SOP'S FOR WATER

Sr. No. SOP's Name

1 OIL AND GREASE
2 BIO-ASSAY
3 ALKALINITY
4 ACIDITY
5 TOTAL HARDNESS
6 CALCIUM HARDNESS
7 PESTICIDES
8 SOLIDS
9 TOTAL SUSPENDED SOLIDS
10 TOTAL SOLIDS (TS)
11 VOLATILE SOLIDS & FIXED DISSOLVED SOLIDS
12 TOTAL DISSOLVED SOLIDS (TDS)
13 SULFATE
14 NITRATE
15 CHLORIDES
16 COLOUR
17 CONDUCTIVITY
18 CYANIDE
19 SALINITY
20 NITRITE NITROGEN
21 PHOSPHATE
22 FLUORIDE
23 PHENOL
24 DETERGENT IN WATER
25 HEXAVALENT CHROMIUM
26 SODIUM(NA)/POTASSIUM(k)

27 DETERMINATION OF MERCURY BY COLD VAPOUR ATOMIC ABSORPTION SPECTROMETRIC METHOD USING FIMS

28 DETERMINATION OF METALS BY AAS (ATOMIC ABSORPTION SPECTROMETER) OR ICP (INDUCTIVELY COUPLED PLASMA METHOD)

29 BIOCHEMICAL OXYGEN DEMAND (B.O.D.)

30 TOTAL RESIDUAL CHLORINE (T.R.C.)
31 SILICA
32 TOTAL KJELDAHL NITROGEN (T.K.N.)
33 AMMONIA
34 CHEMICAL OXYGEN DEMAND (C.O.D.)
35 BACTERIOLOGY
36 SULPHIDE
 WI No.
OIL & GREASE Rev. No. 0
Date 10.12.2013

Method : Extraction Method

Principle : N/A
Reagents: 1) 1 : 1 H2SO4
2) Hexane

Apparatus : N/A

Absorbing Media: N/A

Interferance : N/A

Sr. No. PROCEDURE RESPONSIBILITY

Take 500ml of sample is taken and acidified by adding 2-5 ml. 1 : 1 H2SO4.
1
To this add 25 ml Hexane in separating funnel.

Shake vigorously for few minutes, then allow to stand. Organic and aqueous
2
phase is separated.

Organic solvent is collected in preweighted evaporating dish. With remaining

3
aqueous phase; the same procedure is followed.

4 If you find emulsion, it is broken down by adding Isopropanol or Acetone. JSO/JSA

Finally the Organic extract containing the oil fraction is filtered through sodium
5
sulphate via funnel into preweighed evaporating dish.

Keep on water bath so that Hexane is evaporated and keep in oven for 10
6
minutes.

7 Take the final weight of Dish (W2).

Final wt. Of dish (W2) – Initial of (W1) X 1000

Calculation :
ml of Sample

Reference: Water APHA 19th Edition

Date Of Change Description of Change Prepared By Reviewed By Approved By

 WI No.
BIO-ASSAY Rev. No. 0
Date 10.12.2013
Static Bio-assay Method For relatively stable substances when dissolved oxygen (DO) in the Diluent is
insufficient or when surface oxygen absorption does not maintain a DO level adequate for the test
Method : organisms oxygen may be supplied by initial oxygenation of the Diluent controlled artificial oxygenation of
test solutions or periodic renewal of test solutions

Bioassay are conducted to evaluate the toxicity of effluents or other materials In a Bioassay experimental
fishes are subjected to a series of concentrations of a known or suspected toxicant under adequately
controlled conditions. Measures of toxicity
Principle : Bioassay results using fish shall be expressed in terms of tolerance limits (II) TL 50 and TL90 . e g 96hrs TL50
of a toxic sample is that concentration in which 50% of the fish survive for 96hrs TL50 is the standard
measure of toxicity and is equivalent to the median tolerance limit

Guppies Experimental fish should be those species adaptable to laboratory conditions and those of which
adequate numbers of a usable size can be maintained. Specimens of 1-2 inches in length are preferable.
Type Of Fish : The length of the largest individual should be no more than 1 5 times, the length of the smallest.

Fish stock may be kept in water of a quantity and quality such that they will remain in good condition Fish
selected for testing should be acclimatized to the test temperature and the experimental dilution water for
at least 10 preferably 30 days. Adequate aeration of the water is essential The fish should be fed daily or at
least 3 times per week regularly, but they should not be fed for 2 days before testing Incidence of death or
disease among acclimatized fish within a period of 4 days immediately preceding the test must be less than
Preparation of Test 5%.
Fish :
Dilution Water: A carbon-filtered tap water, which is acclimatized, may be used Synthetic dilution waters of
a constant and reproducible quality capable of supporting aquatic life may be prepared by the addition of
appropriate chemicals to distilled water Samples should be stored in polythene Carboys at 4'C.

Absorbing Media:
Interferance :
Sr. No. PROCEDURE RESPONSIBILITY

Temperature : The range of temperature must not exceed 4 ̊C. 25 ± 2 ̊C is

1
recommended.

2 Dissolved : Must not fall below 4 mg/Liter Oxygen

Content pH : Should be between 6.5 to 7.5 Determination pH, Temperature and

3 DO should be done before the experiment and after every 24 after experiment.

Sample Dilution : 100%, 56%, 32%, 18%, 10%, 1%, 0.56%, 0.32%, 0.18% etc.
4
Dilutions are made with acclimatized water.

Controls : A concurrent control test must be performed in exactly the same

manner as the sample tests and under the same conditions. Diluent alone is
5 used as the medium in which control fishes are held. There must be no more
than 10% mortality during the test and at least 90% must remain apparently
healthy.
JSO/JSA
Number of Test Fish : At least 10 fish will be used. Fish should not be fed
during test period of 96 hours. Temperature, pH, Dissolved oxygen and the
6
number of dead fish in each test container must be observed and recorded at the
end of each 24 hours period after introduction of the fish

Test Containers : Should be of glass, Average depth of the liquid in the test
7
container must be uniform in all parallel tests and must not be less than 15cm.

Total Fish Weight and Liquid Volume : The weight of all fish in a test container
8
must not exceed 1 gm per liter.

Results : Plot a graph of the data on semi logarithmic coordinate paper with
concentrations on the logarithmic and % survival on the arithmetic axis. A straight
line is drawn between 2 points representing survival at the two successive
9
concentrations that were lethal to more than half and to less than half of the fish.
The conc. at which this line crosses the 50% survival line is the TL50 value.

Calculation : N/A

Reference: N/A

Date Of Change Description of Change Prepared By Reviewed By Approved By

 WI No.
ALKALINITY Rev. No. 0
Date 10.12.2013
Method : Titrimetric method

Principle : N/A

Reagents: 1) 0.1N Sulphuric Acid :- Take 2.8ml concentrated H2SO4 dilute to 1000 ml with distilled water. 2)
0.02 N H2SO4 Solution :- 200ml of 1N Sulphuric Acid diluted to 1000ml
3) Methyl orange indicator

Apparatus : N/A

Absorbing Media: N/A

Standardization : 1 .060gm OF sodium Carbonate diluted to 1000ml with distilled water, 10ml of above solution titrate with
0.02N H2SO4. End point to orange.

Sr. No. PROCEDURE RESPONSIBILITY

Take 50ml sample + 0.1 ml or 2 drops of methyl orange indicator => Titrate with
1 0.02 N H2SO4. End point is yellow to orange. JSO/JSA

Total Alkalinity as CaCO3= [B.R x 0.02 x 50000 ]

Calculation : ml of Sample

Reference: Water APHA 19th Edition

Date Of Change Description of Change Prepared By Reviewed By Approved By

 WI No.
ACIDITY Rev. No. 0
Date 10.12.2013
Acidity of water is its quantitative capacity to react with a strong base to a designated pH and strong
DISCUSSION : mineral acids, weak acid such as carbonic, acetic and hydrolyzing salts. Such as iron or aluminum sulfates
may contribute to the measured acidity according to the method of determination.

Hydrogen ions present in a sample as a result of dissociation or hydrolysis of solutes react with additions of
Principle : standard alkali.

Significance : Acids contribute to corrosiveness and influence chemical reaction rates. Chemical speciation and biological
process. The measurement also reflects a change in the quality of the source water.
Method : Titrimetric method

Apparatus : N/A
1) 0.02 N NaOH Solution :- 0.8 gm of NaOH pallets to 1000ml distilled water.
Reagents : 2) Phenolphthalein Indicator.

Absorbing Media: N/A

Interferance : N/A
Standardization : N/A

Sr. No. PROCEDURE RESPONSIBILITY

Take 50ml sample + 2 to 3 drops of phenolphthalein indicator.

1

2 Titrate against 0.02 N NaOH solution. JSO/JSA

3 End point is colourless to pink.

Calculation : Acidity as mg/l CaCO3 = A X N X 50000 = B.R x 20

ml sample
A = Burette reading
B = Normality of NaOH

Reference :
Water APHA 19th Edition

Date Of Change Description of Change Prepared By Reviewed By Approved By

 WI No.
TOTAL HARDNESS Rev. No. 0
Date 10.12.2013
Method : Titrimetric method

EDTA form chelated soluble complex when added to a solution of Certain metal cations. If small amount
of Eriochrome Balck is added to an aqueous solution containing Ca and Mg ions at pH 10 + 0.1, the
Principle : solution becomes wine red. After titrating with EDTA Ca and Mg will be complex the solution turns from
wine red to blue.

1) Buffer Solution :- Dissolve 1.179 EDTA and 780mg of Magnesium Sulphate ( MgSO4.7H2O) in 50ml
distilled water, Add this solution to 16.9gm NH4Cl and 143ml conc. NH4Cl with mixing and dilute to 250 ml
with distilled water.
Reagents: 2) 0.01 M
EDTA Solution :- 3.723 gm of EDTA in 1000ml Distilled Water.
3) Eriochrome Black T :-0.5
gm of dye i.e. Eriochrome Black T + 100 gm of Sodium chloride => grind to fine powder . This will serve as
an indicator.

1 gm of anhydrous Calcium Carbonate in 500ml beaker + 1:1 HCL until CaCO3 dissolves + 200ml distilled
Standardisation : water, boil for few minutes, cool and add indicator + 3N NH4OH to adjust orange colour => dilute to 1 lit

Apparatus : N/A

Absorbing Media: N/A

Interferance : N/A

Sr. No. PROCEDURE RESPONSIBILITY

Take 50ml sample + 1 ml buffer solution to give pH 10.0 +
A pinch of Eriochrome Black T indicator. Titrate with 0.01M EDTA. End point is
1 wine red to blue JSO/JSA

A x B x 1000
Hardness as CaCO3mg/l = -------------------------------
Calculation : ml of sample

A= Burette Reading B = mg of CaCO 3 equivalent to 1ml of EDTA

Reference: Water APHA 19th Edition

Date Of Change Description of Change Prepared By Reviewed By Approved By

 WI No.
CALCIUM HARDNESS Rev. No. 0
Date 10.12.2013
Method : Titrimetric method

Principle : N/A
1) 1 N NaOH :- 40gm of NaOH pallets dissolve in 250ml watr and diluted to 1000ml with distilled water.
2) 0.01 M EDTA
Solution :- 3.723 gm of EDTA in 1000ml Distilled Water.
3) Muroxide INdicator :- 200mg of
Reagents: muroxide indicator + 100gm Sodium Chloride => grind to fine powder.

Standardisation : N/A

Apparatus : N/A

Absorbing Media: N/A

Interferance : Calcium Hardness N/A

Sr. No. PROCEDURE RESPONSIBILITY

Take 50ml sample + 2 ml NaOH(1N) to produce pH 12 to 13. Stirr and add pinch
1 of Muroxide indicator. Titrate with 0.01M EDTA. End point is pink to violet. JSO/JSA

Ca as CaCO3 md/l = A x B x 1000

Ax
B x 1000 ml of sample

Hardness as
Calculation : CaCO3mg/l =
A= Burette Reading
------------------------------
B = mg of CaCO 3 equivalent to 1ml of EDTA
Magnesium- Hardness
Calculation for Ca ions = A x B x 400.8
ml of sample
ml of sample
A= Burette Reading B = mg of CaCO 3 equivalent to 1ml of EDTA

Reference: Water APHA 19th Edition

Date Of Change Description of Change Prepared By Reviewed By Approved By

 WI No.
PESTICIDES Rev. No. 0
Date 10.12.2013
The pesticides are extracted with a mixed solvent, Methylene chloride/hexane.
Method : The extract is concentrated by evaporation.
The individual pesticides then are determined by gas chromatography.

Principle : N/A
Apparatus : N/A

1) Hexane
2) Ethyl acetate
3) Methylene Chloride
4) Sodium Sulphate anhydrous
5) Nitrogen gas, purified grade, moisture and oxygen free
6) Pesticide Reference Standards
Reagents: a) Stock Pesticide Solution : Ready made stock solution.

b) Working Standard Solution : Prepare final concentration of standards in hexane solution as

required by detector sensitivity and linearity.

Absorbing Media: N/A

Interface : N/A

Standardization N/A

Sr. No. PROCEDURE RESPONSIBILITY

Pour 1 Lt of sample into 2 Lt separating funnel after adjusting the pH to 6.5 – 7.5
1 with sulfuric acid or sodium hydroxide as per the requirements.

2 Extract the sample thrice with 60ml of 15% methylene chloride in hexane.

JSO/JSA
Filter the organic phases through anhydrous sodium sulphate into dry conical
3 flask. Evaporate to dryness.

Add 2 ml of n-hexane of GC grade. Swirl and analyze the extract by G.C.

4

mg/l = Conc. in mg from G.C. Peak x 2 x 1000

Calculation : '106

Reference : APHA 19th Edition

Date Of Change Description of Change Prepared By Reviewed By Approved By

 WI No.
PESTICIDES Rev. No. 0
Date 10.12.2013
Analysis of suspended solids estimates the total amount of particulate matter in a sample. Solids refer to
matter suspended or dissolved in water or wastewater. Solids may affect water or effluent quality
adversely in a number of ways. Waters with high generally are of inferior palability & may induced an
unfavorable physiological react_i_Titeisral.e.. transient consumer. For these reasons, a limit of 500mg
Introduction :
dissolved solids/lit is destra e for drinking waters. Solids analysis are important in control of biological &
physical wastewater treatment processes & for assessing compliance with regulatory agency wastewater
effluent limitations.

Sampling : Garb sampling

Use resistant glass or plastic bottles provided that the material in suspension does not adhere to container
Handeling and walls. Refrigerate sample at 4°C upto analysis to minimize microbiological decomposition of solids.
Preservation :

Absorbing Media: N/A

Sr. No. PROCEDURE RESPONSIBILITY

1 N/A JSO/JSA

Calculation : N/A

Reference : APHA 19th Edition

Date Of Change Description of Change Prepared By Reviewed By Approved By

 TOTAL SUSPENDED WI No.
Rev. No. 0
SOLIDS Date 10.12.2013

Method : Gravimetric method

A well mixed sample is filtered through a weighed standard glass fiber filter paper & residue retain on the
Principle : filter paper is dried to a constant weight at 103 to 105°C. The increase in weight of the filter paper
represents the TSS.

Glassware : Beakers, Measuring Cylinder

Reagents:
Glass Fiber Filter paper 47 mm.

Instruments/
Equipments : Drying Oven, Desiccators, Analytical balance & Filtration assembly

Removal of Exclude large flotation particles or submerged agglomerates of non-homogeneous materials from the
Interferences : sample For samples of high dissolved solids thoroughly wash the filter to removal of the dissolved
materials.

Standardization N/A

Sr. No. PROCEDURE RESPONSIBILITY

Turn on the balance fifteen minutes prior to weighing of filter papers by
1 depressing the bar. Place the date of the analysis under "Date Initial- on the data
sheet

Set balance to zero by depressing the re zero bar.Weigh the filter paper on the
2
balance to four decimal places

Record the result on the data sheet under "Initial Weight". Place & label the filter
3
paper. Now place the filter paper on the Filtration assembly

Pour the well mixed & measured sample on the paper. Wait sometime so as to
4
drain out the water sample completely. JSO/JSA

Remove the filter paper & keep it in the oven. Care should be taken to see that
5
paper should not be scratched. (Dry up to constant weight)

After drying take filter paper from oven and Allow to cool to room temperature in
6
desiccator

It is essential that the filter paper is in a horizontal position during this transfer.
Particulate matter is not stable on the membrane surface and can be dislodged.
7 Record weight on data sheet under "Final Weight" next to the corresponding
initial weight. Record the sample the 'Sample id' column.

(W2 - W1) x 1000 x 1000

Calculation : ml of Sample (V)
W2 : Final wt. of filter paper in grams W1 : Initial wt. of filter paper in grams V : Volume of sample

Reference : APHA 19th Edition

Date Of Change Description of Change Prepared By Reviewed By Approved By

 WI No.
TOTAL SOLIDS(TS) Rev. No. 0
Date 10.12.2013

Method : Gravimetric method

A well mixed sample is evaporated to dryness in a pre - weighed dish and dried to a constant weight at 103
Principle : to 105°C. The increase in dish weight represents the total solids

Glassware : Beakers, Measuring Cylinder, Porcelain dish

Reagents:
N/A

Instruments/
Equipments : Drying Oven, Desiccators, Analytical balance , Water Bath

Removal of Highly mineralized water with a considerable calcium, magnesium, chloride, sulfate content may be
Interferences : hygroscopic & require prolonged drying, proper desiccation & rapid weighing. Exclude large flotation
particles or submerged agglomerates of non-homogeneous materials from the sample. Disperse visible
floating oil & grease with a blender before withdrawing a sample for analysis.

Standardization N/A

Sr. No. PROCEDURE RESPONSIBILITY

Dry the clean dish in oven to 103 to 105 ̊C for 1 hour & keep in desiccator for
1
coolin) )to the room temperature & weigh before use.

Transfer a measured volume of well mixed sample to pre-weighed dish &

2
evaporate upto dryness on water bath.
JSO/JSA
Keep evaporated sample for lhour in an oven at 103 to 105°C, cool dish in
3
desiccator upto the room temperature.

Repeat drying, cooling, desiccating & weighing until a constant weight is

4
obtained.

(A - B) x 1000 x 1000
Calculation : ml of Sample (V)
A : Final wt. of filter paper in grams B : Initial wt. of filter paper in grams V : Volume of sample

Reference : APHA 19th Edition

Date Of Change Description of Change Prepared By Reviewed By Approved By

 VOLATILE SOLIDS & WI No.

FIXED DISSOLVED Rev. No. 0

SOLIDS Date 10.12.2013

Method : Gravimetric method

The residue of total solids is ignited to constant weight at 550 ̊C. The remaining solids represet the fixed
Principle : dissolved solids while the weight loss on ignition is the volatile solids.

Glassware : Beakers, Measuring Cylinder, Porcelain dish

Reagents:
Glass Fibre Filter paper 47 mm

Instruments/
Equipments : Drying Oven, Desiccators, Analytical balance , Water Bath and Muffle Furnace

Negative errors in volatile solids may be produced by loss of volatile matter while drying Determination of
Removal of low concentration of volatile solids in presence of high fixed solid concentration may be subject to
Interferences : considerable error.

Standardization : N/A

Sr. No. PROCEDURE RESPONSIBILITY

Take clean porcelain dish which is ignited in furnace. It is cooled in a desiccator

1
up to room temperature; take the initial weight (W1)

Take a known volume of sample filter through filter paper & take filtrate in an
2 empty porcelain dish which is already weighed. Evaporate this sample at 103 to
105°C until dryness.
JSO/JSA
Cool it in a desiccator & record the weight (W2). Again ignite this dish for 15 to 20
3 minutes in a muffle furnace whose temperature is maintained to 550°C until the
constant weight is attained.

Cool it in desiccator & record final weight (W3).

4

Volatile solids(mg/lit): (W2 - W3) x 1000 x 1000

V

Calculation : Fixed dissolved solids (mg/lit) : (W3 - W1) x 1000 x 1000

V
W3 :Wt. of ignited dish in grams W2 : Wt of residual dish in grams W1 : Wt. of empty dish

Reference : APHA 19th Edition

Date Of Change Description of Change Prepared By Reviewed By Approved By

 TOTAL DISSOLVED WI No.
Rev. No. 0
SOLIDS(TDS) Date 10.12.2013

Method : Gravimetric method

A well mixed sample is through a standard glass fiber filter paper, & the filterate is evaporated to dryness in
Principle : weighed dish and dried to constant weight at 180 ̊C . The increase in dish weight represents the total solids

Glassware : Beakers, Measuring Cylinder

Reagents:
Glass Fibre filter paper 47mm.

Instruments/
Equipments : Drying Oven, Desiccators, Analytical balance , Filtration assembly

Removal of Highly mineralized water with a considerable calcium, magnesium, chloride, sulfate content may be
Interferences : hygroscopic & require prolonged drying, proper desiccation & rapid weighing. Samples high in bicarbonate
require careful & possibly prolonged drying at 180 ̊C to insure complete conversion of bicarbonate to
carbonate.

Standardization N/A

Sr. No. PROCEDURE RESPONSIBILITY

Dry the clean dish in oven to 180 ̊C for 1 hour & keep in desiccator for cooling to
1
the room temperature & weigh before use.

Filter measured volume of welll mixed sample through filter paper, wash with
2
distilled water.
JSO/JSA
Transfer filterate to weighed evaporating dish & evaporate to dryness on water
3
bath .Dry for 1 hou in an oven at 180 ̊C, cool in desiccator & weigh.

Repeat drying, cooling, desiccating & weighing until a constant weight is

4
obtained.

(A - B) x 1000 x 1000
Calculation : ml of Sample (V)
A : Final wt. of filter paper in grams B : Initial wt. of filter paper in grams V : Volume of sample

Reference : APHA 19th Edition

Date Of Change Description of Change Prepared By Reviewed By Approved By

 WI No.
SULFATE Rev. No. 0
Date 10.12.2013
Method : Turbidimetric Method

Principle : Sulfate ion is precipitated in a hydrochloric acid medium with barium chloride to form barium sulfate crystals
of uniform size.

Apparatus : N/A

1) Conditioning Reagent : Mix 50 ml glycerol with solution Containing 30ml conc. HCl, 300ml distilled
water, 100 ml 95% ethyl or isopropyl alcohol and 75gm NaCl (Sodium Chloride).
2) Barium Chloride (BaCl2) : Crystals 20-30 mesh.
Reagents:
3) Standard Sulphate Solution : Dissolve 0.1479 g Sodium Sulphate Na2SO4 in d/w and dilute it to
1000ml. This solution is 100 ppm.
4) From the above solution prepare standards from 5ppm,10,15,20,25,30,35,40 ppm. Treat them same as
sample and read at 420nm.
Absorbing Media: N/A

Interference : Silica more than 500 ppm will interfere. Colour and Suspended matter in large amount will interfere.

Standardization N/A

Sr. No. PROCEDURE RESPONSIBILITY

Measure 100ml sample, (take dil. If necessary), add 5ml of conditioning reagent.
1

Add a spoonful of BaCl2, crystals, immediately stir for 1 min., constant speed, JSO/JSA

2 reading on spectrometer at wavelength 420 nm (charcoal treatment for coloured

samples) and filteration for turbid samples.

Calculation :- N/A

Reference :- APHA 19th Edition

Date Of Change Description of Change Prepared By Reviewed By Approved By

 WI No.
NITRATE Rev. No. 0
Date 10.12.2013
Method : Ultra Violate Spectrophotometer Screening Method

Principle : N/A

Apparatus : N/A

1) Stock Nitrate Solution : Dry pottasium nitrate in an oven 105 degree centigrade for 24 hours. Dissolve
0.7128g in water and dilute to 100ml. 2) Intermediate
Nitrate Solution : Dilute 100ml of stock nitrate solution to 1000ml with d.w. 3) From intermediate
nitrate solution prepare standards of 1,2,3,4,5,6 and 7 ppm. Treat them same as sample.
Reagents: 4) Hydrochloric Acid 1N : 83ml
diluted to 1000ml d.w

Absorbing Media: N/A

Interference : N/A

Standardization N/A
Sr. No. PROCEDURE RESPONSIBILITY
50ml of clear sample add 1ml of HCL solution and mix thoroughly and read the
1 absorbance at 220 nm using distilled water as ab blank. Give charcoal treatment JSO/JSA
for coloured samples.

Calculation :- N/A

Reference :- APHA 19th Edition

Date Of Change Description of Change Prepared By Reviewed By Approved By

 WI No.
NITRATE Rev. No. 0
Date 10.12.2013
Method : Argentometric Method

Principle : In a neutral or slightly alkaline solution, potassium chromate (K20r04) can indicate the end point of the
silver nitrate titration of chloride. Silver chloride is ppt quantitatively before red silver chromate is formed.

Apparatus : N/A

1) Potassium Chromate Indicator Solution : Dissolve 50 gms K2Cra4 in little d.w. A( AgNO3 solution
until a definite red ppt is form( Stand for 12 hr, fi1ter and dilute to I R. with d.w. 2) Std. Silver
Nitrate solution 0.0141N : Dissolve 2.395 gms AgNO3 in d\st water and dilute to 1000 mi. Store in th
bottle.
3) Std. Sodium Chloride solution 0.0141N : Dissolve 824.0mg NaCI (dried at 140°C) in distilled water
Reagents: and dilute to 1000m1. 4) Special
Reagent for removal of interference : Dissolve 125gms Aluminium pot. sulfate or aluminium ammonium
sulfate AIK (S04)2 .12 H20 or AINH4 (S04)2 .12 H20 in I lit. distilled water. Warm to 60°C and add 55ml
conc. M-140H slowly with stirring. Stand for 1 hr., transfer to a large bottle and wash ppt by successive
additions with thorough mixing and decanting with d.w. until free from chloride. When freshly prepared the
suspension occupies a volume of 1 Lit.

Absorbing Media: N/A

Interference : Sulfide, Thiosulfate, Sulfite ions interfere but can be removed by treatment with hydrogen peroxide.

Titration : Titrate sample in the pH range 7 - 10 . Adjust sample pH to 7-10 with H2SO4 or NaOH . If it is not in this
range. Add few drops K2Cr2O7 indicator soilution.

Sr. No. PROCEDURE RESPONSIBILITY

Take 100ml sample. If the sample is highly coloured, add 3 ml Al(OH)3 solution.
1 Mix and allow to settle 4 filter. If the sulfide, sulfite, Thiosulfate, present add 1ml JSO/JSA
H2O2 and stir for 1 min.

mg Cl/L = Burette reading x N x 35,450 ml

Calculation :- of sample

N = Normality of AgNO3 (0.0141N)

Reference :- APHA 19th Edition

Date Of Change Description of Change Prepared By Reviewed By Approved By

 WI No.
COLOUR Rev. No. 0
Date 10.12.2013
Method : N/A

For a given wavelength, the transmission of the liquid is propotional to its concentration and thickness
Principle : through which the observer looks. Hence the colour of the liquid is half when viewed in half the thickness or
i.e. monochromatic light.

Reagents: N/A

Apparatus : N/A

Absorbing Media: N/A

Maintenance : To maintain performance of the Tintometer model F. Spillages on the instrument or in the sample chamber
should be cleaned immediately. Coloured glass filter should be kept as clean as possible.

Sr. No. PROCEDURE RESPONSIBILITY

Place the sample in cuvette. Focuss the viewing tube until a sharp image of the
1 apparatus is obtained.

2 Slides the tabs comtrolling the coloured filters to the right. JSO/JSA

The proportion of Red, Yellow, Blue must be adjusted until a coloyr match is
3
obtained.

Calculation : N/A

Reference: Lovibond Tintometer manual of Colour

Date Of Change Description of Change Prepared By Reviewed By Approved By

 WI No.
CONDUCTIVITY Rev. No. 0
Date 10.12.2013
Conductivit is the measurement of materials ability to conduct electric current. The basic unit of
Theory : conductance G is 'Siemens' . Conductance is reciprocal of resistance.

Measurement of conductivity of sampple solution gives the degree of electrical conductance in a defined
volume of solution. Electrical conductivity of the solution is propotional to the no. of ions available in it.
Principle : Hence conductivity measurement will give direct reading of the solution concentration. The principle by
which the instrument measures conductivity is two plates are placed in the sample and a potential applied
across them and then the current get measured.

Standard potassium chloride solution - KCL(0.0100 M) . Dissolve 745.6mg anhydrous KCL in conductivity
Requirements : water and dilute to 1000 ml by D/W in volumetric flask at 25 ̊C.

Standardization: Calibration of conductivity meter by 0.0100 M solution.

Standardization instrument daily 0.0100 M KCL. Wash electrode properly by D/W and clean by tissue
Quality Control : paper.

Unit: μS/cm i.e., microsiemens per centimeter.

Sr. No. PROCEDURE RESPONSIBILITY

Take 100 ml sample. Measure the conductivity of sample by conductivity

1 electrode.
JSO/JSA

2 Wash the electrode with distilled water properly during each sample.

Calculation : N/A

Reference: LabIndia manual of conductivity

Date Of Change Description of Change Prepared By Reviewed By Approved By

 WI No.
CYANIDE Rev. No. 0
Date 10.12.2013
Method : Colorimetric - Bispyrazolone Method

Cyanide in the alkaline distillate is converted to CNCl by reaction with Chloramines – T at pH < 8 without
hydrolyzing to CNO. After the reaction is complete CNCl forms a red blue colour on addition of a per
Principle : iodine – barbituric acid regent. Maximum colour absorption is between 575 and 582 nm .

1) Chlorammine T : Dissolve 01 gms in 100 ml D.W. 2)

Bispyrazolone reagent : Dissolve 0.2 g bispyrazolone in 20ml pyridine. Prepare fresh daily.
3) 3 Methyl 1 Phenyl 5 Pyrazolone reagent : Dissolve 0.02 g of 3 methyl 1 phenyl 5 pyrazolone in 250 ml
Reagents: mixture of 125 ml absolute ethyl alcohol add 125ml distilled water. 4) Mix
reagent : Mix 20 ml of bispyrazolone reagent with 80ml 3 methyl 1 phenyl 5 pyrazolone reagent. 5) a)
Stock cyanide solution : Dissolve 2.51g potassium cyanide in 1000ml d.w. This gives 1000 ppm solution.
b) Std. Cyanide
solution : Dilute 10ml of stock cyanide solution to 1000ml with distilled water.This gives 10 ppm solution.
c) Dilute std. Cyanide solution
further to get cyanide solution of 0.2,0.4,0.6,0.8 and 1.0 ppm concentrations.

Standardisation : N/A

Apparatus : N/A

Absorbing Media: N/A

Interferance : N/A

Sr. No. PROCEDURE RESPONSIBILITY

Take 100 ml sample. Add 0.2 g of sulphuric acid. Then add 4 ml MgCl 2. Then add
1 10 ml 1:1 H2SO4. Collect the distillate in 5 ml 1N NaOH. Collect 50ml distillate.
Take 25 ml of distillate for analysis.

2 JSO/JSA
Adjust pH between 6 and 7 with glacial acetic acid. Add 1ml of
chloramines T reagent and mix.

3 Add 5ml of mix reagent and mix. Wait for 40 min. and take absorbance at
620nm.

Calculation : mg/Liter of Cyanide = mg/Liter of cyanide from graph x Dilution Factor

Reference: Water APHA 19th Edition

Date Of Change Description of Change Prepared By Reviewed By Approved By

 WI No.
SALINITY Rev. No. 0
Date 10.12.2013
Method : 210 C. Argentometric Method

Principle : N/A
1) Silver nitrate solution (0 28 N) - Dissolve 48.5 gm AgNO3 in 500ml distilled water and dilute to 1000
ml. Store in glass stoppered brown glass bottle at room temperature. 2)
Reagents: Potassium chromate indicator solution: - Dissolve 63 gm K2Cr2O7 in 100ml distilled water. Add a few
drops of 0.28N AgNO3, until a definite red precipitate persists . Let stand to settle, filter and store in glass
dropping bottle. 3) Std Sodium chloride:
- Dry about 35 gm NaCI to constant weight. Cool and weigh out 29.674 gm. Dissolve in distilled water and
dilute to 1000 ml

Place 25 ml standard NaCl solution in a 150 ml Erlenmeyer flask. Add 6 drops of chromate indicator and
titrate with AgNO3 solution in yellow light until a red precipitate just forms. Stopper flask and shake
vigorously to break curds of AgCl. Continue titration to brown end point.
Standardisation : Normality = 12.69
ml AgNO3

Apparatus : N/A

Absorbing Media: N/A

Interferance : N/A

Sr. No. PROCEDURE RESPONSIBILITY

1 Use 25 ml sample and titrate as directed above JSO/JSA

1) Calculate the chlorosity equivalent of 1ml AgNO3 solution : Cleq = N x 0.0355

Cleq = Chlorosity equivalent N = Normality of AgNO3.
2) Calculate the chlorosity(Clo) = d x Cleq x 40

d = ml titrant used Cleq = chlorosity equivalent of AgNO3

Calculation : 3) Convert chlorosity to salinity by using Table 210 : IV.
OR
Calculate using : Sg/Kg = 0.03 + [ 1.805 x Chlorosity x 1/P20 ]
P20 is density of sea water

Reference: Water APHA 19th Edition

Date Of Change Description of Change Prepared By Reviewed By Approved By

 WI No.
NITRITE NITROGEN Rev. No. 0
Date 10.12.2013
Method : Colorimetric method using NEDA reagent
Principle : N/A

1. NEDA Reagent : Add 100ml phosphoric acid and 10g sulphanilamide. Dissolve it completely, add 1g of
Reagents: (1-napthyl)
2. Ethylenediamine dihrdrochloride : Mix and dissolve and then dilute to 1 lit with d.w. This solution is
stable for one month when stored in dark bottle in refridgerator.

a) Stock Nitrite Solution : Dissolve 1.232g NaNO2 in nitrite free water and dilute to 1000ml
b) Intermediate Nitrite Solution : 50ml of stock Nitrite solution and dilute to 250ml
c) Standard Nitrite Solution : Dilute 100ml of intermediate solution to 1000ml.
Standardisation : Prepare standards in volumetric flask by adding the followingg of Sodium Nitrite solution and dilute to 50ml.
Prepare standards of 0.05,0.1,0.2,0.3,0.4 and 0.6 ppm.

Apparatus : N/A
Absorbing Media: N/A
Interferance : N/A
Sr. No. PROCEDURE RESPONSIBILITY

50 ml clear sample, adjust the pH of the extract with 1M NaOH or 1M HCL in the
1 range of 5 to 9, add 2 ml od NEDA reagent.
JSO/JSA
Wait for minutes and read the absorbance at 543 nm. Give charcoal treatment
2 for coloured samples.

1) Calculate the chlorosity equivalent of 1ml AgNO3 solution : Cleq = N x 0.0355

Cleq = Chlorosity equivalent N = Normality of AgNO3.
2) Calculate the chlorosity(Clo) = d x Cleq x 40

d = ml titrant used Cleq = chlorosity equivalent of AgNO3

Calculation :
3) Convert chlorosity to salinity by using Table 210 : IV.
OR
Calculate using : Sg/Kg = 0.03 + [ 1.805 x Chlorosity x 1/P20 ]
P20 is density of sea water

Reference: NEERI

Date Of Change Description of Change Prepared By Reviewed By Approved By

 WI No.
PHOSPHATE Rev. No. 0
Date 10.12.2013
Method : ANSA method

Phosphate ion combines with ammonium molybdate under acidic conditions to form a complex ammonium
phosphomolybdate. Molybdenum in ammonium phosphomolybdate is readily reduced by amino-Naphthol
Principle : Sulfonic acid.

a) Ammonium Molybdate solution : Dissolve 31.4g of ammonium molybdate in about 200ml distilled water.
Add carefully 252ml conc. Sulphuric Acid to 400ml d.w, cool and add 3.4ml conc. Nitric acid, to this add
Reagents: molybdate solution and dilute to 1000ml.
b) 1-amino 2-naphtha 4-
Sulfonic Acid : Weigh out seperately 0.78g ANSA, 42g sodium sulphite and 70g sodium metabisulfite in
mortar. Mix ANSA with small proportion of sodium metabisulfite in mortar to solid state. Dissolve remaining
salts in small quantity of water. Dissolve ANSA mix in salt solution mix in solution. Dilute to 1000ml with
distilled water.

Dissolve 219.5mg potassium dihydrate phosphate in 1 lit. d.w.

Standardisation :

Apparatus : N/A

Absorbing Media: N/A

Interferance : N/A

Sr. No. PROCEDURE RESPONSIBILITY

Take 50ml sample, add 2ml ammonium molybdate and add 2ml ANSA. Then
1 wait for 20min, blue colour appears. Then read on spectrophotometer at
wavelength 690nm.

Intermediate standard solution : Take 10ml stock. Dilute it to 100ml. This JSO/JSA
2
gives 5ppm solution.

3 From this intermediate solution, prepare standard solutions of

0.2,0.4,0.6,0.8,1.0,1.2.1.4 ppm.

Calculation : N/A

Reference: Water APHA 12th Edition

Date Of Change Description of Change Prepared By Reviewed By Approved By

 WI No.
FLUORIDE Rev. No. 0
Date 10.12.2013
Method : SPADNS ZIRCONIUM (colorimetric)

The SPADNS colorimetric method is based on reaction between Fluoride and Zirconium dye lake. Fluoride
Principle : reacts with the dye lake, dissociating a portion of it into colourless complex anion and the dye.

a) SPADNS reagent : Dissolve 958mg SPADNS(Sodium 2- para Sulfo phenylazo)-1,8 dihydroxy-3

naphthalene disulphonate) in distilled water and dilute to 500ml.
Reagents: b)Zirconyl Acid reagent: Dissolve 133mg Zirconyl chloride Octahy in about 25ml distilled water. Add
350ml of conc. HCL_add d.w. to make up to 500ml.
c) Acid Zirconyl-SPADNS : Mix equal volumes of SPADNS solution and Zirconyl Acid Reagent.
d) Reference Solution : Add 10ml SPADNS solution 10 100ml d.w. Dilute 7ml conc. HCL to 10ml d.w. and
add to the diluted SPADNS solution. This solution is usd for setting the instrument reference point(zero).

Preparation of Dissolve 221mg of Sodium Fluoride in distilled water and diluted to 1000ml.
Stock Solution :
Preparation of
Standard Dilute 100ml of stock solution to 1000ml with d.w,i.e. 1ml = 10μg F. From this prepare a range of standards
Solution : of 0.2,0.4,0.6,0.8,1.0 and 1.2 ppm.

Absorbing Media: N/A

Interferance : N/A

Sr. No. PROCEDURE RESPONSIBILITY

Take 50ml of sample or a proportion of sample diluted to 50ml with

1
distilled water.

Give charcoal treatment for coloured samples. To 50ml sample, add 10ml
2 JSO/JSA
Acid Zirconyl SPANDS reagent.

3 Mix well and read at 570nm

Calculation : N/A

Reference: Water APHA 19th Edition

Date Of Change Description of Change Prepared By Reviewed By Approved By

 WI No.
PHENOL Rev. No. 0
Date 10.12.2013
Method : Chloroform Extraction Method

Principle : N/A

1) Phosphoric Acid :10 ml of Ortho Phosphoric Acid is diluted to 100 ml with

distilled water.
Reagents: 2) Buffer Solution : Dissolve 16.9 of NH4Cl in 143ml concentrated NH4OH and dilute to 250ml with d.w.

3) Stock Phenol Solution : Dissolve 1mg of Phenol in 1000ml d.w.

4) 4- Amino Antipyrene Solution : 2gm of 4-Amino Antipyrene is dissolved in 100ml distilled water.
5) Potassium Ferricyanide : 8 gm of potassium ferricyanide dissolved in 100ml distilled water.

a) Preparastion of Stock Solution : 1gm of Phenol is dissolved in d.w. and diluted to 1000ml with d.w. It
Standardisation : contains 1000ppm of phenol. b)
Intermediate Phenol Solution : Dilute 10ml of stock slution in d.w. to 1000ml .

Standard
Solution : Conc.In ppm ml solution intermediate
0.2 ppm 2ml-------->100ml
0.4 ppm 4ml-------->100ml
0.6 ppm 6ml ------->100ml
0.8 ppm 8ml-------->100ml
1.0 ppm 10ml------>100ml

Extraction to be done as per given procedure and O.D is read at 460nm.

Absorbing Media: N/A

Interferance : N/A

Sr. No. PROCEDURE RESPONSIBILITY

Distillation Step : 100ml of sample + 2 ml of buffer solution to lower the

pH to 4.0 add 0.6ml of Potassium Ferricyanide + 0.6ml of Amino
1 JSO/JSA
Antipyrene. Shake well extract with 25ml of chloroform and let chloroform
settle down. Filter the chloroform and read the absorbance at 460nm.

Calculation : N/A

Reference: Water APHA 19th Edition

Date Of Change Description of Change Prepared By Reviewed By Approved By

 WI No.
DETERGENT IN WATER Rev. No. 0
Date 10.12.2013
Method : Anionic Surfactants as MBAS

Methylene blue active substances (MBAS) bring about the transfer of methylene blue, a cationic dye, from
an aqueous solution into an immiscible organic liquid upon equilibration. This occurs through ion pair
Principle : formation by the MBAS anion and the methylene blue cation. The intensity of the resulting blue colour in
the organic phase is a measure of MBAS.

The method is applicable at MBAS concentration down to about 0.025 ppm. Minimum detectable quantity
Detecion Limit is about 101.tg MBAS (calculated as LAS)

1.Stock LAS solution: 1.00 g LAS ( Linear alkylbenzene sulphonate) diluted to 1 litre gives 1000 ppm
solution.
2. Standard LAS solution: Dilute 10 ml of stock to llitre. This gives 10 ppm solution.
3. Phenolphthalein indicator
4. NaOH 1N
Reagents: 5. H2SO4 1N and 6N
6. CHC13
7. Methylene blue reagent: Dissolve 100mg methylene blue in 100m1 water. Take 30m1 to 1000m1 flask +
500m1 water + 41m1 6N H2SO4 + 50 gm NaH2PO4.H20 Shake till dissolves. Dilute to 1 litre.
8. Wash solution: Add 41m1 6N H2SO4 to 500 ml water in llitre flask. Add 50 g NaH2PO4. H20 and shake
until dissolved. Dilute to 1 litre. 9. H202

Glassware and 1. Seperatory funnels 2 nos. 500ml.

2. Test tube
Instrument :
3. Spectrophotometer foor use at 652nm

Standardisation : N/A

Absorbing Media: N/A

Interferance : N/A
Sr. No. PROCEDURE RESPONSIBILITY

Prepare stock LAS solution of 1000ppm. For this, prepare standards of

0.5, 1, 2,3, 4, and 5ppm. Follow the extraction procedure for each
1 JSO/JSA
standard. Read the standards at 652 nm. Plot a calibration curve of
absorbance vs ppm of standards.

Calculation : We get reading in ppm. Multiply by diluting factor in case any dilution of the sample is taken

Reference: Water APHA 19th Edition

Date Of Change Description of Change Prepared By Reviewed By Approved By

 HEXAVALENT
CHROMIUM
Method : DIPHENYLCARBAZIDE METHOD
Cr+6 reacts with diphenylcarbazide to produce a redish
Principle : purple color In slightly acid solution.MDC (minimum detection concentration) :-3µg/liter.
1) 1-5 diphenylcarbazide reagent - Dissolve 200 mg 1-5 diphenylcarbazide in 100ml 95% ethyl alcoh
or lsopropyl alcohol, add with mixing, an acid solution prepared from 40ml conc. H2SO4 and 360 ml
distilled water. Refrigerate to maintain stability for about a month. A color change from colorless to tan
does not affect the reagent’s usefulness
2) Stock
Reagents: Chromium Solution – Dissolve 141.4 mg anhydrous Potassium dichromate, K2Cr2O7, in distilled wate
and dilute to 1000ml : 1ml = 50 µg Cr.
3) Standard Chromium Solution
Dilute 10ml of stock chromium solution to 100ml. 1ml = 5μg of Cr. Prepare standards as :-
0.2,0.4,0.6,0.8,1.0 ppm from standard chromium solution.

Standardisation : N/A

Absorbing Media: N/A

Interferance : N/A
Sr. No. PROCEDURE

1 Use a 50 ml sample, if necessary, clarify by centrifuging.

Add 2.5 ml diphenylcarbazide reagent and mix well, take photometric

2 measurement at 540 µm with a 5 cm light path ; at least 5 min, but not
later than 15 min, after the reagent is added.

Calculation : N/A

Reference: Water APHA 13th Edition

Date Of Change Description of Change Prepared By Reviewed By

 WI No.
Rev. No. 0
Date 10.12.2013

ration) :-3µg/liter.
rbazide in 100ml 95% ethyl alcohol
0ml conc. H2SO4 and 360 ml
olor change from colorless to tan

2) Stock
omate, K2Cr2O7, in distilled water

3) Standard Chromium Solution :

pare standards as :-

RESPONSIBILITY

JSO/JSA

Approved By
 WI No.
SODIUM / POTASSIUM Rev. No. 0
Date 10.12.2013
Method : Flame emission photometric method

Trace amount of Na or K can be determined by flame emission at a wavelength of 589 nm & 766.5 nm.
The sample is sprayed into a gas flame & excitation is carried out under carefully controlled condition &
Principle : reproducibe condition. The intensity of light is measured by phototube potentiometer or other appropriate
circuit. The intensity of light at specific wavelenth is approximately propotional to concentration of the
element.

1) Sodium Chloride : For Na

Reagent : 2) Potassium Chloride :
For K
1) Stock Sodium Solution : Dissolve 2542g NaCl dried at 140 ̊C & dilute to 7 lit with d.w.
Intermediate Sodium Solution : Dilute 10ml stock sodium solution with d.q to 1000ml. Using
intermediate solution prepare standards as :- Lower range : 1,2,..... upto 10ppm. Higher Range :
10,20,....100ppm
Standardisation : 2) Stock Potassium Solution : Dissolve 1.907g KCl dried at 110 ̊C dilute to 1 lit with d.w. Intermediate
Potassium Solution : - Dilute 10ml stock potassium solution with distilled water to 1000 ml. Using
intermediate solution prepare standards as Lower range : 1,2.....upto 100ppm Higher Range :
10,20..100ppm

Absorbing Media: N/A

Interferance : N/A
Sr. No. PROCEDURE RESPONSIBILITY

Take samples and aspirate the sampe into the photometer using suction
1
tubes and compressor.
JSO/JSA
Measure the constant absorbance reading by using this reading find out
2
conc. Of Na/K

Calculation : N/A

Reference: Instrument mannual of Toshinwal

Date Of Change Description of Change Prepared By Reviewed By Approved By

 DETERMINATION OF WI No.
Rev. No. 0
MERCURY Date 10.12.2013
Method : Atomic Absorbtion Spectrometric Method usinf FIMS

The hydride technique involves the reaction pf acidified aquous samples with a reducing agent such as
sodium borohydride. The sodium borohydride reduction generates hydrides. The reaction generates a
Principle : volatile hydride which is transported to a quartz cell by means of an Argon carrier gas. In quartz cell the
hydrides are converted to gaseous metal atoms. These atoms are measured by the amount of light
absorbed.

a) 3% HCl : 30ml HCl diluted too 1000ml d.w.

b) 0.2% NaBH4 : 2g NaBH4 is dissolved in 0.05% NaOH solution ---> 1 lit.
c)K2S2O8 solution : Add 25g of potassium persulphate in 500ml d.w. heat at 40 ̊C untill dissolves.
d) HNO3 Solution : 500ml of conc. HNO3 to 400ml d.w. and then dilute to 1 lit.
Reagent :
e) 0.5N H2SO4 : 14ml of H2SO4 diluted to 1 lit of d.w.
f) KMnO4 : Dissolve 25g of KMnO4 to 500ml of d.w.
g) Sodium Chloride Hydroxylamine hydrochloride solution : Dissolve 60g of NaCl add 120g of
Hydroxylamine Hydrochloride in d.w. and dilute to 500mml.

1) Stock Solution : 1000ppm readymade stock solution is used for preparing working standards.
2) Intermediate Solution : Take 1 ml of stock solutiion dilute to 100ml, it gives 10ppm solution.
Standardisation : 3) Working standard solution : 0.1 ml of 10ppm --->100ml with 3% HCL----->10ppb
0.2 ml of 10ppm ----> 100ml with 3% HCL------>20ppb
0.3 ml of 10ppm -----> 100ml with 3% HCl -----> 30ppb

Absorbing Media: N/A

Interferance : N/A
Sr. No. PROCEDURE RESPONSIBILITY

1 The digested samples are run on FIMS toget reading in ppb for Hg. JSO/JSA

Calculation : N/A
1. EPA Method 245.1, In : Methods of determination of metals in environment samples supplement I, may
1994, EPA/600/R-94/111.
Reference:
2. Perking-Elmer FIMS setting up and performing analysis, Part number 0993-5203, The Perkin-Elmer
Cooperation, 76 main ave, Norwalk,(T06859(1994))
Date Of Change Description of Change Prepared By Reviewed By Approved By
 DETERMINATION OF WI No.
Rev. No. 0
METALS BY AAS Date 10.12.2013
Method : Atomic Absorption Spectroscopy

In this digested sample is aspirated into a flame and atimized. A light beam is diverted through the flame,
into a monochromator and onto a detector, that measures the amount of light absorbed by the atomized
Principle : element in the flame. The amount of energy at the characteristic wave length absorbed in the flame is
proportion to the conc. of th element in the sample.

1. Nitric Acid Conc HNO3 sample digetsion : Take 100ml sample in the beaker and add 5ml conc.
HNO3, and a few boiling chips. Boil it slowly and evaporate it to lowest volume possible before precipitation
occurs. Boil and add HNO3 if necessary to get clear soln. Wash down flask walls with water and then filter
Reagent : if necessary. Transfer the filterate to a 100ml volumetric flask with two 5ml portion of water. adding these to
volumetric flask. Take portion of this sooln required for metal determination.

1) Stock Solution : 1000ppm readymade stock solution for each metal can be prepared.
2) Standard Solution : They are prepared from stock soln in a given range.
Metals Range in which standards can be prepared
1) Copper 1-5 ppm AAS/ICP
2) Nickle 1-5 ppm AAS/ICP
3) Cadmium 1-20 ppm AAS/ICP
4) Zinc 1-20 ppm AAS/ICP
5) Iron 1-20 ppm AAS/ICP
Standardisation : 6) Cobalt 1-20 ppm AAS/ICP
7) Chromium 1-20 ppm AAS/ICP
8) Aluminium 1-20 ppm AAS/ICP
9) Manganese 1-20 ppm AAS/ICP
10) Boron 1-20 ppm AAS/ICP
11) Arsenic 1-20 ppm AAS/ICP

Absorbing Media: N/A

Interferance : N/A
Sr. No. PROCEDURE RESPONSIBILITY

The digested samples are run on AAS/ICP to get reading in ppb for for
1 JSO/JSA
required metals

Calculation : N/A

Reference: 1) Standard Method for the examination of water waste(APHA) 19th edition , 1995.

Date Of Change Description of Change Prepared By Reviewed By Approved By

 WI No.
BIOCHEMICAL OXYGEN Rev. No. 0
DEMAND ( B. O. D. ) Date 10.12.2013
Method : Azide Modification
A biochemical oxygen demand (B.O.D.) test based on mainly bio-assay procedure which measures the
Principle : dissolved oxygen consumed by bio organisms while assimilating and oxidizing the organic matter under
aerobic conditions.

1) Phosphate Buffer Solution :-

Dissolved 8.5g potassium dihydrogen phosphate (KH2PO4) 21.75g dipotassium hydrogen phosphatge
(KH2PO4), 33.4g disodium hydrogen phosphate (Na2HPO4) 1.78gm ammonium chloride in about 500ml
distilled water and dilute to 1 lit. pH of the solution should be around 7.2 without any further adjustment.

2) Magnesium Sulphate Solution :-

Dissolve 22.5g magnesium sulphate (MgSO4.7H2O) in distilled water and dilute to 1 Lit.

3) Calcium Chloride Solution :-

Reagents :
Dissolve 27.5g calcium chloride in distilled water and dilute to 1 Lit.

4) Ferric Chloride Solution :-

Dissolve 0.25g hydrated ferric chloride (FeCl3.6H2O) in distilled water and dilute to 1 Lit.

5) Acid & Alkali Solution :-

1N H2SO4 (28ml conc. 1Lit) with D W 1N NaOH (40gm 1Lit) with D W For neutralization of samples.

Reagents For 1) Maganous :-

Dissolved Oxygen :
(MnCl2.4H2O) Dissolve manganese chloride 1000gms in 2.5 Lit. Chloride distilled water.

2) Azide Solution :-

Dissolve 500gms of sodium hydroxide (NaOH) in 750ml of distilled water and add 150gms of potassium
iodide to it Cool the solution I Solution. Dissolve 10 gm of sodium azide in 50ml of distilled water II
Solution. Mix solution I and II and dilute to 1 Lit.

3) Concentrated Sulphuric Acid :- (H2SO4)

4) Starch Indicator :-

Dissolve 10gm of starch in 500ml of hot distilled water. Add 2gm of salicylic acid as preservative.

5) Stock Sodium Thio Sulphate Solution (N/8) :-

(Na2S2O3.5H2O) Dissolve 31.02gm sodium thiosulphate in boiled distilled water and make up to 1000ml.
Add 1gm of sodium hydroxide to preserve it (0.125N).

6) Working Sodium Thiosulphate Solution :-

Dissolve 100ml of stock solution to 1 Lit. of distilled water (0.0125).

Standardization of Potassium Dichromate (K2Cr2O7);(N/80) : Dissolve 0.613gm of K2Cr2O7 in 1Lit. Of Distilled

Sodium water(0.0125N). Take 10ml of N/80 K2Cr2O7 and add approx. 50ml d.w. + 1ml conc H2SO4 to it, then add
Thiosulphate pich of KI. Keep it in dark place for 5 mins. Titrate the brown solution till pale yellow colour appears using
Solution : N/80 sodium thiosulphate solution. Then titrate with working sodium thiosulphate solution N/80 usinng
starch indicator(0.5ml). Blue to colourless end point.
Determination of BOD
Standardization;Glucose : Dry grade glucose and glutamic acid at 103 ̊ C for 1 hr. Add 150mg of glucose
and 150 mg of glutamic acid to distilled water and dilute to 1 lit.Prepare fresh immediately before use. For
periodical checking use a mix of 150mg of glucose and 150mg of glutamic acid per lit. as a standard.
Determine the 3 days 27 ̊ C BOD of 2 % dilution of the glucose - glutamic acid. If the BOD value is more
than 200±377 mg/Lit. reject the BOD determinaton mode with the seed.

Preparation of dilution water : Aerate the required volume of distilled water in a

container by bubbling compressed air for 8 to 12 hrs. to attain dissolved oxygen saturation. Let it stabilize
for 4 hrs. at room temp. around 270C for 3 days. At the time of use add 1ml each of phosphate buffer,
magnesium sulphate, calcium chloride and ferric chloride for each one Lit. of dilution water.

Absorbing Media: N/A

Interface : N/A

Standardization N/A

Sr. No. PROCEDURE RESPONSIBILITY

Taking sample for BOD determination, take DO of the sample.
Dissolved Oxygen (DO) : Take sample by shaking it properly in 125ml bottle.
1 Add 1ml MnCl2 + 1ml azide solution; PPt(Pricipetate) Forms shake the bottle and
ppt settles. Stopper the bottle. Dissolve ppt by adding 1ml conc. H 2SO4.

Titrate 100ml of the solution using sodium thiosulphate (N/80) solution and starch
indicator End point. Blue to colourless. Then take requisite quantity of sample in
2 500ml volumetric flask and dilute to 500ml; by dilution water. (Neturalize the JSO/JSA
sample to pH around 7.0 by using alkali or acid).

Determine the pH by bromothymol blue indicator. Fill the solution in 300ml BOD
bottle and 125ml DO bottle. Determine the initial DO and keep 300ml BOD bottle
3 for incubation at 270C + 10C for 3 days. Initial DO : 1ml MnCl2 + 1ml azide, shake
well settle ppt. Add 1ml conc. H2SO4. Titrate against N/80 Na2S2O3.5H20.

Initial DO : 1ml MnCL2 + 1ml azide, shake well settle ppt.

Add 1ml conc. H2SO4. Titrate against N/80 Na2S2O3.5H2O.

Final DO (after 3 2ml MnCl2 + 2ml azide. Settle ppt. Add 2ml conc. H2SO4.
days) : Titrate against N/80 Na2S2O3.5H2O.

Calculation :- BOD = B.R. (Difference.Bet’n IDO-FDO) X 0.0125 X 8 X 1000

Vol. Of sample taken
= 1 X 5 (since sample is 100,l diluted to 500ml)
= mg/lt.
Date Of Change Description of Change Prepared By Reviewed By Approved By
 WI No.
TOTAL RESIDUAL Rev. No. 0
CHLORINE (T.R.C.) Date 10.12.2013
Method : Titrimetric Method
Chlorine will liberate free iodine from Kl solution at pH 8 or less. The liberated iodine is titrated with a
Principle : standard solution of Na2S2O3. With starch as the indicator. Titrate at pH 3 to 4 because the reaction is not
stoichiometric at neutral pH due to partial oxidation of thiosulphate to sulfate.

Apparatus : N/A
Reagents: Glacial acetic acid, potassium iodide, starch solution sodium thiosulphate (0.0125 N) Solution.

Absorbing Media: N/A

Interface : N/A
Standardization N/A
Sr. No. PROCEDURE RESPONSIBILITY

Take 100ml sample + 1 ml Glacial Acetic Acid. pH should be between 3-4.

1

2 If sample is highly alkaline add required amount of Glacial Acetic Acid.

JSO/JSA
Pinch of Kl, keep in dark for 5 min., add starch.
3

Titrate with N/80 Na2S2O3 end point blue to colourless.

4

Calculation :- TRC mg/l (mg Cl as Cl2/l) = B.R. x N x 35450

ml of sample
=4.43 x BR (Burette reading)
N = Normality of Na2S2O3

Date Of Change Description of Change Prepared By Reviewed By Approved By

 WI No.
SILICA Rev. No. 0
Date 10.12.2013

Method : Molybdosilicate method.

Ammonium molybdate at approximate pH 1.2 reacts with silica and also with phosphate present, to
produce heteropoly acids. Oxalic acid is added to destroy the molybdo-phosporic acid but not
Principle : molybdosilicic acid. The intensity of the yellow colour is proportional to the conc. of molybdate reactive
silica.

Apparatus : N/A
1) Hydrochloric Acid : 1 + 1
2) Ammonium Molybdate : Dissolve 10 gm ammonium molybdate in distilled water with stirring and gentle
warming, dilute to 100m1. Adjust pH 7-8 with ammonium or sodium hydroxide and store in polythene bottle.

3) Oxalic Acid : Dissolve 10 gm of oxalic acid in distilled water and dilute to 100 ml.
4) Stock Silica Silution : Dissolve 4.73 gm Sodium metasilicate in boiled and cooled distilled water dilute
to 1000 ml. ca ▪ Dilute 10 ml of stock silica solution to 1000 ml.
5) Working Silica Solution(SiO2) : Dilute 10ml of stock solution to 1000ml with distilled water.
6) Prepare stds from 5 ppm to 20 ppm.

Absorbing Media: N/A

Interface : N/A
Standardization N/A
Sr. No. PROCEDURE RESPONSIBILITY

Take 50ml sample, add 1ml 1+1 HCl and 2ml ammonium molybdate reagent mix
1
by inverting at least six times.

2 Wait for 5-10 minutes. Then add 1.5ml oxalic acid and mix. JSO/JSA

3 Read colour after 2 min but before 15min. At ʎ=410.

Calculation :- mg/lSiO2 = μgSiO2

ml sample
Reference :
APHA 19th Edition

Date Of Change Description of Change Prepared By Reviewed By Approved By

 WI No.
TOTAL KJELDAHL Rev. No. 0
NITROGEN (T.K.N) Date 10.12.2013

Method : Total Kjeldahl Method

In the presence of Sulphuric Acid, Potassium Sulfate and Cupric Sulfate catalyst, amino nitrogen of many
Principle : organic materilas is covered to ammonium. Free Ammonnia is also converted ammonium. After addition of
base, the ammonia is distilled from an alkaline medium and absorbed in boric acid.

Apparatus : N/A
1) Sodium Hydroxide 40% (NaOH) : 500 gm NaOH in 1250 ml distilled water.
2) Mixed Indicator :Dissolve 200 mg methyl red indicator in 100ml 95% ethyl
alcohol. Dissolve 100mg methylene blue in 50ml 95% ethyl alcohol combine the two solution.
3) Indicating Boric Acid (H3BO3) : Dissolved 20gm Boric Acid in distilled water and add 10ml mixed
indicator solution and dilute to 1 lit.
Reagents: 4) Sulfuric Acid 0.02 N (H2SO4) : 2.8ml of conc.H2SO4 dilute to 1 lit. This makes 0.1 N H2SO4, From
0.1 N H2SO4 take 200ml solution and dilute to 1 lit. This makes 0.02 N H2SO4.
5) 0.02 N Sodium Bicarbonate ((Na2CO3) : 1.060 gm Na2CO3 dried at 140 ̊C dilute to 1 lit. For
standardization of sulfuric acid.
6) Potassium Sulfate : 7 gm
7) Cupric Sulfate : 0.8 gm

Absorbing Media: N/A

Interface : N/A
Standardization N/A
Sr. No. PROCEDURE RESPONSIBILITY

1 Take 50ml sample, add 7gm Potassium Sulfate and 0.8gm Cupric Sulfate and add
10ml Conc. H2SO4, mix well.

2 Digestion will complete at 350 ̊C. After Digestion sample will remain upto 10ml,
cool it and add 75ml d.w.
JSO/JSA

3 Then add 50ml 40% NaOH, After distillation collect Ammonia in 50ml indicating
Boric Acid, Collect 150ml distillate.

4
Then titrate against 0.02 N H2SO4.

B.R. x 280
Mg/Lit Organic Nitrogren = ml of sample
Calculation :-
= B.R. x 5.6

Reference : APHA(Water) 19th Edition

Date Of Change Description of Change Prepared By Reviewed By Approved By

 WI No.
AMMONIA Rev. No. 0
Date 10.12.2013

Method : Total Kjeldahl Method

Free Ammonia nitrogen can be recovered when the distillation mixture is kept near pH 7.4. A phosphate
buffer is applied for the purpose of maintaining the required pH during the distillation process. When pH is
Principle : too high, certain organic compounds of nitrogen are converted to ammonia and when pH is too low, the
recovery of ammonia is in-complete.

Apparatus : N/A
1) Phosphate Buffer Solution : ( pH-7.4) Dissolve 14.3 gm Potassium dihydrogen phosphate (KH2PO4) .
Dilute to 1 Lit. with Ammonia free water.
Reagents: 2) Nessler Reagent : Dissolve 100g Mercuric Iodide and 70gm Potassium Iodide in small quantity of
distilled water and 160gm Sodium Hydroxide in 500m1. distilled water dilute to 1 lit.

Absorbing Media: N/A

Interface : N/A
Standardization
Stock Ammonia Solution : Dissolve 3.891g anhydrous NH4Cl, dried at 100 ̊C, dilute to 1 lit. Working
Amonia Solution : Dilute 10ml stock ammonia solution to 1 lit. with d.w.

Sr. No. PROCEDURE RESPONSIBILITY

1 Take 100ml sample. Add 10ml phosphate buffer. Collect 100ml distillation.

Add 1ml Nesslers reagent. Read at 425nm. This method used for colourless river
2 sample. JSO/JSA

Take 100ml sample, add 1ml Nesslers reagent and take reading on
3 spectrophotometer at wavelength 425nm.

% NH3 = free & saline ammonia x free NH3 factor

Calculation :- 100

Reference : APHA(Water) 12th Edition

Date Of Change Description of Change Prepared By Reviewed By Approved By

 WI No.
CHEMICAL OXYGEN Rev. No. 0
DEMAND ( C.O.D.) Date 10.12.2013
Method : Open Reflux-Titrimetric Method

Organic matter is oxidized by a boiling mix of chromic and sulphuric acids. A sample is refluxed in strong
acid solution with a known excess of Potassium Dichromate. After digestion, the remaining unreduced
Potassium Dichromate is titrated with Ferrous Ammonium Sulphate to determine the amount of Potassium
Principle : Dichromate consumed and the oxidizable organic matter is calculated in terms of oxygen equivalent.

1) Std. Pot. Dichromate Solution : (K2Cr2O7)

Dissolve 12.259gm K2Cr2O7 previously dried at 103oC
for 2 hrs. in distilled water and dilute to 1000 ml.

2) Sulphuric Acid Con.:

Add AgSO4 25 gm to 2.5 Lit Sulfuric Acid (H2SO4)

3) Ferroin Indicator :
Dissolve 1.485gm 1, 10- Phenanthroline Monohydrate and 695mg Ferrous Sulphate (FeSO4) H2O in
distilled water and dilute to 100 ml.

4) Std. Ferrous Ammonium Sulfate (FAS)-0.1N :

Reagents: Dissolve 39 gm fe(NH4)2 (SO4)2 , 6 H2O) in distilled water add 20 ml conc. H2SO4, cool and dilute to
1000ml.

5) Mercuric Sulfate : HgSO4, Crystal or Powder.

6) Sulphamic Acid : 1gm is to be added to 1 lit of

K2Cr2O7 Solution (0.25N)

7) Potassium Hydrogen Phthalate(KHP) : Lightly crush and then dry potassium hydrogen phthalate to
constant weight at 120 ̊C. Dissolve 425mg in distilled water and dilute to 1000ml. KHP has a theorotical
COD of 500μgO2/ml.

Principle : N/A

Absorbing Media: N/A

Interface : N/A

Standardization N/A

Sr. No. PROCEDURE RESPONSIBILITY

Add pinch of : HgSO4 to a reflux tube (to eliminate the halides interference), add
1 10ml of K2Cr2O7 and 20ml of sample (depending upon the nature of sample), add JSO/JSA
30ml conc, H2SO4, reflux for 2 hrs. at 150oC + 2oC.

Cool to room temperature and add 2 to 3 drops of ferroin indicator and titrate with
2 Ferrous Ammonium Sulphate (FAS). JSO/JSA

3 At End. Pt.- Blue green – Reddish Brown. JSO/JSA

COD mg/l = (A-B) X N X 8 X 1000

Calculation :- ml of Sample
A = ml FAS used for blank B = ml FAS used for sample N = Normality of FAS

Reference : APHA - 19th Edition

Date Of Change Description of Change Prepared By Reviewed By Approved By

 WI No.
BACTERIOLOGY Rev. No. 0
Date 10.12.2013
Method : Open Reflux-Titrimetric Method

The multiple tube fermentation method of coliform testing uses test tubes of broth media that limits the
growth of acid & gas forming coliform bacteria. In each tube is a special inverted Durham's tube that traps
gabs formed by the bacteria. In the first step
of presumptive test media is MacConckey broth & the second step or confirmed media brilliant green
Inoculating MPN lactose bile broth. Water sample has been
tubes : added to the test tubes and incubated for a minimum of 24°C hr. at 35°C. The coliform group comprises all
aerobic and facultative anaerobic negaive, non spore forming, rod shaped bacteria that ferment lactodr with
gas formation within 48hours at 35 ̊C.

Media to which special, selective components have been added. Such additives help identify the target
organism through color changes or other responses. The responses are brought about through metabolic
or enzymatic processes specific to the target organism. Results of the examination of replicate tubes and
Defined Substarte : distillation are reported in terms pf the most porbable number(MPN). This number based on certain
probability formulas is an estimate of the mean destiny of coliforms in the sample.

N/A

Absorbing Media: N/A

Interface : N/A

Standardization N/A

Sr. No. PROCEDURE RESPONSIBILITY

1 N/A JSO/JSA

MPN/100 ml = no of positive tubes x 100

Calculation :- sqrt(ml sample in negative tube) x ( ml samples in all tubes)

Reference : APHA - 16th Edition , STANDARD METHODS

Date Of Change Description of Change Prepared By Reviewed By Approved By

 WI No.
SULPHIDE Rev. No. 0
Date 10.12.2013
Method : N/A

1. Amine- Sulfuric acid stock soln : Weigh 13.5g of N-Ndimethyl-P-0Phenylene diamine oxalate. Take a mix
of 25ml conc. H2SO4 + 10ml d.w. in a beaker and add above rgt. Into beaker, By contionus stirring
dissolve all powder into the soln. For that cool the beaker under the tap water, when the powder form mix,
get totally dissolved, make it upto 50ml in volumetric flask by d.w. which then store in the dark glass bottle.

2. Amine sulphurinc acid rdt. : In a 2lit beaker, take 500ml d.w. in that add 500ml conc. H2SO4. By adding
H2SO4 put beaker it in cool water bath. From above amine sulphuric acid stock soln, take 12.5ml soln in
Reagent 500ml volumetric flask and add it upto 500ml by adding 1:1 H2SO4 store in dark glass bottle. This is our
Preparation : working reagent.

3. Ferric Chloride Solution : Weigh 100g of FeCl3.6H2O in 40ml d.w.

1. 0.1 N Sodium Thiosulphate : 12.5 gram of sodium thiosulphate and make it upto 500ml with d.w. 2.
0.1 N HCl : 0.83 ml of HCl make it 100ml by d.w. in 100ml volumetric flask. 3. 0.1
Reagents Required Standard Iodine Soln : 12.7g iodine + 40g of potssium iodide dissolve into d.w make it 1 lit. 4. Starch
for standardisation soln as an indicator : 10g of starch dissolves in 5000ml d.w. add 2g salicylic acid as preservative.
: 5. Stock sulfide soln : Weigh 4.10g
sodium sulfide trihydrate make it upto 1000ml by previouosly boiled, cooled d.w. in volumetric flask and
immediately stopped it.

Principle : N/A

Absorbing Media: N/A

Interface : N/A
Standardization N/A

Sr. No. PROCEDURE RESPONSIBILITY

Take 50ml sample + 1.5ml amine sulphuric acid rgt. + 0.5 ml Ferric chloride
1
stoppered the tube and take reading immediately at 600nm. On O.V
JSO/JSA
Read sample blank out adding reagents and subsequent from test reading which
2
will give you the final reading

STANDARDISATION PROCEDURE
Take 100ml d.w ion 250ml conical flask and add 0.1N 20ml standard iodine soln.
Add 25ml of 0.1N HCl , Dark orange red colour will appear titrate it with 0.1N
Na2S2O3.5H2O, untill the dark red orange color turns to yellow. After that add
1
starch indicator so it will appear as dark blue again titrate it with 0.1N
Na2S2O3.5H2O. The dark blue colour will appear as colourless. That reading is
marked as A. JSO/JSA
Take 100ml d.w in 250 ml conical flask. Addd 20ml 0.1N standard iodine solution
, add 15ml of 0.1N HCl and add 20ml of sodium sulfide soln. add starch indicator
2 and titrate it with sodium thiosulphide. End point will be dark blue to colourless.
That is marked as B.

Hydrogen Sulphide = [A -B ] x Normality of Thiosulphate x 17 x 1000

20
Calculation :-
A = Burette reading count of Hydrogen Sulphide B = Burette reading of hydrogen sulphide
17 = Sulphide molucular wt. 20 = ml of sample taken

Reference : APHA - 19th Edition

Date Of Change Description of Change Prepared By Reviewed By Approved By