using two different sets of buffers. Buffer sets are cd - WTW
standard buffer and ct - WTW technical buffers. Buffer set selection switch is at the back side of
the equipment.
5.1. Select the buffer set to ct.
5.2. Take the first buffer in a beaker and immerse the clean electrode into it and shake slowly.
5.3. Press <CAL> button and bring the display to the Calibration mode.
5.4. Press <RUN ENTER> for the instrument to accept the 1st buffer’s reading.
5.5. When ct2 is requested remove the present buffer clean the electrode and keep the
GLAB-48 (Revised) - pH determination Page 2 of 3
Page 193
second buffer and shake slowly.
5.5. Press < RUN ENTER >. The next display will be the slope. It should have to be
between -62 & -50 mV/pH.
5.7. Press < RUN ENTER>. Now the display will be asymmetric potential and it is
expected to be between + 45 mV.
5.8. Press of < RUN ENTER> and select pH. The instrument is ready for measurement.
6. PROCEDURE
Rinse the electrode with distilled water and then with sample. Take the sample in a beaker,
immerse the electrode and the temperature sensor in it and leave it for few minutes to stabilize.
Note the displayed readings.
7. CALCULATION
Report the meter reading as the pH of the sample at the displayed sample temperature.
8. NOTES:
8.1. pH measurement in low ionic media:
For samples like demineralised water and distillate add 1 ml of neutral KC1 solution to
100 ml of the sample.
8.2. Checking pH electrode efficiency:
8.2.1. Press <M> and bring it to mV.
8.2.2. Keep a 4.00 pH buffer solution at a temperature between 20 and 25°C and
slowly shake the solution. If the mV indication is other than 175+20 mV it
is to be assumed that the electrode is not in the proper order (it may be
polluted or defective). Now go for replacing or cleaning.
8.3. Cleaning of a polluted electrode:
First wash with the demineralised water and then with mild soap solution for few
minutes. Now rinse the electrode with demineralised water several times.
8.4. Storage during measuring pauses:

8.4.1. Do not let the electrode dry.
8.4.2. Immerse in a 3 Molar Potassium Chloride solution.
8.4.3. Never use distilled water for storage.
Reference: APHA 4500 - H+

Manual for WTW .
GLAB-48 (Revised) - pH determination Page 3 of 3

Page 194
SECTION : WATER
THE DETERMINATION OF ORTHO PHOSPHATE (HIGH RANGE)

(ASCORBIC ACID REDUCTION METHOD)

Date issued : 30thOctober 1995
Page 195

ORTHO PHOSPHATE (HIGH RANGE)
(COLORIMETRIC ASCORBIC ACID REDUCTION METHOD)
G-LAB METHOD No.49
(By K.P. Abraham)
1. SCOPE
This method covers the determination ortho phosphates in boiler waters and in waste waters.
The method is usable in the range from 0.50 to 5.0 mg/l of phosphate.
2. PRINCIPLE
Ammonium molybdate and antimony potassium tartrate react with ortho phosphate to form an
antimony-phosphate-molybdate complex. The complex is reduced with ascorbic acid to form a
deep blue coloured molybdenum complex. The colour intensity is proportional to the
phosphorous concentration. Absorbance is measured at a wave length 880 nm.
3. APPARATUS
JASCO model V 530
3.2. Acid-washed glass ware
3.2.1. Ernelmeyer flasks, 250 ml (10 Nos)
3.2.2. Pipettes, 0.5,1, 2, 3, 4 & 5 ml
3.2.3. volumetric flasks, 1 lit, 200 ml, 100 ml (10 Nos.)
3.3. Top pan balance
4. REAGENTS
4.1. Sulphuric acid (~1 M)

Add carefully 56 ml concentrated sulphuric acid (H2S04, Sp.gr 1.84) to 500
ml distilled water and make up to 1 litre.
4.2. Ascorbic acid
Crystalline ascorbic acid (C6H8O6)
4.3. Sodium hydroxide (2M)
Dissolve 80 g of sodium hydroxide (NaOH) in distilled water and make up
to 1 litre.
Dissolve 0.5 g of phenolphthalein in a mixture of 50 ml ethyl alcohol and 50 ml
water.
GLAB-49 (Revised) - Phosphate, Ortho ,High range (Ascorbic acid reduction method) Page 2 of 3
Page 196
4.5. Antimony potassium tartrate molybdate reagent

Dissolve 5.6 g of ammonium molybdate in 700 ml distilled water, add 70 ml
concentrated sulphuric acid (H2S04, sp.gr.1.84) carefully and allow to cool. Add
13g antimony potassium tartrate ([K(Sbo)C4H4O6.½.H20]) to the above solution
and dissolve.Cool to room temperature and make up to 1 litre.
4.6. Phosphate, standard solution (1 ml = 0.05 mg P04)

Dissolve 1.4329 g of potassium dihydrogen phosphate (KH2PO4) that has been
dried for 1 hour in an oven at 1050C, in distilled water and make up to 1 litre. This
will give a stock solution of 1 ml 1 mg P04. Pipette 10.0 ml of this and dilute to
5. CALIBRATION
Pipette 0.5,1.0, 2.0, 3.0, 4.0, and 5.0 mls of the standard phosphate solution
(1 ml = 0.05 mg) into 250 ml Erlenmeyer flask and make up the volume to about 50 ml
with distilled water. Take 50 ml distilled water in one flask as blank. To the above
solution add few drops of phenolphthalein indicator and then add sodium hydroxide
solution (2M) until the phenolphthalein shows pink colouration. Neutralise the excess
alkali with drops of sulphuric acid (1M). Add 10 ml of the antimony potassium tartrate
reagent and then a pinch of ascorbic acid (about 0.2 g). Mix well and make up to 100
ml. Wait for 10 minutes for colour development. Measure the absorbance at 880 nm
using 10 mm cell. Plot a graph with absorbance against phosphate concentration.
Assuming the sample volume as 50 ml, then the standards used in this calibration are
equivalent to 0.5, 1.0, 2.0, 3.0, 4.0 and 5.0 mg/l P04. Calculate the factor mg/l
phosphate/unit absorbance from the graph.
6. PROCEDURE
Take 50 ml of the clear sample in a 250 ml Erlenmeyer flask. If sample contains more than
5mg/l phosphate, use a corresponding smaller sample diluted to 50 ml with distilled water.
Take 50 ml distilled water as blank. To the above solution add few drops of phenolphthalein
indicator and then add sodium hydroxide solution (2M) until the phenolphthalein shows pink
colouration. Neutralise the excess alkali with drops of sulphuric acid (1M). Add 10 ml of the
antimony potassium tartrate reagent and then a pinch of ascorbic acid (about 0.2 g). Mix well
and make up to 100 ml. Wait for 10 minutes for colour development. Measure the absorbance
at 880 nm using 10 mm cell against the blank. Note the absorbance.
Note: For boiler water, the neutralisation step can be omitted and antimony potassium tartrate
reagent can be added followed by ascorbic acid.
In G - Station laboratory, by calling method No.1 in JASCO V 530 spectrophotometer, all the
parameters required for the phosphate determination will be set in the instrument. The
concentration in mg/L can be measured directly. (Note: Follow the JASCO V530
spectrophotmeter routine analytical procedure.)
7. CALCULATION
Phosphate as P04, mg/l Absorbance x F
Where, F is the factor obtained from the calibration graph.

APHA 4500 - P (E)
GLAB-49 (Revised) - Phosphate, Ortho ,High range (Ascorbic acid reduction method) Page 3 of 3
Page 197
SECTION : WATER
THE DETERMINATION OF ORTHO PHOSPHATE (LOW RANGE)

Page 198

ORTHO PHOSPHATE (LOW RANGE)
G-LAB METHOD No.50
(By K.P. Abraham)
1. SCOPE
This method covers the determination ortho phosphates in boiler waters and in waste
waters. The method is usable in the range from 0.01 to 0.5 mg/l of phosphorous.
2. PRINCIPLE
Ammonium molybdate and antimony potassium tartrate react with ortho phosphate to
form an antimony –phosphate-molybdate complex. The complex is reduced with ascorbic
acid to form a deep blue coloured molybdenum complex. The colour intensity is
proportional to the phosphorous concentration. Absorbance is measured at a wave length
880 nm.
3. APPARATUS
JASCO model V 530
3.2.1. Ernelmeyer flasks, 250 ml (10 Nos.)
3.2.2. Pipettes, 0.5,1, 2, 3, 4 & S ml
3.2.3. Volumetric flasks, 1 lit, 100 ml (10 Nos.)
3.3. Top Pan balance
4. REAGENTS
4.1. Sulphuric acid (~1 M)

Add carefully 56 ml concentrated sulphuric acid (H2S04, sp.gr 1.84) to 500 ml
distilled water and make up to 1 litre.
4.2. Ascorbic acid


Dissolve 80 g of sodium hydroxide (NaOH) in distilled water and make up to 1
litre.
GLAB-50 (Revised) - Phosphate, Ortho, Low range (Ascorbic acid reduction method) Page 2 of 3
Page 199

water.
Dissolve 5.6 g of ammonium molybdate in 700 ml distilled water, add 70 ml
concentrated sulphuric acid (H2SO4, sp.gr.1.84) carefully and allow to cool. Add
0.13 g antimony potassium tartrate ([K(Sbo)C4H406.½ H20]) to the above solution
and dissolve. Cool to room temperature and make up to 1 litre.

dried for 1 hour in an oven at 105 0C, in distilled water and make up to 1 litre. This
will give a stock solution of 1 ml = 1 mg P04. Pipette 10.0 ml of this and dilute to
5. CALIBRATION
Pipette 0.5,1.0, 2.0, 3.0, 4.0, and 5.0 mls of the standard phosphate solution (1 ml = 0.01 mg)
into 250 ml Erlenmeyer flask and make up the volume to about SO ml with distilled water.
Take 50 ml distilled water in one flask as blank. To the above solution add few drops of
phenolphthalein indicator and then add sodium hydroxide solution (2M) until the
phenolphthalein shows pink colouration. Neutralise the excess alkali with drops of sulphuric
acid (1M). Add 10 ml of the antimony potassium tartrate reagent and then a pinch of ascorbic
acid (about 0.2 g). Mix well and make up to 100 ml. Wait for 10 minutes for colour
development. Measure the absorbance at 880 nm using 50 mm cell. Plot a graph with
absorbance against phosphate concentration. Assuming the sample volume as SO ml, then the
standards used in this calibration are equivalent to 0.1, 0.2, 0.4, 0.6, 0.8 and 1 mg/l P04.
Calculate the factor mg/l phosphate/unit absorbance from the graph.
6. PROCEDURE
Take 50 ml of the clear sample in a 250 ml Erlenmeyer flask. If sample contains more than 1
mg/l phosphate, use a corresponding smaller sample diluted to 50 ml with distilled water. Take
50 ml distilled water as blank. To the above solution add few drops of phenolphthalein
indicator and then add sodium hydroxide solution (2M) until the phenolphthalein shows pink
colouration. Neutralise the excess alkali with drops of sulphuric acid (lM). Add 10 ml of the
antimony potassium tartrate reagent and then a pinch of ascorbic acid (about 0.2 g). Mix well
and make up to 100 ml. Wait for 10 minutes for colour development. Measure the absorbance
at 880 nm using 50 mm cell against the blank. Note the absorbance.
In G-Station laboratory, by calling method No.8 in JASCO spectrophotometer, all the

concentration in mg/L can be measured directly. (Note: Follow the JASCO spectrophotometer
routine analytical procedure.)
7. CALCULATION
Phosphate as P04, mg/l = Absorbance x F
Reference ASTM D 515

APHA 4500-P (E)
GLAB-50 (Revised) - Phosphate, Ortho, Low range (Ascorbic acid reduction method) Page 3 of 3
Page 200
SECTION : WATER
THE DETERMINATION OF ORTHOPHOSPHATE (HIGH RANGE)
(MOLYBDOVANADO PHOSPHATE METHOD)

Date issued : 22ndMarch 1995
Page 201

ORTHOPHOSPHATE ( MOLYBDOVANADO PHOSPHATE METHOD)
G-LAB METHOD No.51
(By K.P. Abraham)
1. SCOPE
This method covers the routine determination of dissolved orthophosphate in water. It is

best suited to boiler water and boiler feed water containing 1 to 100 mg/l of
orthophosphate. The range can be extended upward by diluting the sample.
2. PRINCIPLE
In a dilute orthophosphate solution, ammonium molybdate reacts under acid conditions to

form a heteropoly acid, molybdophosphoric acid. In the presence of vanadiam, yellow
vanadomolybdo phosphoric acid is formed. The intensity of the yellow colour is
proportional to phosphate concentration. Its absorbance is measured at a wave length of
425 nm.
3. APPARATUS
JASCO, Model V 530

All glassware and sampling bottles shall be soaked in hydrochloric acid (1+3) to
remove possible phosphate coating, and rinsed with distilled water prior to use.
3.2.1. Volumetric flasks, 50 ml, 10 Nos.
3.2.2. Volumetric flask, 1 litre.
3.2.3. Pipettes, 0.5, 1, 2, 5 and 10 ml.
3.3. Plastic cups, 10 Nos.
3.4. Cells, 10 mm.
4. REAGENTS
4.1. Ammonium molybdate — Vanadate solution

Dissolve 25 g of ammonium molybdate [(NH4)6MO7O24.4H2O] in 400 ml of
distilled water. Dissolve 1.25 g of ammonium metavanadate (NH4VO3) in a
mixture of 300 ml of water and 100 ml of concentrated sulphuric acid (sp. gr.
1.84). Add the first solution to the second solution, mix and dilute to 1 litre with
distilled water.
4.2. Phosphate solution standard (1 ml = 1 mg P043-)

dried for 1 hour in an oven at l05C, in distilled water and make up to exactly 1
litre in a volumetric flask.
GLAB-51 (Revised) - Phosphate, Ortho, High range (Vanadate method) Page 2 of 3

Page 202
5. CALIBRATION
Prepare a series of standard phosphate solutions to cover the range from 10 to 100 mg/l of
phosphate. Prepare the standards by diluting suitable volumes of the standard phosphate
solution (1 ml = 1 mg P04) to SO ml with distilled water. One miili litre of the phosphate
standard solution diluted to 50 ml produces a standard containing 20 mg/l of phosphate.
Pipette 0.5, 1, 2, 3, 4 and 5 ml of the standard phosphate solution to 50 ml volumetric
flasks and make upto the mark with distilled water. This will provide standards of 10, 20,
40, 60, 80 and 100 ppm phosphate. Transfer these to 6 plastic cups and take 50 ml
distilled water in another cup.
Add 10 ml ammonium molybdatevanadate reagent to each. Wait for 30 minutes and

measure the absorbance at 425 nm using 10 mm cells. Plot the absorbance values
obtained as ordinates and the corresponding phosphate concentration as abscissas.
Calculate a factor ‘F’ from the graph, which is equal to mg/l phosphate per unit
absorbance.
6. PROCEDURE
Take 50 ml of clear sample (filter if suspended matter is present) in a clean beaker. If

sample contains more than 100 mg/l phosphate, use a correspondingly smaller sample
diluted to 50 ml with distilled water. Take SO ml distilled water as blank. Add 10 ml each
ammonium molybdate vanadate reagent to the sample and blank. Wait for 30 minutes
for colour development. Measure the absorbance of the sample against blank at 425 nm
wave length using 10 mm cell. Record the absorbance.
7. CALCULATION
Orthophosphate, mg/l (ppm) = Absorbance x F
Where,
‘F’ is the factor calculated from the calibration graph.
Reference APHA 4500 - P (C)
GLAB-51 (Revised) - Phosphate, Ortho, High range (Vanadate method) Page 3 of 3

Page 203
SECTION : WATER
THE DETERMINATION OR ORTHO PHOSPHATES
(STANNOUS CHLORIDE REDUCTION METHOD)

Page 204

ORTHOPHOSPHATE (STANNOUS CHLORIDE REDUCTION METHOD)
G-LAB METHOD No.52
(By K.P. Abraham)
1. SCOPE
This colourimetric method covers the determination of orthophosphates in water and waste
water. Phosphorous is widely distributed in the environment both in water and waste water as
inorganically and organically bound phosphates. Phosphates are widely used as boiler water
conditioners and testing of phosphate is an essential step in the control of boiler water quality.
2. PRINCIPLE
Phosphates present in water are in two forms:—

1. Ortho phosphates or reactive phosphates
2. Hydrolyzable phosphates ie. meta, pyro, and tn polyphosphates.
The second form of phosphates has to be converted to orthophosphates to enable it to be
tested using the orthophosphate testing procedures. On addition of ammonium molybdate,
orthophosphates and molybdate ions condense in acidic solutions to give molybdophosphoric
acid (phosphomolybdic acid) which upon reduction with stannous chloride is turned to
intensely coloured molybdenum blue of uncertain composition. The intensity of the blue
colour is proportional to the amount of phosphate initially incorporated to the heteropoly acid
in the solution.
Interferences:
Silica and arsenate give positive interference only if the sample is heated. Negative
interferences are caused by arsenate, fluoride, thorium, bismuth, sulfide, thiosulfate, ferrous
iron >100 mg/L, thiocyanate or excess molybdate. Sulphide interference may be removed by
bromine water. Many other ions do not interfere if the concentration do not exceed 1000
mg/L. If HNO3 is used in the test, chloride interferes above 75 mg/L.
3. APPARATUS
3.1. Spectrophotometer JASCO model V 530 or Filter

photometer WTW model MPH 1500 with 590 filter.
3.2. 10 mm cuvette cell.
3.3. Pipettes, conical flasks, volumetric flasks etc.
4. REAGENTS
4.1. Ammonium molybdate solution (1.5%)
Dissolve 15 grams of ammonium molybdate [(NH4)2MO7O24.4H20]
in 150 ml of demin water, add this to a solution containing 182 ml sulphuric acid
and 600 ml demin water and make up to 1 litre.
4.2 Stannous chloride solution (2%).
Dissolve 1 gram of stannous chloride (SnCl2.2H2O) in 5 ml of concentrated HC1
and dilute to 50 ml with demin water. Add a few grains of tin metal into the solution
and store in a fridge at 4° C.
4.3 Strong acid solution (H2SO4 + HNO3)
Slowly add 30 ml concentrated sulphuric acid to about 60 ml demin water. When
cool and 0.4 ml of concentrated nitric acid and dilute to 100 ml.
GLAB-52 (Revised) - Phosphate, Ortho (Stannous chloride method) Page 2 of 3

Page 205
4.4 Phosphate standard stock solution 1000 mg/L as PO43-

Dissolve 1.4329 grams of KH2PO4 (which has been dried at 105 °C for 1 hour in an
oven) in demin water and make up to 1 litre or take standard 1000 mg/L (ready
made) phosphate solution (lml= 1 mg of P043-)
4.5 Phosphate standard solution (lml 0.100 mg P043-)
Pipette 10 ml from the stock solution and make up to 100 ml.
5. CALIBRATION
Follow the procedures given in section 6 (and the specific procedures of the instruments used)
using the standards prepared as follows. Pipette 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5 and 5.0
ml of standard phosphate solution and make up to 50 ml. The above solutions are
corresponding to 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10.0 mg/L of PO43- respectively.
Plot a graph using concentration versus absorbance and calculate the factor F for unit
absorbance.
6. PROCEDURE
6.1 Take 50 ml of the sample and neutralyze with strong acid solution (only if the
sample is highly alkaline) using phenolphthalein indicator. If more than 5 drops acid
is required take a smaller sample and dilute to 50 ml with demin water.
6.2 Add 5 ml of ammonium molybdate solution and mix well.
6.3 Add 0.25 ml of stannous chloride solution and mix well.
6.4 Wait for 10 minutes and measure the absorbance at 700 nm using 10 mm cell.
6.5 Blank is carried out using the same procedure but using 50 ml of demin water.
Note: (Follow the specific instruments’ operation procedures for the measurements
and the determination of the concentrations).
7. CALCULATION
If JASCO spectrophotometer is used read the concentration mg/L

as concentration of PO43-
If filter photometer MPH 1500 is used Concentration of PO43-, mg/L

Absorbance X F
References: APHA 4500 - P D
GLAB-52 (Revised) - Phosphate, Ortho (Stannous chloride method) Page 3 of 3

Page 206
SECTION : WATER
THE DETERMINATION OF ORTHO & META PHOSPHATE (LOW RANGE)


Page 207

ORTHO AND META PHOSPHATE (LOW RANGE)
G-LAB METHOD No.53
(By K.P. Abraham)
1. SCOPE
This method covers the determination of specified forms of phosphorous compounds in waters
and in waste waters. This method can be used for the determination of different inhibitors,
which contains polyphosphates. The method is usable in the range from 0.01 to 0.5 mg/l of
phosphorous.
2. PRINCIPLE
Ammonium molybdate and antimony potassium tartrate react with ortho phosphate to form an
antimony-phosphate-molybdate complex. The complex is reduced with ascorbic acid to form a
deep blue coloured molybdenum complex. The colour intensity is proportional to the
phosphorous concentration. Phosphates other than orthophosphates are hydrolysed using
potassium persulphate and sulphuric acid before the colour development. Absorbance is
measured at a wave length 880 nm.
3. APPARATUS
3.1. Spectrophoto meter
JASCO Model V 530
3.2. Acid- washed glass ware
3.2.1. Erlenmeyer flasks, 250 ml (10 Nos.)
3.2.2. Pipettes, 1, 2, 4, 5 & 10 ml
3.2.3. Volumetric flasks, 1 lit, 100 ml (10 Nos.)
3.3. Hot plate
4. REAGENTS
4.1. Potassium persulphate
Crystalline potassium persuiphate (K2S2O5)
4.2. Sulphuric acid (1 M)

Add carefully 56 ml concentrated sulphuric acid (H2SO4, sp.gr 1.84) to 500 ml distilled
water and make up to 1 litre.
4.3. Ascorbic acid


Dissolve 80 g of sodium hydroxide (NaOH) in distilled water and make up to 1 litre.

Dissolve 0.5 g of phenolphthalein in a mixture of 50 ml ethyl alcohol and 50 ml water.
GLAB-53 (Revised) - Phosphate, Ortho+Meta, Low range (Ascorbic acid reduction method) Page 2 of 3
Page 208

Dissolve 5.6 g of ammonium molybdate in 700 ml distilled water, add 70 ml concentrated
sulphuric acid (H2SO4, sp.gr.1.84) carefully and allow to cool. Add 0.13 g antimony
potassium tartrate ([K(Sbo)C4H406 ½ H20]) to the above solution and dissolve.Cool to
room temperature and make up to 1 litre.

Dissolve 1.4329 g of potassium dihydrogen phosphate (KH2PO4) that has been dried for 1
hour in an oven at 105°C, in distilled water and make up to 1 litre. This will give a stock
solution of 1 ml = 1 mg P04. Pipette 10.0 ml of this and dilute to 1000 ml in a volumetric
flask.
5. CALIBRATION
Pipette 1.0, 2.0, 4.0, 6.0, 8.0 and 10.0 mls of the standard phosphate solution (1 ml = 0.01 mg)
into 250 ml Erlenmeyer flask and make up the volume to about 100 ml with distilled water.
Take 100 ml distilled water in one flask as blank. Add about 1 g each of potassium persulphate
and 4 ml of H2S04 (1N). Boil on hot plate for 30 minutes and cool. To the above solution add
few drops of phenolphthalein indicator and then add sodium hydroxide solution (2M) until the
acid (1N). Add 10 ml of the antimony potassium tartrate reagent and then a pinch of ascorbic
development. Measure the absorbance at 880 nm using 50 mm cell. Plot a graph with
absorbance against phosphate concentration. Assuming the sample volume as 100 ml, then the
standards used in this calibration are equivalent to 0.1, 0.2, 0.4, 0.6, 0.8 and 1 mg/l P04.
Calculate the factor mg/l phosphate/unit absorbance from the graph.
6. PROCEDURE
Take 100 ml of the clear sample in a 250 ml Erlenmeyer flask. If sample contains more than 1
mg/l phosphate, use a corresponding smaller sample diluted to 100 ml with distilled water.
Take 100 ml distilled water as blank. Add about 1 g each of potassium persuiphate and 4 ml of
H2S04 (1M). Boil on hot plate for 30 minutes and cool. To the above solution add few drops of
phenolphthalein indicator and then add sodium hydroxide solution (2M) until the
acid (1N). Add 10 ml of the antimony potassium tartrate reagent and then a pinch of ascorbic
development. Measure the absorbance at 880 nm using 50 mm cell. Note the absorbance,
In G-Station laboratory, by calling method No.1 in JASCO spectrophotometer, all the

concentration in mg/L can be measured directly.
(Note: Follow the JASO spectrophotmeter routine analytical procedure.)
7. CALCULATION
Phosphate, mg/l = Absorbance x F


APHA 4500 - P (E)
GLAB-53 (Revised) - Phosphate, Ortho+Meta, Low range (Ascorbic acid reduction method) Page 3 of 3
Page 209
SECTION : WATER
THE DETERMINATION OF PHOSPHONATES

Date issued : 21stMarch 1995
Page 210

PHOSPHONATES IN COOLING WATER
G-LAB METHOD No.54
(By K.P. Abraham)
1. SCOPE
This test method has been developed for the determination of phosphonates and there by to
provide control over phosphonates used in scale and corrosion inhibitors.
2. PRINCIPLE
The phosphonate is oxidised with potassium persulphate and sulphuric acid and the
resulting orthophosphate is determined by the vanadomolydate method using a suitable
spectrophotometer. In vanadomolydate method orthophosphate reacts with ammonium
molybdate to form molyhdophosphoric acid. In the presence of vanadium, yellow
vanadomolybdophosphate acid is formed and its absorbance is measured at the wavelength
420 nm.
3. APPARATUS
JASCO, Model V 530
3.2. Hot plate
3.3. Funnel
3.4. Filter paper (No. 41)
3.5. Top pan weighing balance
3.6. Conical flasks, 250 ml, 10 Nos.
3.7. Pipettes, 10, 20 and 50 ml.
3.8. Volumetric flasks, 100 ml, 10 Nos.
3.9. Volumetric flask, 1 litre.
3.10. Cells, 10 mm.
4. REAGENTS
4.1. Potassium persulphate.

Crystalline potassium persulphate (K2S208).
4.2. Sulphuric Acid solution (2N)

Cautiously add 56 ml of concentrated sulphuric acid (H2S04 Sp.gr.1.84) to about
500 ml distilled water with stirring. Cool and dilute to 1 litre.
4.3. Vanadomolybdate Reagent

Dissolve 20 g ammonium molybdate tetrahydrate in about 250ml distilled water.
Dissolve 1.000 g ammonium metavanadate in about 200 ml distilled water
acidified with 40 ml concentrated nitric acid (HN03 Sp.gr.1.42). Mix the two
solutions add 100 ml concentrated nitric acid and dilute to 1 litre with distilled
water. Store in polythene bottle.
GLAB-54 (Revised) - Phosphonate (P2O5) Page 2 of 3

Page 211
4.4. Standard phosphonate solution (lml=lmg P205)

Dissolve 1.9175 g monopotassium dihydrogen phosphonate (Previously dried for
1 hour at 105 0C) in distilled water. Transfer to a 1 litre volumetric flask and
dilute to the mark with distilled water.
4.5. Standard phosphonate solution (lml = l0µg P205)

Pipette 10.0 ml standard phosphonate solution (lml = 1mg P205) into 1 litre
volumetric flask and dilute to the mark with distilled water.
5. CALIBRATION
Into 5 clean 100 ml volumetric flasks, pipette 10.0, 20.0, 30.0, 40.0 and 50.0 ml of the
standard phosphonate solution (lml = 10 µg P205). Pipette into these and into an empty 100
ml flask 25.0 ml of vanadomolybdate reagent. Dilute each to the mark with distilled water
and mix well. Measure the absorbance at 420 nm using 10 mm cell against the prepared
reagent blank. Plot a calibration graph of absorbance versus concentration of P205 mg/I.
Assuming the sample volume as 50 ml, then the quantities used in this calibration are
equivalent to 2,4,6,8 and 10 ppm. Calculate the factor ‘F’ which is mg/l P205 equivalent to
get unit absorbance.
6. PROCEDURE
Pipette 50 ml sample and 50 ml distilled water as blank to two 250 ml conical flasks. Add
to each 5m1 1M sulphuric acid and 1 g potassium persulphate crystals. Boil gently for 15 -
20 minutes. Cool and filter through a filter paper (No.41) if any suspended particles
remains. Transfer quantitatively to a 100 ml volumetric flask, add 25 ml vanadomolybdate
reagent, make up to the mark with distilled water and mix well. Measure the absorbance of
the sample against the blank at 420 mm wavelength using 10 mm cells. Note the
absorbance.
7. CALCUATION
Phosphonate as P205, mg/l = Absorbance x F, where F is the factor from the

calibration graph.
Reference : Aquarite Cooling Water treatment by Albright & Wilson.
GLAB-54 (Revised) - Phosphonate (P2O5) Page 3 of 3

Page 212
SECTION : WATER
THE DETERMINATION OF POLYPHOSPHATE BASED INHIBITORS

Page 213

POLYPHOSPHONATE BASED INHIBITORS IN RECIRCULATED BRINE
G-LAB METHOD No.55
(By K.P. Abraham)
1. SCOPE
This test method covers the determination of polyphosphate based inhibitors in sea water,
recirculated brine or in the blow down of evaporators. This is a modified method to suit the
determination of specified inhibitors like Albrivap DSB, Calgon EL2438 etc.
2. PRINCIPLE
Polyphosphonate based inhibitors contains polythosphonates which can be hydrolysed to bring
it down to orthophosphates. The orthophosphates formed will be proportional to the inhibitor
concentration. Ammonium molybdate and antimony potassium tartrate react with
orthophosphate to form an antimony phosphate molybdate complex. The complex is reduced
with ascorbic acid to form a deep blue coloured molybdenum complex. The colour intensity is
proportional to the phosphate concentration and there by proportional to the inhibitor
concentration. Absorbance is measured at a wave length 880 nm.
3. APPARATUS
JASCO, Model V 530
3.2. Acid-washed glass ware
3.2.1. Erlenmeyers flasks, 250 ml (10 Nos.)

3.2.2. Volmetric flasks, 100 ml (10 Nos), 1 litre.
3.2.3. Measuring cylinder, 100 ml.
3.2.4. Spectrophotometer cells, 10 mm.
3.3. Hot plate
3.5. Weighing boats
4. REAGENTS
4.1. Potassium persulphate

Crystalline potassium persulphate (K2S208)
4.2. Sulphuric acid (2N)

Add carefully 56 ml concentrated sulphuric acid (H2S04, sp.gr 1.84) to 500 ml
distilled water and make up to 1 litre.
4.3. Ascorbic acid.

Crystalline ascorbic acid (C6H8O6).
GLAB-55 (Revised) - Polyphosphate base inhibitors Page 2 of 3

Page 214
4.4. Sodium hydroxide (2N)

Dissolve 80 g of sodium hydroxide (NaOH) in distilled water and make up to 1 litre.

water.
4.6. Antimony potassium tartrate molybdate reagent.

Dissolve 5.6 g of ammonium molybdate in 700 ml distilled water. Add 70 ml
concentrated sulphuric acid (H2S04, sp.gr.1.84) to this carefully an al1ow to cool. Add
0.13 g antimony potassium tartrate ([K(SbO)C4H406½H2]) to the above solution
dissolve. Cool to room temperature and make up to 1 litre.
4.7. Inhibitor standard (lml = 1mg inhibitor)

Weigh 1.000 g of the inhibitor, which is to be determined in the sample (Eg. Albrivap
DSB, Calgon EL 2438 etc). Dilute this with filtered sea water and make upto 1 litre in
a 1 litre flask. This will give a stock solution of lml = 1mg inhibitor.
4.8. Working standard solution (lml=0.lmg inhibitor)

Pipette 10.0 ml of the stock solution (lml= lmg inhibitor) to 100 ml volumetric flask
and make up to the mark using filtered sea water.
5. CALIBRATION
Pipette 1.0, 2.0, 4.0, 6.0 8.0, and 10.0 ml of the standard inhibitor solution (lml = 0.1mg) into
250 ml Erlenmeyer flask and make up the volume to about 100 ml with filtered sea water. Take
100 ml filtered sea water as blank. Add to each 1 g potassium persulphate and then 4ml
H2S04(2N). Boil on hot plate till the volume reduces to about 50m1. Cool the solution and then
add few drops of phenolphthalein indicator. Add sodium hydroxide solution (2N) until the
phenolphthalein shows pink colouration. Neutralize the excess alkali with drops of sulphuric
acid (2N). Add 10 ml antimony potassium tartrate reagent and then add a pinch of ascorbic acid
(about 0.2 gm). Mix well and make upto 100 ml. Wait for 10 minutes for colour development.
Measure the absorbance at
880 mm wavelength using 10 mm cell against the sea water blank. Plot a graph with
absorbance against the inhibitor concentration in mg/l. of the inhibitor for unit absorbance.
6. PROCEDURE
Take 100 ml of the clear sample in a 250 ml Erlenmeger flask. Take 100 ml clear sea water as
blank. Follow the same procedure used for the calibration. Measure the sample absorbance
against the sea water blank. Note the absorbance.
7. CALCULATION
Inhibitor, mg/l = Absorbance x F
Where,
Absorbance = Sample absorbance against the sea water blank.
Reference : ASTM D 515 , APHA 4500 - P (E)
GLAB-55 (Revised) - Polyphosphate base inhibitors Page 3 of 3

Page 215
SECTION : WATER
QUALITY CHECKS, ACIDITY & ALKALINITY BY AUTOTITRATOR

Date issued : 30thJanuary 1996
Page 216

QUALITY CHECK, ACIDITY & ALKALINITY BY AUTOTITRATOR
G-LAB METHOD No.56
(By K.P. Abraham)
1. SCOPE
This method covers the determination of the purity of different chemicals like, commercial
hydrochloric acid, caustic, ammonia, trisodium phosphate, disodium phosphate, hydrazine etc.
This method also can be used for the concentration determination of different chemicals, p-
alkalinity and rn-alkalinity of water, and for the determination of total acid number and total
base number of oils.
2. PRINCIPLE
The titrations are done automatically using titrant stored in a storage bottle, to the sample taken
in the titration vessel using an interchangeable burette operated by piston and stopcock. The
end point determination is done by the use of specific electrodes like combination universal
glass electrode for acid-base titrations, combination platinum ring electrode for redox titrations,
combination silver ring electrode for argentometric and complexometric titrations, combination
glass electrode for acid-base titrations in non-aqueous media etc. The dispensing and endpoint
determinations are controlled by the analytical conditions stored in the method, which is being
used.
The METTLER DL 25 titrator is a complex analysis station for titrimetric analysis. This titrator
enable the titration to be performed and evaluated automatically. Considerations of the sample
weight in the result calculation allows the analysis to be reported in the desired unit. The setup
procedure for a particular analysis avoids the use of technical instrument parameters. The
titration can be carried out with optimum accuracy using just the standard configuration.
3. APPARATUS
3.1. Auto titrator METTLER DL 25

3.2. Electrodes
3.2.1 Combination universal glass electrode DG111- SC
3.2.2 Combination universal glass electrode
with sleeve diaphram DG115 - SC
3.2.3 Combination platinum ring electrode DG14O - SC
3.2.4 Combination silver ring electrode DG141 - SC
3.3. Titration vessels
3.3.1 Disposable titration beakers,
Polypropylene, 100 ml
3.3.2 Titration vessel, glass, 250 ml
3.4. Pipettes, 2, 5, & 10 ml
3.7. Wash bottle
GLAB-56 (Revised) - Quality checks, Acidity & Alkalinity (Autotitrator method) Page 2 of 5
Page 217
4. REAGENTS
4.1. Sulphuric acid, standard (1.0 N)

water.
4.2. Sodium hydroxide, standard (1 N)

Dissolve 40.0 g of sodium hydroxide (NaOH pellets) in 400 ml distilled water cool
and dilute to 1 L. Standardize using the standard 1.0 N sulphuric acid solution.
4.3. Standard acid solution (0.02 N H2SO4)
a 500 ml volumetric flask and make upto the mark with distilled water.
4.4. Titration solvent
Add 500 ml of toluene and 5 ml of water to 495 ml of anhydrous isopropyl alcohol.
The titration solvent should be made up in large quantities, and its blank value
determined by titration prior to use.
4.5. Alcoholic KOH, 0.1 N
Add 6 g of potassium hydroxide (KOH) to approximately 1 L of anhydrous isopropyl
alcohol. Boil gently for 10 mm to effect solution. Allow the solution to stand for 2
days and then filter the supernatant liquid through a fine sintered glass funnel. Store
the chemical in a chemically resistant bottle. Standardize frequently enough to detect
normality changes of 0.0005 by potentiometric titration of weighed quantities of
potassium acid phthalate in C02- free water.
4.6. Alcoholic KOH, 0.01 N
Take 100 ml of the alcoholic KOH 0.1N and dilute to 1 L using anhydrous isopropyl
alcohol. Standardize this solution prior to use.
4.7. Iodine, standard 0.1 N
To be prepared from an ampoule.
4.8. Sodium bicarbonate
Sodium bicarbonate (NaHCO3) crystals as purchased.
4.9. Buffer solutions (4, 7, & 10)
Ready made buffer solutions or solutions prepared from ampoules to be used.
4.10. 2,4,6 Trimethyl pyridine (Collidine), (CH3)3C5H2N As purchased
4.11. m - Nitrophenol, NO2C6H4OH As purchased.
5. CALIBRATION
Follow the calibration procedures for the pH electrodes as described in the instrument
calibration procedure.
Standardize the solutions used for the analysis as described in the instrument concentration
determination procedures.
5.1. Concentration determination
5.1.1. Switch ON the instrument and the printer.
5.1.2. Enter the DATE, MONTH, and YEAR and confirm each by pressing
RUN.
5.1.3. Enter the method number from the table and press METHOD.
5.1.4. Replace the storage bottle with the titrant for the particular method.
5.1.5. Connect the titration vessel in the titration head.
5.1.6. Enter 15/30 and press BURET mL. This will complete the rinsing of the
burette with the titrant three times.
5.1.7. Replace the titration vessel with wash beaker filled upto 100 ml with DM
water/solvent.
5.1.8. Enter 3 and then press pH Calib., and after 10 seconds press RESET.
Page 218
5.1.9. Pipette the specified volume of the standard solution or specified weight of
the solid standard to a clean 20 ml titration vessel. Add about 10 ml of CM
water if solid sample is taken. Fix the titration vessel in the titration head.
5.1.10. Press MODE and press RUN.
5.1.11. Enter the volume/weight of the standard taken and press RUN.
5.1.12. Press RUN twice again.
5.1.13. Wait till the titration is complete, which is signalled by an audio sound.
The result is displayed and printed.
5.1.14. Enter the method number and press SAVE twice to save the value along
with the method.
5.2. Blank determination
5.2.1. Switch ON the instrument and the printer.
5.2.2. Enter the DATE, MONTH, and YEAR and confirm each by pressing
RUN.
5.2.3. Enter the method number from the table and press METHOD.
5.2.4. Replace the storage bottle with the titrant for the particular method.
5.2.5. Connect the titration vessel in the titration head.
5.2.6. Enter 15/30 and press BURET mL. This will complete the rinsing of the
burette with the titrant three times.
5.2.7. Replace the titration vessel with wash beaker filled upto 100 ml with DM
water/solvent.
5.2.8. Enter 3 and then press pH Calib., and after 10 seconds press RESET.
5.2.9. Take the specified volume of the blank water/solvent in a clean titration
vessel and fix it in the titration head.
5.2.10. Press MODE twice.
5.2.11. Press RUN three times.
5.2.12. Wait till the titration ends.
5.2.13. When the blank determination is over, it is signalled by an audio signal and
the result will be shown on the display and it will be printed. The blank
value will be automatically entered in the program.
5.2.14. Enter the method number and press SAVE twice to save this value along
with the method.
6. PROCEDURE
6.1. Switch ON the instrument and the printer.

6.2. Enter the DATE, MONTH, and YEAR and confirm each by pressing RUN.
6.3. Enter the method number from the table and press METHOD.
6.4. Replace the storage bottle with the titrant for the particular method.
6.5. Connect the titration vessel in the titration head.
6.6. Enter 15/30 and press BURET mL. This will complete the rinsing of the burette with
the titrant three times.
6.7. Replace the titration vessel with wash beaker filled upto 100 ml with DM
water/solvent.
8.8. Enter 3 and then press pH Calib., and after 10 seconds press RESET.
6.9. Clean the titration vessel and take the recommended weight/volume of the sample, as
specified in the table, in the titration vessel.
6.10. Add sufficient DM water/solvent to the titration vessel so that the electrode will
remain immersed properly in the solution.
6.11. Replace the wash beaker with the titration vessel.
6.12. Press RUN.
6.13. Enter the weight or volume and press RUN.
Note: For method No.1, enter the weight as 1.
6.14. Enter the sample identity number as follows and press RUN.
0000 — 0999 — Non routine samples
*000 — *999 — All other methods (* Method No.)
Page 219
6.16. When the BUSY light is blinking press RUN.

6.16. Wait for the titration to finish.
6.17. The resultwill be displayed as well as printed. Additional information and other
titration details can be printed by using the code operations.
6.18. After completing the analysis, replace the titrant with DM water/solvent.
6.19. Carry out the rinsing as in step No.6.6.
6.20. Replace the titration vessel with the wash beaker filled with DM water.
6.21. Switch OFF the instrument.
TABLE
Method Approx. Sample

Analysis for Titrant used Result as Remarks
No. Samplewt/Vol Identity
Alkalinity 1 H2SO4,0.02N 200ml 1000-1999 Mg/l as CaCo3 P&M.Alk
D.S.P 2 HCl, 1.0N 1.0 gram 2000-2999 %w/w Na2HPo4 x 0.6690PO4

x 0.4329 P2O5
T.S.P 3 HCl, 1.0N 0.5 gram 3000-3999 %w/w Na3Po4
x 0.5793PO4
HCl 4 NaOH, 1.0N 1.0ml 4000-4999 %w/v as HCl /SG = %w/w
NaOH 5 HCl, 1.0N 0.5 gram 5000-5999 %w/w NaOH x SG = %w/v
NH3 6 HCl, 1.0N 1.0ml 6000-6999 %w/v as NH3 x SG = %w/w
N2H4 7 I2, 0.1N 25ml 7000-7999 %w/w as N2H4 2g to 2L(Dil)
Na3PO4+NaOH 8 HCl, 1.0 N 20 ml 8000-8999 mmol/L Note(* ,**)
TAN 11 Alk.KOH,0.1N 20 gram 11000-11999 mgKOH/g Tot.Acid.No
Note : (*) NaOH as % w/v = (R high pH mmol/L — R low pH mmol/L) x 0.004

(**) Na3PO4 as % w/v = R low pH mmol/L x 0.0164
7. CALCULATIONS
The calculated result in the specified unit along with the analytical parameters are printed out
as a report after completing the analysis by the titrator. Additional informations, titration curve,
first derivative of the titration curve, table of measured values of the titration curve also can be
printed out from the stored data.
Reference: ASTM D664

APHA 2320
Vogel’s textbook of quantitative inorganic analysis
Manual for METTLER DL 25 Titrator
Page 220
SECTION : WATER
QUALITY CHECK OF LIME STONE

Date issued : 24thFebruary 1996
Page 221

QUALITY CHECK OF LIME STONE
G-LAB METHOD No.57
(By K.P. Abraham)
1. SCOPE
This method covers the determination of calcium content in the lime stone sample and thereby
determine the purity of the limestone. This quality determination is of much importance for the
assessment of the blending plant and for the quality assurance of the chemical and drinking
water. High level of impurities will lead to accumulation of sludge in the lime stone filters.
2. PRINCIPLE
The sample is crushed and brought to solution by hydrochloric acid. Then the calcium
is determined by titration with standard EDTA solution at a pH of 12 -13 in the
presence of Nana indicator. At the end point the indicator changes its colour from
reddish purple to blue.
3. APPARATUS
3.3. Measuring cylinders, 100 ml and SD ml
3.4. Volumetric flasks, 1 litre and 500 ml & 250 ml
3.5. Beaker, 250 ml
3.6. Pipette, 10 ml
3.8 Electric grinder
3.9. Drying oven
3.10. Hot plate
4. REAGENTS
4.1. Disodium ethylenediamine tetra acetate (Na2H2EDTA) solution Standard (0.01M).

Weigh 3.723 g analytical reagent — grade disodium ethylenediaminetetra
acetate dihydrate (EDTA) and dissolve in distilled water and dilute to
1000 ml. Standardize against standard calcium or zinc chloride solution.
4.2. Standard calcium solution (1 ml 1 mg CaCO3)
Weigh 1.000 g anhydrous CaCO3 powder (primary standard or special reagent low in
heavy metals, alkalies and magnesium) into a 500 ml erlenmeyer flask. Place a funnel
in the flask neck and add, a little at a time, 1+1 HC1 until all CaCO3 has dissolved.
Add 200 ml distilled water and boil for a few minutes to expel CO2. Cool, add a few
GLAB-57 (Revised) - Quality check of Lime stone Page 2 of 3
Page 222
drops of methyl red indicator, and adjust to the intermediate orange colour by adding
3N NH4OH or 1+1 HC1, as required. Transfer quantitatively and dilute to 1 litre with
distilled water.
4.3. Calcium hardness buffer (pH 12 - 13)
Take 300 ml distilled water and add 225 9 potassium hydroxide (KOH) and dissolve.
Make up to 500 ml.
4.4. Nana indicator
Calcon carboxyl acid (C21H1402N25)
4.5. Hydrochloric acid (1+1)
Mix equal volumes of DM water and concentrated HC1.
4.6. Nitric acid (HNO3)
Concentrated nitric acid
5. STANDARDIZATION
Pipette 10.0 ml of the standard calcium solution (1 ml = 1 mg CaCO3) to 250 ml conical flask.
Add some 90 ml distilled water to it. Add 4 ml of calcium hardness buffer. Add a pinch of
Nana indicator and mix. Take the EDTA solution in the burette. Titrate the calcium solution
with EDTA from the burette, until the colour changes from reddish purple to blue. Note the
titre reading.
Normality of EDTA = _____2____
Titre reading
Adjust the normality of the EDTA solution by adding calculated amount of EDTA or diluting
with distilled water. The final titre reading should be 10.0 ml for the 10.0 ml calcium standard
solution.
6. PROCEDURE
6.1. Crush the sample using the electric grinder to a fine powder and dry the sample for
one hour at a temperature of 105 - 110 °C and cool.
6.2. Weigh ~0.5 gram of the sample to a 250 ml beaker, add a few drops of demin. water
and make a slurry of the powder. Note the weight as Wt.
6.3. Add 10 ml of hydrochloric acid solution (1+1), keep one glass rod inserted to the
beaker, heat to dissolve and dilute to 100 ml.
6.4. Add 10 - 15 drops of concentrated nitric acid and boil for 2 - 3 minutes with the
beaker covered with a watch glass.
6.5. Cool and transfer quantitatively to a 250 ml volumetric flask and make up to the mark
using demin. water.
6.6. Pipette 10.0 ml of the made up sample solution to a 250 ml conical flask. Add 50 ml
distilled water to it. Add 4 ml of calcium hardness buffer. Add a pinch of Nana
indicator and mix. Take the EOTA solution in the burette. Titrate the sample solution
with EDTA from the burette, until the colour changes from reddish purple to blue.
Note the titre reading as TR.
7. CALCULATIONS
% Calcium carbonate as CaCO3 = TR x 0.02 x 50 x 100

10 x 4 x Wt
Where,
TR = Titre reading of 0.02N EDTA
Wt = Weight of sample taken in grams.

APHA 2340 C & 3500Ca B
GLAB-57 (Revised) - Quality check of Lime stone Page 3 of 3
Page 223
SECTION : WATER
THE DETERMINATION OF SALINITY OF SOILS

(ELECTRICAL CONDUCTIVITY METHOD)

Page 224

SALINITY OF SOIL (ELECTRICAL CONDUCTIVITY METHOD)
G-LAB METHOD No.58
(By K.P. Abraham)
1. SCOPE
This test method covers the determination of the salinity of soils. Excessive soluble salts and
high exchangable sodium gives problems to soil fertility. On this basis the soils are divided
into three categories as 1. Saline soils,
2. Saline alkali soils and 3. Sodic soils.
2. PRINCIPLE
The electrical conductivity is directly proportional to the concentration of soluble salts in
soils and is measured as millimhos/cm in the saturation extract of soils. The critical limits of
electrical conductivity for soil salinity depends on
• Texture of the soil.

• Nature of the salt present.
• The plant species grown.
3. APPARATUS
3.1. Conductivity meter with probe. WTW Multiline 4
3.2. Thermometer, 0 – 50°C
3.3. Suction pump
3.4. Filtration assembly with buchner

funnel, suction flask and filter
paper.
3.5. Spatula
3.6. Volumetric flask, 100 ml.
3.7. Pipette, 5 ml.
4. REAGENT
GLAB-58 (Revised) - Salinity of Soil Page 2 of 4

Page 225
5. PROCEDURE
5.1. Soil sampling
5.1.1. Soil should be sampled from the active root zone of the plants, usually
from 6” below
5.1.2. Composite the sample for the average value

5.1.3. It is further air dried in the shade and sieved through a 2 mm standard
sieve.
5.2. Preparing saturated soil paste.

5.2.1. Fill the 250 ml beaker approximately to 100 ml mark with the soil sample.
5.2.2. Slowly add distilled water while stirring and mixing with the spatula until
the soil glistens and flows slightly when the beaker is tilted. Be certain that
the paste is smooth and without any dry or consolidated lumps.
5.2.3. Allow to stand covered for 4 hours.
5.3. Obtaining the saturated extract
5.3.1. Place a clean filter paper into the Buchner funnel
5.3.2. Transfer the saturated paste onto this filter paper. The paste should cover
the bottom of the buchner funnel.
5.3.3. Place the funnel on the suction flask and connect it to the suction pump.
5.3.4. Start the pump and collect the filtrate in the suction flask.
5.4. Measuring the electrical conductivity (EC) of the extract.
5.4.1. Pipette 5.0 ml of the saturated extract to a 100 ml volumetric flask. Dilute to
the mark with distilled water. Mix well and transfer this to a clean beaker.
5.4.2. Connect the conductivity probe to the meter.
5.4.3. Set the instrument to ON position. Change the mode to Conductivity
5.4.4. Immerse the conductivity probe into the sample. The conductivity meter will
now indicate the exact electrical conductivity in µmhos/cm. Record this
reading.
6. CALCULATION
Electrical conductivity mmhos/cm = Meter reading x 100 x _1__

5 1000
i.e. Soil salinity mmhos/cm = Meter reading x 0.02.

Page 226
7. RESULT
The electrical conductivities for soil salinity are out lined broadly in following table.
Electrical conductivity
No. Nature of the Soil
mmhos/cm at 25°C
1 0 Salinity effect mostly negligible

Yield of very sensitive crops
2 2
restricted.
3 4 Yield of many crops are restricted.
Only tolerant crops yeild
4 8
satisfactory.
5 16 Only a few tolerant crops survive.
Reference : Hand book of agriculture, No.60, USDA 1954.

Operation manual of WTW Multiline 4

Page 227
SECTION : WATER
THE DETERMINATION OF SALINITY, CHLOROSITY AND
CHLORINITY OF WATER (ARGENTOMETRIC METHOD)

Page 228

SALINITY, CHLOROSITY AND CHLORINITY OF WATER
(ARGENTOMETRIC METHOD)
G-LAB METHOD No.59
(By K.P. Abraham)
1. SCOPE
This method covers the determination of salinity, chlorosity and chlorinity of well waters,
industrial waters and sea water.
2. PRINCIPLE
Salinity is defined as the total solids in water after all carbonates have been converted to
oxides, all bromides and iodides have been replaced by chloride, and all organic matter has
been oxidised, It is numerically smaller than the total dissolved solids and usually is reported as
grams per kilogram. Chlorinity includes chloride, bromide and iodide all reported as chlorides
and chlorosity is the chlorinity multiplied by the water density at 20°C.
The chlorosity of the sample is determined by the titration of the sample with standard silver
nitrate solution using potassium chromate as indicator. The end point is indicated by
persistence of the brick-red silver chromate colour,
3. APPARATUS
3.1. Automatic burette, 25 ml.
3.3. Pipettes, 1, 10, & 25 ml.
3.5. Volumetric flask, 1 lit.
3.6. Indicator bottle
4. REAGENTS
4.1. Potassium chromate indicator solution
Dissolve 5 g of potassium chromate (K2CrO4) in 100 ml distilled water.
4.2. Silver nitrate solution, standard (0.141 N)

weight at 40°C. Dissolve 23.954 + 0.001 g of the crushed, dried, crystals in distilled
water and dilute to 11 with distilled water. Standardize with standard NaC1 solution
(0.141 N).
GLAB-59 (Revised) - Salinity, Chlorosity, and Chlorinity (Water) Page 2 of 3

Page 229
4.3. Sodium chloride solution, standard (0.141 M)
Dry about 10 g of sodium chloride (NaCl) for 1 hour at 140°C. Dissolve 8.2417 +
0.0002 g of the dry salt in distilled water and dilute to 11 at 25°C in a volumetric
flask.
5. STANDARDIZATION
Pipette 10.0 ml of the standard sodium chloride solution (0.141 M) in a 250 ml conical flask.
Add 5 drops of potassium chromate indicator and mix titrate against the silver nitrate solution
from a burette. Note the end point when the brick-red colour persists.
0.141M NaC1 = 0.141 N NaC1

Normality of AgNO3 = 10 x 0.141
Titre reading, ml
If the normality of the silver nitrate difference from 0.141, then adjust the normality to 0.141
by adding calculated volume of distilled water or calculate quantity of silver nitrate.
Standardize again, after the adjustments, with same procedure given earlier.
0.141 N AgNO3 = 0.141 N AgNO3
6. PROCEDURE
Take samples as follows:
100 ml if conductivity <700 µS/cm
50 ml if conductivity >700 but (1500 µS/cm
5 ml if the sample in a saline water (eg. sea water, brine recycle etc).
The remaining portion, distilled water to be added to make the volume about 100 ml. Add 5
drops of K2CrO4 indicator and mix. Titrate against the standard AgNO3 solution (0.141 N),
until the brick-red colour persists. Note the titre reading, ml.
7. CALCULATIONS
7.1 Chlorosity, g/litre = TR x N x 0.0355 x 1000
Sample volume. ml
Where,
TR = Titre reading, ml
N = Normality of AgNO3
7.2. Chlorinity, g/kg = chlorosity ÷ density of the sample at 20°C.
7.3. Salinity, g/Kg = 0.03 + 1.805 x (chlorinity, g/kg)
Reference : APHA 2520
GLAB-59 (Revised) - Salinity, Chlorosity, and Chlorinity (Water) Page 3 of 3

Page 230
SECTION : WATER
THE DETERMINATION OF SETTLED VOLUME

Page 231

SETTLED VOLUME (RAW SEWAGE)
G-LAB METHOD No.60
(By K.P. Abraham)
1. SCOPE
This method cover the determination sewage. This analysis is important waste water treatment
processes and of the settled volume of an untreated in the control of biological and physical
for assessing compliance.
2. PRINCIPLE
Settleable solids or sludge will settle down if it is allowed to settle by allowing the sample to
stand undisturbed. Here the sample is allowed to stand undisturbed in a measuring cylinder
for 30 minutes and the settled volume is noted.
3. APPARATUS
3.1. Settling column

1 litre graduated measuring cylinder
3.2. Gloves
3.3. Alarm clock
4. REAGENTS
4.2. Dettol
5. PROCEDURE
Mix the sample thoroughly in the sample bottle. Transfer 1 litre of the sample to the settling
column. Set the clock alarm for 30 minutes. Start the clock. Exactly after 30 minutes take the
volume of the settled sludge.
6. CALCULATIONS
Report the settled volume of the sludge in percentage as the settled volume of the sample for
30 minutes.
7. SAFETY NOTE
Dispose of the sample after analysis properly and clean the hands thoroughly with a dilute
dettol solution.
Reference APHA 2710 C
GLAB-60 (Revised) - Settled volume Page 2 of 2

Page 232
SECTION : WATER
THE DETERMINATION OF SILICA

Page 233

SILICA (COLORIMETRIC, MOLYBDATE-REACTIVE SILICA)
G-LAB METHOD No.61
(By K.P. Abraham)
1. SCOPE
This method covers the photometric determination of molybdate-reactive silica in water i.e.
drinking water, steam condensate, boiler water etc. Due to the complexity of silica chemistry,
the form of silica measured is defined by the analytical method as molybdate-reactive silica.
The useful range of this method is 20 to 100 µg/l.
2. PRINCIPLE
This method is based on the reaction of the soluble silica with molybdate ion to form a
greenish-yellow complex, which in turn is converted to a blue complex by reduction with
l-amino-2-naphthol-1-sulphonic acid (ANSA). Oxalic acid is added after the molybdate to
prevent interference by phosphates, forming phosphor-molybdate complexes, which are
reduced by ANSA in the same way.
Note: Colour and turbidity will interfere if not removed by filtration or dilution.
3. APPARATUS
3.1 Spectrophotometer.
JASCO, Model V 530
3.2 Sample cells: 10 mm & 50 mm.
3.3 Polythene bottles
3.4 Plastic cups, 250 ml, pipettes & volumetric flasks.
3.5 Pipette, polythene
3.6 Platinum crucible.
3.7 Volumetric flasks, ploythene, 100 ml
3.8 Muffle furnace.
3.9 Water polishing unit, Water -I
4. REAGENT
4.1 Silica Solution, Standard (1 ml = 1 mg SiO2)
Dissolve 4.73 g of sodium metasilicate (Na2SiO3.9H2O) in water and dilute to 1 L.

Check the concentration of this solution gravimetrically.
GLAB-61 (Revised) - Silica (Low range) Page 2 of 4

Page 234
4.2 Silica Solution, Standard (1 ml = 0.01 mg SiO2)
Pipet 10 ml of the standard silica solution (1 ml = 1 mg SiO2) to a 1 litre

volumetricflask and make upto the mark using deionised water. Store in polythene
bottle.
4.3 Silica Solution, Standard (1 ml = 1 µgSiO2)
Pipet 10 ml of the silica standard solution (1 ml = 0.01 mgSiO2) to a 100 ml

polythene volumetric flask and make up to 100 ml using deionised water.
4.4 Ammonium molybdate Solution
Dissolve 50 g ± 0.5 g of ammonium molybdate in about 400 ml of deionised water

at room temperature (solution 1). Add with caution 41.7 ml ± 0.5 ml of
concentrated sulphuric acid (H2SO4 Sp.gr.1.84) to 100 ml of distilled water and
allow to cool in a cold water bath (solution 2). Add solution 2 to solution 1 with
stirring. Cool and make up to 1 lit. Store in polythene bottle.
Note: This is stable for 3 months. If a blue colour appears, then it need not be
rejected until the reagent blank becomes unduly large i.e. >0.01 absorbance
using 50 mm cell.
4.5 Oxalic acid Solution
Dissolve 50 g of oxalic acid (C2H2O4.2H2O) in about 400 ml of deionised water and

make upto 500 ml. Store in polythene bottle. Stable for 3 months.
4.6 ANSA Reducing Agent
Dissolve 0.5 g of 1-amino-2-naphtol-4-sulphonic acid in 50 ml of a solution

containing 4 g of sodium sulphite (Na2SO3.7H2O). After dissolution, add the
solution to 150 ml of solution containing 20 g of sodium bisulphite (NaHSO3).or
18.27g of sodium disulphite (Na2S2O5).Store in plastic bottle and should be allowed
to stand for 1 hour before analysis.
5. PROCEDURE
5.1 Calibration and Standardization
Pipet 1.0, 2.0, 4.0, 6.0, 8.0 and 10.0 ml of the standard silica solution (1 ml = lµg
SiO2) to 250 ml plastic cups. Add remaining volume of silica free distilled water to
it to make the volume to 100 ml. This will give standard solutions with 0.01, 0.02,
0.04, 0.06, 0.08 and 0.10 mg/l SiO2. Take 50 ml each of these standards in plastic
cups and add 4 ml of ammonium molybdate solution to each and mix by swirling.
After 5 minutes add 5 ml of oxalic acid solution to each and mix by swirling.
Prepare the blank as follows: Take 50 ml DM water and add 5 ml of oxalic acid
solution and mix well. Then add 4 ml of ammonium molybdate solution and mix
well and wait for 5 minutes. After 1 minute add 2.0 ml of ANSA reducing agent to
each and mix by swirling. 10 minutes after the addition of the ANSA reagent,
measure the absorbance of each at 810 nm using 50 mm glass cells against blank
solution. Plot a graph of absorbance against concentration (mg/l SiO2). Calculate
the factor ‘F’ from the graph, where ‘F’ is mg/l SiO2 per unit absorbance.

Page 235
5.2 Sample Procedure
Take 50 ml sample and 50 ml distilled water as blank in 250 ml plastic cups. Follow
the same procedure used for the calibration. Note the sample absorbance.
In G - Station laboratory, by calling method No.5 in JASCO spectrophotometer, all
the parameters required for the silica determination (low range) will be set. The
concentration can be measured directly. For high range (0.1 to 1.0 ppm), use method
No.5 and use 10 mm cells for the determination.
6. CALCULATION
Silica, mg/l SiO2 = Sample absorbance x F
Ref: APHA 4500- SiO2

ASTM D859

Page 236
G – STATION LABORATORY
SECTION : WATER
THE DETERMINATION SILT DENSITY INDEX (SDI) OF WATER

Date issued : 20thOctober 2001
Page 237

SILT DENSITY INDEX (SDI) OF WATER
(Fouling Index or Pluggage Test)
G-LAB METHOD No.62
(By K.P. Abraham & V. Gengadurai)
1. SCOPE
This method covers the determination of the Silt Density Index (SDI) of water. This test
method can be used to indicate the quantity of particulate matter in water. Since the size, shape,
and nature of particulate matter in water tray vary, this test method is not an absolute
measurement of the quantity of particulate matter, The SDI has been empirically correlated
with the fouling tendency of sea water treatment equipment such as reverse osmosis devices,
2. PRINCIPLE
Water is passed through a 0.45 micron membrane filter at a constant applied gauge pressure of
30 psi (207 kPa), and the rate of pluggage of the filter is measured, The SDI is calculated from
the rate of pluggage. The determination is done manually in method A and this is done
automatically in method B.
METHOD - A
3. APPARATUS
3.1. SDI Assembly, as schematically described in Figure A.

3.2. Membrane filter, 47 mm diameter, gridded, and with a
mean pore size in the range 0.45 micron, inclusive.
3.3. Volumetric flask, 100 ml
3.4. Stop Match, graduated in hundredths of seconds
3.4. Thermometer, Liquid in glass, 0 - 100 °C
4. REAGENTS
4.1. None
5. CALIBRATION
5.1. None
6. PROCEDURE
6.1. Locate an appropriate sample point.

6.2. Hook up the SDI tester without a 0.45 micron filter in it.
6.3. Turn ON water the valve on the tester. Allow the water flow through the tester for 5
minutes.
6.4. Measure the temperature of the water sample and collect a sample for turbidity check and
for any other analysis, if required.
6.5. Close the valve on the tester and, using dull tweezers, place 0.45 micron white
Millipore filter (shiny side up) in the filter holder and gently tighten the thumb
nuts to hold it in the housing.
6.6. Partially open the valve and while water is flowing through the tester, slowly unscrew
GLAB-62 (Revised) - Silt Density Index (SDI) Page 2 of 5

Page 238
one or two of the thumb nuts and allow water to flood the tester housing and flow out of
the filter cavity to expel all air.
6.7. With the water still flowing out of the filter cavity, gently tighten the screws once you are
sure that there in no air entrapped in the filter housing. Open the valve fully and adjust
the pressure regulator so that the pressure reads 30 psi. Once the pressure is set, turn off
the valve.
6.8. Keep a 100 ml volumetric flask empty to collect the water sample.
6.9. Open the valve fully and immediately start a stopwatch and measure the time taken to
collect 100 ml sample in the volumetric flask, Record the time as T(0) seconds leaving
the valve open and the water flowing after the 100 ml mark has been reached.
6.10. Allow the water to keep flowing and, immediately at five minutes of elapsed test time,
measure the time in seconds it takes to fill the emptied 100 ml volumetric flask up to the
mark and record it as leaving the valve open and the water flowing, Repeat this
procedure at ten minutes and fifteen minutes of elapsed test times and record the times
as T(10) & T(15). After the time measurement has been made, the test can be stopped.
6.11. Measure the turbidity of the collected sample and record.
7. CALCULATIONS
If any time during the fifteen minutes test, you experience 100% pluggage (flow decreases to
slow drip), stop the test and calculate the Index as follows:
Silt Density Index, SDIT = ____% Pluggage___

Elapsed tine in Mints.
= __100____
Time (Min)
Where:
T= Elapsed Time
If the 15 minutes test is completed, calculate the Index as follows:
% Pluggage, % P30 = T(15) — T(0) X 100

T(15)
Silt Density Index, SDI15= % Pluggage_______

Elapsed time in Mints.
= % Pluggage = % P30
15 15
Where:
T(15) = Time in seconds after 15 minutes
T(0) = Time in seconds at start
Report the following information:
1. The SDI, with a subscript indicating the total elapsed tine (T) in minutes i.e; SDIT
2. The water temperature of the sample ° C
3. The manufacturer of the 0.45 micron membrane filter
Note:
For this test method, %P30 should not exceed 75%. If % P30 exceeds this value, use a shorter
time for T; that is, 5 or 10 minutes measurements.
Reference : a) Manual of SD1 Assembly
6) ASTM D 4189 - 95
Page 239

Page 240
METHOD - B
3. APPARATUS
3.1. Simple SDI Meter, Model :Simple SDI.

3.2. Membrane filter, 47 mm diameter,
gridded, and with a mean pore size in the
range 0.45 micron, inclusive.
3.3. Connecting hose.
4. REAGENTS
4.1. None
5. CALIBRATION
5.2. None
6. PROCEDURE
6.1. Connect Simple SDI to a water supply.

Use the strainer housing as a prefilter on all water supplies.
6.2. Open the membrane filter housing. Place the open housing over a bucket or sink.
Open the Inlet valve and flush the system for 10 seconds or until the line between
Simple SDI and the water source is clean.
6.3. Close the Inlet valve. Install a membrane filter in the membrane housing.
Partially close the housing.
6.4. Open the Inlet Valve slightly –just enough for water to flow from the sides of the filter
housing. Let the water flow until you see a clear bubble-free stream of water.
6.5. Close the Inlet Valve. Tighten the filter housing fully. Connect the drain line to the filter
housing.
6.6. Press POWER, and then ENTER on the controller to start the test.
6.7. Open the Inlet Valve.
Use the Pressure Regulator to adjust the water pressure to 30 psi.If the flow rate
increases
By 10% during the initial measurement an error message will appear and the test will
stop. Adjust the pressure to 30 psi and return to step 6.3.
6.8. Monitor the system for few seconds to ensure that the test has started normally.
6.9. Come back in 20 minutes for the results
7. CALCULATIONS
Results displayed as
SDI5 at 30psi.
SDI10 at 30psi.
SDI15 at 30psi.
Reference : Manual of simple SDI

Page 241
SECTION : WATER
THE DETERMINATION OF SODIUM CHLORITE CONTENT

Page 242

SODIUM CHLORITE (REDOX METHOD)
G-LAB METHOD No.63
(By K.P. Abraham)
1. SCOPE
This method covers the determination of the purity of sodium chlorite in bulk chemical
supplied for the Chlorine dioxide plant.
2. PRINCIPLE
The titrations are done automatically using titrant stored in a storage bottle, to the sample taken
in the titration vessel using an interchangeable burette operated by piston and stopcock. The
end point determination is done by the use of specific electrodes, combination platinum ring
electrode for redox titration. The dispensing and endpoint determinations are controlled by the
analytical conditions stored in the method, which is being used.
Automated iodometric titration with an excess of Sulphuric acid. This method is based on the
reduction action of the iodine ion on the chlorite species and on the subsequent determination
of iodine formed, by redox titration against Sodium thiosulphate; the potential step is located
around 230mV.
The METTLER DL 25 titrator is a complex analysis station for titrimetric analysis. This titrator
enable the titration to be performed and evaluated automatically. Considerations of the sample
weight in the result calculation allows the analysis to be reported in the desired unit. The setup
procedure for a particular analysis avoids the use of technical instrument parameters. The
titration can be carried out with optimum accuracy using just the standard configuration.
3. APPARATUS
3.1. Auto titrator METTLER DL 25

3.2. Combination platinum ring electrode
DG14O - SC
3.3. Titration vessel, glass, 250 ml
3.4. Pipettes, 2, 5, & 10 ml
3.7. Wash bottle
4. REAGENTS
4.1. Sulphuric acid, standard (0.5 mol/L)

water.
4.2. Sodium thiosulphate standard volumetric solution, 0.1 mol/L
Dissolve 24.8 g of Na2S2O3.5H2O in water. Add o.5 ml of Chloroform as preservative,
dilute to volume with water in a 1000 ml one-mark volumetric flask and mix
thoroughly.
4.3. Potassium iodide (Analar grade)
GLAB-63 (New) - Sodium chlorite (Autotitrator method) Page 2 of 4
Page 243
4.4. Potassium dichromate (Analar grade)

4.5. Starch solution (1%w/w)
4.6. Hydrochloric acid (1+1 by volume)
5. STANDARDIZATION
Weigh, to the nearest 0.1 mg (160±10) mg (m) of primary standard potassium dichromate into
a tared beaker. Place the contents of the beaker in a 500 ml stoppered conical flask, add 100 ml
of water and (2±0.5) g of potassium iodide and stir to dissolve. Add (15±1) ml of hydrochloric
acid solution (diluted 1+1 by volume), swirl, and allow to stand for 5 min. Titrate with the
sodium thiosulphate solution until the solution is pale yellow. Add (5±1) ml of starch solution
1% and titrate to the end point, i.e. to the disappearance of the blue-black colour. Record the
volume (v) used.
The concentration,c, of the sodium thiosulphate standard volumetric solution (Na2S2O3.5H2O),
expressed in moles per litre is given by the following equation:
c = _______m________
v x 49.0317
Where,
m = is the mass, in milligrams, of potassium dichromate (K2Cr2O7) weighed;
v = is the volume, in milliliters, of the sodium thiosulphate standard volumetric
solution used.
6. PROCEDURE
6.1. Switch ON the instrument and the printer.

6.2. Enter the DATE, MONTH, and YEAR and confirm each by pressing RUN.
6.3. Enter the method number as 10 and press METHOD.
6.4. Replace the storage bottle with the titrant, sodium thiosulphate 0.1mol/L.
6.5. Keep a wash beaker below the titration head.
6.6. Enter 15/30 and press BURET mL. This will complete the rinsing of the burette with
the titrant three times.
6.7. Clean the titration vessel and weight, to the nearest o.1mg, of the sample (~0.1 g), in
the titration vessel.
6.8. Add 300 ml of DM water to the titration vessel and 4g of potassium iodide and add,
with stirring 20ml of sulphuric acid (0.5mol/l).
6.9. Remove the wash beaker and connect the titration vessel.
6.10. Press RUN.
6.11. Enter the weight of the sample, in grams, and press RUN.
6.12. Enter the sample identity number as follows and press RUN.
0000 — 0999 — Non routine samples
*000 — *999 — All other methods (* Method No.)
6.13. When the BUSY light is blinking press RUN.
6.14. Wait for the titration to finish.
6.15. The result will be displayed as well as printed. Additional information and other
titration details can be printed by using the code operations.
6.16. After completing the analysis, replace the titrant with DM water/solvent.
6.17. Carry out the rinsing as in step No.6.6.
6.18. Replace the titration vessel with the wash beaker filled with DM water.
6.19. Switch OFF the instrument.

Page 244
7. CALCULATIONS
The sodium chlorite (NaClO2) content, C1, expressed as a percentage by mass (%w/w), is given
by the following equation. Assume 90.44 g of sodium chlorite (NaClO2) is equivalent to 1000
ml of sodium thiosulphate (Na2S2O3.5H2O = 0.1 mol/L).
C1 = _V1 x c x 2.262
m
where,
V1 = is the volume, in milliliters, of sodium thiosulphate standard volumetric
solution used for the sample titration to the endpoint.
c = is the concentration, moles per litre, of the sodium thiosulphate standard
volumetric solution.
m = is the weight in grams of the test sample
When the METTLER DL 25 auto titrator in G Station is used with the method # 10, result in
the specified unit along with the analytical parameters are printed out as a report after
completing the analysis by the titrator. Additional informations, titration curve, first derivative
of the titration curve, table of measured values of the titration curve also can be printed out
from the stored data.
Reference: Analysis method supplied by CAFFARO for Biocaf

Manual for METTLER DL 25 Titrator

Page 245
SECTION WATER
THE DETERMINATION OF SODIUM SULPHITE CONTENT

Page 246

SODIUM SULPHITE
G-LAB METHOD No.64
(By Philip Varkey)
1. SCOPE
This method covers the determination of sodium Sulphite content in Sodium Sulphite sample.
2. PRINCIPLE
A known excess of standard iodine solution is added to the material. The excess iodine solution
is back titrated with thiosulphite solution. The iodine consumed in oxidizing the sulphite to
sulphate gives a measure of the sulphite.
3. APPARATUS
3.1. Burette
3.2. Pipette
3.3. Conical flask
4. REAGENTS
4.1. Standard Iodine 0.10 N
4.2. Standard Thiosulphite 0.10 N
4.3. Starch
4.4. Dil. .HCl
5. REAGENT PREPERATION
5.1. Sodium Thiosulphate (0.10 N)

Dissolved 24.82 g of sodium Thiosulphate (Na2S2O3.5H20 ) in boiler and cooled
distilled water. Dilute to 1 L and add 0.10 g of sodium carbonate as a preservative.
This solution deteriorates on standing and should be standardized every week, using
standard Iodine solution.
5.2. Iodine solution, standard (0.05M)
quantitatively to a 1 Liter volumetric flask and make up to the mark with distilled
water.
0.50 g of soluble starch is made to a thin paste with a little cold water and then add 25
ml ofboiling water. Boil until a clean solution is obtained. This should be freshly
prepared as required.
5.4. Dilute HCl
GLAB-64 (New) - Sodium Sulphite (Iodimetric method) Page 2 of 3

Page 247
6. PROCEDURE
Weight accurately about 0.25 g of the material in a tared weighting bottle and
add it to exactly 50 ml of standard iodine solution acidified with 2.0 ml of
dilute hydrochloric acid. Allow to stand for 5 minutes and titrate the excess of
iodine with standard sodium thiosulphate solution using starch solution as
indicator, the end point of the reaction being indicated by the disappearance of
blue colour. Carry out a blank titration without using the material.
7. CALCULATION
Sodium Sulphite ( Na2SO3) content percent by mass
= 6.303 ( V1 – V 2 ) N
M
Where,
V1 = Volume in ml of standard sodium thiosulphate solution used in
the blank titration
V2 = Volume in ml of standard sodium thiosulphate solution used with
the material
N = Normality of standard sodium thiosulphate solution
M = Mass in gram of the material taken for the test
GLAB-64 (New) - Sodium Sulphite (Iodimetric method) Page 3 of 3

Page 248
SECTION : WATER
SPECIFIC GRAVITY/DENSITY OF LIQUIDS

Page 249

SPECIFIC GRAVITY/DENSITY OF LIQUIDS
G-LAB METHOD No.65
(By K.P. Abraham)
1. SCOPE
Different chemicals used in the plant to be checked for its physical properties to certify the
quality of it. Specific gravity / Density is one among them.
2. PRINCIPLE
The plumment supplied with the balance is immersed in the sample. While immersing into the
given liquid the plummet becomes lighter by buoyancy. The difference in weight is noted in the
balance as density.
3. APPARATUS
3.1. Density balance
3.2. Measuring jar
3.4. Thermometer, 0 to 50 °C
4. REAGENTS
4.1. Distilled water Distilled water at 20°C.
5. CALIBRATION
5.1. Zero point calibration

Carefully clean and dry the plummet and hang it on the hook. Keep both the sliding
weights on the 0.000 positions. Keep the beam weight in position. Turn the screws on
the foot and at the left of the beam to zero the balance.
6. PROCEDURE
6.1. Fill up the measuring jar 20 mm below the rim with the sample and immerse the
plummet into it.
6.2. Adjust the sliding weight and make the balance steady. If the density of the standard
is more than 1 then remove beam weight to steady the balance.
6.3. Read the weight by adding the slider weights. Add 1 if the hanging weight is off the
beam.
GLAB-65 (Revised) - Specific gravity or Density of liquids (Density balance method) Page 2 of 3
Page 250
6.4. The reading of the balance is corrected according to the correction schedule because,
air buoyancy and liquid bulge at the wire of the plummet, are going into the
measurement.
CORRECTION SCHEDULE
Density Correction Density Correction
0.6000 +0.0003 1.4000 -0.0006

0.7000 +0.0002 1.5000 -0.0008
0.8000 +0.0001 1.6000 -0.0010
0.9000 0.0000 1.7000 -0.0011
1.0000 0.0000 1.8000 -0.0013
1.1000 - 0.0002 1.9000 -0.0015
1.2000 - 0.0003 2.0000 -0.0017
1.3000 - 0.0005
7. CALCULATION
Density of the liquid = Sum of the slider weights + Correction, g/cc

(If the beam weight is on the beam)
= 1 +Sum of the slider weights + Correction, g/cc

(If the beam weight is off the beam)
GLAB-65 (Revised) - Specific gravity or Density of liquids (Density balance method) Page 3 of 3
Page 251
SECTION : WATER
THE DETERMINATION OF SULPHATE
(GRAVIMETRIC)

Page 252

SULPHATE BY GRAVIMETRIC METHOD
G-LAB METHOD No.66
(By K.P. Abraham)
1. SCOPE
This method covers the determination of sulphate ion in water and waste water. The
determination of sulphate, in the presence of sodium and magnesium, is required to avoid the
action of this ion when present in excess in drinking water.
2. PRINCIPLE
Sulphate is precipitated in a hydrochloric acid (HC1) solution as barium sulphate (BaSO4) by
the addition of barium chloride (BaCl) after the removal of insoluble matter. The precipitate is
filtered, washed, ignited and weighed as BaSO4, if present.
3. APPARATUS
3.1. Drying oven, equipped with thermostatic control.

3.2. Muffle furnace, with temperature indicator.
3.3. Hot plate.
3.4. Dessicator.
3.5. Filter papers, ashless No.42 and 41.
3.6. Funnel.
3.7. Platinum crucible, 50 ml.
3.9 Glass rod.
3.10. Watch glass.
3.11. Police man
3.12. Wash bottle.
4. REAGENTS
4.1. Barium chloride solution (118 g/l)

Dissolve 118 g of barium chloride (BaC12.2H2O) in water and dilute to 1 litre.
4.2. Hydrochloric acid (1+9)
Mix 1 volume of hydrochloric acid (HC1 sp.gr 1.19) with 9 volumes of water.
4.3. Hydrofluoric acid (48 to 51%)
Concentrated hydrofluric acid (HF)
4.4. Methyl orange indicator solution (0.5 g/l)
Dissolve 0.05 g of methyl orange in water and dilute to 100 ml.
4.5. Silver nitrate solution (l00g/l)
Dissolve 10 g of silver nitrate (AgNO3) in water and dilute to 100 ml.
4.6. Ammonium hydroxide (Sp.gr 0.90)
Concentrated ammonium hydroxide (NH4OH).
4.7. Sulphuric acid (Sp.gr 1.84)
Concentrated sulphuric acid (H2S04)
GLAB-66 (Revised) - Sulphate (Gravimetric method) Page 2 of 3

Page 253
5. PROCEDURE
Filter the sample if it is turbid, using a fine ashless filter paper (No.41). Wash the beaker and
the filter thoroughly with hot water.
Measure into the beaker a quantity of the clear sample containing sulphate ion equivalent to
10 to 50 mg of barium sulphate (BaSO4). Adjust the volume by evaporation or dilution with
distilled water to approximately 200 ml. Adjust the acidity of the sample to the methyl orange
end point using either ammonium hydroxide or hydrochloric acid. Then add 10 ml excess of
hydrochloric acid HC1
(1+9).
Heat the acidified solution to boiling and slowly add to it 5 ml of hot Barium chloride BaC12
solution, Keep the temperature just below boiling until the liquid has become clear and the
precipitate has settled out completely. In no case shall this settling period be less than 2 hours.
Filter the suspension of Barium Sulphate BaSO4 on an ashless filter paper (No.42) and wash
the precipitate with hot distilled water until the washings are free of chlorides, as indicated by
testing the last portion of the washings with Silver nitrate AgNO3 solution. Avoid excessive
washing.
Dry the filter paper with funnel in the drying oven. Then place the filter paper and contents in a
weighed platinum crucible, char and consume the paper slowly without flaming. Ignite the
residue at approximately 800°C for 1 hour, or until it is apparent that all the carbon has been
consumed.
Add a drop of Sulphuric acid H2S04 and a few drops of Hydrofluoric acid HF, and evaporate
under a fume hood to expel silica as silicon tetrafluoride (SIF4). Reignite at about 800°C, cool
in a desiccator, and weigh the BaS04
6. CALCULATION
Sulphate mg/l (ppm) = mg BaSO4 x 411.6

sample volume, ml
Reference: APHA 4500 S042- (B)
GLAB-66 (Revised) - Sulphate (Gravimetric method) Page 3 of 3

Page 254
SECT ION : WATER
DETERMINATION OF SULPHATE (SO4 --) ION
(TURBIDIMETRIC METHOD)

Page 255

SULPHATE (SO4--) (TURBIDIMETRIC METHOD)
G-LAB METHOD No.67
(By K.P. Abraham)
1 SCOPE
This turbidimetric test method covers the determination of sulphate ion in vaster in the range
from 1 to 40 mg/L of sulphate (SO4--) Sulphate ion if present more than 260 mg/L in drinking
water is reported to be causing cathartic act ion (especially in children) in the presence of
sodium and magnesium and also gives bad taste to the water.
2. PRINCIPLE
2.1 Sulphate ion is converted to a barium sulphate suspension under controlled

conditions. Sodium chloride and hydrochloric acid are added before precipitation to
inhibit the growth of microcrystals of barium sulphate. The optimum pH is
maintained to minimise the effect of variable amounts of other electrolytes present in
the sample upon the size of the suspended barium sulphate particles. The glycerol
solution added helps to stabilize the turbidity. The reaction vessel is shaken at the
same fashion as the standard solutions. The resulting turbidity is determined by a
nephelometer or a spectrophotometer at 420 nm wavelength.
2.2 Interferences:
Insoluble suspended matter in the sample must be removed. Dark colours that
cannot be compensated for in the procedure interfere with the measurement
of suspended barium Sulphate (BaSO4). Polyphosphates as low as 1 mg/L,
phosphonates in lower concentrations depending on the type and chlorides
>5000 mg/L will all cause negative interferences.
Aluminum, polymers and large quantities of organic material will cause non
uniform barium sulphate precipitations.
Silica >500 mg,/L may cause positive interference.
In the presence of organic matter sulphates are reduced to sulphide by
sulphate reducing bacteria. In order to prevent this (negative interference)
sample is to be stored at 4°C
The formation of barium sulphate suspension is very critical so it is essential
to adhere rigidly to the experimental procedures detailed below.
3. APPARATUS
3.1 Spectrophotometer JASCO model V 530 or Filter

photometer model MPM 1500 with 420 nm filter.
3.2 10 mm cuvette cell.
3.3 Filteration apparatus.
3.4 Pipettes, volumetric flasks, conical flasks etc.
3.5 Stop watch.
3.8 Magnetic stirring apparatus.
GLAB-67 (Revised) - Sulphate (Turbidimetric method) Page 2 of 4

Page 256
4 REAGENTS
4.1 Glycerine (1+1)

Mix 500 ml glycerine with 500ml of demin water
4.2 Sodium chloride solution (24%)
Dissolve 240 grams of NaCl in demin water containing 20 ml of 35% hydrochloric
acid and dilute to 1 litre with demin water-
4.3 Barium chloride solution solution 6%
Disslove 60 grams of BaCl2 .2H2O in 1litre demin water.
4.4 Sulphate standard solution 100 mg/L (1ml = 0.100 mg SO4--)
Dissolve 0.1479 grams of anhydrous sodium sulphate (Na2SO4) in demin water and
make up 1 litre or take 10.0 ml of standard 1000 mg/L (ready made) sulphate
solution and make up to100 ml with demin water.
5 CALIBRATION
Follow the procedures given in section 6.3 onwards (and the specific procedures of the
instruments used) using the standards prepared as follows. Take 0.0,1.0, 2.5, 5.0, 7.5,
10.0, 5.0, 20.0,30.0 and 40.0 mg/L respectively.
Plot a graph using concentration versus absorbance and calculating the factor F for unit
absorbance.
6 PROCEDURE
6.1 Filter the sample if it is turbid and bring it to room temperature (15 to 30°C)
6.2 Measure 50 ml or less of the sample containing sulphate between 0.5 and 4.0 mg and
dilute to 50 ml with demin water, if required. (Note: due to the solubility problems of
BaSO4, concentrations of sulphate below 5 mg/L is determined by adding 5 ml of the
standard sulphate (0.5 mg sulphate) into the sample which. must have to be subtracted
from the final result).
6.3 Keep the sample over the magnetic stirrer and start mixing at a uniform speed.
6.4 Add 10 ml of 1+1 glycerine into the sample.
6.5 Add 5 ml of 24% sodium chloride solution.
6.4 Add 5 ml of 6% barium chloride.
6.5 Now stir for 1 minute, allow to stand for 4 minutes undisturbed and remix for 15
seconds.
Note: The stirring should be at a constant rate in all the determinations.
6.8 Immediately after stirring measure the absorbance using the spectrophotometer
JASCO Model V 530 or filter photometer MPM 1500 at 420 nm wave length.
(Follow the specific instrument’s operation procedures for measurements and the
determination of the concentrations).
6.9 Blank:To compensate any colour or turbidity of the sample a blank is carried out in
the manner using the sample and without the addition of barium chloride.
7.0 If any interference are suspected dilute the sample with equal volume of water and
determine the sulphate concentration again. If the result is half of the original sample
means no interferences.

Page 257
7 CALCULATION
If JASCO Spectrophotometer is used read the concentration mg/L as SO4 directly.
If Filter photometer MPM 1500 is used

Concentration of SO4, mg/L = Absorbance X F
References: APHA 4500- SO4-- E

ASTM D 516

Page 258
SECTION : WATER
THE DETERMINATION OF TOTAL SUSPENDED SOLIDS

Page 259

TOTAL SUSPENDED SOLIDS DRIED AT 103 – 105°C
G-LAB METHOD No.68
(By K.P. Abraham)
1. SCOPE
This method covers the determination of suspended particles in all types of waters i.e., cooling
water, sewage effluents, seawater, industrial wastewater etc.
2. PRINCIPLE
A well mixed sample is filtered through a weighed membrane filter paper (0.45µm) and the
residue retained on the filter is dried to a constant weight at 103°C to l050C. The increase in
weight of the filter represents the total suspended solids.
3. APPARATUS
3.1. Millipore filtration unit, which can hold 47 mm diameter filter paper.
3.2. Membrane filter, Whatman 47 mm dia and 0.45µm pore size, WCN type.
3.3. Desiccators
3.5. Drying oven 1050C
3.6. Measuring cylinder
4. REAGENTS
5. PROCEDURE
Place the membrane filter paper (0.45 µm) in the filtration unit. Using slight suction, wash the
filter with about 100 ml distilled water and when free from excess water carefully remove the
paper, place on a watch glass and heat in an over for 1 hour at 105 0C. Cool the filter paper in a
desiccator and weigh to the nearest 0.1 mg. Let the weight be W1 mg. Replace the paper in the
filtration unit and moisten with water. Measure a suitable volume V ml of the sample which
yields not more than 200 mg residue, after the sample has been well mixed by rapid shaking to
disperse the suspended matter. Filter under slight suction ensuring that all solids are transferred
to the filter paper by washing down with distilled water. Wash the residue three times with 5 to
10 ml of water allowing it to drain free from water after each wash. Carefully remove the filter
paper, place on a watch glass and dry it in an oven at 1050C for 1 hour. Allow to cool in a
desiccator and weigh the paper. Let the weight be W2 mg. Check for substantially constant
weight by further heating for 15 min. and cooling.
GLAB-68 (Revised) - Suspended solids Page 2 of 3

Page 260
6. CALCULATION
Suspended solids mg/l = (W2—W1) x 1000

V
Where,
W1 = Wt. of filter paper’ mg
W2 = Wt. of filter paper + residue, mg
V = Volume of sample, ml
Reference : APHA 2540 D

Analysis of raw, potable and waste waters, H.M.S.O.
Standard methods for the examination of water and waste water.
GLAB-68 (Revised) - Suspended solids Page 3 of 3

Page 261
SECTION : WATER
THE DETERMINATION OF TANNIN

Page 262

TANNIN IN AIR CONDITION SYSTEM COOLING WATER
G-LAB METHOD No.69
(By K.P. Abraham)
1. SCOPE
This method covers the determination of tannin in Air Condition system cooling water systems.
This could cover a range from 10 to 250 µg of tannin in 25 ml.
2. PRINCIPLE
Tannin and other hydroxylated aromatic compounds reduce tungstophosphoric and

molybdophosphoric acids to produce a water soluble blue complex, the concentration of which
is measured spectrophotometrically. The wavelength used is 700 nm.
Interferences.
Under the conditions described in the method the presence separately, in quantities indicated
of:
Calcium (Ca2+) 25 mg
Chloride (Cl- ) 25 mg
Iron (Fe2+) 5 µg
Iron (Fe3+) 250 µg
Magnesium (Mg2+) 25 mg
Nitrite (NO2- ) 1 mg
Silicate (SiO3)2- 1 mg
Sulphate (SO4)2- 25 mg
Sulphite (SO3)2- 5 µg
in the test solution do not cause significant interference. Sulphide ions (s-), phenols and cresols
interfere and shall be absent.
3. APPARATUS
JASCO Model V 530
3.2. Glass cells, 10 mm.
3.3. Pipettes, 1 ml, 2 ml, 20 ml.
3.4. Volumetric flasks, 50 ml (10 Nos.), 1 lit.
4. REAGENTS
4.1. Sodium carbonate 150g/l reagent solution. Filter if necessary, before use.
4.2. Sodium hexametaphosphate. 250 g/l reagent solution.

4.3. Tungsto-molybdophosphoric acid reagent. Dissolve 100 g of sodium tungstate
dihydrate, 10 g of dodeca—molybdophosphoric acid and 50 ml of concentrated
orthophosphoric acid, 90% (m/m) (43N) in 750 ml of water. Boil the liquid gently
GLAB-69 (Revised) - Tannin Page 2 of 3

Page 263
under relfux for 2 h, cool and make up to 1000 ml with water.

4.4. Tannin standard solutions. Where tannin is known to have been added to the water
from which the sample was taken, use a sample of this tannin to prepare these
solutions.
a. Dissolve 1.00 g of the tannin in about 80 ml of water and make up to the
mark in a l000ml one mark volumetric flask.
b. Tannin working solution. Pipette 10.00 ml of the tannin stock solution into a
1000 ml one-mark volumetric flask and make up to the mark with water.
Mix. Prepare freshly as required.
l ml = 10 µg of tannin.
5. CALIBRATION
Into each of a series of 50 ml stoppered cylinders introduce accurately measured volumes of

the tannin working solution corresponding to 0, 25, 50, 100, 150, 200 and 250pg of tannin,
make up to 25 ml with water and mix. To each cylinder add 1 ml of the tungsto-
molybdophosphoric acid reagent and mix. Add 2 ml of the sodium hexametaphosphate
solution, mix and allow the mixture to stand for 5 min. Add 20 ml of the sodium carbonate
solution, mix and make up the volume to the mark with distilled water and allow to stand for a
further 10 min.
Measure the absorbance of each solution in a spectrophotometer at a known temperature

between 200C and 25°C at a wavelength 700 nm using a 10 mm cell. Measure the absorbance
against the blank with no added tannin. Plot a calibration graph of absorbance against mg/l
tannin. Assuming the sample volume of 50 ml, the above standard solutions will correspond to
0.5, 1.0, 2.0, 3.0, 4.0 and 5.0 mg/l tannin. Calculate a factor, which is equal to mg/l tannin,
which produces unit absorbance.
6. PROCEDURE
Measure a volume of the sample (up to 25 ml) that will contain not more than 250µg of tannin
into a 50 ml stoppered cylinder and 25 ml of water into a second 50 ml stoppered cylinder.
Make up the volume of sample to 25 ml, if necessary. Mix the contents of each cylinder and
add to each 1 ml of the tungsto—molybdophosphoric acid solution and mix. Add 2 ml of the
sodium hexametaphosphate solution mix and allow to stand for 5 min. Add 20 ml of the
sodium carbonate solution mix and allow to stand for further 10 min.
Measure the absorbance of the sample solution against the blank solution using 10 mm cells in
the spectrophotometer at a temperature within l°C of that at which the calibration graph was
prepared and at the same wavelength. Note the absorbance.
7. CALCULATION
Tannin, mg/litre = Absorbance x F x D
Where,
Absorbance = Sample absorbance against the blank
F = Factor from the calibration graph
D = Dilution factor, ______50_______
Sample volume, ml
Reference : APHA 5550 B
GLAB-69 (Revised) - Tannin Page 3 of 3

Page 264
SECTION : WATER
THE DETERMINATION OP TOTAL DISSOLVED SOLIDS

Page 265

TOTAL DISSOLVED SOLIDS AT 180°C
G-LAB METHOD No.70
(By K.P. Abraham)
1. SCOPE
This method is primarily applicable to water that will yield a residue on evaporation and later
drying at 180 0C. This is applicable to drinking water, cooling water, sea water etc.
2. PRINCIPLE
A well mixed sample is filtered through a fine filter paper (No. 41) and the filtrate is
evaporated to dryness in a weighed dish and dried to constant weight at 180°C. The increase
in dish weight represents the total dissolved solids.
3. APPARATUS
3.1 Evaporating dishes.

200 ml, platinum dishes or porcelain dishes.
3.2 Drying oven, for operation at 180 ± 2°C.
3.3 Steam bath.
3.4 Desiccators
3.5 Analytical balance, capable of weighing to 0.1 mg.
3.6 Filtration apparatus.
4. REAGENTS
4.1 Distilled water.
5. PROCEDURE
Mix the sample well and filter it through a fine filter paper. Heat the clean platinum/porcelain
dish to 180 ± 2°C for 1 hour in an oven. Cool it in a desiccator and take the weight. Measure a
certain volume of the filtered sample to yield between 5 and 200 mg dried residue, to the dish.
Evaporate to dryness on a steam bath. If the sample volume exceeds dish capacity add
successive portions to the same dish after evaporation. Dry for at least 1 hour in an oven at
180 ± 2°C, cool in a desiccator and weigh. Repeat drying cycle of drying, cooling, desiccating
and weighing until a constant weight is obtained or until weight loss is less than 4% of
previous weight or 0.5 mg, whichever is less.
For brine samples,

Sample volume = 50 ml
Drying time at 180 0C = 4 hrs.
GLAB-70 (Revised) - Total Dissolved Solids at 180°C Page 2 of 3

Page 266
6. CALCULATION
Total dissolved solids, mg/l = _(A—B) X 1000__

Sample volume, ml
Where,
A = weight of dried residue + dish in mg.
B = weight of dish, mg.
Reference : APHA 2540 C
GLAB-70 (Revised) - Total Dissolved Solids at 180°C Page 3 of 3

Page 267
G – STATION LABORATORY
SECTION : WATER
THE DETERMINATION OF TOTAL KJELDAHL NITROGEN

Date issued : 21st July 1999
Page 268

TOTAL KJELDAHL NITROGEN IN AQUEOUS LIQUIDS
(Digesdahl Digestion Method)
G-LAB METHOD No.71
(By K.P. Abraham)
1. SCOPE
This method covers the determination of Total Kjelhahl Nitrogen (TKN) in sewage water.
Using this method TKN concentration varying from 1 to 200 mg/L can be determined. For
higher concentrations use proportionate dilute sample.
2. PRINCIPLE
Analysing the Kjeldahl Nitrogen consists of two principle process. The first process is the
sample digestion, which is the oxidation of organic matter arid conversion of the organic
nitrogen and ammonia nitrogen content to Ammonium sulphate. The second process,
Ammonium sulphate measurement, determines the amount of ammonia in the digest using
Nesslerisation method.
Hydrogen peroxide is added to the sample, which is digested with Sulphuric acid at 440°C.
This converts the Kjeldahl Nitrogen to Ammonium sulphate.
The carbonisation period prior to the addition of Hydrogen peroxide provides a reducing
environment, which helps convert, organic nitrogen to ammonia. In the presence of oxidizable
carbon compounds, Sulphuric acid reacts to produce Sulphur dioxide, which is an active
reducing agent.
H2S04 → H20 + SO2 + ½ O2
During oxidation time, Hydrogen peroxide reacts immediately with H2SO4 at digestion
temperature to give H2SO5 (Peroxymonosulphuric acid).
H2S04 + H2O2 → H2S05 + H2O
This is an extremely powerful oxidizing agent towards carbonaceous material. An aliquot of

the digest is neutralized and then Nesslerized in the presence of metal stabilizer and a
dispersing agent and the nitrogen content is determined colourimetrically.
3. APPARATUS
3.1. Digesdahl Digestion Apparatus, HACH Model 23130 - 21
3.2. Dispenser, Pour-out, 10 ml
3.3. Pipette, 10 ml
3.4. TenSette pipette, 0.1 to 1.0 ml
3.5. Pipette filler, Safety bulb
3.6. Boiling chips, Silicon carbide.
3.7. Safety glasses
3.9. Beaker, 500 ml & 250 ml
3.10. Bottle, Wash bottle, 1L
3.11. Bulb, Dropper bulb, 10 ml
3.12. Flask, Flat bottom volumetric flask, 100 ml
GLAB-71 (Revised) - Total Kjeldahl Nitrogen Page 2 of 4
Page 269
3.13. Cylinder, Graduated, 50 ml

3.14. Spatula, Stainless steel, 10 cm
3.15. Cots, Finger cots
3.16. Spectrophotometer, HACH Model DR 2010
4. REAGENTS
4.1. Hydrogen peroxide 30 %, HACH Cat.No.144 - 45

4.2. Sulphuric acid, Sp.gravity 1.84, HACH Cat.No.979 - 09
4.3. TKN indicator solution, HACH Cat.No.22519 - 26
4.4. Potassium hydroxide lN, HACH Cat.No.23144 - 26
4.5. Potassium hydroxide 8N, HACH Cat.No.282-49
4.6. Mineral Stabilizer, HACH Cat.No.23766 - 26
4.7. Dispersing agent, polyvinyl alcohol, HACH Cat.No.23765 - 26
4.8. Nessler’s reagent, HACH Cat.No.21l94 - 49
4.9. Sodium thiosulphate solution 0.1 N, HACH Cat.No.323 - 37
4.10. Nitrogen, Ammonia standard solution, 50 mg/L NH3 - N, HACH Cat. No. 14791 - 10
5. CALIBRATION
Take 7 Nos. of 100 ml volumetric flask. Pipette 10 ml DM water each to all of then. Pipette 0,
0.2, 0.4, 0.8, 1.2, 1.6, and 2.0 ml of standard ammonia solution (50 mg/L NH3 - N). Add 3
drops of TKN indicator and mix. Add 2 drops of 1N KOH and mix. Add 6 drops of dispersing
agent and then make up to 100 ml with DM water. This will provide 0, 0.1, 0.2, 0.4, 0.6, 0.8 &
1.0 mg/L NH3 - N. Shake well and then add 1 ml each of Nessler’s reagent to each and mix
well. Wait 30 minutes for colour development. Measure the absorbance at 430 nm wavelength
using 50 mm cell. Use the first solution i.e; DM water with the reagents as the blank. Plot a
calibration graph with absorbance against mg/L of NH3 - N. Calculate the factor ‘F’ which is
equal to the mg/L NH3 - N for unit absorbance.
6. PROCEDURE
6.1. Transfer 20 ml sample into a 100 ml Digesdahl digestion flask. In samples with more
than 1% solids present, take sample volume = 40 + % solids. Add remaining volume of
DM water to make the volume to 40 ml.
6.2. Add carefully 3 ml of concentrated Sulphuric acid (Sp.gravity 1.84)
6.3. Add 3 boiling chips which is pretreated with 1:1 Nitric acid.
6.4. Switch ON the heater and set the temperature to 440°C. When the proper temperature
is reached turn ON the water to the aspirator and make sure there is suction to the
fractionating column.
6.5. Place the flask weight followed by the fractionating column with the funnel on the
flask. Place the flask on the heater and heat until the Sulphuric acid reflux. It takes
about 14 minutes.
6.6. Boil for 4 more minutes.
Note: Do not boil the sample to dryness. If Sulphuric acid is not present in the
flask after 4 minutes heating, do not proceed. Discard it and start over again from the
beginning.
6.7. Add 10 ml of 30% Hydrogen peroxide to the charred sample via the funnel on the
fractionating column.
Note: If the digest does not turn colourless, add 5 ml. increments of Hydrogen
peroxide until the digest becomes clear or does not change colour.
6.8. After addition of Hydrogen peroxide is complete boil off excess Hydrogen peroxide
by heating for one more minute.
Note: Do not heat to dryness.
Page 270
6.9. Take the hot flask off the heater using the finger cots and allow the flask to cool.
6.10. Remove the fractionating column from the digestion flask.
6.11. Dilute the digest to 100 ml mark with deionised water. Invert several times to mix.
Note: Add deionised water slowly at first. Cool the flask, if necessary for handling.
6.12. Run a blank, the same way, with 40 ml DM water and make up to 100 ml.
6.13. Pipette 10 ml sample and 10 ml blank solution to 100 ml volumetric flasks.
6.14. Add 3 drops of TKN indicator and mix.
6.15. Add SN 1(011 solution drop wise and shake, till the colour changes to pale blue ( 30
drops)
6.16. Add 2 drops of 8N KOH solution to each.
6.17. Add 6 drops of metal stabilizer and shake well.
6.18. Add 3 drops of dispersing agent and shake well.
6.19. Make up to 100 ml with DM water.
6.20. Add 1 ml Nessler’s reagent (HACH) and mix well and wait for 30 minutes for the
colour development.
6.21. Prepare the blank water also the same way as the sample.
6.22. Measure the absorbance of the sample using 50 mm cell at 430 nm wavelength
against the blank water.
6.23. Note the absorbance reading.
6.24. Concentration can directly measured using method no.37 in the G - Station laboratory
HACH spectrophotometer. Make instrument zero with blank water with the reagents.
7. CALCULATIONS
Total Kjeldahl Nitrogen, mg/l = Absorbance x F x 50
Where
F = Factor from the calibration graph
50 = No. of dilutions
Reference: Manual of MACH Digesdahl Digestion Apparatus Water Analysis hand

book (HACH)

Page 271
SECTION WATER
THE DETERMINATION OF TOTAL NITROGEN
(TNT Persulfate Digestion Method )
Method No. : GLAB -72

Page 272

TOTAL NITROGEN
[TNT Persulfate Digestion Method ]
G-LAB METHOD No.72
(By Philip Varkey)
1. SCOPE
This method covers the determination of Total Nitrogen in sewage water, using this method
Total Nitrogen concentration varying from 0 to 25 mg/l can be determined. For higher
concentration use proportionate dilute sample.
2. PRINCIPLE
An alkaline persulphate digestion converts all forms of nitrogen to nitrate. Sodium

metabisulphite is added after the digestion to eliminate halide interference. Nitrate then reacts
with chromotropic acid under strong acidic conditions to form a yellow complex with an
absorbance maximum at 410 nm.
3. APPARATUS
3.1. COD Reactor

3.2. Spectrophotometer – HACH 2010
3.3. Pipette
4. REAGENTS
4.1. Total Nitrogen Hydroxide reagent( Hach Cat No. 26717-25)

4.2. Total Nitrogen Persulfate Reagent power pillows (Hach Cat.No. 26718-49)
4.3. Total Nitrogen Reagent A Power Pillows (Hach Cat.No. 26719-49)
4.4. Total Nitrogen Reagent B Power Pillows (Hach Cat.No. 26720-49)
4.5. Total Nitrogen C Vials (Hach Cat.No.26721-25)
5. PROCEDURE
5.1 Turn ON the COD Reactor. Heat to 103-106 ° C (Optimum temperature is

105 °C)
5.2 Take Two Nitrogen Hydroxide reagent vials.( one Blank and second
sample).Using a funnel, add one total Nitrogen Persulphate Reagent Power
Pillows to each of two Nitrogen Hydroxide Reagent Vials.
GLAB-72 (New) - Total Nitrogen (HACH method) Page 2 of 3
Page 273
5.3 Add 2.0 ml of sample to one vial and 2.0 ml of DM water to another vial. Cap
both vials and shake vigorously for 30 seconds.
5.4 Place the vials in the pre heated COD reactor. Heat of 30 minutes.
5.5 Remove the hot vials from the reactor and allow to cool to room temperature.
5.6 Power ON Hach Spectrophotometer and enter program # 350 rotate the
wavelength to 410 nm.
5.7 Remove the caps from the cooled digested vials and add the content of one TN
Reagent A power pillows to each vials. Cap vials and shake for 15 seconds.
Press shift + Timer. A three minutes reaction period will begins.
5.8 After the timer beeps, remove the caps from the cooled vials and add one TN
Reagent B power pillow to each vials. Caps vial and shake for 15 seconds.
Press Shift + Timer .A Two minutes reaction period will begins
5.9 After the timer beeps, Take two Total Nitrogen Reagent C vials, (one is blank
and other Sample) add 2.0 ml of digested treated sample to one vial. Add
2.0ml of treated reagent blank to the second Total Nitrogen C Vials.
5.10 Cap vials and slowly invert 10 times to mix. Use slow, deliberate inversions for
complete recover. The vials will be warm.
5.11 Press Shift + Timer .A five minute reaction period will begin. Do not invert the
vial again. When the timer beeps, place the COD vials adapter into the cell holder
with the marker to the right. Clean outside of the blank. Place the blank into the
adapter .Place the cover on the adapter. Press Zero.The display will show 0 mg/l TN
TNT. Place the sample vial into the adapter and cover the adapter. Press READ. The
display will show the result in mg/l NITROGEN will be displayed.
6. CALCULATION
Total Nitrogen, as mg/l Nitrogen = Reading displayed on the spectrophotometer.
GLAB-72 (New) - Total Nitrogen (HACH method) Page 3 of 3

Page 274
SECTION : WATER
TOTAL ORGANIC CARBON, TOTAL CARBON & INORGANIC CARBON

Page 275

TOTAL ORGANIC CARBON, TOTAL CARBON & INORGANIC CARBON
G-LAB METHOD No.73
(By K.P. Abraham)
1. SCOPE
This test method covers the determination of Total Carbon (TC), Inorganic Carbon (IC), Total
Organic Carbon (TOC), Non Purgeable Organic Carbon (NPOC), and Purgeable Organic
Carbon (POC) in water, drinking water, waste water, sea water, process water, make up water
etc. in the range from 4 µg/L to 4000 mg/L. This test method is applicable only to
carbonaceous matter in the sample that can be introduced into the reactive zone. The syringe
needle or injection opening size generally limit the maximum size of particles that can be
introduced. These measurements can be used for the monitoring of waste water treatment
process and for the monitoring of organic pollutants in Industrial waste water. The relationship
of TOC to other waste quality parameters such as Chemical Oxygen Demand (COD),
Biochemical Oxygen Demand (BOD), and Total Oxygen Demand (TOD) are much helpful in
studying the water chemistry.
2. PRINCIPLE
The filtered sample is homogenized and diluted as necessary and a micro portion is injected
into a combustion tube, which is filled with oxidation catalyst and heated to 680°C. The water
is vapourized and the organic and inorganic carbon in the sample are combusted or
decomposed to become CO2. This CO2 is transported by the high purity air (carrier gas)
through the IC reaction vessel and cooled and dried by a dehumidifier. It is then sent through a
halogen scrubber into a sample cell set in a Non Dispersive Infra Red (NDIR) gas analyzer,
where CO2 is detected. The NDIR output a detective signal (analog signal) which generates a
peak whose area is calculated by a data processor. The peak area is proportional to the TC
concentration of the sample.
Because total carbon is measured, IC must be measured separately and TOC obtained by
difference. Measurement of IC is done by Injecting the sample into the IC reaction vessel
containing phosphoric acid, where carrier gas flowing in as tiny bubbles. Only IC component in
the sample is decomposed to become CO2, which is carried to the NDIR, where it is measured.
Under these conditions organic carbon is not oxidized and only IC is measured. Carbon in the
form of carbonates and hydrogen carbonates are measured as IC.
TOC concentration can be obtained by subtracting the IC concentration from the TC

concentration. For the measurement of NPOC, acidify the sample and purge for a specified
time with high purity synthetic air and then do the TC measurement. This will give the NPOC
reading.
The difference between TOC and NPOC can be considered as POC.
GLAB-73 (Revised) - Total Organic Carbon, Total Carbon & Inorganic Carbon Page 2 of 5
Page 276
3. APPARATUS
3.1. Total Organic Carbon analyzer,

Shimadzu TOC - 5000
3.2. Syringes, 250 µ1 & 2500 µ1
3.3. Filtration apparatus
3.4. Filter paper, 0.45 µm pore size
3.7 volumetric flasks, 1 L
3.8 Drying oven
4. REAGENTS
4.1 Carbon free water

De mineralized water sparged with purified synthetic air.
4.2. IC reagent (Phosphoric acid 25%) Use ready made solution, part no. 630—00710.
Alternatively use analar grade phosphoric acid solution.
4.3. TC catalyst (Normal & High sensitivity) Normal - TC catalyst set P/N 638-92069-01
High sensitivity - H.S TC catalyst set P/N 638 – 92070 - 01
4.4. Hydrochloric acid, 2N Part No. 630 - 00998
Alternatively use 2N hydrochloric acid solution.
4.5. Organic carbon, stock solution (lml=lmg C)
Dissolve 2.1254 g of anhydrous potassium hydrogen (KHC8H4O4) in carbon free water
and dilute to 1 L.
4.6. Inorganic carbon, stock solution (lml=lmg C) Dissolve 4.4122 g anhydrous sodium
Carbonate (Na2CO3) 3.497g anhydrous sodium bicarbonate (NaHCO3) and dilute to 1 L.
4.7. Calibration standards (10 mg/L & 100 mg/L)
Prepare diluted standards of organic carbon and inorganic carbon from the respective
stock solutions using carbon free water.
4.8. Carrier gas
High purity synthetic air.
4.9. Purging gas
CO2 free synthetic air.
5. CALIBRATION
5.1 Supply carrier gas to TOC-500 by pressure 5-6 Kg/cm2G from a synthetic air gas
supply.
5.2. Turn ON the power switch on the right side of the instrument.
5.3. Depress NEXT key to go to MAIN MENU.
5.4. Confirm the pressure of carrier gas is 4-5 kg/cm2G and flow rate is 150ml/min.
Adjust them if necessary.
NOTE: Open the front door to adjust the carrier gas pressure controller and the
main flow controller.
5.5. Go to GENERAL CONDITIONS screen by either moving the curser to the item
number(#) or inputting the item directly and pressing enter.
5.6 Set TC FURNACE to ON. Check if other items have correct settings. If necessary,
conduct setting again. Then, call the MAIN MENU screen and move the cursor to #1
CALIBRATION and press enter to bring the CALIBRATION screen.
Page 277
5.7. Set the CALIBRATION conditions by entering the standard concentrations and other
parameters requested In the screen.
5.8. Depress NEXT key to go to MEASUREMENT START screen.
5.9. Put the first standard solution on the sample pan, insert the sampling tube with
sampling needle into it, and then depress START/STOP key. The standard solution is
automatically measured under the conditions set on CALIBRATION/CONDITIONS
screen. The progress of the measurement, a peak, and a peak area are then displayed.
When the sign of completion (COMPLETED) is displayed, depress NEXT key. Keep
the next standard and depress START/STOP to start the measurement of the second
standard.
When the measurement of all the standards specified in the SANDARD/CONDITIONS

are over, go to CALIBRATIN CURVE screen by pressing NEXT key.
5.10. Designate shifting of calibration curve to the zero point (SHIFT TO ORIGIN) and the
PROTECT processing to store calibration curve data (PROTECT), if necessary.
5.11. Go to MAIN MENU from where the sample measurement can be initiated.
6. PROCEDURE
6.1. Supply carrier gas to TOC-5000 with pressure of 5-6 Kg/cm2G from a synthetic air
gas supply.
6.2. Turn ON the power switch on the right side of the instrument.
6.3. Depress NEXT key to go to MAIN MENU.
6.4. Confirm the pressure of carrier gas is 4-5 Kg/cm2G and flow rate is 150 ml/min.
Adjust them if necessary.
NOTE: Open the front door to adjust the carrier gas pressure controller and the
main flow controller.
6.5. Go to GENERAL CONDITIONS screen by either moving the curser to the item
number(s) or inputting the item directly and pressing enter.
6.6 Set TC FURNACE to ON. Check if other items have correct settings. If necessary,
conduct setting again. Then, call the MAIN MENU screen and move the cursor to #2
SAMPLE MEASUREMENT and press enter.
6.7. Set the analysis conditions by entering the datas like, calibration curve to be used,
sample volume, dilution factor, No. of injections etc.
6.8. Depress NEXT key to go to MEASUREMENT START screen.
6.9. Put the sample on the sample pan and insert the sampling tube with sampling needle
into it, then depress START/STOP key. The progress of the measurement, a peak,
and a peak area are then displayed. When the analysis is over, sign of completion
(COMPLETED) is displayed.
6.10. If the measuring items or other samples to be measured remain, depress NEXT key.
When measuring items remain, the present screen starts again, and the measurement
for these items is performed. When measuring items do not remain, the screen returns
to SAMPLE MEASUREMENT/CONDITIONS and the number of the next sample is
displayed. So, proceed to the measurement of the next sample.
6.11. When measurement of every sample has been completed, depress END key to go to
#5 DATA REPORT screen via MAIN MENU.
6.12. As all the result of every measurement of sample is displayed print it out by
depressing PRINT key.
6.13. When operation of the equipment is to be terminated go to #7 STANDBY OPTIONS
screen via MAIN MENU screen.
6.14. To stop the equipment, depress 1 and press ENTER, and then press STANDBY key.
Page 278
NOTE: After measuring sample which contains large amount (about 1,000 ppm or
more) of acid or salt, if equipment is left with sample remaining in sample
injection needle, the needle may be corroded by sample or clogged with
precipitated salt. To prevent this, execute TC and IC WASHING with
sampling tube end put in DM water, using MECHANICAL CHECK
function on MAINTENANCE screen.
6.15. When, the indication of “The remaining time count before the power switch may be
turned off” becomes 0 (zero), turn off the power switch and the carrier gas supply.
7. CALCULATIONS
7.1. Total Carbon (TC)

As displayed and printed after measurement by the instrument when the sample is
directly analysed in the TC mode.
7.2 Inorganic Carbon (IC)

As displayed and printed after measurement by the instrument when the sample is
directly analysed in the IC mode.
7.3 Total Organic Carbon (TOC)

Instrument calculate and prints the result in the report after the analysis.
7.4 Non Purgeable Organic Carbon (NPOC)

As displayed and printed after measurement by the instrument when the acidified
and purged sample is analysed in the NPOC mode.
7.5 Purgeable Organic Carbon (POC)

POC = TOC-NPOC
7.6 Dissolved CO2

Difference between the IC, when the sample is analysed before purging and after
purging in the IC mode.
7.7 Dissolved Carbon (DC) and Dissolved Organic Carbon (DOC)

When the analysis is carried out in filtered samples, the TC and TOC will
represent DC & DOC respectively.
Reference : ASTM D2579

APHA 5310 B
Page 279
G-STATIONLABORATORY METHOD
SECTION : WATER
THE DETERMINATION OF TURBIDITY
(NEPHELOMETRIC METHOD)

Page 280

TURBIDITY (NEPHELOMETRIC METHOD)
G-LAB METHOD No.74
(By K.P. Abraham)
1. SCOPE
This method covers the determination of relative turbidity of cooling water, industrial
wastewater and sea water. This method can be used for low turbidity measurement. Clarity of
water is important in producing products destined for human consumption and many
manufacturing uses. The turbidity of water is a major determinant of the condition of water.
2. PRINCIPLE
This method is based on a comparison of the intensity of light scattered by the sample under
defined conditions with the intensity of light scattered by a standard reference suspension
under the same conditions. The higher the intensity of the scattered light, the higher the
turbidity. Formazin polymer is used as the reference turbidity standard suspension.
3. APPARATUS
3.1. Turbidimeter
Turbidimeter by HACH Model 2100 A
3.2. Sample cells, 25 mm
3.3. Gelex secondary standards
3.4. Filtering apparatus
3.5. Pipettes.
4. REAGENTS
4.1. Formazin standard, 4000-NTU (stock solution) As supplied by the Nephelometer

supplier.
4.2. Formazin standard, 1000 NTU

Pipette 25 ml of well-mixed formazin stock solution into a clean 100 ml volumetric
flask and dilute to the mark with turbidity free water and mix well. Turbidity free
dilution water can be prepared by filtering distilled water through 0.45 micron
membrane filter.

Pipette 2.5 ml of the stock solution to a clean 100 ml volumetric flask and dilute to the
mark with turbidity free water and mix well.

Pipette 0.25 ml of the well-mixed stock solution into a clean 100 ml volumetric flask
and dilute to the mark with turbidity free water and mix well.
4.5. Formazin standard, 1.0 NTU

Pipette 1.0 ml of well mixed 100 NTU standard prepared in Step 4.3. into another
clean, class A 100 ml volumetric flask and dilute to the mark with turbidity free water.
Mix well. This is a nominal 1.0 - NTU standard.
GLAB-74 (Revised) - Turbidity (Nephelometric method) Page 2 of 5

Page 281
4.6 Using the 0 - 1.0 NTU secondary standard, standardize the instrument on the
1.0 NTU range and then measure the turbidity of the dilution water.
4.7. Using the following equation, calculate the actual turbidity of the nominal 1.0 NTU
standard prepared in step 4.5.
T = (A/B) (Ts) + (1 - A/B) (Tw)
Where, A = volume of stock solution

B = total diluted volume
Ts = turbidity of standard
Tw = turbidity of dilution water
For example with dilution water of 0.15 NTU:

T = (1/100) (100) + (1 - 1/100) (0.15)
T = 1 + 0.1485
T = 1.15 NTU
5. CALIBRATION
5.1. Primary calibration
5.1.1. Switch ‘ON’ the instrument and allow the instrument to warm up for 30
minutes.
5.1.2. Place the cell riser in the cell holder and insert the 0 - 1000 NTU
standard. Standardize the turbidimeter to give a reading equal to the value
of the standard.
5.1.3. Place the 0 - 100 NTU standard in the cell holder and select the range. A
reading within 2 NTUs of the standard value should be obtained. Range
adjustment is required if the reading is in error.
5.1.4. Remove the cell riser and repeat the procedure with 0 - 10 and 0 - 1.0
NTU standards. Determine if range adjustment is necessary.
5.2 Range adjustment
5.2.1. If the instrument has been on for several days proceed to Step
5.1,5.3.If not proceed as follows: Turn the instrument on and adjust to
range 0.2. Fill a sample cell with tap water, place it into the cell holder
and cover with the light shield. Allow the instrument to stand for at least
12 hours.
5.2.2. Remove the sample cell.
5.2.3. Turn the range select knob to 1000, place the cell riser in the cell holder,
insert the 0 - 1000 NTU standard and cover with the light shield.
5.2.4. Turn the STANDARDIZE control on the front panel fully counter
clockwise and note the meter reading. Then turn the control clockwise
until twice this reading is obtained. Leave the STANDARDIZE control in
the position for calibrations of all ranges except the 0 - 0.2 range.
5.2.5. Adjust the 1000 range trimmer potentiometer to obtain a meter reading
equal to the NTU value of the 0 - 1000 standard. To do this, open the
base panel and adjust the 1000 potentiometer. (The 1000 NTU range
must always be adjusted first.)
5.2.6. With the cell riser still in place, insert the 0-100 NTU standard, cover
with the light shield and set the range selector to 100. Adjust the 100
NTU range trimmer potentiometer to obtain a reading equal to the NTU
value of the 0-100 standard.

Page 282
5.2.7. Remove the cell riser and insert the 0 - 10 NTU standard, cover with the
light shield and switch the range selector to 10. Adjust the 10 NTU range
trimmer potentiometer to obtain a meter reading equal to the NTU value
of the 0-10 standard.
5.2.3. With the cell riser removed from the instrument insert the 0 - 1.0 NTU
turbidity standard, cover with the light shield and set the range selection
switch to 1.0. Adjust the 1.0 NTU range trimmer potentiometer to obtain
a reading equal to the NTU value of the 0 - 1 standard.
5.2.9. Use water containing less than 1.0 NTU turbidity. Tap water may be
adequate. With the cell riser removed from the instrument, place a sample
tube of this water in the cell holder. Cover with the light shield and adjust
the STANDARDIZE control to obtain a reading of 0.20 NTU on the
1.0NTU range. (This may not be possible if the water is too turbid to
reach this value. In this case, it will be necessary to dilute the turbid
sample with very clear water in a proportion so that a reading of 0.20 can
be obtained by adjusting the STANDARDIZE control.) With the
STANDARDIZE control adjusted to give a reading of 0.20 on the 1.0
range, turn the range selector to 0.2. Adjust the 0.2 trimmer potentiometer
to obtain an exact full-scale meter reading. This completes the range
adjustment. A recheck may be made of the adjustments.
5.2.10 Restandardize the instrument, using the appropriate turbidity standard.
5.3. Calibrating secondary standards
5.3.1. When primary calibration adjustments are complete, the Gelex secondary
standards should be measured to establish a true turbidity value for the
secondary standards on this instrument. This will provide a calibrated
value traceable to the formazin primary standards.
5.3.2. Place the secondary standards in the instrument with their index mark
aligning with the mark on the spill ring and select the corresponding
ranges marked on the vials. Read the true values displayed and mark
them on the vials.
6. PROCEDURE
6.1. Standardization should be performed before each series of measurements to assure

accurate results. Insert the secondary standard for that range with index mark to the
front (use cell riser when using the 100 NTU and 1000 NTU range standards), cover
with the light shield and adjust the STANDARDIZE control until the meter reading
equals the value of the standard. If you change measurement ranges, the new range
also should be standardized before measurements are taken.
6.2. Fill a clean sample cell to the mark with well-mixed sample and place t into the cell
holder. The sample cell must be clean, dry and free of fingerprints.
6.3. Insert the sample in the instrument, aligning the cell index mark with the raised
mark on the spill ring around the cell holder opening. Be sure the cell is down
completely and held in place the spring clip. Cover the sample with the light shield.
6.4. Use cell riser when using the 100 NTU and 1000 NTU ranges.
6.5. Select an appropriate range. If the approximate sample turbidity is unknown, begin
with the highest range and work down. Wait at least 15 seconds in each range to
allow the instrument to stabilize. Select the lowest range possible without having an
overange condition.
Page 283
6.6. Read the turbidity of the sample from the digital display.
6.7. It may be necessary to dilute the sample in order to bring it within the range of the
instrument when measuring highly turbid samples. When dilution is necessary, the
sample should be diluted with another portion of sample that has been filtered
through 0.45 micron membrane filter.
7. CALCULATION
7.1. For undiluted sample

Nephelometric turbidity units (NTU) = Instrument reading
7.2. For diluted samples
Nephelometric turbidity units (NTU) = A x (B+C)
C
Where
A = NTU found in diluted sample
B = Volume of dilution water ml
C = Sample volume taken for dilution ml

APHA 2130 B
Manual for Turbidimeter.

JEBEL ALI POWER STATION
G STATION LABORATORY
P.O.BOX 564, DUBAI
UNITED ARAB EMIRATES
Tel: 00971-4-8044888, Fax: 00971-4-8846482

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DUBAI ELECTRICITY AND WATER AUTHORITY

G - STATION LABORATORY METHODS

GLAB - 01 Alkalinity, Caustic Page 1

GLAB - 28 Corrosion rate (Coupon method) Page 104

GLAB - 56 Quality checks, acidity & alkalinity by autotitrator Page 2 15

DUBAI ELECTRICITY AND WATER AUTHORITY

G-STATION LABORATORY METHOD

THE DETERMINATION OF CAUSTIC ALKALINITY

Method No. : GLAB - 01

DUBAI ELECTRICITY & WATER AUTHORITY Laboratory Methods of Analysis

TEST METHOD FOR WATER

4.1. Barium chloride solution (10%)

GLAB-01(Revised) - Caustic Alkalinity Page 2 of 3

DUBAI ELECTRICITY & WATER AUTHORITY Laboratory Methods of Analysis

0.01M Sulphuric acid = 0.02N Sulphuric acid

Caustic alkalinity as CaCO3, mg/l = Titre reading x 0.02 x 50 x 1000

Caustic alkalinity as NaOH, mg/l = Titre reading x 0.02 x 40 x 1000

GLAB-01(Revised) - Caustic Alkalinity Page 3 of 3

DUBAT ELECTRICITY AND WATER AUTHORITY

G-STATION LABORATORY METHODS

DUBAI ELECTRICITY & WATER AUTHORITY Laboratory Methods of Analysis

TEST METHOD FOR WATER

3.1 Automatic micro burette, 10 ml

GLAB-02 (Revised) - Alkalinity Page 2 of 4

DUBAI ELECTRICITY & WATER AUTHORITY Laboratory Methods of Analysis

GLAB-02 (Revised) - Alkalinity Page 3 of 4

DUBAI ELECTRICITY & WATER AUTHORITY Laboratory Methods of Analysis

Result of Hydroxide Carbonate Bicarbonate

P = Phenolphthalein alkalinity, M = Methyl orange alkalinity.

For drinking water P < ½M.

Reference APHA 2320 B

GLAB-02 (Revised) - Alkalinity Page 4 of 4

DUBAT ELECTRICITY AND WATER AUTHORITY

G-STATION LABORATORY METHODS

DUBAI ELECTRICITY & WATER AUTHORITY Laboratory Methods of Analysis

TEST METHOD FOR WATER

A sample aliquot is Nesslerized directly and the ammonia content determined

3.2. Weighing balance

3.3. Measuring cylinder, 50 ml

3.5. Volumetric flask, 50 ml (10 Nos.), 1 lit, 200 ml.

3.6. Spectrophotometer, Jasco Model V 530

3.7. Beakers, 200 ml (10 Nos.)

3.8. Cell, 50 mm.

4.1. Nessler’s reagent.

Readymade solution as purchased or this reagent can be prepared as follows:

4.2. Ammonia solution, Standard (1 ml = 1 mg NH3)

DUBAI ELECTRICITY & WATER AUTHORITY Laboratory Methods of Analysis

NH4C1 and dissolve in distilled water and make up to 11.

Pipette 5.0 ml of the ammonia solution (1 ml = 1 mg NH3) to a 200 ml volumetric

Ammonia, mg/l as NH3 = Absorbance x F

c) As the intensity of colour fades with time, it is important to take

Reference: ASTM D 1426

GLAB-03 (Revised) - Ammonia Page 3 of 3

DUBAI ELECTRICITY AND WATER AUTHORITY

G-STATION LABORATORY METHOD

THE DETERMINATION OF AMMONIA

Method No. : GLAB - 04

DUBAI ELECTRICITY & WATER AUTHORITY Laboratory Methods of Analysis