PARAMETER – ACIDITY

METHOD – Titration Method

Principle - Acidity of water is its quantitative capacity to react with a strong base to a
designated pH. It may be defined as equivalent concentration of hydrogen ions in mg/l.
Sampling - Sample should be free from turbidity and if not filter through 0.45 / µm
membrane filter.
Apparatus – pH meter , Burette, Magnetic stirrer device.
Reagents -
Distilled Water - pH should not be less than 6·0. If the pH is less than 6.0, it shall be
freshly boiled for 15 minutes and cooled to room temperature. Deionized water may
be used provided that it has a conductance of less than 2 µs/cm and a pH more than
6·0.
Potassium acid phthalate solution, 0·02 N - Dissolve 4·0846 g of potassium acid
phthalate salt (dried at 120°C for 2 hours) in carbon dioxide free distilled water and
dilute to 1 L.
Sodium Hydroxide Solution - 15 N.
Sodium hydroxide solution -- 1 N Dilute 67 ml of 15 N sodium hydroxide solution to 1 L
With distilled water.
Sodium hydroxide solution – 0.02 N. Dilute 20 ml of 1 N sodium hydroxide solution to
1L and standardize using standard potassium acid phthalate.
Phenolphthalein Indicator - Dissolve 0.5 g of phenolphthalein in 100 ml, 1: 1 ( v/v) alcohol
water mixture and add 0.02 N sodium hydroxide solution drop by drop till very faint pink
colour is observed.
Methyl Orange Indicator - Dissolve 0·5 g of methyl orange in distilled water and make up to
100 ml in a volumetric flask.

Procedure –
Pipette 20 ml or a suitable aliquot of sample
into a 100-ml flask.

Determine the pH of water.

If pH is less than, 3.7, add two drops of methyl orange indicator into the first sample flask.

titrate with standard 0·02 N sodium hydroxide solution

until the colour changes to the faint orange.
 Record the volume of sodium hydroxide used

To the second sample beaker, add 2 to 3 drops

of phenolphthalein indicator .

titrate with 0·02 N sodium hydroxide solution to

the appearance of faint pink colour.

Record the volume used.

Calculation –
Acidity at pH 3.7, as mg/l CaC03 = A x N x 50 000
V
Acidity at pH 8.3, as mg/I CaC03 = B x N x 50000
V
where,
A= volume in ml of standard sodium hydroxide used to titrate to pH 3.7,
N =normality of standard sodium hydroxide,
V= volume in ml of sample taken for test, and
B= volume in ml of standard sodium hydroxide used to titrate to pH 8·3.

Reference - IS 3025 (Part 22).

PARAMETER - Sulfide
Method – Methylene Blue Method
Sampling - Collect water samples with minimum aeration. Either analyze samples
immediately after collection or preserve with zinc acetate solution for later analysis. To
preserve a sample for a total sulfide determination put zinc acetate and sodium
hydroxide solutions into sample bottle before filling it with sample.
Apparatus -
Matched test tubes, approximately 125 mm long and 15 mm OD.
Droppers, delivering 20 drops/mL methylene blue solution
Spectrophotometer, for use at a wavelength of 664 nm with cells providing light paths of
1 cm .
Reagents –
Amine-sulfuric acid stock solution: Dissolve 27 g N,N-dimethyl-p-phenylenediamine
oxalate in an iced mixture of 50 mL conc. H2SO4 and 20 mL distilled water. Cool and
dilute to 100 mL with distilled water.
Amine-sulfuric acid reagent: Dilute 25 mL amine-sulfuric acid stock solution with 975 mL
1 + 1 H2SO4. Store in a dark glass bottle.
Ferric chloride solution: Dissolve 100 g FeCl3⋅6H2O in 40 mL water.
Sulfuric acid solution, H2SO4, 1 : 1.
Diammonium hydrogen phosphate solution: Dissolve 400 g (NH4)2HPO4 in 800 mL
distilled water.
Methylene blue solution : Use USP grade dye or one certified by the Biological Stain.
Dissolve 1.0 g in distilled water and make up to 1 L.
Procedure –
Transfer 7.5 mL sample to each of two matched test tubes

If sample has been preserved with zinc acetate,

shake vigorously before taking subsample.

Add to Tube A 0.5 mL amine-sulfuric acid reagent

and 0.15 mL (3 drops) FeCl3 solution.
 Mix immediately by inverting slowly.

To Tube B add 0.5 mL 1 : 1 H2SO4 and 0.15 mL (3 drops) FeCl3 solution

And mix.

The presence of S2 will be indicated by the

appearance of blue color in Tube A.

Wait 3 to 5 min and add 1.6 mL (NH4)2HPO4 solution

to each tube.

Wait 3 to 15 min and make color comparisons.

 Zero instrument with a portion of treated
sample from tube B

And measure absorbance at 664 nm using 1 cm cell.

Calculation - Read sulfide concentration from calibration curve.

Limit – ETP – Inland surface water – max 2 mg/L, Marine – max 5 mg/L
Drinking water – 0.05 mg/L
Packaged drinking water – max 0.05 ppm
Natural packaged mineral water- max 0.05 ppm

Reference - APHA 4500

 PARAMETER - Bromide
Method – Phenol Red colorimetric method.
Principle - When a sample containing bromide ions (Br–) is treated with a dilute
solution of chloramine-T in the presence of phenol red, the oxidation of bromide and
subsequent bromination of the phenol red occur readily.
Apparatus –
Spectrophotometer, for use at 590 nm, providing 2cm cell
Nessler tubes, matched, 100-mL, tall form.
Acid-washed glassware: Wash all glassware with 1 + 6 HNO3 and rinse with distilled
water to remove all trace of adsorbed bromide.
Reagents -
Acetate buffer solution: Dissolve 90 g NaCl and 68 g sodium acetate trihydrate, in
distilled water. Add 30 mL conc. (glacial) acetic acid and make up to 1 L. The pH should
be 4.6 to 4.7.
Phenol red indicator solution: Dissolve 21 mg phenolsulfonephthalein sodium salt and
dilute to 100 mL with distilled water.
 Chloramine-T solution: Dissolve 500 mg chloramine-T, sodium p-
toluenesulfonchloramide, and dilute to 100 mL with distilled water. Store in a dark bottle
and refrigerate.
Sodium thiosulfate, 2M: Dissolve 49.6 g Na2S2O3⋅5H2O or 31.6 g Na2S2O3 and dilute to
1L.
Stock bromide solution: Dissolve 744.6 mg anhydrous KBr in distilled water and make up
to 1000 mL
Standard bromide solution: Dilute 10.00 mL stock bromide solution to 1000 mL with
water.
Procedure –
Add 2 mL buffer solution, 2 mL phenol red solution, and
0.5 mL chloramine-T solution to 50.0 mL sample.

Mix thoroughly immediately after each addition.

Exactly 20 min after adding chloramine-T, dechlorinate

by adding, with mixing, 0.5 mL Na2S2O3 solution.
 And read absorbance at 590nm.
Calculation –
mg Br/L = mg Br/L (from calibration curve) × dilution factor (if any)

Reference - APHA 4500 Bromide B

PARAMETER - ALKALINITY
Method – Titrimetric method
Principle - Alkalinity of water is the capacity of that water to accept protons. It may g be
defined as the quantitative capacity of an aqueous medium to react with hydrogen Ions
to pH 8.3 (phenolphthalein alkalinity) and then to pH 3.7 (total alkalinity or methyl
orange alkalinity).

Apparatus – 250ml conical flask, Burette

Reagents -
Distilled water
Sulfuric acid- 0.02 N
Mixed indicator- 0.02g methyl red and 0.01g bromocresol green in 95% ethanol and
make volume 100ml.
Procedure –
Take 20ml water sample in conical flask.

To this add 2 to 3 drops mixed indicator.

And titrate against 0.02N H2SO4.

End point green to pink.

Calculation -
Alkalinity as (as mg/l of CaCO3) = - A × N × 50000
V

Where,
A = volume of standard sulfuric acid required.
N = Normality of H2SO4 V = Vol. of sample taken
 PARAMETER - Chloride
Method – Titrimetric Method
Principle – In a neutral or slightly alkaline solution, potassium chromate can indicate the
End point of the silver nitrate titration of chloride. Silver chloride is precipitated before
Red silver chromate is formed.
Apparatus – Burette, Conical flask , Measuring cylinder.
Reagents -

Potassium chromate indicator- Dissolve 50 g potassium chromate in a little distilled

water. Add silver nitrate solution until a definite precipitate is formed. Let it stand for
12hr. Filter and dilute to 1L with water.

Silver nitrate (0.0141 N) - Dissolve 2.395 g silver nitrate in distilled water and dilute to1L

H2SO4 ( 1N ) , NaOH ( 1N ).
Procedure –
Take 50ml sample in conical flask.

Adjust sample of pH if it is in not range with

H2SO4 or NaOH.

Add 1.0ml potassium chromate indicator.

And titrate against standard silver nitrate to end point.

Standardize silver nitrate solution and conduct blank

using water as per same procedure.
Calculation –
Chloride mg/L = ( V1 – V2 ) × N × 35450
V3
Where,
V1 = Vol. of silver nitrate used for sample.
V2 = Vol. of silver nitrate used for blank.
V3 = Vol. of sample taken for titration.
N = Normality of silver nitrate solution.

Limit – Drinking water – max 250 mg/L and permissible 1000 mg/L
Packaged drinking water – 200ppm max
Natural packaged mineral water – 200 ppm max

Reference - IS 3025 (Part 32)

PARAMETER - Total Hardness
Method – Titrimetric method
Principle - This method depends on ability of ethylenediamine tetra acetic acid or its
disodium salt to form stable complexes with calcium and magnesium ions. When the
dye eriochrome black T (EBT) is added to a solution containing calcium and magnesium
ions at pH 10.0 a wine red complex is formed.
Apparatus – 250ml conical flask, Burette
Reagents -
Buffer solution – Dissolve 16.9g ammonium chloride in 143ml conc. Ammonium
hydroxide, add 1.25g magnesium salts of EDTA and dilute to 250ml with distilled water.
Dilute 10ml solution to 100ml.
Eriochrome black T indicator- Dissolve 0.4g EBT & 4.5 g hydroxylamine hydrochloride in
100ml 95% ethanol. This indicator is stable for 2months.
Standard EDTA- Dissolve 3.723g EDTA in demineralized water and make volume to
1000ml.
Std. Calcium solution - Weigh 1.000 g, suspend it in distilled water and add 1 : 1
hydrochloric acid AR quality, drop wise slowly to dissolve the solid. Use minimum
amount of acid. Boil for a few minutes, cool, add a few drops of methyl red indicator
and adjust to orange colour with 3 N ammonium hydroxide or 1 : 1 hydrochloric acid.
Dilute to 1 000 ml with distilled water.
Calculation –
Total Hardness as (as mg/l of CaCO3) = 1000 × (V1 – V2) × CF
V3
Where,
V1 = Vol. of EDTA used for sample
V2 = Vol. of EDTA used for blank
CF = Correction factor for standardization of EDTA
V3 = Vol. of sample
CF = X1/X2= X1 = volume in ml of standard calcium solution taken for standardization,
and X2 = volume of ml of EDTA solution used in the titration

Limit – Drinking water – max 200mg/L and permissible 600mg/L

Reference - IS 3025 (Part 21)

Procedure –
Take 50ml sample in conical flask.

To this add 2ml buffer solution.

Add 1 to 2 drops of EBT indicator.

And titrate against standard EDTA.

End point purple to blue.

Conduct blank same as above except sample use water.

PARAMETER – Fluoride
Method - Spectrophotometric Method
Principle - The SPANDS sodium 2-(parasulphophenylazo) 1,8-dihydroxy-3,6,
Naphthalene disulfonate, colorimetric method is based in the reaction between
fluoride and a zirconium dye lake. Fluoride reacts with dye lake, dissociating a portion
Of it into a colorless complex anion and dye. As the amount of fluoride increases, the
color become lighter.
Apparatus – Spectrophotometer, Glass apparatus
Reagents –
SPANDS – Dissolve 958 mg SPANDS sodium 2-(parasulphophenylazo) 1,8-dihydroxy-
3,6, Naphthalene disulfonate in distilled water and dilute to 500ml. Protect from direct
sunlight. This solution is stable for 1 year.
Zirconyl-acid reagent – Dissolve 133mg zirconyl chloride octahydrate in 25ml distilled
water. Add 350ml conc. HCL and dilute to 500ml with distilled water.
Acid-zirconyl SPANDS – Mix equal volumes of SPANDS solution and zirconyl acid
reagent. The combined reagent is stable for 2 years.
Sodium arsenite – 5g NaAsO2 and dilute to 1000ml with water.
Procedure –

Take 50ml sample in 100ml conical flask.

If the sample contains residual chlorine then

add 1 drop of sodium arsenite and mix.

To this add 5ml SPANDS reagent + 5ml acid- zirconyl reagent

or 10ml acid –zirconyl SPANDS reagent.

Mix well and read absorbance at 570 nm.

Calculation –
mg fluoride/ml = A × B
ml of sample C
A = mg of F- determined from plotted curve
B = final volume of diluted sample.
C = volume of diluted sample.

Limit – Drinking water - max 1 mg/L

ETP – Inland surface water- max 2 mg/L, Public sewers & marine –max 15mg/L
Packaged drinking water – 1.0 ppm max
Natural packaged mineral water - 1.0 ppm max

Reference - APHA 4500 F –

PARAMETER – Nitrite Nitrogen

METHOD – Spectrophotometric Method

Principle – Nitrite is determined by formation of a reddish purple azo dye produced at pH

2 to 2.5 by coupling diazotized sulfanilamide with N- (1-naphthyl)-ethylene- diamine
(NED) dihydrochloride. And absorbance is measured at 543 nm.

Reagents –
Nitrite free water – To 1L distilled water add small crystals of KMNo4 and calcium
hydroxide. Distill in an borosilicate glass apparatus and discard initial 50ml distillate. And
collect the distillate fraction.
Color Reagent – To 800ml water , add 100ml 85% phosphoric acid and 10g sulfanilamide.
After dissolving sulfanilamide completely, add 1g NED dihydrochloride, and dilute to 1L
with nitrite free water. Store it in dark bottle.
1N HCl, 1N NH4OH

Apparatus – Spectrophotometer for use at 543 nm, 1cm cell

Procedure –
Take 50ml solid free water sample.

Adjust pH of sample between 5 and 9.

And to this add sample 2ml color reagent and mix.

After 2hrs adding color reagent to samples

measure absorbance at 543nm.

Prepare standard curve by plotting of standard solutions.

Calculations – Compute sample concentration directly from curve (mg/L).
Limit – Packaged drinking water – 0.02 ppm max
Natural packaged mineral water – 0.02 ppm max

Reference – APHA 4500 23rd edition

PARAMETER – Sulfite
METHOD – Iodometric Method
Principle – In sulfuric acid solution by titration with potassium iodate solution against
starch indicator, iodide ions are oxidized to iodine, which in turn oxidizes sulfite ions to
sulfate ions. The sulfite concentration is determined from the consumption of titration
solution (iodometric determination).

Apparatus – Glass beaker, Conical flask, Burette

Reagents –
Sulfuric acid – 1:1
Standard potassium iodide- iodate titrant (0.002M) – Dissolve 0.4458 g anhydrous
KIO3+4.35g KI+310mg sodium bicarbonate in distilled water and dilute to 1000 ml
EDTA – Dissolve 2.5 g EDTA in 100ml distilled water
Starch indicator- To 5 g starch add 1L boiling distilled water, stir.
Procedure –
To a fresh sample add 1ml EDTA solution in 100ml sample

Cool/hot samples to 50°C. Do not filter.

Add 1ml H2SO4 and 0.1g sulfamic acid to suitable titration flask.

Add 1ml starch indicator solution to flask.

And immediately titrate with standard KI solution

Swirl flask, until faint blue color develops, analyze a reagent blank
Using distilled water instead of sample.
Calculations -
mg sulfite/L = (A - B) × M × 6 × 40000
ml of sample
Where,
A= mL titrant for sample,
B= mL titrant for blank,
M= Molarity of KI titrant.

Reference – APHA 4500 Sulfite 23rd edition

PARAMETER - NITRATE
Method - UV-VIS Spectrophotometer method
Principle – Sample is treated with 1M HCl and absorbance is measured at 220nm.
Because dissolved organic matter absorbs 220nm.

Apparatus – Spectrometer for use 220 nm and 275 nm, Quartz cuvette

Reagents –
Stock Nitrate solution – Dissolve 0.7218 g dried KNO3 in water and make volume
1000ml.
Intermediate stock solution – Dissolve 100ml stock to 1000ml water.
1M HCL – 83 ml Conc. HCL to 1000ml water
Procedure –
Take 50ml clear sample (filter if necessary)

Add 1ml 1M HCL and mix thoroughly

And read the absorbance at 220nm against reagent water.

And at 275nm to determine any interference

due to dissolved organic matter.
Calculation –
C = A- I
S
Where,
C = Concentration,
A = Absorbance,
I = Intercept
S = Slope
Limit – Drinking water- max 45mg/L
ETP – Inland surface water – 10mg/L, Marine water – 20mg/L
Packaged drinking water – 45 ppm max.
Natural packaged mineral water – 50 ppm max.

Reference – APHA 4500- NO3- 23rd edition

PARAMETER - SULPHATE
Method – Spectrophotometric method
Principle - Sulphate ion is precipitated in hydrochloric acid medium with barium chloride
in such a manner as to form barium sulphate crystals of uniform size. The absorbance of
barium sulphate suspension is measured by a spectrophotometer and the sulphate ion
concentration is determined by comparison of the reading with a standard curve.

Apparatus – Spectrophotometer, Glass cuvette, Glass apparatus

Reagents –
Conditioning reagent – Add 0.3 g gelatin powder in 100ml distilled water and warm it on
hot plate till it is dissolved. Gelatin solution is keep for overnight at 40C. After bringing
the solution to room temperature add 3g barium chloride is added to gelatin solution
and dissolve by mixing. Turbid solution is kept standing for 2hrs and mixed before use.
HCL (1+9).- Dissolve one volume of concentrated hydrochloric acid with 9Volumes of
distilled water.
Procedure-
Take 20ml clear aliquot of sample.

Add 1ml (1+9) HCL + 1ml conditioning reagent.

And mix well for 30 seconds.

And read the absorbance at 420nm.

Calculation –
Read the sulfate concentration directly from calibration curve. (mg/L)
Limit- Drinking water – max 200mg/L and permissible – 400 mg/L,
Packaged drinking water & Natural packaged mineral water – 200 ppm max.
Reference- IS 3025 ( Part 24 )
PARAMETER - AMMONIACAL NITROGEN
Method – By kjeldahl method
Principle- In the presence of H2SO4,potassium sulfate and cupric sulfate catalyst,
amino nitrogen of many organic materials converted to ammonium. Free ammonia is
also converted to ammonium. After addition of base, the ammonia Is distilled from an
alkaline medium and absorbed boric acid. And ammonia is determined by titration
method.
Apparatus- Kjeldahl flask capacity of 800ml, Heating mantel, Measuring cylinder,
distillation apparatus.
Reagent – Potassium sulfate, Copper sulfate, Conc. Sulfuric acid, Boric acid, Mixed
indicator, Sodium hydroxide, Sodium thiosulfate, Sodium tetraborate.
Borate buffer solution – Add 88ml 0.1N NaOH solution to 500ml approx.. 0.025M
sodium tetra borate and dilute to 1lit.
Dechlorinating reagent – Dissolve 3.5g sodium thiosulfate in water and dilute 1L.
Boric acid – Dissolve 20g boric acid in water add 10ml mixed indicator and dilute to
1000ml.
Mixed indicator- Dissolve 200mg methyl red in 100ml 95% ethanol + 100mg
methylene blue in 50ml 95% ethanol. Combine solutions.
Procedure –
Take 500ml dechlorinated water or portion or
portion of sample diluted to 500ml.

Remove residual chlorine by adding dechlorinating reagent.

Add 25ml borate buffer solution and adjust pH to 9.5

by adding 6N NaOH.

Connect distillation assembly.

Collect distillate into 500ml beaker containing 50ml boric acid solution.
 Collect at least 200ml distillate.

And titrate, distillate ammonia against standard sulfuric acid solution

And determine ammonia nitrogen.

Use residue in distilling flask for organic nitrogen.

For organic nitrogen determination, digest above sample.

Add 6.7ml conc. H2SO4 + 6.7g potassium sulfate + 0.365 g copper sulfate to distillation
flask
 Add few glass beads, and digest under a fuming hood.

Digest sample until volume is greatly reduced.

Continue digestion till solution becomes transparent and pale green.

After digestion, let cool, dilute to 300ml with water, and mix.

Connect flask to distillation apparatus.

Distill and collect 200ml distillate in the receiver containing 50ml boric acid.
 And titrate against standard acid solution.

Calculation -
Ammonical nitrogen mg/I= (A - B) × 280
V
Where
A= Volume in ml of sulfuric acid used for sample,
B= Volume in ml of sulfuric acid used for blank, and
V= Volume in ml of sample taken for test.
 PARAMETER - Oil & Grease
Method - Partition Gravimetric method.
Principle – Dissolved or emulsified oil & grease is extracted from water by
trichlorofluoroethane and estimation is gravimetrically.

Apparatus - Separating funnel 500ml capacity, Distillation flask,

Whatman filter paper No. 40 or equivalent, water bath, Glass funnel.
Reagent – Trichlorofluoroethane, Anhydrous sodium sulfate

Procedure –
Transfer 900ml sample to separating funnel.

Give washings to sample container with solvent

and transfer to separating funnel.

Shake vigorously for 2mins for 3 to 4 times

 Let it stand for separate layer.

Drain solvent layer through funnel containing filter paper with sodium sulfate in
weighed distillation flask.

Extract two more times with solvent and collect into distillation flask.

Give washings to filter paper with 10 to 20ml solvent.

Distill solvent from distillation flask.

And evaporate solvent on water bath at 700C.

 Keep flask in oven for removal of all solvent residue.

Cool in desiccator and weigh.

Calculation =
Oil & grease, mg/l = M × 1000
V
Where,
M = mass in mg of residue
V = volume in ml of sample taken.
Limits - ETP – Inland surface water (max)- 1o mg/L, Public sewers- 20 mg/L,
Marine – 20 mg/L

Reference - IS 3025 ( Part 39 )

PARAMETER - PHENOLIC COMPOUND

Method - By Spectrophotometric Method

Principle – Most phenols react with 4 –aminoantipyrine at pH 7.9 in the presence of
Potassium ferricyanide to form a coloured antipyrine dye. This dye is kept in aqueous
Solution and the absorbance is measured ta 460nm.
Apparatus – Spectrophotometer, pH meter
Reagent-
Ammonium Hydroxide - 0·5 N -Dilute 35 ml of fresh concentrated ammonium
hydroxide to 1 L with distilled water.
Phosphate Buffer Solution - Dissolve 104·5 g of potassium hydrogen phosphate and
72·3 g or potassium dihydrogen phosphate in distilled water and dilute to 1 L. The pH
of the solution should be 6·8.
4-Aminoantipyrine solution - Dissolve 2 g of 4-aminoantipyrine in distilled water and
dilute to 100 ml, Prepare daily.
Potassium Ferricyanide solution – Dissolve 8 g Potassium ferricyanide in distilled water
And dilute to 100ml. Stored in brown glass bottle.
Sodium sulfate anhydrous.
Procedure –
Place 100ml distillate or portion of sample diluted to 100ml

Add to this 12 ml of 0.5N ammonium hydroxide solution.

And adjust pH 7.9 with phosphate buffer.

Add 1ml 4-aminoantipyrine solution, mix well.

Add 1ml potassium ferricyanide solution, mix well.

Let it stand for 15mins and read absorbance of sample at 460nm.

Calculation – After obtaining absorbance values, depending upon the volume of
sample chosen for test, calculate the amount of phenol present in 1000ml as given
below,
Using calibration curve :
Phenol, mg/l = C × 1000
L
Where,
C= Concentration of phenol in mg in sample from the calibration curve
L = volume in ml of original sample.

Limits - Drinking water (max) – 0.001 mg/L and permissible 0.002 mg/L
ETP – Inland surface water – max 1 mg/L, Public sewers – 5 mg/L,
Marine water – 5 mg/L
Packaged drinking water & Natural packaged mineral water - Absent

Reference - IS: 3025 (Part 43)

PARAMETER – Phosphorous
Method – Vanadomolybdophosphoric acid colorimetric method
Principle - : In a dilute orthophosphate solution, ammonium molybdate reacts under
acid conditions to form a heteropoly acid, molybdophosphoric acid. In the presence of
vanadium, yellow vanadomolybdophosphoric acid is formed. The intensity of the
yellow color is proportional to phosphate concentration.
Apparatus – Spectrophotometer, Acid washed glassware, Filtration apparatus and
filter paper.
Reagents –
Phenolphthalein indicator aqueous solution.
Hydrochloric acid, HCl, 1 : 1.
Activated carbon
Vanadate- molybdate reagent:
1) Solution A: Dissolve 25 g ammonium molybdate, in 300 mL distilled water.
2) Solution B: Dissolve 1.25 g ammonium metavanadate, by heating to boiling in 300
mL distilled water. Cool and add 330 mL conc. HCl. Cool Solution B to room
temperature, pour Solution A into Solution B, mix, and dilute to 1 L.
Standard phosphate solution: Dissolve in distilled water 219.5 mg anhydrous KH2PO4 and
dilute to 1000 ml.
Procedure –
If sample pH is greater than 10, add 0.05 mL
(1 drop) phenolphthalein indicator to 50.0 mL sample.

discharge the red color with 1 + 1 HCl before diluting to 100 ml.

Remove excessive color in sample by shaking about 50 mL

with 200 mg activated carbon .

Place 35 mL or less of sample, containing 0.05 to 1.0 mg P,

in a 50-mL volumetric flask.
 Add 10 mL vanadate- molybdate reagent and
dilute to the mark with distilled water.

Prepare a blank in which 35 mL distilled water

is substituted for the sample.

After 10 min or more, measure absorbance of sample versus a blank

at a wavelength of 400 to 490 nm, depending on sensitivity desired.

Calculation –
mg P/L = mg P(in 50 mL final volume) × 1000
mL of Sample
 HEAVY METALS - Sample digestion
1. Cadmium - To 100ml acidified sample add 5ml conc. HCL and evaporate to 20 ml. Cool
and filter sample and make upto volume 100ml. Aspirate sample solution and measure
absorbance at 228.8 nm.
2. Zinc - Add 0.5ml nitric acid to 100ml sample. Add 5ml conc. HCl and filter sample
through acid washed filter paper. Make up volume 100ml. Aspirate solution and
measure absorbance at 213.8nm.
3. Nickel - Add 0.5ml nitric acid to suitable volume of sample in 100ml volumetric flask.
Make up to mark. Rinse nebulizer by aspirating water containing 1.5ml of conc. HNO3.
Aspirate sample solution and measure absorbance at 232nm.
4. Lead - To 100 ml acidified sample add 0.5ml nitric acid, 5ml conc. HCL and heat not to
boil but reduce volume 20ml. Cool and filter sample and make volume 100ml. Aspirate
sample solution and measure absorbance at 283.3 nm.
5. Copper - Add 5ml conc HCL to 100ml sample and evaporate solution to 15 to 20ml
Cool and filter sample through acid washed filter paper. Make up the volume 100 ml in
volumetric flask. Aspirate the solution and measure absorbance at 324.7 nm using
copper hollow-cathode lamp.
6. Aluminium - Add 0.5ml conc. nitric acid to 100ml sample in 250ml beaker. Add 5ml
conc. HCL. Heat on hot plate to reduce volume 50ml. And filter sample through 0.45
µm membrane filter. Transfer quantitatively to 100mlvolumetric flask and make
volume upto the mark. Aspirate sample solution and measure absorbance at 309.3nm.
7. Iron - Add 0.5ml conc. nitric acid to suitable volume of sample in 100ml volumetric flask.
Make upto the mark. Transfer contents to 150ml beaker. Add 25ml CaCl2 solution. Aspirate
sample solution and measure absorbance at 248.3 nm.
8. Chromium - Take 100ml sample, add 5ml conc. nitric acid, add few glass beads and heat
to boil solution and concentrate to lowest possible volume. Cool and transfer to beaker.
Add 5ml conc. nitric acid and 10ml conc. sulfuric acid. Heat until clear solution. Cool and
transfer to 100ml volumetric flask and make up volume 100ml. Add 1ml 30% hydrogen
peroxide. Aspirate the sample solution and measure absorbance at 357.9 nm.
9. Mercury - By cold vapor AAS - Transfer 100 ml sample to 300ml BOD sample. Add 5ml
conc. Sulfuric acid and 2.5ml conc. nitric acid. Add 15ml potassium permanganate. Let
stand for 15mins. Add 8ml potassium persulphate and 2hrs in water bath at 95℃. Cool and
add 6ml sodium chloride hydroxylamine solution to reduce excess permanganate. After
de-colourization add 5ml stannous chloride solution. And attach bottle to aeration
apparatus.
10. Arsenic- Take 50 ml sample. Add 1ml 2.5N sulfuric acid and 5ml 5% potassium
persulphate solution. Boil gently to reduce volume upto 10ml. Do not let go to sample
dryness. After manual digestion dilute 50ml. To 50ml digested sample add 5ml conc.HCL
and mix. Add 5ml sodium iodide pre-reductant sol.mix and wait at least for 30min. And
attach to reaction cell for producing arsenic hydride. And read concentration.
11. Selenium - To the sample add 5ml conc. Sulfuric acid + add 5ml hydrogen peroxide to
the RBF and add boiling beads. Connect flask to decomposition apparatus. And collect
condensate in condensate reservoir. Continue heating till fumes of sulfuric acid appear. If it
is turbid add another 5ml hydrogen peroxide and continue as per above. After cooling add
20ml HCL. Gently boil mixture under reflux for 15min. Cool the sample solution and
transfer to 100ml vol. flask. And make volume upto mark.