110 PHYSICAL AND CHEMICAL METHODS IN SOIL ANALYSIS

Since, platinum is too soft in pure condition, it is alloyed for stiffening, usually with a
fraction of a per cent of irridium. The life of platinum utensils are maintained well by (a) keeping
them chemically clean, (b) avoiding specific reagents in which platinum is soluble or reactive
and (c) handling them so as to keep them in good mechanical condition.
Cleaning Procedure
● Crucibles should be cleaned singly
● Soaking in hot water and scrubbing
● If not clean then boiling in 6(N) HCl for few minutes
● If not clean the crucible is dried and fused with potassium pyrosulphate (K2S2O7), the
melt is poured into dry waste sand and the residue dissolved from the crucible in
warm 6(N) HCl.
● Crucibles can be further cleaned by digestion with little HF to which 3 drops of H2SO4
have been added.
● Crucible is warmed for few minutes in 6(N) HCl, rinsed with distilled water and dried
in an oven.
Specific Reagents in which Platinum is Soluble or Reactive Must be Avoided
● Chlorine attacks platinum. Hence platinum utensils must not be digested in aquaregia
from which chlorine is liberated. Ferric chloride in presence of HCl must not be used.
● Hydroxides, oxides, peroxides, nitrites and cyanides of alkalies strongly attack platinum.
Handling of Platinum Utensils
● The utensils must be handled in such a manner so as to prevent deformation.
● Platinum-tipped or pure nickel tongs are employed in handling the pt-crucible. Brass,
nickel plated or iron tongs should never be used.
● A porcelain plate, beaker or asbestos pad is employed to set the platinum utensil on,
never a desk top or ring stand.

3.13 AMMONIUM ACETATE EXTRACTABLE POTASSIUM

Principle
The readily exchangeable plus water soluble K+ is determined in the neutral normal
ammonium acetate extract of the soil. The NH4+ ion provides a sharp and quick separation from
the exchange sites while other cations bring about a gradual replacement of either more or less
amount of potassium which normally increases with the period of contact. Since, NH4+ holds
highly charged layers together just as K+, the release of non-exchangeable K+ to exchangeable
form is retarded during NH4OAc extraction [Ammonium ions undergoes equilibrium fixation in
the 2 : 1 layer silicates, particularly in the highly charged vermiculite interlayer spaces, in
exactly the same way as K+, by closure of the interlayer space. The ammonium ions thus fixed
undergoes only slow exchange and is reluctant to nitrify, Na+ ions best replaces NH4+ and K+
from slow exchange position].
Non-exchangeable K also has been found to contribute appreciably towards potassium
availability to crops. The commonly used extractant for such purposes involve hot 1(N)HCl and
boiling 1(N)HNO3. The procedure for determination involves either prior removal of exchangeable
K+ or conditions made sufficiently vigorous to extract both exchangeable (including water soluble)
and a portion of non-exchangeable forms from which the former is subtracted.
SOIL CHEMISTRY 111

Reagents
● Neutral normal ammonium acetate solution; Dilute 60 ml glacial acetic acid (99.5%)
and 75 ml concentrated ammonia solution (sp. gr. 0.91, 25% NH3) to one litre. Mix
well, cool and adjust the pH to 7.0 with dilute acetic acid or ammonia solution.
● Potassium chloride solution : 1000 ppm stock solution; Dissolve 1.907 g of AR grade
potassium chloride (dried at 60°C for 1 hr.) in distilled water and make up the volume
to 1 litre.
Procedure
● Weigh 5 g soil sample in a 25 ml conical flask.
● Add 25 ml of neutral normal ammonium acetate (pH = 7) and shake for 25 minutes.
● Filter immediately through a dry filter paper (Whatman No.1).
● Reject first few ml of the filtrate.
● Determine the potassium concentration in the extract flamephotometrically after
necessary setting and calibration of the instrument.
Standard curve for potassium
● From the mother stock solution (1000 ppm K), prepare 2, 5, 10, 15 and 20 ppm K
solutions in 50 ml volumetric flask by proper dilution.
● Adjust the gas and air pressures of the flamephotometer (as per direction given in the
operation manual) and set to the appropriate filter.
● Adjust the flamephotometer reading to zero with the blank (ppm) and at 100 for the
maximum ppm, say 20 ppm.
● Construct the standard curve by plotting the flamephotometer readings along Y-axis
and the different concentrations (ppm) along X-axis.
● Draw a mean line passing through the origin i.e. (0,0) coordinate.
● Find out the concentration of the unknown sample by fitting in the standard curve.
Calculations
volume of extract
Available K+ (ppm) = R ×
weight of soil taken
where R = ppm of K+ in the extract, obtained from the standard curve.
Basis of Calculation
Let ‘x’ ppm is the concentration of the test solution obtained from the standard curve.
Suppose ‘d’ times dilution was made (if the original extract is too much high in K+).
Then extractant concentration = (x × d) ppm
Now 106 ml solution contains (x × d) g of K+
LM (x × d) × 25OP g of K
∴ 25 ml solution contains
N 10 Q
+
6

Again 5 g soil sample contains M

L (x × d) × 25OP g of K
N 10 Q
+
6

∴ 10 g soil sample contains M

L ( x × d) × 25 × 10 OP g of K = L(x × d) × 25 O ppm of K
6
6

N 10 6
5 Q
+
MN 5 PQ
+
112 PHYSICAL AND CHEMICAL METHODS IN SOIL ANALYSIS

3.14 CATION EXCHANGE CAPACITY (CEC)

Principle
When a sample of soil is placed in a solution of a salt, such as ammonium acetate,
ammonium ions are adsorbed by the soil and an equivalent amount of cations is displaced from
the soil into the solution. This reaction is termed as ‘cation exchange’, and the cations displaced
from the soil are referred to as ‘exchangeable’. The surface-active constituents of soils that have
cation-exchange properties are collectively termed as ‘exchange complex’ and consists for the
most part of various clay minerals and organic matter. Soil mineral and organic colloidal particles
have negative valence charges that holds dissociable cations and are thus called ‘colloidal
electrolytes. The cation exchange capacity determination involves measuring the total quantity
of negative charges per unit weight of the material. Stated otherwise the total amount of
exchangeable cations that a soil can retain is designated as the cation exchange capacity and is
usually expressed as milliequivalents per 100 g soil or [cmol(p+)kg–1]. The determination of
CEC is of fundamental importance in soil chemistry research. Adsorption, desorption and
leaching of fertilizers, thermodynamic study of ion exchange; retention and release of nutrients,
agrochemicals, soil pollutants, all depends upon the exchange capacity of the soil; CEC is also
found to be an important parameter for soil classification.
The cation exchange capacity is usually measured by leaching the soil or colloid with
neutral normal ammonium acetate. Then the excess salt is removed by washing with 95% ethanol.
The ammonium ion (NH4+) is then determined by steam distillation with magnesium oxide in
an alkaline medium. The ammonia evolved is adsorbed into a known quantity of the standard
acid containing methyl red indicator and the excess acid back titrated with a standard alkali.
Reagents
● 1(N) NH4OAc adjusted to pH = 7; Dilute 60 ml glacial acetic acid (99.5%) and 75 ml
concentrated ammonia solution (sp.gr.0.91, 25% NH3) to 1 litre. Mix well, cool and
adjust the pH of the solution to 7.0 with dilute acetic acid or ammonia solution.
Alternatively, weigh 77.08 g NH4OAc and dissolve in one litre distilled water and
adjust the pH to 7 carefully with dilute acetic acid or ammonia solution.
● Ethanol 60%
● Ammonium chloride (AR)
● Magnesium oxide-carbonate free, freshly ignited (ignite at 650°C for 2 hours and cool
in a desiccator over KOH pellets, store in tightly stoppered bottle)
● Standard H2SO4 ; 0.1 (N)
● Standard NaOH ; 0.1 (N)
● Standard oxalic acid – 0.1 (N)
● Methyl red indicator
● NaOH; 45%
● Silver nitrate solution about 0.1 (M) : Dissolve 8.5 g AgNO3 in 500 ml water. Add 2 ml
concentrated HNO3 and mix well.
Procedure
● Transfer without loss 10 g of air dry soil sample accurately weighed in a 250 ml beaker
and add 50 ml of neutral normal ammonium acetate solution.
● Stir occasionally for an hour cover with watch glass and leave overnight.
SOIL CHEMISTRY 113

● Filter the contents through Whatman No. 44 filter paper receiving the filtrate in a 250
ml volumetric flask.
● Transfer the soil completely on to the filter paper and continue to leach the soil with
1(N) NH4OAc (using 20 ml at a time), allowing the leachate to drain out completely
before adding a fresh aliquot.
● Continue the process, until the flask is full to the mark.
● Preserve this for estimation of exchangeable bases (Na+, K+, Ca++ and Mg++). The recidue
left on the filter paper is intended for determination of cation exchange capacity of the
soils.
● Wash the recidue left on the filter paper with 60% alcohol to remove excess ammonium
acetate. To ensure this add a pinch of solid NH4Cl to the recidue on the filter paper
and wash with alcohol till the filtrate is free from chloride (as tested with silver nitrate
solution, the filtrate is perfectly clear when free from chloride). If the washing is to be
interrupted such as for the night, attach a rubber tube to the tail of the funnel and
pinch it tight with a clip when there is solution above the level of soil in the filter
paper. i.e. in no case the soil should dry otherwise loss of ammonia may occur.
● Remove the soil with the filter paper into a 800 ml distillation flask and add about 200
ml of water and about 3 g MgO (one spoonful approximately).
● Add few glass beads and little liquid paraffin so as to avoid bumping and frothing
during distillation.
● Pour 100 ml of 45% sodium hydroxide and immediately connect the distillation flask
to the condenser and distill ammonia in a known excess of 0.1(N) H2SO4 (say 25 ml) to
which a few drops of methyl red indicator is added. (Continue distillation to collect
about 150 ml distillate).
● Back titrate the excess of acid with 0.1(N) NaOH. Standardize NaOH versus oxalic
acid and H2SO4 versus standard NaOH. [see standardization technique, article no. 3.7].
● Perform a blank distillation without the soil on a similar volume of liquid.
Calculations
CEC is normally expressed in milliequivalents of the cation per 100 g soil, presently as
c mol/(pt) kg–1. Milliequivalent means the equivalent weight expressed in milligrams. For
instance 20 g of Ca2+ represents 1 equivalent or 1000 milliequavalent Ca2+. Likewise 18 mg
NH4+ would represent 1 milliequivalent (meq) of NH4+.
Since 1000 ml of 1(N) acid or alkali = 1.0 g equivalent of any cation.
It follows that 1000 ml of 1(N) acid or alkali = 1000 milliequivalents of any cation.
Therefore, 1 ml 1(N) acid or alkali = 1 milliequivalents of any cation.
LM 100 OP
N
Cation exchange capacity = (V1N 1 − V2 N 2 ) ×
w Q
c mol (pt) kg–1
where V1 = ml of standard acid taken initially for ammonia absorption
N1 = normality of standard acid
V2 = ml of standard base used in back titrating of excess acid
N2 = normality of standard base
w = weight of sample in g.
114 PHYSICAL AND CHEMICAL METHODS IN SOIL ANALYSIS

Fundamental Basis of Calculation

From standardisation of NaOH versus H2SO4
LM N OP H SO
25 ml of
N 10 Q2 4 X ml Y(N) NaOH

[For convenience in calculation carry out standardisation pipetting 25 ml H2SO4]

∴ For back titration process
LM N OP H SO
25 ml of
N 10 Q2 4 Z ml Y(N) NaOH.

Hence, equivalent Y(N) NaOH consumed by NH4+ ion is (X-Z) ml.

We know 1 ml 1(N) NaOH = 1 meq.
∴ (X – Z) ml Y(N) NaOH = [1 × (X-Z) × Y] meq.
Again w g sample contains [(X-Z)Y] meq.
LM 100OP
N
∴ 100 g sample will contain (X - Z)Y ×
w Q
meq.

where,
X is the volume of standard NaOH required for standardisation of 25 ml (N/10) H2SO4.
Z is the volume of standard NaOH required during back titration
Y is the normality of NaOH.
Note
● The exchangeable cation analysis of saline and alkali soils is subject to difficulties not
ordinarily encountered with other soils. Saline and alkali soils commonly contain
alkaline-earth carbonates and a relatively high concentration of soluble salts. They
may also have low permeability to aqueous solution and to alcohol. The method
described above is for non-calcareous soils. In soils containing considerable calcium
carbonate, saturation with ammonium will be only partial so long as the carbonate is
present because of its solubility in the NH4OAc solution. The soluble salts should not
be washed out of the soils prior to extracting the exchangeable cations, because of
significant changes that take place as a result of dilution and hydrolysis. The dissolving
of salt therefore necessiates independent determinations of soluble cation contents
and correction of the exchangeable cation analysis for their presence, while the
occurrence of calcium and magnesium carbonates prevents accurate determination of
exchangeable calcium and magnesium. Also, the low permeability of many alkali soils
renders the conventional leaching techniques time consuming and inconvenient.
Although neutral normal ammonium acetate is the salt solution most commonly used
for the extraction of exchangeable cations, some saline and alkali soils fix appreciable
amounts of ammonium and potassium under moist condition. This fixation does not
interfere with the extraction of exchangeable cations but values obtained for cation
exchange capacity are low by amounts equal to the quantity of ammonium fixed. Thus
using a cation not subject to fixation for CEC determination is necessary for such soils.
NH4OAc method may also give low result if the soil contain predominantly 1 : 1 type
clay minerals (kaolinitic) or much organic matter. Usually, the CEC is determined by
measuring the milliequivalents of sodium adsorbed per 100 g of soil upon treating a
soil sample with an excess of normal sodium acetate solution adjusted at pH 8.2. The
SOIL CHEMISTRY 115

fact that sodium is a prominent cation in most saline and alkali soils also favours its
use in the determination of CEC.
● The method of determination of cation exchange capacity must be reported with the
result as because the use of different saturating cations may lead to different results
due to variation in cationic size, hydration and electric charge affecting the mechanism of
exchange. Cation exchange capacity is not necessarily an absolute constant for a particular
soil but may have a range of values according to the cation involved in its determination.
● The direct distillation of soil in an alkaline medium may lead to partial breakdown of
organic matter thus introducing a possible error with most surface soils. Also if cation
exchange capacity is large, the final titration will consume too much titrant if 10 g soil
is taken. Hence for direct distillation of heavy clay soils mainly montmorillonite, it is
better to take 5 g soil for analysis.
● The object of using neutral alcohol is to remove excess of occluded ammonium acetate
since the ammonium complex undergoes slight hydrolysis if water is the leaching agent.
Further the ammonium saturated soil is highly dispersed when in contact with water
and the fine particles of the soil show a tendency to pass through the filter paper.
● An alternative and better method is to collect the distillate (ammonia) into a 250 ml
conical flask, containing known excess (50 ml) of 2% boric acid solution with mixed
indicator (bromocresol green and methyl red) and is titrated with standard sulphuric
acid (N/10).
3.14.1 Cation Exchange Capacity of Soils Containing Calcium Carbonate
In order to determine the exchange capacity when calcium carbonate is present in soils,
recource should be had to a reagent in which the calcium carbonate is insoluble, and which
contains a cation which is easily analysed after it replaces the exchangeable calcium. The reagent
commonly used is 1(N) sodium acetate adjusted to pH 8.2. When the soil is thoroughly mixed
with this reagent, the exchangeable cations are replaced by sodium. Excess of sodium acetate is
removed by washing with 95% ethanol. This removal is the critical step in the procedure since
exchangeable sodium is easily hydrolysed leading to under estimation of the exchange capacity
of soils and therefore removal of salt is monitored via electrical conductivity measurement of
the washings. Thereafter, the sodium on the exchange sites are displaced by leaching with 1(N)
magnesium nitrate solution adjusted to pH 8.6 and sodium determined through flame
photometry.
Reactions
CH3 COONa CH3COO– + Na+
Soil X + Na+ + CH3COO– Soil – Na + CH3COOX
Mg(NO3)2 Mg2+ + 2NO3–
Soil – Na + Mg2+ Soil – Mg + 2Na+
Reagents
● Sodium acetate solution 1.0 (N), pH 8.2.
Dissolve 82 g anhydrous sodium acetate or 136 g. Sodium acetate trihydrate in about
90 ml water. Adjust the pH to 8.2 with dilute NaOH or dilute acetic acid and dilute to
1000 ml.
● Ethanol 95%
● Magnesium nitrate 1.0 (N) pH 8.6
116 PHYSICAL AND CHEMICAL METHODS IN SOIL ANALYSIS

Procedure
● Weigh 5 g soil sample and place in centrifuge tube.
● Add 33 ml NaOAc, Stopper the tube and shake for 5 minutes.
● Centrifuge the tubes for 10 minutes at about 8000 rpm.
● Decant the supernatant liquid as completely as possible and discard. Repeat this step
four times.
● Add about 30 ml ethanol to each tube, stopper and shake for 5 minutes.
● Centrifuge until the supernatant liquid is clear.
● Decant and discard the supernatant liquid.
● Continue washing until the electrical conductivity of the supernatant liquid from the
last washing is between 40 and 55 µmhos cm–1 (Check the conductivity of each washing
starting from the third. Usually four-five washings are sufficient).
● Replace the adsorbed sodium from the sample by extracting with three 30 ml portions
of Mg(NO3)2 solution.
● Dilute to 100 ml and determine the sodium concentration flamephotometrically or by
Atomic Absorption Spectrophotometer.
Calculations
v1 100
CEC of soil [cmol(p+)kg–1] = c × ×
1000 w
where c is the Na content of Mg (NO3)2 extract (meq/l)
v1 = Volume of extract (ml)
w = Weight of soil sample (g)

3.15 ANION EXCHANGE CAPACITY (AEC)

Anion exchange capacity (AEC) is defined as the quantity of phosphate bound at pH 4 or
5.7. Many anions are often involved in anion exchange reactions viz. PO4=, SO4=, NO3–, Cl– etc.
However, phosphate is usually very suitable for AEC estimation. Under low pH and high
concentration, anions may be adsorbed and exchanged on soil colloids. The adsorption usually
occurs on surfaces having a positive charge, viz. iron and aluminium hydroxides. The anion
retention is related to the nature of anions and that of the soil surface together with amphoteric
properties of organic colloids as well as iron and aluminium hydroxides. In highly acidic soil
conditions phosphorus acid anions are retained directly on the surface of colloidal particles
from the soil solution by adsorption phenomena. The mechanism of anion exchange may be
illustrated as follows:
· By the addition of a proton (H+ ion) to the –OH group linked to a sesquioxide clay
particle (R) i.e. Al2O3, Fe2O3, etc.
R – OH + HOH → R – OH2OH
R – OH + HCl → R – OH2Cl
● By the addition of a proton to the functional groups of the organic fraction in acid soil

R – COOH + H+ → R – COOH2+ + Cl– → RCOOH2Cl

The study of AEC of soils helps in understanding the retention and release mechanism of
important plant nutrient anions, viz. sulphate, phosphate, nitrate particularly in light textured
soils of humid tropics.
SOIL CHEMISTRY 117

Principle
The method involves initial leaching of the soil with a solution of barium chloride-
triethanolamine buffered at pH 8.1, followed by calcium saturation. The Ca-saturated soil is
equilibrated with standard phosphoric acid solution and the quantity of phosphorus adsorbed is
evaluated. From this adsorbed phosphorus plus phosphorus extracted initially the AEC of the
soil is calculated using the formula.
AEC (meq./100 g soil) = [(extractable P + adsorbed P)] expressed as meq./100 g soil.
Reagents
● Calcium chloride solution; Dissolve 50 g CaCl2.2H2O in 100 ml of distilled water and
adjust to pH = 8.0 with saturated Ca(OH)2 solution.
● Triethanolamine solution; Dilute 90ml of triethanolamine to 100ml and adjust the pH
to 8.1 with HCl. Dilute to 200ml and mix equal volume of distilled water containing
100g of BaCl2.2H2O.
● Phosphoric acid solution [0.01 (M) in H3PO4]
● Bray’s (I) Reagent for P extraction [0.025(N) NH4F in 0.03 (N) HCl].
● Dikman and Bray’s reagent for colour development
KH2PO4. stock solution of P for standard curve construction. (see article no.3.11).
● Ethanol – 95%
Procedure
● Weigh 10 g soil and leach with 100 ml of triethanolamine and wash 6 times with 95%
ethanol.
● Leach the soil with 100 ml of CaCl2 solution and wash again.
● Dry the calcium saturated soil at 45°C and weigh into a centrifuge tube sufficient to
give a CEC of about 0.2 meq.
● Add 20ml phosphoric acid solution and shake for half an hour and let stand for 24
hours. Again shake for half an hour. Centrifuge and take 1 ml aliquot for P-estimation.
● In a separate soil sample, extract ‘P’ with Bray’s reagent and determine ‘P’ colorime-
trically using chloromolybdic acid reagent.
Calculations
Weight of soil taken = 10 g
Volume of phosphoric acid solution added = 20 ml.
Volume of aliquot taken = 1 ml.
Let this 1 ml is made upto V ml. and concentration of P from standard curve = C ppm.
20
Hence first dilution = = 2 times
10
V
Second dilution = = V times
1
Total dilution = 2V times.
Therefore, concentration of P in solution phase = (2 × C × V) ppm.
FG X IJ meq./100g
Thus P adsorbed = [P added (ppm) – 2 C.V] = X ppm =
H 6.2 × 10 K
118 PHYSICAL AND CHEMICAL METHODS IN SOIL ANALYSIS

FG Y IJ
Also, extractable P = Y ppm =
H 6.2 × 10 K
L ( X + Y ) OP
AEC (meq/100g soil) = M
So,
N 62 Q
3.16 EXCHANGEABLE BASES
3.16.1 Exchangeable Sodium
Principle
Sodium is readily excited in a flame producing an intense yellow light, the yellow colour
is primarily due to radiation of 589.6 millimicron wavelength popularly known as D-line of
sodium. Other less powerful radiations of different wavelength emitted are effectively blocked
by a suitable yellow glass (Na-filter) allowing only the D-line emission to pass through. Thus, if
a solution containing sodium ions is fed as a fine spray into a flame under controlled and standard
instrumental conditions and the emitted light is passed through a Na-filter, the intensity of the
D-line emission can be easily measured photoelectrically and related to the concentration of the
sodium in the test solution. The flamephotometer is calibrated with a series of standard sodium
chloride solution and then used to determine the unknown sodium concentration of the solution
under analysis within the same range.
Procedure
● Analyze directly the ammonium acetate extract for Na+ and K+ in the flamephotometer.
Standard Curve for Sodium
● Dissolve accurately weighed 2.542 g NaCl in distilled water and make up the volume
to one litre. This gives 1000 ppm stock solution of Na+.
● From this prepare 1, 2, 3, 4, 5, 6, 7 and 10 ppm Na+ by proper dilution.
● Adjust the gas and air pressures of the flamephotometer as per direction given in
operation manual and set to appropriate filter.
● Adjust the flamephotometer reading to zero with blank (0 ppm) and 100 for the
maximum (10 ppm).
● Construct the standard curve by plotting the flamephotometer reading along x-axis
and concentrations along y-axis.
● Draw a mean line passing through the origin.
● From this graph obtain the sodium concentration of the sample under analysis in
milliequivalents per litre or in ppm. If there is a dilution of the original sample, multiply
by the dilution factor.
● Check the performance of the flamephotometer at frequent intervals by spraying some
standard solutions and adjusting the sensitivity as necessary.
Calculations
volume of extractant
Exchangeable Na+ (ppm) = R ×
weight of soil
where R = ppm of Na in the extract as obtained from the standard curve. R must include any
dilution factor, if used.
also, 1 meq./l Na = 23 ppm Na.
SOIL CHEMISTRY 119

Note
● Standard solutions for Na are also prepared in meq./l rather than ppm. For this a
stock solution of 0.05 (N) NaCl is prepared by dissolving accurately weighed 1.4625 g
of dry NaCl in 500 ml distilled water. From this stock 2,4,6,8 and 10 ml solution is
diluted to one litre respectively to get working standards containing 0.1, 0.2, 0.3, 0.4
and 0.5 milliequivalents per litre.
● In analysis of water or soil extracts, the only ion which may cause serious interference

to sodium measurements is calcium. This usually happens when calcium occurs in a

much higher concentration than sodium. For instance, water extracts of gypsiferrous
soils can contain up to 30-32 meq/l of calcium while sodium level may be less than 1
meq/l. In general for a particular instrument and type of flame, there may be
interference effects from some of the other cations or anions present in the test solution
and these effects must either be suppressed or measured. The calcium ion sometimes
tend to enhance sodium emission and the effect may be measured for a range of calcium :
sodium ratios and appropriate corrections applied or in some cases the interference
may be suppressed by addition of aluminium nitrates.
Usually, a series of standard NaCl solution is prepared containing 0.1–0.5 meq/l Na+ by
dilution from 0.05(N) NaCl solution using saturated calcium sulphate solution in place of water.
Since mostly the calcium in soil extract is associated with sulphate these solutions contains
about 30 meq/l calcium as sulphate. Next to this, the flamephotometer is calibrated with pure
NaCl standard and subsequently the standard containing the calcium sulphate is sprayed and
any interference effect due to calcium is measured.
However, sodium may be determined also by atomic absorption spectrophotometry using
emission mode. Calcium does not normally interfere in this technique.

3.16.2 Exchangeable Calcium and Magnesium

Where atomic absorption spectrophotometry is possible the ammonium acetate extract
can be directly analysed for Ca and Mg. The spectrophotometric standards are prepared in the
ammonium acetate solution and both the standard and extracts are read against ammonium
acetate as blank. If AAS is not possible the calcium and magnesium are analysed by
complexometric titrations using ethylene diamine tetra acetic acid (EDTA).
Principle
The method makes use of excellent chelating properties of disodium ethylene diamine
tetraacetate (versenate) which forms soluble complexes with metal cations.
The structure of EDTA may be represented as follows :
120 PHYSICAL AND CHEMICAL METHODS IN SOIL ANALYSIS

The formula II is preferred over I, since it has been shown from measurements of the
dissociation constants that two hydrogen atoms are probably held in the form of zwitter ions.
For the purpose of simplicity we shall assign the formula H4Y to EDTA : the di-sodium salt is
therefore Na2H2Y and affords the complex-forming ion H2Y2 in aqueous solution; it reacts with
all metals in a 1 : 1 ratio. The reactions with cations M2+, can be written in the general form as :
M2+ + H2Y2– = MY2– + 2H+
M3+ + H2Y2– = MY + 2H+
or Mn+ + H2Y2– = (MY)(n–4)+ + 2H+
One mole of complex forming H2Y2– reacts with one mole of the metal ion and in each
case two moles of hydrogen are formed. It is evident from the equations above that dissociation
of the complex will be governed by the pH of the solution; lowering the pH will decrease the
stability of metal-EDTA complex. The more stable the complex, the lower the pH at which an
EDTA titration of the metal ion in question is carried out.
The stability of a complex is characterised by the stability constant (or formation con-
stant) K:
Mn+ + Y4– = (MY)(n–4)+
[(MY ) ( n − 4) + ]
∴ K=
[M n + ][ Y 4 − ]
Assuming the fully ionised form of EDTA i.e. the ion Y4– has been taken into account, but
at two pH-values the species HY3–, H2Y2–, H3Y– and even undissociated H4Y may be present;
stated otherwise only a part of the EDTA uncombined with metal may be presented as Y4–.
Further, the metal Mn+ is assumed to be uncomplexed i.e. in aqueous solution, it is simply
present as the hydrated ion.
The success of an EDTA-titration depends upon the precise determination of the end
point. The most common technique is to use metal-ion indicators. The requisites of a metal ion
indicator for use in the visual detection of end point include :
● The colour reaction must be such that before the end-point when nearly all the metal

ion is complexed with EDTA, the solution is strongly coloured.

● The colour reaction should be specific.

● The metal-indicator-complex must posses sufficient stability, otherwise because of

dissociation, a sharp colour change is not obtained. The metal indicator complex
however, must be less stable than the metal-EDTA complex to ensure that, at the end
point, EDTA removes metal ions from the metal-indicator complex.
● The colour contrast between the free indicator and the metal-indicator complex should

be such as to be readily observed.

The use of a metal ion indicator in an EDTA titration may be written as :
M-In + EDTA → M-EDTA + In.
The reaction will proceed if the metal-indicator complex (M-In) is less stable than the
metal-EDTA complex (M-EDTA). The former dissociates to a limited extent and during the
titration the free metal ions are progressively complexed by the EDTA until ultimately the
metal is displaced from the complex (M-In) to leave the free indicator (In).
Some of the metal ion indicators used for calcium and magnesium are discussed below;
Solochrome dark blue or calcon
This is sodium 1 – (2-hydroxy-1-napthylazo)-2-napthol-4-sulphonate
SOIL CHEMISTRY 121

An important application of this indicator is in the complexometric titration of calcium in

presence of magnesium. This must be carried out at a pH of about 12.3 in order to avoid inter-
ference of magnesium (obtained with a diethylamine buffer; 5cc/100cc solution). Under such
condition magnesium is precipitated quantitatively as magnesium hydroxide. The colour change
at end point is from pink to pure blue.
Patton and Reeder’s indicator
The indicator is 2-hydroxy-1-(2-hydroxy-4-sulpho-1-napthylazo)-3-napthoic acid commonly
abbreviated as HHSNNA. Its main application is direct titration of calcium, particularly in
presence of magnesium. A sharp colour change is from wine red to pure blue is obtained when
calcium ions are titrated with EDTA at pH values between 12-14.
Murexide
This is ammonium salt of purpuric acid and its anion has the following structure.
Murexide is of interest because it was probably the first metal ion indicator to be employed
in the EDTA titration. The murexide may be employed for direct EDTA titration of calcium at
pH = 11; the colour change at end point is from red to blue violet.
Fundamental Concept of EDTA Titration
When calcium ions are titrated with EDTA a relatively stable calcium complex is formed
Ca2+ + H2Y2– → CaY2– + 2H+
With magnesium ions, a somewhat less stable complex is formed
Mg2+ + H2Y2– → MgY2– + 2H+
The magnesium-indicator complex is more stable than the calcium-indicator complex
but less stable than Magnesium-EDTA complex.
i.e. [Mg-EDTA] < [Mg-In] > [Ca-In].
Consequently, during titration of solution containing magnesium and calcium ions with
EDTA in presence of Eriochrome Black T the EDTA first reacts with the free calcium ions, then
with the free magnesium ions, and finally with the magnesium-indicator complex. Since mag-
nesium-indicator complex is wine red in colour and the free indicator is blue between pH = 7-11,
the colour of the solution changes from wine red to blue at the end-point .
[Mg-EBT-] + H2Y2– = MgY2– + HEBT2– + H+
(red) (blue)
The titration with EDTA, using Solochrome Black (Eriochrome Black T) as indicator
gives the total calcium plus magnesium content. To determine the individual elements, calcium
may be estimated by titration using Patton and Reeder’s indicator. The difference between the
two gives the estimate of magnesium.
Traces of many metals interfere in the determination of calcium and magnesium using
Eriochrome Black T indicator e.g. Co, Ni, Cu, Zn, Hg and Mn. The interference can be overcome
by addition of a little hydroxylamine hydrochloride which reduces some of the metals to their
lower valency states and also of sodium or potassium cyanide complexes. Iron may also be
rendered harmless by addition of a little sodium sulphide.
Reagents
● Hydrochloric acid (Analytical grade)
● Nitric acid (Analytical grade)
122 PHYSICAL AND CHEMICAL METHODS IN SOIL ANALYSIS

● Standard Ca solution (0.0 1N) primary standard; Dissolve 0.2502 g of pure calcium
carbonate (analytical grade, dried at 110°C overnight) with minimum quantity of
concentrated HCl dropwise (10 ml of 3N HCl may be used). Warm the solution to expel
CO2 and then dilute to 500 ml in volumetric flask.
● EDTA solution approx. 0.01 (N); Dissolve 2.0 g disodium dihydrogen ethylenediamine
tetraacetate (Na2 H2 C10H12O8 N2 . 2H2O) and 0.05 g magnesium chloride hexahydrate
in one litre water and standardize against standard calcium solution.
● Sodium hydroxide solution – 10%
● NH4Cl – NH4OH Buffer (pH = 10) ; Dissolve 67.5 g ammonium chloride (AR) in 570 ml
of concentrated ammonia (sp.gr. 0.91) and dilute to one litre.
● Eriochrome Black T indicator; Take 100 ml of ethanol and dissolve 4.5 g hydroxylamine
hydrochloride in it. Now add 0.5 g of the indicator and prepare solution.
● Calcon indicator; Dissolve 0.20 g of the dyestuff (calcon) in 50 cc methanol; (ethanol
may also be used). Prepare fresh solution weekly.
Note : Sodium diethyl dithio carbamate crystals or 2% sodium cyanide solution, (used generally to
eliminate interference arising due to presence of Cu, Zn, Fe, Mn, Sn, if present in appreciable amounts.
However, in irrigation water and water extract of soil interfering ions are negligible and can be neglected).
Procedure
Pretreatment of NH4OAc Extract
● Ammonium acetate may interfere in EDTA–tiltration and is therefore destroyed by
oxidation with a mixture of HCl and HNO3.
● Pipette 100 ml of NH4OAc extract into a 500 ml beaker and evaporate carefuly to
dryness, cool, and add 5 ml concentrated HCl(washing down salts on wall of the beaker),
followed by 1 ml HNO3.
● Cover with a watch glass immediately.
● When vigorous reaction has ceased evaporate the solution to dryness in a fume hood.
Cool and add 1 ml concentrated HCl followed by 20 ml water, stir well and filter the
solution through a Whatman No. 40 filter paper into a 100 ml volumetric flask.
● Rinse the beaker 3-4 times, collecting the rinsings through the filter paper into the
volumetric flask.
● Make up the volume with distilled water.
● Prepare a blank solution by taking 100 ml ammonium acetate solution by the same
procedure.

Calcium
● Pipette 5 ml of the solution (or a suitable aliquot) into a 100 ml conical flask or into a
porcelain dish and dilute it approximately to 25 ml (add 20 ml water, if 5 ml aliquot is
taken).
● Add 1 ml or more of 10% NaOH to raise the pH to 12. (Check the pH with a pH-meter,
if necessary), (2-3 crystal of carbamate may be added if required).
● Add 10-12 drops of calcon indicator stir the solution and titrate with standard EDTA
solution until the colour changes from pink to pure blue.
● To check the end point accurately, perform a blank titration taking 25 ml water instead
of sample solution and adding other reagents in the similar manner.
● Also determine the blank correction by titrating a similar aliquot of blank solution.
SOIL CHEMISTRY 123

Calcium Plus Magnesium

● Pipette 5 ml of the solution or a suitable aliquot (containing not more than 0.1 meq. Ca
+ Mg) into a dish or 100 ml conical flask and dilute to about 25 ml.
● Add 10 ml of NH Cl–NH OH buffer, (2-3 carbamate crystals if required) and 3-4 drops
4 4
of Eriochrome Black T indicator.
● Titrate against standard EDTA solution until colour changes from red to permanent

blue colour.
● Perform a blank by replacing sample with 25 ml distilled water.

● Also determine the blank correction.

Standardise the EDTA solution with standard Ca solution using Eriochrome black-T
indicator or calcon indicator (see note below), using the procedure for calcium-estimation.
Note
● For analysing water samples, NH4OAc-pretreatment is not required and hence ‘blank
correction’ is also not done.
● If the EDTA solution is prepared by dissolving 2.0 g of the salt in one litre water
without addition of 0.05 g magnesium chloride hexahydrate, then use calcon indicator
during standardization of EDTA with standard calcium solution.
Calculations
Exchangeable calcium (meq/100 g soil).
LM V − V
1 2
× V4 × N ×
100 OP
=
N V 3 w Q
where V1 = volume of EDTA required for sample aliquot titration (calcon), ml
V2 = volume of EDTA required for blank titration (calcon), ml
V3 = volume of aliquot, ml
V4 = total volume of original NH4OAc extract, ml
N = normality of EDTA
w = weight of sample in g
Exchangeable (Ca + Mg), [meq./100g soil]
LM V − V
5 6
× V4 × N ×
100 OP
=
N V 7 w Q
where V5 = volume of EDTA (ml) required for sample aliquot titration using EBT
V6 = volume of EDTA (ml) required for blank aliquot titration using EBT
V7 = volume of aliquot taken (ml)
V4 = total volume of original NH4OAc extract (ml)
N = normality of EDTA
w = weight of sample taken in g
Note : 1 ml 0.01 (N) EDTA = 0.2004 mg Ca2+ = 0.1216 mg Mg2+

3.17 EXCHANGEABLE CALCIUM AND MAGNESIUM IN CALCAREOUS SOILS

A KCl solution buffered at pH = 8.3 by triethanol amine is used as an extractant.
124 PHYSICAL AND CHEMICAL METHODS IN SOIL ANALYSIS

Reagents
● KCl (1.0 N) – triethanolamine buffer solution ; Dissolve 74.6 g potasium chloride in
about 500 ml water, add 25 ml triethanolamine (sp.gr.1.12) and stir well. Dilute to
about 850 ml and mix. Adjust the pH to 8.2 with 1.0 (N) HCl. About 85 ml 1(N) HCl
will be required. Dilute to one litre.
Procedure
● Weigh 10 g air dry sample into a 100 ml beaker and add 40 ml KCl-triethanolamine
solution. Stir thoroughly frequently for 20 minutes.
● Filter the suspension through Whatman No.40 filter paper into a 100 ml volumetric
flask. Leach with 20 ml portions of the buffer solution to bring the volume of leachate
to 100 ml.
● The extract can be analyzed for Ca and Mg directly by AAS. Prepare the standards (in
terms of meq/l) in the KCl-triethanolamine solution.
● Read standards and test samples against the buffer solution as blank.
● If AAS is not possible, determine Ca and Mg by EDTA/versenate method as described
previously. (No pretreatment with HCl-HNO3 is required).
● Determine a blank for KCl-triethanolamine solution using Eriochrome black T and
calcon indicators.
Calculations
Exchangeable Ca (meq/100 g soil)
LM V − V
1 2
×N×
V4
× 100
OP
=
N V 3 w Q
where V1 = volume of EDTA for sample titration using calcon (ml)
V2 = volume of EDTA for blank titration using calcon (ml)
V3 = volume of aliquot taken (ml)
V4 = total volume of KCl-TEA extract (ml)
N = normality of EDTA
w = weight of sample in g.
Exchangeable (Ca + Mg) in meq./100 g soil
LM V − V
5 6
×N×
V4
× 100
OP
=
N V 7 w Q
where V5 = volume of EDTA required for sample titration using EBT (ml)
V6 = volume of EDTA required for blank titration using EBT (ml)
V7 = volume of aliquot taken (ml)
V4 = total volume of KCl-TEA extract (ml)
N = normality of EDTA
w = weight of sample in g.
SOIL CHEMISTRY 125

3.18 MICRONUTRIENTS : DTPA EXTRACTABLE Zn2+, Cu2+, Fe2+, Mn2+

(LINDSEY & NORVELL, 1978)
Principle
The micronutrient cations can be estimated in a single extraction with diethylene triamine
pentaacetic acid (DTPA) which has excellent chelating property with the micronutrient elements.
Adequate precaution must be taken against any likely contamination from the reagents and
glass wares in micronutrient assay work. Only double distilled water should be used. Specific
hallow cathode lamps for each elements are used on AAS and requisite standards for instrument
calibration are prepared as per instructions in the operation manual.
Reagents
● DTPA – 0.005 M solution; Weigh 1.967 g of DTPA and 1.470 g CaCl2.2H2O in a beaker.
To this add 20-25 ml of double distilled water and 13.3 ml of Triethanolamine (TEA)
followed by 100 ml of double distilled water. Transfer to one litre volumetric flask with
3-4 washings and make up the volume up to the mark with double distilled water.
Adjust the pH to 7.3 with dilute HCl (1.5).
● TEA (Triethanolamine)
● CaCl2 . 2H2O (AR)
● Dilute HCl (1 : 5).
Procedure
● Weigh accurately 10 g of soil sample in a 100 ml conical flask and add 20 ml of DTPA
extractant, shake for 2 hours.
● Filter the extract through Whatman No.42 filter paper.
● Estimate the micronutrient cations (Zn2+, Cu2+, Fe2+, Mn2+) with the help of atomic
absorption spectrophotometer.
Calculation
Micronutrient elements (ppm) = 2 × concentration (ppm) obtained from AAS
× dilution factor (if any).
Note : The standard atomic conditions for the respective micronutrient elements are to be fol-
lowed from Instruction Manual of the Atomic Absorption Spectrophotometer.

3.19 ARSENIC DETERMINATION BY CONVERSION TO THEIR HYDRIDES AND

ASPIRATION INTO AN AAS
Arsenic is ubiquitous in nature and is found in detectable concentrations in all
environmental matrices. The occurrence of As in the continental crust of Earth is usually given
as 1.5 to 2.0 mg/l. The distribution of arsenic in nature is extremely variable, showing little
correlation with geological formation, climate, or soil. Numerous minerals, rocks, sediments
and soils contain arsenic partly as constituent of sulfide minerals or complex sulfides of metal
cations and partly as a constituent retained by soils and/or sediments in occluded or adsorbed
forms. The latter is manifested primarily by the adsorption or occlusion of As on hydrous Al and
Fe oxides, but these are not necessarily the only source. Arsenic is also adsorbed on clay colloid,
is bound to organic matter and may form slightly water soluble compounds with Al, Fe, Ca and
Mg in the soil matrix. Some of the more common minerals in soils are arsenopyrite (FeAsS),
Orpiment (As2S3) etc.
126 PHYSICAL AND CHEMICAL METHODS IN SOIL ANALYSIS

Arsenic is a labile element and can exist in several forms and oxidation states (– 3, 0,+ 3
and + 5 valence in nature). In strongly reducing environments, elemental As and As(III) can
exist but As(V) is the stable oxidation state in aerobic environments.
In reduced environments such as sediments, the methanogenic bacteria reduces As(V) to
As(III) and methylates it to methyl arsenic acid. Standard for maximum allowable As
concentration in drinking water has been set to be 0.01 mg/l by World Health Organization
(WHO). Acute As poisoning in human beings is characterized by central nervous system effect,
leading to coma and eventually death. Chronic intoxication results in neurological disorders,
muscular weakness, loss of appetite, nausea, and skin disorders such as hyper-pigmentation
and keratosis.
Principle
The system consists of an atomic absorption spectrophotometer and a hydride generator.
Most atomic absorption spectrophotometer manufacturers now offer hydride generators or
accessories that can quickly be attached to a spectrophotometer and are rather simple to operate.
The system involves generating arsine as hydride, transferring the hydride to a quartz cell
mounted in the light beam of the spectrophotometer, decomposing the hydride within the confines
of the cell heated externally by an air-acetylene flame and finally obtaining a measured absorption
signal.
Arseneous acid, the As(III) oxidation state of arsenic are instantaneously converted by
sodium borohydride reagent in acid solution to their volatile hydrides. The hydrides are purged
continuously by argon or nitrogen into an appropriate atomizer of an atomic absorption
spectrophotometer and converted to the gas phase atoms. The sodium borohydride reagent by
rapid generation of the elemental hydrides in an appropriate reaction cell, minimizes dilution
of the hydrides by the carrier gas and provides rapid, sensitive determination of arsenic. At
room temperature and solution pH values of 1 or less, arsenic acid, the As (V) oxidation state of
arsenic, is reduced relatively slowly by sodium borohydride to As(III), which is then instantane-
ously converted to arsine. Determination of total arsenic requires that all inorganic arsenic
compounds be in the As(III) state by reduction of any As(V) to As(III) with sodium/potassium
iodide, after initial conversion of all inorganic and organic arsenic compounds and standards to
As(V) by digestion. Arsine is evolved by reduction with sodium borohydride (NaBH4) from HNO3
– H2SO4 soil digest media. As carrier gas sweeps the arsine directly into a flame-heated quartz
cell mounted in the optical path of a suitably equipped atomic absorption spectrophotometer.
Note : Certain atomic absorption atomizers and hydride reaction cells are available commercially for use
with sodium borohydride reagent. Irrespective of the hydride reaction cell-atomizer system selected, it
must meet the following quality control considerations:
● It must provide a precise and reproducible standard curve between 0.20 µg As/l and a
detection limit between 0.1 and 0.5 µg As/l.
● When carried through the entire procedure, oxidation state couple [As(III) – As(V)]
must cause equal instrumental response.
● Sample digestion must yield 80% or greater recovery of added (dimethyl arsenic acid)
and 90% or greater recovery of added As(III), As(V).
Caution Arsenic and its hydride is toxic, handle with care.
SOIL CHEMISTRY 127

Apparatus and Reagents

● Atomic absorption spectrophotometers; equipped with gas flow meters for argon (or
nitrogen) and hydrogen, As electrode less discharge lamps with power supply, back-
ground correction at measurement wavelengths, and appropriate strip-chart recorder.
● Atomizer; Cylindrical quartz cell, 10 to 20 cm long electrically heated by external
nichrome wire to 800–900°C. or cylindrical quartz cell with internal fuel rich hydrogen-
oxygen (air) flame. The sensitivity of quartz cells deteriorates over several months of
use. Sensitivity sometimes may be restored by treatment with 40% HF.
● Reaction cell for producing As hydride; A commercially available system is acceptable
if it utilizes liquid sodium borohydride reagents.
● Sodium borohydride reagent; Dissolve 8 g sodium borohydride in 200 ml of 0.1 (N)
NaOH. Prepare fresh daily;
● 10% KI; Add 10g of potassium iodide (KI) and dilute to 100 ml in a flask.
Procedure
Digestion for Total As
● Pulverize dry soil in an agate mortar to pass non-metallic 100 mesh screen.
● Transfer 1.0 g of the dry, pulverized soil into the Kjeldahl flask, add three glass beads,
30 ml of concentrated HNO3 and swirl to mix contents.
● Place flask on micro-Kjeldahl digester in a vented hood.
● Predigest by slow heating for 45 minutes taking care not to permit severe foaming or
bumping. It may also be necessary to rotate flask to prevent soil caking.
● Increase temperature to produce steady boiling and continue process until 2-4 ml of
HNO3 remains in flask.
● Remove from the digester and allow to cool.
● Add 5 ml concentrated H2SO4, swirl to mix, return to digester and heat to boiling.
● After fumes cease, remove from the digester and cool.
● Add 25 ml of saturated ammonium oxalate, swirl to mix and return to digester.
● Continue soil digestion until fuming ceases. Soil digest should be light gray to nearly
white on completion of digestion. The entire process takes about 3 hours.

Arsenic Extraction Using 0.5 (M) Sodium Bicarbonate Solution Adjusted to pH 8.5 (Johnston
and Barnard; 1979).
● Take 5 g air dried sample in a conical flask and add 100 ml of 0.5 (M) NaHCO3 (pH 8.5)
solution.
● Mix thoroughly and shake on a reciprocating shaker for 18 hours.
● Centrifuge for 10 minutes at 2000 r.p.m. Filter through Whatman no.42 filter paper.
● This solution is used for arsenic estimation.

Determination of Arsenic with Sodium Borohydride

To 50 ml of the above extract add 5 ml conc. HCl and 5 ml 10% KI and wait for half an
hour. This solution is feed to the AAS coupled with hydride generator together with sodium
borohydride, concentrated HCl and the purger gas. (Ar or N2) which transports the generated
arsine (AsH3) to the quartz cell aligned with the arsenic hollow cathode lamp (Model : GBC
932B).
128 PHYSICAL AND CHEMICAL METHODS IN SOIL ANALYSIS

Standard Curve
Commercially available Arsenic standard solution 1000 mg/l is available (Merck;
Germany). From this standard desired concentrations are prepared using double distilled water.
Calculation
The As concentration (µg g–1) is obtained directly from the standard curve which is
calibrated in the instrument keeping in mind the proper dilution factor.

3.20 FLUORIDE ESTIMATION IS SOIL AND WATER ; SPADNS METHOD

[Sodium 2-(Parasulfo phenylazo)-1, 8-dihydroxy- 3, 6-naphthalene disulfonate] also called
[4, 5 dihydroxy-3-(para-sulfophenylazo)-2, 7-napthalenedisulfonic acid trisodium salt].
Fluorine is common in terrestrial environment and is always present in plants, soils and
phosphatic fertilizers. As a rule of thumb, the F concentrations in these materials are on the
order of 3 × 100, 3 × 102 and 3 × 104 ppm for plants, soils and phosphatic fertilizers, respectively.
Fluorine is a common constituent of rocks and soils. Very common soil minerals, such as biotite,
muscovite, and hornblende, may contain as such as several percent of F and, therefore, would
seem to be the main source of F in soils. It appears, therefore, that the F content of soils is
largely dependent on the mineralogical composition of the soil’s inorganic fraction. Phosphatic
fertilizers, especially the superphosphates, are perhaps the single most important source of F
in agricultural lands.
Fluorine is not an essential plant nutrient but is essential for animals. However, continuous
ingestion by the animals of excessive amounts of F can lead to fluorosis, and sub optimal levels
in the diet, can have an equally damaging effect. Therefore, plant content of F is of interest to
livestock producers.
Fluorine is found in soils as the singly charged fluoride ion, F- or occasionally as a
component of such complex anions as (BF4–), (AlF6)3– and (S2F6)2–.
The problem of high fluoride concentration in groundwater resources has now become
one of the most important health-related geo-environmental issues in India, since it has
considerable impact, on human physiology. Its deficiency (< 0.6 mg/l) causes dental caries and
excess (> 1.5 mg/l) causes skeletal fluorosis, dental flurosis respectively. The weathering and
leaching processes, mainly by moving and percolating water play an important role in the
incidence of fluoride in groundwater. The various factors that govern the release of fluoride into
water by the fluoride bearing minerals are (i) the chemical composition of water, (ii) the presence
and accessibility of fluoride minerals to water, and (iii) the time of contact between the source
mineral and water.
Principle
This is a colorimetric method and colour development is virtually instantaneous and no
waiting is required before measuring fluoride concentration. Colour determinations are made
photometrically, using a spectrophotometer. A curve developed from standards can be used for
determining the fluoride concentration of a sample or the concentration can be calculated on
the basis of a pair of standards. The latter technique makes use of the fact that the relationship
between fluoride concentration and absorbance (within the range of the method) is linear and
thus that two points can define accurately the position of the line.
Apparatus and Reagents
● Spectrophotometer for use at 570 nm. providing a light path of at least 1 cm.
SOIL CHEMISTRY 129

● Standard fluoride solution

(i) Stock fluoride solution : Dissolve 221.0 mg anhydrous sodium fluoride, NaF in distilled
water and dilute to 1000 ml; 1 ml = 100 µg F–.
(ii) Standard fluoride solution : Dilute 100 ml stock fluoride solution to 1000 ml with
distilled water.
1.00 ml = 10.0 µg F–.
● SPADNS solution : Dissolve 958 mg SPADNS, [Sodium 2-(parasul fophenylazo)-1, 8-

dihydroxy-3, 6-naphthalene disulfonate] in distilled water and dilute to 500 ml. This
solution is stable for at least 1 year if protected from direct sunlight.
● Zirconyl–acid reagent : Dissolve 133 mg zirconyl chloride octahydrate ZrOCl .8H O,
2 2
in about 25 ml distilled water.
● Acid zirconyl–SPADNS reagent : Mix equal volumes of SPADNS solution and zirconyl-

acid reagent. The combined reagent is stable for at least 2 years.

● Reference solution : Add 10 ml SPADNS solution to 100 ml distilled water. Dilute 7 ml

conc. HCl to 10 ml and add to the diluted SPADNS solution. The resulting solution,
used for setting the instrument reference point (zero), is stable for at least one year.
Alternately, use a prepared standard of 0 mg F–/l as a reference.
● Sodium arsenite solution : Dissolve 5g of NaAsO and dilute to 1 L with distilled water.
2
Avoid ingestion.
Procedure
Preparation of Standard Curve
Prepare fluoride standards in the range of 0 to 1.40 mg F–/l by diluting appropriate
quantities of standard fluoride solution to 50 ml with distilled water. Pipette 5 ml each of SPADNS
solution and zirconyl-acid reagent or 10 ml mixed acid-zirconyl SPADNS reagent, to each
standard and mix well. Set spectrophotometer to zero absorbance with the reference solution
and obtain absorbance readings of the standards. Plot the curve of the mg fluoride-absorbance
relationship. Prepare a new standard curve whenever a fresh reagent is prepared. As a alternative
to using a reference, set photometer at some convenient point (0.300 or 0.500 absorbance) with
the prepared zero mg F–/l standard.
Colour Development
Use a 50 ml sample or a portion diluted to 50 ml with distilled water. Adjust sample
temperature to that used for the standard curve. Add 5 ml each of SPADNS solution and zirconyl-
acid reagent or 10 ml acid-zirconyl-SPADNS reagent. Mix well and read absorbance first setting
the reference point of the photometer as above. If the absorbance falls below the range of standard
curve, repeat using a diluted sample.
Calculations
A B
mg F–/l = ×
ml sample C
where A = µg F– determined from plotted curve.
The ratio (B/C) applies only when a sample is diluted to a volume B, and a portion C is
taken from it for colour development,when the prepared 0 mg F–/l standard is used to set the
photometer.
130 PHYSICAL AND CHEMICAL METHODS IN SOIL ANALYSIS

Alternatively calculate fluoride concentration as follows :

X−Y
mg F–/l =
X−Z
where X = absorbance of the prepared 0 mg F–/l standard
Y = absorbance of the prepared sample
Z = absorbance of the prepared 1.0 mg F–/l standard

3.21 DETERMINATION OF LIME REQUIREMENT OF SOIL

(Schoemaker et al. 1961)
For satisfactory plant growth, the soil should have a pH between 6.5 to 7.5, though cer-
tain plants can grow well at low pH like tea and also at high pH like sugarbeet. In India acid
soils are located mostly in eastern, southern and south central parts. Also some soils at higher
elevations in north India are acidic. For sustained agricultural production and higher yields,
through efficient soil management practices, it is essential to lime and acid soil, as it has consid-
erable influence on soil environment, besides correcting soil acidity.
Principle
In this method the soil is equilibrated with a pH 7.5 buffer solution, whereby the reserve
H+ is brought into solution, which results in the depression of pH of the buffer solution, a note
of which is made and interpreted in terms of lime required to raise the pH to a desired value.
Reagents
● Extractant buffer; Dissolve 1.8 g paranitrophenol, 3 g potassium chromate, 2 g calcium
acetate, 53.1 g calcium chloride dihydrate (CaCl2.2H2O) and 2.5 ml triethanolamine in
1 litre of distilled water. Adjust the pH to 7.5 with NaOH.
Procedure
● Determine the pH of the soil sample in 1 : 2.5 soil:water ratio.
● For this weigh 10 g soil and add 25 ml distilled water.
● Shake intermittently for half an hour and record the soil pH. If the pH exceeds 6.0
then this method is not applicable.
● If the measured pH is 6.0 or low then proceed as follows:
● Weigh 5 g soil in a 50 ml beaker.
● Add to it 5 ml of distilled water and 10 ml buffer solution.
● Stir continuously for 10 minutes or intermittently for 20 minutes.
● Determine the soil pH with the pH meter.
● Lime requirement is determined on the basis of soil-buffer pH ready reckoner given
below.
The values in this table are given in tons of pure CaCO3 per acre required to bring the
soil to the indicated pH and thus are required to be converted to their equivalents in the form of
agricultural lime to be used. The figures are multiplied by a factor of 2.43 to express in tons per
hectre.
SOIL CHEMISTRY 131

pH of soil-buffer Lime required to bring the soil to indicated pH

suspension (tons/acre of pure CaCO3)
pH 6.0 pH 6.4 pH 6.8
6.7 1.0 1.2 1.4
6.6 1.4 1.7 1.9
6.5 1.8 2.2 2.5
6.4 2.3 2.7 3.1
6.3 2.7 3.2 3.7
6.2 3.1 3.7 4.2
6.1 3.5 4.2 4.8
6.0 3.9 4.7 5.4
5.9 4.4 5.2 6.0
5.8 4.8 5.7 6.5
5.7 5.2 6.2 7.0
5.6 5.6 6.7 7.7
5.5 6.0 7.2 8.3
5.4 6.5 7.7 8.9
5.3 6.9 8.2 9.4
5.2 7.4 8.6 10.0
5.1 7.8 9.1 10.6
5.0 8.2 10.1 11.2
4.9 8.6 10.6 11.8
4.8 9.1 12.4

3.22 DETERMINATION OF GYPSUM REQUIREMENT OF SOIL

Principle
Presence of large amount of sodium as high as 15% or more in the exchange complex
results in high soil pH (> 8.0) for sodic (alkali) and saline-sodic soils which causes nutritional
imbalances, depletion of soil organic matter, deterioration of soil physical health and also affects
the soil biotic community Gypsum (CaSO4 . 2H2O) is commonly used for soil amendment under
such situation. A fixed weight of soil is equilibrated with a known amount of Ca solution, and
the amount of Ca left in solution is determined by EDTA-titration. The difference between the
amount of Ca added and Ca left in solution, gives the amount of Ca exchanged. Practically it
has been observed that gypsum addition of about 1/3 of the value obtained by this method is
satisfactory in most cases.
Reagents
● Ammonium chloride–Ammonium hydroxide buffer ; Dissolve 67.5 g of NH4Cl in 570
ml of NH4OH (sp.gr.0.86), and dilute to 1 litre.
● Saturated CaSO4 solution; Shake about 5 g CaSO4 . 2H2O with 1 litre of distilled water
for 15 minutes on a mechanical shaker and filter.
● Eriochrome Black T (EBT) indicator; Dissolve 0.5 g of EBT and 4.5 g of hydroxylamine
hydrochloride in 100 ml of 95% ethanol.
132 PHYSICAL AND CHEMICAL METHODS IN SOIL ANALYSIS

● Standard EDTA solution 0.01N; Dissolve 2 g of disodium dihydrogen-ethylene-diamine-

tetra acetate and 0.05 g of MgCl2.6H2O in water and dilute to 1 litre. Standardise
against standard Ca-solution.
Procedure
● Weigh 5 g of soil sample in a 250 ml conical flask, and add 100 ml of saturated CaSO4
solution.
● Shake for 5 minutes on a mechanical shaker and filter.
● Pipette out 5 ml of the extract into a 100 ml conical flask and dilute to about 25 ml with
distilled water.
● Add 0.5 ml of the NH4Cl-NH4OH buffer and 3-4 drops of the EBT indicator.
● Titrate with the standard EDTA solution until the colour changes from wine red to
blue.
● Titrate in a similar way 5 ml of the saturated CaSO4 separately.
Calculations
Weight of soil taken =5g
Total volume of extract = 100 ml
Let volume of EDTA used for titration of x ml of gypsum solution be A ml (say)
and volume of EDTA used for titration of y ml of sample aliquot be B ml (say)
Therefore, meq. of Ca/l in gypsum solution
FG A IJ × 0.01 × 1000 = P meq./l
= H XK
Meq. of Ca/l in sample solution
FG B IJ × 0.01 × 1000 = Q meq/l
= H YK
Total meq. of Ca remained in soil after addition of 100 ml gypsum
(P – Q) × 100
= = 0.1 (P – Q)
1000
Now 5 g soil contains 0.1 × (P – Q) meq.
0.1 × (P − Q)
Hence 100 g soil contains × 100 meq.
5
= [2 × (P – Q)] meq./100 g
Thus 1 kg soil requires [20 × 20 × (P – Q)] mg Ca
= 400 × (P – Q) mg Ca.
400 × (P − Q) × 2.24 × 10 6
2.24 million kg soil requires
10 6
= 896 (P – Q) kg Ca.
Now 40 kg Ca is obtained from 172 kg gypsum
172 × 896 × (P − Q)
So, [896 × (P – Q)] kg Ca is obtained from
40 × 1000
= [3.85(P – Q)] tons of gypsum
Thus gypsum requirement of the soil = [3.85 × (P – Q)] tons/ha.
SOIL CHEMISTRY 133

3.23 DETERMINATION OF LIME POTENTIAL

Soil pH measurement is usually affected by a number of factors viz. salt concentration,
soil-water ratios, suspension effect etc. The use of 0.01(M) CaCl2 solution yields stable readings
in pH-measurements. The hydrogen ions in the soil system are distributed between solid and
liquid phases as follows. The dynamic equilibrium may be represented as follows.
H+Exch H+Soln. ...(3.23.1)
+
The H ions in the soil solution constitutes the ‘active acidity’ and are measured directly
as soil pH values. On the other hand the adsorbed H+ ions held on exchange sites are not subject
to pH measurements are termed as ‘reserve acidity’, both the forms contribute to soil acidity.
Thus soil pH does not reflect the total acidity. However a suitable index which takes into ac-
count the reserve acidity of soil is the lime potential, which is calculated as follows:
Lime potential = pH – ½ pCa ...(3.23.2)
where, pCa = – log (Ca ) 2+ ...(3.23.3)
The lime potential is a very reliable estimate to predict the buffering capacity of soils.
Smaller the value of lime potential, greater will be the buffering capacity of the soil.
Schofield and Taylor (1955) suggested the use of ion acitivity ratios for determination of
soil acidity. Consider a cation C with valency ν. In dilute solution in equilibrium with a soil the
acitivity of hydrogen ions divided by the activity of C ions will be constant. The activity function
depends on the valency of ions concerned.
aH+
Therefore = constant ...(3.23.4)
(ac ) 1/ V
If the soil exchange complex is saturated with both H+ and Ca2+ ions at equilibrium
Schoefield’s ratio law states :
aH +
= constant (K) ...(3.23.4)
aCa 2 +
The negative logarithm of (K) is called the lime potential
a LM 1
– log H2++ = – [log aH+ – log aCa2+ ] = – log aH + − log aCa 2 +
OP
i.e.
aCa N 2 Q
...(3.23.6)

FG 1 IJ = LM(− log a 1 OP FG 1 IJ
H
= − log aH + +
2
log aCa 2+
K N H+ ) −
2 Q H
(− log aCa 2 + ) = pH − pCa
2 K
...(3.23.7)
Principle
In practice soil is shaken with a calcium chloride solution of known strength (1 : 2
soil:solution ratio) and activity of Ca2+ ions and the pH of the suspension is measured. Lime
potential is calculated as; (measured pH – 1.14), 1.14 being the value of ½ pCa for 0.01(M) CaCl2
solution. The use of 0.01(M) CaCl2 solution as an extractant simulates the electrolyte level of
non-saline soil at optimum field water content and more so the H+ ion environment existing in
the soil solution-plant root system.
Reagents
● 0.01(M) CaCl2 solution; Dissolve 1.3 g of anhydrous CaCl2 in water and dilute to one
litre.
● Buffer solutions for pH measurement.
134 PHYSICAL AND CHEMICAL METHODS IN SOIL ANALYSIS

Procedure
● Weigh accurately 10 g of soil in a conical flask and add 20 ml CaCl2 extracting solution
into it.
● Shake for half an hour and measure the pH of the suspension using a pH meter.
Calculation
Lime potential = pH – 1.14

3.24 AVAILABLE SULPHUR DETERMINATION IN SOIL

Turbiditmetric Procedure; Monocalcium Phosphate Extratable S (Ensminger, 1954).
Principle
Sulphur (S) occurs in soils usually as sulphites, sulphates, sulphides and in organic
compounds. However, the most accessible form is ‘sulphate’ (SO4=). The turbidimetric procedure
is widely used in the estimation of available S in the soil due to its rapidity. However, erroneous
results are obtained in case the soil is rich in organic matter. Soil is shaken with a solution of
monocalcium phosphate, containing 500 ppm P. The phosphate ions displaces the adsorbed
sulphate. The calcium ions depresses the extraction of soil organic matter, thus eliminating
contamination from extractable organic S. The method extract soluble SO42– plus a fraction of
adsorbed SO42–. The filtrate is then analysed for S by the turbidimetric procedure. In this method
the filtrate is treated with barium chloride in the presence of gumacacia solution, and the
turbidity produced by the precipitation of sulphate as barium sulphate is measured
colorimetrically. Gumacacia helps in preventing rapid settling of barium sulphate precipitate.
Ca(H2PO4)2 . 2H2O → Ca+ + 2H2PO4– + 2H2O ...(3.24.1)

OP
Soil colloid SO 4 + 2H 2 PO 4 − → Soil Colloid
OP H 2 PO 4

+ SO 4 2−
PQ PQ H 2 PO 4
...(3.24.2)

BaCl2 → Ba2+ + 2Cl– ...(3.24.3)

Ba2+ + SO42– → Ba(SO4) ...(3.24.4)
precipitate
Reagents
● Monocalcium phosphate solution; Dissolve 2.18 g of Ca(H2PO4)2 . 2H2O in distilled
water, and dilute to one litre.
● Barium chloride; Grind BaCl2 crystals in a mortar, until they pass through a 30 mesh
sieve and are retained on a 60 mesh sieve. The crystals are added to sulphate solution
in the solid state as crystals of definite size and not as solution.
● Gum-acacia solution, 0.25%; Dissolve 0.25 g gum-acacia in distilled water, and dilute
to 100ml.
● Standard S solution; Dissolve 0.5434 g (AR-grade) potassium sulphate in distilled water
and dilute to one litre. This gives 100 ppm stock solution of S.
Procedure
● Weigh accurately 20 g of air dried soil and transfer it to a 250 ml Erlenmeyer flask.
● Add 100 ml of monocalcium phosphate extracting solution and shake for half an hour.
Filter through Whatman No.42 filter paper under suction.