Total Alkalinity

Principles
The amount of acid required to titrate the base in the water is a measure
of alkalinity in the water. A number of bases including carbonates,
bicarbonates, hydroxides, silicates, phosphates, ammonia and various organic
compounds occurs in water. However carbonates and bicarbonates are
considered the predominant bases in natural waters. Since water above pH 4.5
may contains bicarbonates, water samples that turn yellow upon the addition of
methyl orange indicator contain alkalinity.
Apparatus
Pipettes, 10, 20 or 50 ml as required for aliquots
Burette, 10 or 25 ml, with 0.05 ml divisions
Beakers, 50 or 100ml
Erlenmeyer flasks, 100 or 200 ml
Graduated pipette, 1 ml, with 0.01 ml divisions
Reagents
Methyl orange indicator (in 1.0g of methyl orange add 100ml distilled water )
Sulphuric acid (0.10 N) 2.80ml H2SO4+ distilled water make volume upto
1000ml
Procedure
Total alkalinity was measured by methyl orange indicator method
(A.P.H.A., 1971). The samples for measurement of alkalinity were collected in
polyethylene bottles and were analyzed as soon as possible to avoid
denaturation. To a sub-sample of 50 ml in an Erlenmeyer’s flask, 0.1 ml
methyl orange indicator was added and titrated against 0.10 N standard
sulphuric acid to the end point (slight orange).
Calculation for estimation of total alkalinity was made by using the
following formula:

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 Volume of acid used x Normality of the acid x
50000
Total alkalinity (mg/1) = ____________________________________
Volume of the sample (ml)

Carbonates
Principles
When the pH value of a sample of natural water or water extract of a
soil is above 8.4, carbonate ion is present, normally as sodium carbonate. If the
sample is titrated with a standard mineral acid to a pH of 8.4, the carbonate ion
is converted to bicarbonate:
CO32- + H+ = HCO3–
and the amount of acid used is a measure of the carbonate present. The
titration is best carried out potantimetrically; but the end-point may also be
assessed by use of the mixed indicator cresol red-methyl blue, which changes
from violet at pH 8.8 to yellow at pH 8.0., with a distinctive rose-red hue at pH
8.4. The end-point is judged in the titration with indicator by matching the
colour to that of the same volume of buffer solution at pH 8.4 containing the
same amount of indicator.
Apparatus
Pipettes, 10, 20 or 50 ml as required for aliquots
Burette, 10 or 25 ml, with 0.05 ml divisions
Beakers, 50 or 100ml
Erlenmeyer flasks, 100 or 200 ml
Graduated pipette, 1 ml, with 0.01 ml divisions
Reagents
Phenolphthalein indicator (1.0g Phenolphthalein and add absolute
methanol upto 100ml)
Sulphuric acid (0.10 N)

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Procedure
To another sub-sample of 50 ml in an Erlenmeyer’s flask,
phenolphthalein (0.1 ml) was added as an indicator, then titrated against
sulphuric acid (0.10 N) until pink colour disappeared. The carbonates were
estimated by using the following formula:

Volume of the acid used x Normality of the acid x

50,000
Carbonates (mg/L) = _________________________________________
Volume of the sample (ml)

Bicarbonates
Principle
Bicarbonate ion reacts with acid and releases carbon dioxide into the solution:
HCO3– + H+ = H2O + CO2

The pH value at complete neutralization being about 3.8. Thus, bicarbonate

may be measured by titration with acid to a pH of 3.8, either potentometrically
or using an indicator unaffected by carbon dioxide. Methyl orange is suitable
and, if screened with a blue dye such as indigo carmine or xylene cyanol FF,
gives a good colour change from green through grey (end-point) to red, which
obviates the need for a matching buffer solution.
If carbonate is present, it also reacts with the acid but the amount of acid
reacting with the bicarbonate only can be calculated. As in the determination
of carbonate, standard sulphuric acid.
Procedure
For bicarbonates estimation milligrams of carbonates present subtracted
from methyl orange alkalinity.

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Nitrates
Principles
The calorimetric methods generally used for nitrate employ diazotizing
reagents. Nitrates react with these reagents in acidic solution to form
diazonium salts. The diazonium salts are coupled with amino or hydroxyl
groups of aromatic compounds to form coloured azo compounds.

Apparatus
Spectrophotometer
Pipettes
Flasks (100,500,1000ml)
Test tubes
Reagents
Chromotropic acid
H2SO4
HCl
Procedure
Standard curve
For standard curve of nitrate stock solution of 100 ppm nitrate-nitrogen (NO 3
–N) was prepare by dissolving 0.7216 g of pure KNO 3/liter of deionized water.
From this stock solution working solutions of 2, 4, 6, 8, 10, 12, 14, 16, 18, 20
ppm were prepared by dilution. Colour was developed by modified method of
Sims and Jackson (1971) as described by Kowalenko and Lowe (1973). A
brief description of the method is as follows:
Preparation of CTA solution
A 0.1 percent solution of practical grade chromotropic disodium salt
(CTA) in concentrated H2SO4 was prepared by dissolving 0.184 g of CTA in
100 ml of reagent grade H2SO4 (specific gravity =1.84 g/cc). A 0.01 percent
working solution of CTA was prepared by diluting 10 ml of the 0.1 percent

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solution of CTA and 1 ml of concentrated HC1 and 100 ml concentrated
H2SO4.
Aliquots (3 ml) from each of the standard series were taken in test tubes
and 7 ml of 0.01 percent working solution of chromotropic acid was added by
thrust of pipette filler. The dark black CTA-NO 3 colour was developed. Later
on the solution was mixed with a glass rod to homogenize and absorbance
measurement was made at 430 nm with the help of “Spectrophotometer”.
Absorbance was plotted against concentrations to get a linear curve or direct
concentration was measured by Spectrophotometer.
Filter a water sample of 10ml through whatman # 42 or equivalent filter
paper. Take 3 ml from each of the water samples in test tubes and 7 ml of 0.01
percent working solution of chromotropic acid was added by thrust of pipette
filler. The dark black CTA-NO3 colour was developed. Later on the solution
was mixed with a glass rod to homogenize and absorbance measurement was
made at 430 nm with the help of “Spectrophotometer” and see the value from
the graph or direct concentration was measured by Spectrophotometer.

Phosphates (soluble orthophosphate)

Principles
In an acid solution, orthophosphate react with ammonium molybedate to
form an ammonium phosphomolybdate complex. The molybdenum in the
complex can be reduced to blue-coloured solution. The intensity of the blue
coloured from this solution increases in proportion to the amount of phosphate
present since uncomplexed molybdenum in solution does not react with the
reducing agent. Stannous chloride has been widely used for this purpose in
water analysis.

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Apparatus
Spectrophotometer
Pipettes
Burettes
Reagents
Ammonium molybedate (dissolved 25g of Ammonium molybedate in
175ml of distilled water add 280ml of conc. H 2SO4 make a volume of 1000ml
in distilled water)
Stannous Chloride (dissolved 2.5g of fresh Stannous Chloride in 100ml
of glycerol by heating in water bath and stirring with a glass rod)
Standard Phosphate-Phosphorus solutions (dissolved 0.2195g of
KH2PO4 in distilled water and make a volume of 100ml in volumetric flask and
make required dilution from this stock solution)
Procedure
Phosphate estimation in water was made by the following method as
described in A.P.H.A. (1975). This was modified as described bellows:
Instead of standard curve a standard factor was calculated and then from
this unknown concentration of water samples were estimated. A standard stock
solution of phosphate was prepare by dissolving 0.1433 g potassium phosphate
dried overnight at 80 oC in one liter of distilled water, this stock was contained
0.10 mg/PO4/ml From this stock the working standards of 0.01, 0.02, 0.03,
0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10 pmm were prepared.
A 5 percent stock solution of ammonium molybedate was prepared by
dissolving 10 g of ammonium molybedate in distilled water and diluted it to
200 ml. Another stock solution of H2SO4 (1+6) was also prepared by
dissolving 1 ml conc. H2SO4 in 5 ml of distilled water.
Aliquots (2ml) from each of the standard series were taken in test tubes
and 2 ml of 5 percent working solution of ammonium molybedate and 2 ml of
H2SO4 (1+6) was added successively. Later on the solution was mixed with a
glass rod to homogenize and allowed to stand for 15 to 30 minutes. Similar

6
treatment was given to the unknown samples. The absorbance of the test
sample and standards against the blank were recorded on Bausch and Lomb
“Spectronic-21” at a wave length of 630 nm. Standard factor calculations of
unknown concentrations were estimated by the following formula:

Concentration
Standard factor (F) = _____________________
Absorbance at 400 nm (A)

Concentration = F x A

TOTAL HARDNESS