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 Learners Guide to

Soil Analysis

B.GOKILA
S. SIVAGNANAM
BHARGAVA RAMI REDDY CH

2017
Rs. 145/-

Published by: Thannambikkai Publication, Coimbatore.

All rights reserved. No part of this publication may be reproduced, stored in a retrieval system or
transmitted in any form or by any means, electronic, mechanical, photocopying, recording or
otherwise without prior permission of the publisher or the copyright holder.

This book contains information obtained from authentic and high regarded sources. reasonable
efforts have been made to publish reliable data and information, but the author/s, editor/s and
publisher cannot assume responsibility for the validity of all materials or the consequences of
their use. The author/s, editor/s and publisher have attempted to trace and acknowledge the
copyright holders of all and acknowledgements to publish in this form have not been taken.
Composed, Designed and Printed at Sree Kumaran Computers, Coimbatore.
Preface

This book on soil analysis is framed based on the syllabus for Agricultural Science
and accordingly it will serve as a text book for students of Agricultural,
Horticultural Science, Home Science and Environmental Science besides the basic
sciences. The main frame work of this book consists of 4 chapters and 40 sub topics
and five annexure. The contents of this book are arranged in such a way that it
would help the readers to understand the basic concepts and techniques of the
different analytical methods. The chapters are divided into four comprising of
analytical chemistry, soil physics, soil chemistry and fertilizer & manure
analysis. A brief account of basic theories involved, reactions, materials
required, reagents and their preparations, detailed procedure for estimation
and calculations are explained in detail.

Sincere efforts have been made to compile this book and the analytical
methods are depicted in a self-explanatory manner to readers. The authors hope
that the researchers and students of Agriculture and allied faculties would find
this book useful in learning different techniques and usefulness in their
laboratory analysis. We would be much thankful for receiving comments and
suggestions, if any, on any aspect of this publication, which would definitely
help in improving the future editions of this book.

The authors express their sincere thanks to all scientists and friends for their
contribution to our inspiration and knowledge.

Authors
 CONTENTS

Chapter Title Page

No. No.
1 Analytical Chemistry

1.1 Introduction 1

1.2 Weighing in an analytical balance 9

1.3 Calibration of Glasswares 13

1.4 Use of indicators in different situations 17

1.5 Preparation of primary standard solutions 21

1.6 Preparation of secondary standard solutions 24

1.7 Alkalimetry (Standardisation of a base) 29

1.8 Oxidimetry (Standardization of KMnO4) 32

1.9 Dichrometry (Standardization of Ferrous Sulphate) 35

1.10 lodimetry (Determination of copper) 37

Complexometry (Estimation of calcium and magnesium by Versenate
1.11 39
method)
1.12 Precipitometry (Estimation of chloride) 42

1.13 Gravimetry (Estimation of Sulphate) 44

2 Soil Physics

2.1 Collection and Preparation of Soil Samples for Laboratory Analysis 46

2.2 Determination of Bulk Density, Particle Density and Pore Space 50

Determination of infiltration of soil (Double Ring Infiltrometer
2.3 53
Method)
2.4 Determination of Aggregate Size Distribution of Soil 58

2.5 Estimation of Soil Water Potentials Using Tensiometer 62

2.6 Determination of Field Capacity of Soil 64

2.7 Determination of Saturated Hydraulic Conductivity of Soil 66

2.8 Determination of the Pore Space of the Soil (Buckner Funnel Method) 73
Determination of Moisture Content Under Different Suction Levels
2.9 74
(Pressure Plate Method)
2.10 Measurement of Soil Temperature 76

2.11 Estimation of Moisture Content in Soil 81

Determination of Soil Texture (International Pipette Method/ Robinson's
2.12 82
Pipette Method)
3 Soil Chemical Analysis
3.1 Determination of Soil pH and Soil Electrical Conductivity 88
Estimation of available Nitrogen in soil by Alkaline Permanganate
3.2 93
method (Subbiah and Asija, 1956)
Estimation of Available Phosphorus in Soil by Olsen and Bray No.1
3.3 95
Method (Olsen et al., 1954)
3.4 Estimation of available Potassium in Soil (Stanford and English, 1949) 99

3.5 Estimation of available Sulphur in soil (Williams & Steinbergs, 1959) 101

3.6 Estimation of Available Micronutrients (Lindsay and Norvell, 1978) 104

4 Fertilizer / Manure Analysis

4.1 Fertilizer Sampling Technique 107

4.2 Estimation of Nitrogen in Urea 110

4.3 Estimation of NH4 – N and NO3 – N in Ammonium Nitrate 113

Estimation of Water- Soluble P in Superphosphate/Dicalcium Phosphate
4.4 116
/ Rock Phosphate
Estimation of Citric Acid-Soluble P in Dicalcium Phosphate / Rock
4.5 119
Phosphate
4.6 Estimation of Potassium in Potassium Chloride / Potassium Sulphate 121

4.7 Estimation of Nitrogen in Organic Manure / Oil Cake 123

4.8 Estimation of Phosphorus in Organic Manure / Oil Cake 125

4.9 Estimation of Potassium in Organic Manure / Oil Cake 127

 1. ANALYTICAL CHEMISTRY

1.1 Introduction

Analytical chemistry deals with qualitative and quantitative analyses. The former
shows the nature of elements or ions present in a given substance. The latter determines
the quantity of individual elements or ions or compounds present in a given substance.
The results of quantitative analysis are usually expressed in percentage or often expressed
in percentage of their respective oxides.

A variety of methods, viz., chemical, physical and physico-chemical can be used

for qualitative analysis. In chemical methods of qualitative analysis, the element or ion to
be detected is converted into a new compound which has specific properties on the basis
of which its formation can be elucidated.

A distinction is made between macro, micro, semi micro and ultra micro methods
of qualitative analysis which depends on the amounts of substances used for the
analytical reactions.

In macro analysis relatively large amounts of substance (0.5 -1.0 g) or solution

(20 - 50 ml) are used. The reactions are carried out in ordinary test tubes, beakers or
flasks. Precipitates are separated from solutions by filtration through filter papers.

The amount of substances used in micro analysis is usually about 1/100th of the
amount involved in macro analysis work. Semi -micro analysis occupies an intermediate
position between the macro and micro methods. Semi - micro analysis has a number of
advantages over macro analysis and gives equally reliable results with careful working.
Ultramicro analysis is used for investigation of less than 1 mg of substance. The various
analytical operations are performed under the microscope.

Based on the nature of analysis, quantitative analytical methods are grouped as

chemical analysis (inclusive of volumetric, gravimetric and gas analysis),
physicochemical analysis (inclusive of colorimetric, nephalometric, electrochemical,
chromatographic etc.) and physical methods (inclusive of quantitative spectroscopic
analysis, luminescence analysis etc.

1
 Based on the concentration of the element or constituent in the sample to be analysed,
the methods are also grouped as major (1-10 per cent), minor (less than 1 per cent), trace
(1 -100 ppm) and ultra trace (1 ppm).

The most common quantitative analytical methods include (i) volumetric analysis,
(ii) gravimetric analysis and (iii) instrumental methods of analysis,

Volumetric analysis

Volumetric analysis enables us to ascertain the given substance quantitatively by

a careful measurement of the volume of a solution of known concentration required to
react with the solution to be analysed. The substance is allowed to react with a reagent of
known strength and from the volume of the reagent used for the complete interaction of
the substance, the amount of the substance present is determined.

Volumetric methods of analysis are largely employed in the technical work, since
the speed of working and the accuracy are combined with economy of reagents and the
possibility of estimating one or more substances in the presence of others without
effecting any separation. Unlike gravimetric methods, here the final measurement is
made by volume measurement.

The method involves

1. The preparation of one or more standard solutions of accurately known concentration

2. The use of accurately graduated vessels, by which volumes can be rapidly measured.

3. Some means of recognizing the end point, ie., the completion of reaction.

Advantages

1. Economy of time; tidious process of weighing is not always required. Volumetric

methods are more rapid.

2. Minute quantities of substances which would be very difficult to weigh accurately,

can be worked out easily.

2
Titrimetry

Titrimetry is based on the fact that the product of volume and concentration of
one reacting substance is equal to the product of volume and concentration of the second
reacting substance involved in the titration, which can be shown in the reaction equations:

V 1N1 =V2 N2

Where

V1 - Volume of the first reacting substance

N1- Concentration of the first reacting substance

V2 - Volume of the second reacting substance

N2 - Concentration of the second reacting substance.

Titrimetry involves determining the quantity of reagent which exactly

corresponds to the above reaction equation and is chemically equivalent to the substances
being determined.

1. Acidimetry and Alkalimetry

These are the methods where the interaction between hydrogen (H+) and hydroxyl
(OH') ions occur. If an acid is added to the base (or vice versa), the pH of the solution
will change during the course of titration and so by using a pH sensitive indicator like
methyl orange, methyl red or phenolphthalein, the end point can be determined.

2 NaOH + (COOH)2 2COONa + 2H2O

2 Na2CO3 + 2 HCI 2 NaCI + H2O + CO2

2. Oxidation - reduction or redox titrations

These involve the change in the oxidation state of the reacting substances. The
commonly used, oxidising agents include potassium permanganate and potassium
dichromate, while the reducing agents are ferrous ammonium sulphate and oxalic acid.
Based on the reacting substances involved, redox titrations are grouped into four
categories as follows:

3
 i) Permanganate titration (Permanganametry)

ii) Dichromate titrations (Dichrometry)

iii) Iodine titration (lodimetry)

iv) Copper reduction titrations

3. Precipitation titrations

The reacting constituents precipitate during the course of titration. When that
substance is completely precipitated, the additional drop of the titrant will react with the
indicator making the end point. Since the titrations involve the silver compounds they are
also called as argentimetric or argentometric titrations.

AgNO3 + NaCI AgCl + NaNO3

AgNO3 + KCNS AgCNS + KNO3

4. Complex formation or complexometric titrations

The reacting constituents form undissociable stable complex thereby the

concentration of a particular ion in the solution gradually decreases. When all the ions
have been completely complexed, any additional drop of the titrant will react with the
indicator marking the end point.

Common terminologies used in volumetric analysis -

1. Standard solution

A solution of known concentration or strength is called as standard solution. It

contains a weight of the solute in a known volume of the solution.

2. Formality or formal concentration

The 'formality' (F) of a solution expresses the number of gram-formula weights of

solute present in one litre of solution. This term also gives the number of milliformula
weights of solute in one ml of the solution.

4
3. Molarity or Molar concentration

The 'Molarity' (M) of a solution defines the number of gram-molecular weights

(or moles) of a species in 1 litre of solution or the number of millimolecular weights in
one ml of solution.

The molar concentration and the formal concentration of a given solution will
frequently differ. Thus the solution that results when one gram-formula weight of oxalic
acid (90.04 g H2C2O4) is dissolved in sufficient water to give 1 litre. Because this solute
undergoes dissociation to the extent of approximately 22 per cent, however, the species
concentration prepared by dissolving 1 gram-formula weight of sodium chloride (58.44 g) in
sufficient water to give 1 litre of solution will be 1 F with respect to NaCI, by definition;
the concentration of both Na+ and Cl- will be 1 M. The molar concentration of the species
NaCI will be zero. This is the general situation for solutions of strong electrolytes.

From these examples, it is clear that quantitative information is needed with regard to
the composition of the solute in solution before its molar concentration can be specified. On
the other hand, formal concentration may be calculated from the specifications for the
preparation of solutions and the formula weight of the solute.

4. Normality or Normal concentration

The 'normality' (N) of a solution expresses the number of milliequivalents of

dissolved solute contained in 1 milli litre (Thus, a 0. 2 N solution of silver nitrate contains
0.20 milli equivalent of this substance in each milli litre of the solution).

5. Percentage concentration

Chemists frequently denote the concentration of solutions in terms of percentages

and this may be done in several ways: Three of the common methods are as follows:

Weight percent = Weight of Solute

--------------------- × 100
Weight of Solution
Volume percent = Volume of Solute
----------------------- × 100
Volume of Solution

5
 Weight - Volume percent = Weight of Solute
--------------------- × 100
Volume of Solution
Weight - Volume per cent is frequently used to indicate concentration of dilute
aqueous solutions of solid reagents; thus a five per cent silver nitrate solution is prepared
by dissolving five gram of silver nitrate in water and diluting to 100 ml.

6. Parts per million

In expressing the concentrations of very dilute solutions, percentage compositions

become inconvenient to use because of the number of zeros needed to place the decimal
point. Under these circumstances, the concentration is more conveniently expressed in
parts per million (ppm). This term is defined by the equation

Ppm = Weight of Solute

--------------------- × 10,00,000
Volume of Solution

Thus an aqueous solution containing 0.003 per cent nickel by weight contains
3 ppm nickel.

7. Titer

The titer of a solution is the weight of a substance that is chemically equivalent to

one ml of that solution. Thus, a silver nitrate solution having a titer of 1 mg of chloride
would contain just enough silver nitrate in each milli litre to react completely with that
weight of chloride ion. The concentration of a reagent to be used for the routine analysis
of many samples is advantageously expressed in terms of its titer.

8. Milliequivalent

A milliequivaient represents 1/1000 of an equivalent weight. The equivalent

weight of an element or radical is the weight in grams of the element or radical required
to displace or to unite with one atomic weight of H+ or to combine with one atomic
weight of Cl- or of any other univalent element. The equivalent weight of H+ is 1.008 and
that of Cl- is 35.46, since their respective atomic weights are 1.008 and 35.46.

6
The equivalent weight of K is 39.1 because it takes one atomic weight of K to combine
with one atomic weight of Cl to form KCI. The equivalent weight of the ammonium
radical (NH4) to combine with 35.46 gm Cl which is one atomic weight of Cl. The
equivalent weight of Ca is 20 because it takes two atomic weights of Cl for one atomic
weight of Ca which is 40 to make up the compound CaCl2.

Example

100 grams of soil, when treated with NH4CH3COOH gave up the following cations;
8.064 mg of H; 60 mg of Ca; 8 mg of Mg and 78.2 mg of K. In terms of milliequivalents
per 100 grams of soil, each cation may be calculated as follows:

8.064 / 1.008 = 8 milliequivalents for H; 60 / (40 / 2) = 3 m.e. for Ca; 8 / (24 / 2) = 0.67
m.e. for Mg; 78.2 / 39,1 - 2 m.e. for K. The total cation exchange capacity for the
particular soil will thus be 8+3+0.67×2=13.67 milliequivalents/100 gram of soil.

9. Equivalence point

When one substance is allowed to react with another substance, the stage at which the
reaction between the two substances exactly comes to an end is termed as equivalence point,
ie., in titration the reagent is used not in excess but in a quantity which exactly corresponds to
the reaction equation and is chemically equivalent to the substance being determined.

10. End Point

This represents the point of completion of reaction, when a solution is titrated

with another solution. The end point is generally located at, or as close as possible to the
equivalence point of the titration.

Cleaning Solution

Except in special instances, all ordinary glasswares and volumetric equipments

are best cleaned with 1-2 per cent detergent solution (teepol or soap solution or
sometimes vim can also be used).

Mechanical action, such as shaking or scrubbing with a bottle brush, increases

cleaning efficiency. Sometimes, 1-2 per cent warm trisodium phosphate (Na3PO4)
solution is also used.

7
 When detergents and trisodium phosphate are ineffective, cleaning solution
(H2SO4 - K2 Cr2 O7) may be used. It is very effective in cleaning glasswares containing
deposits of organic materials. It is prepared by adding about 10 g of potassium
dichromate to about 200 ml of hot (100°C) concentrated sulphuric acid in a large beaker
and stir it for few minutes to allow K2Cr2O7 to go into solution. The mixture is then
allowed to cool to about 50°C and is decanted for storage into 500 ml wide-mouthed
glass-stoppered bottle, leaving undissolved crystals behind.

To use cleaning solution, the apparatus should first be rinsed with water.
The cleaning solution is poured into the apparatus so as to wet all parts. The apparatus is
left to stand for a few minutes to give the time for cleaning the solution to act, after which
the excess cleaning solution is poured back into storage bottle, and the apparatus is
cleaned J- with water thoroughly and rinsed with distilled water.

Reading the position of meniscus

If the liquid surface is concave for clear liquids, the bottom of the meniscus can
be precisely located avoiding parallax errors. In case of difficulty, a black / white reading
card may also be used.

In the case of dark or deeply coloured solution (as KMnO4 or iodine) the bottom
of the meniscus may not always be visible and locatable by the procedure above. In that
case reading to the top of the meniscus can be followed.

8
 1.2 Weighing in an analytical balance

Almost every chemical analysis requires the accurate weighing of samples. If the
samples are not weighed accurately, wrong results will be obtained. Proper use of
analytical balance is therefore the first step to be learnt for all the elementary analysis to
be carried out in analytical chemistry.

Physical balance

Physical balance is used to measure the mass of an object In the SI system, mass
of an object is measured in kilograms and weight is measured in Newton.

Materials required

1. Physical balance

2. Weight box with fractional weights

Description

The physical balance consists of an aluminium (a light metal) beam supported at

its centre on a knife edge made of agate (or) hardened steel. This knife edge points
downwards and rests on a small flat platform also made of agate hardened steel.

The platform is fixed to the top of a similar pillar which can be moved vertically
up and down by turning a knob located at the front of the balance. There are two smaller
knife edges pointing upward - one on either end of the beam at equal distance from the
centre of the beam. Two strips carrying identical scale pans are suspended - one from
each of the smaller knife edge. A pointer is fixed to the centre of the beam and is
perpendicular to the beam.

The lower end of the pointer moves on an ivory scale fixed at the bottom of the
pillar. During measurement of mass of an object as the beam swings up and down on
each side, the pointer swings from side to side on the ivory scale. There are two adjusting
screws on either ends of the beam. These screws are adjusted to make the pointer swing
equally ' on either side from the centre of the ivory scale.

The physical balance is placed in a wooden box with glass windows to protect it from
dust and air currents which can cause disturbances during measurement. There are two
levelling screws at the bottom of the balance to ensure that the pillar is exactly vertical.

9
 A plumb-line is provided near the centre of the beam in order to check whether
the pillar is vertical, before the balance is used for mesurement. When not in use, the
pillar is lowered so that it rests on two metal supports one on either side. This minimises
the wear and tear of the knife edges which can lead to a reduction in the sensitivity of the
balance and lowering of accuracy.

Procedure

x The two pans are gently cleaned off dust with a soft brush.

x The pillar is made vertical and the beam horizontal by adjusting the levelling
screws at the base of the balance. The levelling is verified by the plumb line.

x The pillar is raised so that the pointer swings on the ivory scale. The adjusting
screws at either end of the beam are adjusted to ensure that the pointer swings
equally on both sides of the central division on the ivory scale.

x The zero resting point of the balance is determined.

x The object, whose mass is to be determined, is placed in the left pan of the
balance.

x Weights are added to the right pan in descending orders of magnitude until the
pointer oscillates equally on both sides of the ivory scale. The resting point is
determined.

x If this resting point is equal to the zero resting point, the total weights in the right
pan give the weight of the object.

x If the zero resting point is greater than the resting point, 10 mg is added to the
right pan. If it is less than the resting point, 10 mg is removed from the right pan
and the new resting point is determined.

x The weight corresponding to that resting point which is nearer to the zero resting
point gives the weight of the object correct to a centigram.

x The physical balance is a very sensitive instrument and should be handled with
care in order to maintain its accuracy. The following points should be kept in
mind while using this instrument.
10
 x The weights must be handled with forceps only. While adding or removing
weights the pillar should be lowered and the beam should be always at resting
position.

x The standard weight must be added in descending order.

x The doors of the balance must be closed while observing the turning points to
avoid disturbance due to air currents.

II. Chemical balance

A chemical balance is used to measure the mass of an object more accurately

without much oscillation.

Materials required

1. Chemical balance

2. Weight box with fractional weights

3. Rider

General rules for weighing

x Check whether the balance is in order by releasing the control knob gently and
observe that the pointer swings evenly on both the sides.

x Before the commencement of weighing, the balance must be adjusted to swing

evenly with the rider placed directly over the centre of the beam.

x The objects to be weighed must be at room temperature so as to avoid air

convention currents.

x The object to be weighed must be placed on the left hand scale pan and the
weights on the right hand scale pan.

x The object to be weighed should be placed on a watch glass, scoop (or) weighing
bottle and should not be directly kept on the balance path.

x The balance must be in arrest position when adding any weight to the balance pan
(or) removing anything from the pan. The balance must be assessed when the

11
 rider is moved along the beam. The weight must be handled using the forceps
supplied for that purpose. The same forceps is used for handling the rider on the
hook (or) taking it off.

x The side doors of the balance case must be closed while weighing.

x All weighing must be done on the same balance and with the same set of weights.

x After weighing is over, the weight must be placed in their proper place in the
weight box. The rider must be removed and kept inside the weight box and the
balance case must be closed.

The rider

The rider is a small piece of wire whose weight is accurately known (10 mg). It is
bent to a shape so that it will sit evenly across the beam of the balance. It may be moved
by means of a hook to various positions on the beam in order to record weights lower
than 0.01 g.

The balance is adjusted to swing evenly with the rider directly over the centre of
the beam. In this position it has no effect on the swing of the balance. If moved to the last
point on the beam directly over the right hand scale pan, it exerts its full weight of 10 mg
over that pan.

The intermediate positions indicated by the graduations from 1 to 10, representing

successive increments of 1 mg (0.001g). i.e., third decimal place in gram. The fourth
decimal place is obtained by further subdivisions of the milligram graduations. If each
mg is further subdivided into 5 parts, then each sub division represents one - fifth of
0.001 g = 0.0002 g (or) 0.2 mg i.e., the precision level of the balance.

Weighing procedure

The object to be weighed must be placed in the left pan and the weights in
descending order on the right pan upto 0.01 g short of the actual weight of the object. The
third and fourth decimal fractions of a gram are obtained by moving the rider between 0
and 10 on the beam till the pointer swings equally on either side of the middle of the
scale. The actual weight of an object is measured by adding this fraction of weight.

12
 1.3 Calibration of Glasswares

Calibration of pipette

Principle

Volume of water differs according to the temperature. By determining the actual

volume of water delivered at a particular temperature by referring to the table, the
calibration correction is made for pipette and burette. Since alkali corrodes glasswares,
the burette and pipette should be of alkali resistant glass.

Volume of one gram of water at different temperatures given in the following table

Volume
Temp. °C Temp.° C Volume (ml) Temp.°C Volume (ml)
(ml)

18 1.0025 24 1.0036 30 1.0051

19 1.0026 25 1.0038 31 1.0054

20 1 .0028 26 1.0041 32 1.0056

21 1.0030 27 1.0043 33 1.0059

22 1 .0032 28 1 .0046 34 1.0062

23 1.0034 29 1.0048 35 1.0066

Apparatus required

1. 10 mL pipette

2. 125 ml or 100 ml conical flask

3. Beaker

4. Thermometer

5. Burette and Balance

13
Procedure

x Clean the pipette, which is to be calibrated

x Pipette out distilled water, the temperature of which is already noted

x Deliver the contents of the pipette into an already weighed conical flask and
determine the weight again

x From the weight of water delivered, find the actual volume of water from the
table. The difference between the nominal volume and actual volume gives the
calibration correction which should be added to the nominal volume.

x Repeat the process and calculate the average which should be used as the
calibration correction for the pipette in all future works.

Calculation

Suppose a 10 mL pipette is calibrated at 28° C

Empty weight of the flask = yg

Weight of flask + water delivered by the pipette = xg

Therefore, weight of water delivered by the = (x-y) g

pipette

Now, pipette volume = (X-y) (1.0046 mL)

Calibration correction = (x-y) (l.0046) / 10.0 mL

True volume = Nominal volume + calibration

correction

B. Calibration of burette

Procedure

x Take a 50 mL burette and clean well

x Check for any leakage and fill it with distilled water and remove air bubbles, if any

x Bring the water level to zero mark

x Leave it for 30 seconds and drain the water drops if any at the tip just by touching

14
 x Take a clean dry 100 mL conical flask and determine its weight

x Place it under the burette and drain exactly 10 mL of water (0-10 mL) and
determine its weight.

x Calculate the weight of water (i.e. 10 mL)

x Again drain 10 ml of water into the conical flask (10-20 mL) and determine the
weight as before Repeat this process for the entire 50 mL. From this calculate the
actual volume of water by referring to the table (volume of water for various
temperatures).

x Repeat the whole procedure for two or three times and find the average of the
determinations.

x Plot the average calibration corrections (in hundredth of a milli litre) for each of
the given intervals. Draw a curve through the points. Depending on the initial and
final titre values, suitably select the calibration correction from the table prepared
and adopt for all the future works. eg. Water temperature 30°C.

Weight of the
Weight
flask (g) Actual
of the
Burette Approx. volume Correction Total
No. water
reading (mL) With delivered Factor Correction
delivered
Empty delivered (ml)
(g)
water

(f) = (g) =
(a) (b) (c) (d) (e) (h)
(e-d) (f×1.005)

1 0-10 10.0 60.00 70.00 10.00 10.05 +0.05 0.05

2 10-20 10.0 60.00 70.00 10.00 10.05 +0.05 0.10

3 20-30 10.0 60.00 70.00 10.00 10.05 +0.05 0.15

4 30-40 10.0 60.00 70.00 10.00 10.05 -0.05 0,20

5 40-50 10.0 60.00 70.00 10.00 10.05 +0.05 0.25

+0.25 mL

15
 Empty weight of flask = 50 g

The total calibration correction for burette = + 0.25 mL

Average calibration correction = + 0.25 / 50.0

= +0.005 per mL

= + 0.0005 per 0.1 mL

If the actual volume thus calculated is higher than approximate volume (marked
on the burette) then the correction factor is given with a '+' symbol. If it is lesser, then '-'
symbol is given.

C. Calibration of volumetric flask

Procedure

x Clean the volumetric flask (25 mL), dry it in the oven at 105°C and cool it in a
desiccator

x Weigh the dry empty flask

x Fill the flask to the mark with water whose temperature is known

x (Outer surface of the flask should not be moistened while filling)

x Find the weight of water contained and from this weight find the true volume and
calibration correction using temperature of water from the table.

x Repeat this process and find the average calibration correction.

16
 1.4. Use of indicators in different situations

Indicators

Indicators are chemicals used in volumetric analysis to induce the end point
(equivalence point) of a chemical reaction.

Self indicators

One among the two solutions involved in the titration will impart a colour change
at the end point. In the titration of ferrous sulphate with potassium permanganate, no
special indicator is required because after the permanganate has oxidized all the ferrous
sulphate and are transformed to the ferric sulphate, further addition of an excess drop of
the reagent will impart a permanent faint pink colour to the solution which indicates the
end point.

Internal indicators

These are directly added to the solutions being titrated and are acted upon only
after the reaction is over. Examples of internal indicators are phenolphthalein, methyl red,
methyl orange, starch, potassium chromate, diphenylamine, etc.

External indicators

These are not directly added to the solution since they are immediately acted upon
by the reagents and so a few drops of the solution are taken out of the titration flask from
time to time and tested outside with the indicator solution. For completion of the reaction,
eg. Potassium ferricyanide in the estimation of iron by titration with potassium dichromate.

Mixed indicators

These are used in the titrations where a single indicator fails to show the end point
correctly. In titrations involving weak acid vs weak base, anions of weak acids such as
bromic acid, mixtures of acid/mixtures of bases, polyprotic acids such as phosphoric acid,
etc. the end point cannot be easily detected by using a single indicator. In such occasions,
one indicator is mixed with another indicator or one indicator is mixed with an organic
dye, either one or both of them may be sensitive to pH. Such mixtures of indicators
enable us to detect the end point more accurately even with a narrow change in pH.

17
 eg. Double indicator: Bromocresol green + Methyl Red

Red in acid medium and green in alkaline medium

Methyl green and Phenolphthalein = pH 8.4 - 8.8

Red-violet at pH < 8.4, grey pale blue at pH 8.4 - 8.8 and pink above pH 8.8.

Universal indicators

Mixture of two or more indicators. These are employed to determine the

approximate pH range of a solution. They are available in two ranges viz., narrow where
the colour change is in the pH range of 0.2 to 0.4 units and wide where the colour
changes in two pH units say 6-8, 8-10. The former produces colour change, when pH
varies widely and the latter produces colour change into narrow range. These are not
employed in quantitative analysis.

pH / Test papers

These are filter papers impregnated with indicators. They serve a similar purpose
as that of universal indicators. They are also in two ranges namely narrow and wide.

Indicators may also be neutralisation indicators (Acid - base) eg. Methyl Red;
Redox indicators eg, Diphenylamine; Metal ion indicators eg. Murexide, Eriochrome
black-T; Adsorption indicators eg. Potassium chromate.

Theory of indicators

Ostwalds (1984) theory

Also referred to as ionic theory of indicators. According to this theory,

neutralisation indicators are "weak organic acids or bases in which undissociated
molecules differ in colour from their ions".

eg. Phenolphthalein is a weak acid which does not dissociate in acid medium and
hence no colour with acid solution. In alkali medium the molecules dissociate and
combine with alkali to produce a pink coloured salt. Similarly methyl orange which is
yellow in unionised condition produces red colour when ionised.

18
Chromophore theory

According to this theory, the colour change of indicators is not due to ionisation
but due to change in the structure of indicators. The colour change of indicators is due to
some intra molecular rearrangement, which changes the structure of the indicators.

The colour of organic compounds is due to the presence of chromophores in their

molecules. These chromophores may be radicals (atomic groups) or group of double
bonds. These include

i. Nitro groups : O = N = O which can be converted into O = N-NH- group

ii. Azo group : - N + N - which may be converted into N - NH – group

iii. Quinoid group transformed from benzenoid

iv. Carbony! group C =O

v. Double bonds close to each other

The colour of organic compounds is also due to the presence of auxochromes. These
cannot themselves produce colours but when present along with chromophores they
augment the action of the latter and deepen the colour produced by them.

Examples of auxochromes -OH2- NH2, OCH3-N (CH3)2 etc.

According to this chromophore theory the colour change in indicators is the result
of tautomerism which is a reversible inter conversion of isomeric forms. Due to this
intramolecular regrouping, when the structure changes with the formation or
disappearance of groups which influence colour namely chromophores and auxochromes,
the colour of indicator also changes. The isomers, involved in tautomerism are called as
tautomers,

eg. Paranitrophenol.

The formation of quinoid nucleus under alkaline condition is the cause for the
change of colour of paranitrophenol under alkaline condition. When it is acidified the
equilibrium is shifted in the opposite direction and the colour changes from yellow to
colourless.

19
Choice of indicators

Two quantitative methods are available for choice of indicators

1. Calculation of indicator error

2. Plotting of titration curves

Calculation of indicator error

The colour of indicator changes over a narrow range of pH and this range depends
only on the properties of the given indicator and does not depend upon the reacting acid
or base. Due to this, the colour change of an indicator generally takes place not at the
equivalence point but with deviation. Such deviation is called as indicator error. It is the
error caused by the difference between the titration exponent of a given indicator and the
pH at equivalence point. Several methods are available for calculating the indicator error.

20
 1.5 Preparation of primary standard solutions

Principle

Standard solutions are solutions of accurately known concentration. There are two
types of standard solutions viz., primary and secondary.

A known quantity of the substance is dissolved in a volumetric flask and the

volume is made up to the mark. Standard solutions prepared in this way are known as
primary standards. This method can be used for the preparation of standard solutions of
substances, which satisfy the following conditions.

i) The substances must be chemically pure, i.e. it must not contain impurities in
amounts which could affect analytical precision (not more than 0.05 to 0.1%). In
other words, the substances must be of known and high degree of purity.

ii) The substance must be stable on keeping, both in the solid state and in the
solution, as other wise its composition would cease to its formula. It must be non
hygroscopic and not easily affected by acid fumes, CO2 etc inside the laboratory.

iii) The gram equivalent of the substance should be as large as possible, so that the
precision in determining the normality of the solution is high.

iv) The substance must withstand fairly high temperature, so that it can be dried at
110°C.

v) The substance must be freely soluble in water.

vi) Preferably solid and crystalline compounds.

Chemicals that can be chosen as primary standards

9 Succinic acid

9 Oxalic acid

9 Potassium hydrogen pthalate

9 Benzoic acid

9 Sulfamicacid

9 Sodium carbonate
21
 9 Borax

9 Sodium oxalate

9 Potassium dichromate

9 Arsenic trioxide

9 Sodium chloride

Reagents required

Na2CO3 AR grade

Apparatus required

1. 250ml volumetric flask with stopper

2. Funnel

3. Chemical balance with weight box or Electronic balance

Example: Prepare 250 ml of 0.1 N Na2CO3

Molecular weight of Na2CO3 = 106g

Valency = 2

Equivalent weight of Na2CO3 = Molecular weight / Valency

106/2 =53

Therefore one equivalent of Na2CO3 = 53 g

53 g dissolved in 1 litre gives 1 N solution

5.3 g dissolved in 1 litre gives 0.1 N solution and

1.325g dissolved in 250 ml gives 0.1 N solution

22
Procedure

9 Weigh 1.325 g of Na2CO3 (AR grade, oven dried at 110°C)

9 Transfer this to a 250 ml volumetric flask using a funnel,

9 Wash the funnel, collecting the washings in the flask

9 Remove the funnel and add about 100 ml of distilled water to the flask

9 Shake the flask well so as to dissolve Na2CO3

9 Add, if necessary, one more installment of 100 mL of water and again shake and
thoroughly mix the contents ensuring complete dissolution of Na2CO3.

9 Make up the volume to 250 ml mark and again shake so as to make uniform
solution

9 Label the solution and preserve it for future work. The label must contain name
of the chemical, strength, date of preparation and the name of person who
prepared the same.

23
 1.6 Preparation of secondary standard solutions

Acidimetry (Standardization of an acid)

Principle

Equal volumes of all acids / bases containing their gram equivalent weights per
litre of the solution react completely with each other.

Na2CO3 + 2 HCI = 2 NaCI + H2O + CO2

Na2CO3 + 2 HNO3 = 2 NaNO3 + H2O + CO2

Na2CO3 + H2SO4 = Na2 SO4 + H2O + CO2

In other words, the product of volume (V1) and normality (N1) of one solution (acid) is
equal to the product of volume (V2) and normality (N2) of another solution (alkali / base)

V1N1= V2N2 ( Law of chemical equivalent)

Example: Prepare 250 mL of 0.1 N HCl

Molecular weight of HCl = 36.45 g

Equivalent weight of Na2CO3 = Molecular weight / Valency

36.45/1 = 36.45 g

Therefore one equivalent of HCl = 36.45 g

Specific gravity of concentrated HCl = 1.18

Per cent purity of concentrated HCl = 35 %

One litre of 0.1N solution of HCl contains 3.645 g HCl

First calculate the volume of cone. HCI that contain this amount of HCI. By
proportion for 35.00 g of HCI, 100 g of HCI solution is required

24
 For 3.645 g HCl = 100 × 3.645 = 10.4 g HCl solution
---------------
35

To convert this weight of acid to volume, it must be divided by the specific

gravity of the acid (1.18), we then have

For 1000 ml 0.1 N HCI, 8.8 mL of cone. HCI is required.

Then for 250 mL (8.8/1000) x 250 = 2.2 ml

(Similarly calculate for HNO3 and H2SO4 the details of specific gravity and per cent
purity can be referred from the respective containers).

Apparatus required

1. 400 mL beaker

2. 10 mL measuring cylinder

3. 50 mL burette

4. 10 mL pipette

5. 250 mL conical flasks and

6. 250 mL measuring cylinder

Reagents required

1. Concentrated HCI

2. 0.1NNa2CO3

3. Methyl orange indicator

Procedure

9 Measure out 250 mL of distilled water into a 400 mL beaker using a 250 mL
measuring cylinder

9 Add 2.2 mL (roughly 2.5 mL) of cone. HCI through a 10 mL measuring cylinder
to the beaker containing water and thoroughly mix it using a glass rod. This is
approximately a 0.1 N solution of HCI.

25
 9 Now, fill up a burette with 0.1 N Na2CO3 (primary standard) and mount it in the
burette stand

9 Pipette 10 mL of the approximately 0.1 N solution of HCI into a 250 mL conical

flask and add a few drops of methyl orange Indicator. Note down the initial
reading of the burette.

9 Titrate the contents of the conical flask against 0.1 N Na2CO3. End point is the
change of colour from pinkish red to straw yellow

9 Tabulate the burette readings and find out the volume of 0.1 N Na2CO3 used in
the titration

9 Repeat the titration until you get concordant titre values.

Now, using the relationship, V1N1 = V2N2 (Law of volumetric analysis), find out
the normality of HCI. If the Na2CO3 consumed is exactly 10 ml, then according to the
above relationship the normality of the prepared HCI solution is 0.1 N. If not, the
normality of the prepared HCI solution may be higher or lower than 0.1 N.

Calculation

Volume of 0.1 N Na2CO3 consumed in the titration = V1 = X mL

Volume of 0.1 N HCI taken = V2 = 10 mL

Normality of Na2CO3 = N1 = 0.1 N

Normality of HCI = N2 = Y (N)

V1N1 = V2N2

X × 0.1 10 ×Y

N2 (Y) = X × 0.1
----------
10

Calculate the volume of water to be added in case the normality is more than 0.1
N or the volume of concentrated HCI to be added in case the normality is less than 0.1 N.

26
If the prepared solution is > 0.1 N

Volume of 0.1 N HCI taken (V2) = 10 mL

Volume of 0.1 N Na2 CO3 used (V1) = X mL

Amount of water to be added for 10 mL = (X-10) mL

Therefore, for (250 –A) mL = (X-10) × (250 –A)

---------------------- mL
10

Where A = volume of the prepared HCI solution already used for rinsing the pipette and
or for titration.

If the prepared solution is < 0.1 N

Volume of 0.1 N HCI taken (V2) = 10 mL

Volume of 0.1 N Na2 CO3 used (V1) = X mL

Normality of Na2 CO3 (N1) = 0.1 N

V1N1=V2N2 Therefore = N2 =Y= V1NI

V2

N1 – N2 = 0.1 – Y= Y (N)

For the preparation of 250 ml of 0.1 N HCl we have taken 2.5 ml

Therefore for Y(N) = 2.5 × Y (250 –A)

---------- × ------------- = mL
0.1 250

Add either the volume of water or the volume of acid as the case may be to the
solution in 400 mL beaker and mix it thoroughly by stirring. Repeat the titration and
calculation processes until the normality of the HCI prepared reaches 0.1 N.

27
Format of titration table

0.1 N Na2CO3 Vs approximately 0.1 N HCI

Burette readings (mL) Volume of titrant

SI. Aliquot Indicator End point
used (mL)
No. taken(mL) Initial (iii) Final (iv) (vi) (vii)
(iv-iii) = (v)

28
 1.7 Alkalimetry (Standardisation of a base)

Principle

Equal volumes of all acids / bases containing their gram equivalent weights per
litre of the solution react completely with each other.

2 KOH + 2 HCI = 2 KCI + 2 H2O

In other words, the product of volume (Vt) and normality (N,) of one solution (acid) is
equal to the product of volume (V2) and normality (N2) of another solution (alkali / base).

V1N1 = V2N2

Molecular weight of KOH = 56 g

Equivalent weight of KOH = Molecular weight (56)

--------------------------- = 56
Valency (1)

Therefore, one equivalent weight of KOH = 56 g

56 g dissolved in 1 litre gives 1 N solution

5.6 g dissolved in 1 litre gives 0.1 N solution

1.4 g dissolved in 250 mL gives 0.1 N solution

Apparatus required

1. 400 mL beaker

2. 10 mL pipette

3. 250 mL conical flasks and

4. 50 mL burette

Reagents required

1. 0.1N KOH

2. 0.1N HCI

3. Phenolphthalein indicator

29
Procedure

9 Weigh 1.4 g of KOH

9 Transfer this to a 400 mL beaker and add 100 mL of distilled water and dissolve it
by stirring with a glass rod

9 Add the remaining 150 mL of water to the beaker. This is approximately 0.1 N
solution of KOH

9 Now, fill up a burette with 0.1 N HCI (secondary standard) and mount it in the
burette stand

9 Pipette 10 mL of approximately 0.1 N solution of KOH into a 250 mL conical

flask and add a few drops of phenolphthalein indicator.

9 Titrate against 0.1 N HCI up to an end point of disappearance of pink colour

9 Note down the burette readings and find out the volume of 0.1 N HCI used in the
titration

9 Repeat the titration until you get concordant titre values

9 Now, using the relationship, V1 N1, = V2 N2, find out the normality of the KOH
solution Calculate the volume of water to be added in case the normality is more
than 0.1 N or the amount of KOH to be added in case the normality is less than
0.1 N

9 Add either the volume of water or the amount of KOH, as the case may be and
mix it thoroughly by stirring

9 Repeat the titration and calculation processes until the normality of the KOH
prepared reaches 0.1 N.

30
Format of titration table

0.1 N HCI Vs approximately 0.1 N KOH

Burette readings (mL)

Aliquot Volume of titrant
SI. Indicator End point
taken used (mL)
No. Initial Final (vi) (vii)
(mL) (iv-iii) = (v)
(iii) (iv)

31
 1.8 Oxidimetry (Standardization of KMnO4)

Principle

Potassium permanganate being an oxidizing agent, can be standardized against

some reducing agents like oxalic acid, ferrous sulphate, ferrous ammonium sulphate, etc.
It being a coloured solution acts as a self indicator. One drop excess of it during titration
will produce a pink colour. As KMnO4 will react only in the presence of acid, equal
volume of dilute H2S04 is added to it. In addition, the reaction takes place at an elevated
temperature (60°C). Always KMnO4 is taken in the burette.

Reactions

2 KMnO4 + 3 H2SO4 K2SO4 + 2 MnSO 4+ 3 H2O + 5 (O)

5 H2C2O4 + 5 (O) 5 H2O + 10 CO2

The equivalent weight of an oxidising agent is the weight of it that supplies one
equivalent of oxygen or 8 parts by weight of oxygen for oxidation. From the above equation
it is clear that 2 molecules of KMnO4 supplies 5 atoms of oxygen or 80 g oxygen.

2 KMnO4 = 5 (O) or 5 x 16 = 80 g oxygen

KMnO4 = 40 g oxygen = 5 x 8 g oxygen = 5 equivalents of oxygen

Molecular weight of KMnO4 = 158

Equivalent weight of KMnO4 = Mol. weight / 5 = 158 / 5 = 31 .6

Therefore, one equivalent weight of KMnO4 = 31 .6 g

31 .6 g dissolved in 1 litre of water gives 1 N solution

3.16 g dissolved in 1 litre of water gives 0.1 N solution

Approximately 1 g dissolved in 300 mL of water gives 0.1N solution.

32
Apparatus required

1. Burette

2. Pipette

3. 250 mL & 10 mL measuring cylinders

4. 400 mL beaker

5. 250 mL conical flask,

6. Burner and

7. Balance

Reagents required

1. AR grade KMnO4

2. 0.1N oxalic acid

3. Dilute H2SO4 (1: 4)

Procedure

9 Transfer exactly one gram of KMn04 to a 400 ml beaker and add 150 ml of
distilled water and dissolve it by stirring with a glass rod.

9 Add another 150 ml of water to the beaker and stir well. This gives approximately
0.1 N solution of KMnO4

9 Fill up a burette with the prepared KMnO4 solution and mount it in the burette
stand

9 Pipette out 10 mL of 0.1 N oxalic acid into a 250 ml conical flask and add to it
10 ml of dilute H2SO4 using a 10 ml measuring cylinder Warm the contents of the
conical flask to about 60°C. Maintain the temperature throughout the titration.

9 Titrate this against approximately 0.1 N KMnO4 solutions taken in the burette till
the appearance of a permanent pale pink colour. Add KMn04 slowly at the
beginning and wait each time until the solution becomes colourless. Continue the
titration to endpoint, with continuous shaking.

33
9 Note down the burette reading and find out the volume of approximately 0.1 N
KMnO4 used in the titration at the end point Repeat the titration till concordant
values are obtained

9 Using the relationship V1N1 = V2N2, find out the normality of KMnO4 solution
prepared and adjust the normality of KMnO4 to exactly 0.1N by suitable dilution
or by addition of salt.

34
 1.9 Dichrometry (Standardization of Ferrous Sulphate)

Principle

Potassium ferricyanide serves as an external indicator producing no blue colour

for a drop of ferrous sulphate at a point of standardisation with 0.1 N K2Cr2O7.

6 FeSO4 + K2Cr2O7 + 7 H2SO4 3 Fe2(SO4) + Cr2(SO4)3 + K2SO4 + 7 H2O

Apparatus required

9 Pipette

9 Burette

9 10 ml measuring cylinder

9 250 mL conical flasks and

9 Porcelain tile

Reagents required

9 AR grade K2Cr2O7

9 Ferrous ammonium sulphate or ferrous sulphate

9 Dilute H2SO4

9 Potassium ferricyanide indicator

Procedure

9 Pipette out 10 mL of the ferrous sulphate solution into a 250 mL conical flask and
add an equal volume of dilute H2SO4.

9 Take a white porcelain tile and put a number of drops of the indicator 2 cm apart
with the help of a glass rod.

9 Fill a burette with 0.1 N K2Cr2O7 and titrate against the ferrous sulphate taken in
the conical flask.

9 From time to time remove a drop of the solution from the conical flask with a
glass rod and touch the indicator drop.

35
 9 If blue colour is formed, continue the titration and continue the test at intervals till
no blue colour is produced.

9 End point is the absence of blue colour formation.

9 This gives a rough estimate preferably the testing may be done at intervals of 1 mL.

9 Now repeat the titration with another 10 m L of the ferrous sulphate solution in a
conical flask to which an equal volume of dilute H2SO4 has been added.

9 Add K2Cr2O7 from the burette 0.5 or 1.0 ml less than the rough reading and test a
drop of the solution from the conical flask.

9 Add further K2Cr2O7 drop by drop and test the solution after the addition of each
drop or two till no blue colour develops.

9 Note the reading and find out the volume of K2Cr2O7 used at the end point.

9 Repeat the titrations until concordant values are obtained.

9 Using the relationship V1N1 = V2N2 find out the normality of the ferrous sulphate
solution prepared.

Calculation

Volume of FeSO4 taken (V1) = 10 mL

Normality of FeSO4 (N1) = X

Volume of 0.1 N K2Cr2O7 = V2

Normality of K2Cr2O7 = 0.1 N

Since V1N1 = V2N2

N1 or X = 0.1
V2 × ----
10

36
 1.10 lodimetry (Determination of copper)

Principle

Cupric salt reacts with potassium iodide and liberates iodine. The iodine liberated
is titrated against 0.1 N Na2S2O3.

2 CuSO4 + 4 Kl 2 K2SO4 +2CuI2

2CuI2 Cu2I2 + I2

I2+ 2Na2S2O3 2Na2S4O6 +2NaI

Apparatus required

9 Burette

9 Pipette and

9 250 ml conical flask

Reagents required

9 Potassium iodide

9 N sodium thiosulphate

9 Freshly prepared starch solution

9 Sodium carbonate

Procedure

9 Pipette out 10 ml of the given solution to a 250 ml conical flask.

9 Add one gram Na3CO3 to neutralise the mineral acids if any Present.

9 Dissolve the precipitate of Cu carbonate by adding acetic acid drop by drop.

9 Add two grams Kl to the flask and titrate the iodine liberated immediately against
0.1 N Na2S2O3 till the contents become pale yellow in colour.

9 Add freshly prepared starch at the end of Nitration when the contents are light
yellow in colour.

37
Calculation

Volume of solution pipetted out = 10 ml

Volume of 0.1 N Na2S2O3 consumed for titration = A mL

1m L of 0.1 N Na2S2O3 = 0.0064 g Cu

A mL of Na2S2O3 = A × 0.0064 g Cu

This is present in 10 ml of the solution

Therefore in 1000 mL = A × 0.0064 ×1000

---------------------- g Cu
10

38
 1.11 Complexometry

(Estimation of calcium and magnesium by Versenate method)

Principle

EDTA (Ethylene Diamine Tetra Acetic Acid) forms stable com plexes with
various polyvalent cations at different pH levels. Calcium and magnesium gets
complexed by EDTA in the order of calcium first and magnesium afterwards at different
pH levels of 12 and 10 respectively. Hence calcium is first estimated by titration with
EDTA using murexide as indicator in the presence of NaOH at pH 12. Then magnesium
is estimated together with calcium using erichrome black –T indicator at pH 10 in the
presence of NH4Cl –NH4 OH buffer solution. Sn, Cu, Zn, Fe, Mn may interfere in the
determination of Ca and Mg if present in appreciable amount. There interference is
prevented by the use of 2% NaCN solution or carbomate. Usually in the irrigation water,
the quantities of interfering ions are negligible and can be neglected.

Apparatus required

9 Porcelain dish

9 10 mL pipette

9 25 mL burette

9 Glass rod

Reagents required

9 0.02 N EDTA solution

9 10 mL NaOH solution

9 Murexide indicator

9 NH4Cl –NH4 OH buffer

9 Erichrome black –T indicator

39
Procedure

9 Pipette out 10 ml of the given solution to a 250 ml conical flask.

Procedure

Calcium alone

9 Pipette out 25 mL water into a porcelain dish

9 Put a red litmus paper and add 10 mL of NaOH solution till the red litmus paper
turns into blue.

9 Then add a pinch of murexide indicator.

9 Titrate this against 0.02 N EDTA till the colour changes from pinkish red to
violet. Calcium + Magnesium Pipette out 25 mL water into a porcelain dish Put a
red litmus paper and add ammonium chloride-ammonium hydroxide buffer
solution till the litmus paper turns into blue Then add two drops of Eriochrome
black - T indicator Titrate this against 0.02 N EDTA solution till the colour
changes from purplish red to sky blue colour.

Calculation

Volume of irrigation water taken = 25 mL

Volume of 0.02N EDTA used for calcium alone = A mL

Volume of 0.02N EDTA used for calcium + magnesium = B mL

Volume of 0.02 N EDTA used for magnesium alone = (B-A) mL

Calcium

1mL of 0.02 N EDTA = 0.0004 g of Ca

A mL of 0.02 N EDTA = 0.0004 × A g of Ca

This is present in 25 mL of irrigation water =

106 mL = 0.0004 × A
-------------- × 106 ppm
25

40
 In terms of me L-1 = 0.0004 × A × 1000 1000
------------------------ × -----------
25 20

Magnesium

1mL of 0.02 N EDTA = 0.00024 g of Mg

Therefore (B-A) mL of 0.02 N EDTA = 0.00024 × (B-A) g of Mg

This is present in 25 mL of irrigation

water

106 mL = 0.00024 × (B-A)

-------------- × 106 ppm
25

In terms of me L-1 = 0.00024 × B-A × 1000 1000

----------------------------- × -----------
25 12

41
 1.12 Precipitometry (Estimation of chloride)

Principle

The chloride present in irrigation water is precipitated as silver chloride by

titration with standard silver nitrate solution using potassium chromate as indicator. After
all the chloride is precipitated, the excess of silver nitrate combines with potassium
chromate (indicator) to form flesh red precipitate of silver chromate which serves as the
end point.

Reaction

NaCl +AgNO3 AgCl + NaNO3

K2CrO4+ 2 AgNO3 Ag2CrO4+ 2 KNO3

Apparatus required

9 Porcelain dish

9 Glass rod

9 50 mL pipette

9 50 mL burette

Reagents required

9 0.1 N AgNO3

9 Potassium chromate indicator (5% aqueous solution)

Procedure

9 Pipette out 50 mL of the irrigation water into a porcelain dish.

9 Add a few drops of potassium chromate indicator.

9 Titrate against 0.1 N AgNO3 till the flesh red precipitate of silver chromate appears.

9 From the amount of 0.1 N AgNO3 consumed, calculate the chloride content.

42
Calculation

Volume of water taken = 50 mL

Volume of 0.1 N AgNO3 used = A mL

1 mL of 0.1 NAgNO3 = 0.00355 g of chloride

A mL of 0.1 N AgNO3 = 0.00355 x A g of chloride

This is present in 50 mL of water

In 106 mL = 0.00355 × A
-------------- × 106 ppm
50

In terms of me L-1 = 0.00355 × A × 1000 1000

-------------------------- × -----------
50 35.5

43
 1.13 Gravimetry (Estimation of Sulphate)

Principle

The sulphate in water is precipitated as barium sulphate by the addition of barium

chloride in hydrochloric acid medium. The precipitate is filtered, washed free of chloride,
ignited and weighed as barium sulphate.

Reaction

Na2SO4 + BaCl2 BaSO4+ 2 NaCI

BaSO4 + 2 C Filter BaS + 2 CO2
BaS + 2 O2 Atmospheric O2 BaSO4
Apparatus required

9 50 mL pipette

9 250 mL beaker

9 Silica crucible

9 Chemical balance

9 Water bath

9 Desiccators

Reagents required

9 Dilute hydrochloric acid (1:4)

9 10% barium chloride solution

9 Concentrated H2SO4

9 Concentrated HCI

9 Solid ammonium chloride

Procedure

9 Pipette out 50 mL of irrigation water into a clean 250 mL beaker

9 Add 10 mL of dilute HCI and one gram of solid ammonium chloride

44
 9 Heat to boiling and add 10 mL of 10% BaCl2 solution drop by drop with constant
stirring, continue boiling for another 2 to 3 minutes

9 Allow the precipitate to settle and test for completion of precipitation by adding a
small amount of BaCl2 solution through the sides of the beaker

9 If any turbidity is noticed, add sufficient quantity of barium chloride solution to

precipitate all the sulphate

9 Transfer to a hot sand bath and digest for half an hour to promote granulation of
the precipitate

9 Filter through Whatman No.42 filter paper and wash with boiling water till the
filtrate runs free of chloride (Test with silver nitrate solution)

9 Transfer the filter paper along with the precipitate to a weighed silica basin and
dry it in air oven Ignite over a low flame initially, taking care to ash the filter
paper completely. Then ignite strongly over a rose head flame to constant weight.

9 From the weight of barium sulphate obtained calculate the sulphate content of the
irrigation water.

Calculation

Volume of water sample pipetted out = 50 mL

Weight of empty silica crucible = Ag
Weight of crucible + BaSO4 (precipitate) = Bg
Weight of BaSO4 = (B-A) g
233.3 g of BaSO4 contains 96 g of SO4
Therefore (B-A) g of BaSO4 contain = 96 × (B-A)
---------------- g of SO4
233.3
This is present in 50 mL of irrigation water
In 106 mL = 96 (B-A)
-------- × ------- × 106 ppm
233.3 50
In terms of me L-1 = 96 (B-A) 106
-------- × ------- × ---------
233.3 50 48

45
 2. SOIL PHYSICS

2.1 Collection and Preparation of Soil Samples for Laboratory Analysis

Soil is analyzed to know the nature of soil, to classify and advocate to fanners
about the peculiarities of the soils and how much of fertilizer should be applied for better
crop production.

A complete soil, of course, cannot be moved into a laboratory. The value of the
laboratory work depends upon care in sampling. Each soil sample needs to be a fair
representative of the specific area or horizon worth sampling.

If the sample is to be a representative of an area, it is necessary to take large

number of samples spread over the area and then pool and sub-sample it so as to get a
sample of desired size. For soil survey work, samples are collected from a profile typical
of the soil of the surrounding area.

TOOLS

i) Spade, ii) Khurpi, iii) Auger (screw or post hole type), iv) Core sampler, v) Soil
testing tube (for wet soil), vi) Sampling bags and vii) Iron or plastic basin or bucket.

Collection of Soil Samples from the Field

Normally each field may be treated as a sampling unit. But two or more fields which
are similar in appearance, production and past-management practices may be grouped together
into a single sampling unit. Samples should be collected separately from areas which differ in
soil colour or past-management practices such as liming, fertilization, cropping pattern etc.
During collection of soil, avoid dead furrows, old manure or lime piles, wet spots, areas near
trees, manure pits, compost pits and irrigation channels. A wise collecting agent is one who
collects samples in the presence of the owner or cultivator of the land who is the best judge in
deciding which area of his farm should be sampled separately.

Scrap away the surface litter and insert the sampling auger to plough depth (15 cm).
Take at least 15 samples randomly distributed over each area and place them in a clean
bucket. If sampling auger is not available, make a ‘V’ shaped cut to a depth of 15 cm
using a mamutty or spade and remove 1,0 to 1.5 cm thick slice of soil from top to bottom
of the exposed face of the 'V shaped cut and place in a clean bucket or basin.
46
 Thoroughly mix soil samples taken from 15 or more spots of each area. By quartering,
discard all but ½ to 1 kg soil. Quartering is done by dividing the thoroughly mixed soil
into four equal parts and discarding two opposite quarters. Re-mix the remaining two
quarters and again divide into four equal parts and reject the opposite two. Repeat this
procedure until about ½ to 1 kg of soil is left. Put the soil into a clean, numbered, cloth bag.

Note :

9 Sampling should be done after the harvest of the crop.

9 In case sampling is necessary during crop growth, sample between the lines of
growing plants.

9 Avoid sending soil samples in fertilizer bags.

Collection of Soil Samples from a Profile

After the profile has been exposed clean one face of the pit carefully with a spade
and note the succession and depth of each horizon. Prick the surface with a knife or edge
of the spade to show up structure, colour and compactness. Describe the profile as per the
standard terminologies. Use the Munsell colour chart for noting the colour and find out
the texture by feel method.

From each horizon take sample by holding a large basin at the bottom limit of the
horizon while the soil above is loosened by a khurpi. The sample is mixed and transferred
to a bag after labelling.

Preparation of Soil Sample for Analysis

The soil sample received at the laboratory is air dried in shade and spread on a
sheet of paper after breaking large lumps if present, with a wooden mallet. It is further
ground by pounding with a wooden mallet in such a way that the aggregate particles are
broken down to ultimate soil particles. The soil thus prepared is sieved through a sieve
with round holes, 2 mm in diameter. The material on the sieve is again ground and sieved
till all aggregate particles are fine enough to pass through and only stones and organic
residues remain on the sieve. Mix well the 'fine soil' got by sieving and store in a suitable
bottle with one label on the outside and another inside the container.

47
Sub-Sampling for Analysis

The soil in the bottle is emptied on a clean thick sheet of paper and evenly spread
with a sampling knife. It is heaped into a cone by raising the four ends of the paper. It is
again mixed well and evenly spread on the paper as before. The process is repeated 3 or 4
times to ensure uniformity and then finally spread evenly on the paper again. Now it is
divided into four equal quarters and small quantity of soil is taken from various points in
each quarter to get a representative sample for the analysis.

Collection of Core Soil Samples

For the determination of bulk density, hydraulic conductivity experiments

undisturbed soil core should be taken. Earlier, cups were used to determine the hydraulic
conductivity and bulk density. The cups have two cylinders lower cylinder with threads
on inner side and upper one with a nozzle. The disadvantageous in this method are,

1. Volume is very much less.

2. Because of hammering compaction problem may arise.

Now a day's core sampler is used for this purpose. It consists of.

1) Core-cutter:

It has got a cutting sharp edge. The cutting edge is of diameter 7 cm which
coincides with diameter of the cylinder. The length of the cutter above the edge is about
10 cm (sometimes 11.5 cm). Upper edge of the cutter has a screwing-thread to a length of
2.5 cm.

2) Sectional Cyclinder:

Diameter and length are 7 cm. sometimes the sectional cylinder length is of
8.5 cm. it has got guard rings of same diameter with 1.5 cm depth.

3) Collar:

The diameter of the collar is 10.5 to 11 cm. so that it exactly fixes into the core
cutter. Collar has an opening with 3 mm diameter on one side so as to release air. It has a
solid neck over which a hollow cylindrical handle is attached.

48
4) Hammer:

Cylindrical iron bar having a handle on the upper side and a grinding rod attached
to the lower side of the hammer.

Precaution:

9 The area to be sampled should be cleared off from vegetation.

9 If there are lots of cracks, reject that place for sampling. Especially black soil.

9 If the area is very dry and loose, we can moister the area with little amount of water.

9 If the land is too wet, allow the soil to dry then take sampling.

Procedure:

After thorough cleaning, the sectional cylinder with guard rings were placed on
both sides insides the core cutter. Till the lower surface of the collar touches the soil, it
was hit inside the soil. In vary loose soils, the collar should be hit still more. After
scooping the soil ground core cutter, the cutting unit was separated from the handle. The
soil from the cutting edge was scooped, the soil core was pushed gently so that the
sectional cylinder holding the soil will core out care should be taken to see that the
position is not changed.

After removing the samples, the excess soil was shaved off from the guard rings.
Labeling should be done properly indicating the upper surface of the sample. Both sides
of the ring were covered with muslin cloth so that sample will not fall while transporting.
Transporting should be done with the help of special boxes made with the help of special
boxes made with capacity to hold 12 rings.

After reaching the lab, the guard rings were removed immediately and placed in
water. Samples with pebbles should be rejected. In the laboratory samples should be
placed in a tray containing phenol to avoid (wash) the growth of algae.

49
 2.2 Determination of Bulk Density, Particle Density and Pore Space

An important basic parameter for all soils is the relative amount of pore space and
solid space. The bulk density (mass of dry soil divided by its undisturbed volume) is the
central property used to determine soil mass and porosity.

Bulk density is the mass per unit volume of soil including pore space. The normal
range of bulk density in most soils is from 1.02 to 1.80 Mg / m3. The bulk density of
coarse textured soils will be on the higher side while organic soils will have lower bulk
density. Porosity is that fraction of soil volume not occupied by solid particles. Particle
density is the mass per unit volume of soil excluding pore space. Soil bulk density (mass
of dry undisturbed soil per unit volume)

Soil Bulk Density

Bulk density is sometimes estimated based on previous evaluations or directly

determined by field and laboratory analytical methods. Soil bulk density remains constant
over hundreds of years unless disturbed by anthropogenic activity such as heavy
equipment, tillage compaction, or excavations. Anthropologic disturbances are often
drastic and permanent. Bulk density increases with increasing soil depth for undisturbed
("natural") soils. Values for bulk density are usually given in grams per cubic centimeter
or mega grams per cubic meter (g/cm - Mg/m ). Rock usually contains a small amount of
pore space compared to soil. Rock bulk densities are commonly greater than 2.0g/cm. A
solid cubic centimeter of the mineral quartz is 2.65 g/cm (specific gravity).

Aim: To determine the bulk density, particle density, and pore space in the given soil
sample

i) Cylinder method

Principle

The bulk density and percent pore space are determined from the apparent and
true volumes of the soil measured by adding a known quantity of water to a measuring
cylinder containing a weighed quantity of soil.

50
Apparatus required

a) Measuring cylinder (100 mL) with glass stopper

b) Chemical balance or Electronic balance

Procedure

Weigh exactly 20 g of soil sample and transfer in small quantities at a time to a

100 mL measuring cylinder with glass stopper by gently tapping the cylinder. After
completely transferring the soil, note the volume. Add a known volume of water (50 mL)
along the sides of the cylinder using pipette till the entire soil mass is completely soaked.
There should be at least 5 mL of water above the soil surface after the addition of water.
Care should be taken to see that the final volume of soil and water is below 100 mL mark.

Keep the cylinder with soil and water in an undisturbed condition for at least 30
minutes so that the entire pore space is completely filled with water. Note the final
volume of soil plus water after the expiry of time and calculate the percentage of bulk
density, particle density and pore space.

Calculation

Weight of the soil taken = W (20) g

Volume of soil taken = V1 mL
Volume of water added = V2 mL
Volume of soil + water = V1+V2 mL
Volume of soil + water at the end of = V3mL
experiment
Pore Space volume = (V1 + V2)-V3 mL
Percent Pore Space = (V1 + V2)-V3 mL
----------------------- × 100
V1
Bulk Density (W/V1) = Weight of soil
------------------- Mg/m3
Volume of soil (V1)
Particle Density = W
----------------------- Mg/m3
V1 - Pore space volume

51
ii) Wax coating method

Principle

A soil clod of known weight is coated with wax and immersed in a known volume
of water and from the volume thus arrived at, the bulk density is determined.

Apparatus required

1. Balance

2. 250 mL measuring cylinder

3. Twine

Reagents required

1. Wax

Procedure

Take a small clod from the field, the bulk density of which is to be determined
and note its weight. Then tie the .clod with a twine. Immerse the clod in melted wax so
that there is a complete coating of the clod with wax. Take a known volume of water in
a250 mL measuring cylinder. Immerse the wax-coated clod in water and note the rise in
volume of water in the cylinder. Calculate the bulk density of the clod from the weight
and volume of the clod.

Calculation

Weight of the clod = W g

Initial water level in the cylinder = A mL

Final water level in the cylinder after = B mL

Immersing the clod

Volume of the clod = (B - A) mL

Bulk density of the soil = W

--------- Mg/ m3
(B-A)

52
 2.3 Determination of infiltration of soil

(Double Ring Infiltrometer Method)

When water is applied at the soil surface as irrigation or is received as rainfall it

enters into the soil profile and moves laterally and down wards. The process of entry of
water into the soil refers to infiltration. The maximum possible infiltration rate is called
the infiltration capacity or infiltrability of the soil. Quantitatively infiltrations rate is the
volume of water passing into the soil per unit area per unit time and has the dimensions
of velocity (LT~]). Infiltration determines the amount of water that enters soil and that
water flows over the soil surface is called surface run off-over land flow. Infiltration rate
is influenced by soil characteristics like texture, structure, total porosity, hydraulic
conductivity, initial water content, impeding layer and organic matter, tillage and
cropping practices and rainfall intensity.

Infiltrometer method:

Principle

When water is ponded inside concentric metallic rings driven into the soil. The
rate of recession (fall) of the water level of the rate of water withdrawn from a supply
source used to maintain a constant head of water on the soil surface gives the infiltration
rate of the soil. During infiltration of water into the soil appreciable lateral movements of
water may also occur. To avoid the error due to this lateral movement two concentric iron
rings are used. Equal's heads of water are maintained in both the rings. The infiltration is
measured in the inner ring only. The rate if intake per unit area varies markedly with the
size of the rings and the depth of soil to which they are driven into. Infiltration rate
studies are preceded by a careful description of texture, structure, surface conditions,
layering sequence in the profile, initial soil moisture and any other specific problems of
the regime in which the soil is located.

Apparatus

1. Cylinders are made of 14 to 16 gauge iron sheet rolled in circular fashion and
joints are ground to a smooth; fin. One end of the cylinders is sharpened from
outside keeping the inside completely smooth. This facilitates easy drive of the

53
 cylinders in the soil. Diameter of the inner cylinder is generally 30-35cm and that
of outer cylinder 40-45 cm. The height is generally 40-145 cm.

2. Circular caps or plates made of iron 1-1.5 cm thick iron sheet and 5-10 cm larger
in diameter than the diameter of the largest cylinder.

3. Peizometer: Triangular right angular wooden stand with the slope of 1:2 (IHeighr
and 2 Slope) to avoid parallax error. Where in a graduated glass tube is mounted
/ on the scale of hypotenuse.

4. Hammer of sufficient weight to drive the rings into the soil. w

5. Hook gauge or staff gauge.

6. Field source of water

7. Thermometer

8. Watch

9. Rubber or plastic sheet to be used as splash guard

Procedure

x Select a representative area in the field.

x Examine its texture, structure and water content.

x Record conditions of soil surface such as surface litter Or mulch, uncultivated with
i crust and/or cracked surface or freshly cultivated with friable cloddy surface etc.,

x Set the inner cylinder in place in the selected area and drive it vertically
downwards into the soil with the help of driving plate and hammer to a desired
depth (Generally 10-20cm). .

x Care should be taken that the soil is disturbed to a minimum and cylinder is
driven straight downwards from all sides. There should not be any crack.

x Tap the soil into the space between the soil column and the inner side of the
cylinder to its natural condition as far as practicable.

x Set the outer cylinder concentric to the inner one and drive it to the same depth
taking the same precautions as taken in the case of inner cylinder.

x Cover the soil with splash gaurd

54
 x Apply 10-15 cm water in the central (inner) ring as will in the space between the
central and outer ring almost simultaneously.

x Remove the splash guard.

x Measure the level of water in the central cylinder with the gauge keeping edge of
the cylinder as reference level.

x Record the recession of water in the inner cylinder at 1, 3, 5, 10, 20, 30, 40, 60,
80, 100, 120 minutes and hourly thereafter till the water intake is almost constant.
However, the time intervals of observation can be carried according to the
objectives of the study and the soil permeability. If all the ponded water soaks
into the soil in a short period, more water may be ponded and the recession of
water recorded again and again till the constant intake rate is reached.

x Record the water level and time just before and after responding. Keep the intervals
between these two observations as short as possible to avoid errors caused by
intake during the refilling period.

Precautions

1. Install inflltrometer straight down in order to minimize soil surface disturbance

2. Cover the soil surface in the rings with polyethylene sheet (splash guard) while
pouring water in the rings.

3. Try to keep water level identical in the inner as well as outer ring.

4. If you are working in summer when evaporation rate is high cover the water
surface with oil film to minimize error due to evaporation.

5. The temperature of the infiltration water should be the same as that of the soil to
avoid soil air being dissolved in the infiltration water thus increasing infiltration
rate. This happens when temperature of the water is more than the soil. Reverse
will be the case if infiltration water is colder than the soil.

6. Water used in inflltrometer tests mast be of same quality and composition as the
water in the real system for which the infiltration rate is to be determined.

55
Factors affecting infiltration

a) Precipitation

b) Soil characteristics

c) Land cover

d) Slope of the land

e) Evapotranspiration

Soil type Steady state infiltration rate (mm/hour)

Sandy 30

Loamy sand 20-50

Sandy loam 12-20

Silt loam 7-12

Clay loam 3-7

Clayey <3

Classification of infiltration rate

Rating Infiltration cm hr -1

Very rapid 25.4

Rapid 12.7-25.4

Moderately rapid 6.3-12.7

Moderate 2.0-6.3

Moderately slow 0.5-2.0

Slow 0.1-0.5

Very slow <0.1

56
Observation and calculations:

Record the following information

1. Location

2. Texture of the surface soil

3. Surface conditions(Crop residues, Litter, clods, cracks, crust, salts, kind and
extent of vegetation)

4. Layering sequence in the profile (If profile has been studied)

5. Specific problem, if any (erosion, salinity, ponding)

6. Temperature of the water

Infiltration rate P1-P2

------------- × 60
2 (t2-t1)

Where,

P1 & P2 = Initial and Final peizometer readings

t 1 -t2 = Initial and Final time

2 = Conversion factor for peizometer readings

Piezometer Reading Actual Drop

Cumulative Actual
of water level Infiltration
S.No Time Time
Final Rate (cm hr -1)
(Minutes) Initial (P1) (P2 - P1) (Minutes)
(P2)

57
 2.4 Determination of Aggregate Size Distribution of Soil

Under natural conditions primary soil particles particularly clays tend to cohere to
form secondary units called aggregates. Analysis of aggregates refers to determination of
size distribution of aggregates. The size distribution of aggregates in soils indicates their
susceptibility to erosion, movement and retention of water and air because their size is
important in determining the dimensions of pore space in cultivated soils. The extent of
disruption of aggregates due to rains or irrigation depends on the water stability of
aggregates. Crop yields are significantly correlated with aggregates of soil are equal.
Larger inter-aggregate pores favour high infiltration rate and adequate aeration for plant
growth. The well aggregated soil are resistant to all types of erosion and provide free
drainage with adequate retention of water. Aggregate analysis can be carried out with dry
soil sample or by wet sieving depending upon purpose.

Wet sieving method

The size of water stable aggregates is greatly affected by the method of wetting of
the soil sample. Direct immersion of dry soil in water at atmospheric air pressure causes
disruption of larger aggregates into smaller aggregates. This disruption can be
considerably reduced by wetting soil sample slowly under vacuum, under tension or by
'spraying with fine mist of water. The choice of method depends on the objective of the
analysis, where the objective of the work is to investigate the resistance of soil to erosion
by water or formation of surface crusts, wetting the soil by direct immersion is preferred.

Principle

A known amount of the soil sample collected from the field is immersed in the
water for a short period to stimulate wetting rains or flood irrigation. The wetted sample
is sieved through a nest of sieves in Yoders apparatus, which raises and lowers the nest of
sieves. This is done to stimulate the disruptive forces of water and facilitate the sieving of
water stable aggregates through sieves. After 30 minutes sieving the sieves are placed in
oven at 105°C to dry the aggregates to a constant weight. The dry aggregates are
collected and weighed. These aggregate weights also include weights of primary particles
of primary particles of respective sieve sizes. Therefore, soil of these aggregates are
dispersed and passed through the respective sieves. The mass retained on respective

58
sieves represents primary particles or sand fraction and hence is deducted from the mass
of aggregates to get correct estimate of aggregates. A graph plotted for aggregate mass
against aggregate size gives the size disruption of aggregates. To represent the aggregation
status of soil by single value following indices can be calculated.

MWD = ෍ †‹ ‫‹ כ‬
௜ୀଵ

Where MWD is the mean weight diameter of aggregate, n is number of size fractions
including the finest that passes through the finest sieve, di is the mean diameter of each
size fraction; wi is the weight of aggregates occurring in the corresponding size fraction.

෍ ‹ ‫‹† ‰‘Ž כ‬
௜ୀଵ

GMWD = ----------------
௡

෍ ‹
௜ୀଵ

Where, GMD is the geometric mean diameter.

Apparatus:

1. Nests of sieves 20.0 cm in diameter and 5 cm in height, with mesh numbers 4, 8,

16, 32, 64 and 160 (i.e. hole width of 4.00, 2.00, 1.00, 0.5, 0.25 and 0.10 mm,
respectively).

2. One 2 mesh screen ( hole width of 8 mm )

3. A Yoder (1936) type sieving machine which raises and lowers the nests of sieves
3.75 cm through water 30 times per minute.

4. Mechanical stirrer with a bowl

5. Moisture boxes

6. N/ lO NaOH

59
Procedure

¾ Take the sample when the soil is moist and prepare the sample by gently
breaking the clods so as to pass through 8 mm (2 mesh) and retained on 4mm
(4-mesh) screen.

¾ Air dry the sample collected over 4-mesh sieve, weigh three representative 25 g
sub-samples. Oven dry and weigh one sample. Use the two samples not oven-
dried as duplicates in following determinations.

¾ Mount the sieves upon the racks and suspend these in the water container tank.
Raise the racks to maximum height.

¾ Fill the container with salt free (EC < 10~5 dS m"1) water to a level little below
that of the screen in top.

¾ Place the air dry sample evenly over the top sieve.

¾ Raise the water level rapidly or lower the racks rapidly to cover the sample with
water.

¾ After 10 minutes of wetting the sample, begin shaking or sieving underwater for
30 minutes.

¾ Remove the nests of sieves from the water tank, drain them in an inclined
position and put in an oven for drying at low temperature for an hour.

¾ Transfer the contents of sieves to pre-weighed aluminum boxes or glass

beakers, dry them to a constant weight at 105°C and determine the weight of
aggregates.

¾ Transfer the oven dry soil aggregates for all moisture boxes of a given set into
dispersion bowl. Add dispersing agent (10 ml of 5% solution of sodium hexa
meta phosphate for Ca++saturated soil or 4% solution (1 N) of NaOH for
acid soils) and enough distilled water to fill the bowl within 10 cm or rim; then
stir the suspension for 10 minutes.

¾ Wash the suspension on an identical set of sieves by means of a stream of tap

water.

60
 ¾ Sand remained on each sieve is oven dried and weighed.

¾ Calculate material smaller than 0,21 mm by subtracting the sum of oven dry
weight of original sample.

¾ Correct the moist weight of original sample to dry weight of original sample by

¾ Knowing moisture content from third sub-sample.

Observations:

1. Mass of fresh sub samples = 25 g

2. Mass of oven-dry sub sample = Wtd g

3. Masses of aggregates collected from sieves

Masses of aggregates collected from different sieves

Mass of water stable

Mass of aggregates + Sand Mass of sand
Sieve size aggregates
(Wti, g) (Ws, g)
Wji = Wti-Ws g
>2.0
1.0
0.5
0.25
0.1
<0.1
Calculations

1. Convert corrected weight of water stable aggregate to percentage in each size

class as follows

Wji
----- × 100
Wtd

Where ,Wj is the mass of corresponding aggregate size class (i)

2. Calculate MWD and GMD by putting the values of 'di* and wi in equations (i) and (ii)

61
 2.5 Estimation of Soil Water Potentials Using Tensiometer

A tensiometer is a moisture measurement tool that measures how hard the plant is
working to extract water from the soil. It directly measures the physical force that the root
system must overcome in order to access water held in the soil (also known as matric
potential). Tensiometers read in kilopascals (kPa). The higher the reading on the gauge,
the harder it is for the plant to extract water. A working tensiometer consists of a sealed
tube filled with water, a porous ceramic tip and a vacuum gauge. Tensiometers can be
used to predict crop water demand and therefore irrigation requirement,

Principle

Tensiometers operate by allowing the soil solution to come into equlibrium with a
reference pressure indicator through a permeable ceramic cup placed in contact with the
soil. The tensiometer is inserted into the soil so that the ceramic tip is at the depth of soil
that is to be monitored. The ceramic tip of the tensiometer must be in contact with the soil
in the active root zone in order to obtain useful information. As the soil dries out, water is
sucked out of the tensiometer through the porous ceramic tip. This creates a partial
vacuum in the tube, which registers on the vacuum gauge. Conversely, if irrigation or
rainfall occurs and the soil around the ceramic tip becomes wet, water is sucked into the
tube and the vacuum is reduced. Readings should be taken at least 2-3 times per week
during summer.

Reading Interpretation*

0-8 Soil is very wet. Reduced crop growth will result.

8 Field capacity

8-35 Best conditions of soil moisture for crop growth

35-50 Mild stress on well-drained soils

50+ Soil is very dry. Yields will be affected

* Readings may vary depending on soil type.

62
Disadvantages

a) Point measurement (in as much as there is no model capable of integrating a

larger soil volume such as is possible with some sensors).

b) Slow reaction time.

c) Source of water during soil drying.

d) Usually only operate between saturation and about -70kPa (They are thus not
much use for measurements in the dry end of the spectrum.) Air bubbles may
enter at this point (termed the Air entry potential). If the soil is very coarse, this
occurs even sooner (wetter).

e) Require maintenance to refill after dry periods.

f) If the ceramic cup looses contact with the soil (in an air pocket created by
manuring for example), then this could cause an apparent "lack of response" of
the instrument. If the tip were in an area of limited root activity, the readings
could also "stall". Air will enter a standard tensiometer certainly near the limit of
its measurement--75-85 kPa. I have not kept up on this discussion, but I do recall
a good bit of amendment with manure which may have served to create air
pockets in the soil. If the tip of the instrument were to be located in one of these
air pockets then contacts between soil and tip would be lost.

Advantages

a. They are not affected by the osmotic potential of the soil solution (the amount of
salts dissolved in the soil water), as the salts can move into and out of the ceramic
cup unhindered. This is not to say that the plant does not feel the effect of the
osmotic potential.

b. They measure soil matric potential with good accuracy in the wet range. Suited to
applications where water stress and irrigation needs have to be monitored. Less
subject to localized spatial variability than volumetric water content measurements
(and therefore less sensitive to soil disturbance during installation).

63
 2.6 Determination of Field Capacity of Soil

Veihmeyer and Hendrickson (1931) defined field capacity as the amount of water
retained in a soil, following saturation, after downward water movement has materially
ceased. This definition implies that field capacity is a point of static equilibrium. But
static equilibrium is never reached under field conditions. In the absence of evaporation
downward movement of water continues for a longer period (Richard et al., 1956).
Although the concept of field capacity has been widely criticized on the basis of theory of
water movement, yet practical applicability of this concept as the upper limit of water
availability cannot be ruled out. Quite generally field capacity has been equated with
water retention at 0.1 bar suction in coarse textured and 0.33 bar suction in medium and
fine textured soils, respectively. But critical studies have shown that field capacity is
essentially a field value and needs to be evaluated in the field as in some soils field
measured values differs from laboratory values as much as 75 per cent.

Apparatus

1. Tensiometer assembly

2. Access tubes

3. Neutron probe assembly

4. Spade

5. Polyethylene sheet

6. Straw

7. Water reservoir

Procedure

• Enclose is a small area (2m×2m) in the field by earthen dikes.

• In the central portion of the area, install 2-3 access pipes up to the desired depth for
measurement of soil moisture with neutron probe.

• Similarly place 3-4 tensiometer at this depth around the pipes for recording
moisture tension.

• Pond the diked area with water till the profile is saturated to the desired depth.

64
 • As soon as the ponded water is infiltrated cover the area with polyethyelene film
and straw so as to check evaporation and temperature changes at the surface.

• Monitor volumetric soil water content.

• Calculate the total soil water storage in the profile over short intervals till it
becomes nearly constant,

• Monitor the soil water suction from tensiometers also.

• This mean water storage is a reliable estimate of field capacity of soil at the tension
monitored above.

Observation and calculation

Volume of water Storage

Depth (cm) Soil thickness (m)
content (m3) (m)

Results:

65
 2.7 Determination of Saturated Hydraulic Conductivity of Soil

The saturated hydraulic conductivity of soil refers to the readiness with which a
saturated soil transmits water through its pores and is expressed as length per unit time.
It depends upon several soil factors and properties of the fluid. The soil factors affecting
the hydraulic conductivity are i) pore geometry, ii) mineralogical composition of the soil,
iii) stratification, iv) presence of entrapped air in the soil pores and v) organic matter or
the microbial activity in the soil. Since viscosity and density of the fluid passing through
the soil affect the hydraulic conductivity this soil property varies for different fluids.
If saturated hydraulic conductivity is adjusted to account for viscosity and density of the
fluid. We obtain the intrinsic permeability which depends only upon the soil properties.
Thus, the intrinsic permeability of a stable soil is same for all fluids. The saturated
hydraulic conductivity (Ks) is related to the intrinsic permeability (K') as follows

Ks = K' dg / ŋ

where d is density (Mg m-3), Ŋ is the viscosity (Kg m-2 s-1) of the fluid and g is the
acceleration due to gravity (ms-2). The solid liquid interaction if any can influence Ks
through its effect on the nature of the pores and the overall porosity. The hydraulic
conductivity of soils may vary from 0.001 cm hr"1 in fine clays to over 25.0 cm hr"1 in
coarse sand and gravel.

Methods of determination Constant water head method Principle

If the constant water head is maintained on one end of saturated column of soil of
length L. the volume of water (Q) percolating through the other end per unit cross-
sectional area, A, of the soil column per unit time (t) will be directly proportional to the
hydraulic gradient (ΔH/L) over the length of the soil. Thus

Q/At = Ks ΔH/L

according to darcy's law the proportionality constant Ks in the above equation is the
hydraulic conductivity of the soil. The symbol H in equation stands for the total head
which is the sum of the hydraulic head (h) and the gravity head (z) at any point in the soil
column. The difference of H at the top and bottom of soil column divided by the length of
the column gives the hydraulic or the total head gradient.

66
PRINCIPLE

The rate of flow of a liquid through a porous medium depends on the size, distribution
and continuity of pores and the temperature of the fluid which is indexed as hydraulic
conductivity. In case of soil saturated with water, it is directly related to the permeability of the
porous medium. Using constant head of water, water is passed through the soil column and the
conductivity of water per unit time is calculated using Darcy's equation

Ks (or) K sat = QL / At ΔH

(Ks expressed in cm hr-1)

Where Ks - Saturated hydraulic conductivity

Q- Quantity of water flows out at a time interval of t" in ml

L- Length of hydraulic head in cm

A- Cross-sectional area of the soil column in cm2

ΔH- water head or hydraulic head (L + column of water standing over the soil) t—time in
hour

Apparatus

• Hydraulic rings

• Measuring cylinder

• Stop clock

• A water reservoir with arrangement for maintaining a constant head of water over the
soil column.

Soil-water saturated flow follows an equation proposed by Henri Darcy a French

engineer (1856). This equation relates the quantity of water (Q) moving through a soil to
the hydraulic gradient (H), the length of the flow path (L), the cross sectional area (A),
time period (T), and the hydraulic conductivity of a soil layer (K), as follows:

H
Q = KAT -----
L

67
Where:

Q = gallons/hour (1 gallon = 231 cubic inches)

K* = inches/hour (a constant for each specific layer)

A = inches squared (area circle = πr2)

T = hours

H = height of water in inches above a reference level

L = length of flow path in inches

K is the soil parameter, which can be estimated by observing and determining soil
particle size distribution (texture), structure, and bulk density.

QL
Hydraulic Conductivity ks (or) ksat = ----------- cm hr-1
At ΔH

Where,

Ks — Saturated hydraulic conductivity

Q - Quantity of water flows out at a time interval of 't' in mL ( 7.0)

L - Length of hydraulic head in cm

A - Cross sectional area of the (Core) soil column in cm ( nr ) ( 38.5)

ΔH - Hydraulic head (L +column of water standing over the soil) (7.0+1.5 = 8.5)

T - Time interval

Procedure

• Collect Core samples.

• Keep it in a tray containing water overnight for saturation.

• Place a filter paper on the surface of the soil column to avoid the removal soil
particles by the inletting water.

68
 • Connect the hydraulic rings over saturated sample by means of a wax coating or
rubber bands.

• Place the sample on a metal screen supported with an out flow unit.

• Ensure the water supply into the constant head water supply unit and the
discharge tube on constant head was connected to hydraulic rings.

• Allow the water to flow over the surface of the soil column.

• Adjust the inflow and outflow of water.

• Allow 15-20 minutes for stabilization of flow.

• Note down the time by your watch when the water head on the soil sample
becomes constant and a steady flow is obtained at the bottom.

• Measure the water that is passing through the soil column in a stipulated time. 5
minutes for sandy soil and 15-20 minutes intervals for clay soils.

• Start collecting the percolate in a graduated cylinder or a beaker.

• Measure the volume of percolate collected in a known interval of time.

• Record a few consecutive readings till the flux is constant.

• Measure the exact water head on the soil and then discontinue the experiment.

• Measure the length of soil column.

• Note down the temperature of the water used in the experiment.

Permeability of the soil is calculated from the hydraulic conductivity using the relationship.

dgk
K = -----
ŋ
Where,

d - Density of the fluid.

g - Gravitational acceleration.

k - Hydraulic conductivity.

ŋ - Viscosity of the fluid.

69
Hydraulic conductivity rating

Rating Ks cm hr -1

Very slow < 0.125

Slow 0.125-0.500

Moderately slow 0.500-2.000

Moderately 2.000-6.250

Moderately rapid 6.250-12.500

Rapid 12.500-25.000

Very rapid > 25.000

Observations and calculations

Diameter of the core d = Cm

Radius of the core = Cm

Cross-sectional area of the soil column A = --- cm2

(πr2)

Depth of water above the soil, h = --- cm

Length of soil column, L = ---- cm

Time for which percolate collected, t = ---- min

Volume of percolate collected, Q = ----min

Hydraulic gradient (head) (L+h) = ---- cm

Soil Hydraulic conductivity (Ks) = QL

-------
At ΔH

II. Falling head method

Principle

In this method drop in water level in a narrow tube is measured instead of flow.
This method is adopted for slowly permeable soils.

70
 Suppose in a small time dt there is drop of water in a narrow tube of cross-sectional
area (a) inverted over a saturated soil, whose cross-sectional area (A) and length (L)

K = (aL/At) ln (h1/h2)
Apparatus

Special apparatus consists of a galvanized iron cylinder (40 cm in length, 30 cm

diameter) with a conical top to which is attached a vertical glass tube of small diameter.

Procedure

• Push the cylinder into the soil to a depth for which determination is to be made
and fit the whole apparatus.

• Wet the sample from the below by supplying water to the bottom of the sample
through a three way stop cock and lower porous plate.

• Fill the space above the sample up to the overflow with water. This may be done
by flow upward through the sample or by introducing water with pipette or
syringe at the top of the sample.

• Establish a water level in the stand pipe somewhat above the level by introducing
water through the three way cock.

• Connect the stand pipe to the sample by opening the stop cock and measure the

• Make additional measurement by repeating step 3 and 4.

Observations and calculations

Diameter of stand pipe = d cm

Area of the stand pipe = a cm
Length of the sample = L cm
Diameter of the sample = d cm
Cross-sectional area of the sample = πD2/4, A cm2
Initial hydraulic head = h 1 cm
Final hydraulic head = h 2 cm
Time taken for change in head = ts
Hydraulic conductivity = cm s

71
 Ks (cm hr'1)
Sr. No. Soil type
Constant head Falling head

Results:

72
 2.8 Determination of the Pore Space of the Soil

(Buckner Funnel Method)

Principle

The total volume of the porous medium composed of solid volume and void or
pore volume. When the soil is saturated with water, it attains "zero tension". At this stage,
the volume of water equal to total pore space of the medium. The difference in the
amount of moisture at the zero tension and at the given tension (50 cm water column)
represents the volume of macrospores. Similarly the difference in amount of moisture at
the given tension and at oven drying represents the microspores.

Procedure

The saturated core samples were weighed and kept on the Buchner funnel of the
porosity apparatus. A suction pressure was created with the help of 50 cm water column.
Care should be taken to avoid air bubbles in the apparatus. The soil samples were
removed as soon as the water drop cease in the outlet and weighed again. The loss of
weight was used in calculating macro porespace. Then the samples were oven dried and
weighed. The difference in weight from that of 50 cm water column suction was used in
calculating microspores.

Calculation

Non- capillary pore space (Volume of water at zero tension - Volume of water at
=
(%) 50 cm water column tension) / Volume of the soil * 100
OR
(weight of soil at zero tension - weight of soil at 50 cm
water column tension)
=
----------------------------------------------------------- × 100
Volume of the soil
(weight of soil at 50 cm water column tension - weight of
over dried soil)
Capillary pore space (%) =
-------------------------------------------------------------× 100
Volume of the soil
Total pore space (%) = Non-capillary pore space + Capillary pore space
Results:

73
 2.9 Determination of Moisture Content Under Different

Suction Levels (Pressure Plate Method)

Principle

The probability of a pore to retain moisture under different pressure depends upon
the size. When saturated soil is subjected to a pressure greater than zero tension, some of
the water will be drained away until the pores have a diameter corresponding to the
applied pressure.

Procedure

The rubber rings were placed on the ceramic plates and were filled with soil
sieved through 2 mm sieve. The porous plate was then kept in tray containing water such
that the rings immersed partly, and the samples were allowed for overnight saturation.
After saturation, the excess water was drained and kept inside the extractor and was
connected to the outlet stem of the pressure plate. The extractor was then closed with a
heavy lid. The pressure was regulated to have desired pressure (0.33, 1, 2, 3, 6, 9,12 and
IS bars). The soil sample was allowed to attain equilibrium to that of corresponding
pressure. Then, excess pressure was released and the extractor was opened. The pressure
plate was removed and the soil was transferred to the pre weighed aluminum containers
and the moisture content was determined gravimetrically.

Observation and calculation

Weight of soil + Aluminum of cane (0.33 bar) = a (g)

Oven dry weight of soil + Aluminum of cane (0.33 bar) = (b) g

Weight Aluminum of cane alone (0.33 bar) = (c) g

Moisture content at 0.33 bar = (a-b) = d g

Weight of dry soil 0.33 bar = (b-c)=e g

% of moisture at 0.33 bar (Field capacity) = (d/e) × 100

Weight of soil + Aluminum of cane (15 bar) = (A) g

Oven dry weight of soil + Aluminum of cane (15 bar) = (B) g

74
Weight Aluminum of cane alone (15 bar) = (C) g

Moisture content at 1 5 bar = (A-B)=D g

Weight of dry soil 15 bar = (B-C)=E g

% of moisture at 15 bar (Permanent wilting point) = (D/E) × 100

Available water = (Field capacity moisture - Permanent wilting point moisture)

Result

SI. No. Different Suction Level % of moisture

1. 0.33 (Field capacity)

2. 1

3. 2

4. 3

5. 6

6. 9

7. 12

8. 15 (Permanent wilting point)

9. Available water

75
 2.10 Measurement of Soil Temperature

Soil temperature refers to the intensity of heat expressed as degree. It is an

important edaphic factor that controls chemical and biological reactions in soil and
influences plant growth. The main processes influenced by soil temperature are,

• Seed germination, root and shoot growth.

• Absorption and transport of water and nutrients by plants.

• Decomposition of organic matter.

• Microbial and enzymatic activities.

• Weathering processes.

Different plants require different temperature range for their optimum growth and yield
e.g. maize 25-30°C, rice 25-35°C, wheat 15-25 °C, tomato 15-21 °C etc.

Soil temperature can be estimated by its influence on some property of matter that
responds to variation in intensity of heat in body of the matter. Changes in properties of
matter given in table 1 have been found useful for temperature measurements. There are a
number of thermometers that can be used for measuring soil temperature e.g. mercury of
liquid in glass, bimetallic, bourbon and electric resistance thermometers.

Thermal properties of different substances used for measuring temperature

Thermo-metric substance Thermo-metric property

Mercury or liquid in glass capillary Volume

Platinum or other wires Electric resistance

Thermocouples Thermal electromotive force

Gas or vapour at constant volume Pressure

Thermocouple method

The precise measurement of soil temperature can be made with thermocouples

which are made by joining two dissimilar metal at two different places to form two
junctions. If entire circuit is composed of only two metals, the total emf in the circuit is

76
proportional to the difference in temperature of the two junctions. One junction is called
measuring junction (hot) and the other is the reference junction (cold). To measure
temperature the reference junction is kept at constant temperature at 0°C (with melting
ice). However, some commercially available potentiometer have a built in correction for
the temperature of the reference junction.

The advantage of the thermocouples for measuring soil temperature in addition to

adaptability to automatic recording is their small size and almost instaneous response to
temperature change. They have low heat capacity and lead wires may be very small so
there is very little influence of the ambient temperature on reading.

Principle

When two dissimilar metals are joined at two different places forming two
junctions in a circuit, total emf in the circuit is proportional to the difference in
temperature of two junctions.

Apparatus

1) Copper wire (20-30 gauge)

2) Constantan wire (20-30 gauge)

3) Sleeves

4) Material for soldering (gun, wax, metal)

5) Potensiometer

6) Hot bath thermometer

7) Stirrer

Procedure

I. Fabrication of thermocouples

To fabricate the thermocouples two unlike metals are used, but there are some
metals like copper, ferrous, platinum, radium and iridium and some alloys like alume (2%
aluminium, 90% nickel and 8% manganese and silicon ), chromel (10% chromium and
90% nickel) and constant (60% copper and 40% nickel) are giving good results. For

77
agricultural purposes, copper- constantan and iron -constantan are mostly used. The size
of wire depends upon the temperature range in which one is interested to work. For high
temperature range large and thick and for low temperature range small and thin wires are
used. Thermocouples are fabricated as follows

• Twist the two wires with long nose pliers to create a junction.

• Solder the two wires together. Care must be taken that soldered junction is small
as it will take small time for response.

• Repeat the procedure for the second junction.

• Insulate the wires properly with sleeves.

II. Calibration of thermocouple

The output recorded by potentiometer gives the difference of temperature of the

two junctions and is 40 micro volts for each degree Celsius for copper-constantan
thermocouple. Calibration is done by measuring emf with water in the hot water bath at
different temperatures and calibration curve is made for emf and temperature.

III. Measurement of soil temperature

• Make a hole in the soil with an augar to the depth at which temperature is to be
measured.

• Place on junction at that depth and re-pack the soil to ensure proper contact.

• Dip the other end in the melting ice.

• Measure the emf with micro voltmeter (potentiometer).

• Calculate the temperature from the calibration curve. Then no couples can be
joined in series of parallel. In series they are called thermopiles and output will be
sum of emf of all the thermocouples. Mostly these are joined in series as they
increase the sensitivity of the system.

78
Observations and calculations

Potentiometer reading = X micro volt

Slope of the Calibration curve = 40 microvolt ° C-1

Temperature = X
------- °C
40

Result:

SI. No. Soil depth cm emf recorded Microvolt Temperature °C

79
IV. Soil Temperature using Soil thermometer

Soil temperature has a significant role in helping to determine the rate of plant
growth, and whether a plant will even survive. The temperature in the soil changes
greatly with depth. To measure soil temperature, find an area that is not in direct sunlight.
Using a thermometer, measure the air temperature at shoulder height. Hold the
thermometer still for about one minute (make sure your fingers are not on the
thermometer bulb), read and record the air temperature. Next, measure the temperature at
the surface of the ground. Put the thermometer flat on the ground and record the
temperature after one minute. To determine the temperature below the ground surface,
use a dowel that you have marked at 1 inch, 2 inches, 6 inches and 12 inches. Start by
pushing the dowel into the ground till you reach the 1 inch mark. Remove the dowel and
insert the thermometer for one minute, then remove the thermometer and quickly record
the temperature. Repeat this procedure to obtain temperature readings at 2 inches, 6
inches and 12 inches. Take temperature readings at different times throughout the day at
the same location. To compare with soil temperatures for areas in direct sun, just repeat
using the same procedure but select an area that gets full sun. You will note that the soil
temperatures in these areas are typically much higher than in the shaded areas.

Soil Temperature Conditions during growing season

Less than 40° F No growth, bacteria and fungi are not very active

40° F to 65° F Some growth

65° F to 70° F Fastest growth

70° F to 85° F Some growth

above 85° F No growth

80
 2.11 Estimation of Moisture Content in Soil

Aim: To determine the moisture content of the given samples

Principle

Moisture content is determined by heating a known weight of the sample in an

electric oven at 105°C until it attains a constant weight. The loss in weight is reported to
be the moisture content of the sample.

Apparatus required

1. Moisture bottle

2. Balance

3. Desiccator

Procedure

Place a weighing bottle with its stopper separately in an electric oven at 105°C for
about 15 minutes. Replace the stopper, remove the bottle, cool in a desiccator and record
the weight. Transfer about 50 g of sample into the bottle, fix the stopper and weigh
quickly. Remove the stopper and keep it in the oven at 105°C for 8 hours. After the
expiry of time replace the stopper, remove the bottle from the oven, cool it in a desiccator
and weigh quickly. Calculate the loss in weight and express it in percentage.

Calculation

Weight of moisture bottle = Ag

Weight of moisture bottle+ sample = Bg
Weight of sample alone = (B-A) g
Weight of moisture bottle+ sample) = Cg
after drying in the oven
Weight of moisture in the sample = (B-C) g
Percentage of moisture in the sample = (B-C)
-------- × 100
(B-A)
Result:

81
 2.12 Determination of Soil Texture

(International Pipette Method/ Robinson's Pipette Method)

Aim: To determine soil texture by International Pipette Method/ Robinson's Pipette

Method in the given soil sample

Principle

This method is based on Stokes' law. According to this law the rate of fall of a
particle in liquid is directly proportional to the square of its radius. V α r2

V = 2 / 9 gr2(dp-d)/ŋ

Where

V = Sedimentation velocity in cm/sec

g = Acceleration due to gravity cm/sec

r = Radius of the particle or sphere (cm)

dp = Density of the particle (g/cc)

d = Density of the liquid (g/cc)

n = Viscosity of the liquid

The soil is first dispersed by destroying the binding agents with hydrogen
peroxide and hydrochloric acid followed by treatment with a dispersing agent. Clay and
silt are separated by sedimentation and coarse and fine sand by sieving.

Apparatus required

1. 1000 mL spoutless measuring cylinder

2. Rubber stopper for the cylinder

3. 500 mL beaker

4. Mechanical stirrer

5. Funnel (7.5cm)

82
5. Filter paper Whatman No.50

6. Filter stand

7, 150 mL measuring cylinder

8. Water bath

9. Stop clock

10. Chemical balance

11. Tall form beaker

12. Robinson pipette

13. Hot air oven

14. Porcelain dish/basin

Reagents required

1. 6% Hydrogen peroxide

2. 0.2N Hydrochloric acid

3. 1N Sodium hydroxide

Procedure

Transfer exactly 20 g of air dried soil sample to a 500 mL beaker. Add 60 mL of

6 per cent hydrogen peroxide. Stir it well and keep it on a water bath for 30 minutes till
frothing ceases. Treatment with hydrogen peroxide is to destroy the organic matter which
is binding soil particles. Hydrogen peroxide treatment is not necessary when the organic
matter content of the soil is negligible. Then add 200 mL of 0.2 N HC1, stir it well and keep
it over night. Hydrochloric acid is added to destroy CaCO3 which is also binding agent.

Filter the contents through, what man No.50 filter paper and wash it with water
till the filtrate runs free of chloride (collect about 10 mL of the filtrate in a test tube and
add 2-3 drops of silver nitrate solution. Formation of curdy white precipitate indicates the
presence of chloride). When the filtrate runs free of chloride, transfer the soil material
from the filter paper to another 500 mL beaker and add about 400 mL water. Then add
8 mL of normal sodium hydroxide and stir it well for 10 minutes with a mechanical
83
stirrer. Transfer the contents to a 1000 mL spout less measuring cylinder and make up to
1000 mL mark with water. Cover the cylinder tightly with a rubber stopper and shake the
contents thoroughly by repeated inversions holding the rubber stopper tightly so as to
avoid spilling of the soil water suspension.

Clay and silt

Remove the rubber stopper and place the cylinder under Robinson pipette and
start a stop clock simultaneously. Note down the temperature and the settling time for
clay plus silt from the table. Till the settling time is over do not disturb the suspension.
First lower the pipette in such a way that the tip of the pipette just touches the surface of
the suspension. At the end of the stipulated settling time for clay and silt, lower the
pipette to 10 cm depth and draw 20 mL suspension and deliver it to a weighed clean
porcelain dish. This suspension contains clay plus silt. Evaporate this first by keeping in
on a water bath and dry it in an air oven at 105°C. Cool it in a desiccator and determine
the weight of clay plus silt and calculate the percent.

Clay alone

Shake the contents of the cylinder well and allow undisturbed till the stipulated
settling time for clay alone corresponding to the suspension temperature. Withdraw 20
mL of the suspension at the end of the period as done in the case of clay plus silt and
determine the weight as clay alone after evaporating and drying.

Coarse sand and Fine sand

Pour out major portion of the suspension from the cylinder after with drawing
sample for clay alone. While pouring out care should be taken to see that no sand fraction
is lost. Then wash the sediment with water and transfer the contents to a tall form beaker.
Add water to a height of more than 10 cm. Stir well and allow it to stand for 4 minutes.
Then pour off the supernatant liquid. Repeat this process till the water poured off, is no
longer turbid. Transfer the residue to a porcelain basin, dry it in an oven and weigh as
coarse sand plus fine sand. Sieve the coarse sand and fine sand in a seventy mesh sieve.
The material passing through the sieve will be fine sand while the coarse sand fraction
will be retained on the sieve. Weigh it as coarse sand

84
Calculation

Clay + silt

Weight of Soil taken = 20 g

Volume of Suspension = 1000 mL

Volume of Suspension pipetted out = 20 mL

Weight of empty porcelain dish = ag

Weight of silt + clay + dish + NaOH = bg

Weight of clay + silt + NaOH = (b-a) g

Weight of NaOH alone = 0.0064 g

(Present in 20 mL of suspension)

Weight of clay + silt alone = b – (a+0.0064) g

Per cent clay + silt = b – (a+0.0064) ×1000 × 100

------------------------------------
20 × 20

Clay alone

Weight of empty porcelain dish = qg

Weight of dish + clay + NaOH = pg

Weight of clay- NaOH = (q-p) g

Weight of clay alone = q - (p + 0.0064) g

Weight of NaOH alone (in = 0.0064 g

20 mL of suspension)

Per cent clay = q - (p + 0.0064) × 1000 × 100

--------------------------------------
20 × 20

Per cent silt = (Per cent clay + silt) - (Percent clay)

85
Coarse sand + fine sand

Weight of porcelain basin = xg

Weight of dish + coarse sand + fine sand = yg

Weight of coarse sand + fine sand alone = y-xg

Per cent coarse sand + fine sand = (y – x) × 100

------------------
20

Coarse sand alone

Weight of porcelain basin = cg

Weight of dish + coarse sand = dg

Weight of coarse sand alone = c-dg

Per cent coarse sand + fine sand = (c – d) × 100

-----------------
20

Per cent fine sand = (Per cent coarse sand + fine sand) – (Per cent coarse sand)

Sedimentation time for clay and silt fractions (Depth of sedimentation = 10 cm)

Sedimentation Time for clay Sedimentation Time for silt

Temperature °C
Hrs Min Min Sec

8 11 ---- 6 40

9 10 40 6 30

10 10 25 6 20

11 10 10 6 10

12 9 60 6 01

13 9 35 5 50

14 9 20 5 40

86
 Sedimentation Time for clay Sedimentation Time for silt
Temperature °C
Hrs Min Min Sec

15 9 05 5 30

16 8 50 5 20

17 8 35 5 10

18 8 25 5 05

19 8 10 5 0

20 8 0 4 48

21 7 50 4 40

22 7 40 4 30

23 7 25 4 30

24 7 15 4 - 20

25 7 05 4 15

26 6 55 4 10

27 6 45 4 05

28 6 40 4 0

3 6 30 3 55

30 6 20 3 50

31 6 05 3 40

32 5 55 3 35

87
 3. SOIL CHEMICAL ANALYSIS

3.1 Determination of Soil pH and Soil Electrical Conductivity (Jackson, 1973)

Aim

To determine the pH and electrical conductivity of the given soil sample.

Soil pH

pH is defined as the negative logarithm of hydrogen ion activity at 25ºC (or) the
log of the reciprocal of the hydrogen ion concentration (Sorenson, 1906).

pH = -log(H+) = log 1/(H+)

Two electrodes are used in the determination of pH. One is reference electrode
which provides standard voltage. The reference electrode is usually a saturated calomel
electrode which has two layers (i) saturated solution of KCl and (ii) mixture of solid
HgCl2 and Hg. The outer tube is usually 5-15 cm long, 0.5-1 cm in diameter. The mixture
of solid HgCl2 + Hg paste is contained in an inner tube that is connected to the saturated
KCl solution in the outer tube by means of a small opening. The resistance of this type of
electrode is 2000-3000 ohms.

The outer electrode is a glass electrode that consists of a tube enclosing a lead
wire made of Ag coated with AgCl. This wire is again enclosed in a wax insulation.
To the tube at the bottom is attached a glass bulb made of a special kind of glass which is
sensitive to H ions. The thickness of the glass membrane varies from 0.03 to 0.1 mm and
has a resistance of 50 to 500 mega ohms.

When these two electrodes are dipped in solution, the saturated solution of KCl
comes out of the reference electrode though the small holes and forms an invisible ionic
bridge between electrodes through which current passes. The H ions are absorbed by
glass electrode and depending on the amount of H ions present in the solution, an electric
potential develops between the electrodes. The potential difference is measured in terms
of pH by suitable galvanometer.

88
Principle

Glass electrode on contact with H ions of the solution acquires an electrode potential
which depends on the concentration of H ions. This is measured potentiometrically against
some reference electrode which is usually a calomel electrode. The potential difference
between glass electrode and calomel electrode is expressed in pH units.

Materials required

i. pH meter

ii. 100 ml beakers

iii. Glass rods

iv. Buffer solution (pH 4.0, 7.0 and 9.2)

Procedure

x Switch on the instrument and allow it to warm for 10 minutes

x Keep the pH selector switch on zero position

x Set the temperature compensation control to the solution temperature

x Adjust the zero adjustment knob so that the pointer in the meter reads exactly zero,
when the electrodes are immersed in distilled water

x Lift the electrodes from distilled water and wipe with filter paper and dip them in
standard buffer solution of known pH (4.0, 7.0 and 9.2)

x Change the function switch to a particular pH range (0.7 or 7-14) and adjust the
standardization knob till the pointer reads the correct pH value of the buffer solution.
Do not disturb the zero knob adjustment

pH measurement

x Weigh 20 g of air dry soil passed through 2 mm sieve and transfer to a clean 100 ml
beaker

x Add 50 ml of distilled water

x Using glass rod, stir the contents intermittently and allow it to stand for half an hour

89
x Wash the electrodes carefully with a jet of distilled water and wipe with a piece of
filter paper

x Stir the soil suspension again just before taking the reading

x Immerse the electrodes into the beaker containing soil water suspension and change
the function switch the particular pH range

x Record the meter reading both in supernatant solution and suspension

Precautions

x Electrodes should not be rubbed strongly with hand or any other material as they are
too fragile

x Electrodes should not be allowed to dry. They should be kept in distilled water ehen
not in use

x Electrodes should not be allowed to remain in the test solution for a long time. After
testing it should be washed with distilled water immediately.

Rating

<4.5 : Extremely acidic

4.5-5.0 : Very strongly acidic

5.1-5.5 : Strongly acidic

5.6-6.0 : Moderately acidic

6.1-6.5 : Slightly acidic

6.6-7.3 : Neutral

7.4-7.8 : Mildly alkaline

7.9-8.4 : Moderately alkaline

8.5-9.0 : Strongly alkaline

>9.0 : Very Strongly alkaline

90
 SOIL ELECTRICAL CONDUCTIVITY

Electrical conductivity (EC) gives the total amount of soluble salts present in the
soil and is expressed as dSm-1

It is known that solutions offer some resistance to the passage of electric current
through them, depending upon the concentration of salts present. Hence EC is measured
in terms of electrical resistance between parallel electrodes immersed in the soil
suspension of water. In such a system, the solution between the electrodes becomes the
electrical conductor to which the physical laws relating the resistance are applicable. The
electrical resistance “R” is directly proportional to the distance “L” between the
electrodes and inversely proportional to the cross sectional area “A” of the conductor.

Hence R vL/A (or) R = r x L/A

Where r = proportionality constant known as electrical resistivity

If L = 1 cm and A =1 cm2 then R =r where r is called specific resistivity

Hence specific resistance is the resistance of conductor of 1 cm in length and

2
1 cm in area. Higher the salt content, higher the passage of current and lesser is the
resistance to the flow of the current. Hence the reciprocal of specific resistivity is called
specific conductivity.

Specific conductivity is defined as the conductivity of a solution enclosed in a cell

whose electrodes are exactly 1 cm and posses a surface area of 1 cm2. The resistance is
expressed as ohms /cm and the conductivity is expressed in reciprocal ohms or mhos / cm.
It is not possible to make a conductivity bridge having electrodes of 1 cm2 area placed
exactly 1 cm apart. Hence the factor called cell constant is determined for the given cell.

Principle

As the amount of soluble salts in a solution increases the electrical conductivity

also increases. The electrical conductivity is measured in terms of the resistance offered
to the flow of current using a conductivity bridge.

91
Procedure

x Switch on the electrical conductivity bridge and wait for 10 minutes

x Check the instrument with saturated CaSO4 and 0.01 N KCl solutions. EC should be
2.2 and 1.41 dSm-1 respectively.

x Use the same soil water suspension used for measuring pH for the estimation of EC.

x Stir the content and allow the soil to settle for 10 minutes.

x Wash the electrodes carefully and immerse them into soil solution

x Adjust the temperature condition. Adjust the meter knob until the magic eye of the null
indicator is at the widest width. The readings on the scale at this position indicate the
electrical conductivity. Multiply this by the cell constant (noted on the cell itself) to
get specific conductivity.

92
 3.2 Estimation of available Nitrogen in soil by Alkaline Permanganate method

(Subbiah and Asija, 1956)

Aim

To estimate the quantity of available (mineralisable) nitrogen in the given soil

sample.

Principle

The available nitrogen in the soil is estimated by distilling the soil with 0.32%
KMnO4 and 2.5% NaOH. The evolved ammonia is absorbed in 2% boric acid and titrated
against standard sulphuric acid. The amount of nitrogen that is mineralized in this
estimation is considered as available for crop uptake.

Reactions

2KMnO4 3O + K2O +2 MnO2

Organic matter Release of R-NH2

R-NH2 + 3 O R-OH - NH2

Reagents

1. 0.32% KMnO4 solution

2. 2.5% sodium hydroxide

3. Liquid paraffin

4. 0.02 N H2SO4

5. 2% Boric acid solution

6. Double indicator (prepared by dissolving 0.066 g of methyl red and 0.099 g of

bromocresol green in 100 ml of ethyl alcohol)

Procedure

x Transfer 20 g of 2 mm sieved soil into 1 litre distillation flask.

x Add 20 ml of water, 1 ml of liquid paraffin and a few glass beads.

93
x Add 100 ml each of freshly prepared 0.32% KMnO4 solution and 2.5% sodium
hydroxide solution.

x Distill the contents and collect about 100 ml distillate in a 250 ml beaker containing
20 ml of 2% boric acid solution with 2 drops of double indicator.

x Calculate the amount of mineralisable (available) nitrogen present in the soil.

Calculations

Weight of soil taken : 20 gm

Volume of 0.02 N H2SO4 used : A mL

1 ml of 0.02 N H2SO4 : 0.00028 g of N

x ml of 0.02 N H2SO4 : 0.00028 x A g of N

This is present in 20 g of soil

In 2 x 106 kg soil i.e., the weight of 1 hec

furrow slice soil

0.00028 x A x 2.24x 106

= ---------------------------------- kg ha-1
20
Rating of soil with reference to available N status

Low = Less than 280 kg ha-1

Medium = 280-450 kg ha-1

High = More than 450 kg ha-1

94
 3.3 Estimation of Available Phosphorus in Soil by Olsen and Bray No.1 Method
(Olsen et al., 1954)

Aim

To estimate the available phosphorus status in the soil by Olsen and Bray No.1
methods.

Principle

Olsen method

This method of extraction of available soil phosphorus is suited for calcareous and
alkaline soils. The CO32 – ions from NaHCO3 will react with Ca 2+ and CaCO3is precipitated,
thus allowing the same into the solution. The amount of P extracted is determined
colorimetrically.

Bray-1 method

This method of extraction of available soil phosphorus is suited for acid and neutral
soils. The combination of HCl and NH4 F, extracts acid soluble forms of P. The fluoride ion
complexes Al3+ and Fe3+, thus allowing P to come into the solution. The amount of
P extracted is determined colorimetrically.

Reagents

Olsen method

1. Olsen reagent – 0.5 M NaHCO3 at pH 8.5

2. Darco G-60

3. Ammonium-molybdate

4. Sulphuric acid

5. Ascorbic acid

Bray No.1 method

1. Bray No.1 extractant – 0.03 M NH4F in 0.025 N HCl

2. Ammoniummolybdate

95
 3. Sulphuric acid

4. Ascorbic acid

Procedure

Olsen method

Take 5 g of the 2 mm sieved soil and transfer it to a 250 ml shaking bottle. Add
50 ml of the Olsen’s reagent and a pinch of Darco G 60 (To make the solution
colourless). Shake in a horizontal shaker for 30 minutes. Filter the contents through
Whatman No.1 filter paper collecting the filtrate in a suitable container. Estimate the P
concentration in this extract by the ascorbic acid method.

Bray No.1 method

Take 1 g of the 2 mm soil and transfer in to a 250 ml shaking bottle. Add 50 ml of

the Bray No.1 extractant and shake in a horizontal shaker for 1 minute. Filter the contents
through Whatman No.1 filter paper collecting the filtrate in a suitable container.
Estimate the P content in the extract by ascorbic acid method.

Colorimetric estimation of P by ascorbic acid method (Murphy and Riley, 1962)

Reagents

1. Ammonium molybdate-Antimony potassium tartarate solution (Stock solution)

Dissolve 12 g of ammonium molybdate in 200 ml of warm water and cool.

Dissolve 0.291 g of antimony potassium tartarate in100 ml distilled water. Add both the
solutions to 1000 ml of 5 N H2SO4 (138 ml of con. H2SO4 per litre of water), mix
homogeneously and make-up to 2 litres with distilled water. Store in a pyrex glass bottle
in a dark and cool compartment. This solution is heat and light sensitive (Reagent A).

2. Ascorbic acid reagent

Dissolve 1.056 g of ascorbic acid in 200 ml of reagent A. This reagent should be

prepared freshly as and when required (Reagent B).

96
Procedure:

Pipette out 5 ml aliquot of the soil extract of either Olsen or Bray method into a
25 ml volumetric flask. Adjust the pH to 5 by taking 5 ml of the Olsen or Bray No.1 soil
extract and assessing the volume 1 N H2SO4 or 1 N NaOH required to adjust the pH to 5.
Aqueous 0.25% solution of p-nitrophenol can be used as indicator which will be yellow
at pH 5 and above and colourless at pH below 5.

After having adjusted the pH to 5, add 4 ml of the ascorbic acid reagent (reagent
B), makeup the volume, mix well and allow for 20 minutes. Measure the intensity of
blue colour in a photoelectric colorimeter at 660 nm (red filter). Prepare a standard curve
with standard P solutions. Find out the concentration of P in the soil sample extract using
the standard curve and calculate the available P status in the soil from the P concentration
in the soil extract.

Preparation of standard curve

Standard P solution

Prepare 100 ppm P solution by dissolving 0.4281 g of AR grade KH2PO4 in

distilled water and making up the volume to 1000 ml. Dilute 10 ml of the 100 ppm P
solution to 100 ml to prepare 10 ppm P solution.

Pipette out required volumes of the standard P solutions into 25 ml volumetric

flasks as given below and add 5 ml of the respective reagent (Olsen or Bray No.1
reagent). Adjust the pH to 5 using paranitrophenol indicator and 1 N H2SO4 or 1 N
NaOH as the case may be. Add 4 ml of the ascorbic acid reagent (reagent B), make up the
volume to the mark and allow 20 minutes for colour development. Measure the intensity
of the blue colour in a photoelectric colorimeter at 660 mm (red filter) and draw the
standard curve with the P concentrations in the X axis and the % T or O.D. in the Y axis.

97
 Volume to be
P in ppm Volume of standard solution
made up

0.2 0.5 ml 10 ppm solution 25 ml

0.4 1.0 ml 10 ppm solution 25 ml

0.6 1.5 ml 10 ppm solution 25 ml

0.8 2.0 ml 10 ppm solution 25 ml

1.0 2.5 ml 10 ppm solution 25 ml

1.2 3.0 ml of 10 ppm solution 25 ml

1.4 3.5 ml of 10 ppm solution 25 ml

1.6 4.0 ml of 10 ppm solution 25 ml

Calculations

Weight of soil used for extraction = 2g

Volume of extractant used = 50 ml

Volume of the extracted solution taken for P estimation = 5 ml

Final volume was made upto = 25 ml

Concentration of P read in the standard curve = a ppm

corresponding to the present transmittance

Therefore, amount of available P in 1 hectare of the soil

a 25 2.24 x 106
= ------- x --- x 50 x ----------- kg ha-1
106 5 5
Rating of soil with reference to available P status

Low = Less than 11 kg ha-1

Medium = 11 - 22 kg ha-1

High = More than 22 kg ha-1

98
 3.4 Estimation of available Potassium in Soil

(Stanford and English, 1949)

Aim

To estimate the available potassium status in the given soil sample.

Principle

The method is based on the principle of equilibrium of soils with an exchanging

cation made of the solution of neutral normal NH4OAc, in a given soil solution ratio.
During the equilibrium, ammonium ions exchange with the exchangeable K ions of the
soil. The K content in the equilibrium solution is estimated with a flame photometer.
Since NH4+ holds highly charged layers together just as K, the release of the fixed K, in
an exchangeable form, is retarded during NH4OAc extraction

K+ - Clay + NH4- NH4- - Clay + K+ + excess of NH4-

Reagents

i. Neutral 1 N NH4OAc solution

ii. Standard K solution

Apparatus required

1. 100 mL polythene shaking bottle

2. 25 mL measuring cylinder

3. Reciprocating shaker

4. Whatman No.40 filter paper

Procedure

Take 5 g of the soil sample in a 250 ml shaking bottle. Add 25 ml of the neutral N
NH4OAc extractant and shakefor 5 minutes in a horizontal shaker. Filter the contents
through Whatman No.1 filter paper collecting the filtrate in a suitable container. Estimate
the K concentration in the extract by aspirating the solution into the flame of the flame
photometer and measuring the reading. Prepare a standard curve for K concentrations

99
ranging from 0 to 100 ppm at 10 ppm interval. Find out the K concentration of the soil
extract from the standard curve and calculate the available K status in the soil.

Calculations

Weight of soil taken = 5g

Volume of 1 N NH4OAc extractant added = 25 ml

K concentration in the soil extract = X ppm

Available K status in the given soil

X x 25 2.24 x 106
--------- x ----------
106 5
Rating of soil with reference to available K status

Low = Less than 118 kg ha-1

Medium = More than 118 and upto 280 kg ha-1

High = More than 280 kg ha-1

100
 3.5 Estimation of available Sulphur in soil

(Williams & Steinbergs, 1959)

Aim

To estimate the available sulphur content in the given soil sample

Principle

Sulphate in the soil is extracted with calcium chloride and converted into barium
sulphate by adding solid barium chloride. Sulphur is estimated turbidimetrically in a
colorimeter using the blue filter or in a spectrophotometer at 340 nm.

Apparatus required

1. 100 ml shaking bottle

2. Reciprocating shaker

3. Measuring cylinders - 25 ml and 10 ml

4. 25 ml volumetric flask

5. Pipettes of 5 ml and 1 ml capacity

6. Spectrophotometer / photoelectric colorimeter

Reagents required

1. 0.15 % Calcium Chloride dihydrate (CaCl2.2H2O)

Dissolve 1.5 g of CaCl2.2H2O in about 700 ml of distilled water and make to 1

litre with distilled water

2. Barium chloride dihydrate (BaCl2.2H2O)

Barium chloride crystals passed through 20 to 30 mesh sieve

3. Conditioning agent

Dissolve 75 g of NaCl in 275 ml distilled water in a 500 ml volumetric flask,

stirring with a magnetic stirring bar and add 30 ml of conc. HCl, 100 ml of
absolute alcohol and 50 ml glycerol into the flask. Continue stirring until NaCl
dissolves. Remove stirring bar and make the volume with distilled water.

101
 4. Standard sulphate solution

Dissolve 0.5434 g of K2SO4 in 1 litre of distilled water. This contains 100 ppm of
S. Take 10 ml of 100 ppm solution and dilute 10 times to get 10 ppm solution of
S. “From the above standard, various working standards are prepared as
mentioned below. Run a blank simultaneously. Add 2.5 ml of conditioning agent
and 0.5 g of BaCl2 crystals and measure the turbidity at 340 nm.

S in ppm Volume of standard solution Volume to be made up

1.0 3 ml of 10 ppm solution 30 ml

2.0 6 ml of 10 ppm solution 30 ml

3.0 9 ml of 10 ppm solution 30 ml

4.0 12 ml of 10 ppm solution 30 ml

5.0 15 ml of 10 ppm solution 30 ml

6.0 18 ml of 10 ppm solution 30 ml

7.0 21 ml of 10 ppm solution 30 ml

8.0 24 ml of 10 ppm solution 30 ml

9.0 27 ml of 10 ppm solution 30 ml

10.0 30 ml of 10 ppm solution 30 ml

Procedure

¾ Weigh five grams of soil into a 100 ml shaking bottle

¾ Add 25 ml of 0.15% CaCl2 and shake for 30 minutes in a reciprocating shaker.

¾ Filter the suspension through Whatman No. 42 filter paper

¾ Pipette out 10 ml extract into a 50 ml volumetric flask and add 20 ml of water thus
bringing the volume to 30 ml

¾ Add 2.5 ml of conditioning agent and 0.2 to 0.5 g of BaCl2 crystals by a spatula to
the extract

102
¾ Shake the contents of the flask for 1 minute each at a constant rate

¾ After 1 to 3 mts, measure the turbidity in a colorimeter using a blue filter

¾ Turbidity can be more accurately measured at 340 nm on a spectrophotometer

¾ Turbidity remains constant during 3-10 mts..

Calculations

Weight of soil sample = 5g

Volume of 0.15 % CaCl2 added = 25 mL

Volume of extract taken for analysis = 10 mL

Volume made up = 30 mL

Concentration of ‘S’ read from the standard = A ppm

curve corresponding to the per cent absorbance

SO4-S (mg kg-1) = A X 25 X 2.24 X 106 ppm

106 X 5 X 10

103
 3.6 Estimation of Available Micronutrients

(Lindsay and Norvell, 1978)

Several extractants have been tried to extract and estimate the available
micronutrient status of the soil. Of this Diethylene Triamine Penta Acetic Acid (DTPA)
was found to be the most suitable extractant for estimating the content of available Zn,
Cu, Mn and Fe by the use of atomic absorption spectrophotometer.

Principle

DTPA forms stable complexes with Zn, Cu, Mn and Fe. Its capacity to complex
each of the micronutrient cation is 10 times its atomic weight.

Materials required

(i). DTPA 0.005 M, (ii) CaCl2.2H2O – 0.01 M solution and (iii) Triethanolamine: 0.1 M
solution. (TEA suppresses the solubility of CaCO3 which will release the occluded
micronutrients unavailable to the plant).

DTPA extractant solution is prepared by dissolving 13.1 ml of TEA, 1.967 g of

AR grade DTPA and 1.47 g of CaCl2 in 100 ml glass distilled water. The contents are
allowed for some time so that the DTPA will dissolve and then dilute to about 900 ml.
The pH of the solution is adjusted to 7.3 ± 0.05 with 1:1 HCl by stirring and the volume
made upto 1 litre.

Preparation of standard solution

Zinc

0.4415 g of AR grade ZnSO4.7 H2O is dissolved in 200 ml glass distilled water in a

beaker to which 5 ml of 1:5 H2SO4 is added. The contents are transferred to a one litre
volumetric flask and the volume made up. This gives 100 ppm of Zn. From this stock solution
working standards of 0, 0.1, 0.2, 0.4 and 0.6 μg/ml (or) ppm solutions are prepared.

Iron

0.702 g of AR grade ferrous ammonium sulphate dissolved in 5 ml of 1:5 H2SO4 and

made up to 1 litre with distilled water gives 100 ppm of Fe stock solution. From this stock
solution various concentrations of working standards viz., 0, 1, 2, 4, 6 ppm are prepared.

104
Manganese

0.288 g of AR grade potassium permanganate is dissolved in 300 ml of glass

distilled water. To this 20 ml of conc.H2SO4 is added and warmed to 60oC. Then oxalic
acid is added drop by drop to make the solution colourless. The contents are cooled and
made upto 1 litre with gives 100 ppm stock solution. From these working standards of 0,
1, 2, 4, 6, 8 ppm are prepared.

Copper

0.392 g of AR grade CuSO4 5H2O dissolved in glass distilled water and made
upto 1 litre gives 100 ppm of Cu. From this working standards of 0,1,4,6 ppm solutions
are prepared.

From the working standard solutions, standard curve for each nutrient is prepared.
While preparing the standard, start from the lowest concentration just after standardizing
the instrument with blank solution. In between two standard solutions, introduce the
blank and ensure that there is no change in the zero point. Prepare a graph by plotting the
absorbance values against concentrations.

Procedure

x Transfer 10 g of air dried soil sample to 150 ml conical flask / polythene bottle and
add 20 ml of the DTPA extractant solution.

x Close the bottle and shake for 2 hours in a horizontal shaker.

x Filter through Whatman No.42 filter paper.

x Take the reading of the filtrate in the atomic absorption spectrophotometer and by
referring to the standard curve calculate the concentration of each micronutrient in
the sample.

x Shaking time, concentration and pH of the DTPA extractant and temperature will
influence the quantity of nutrients extracted.

105
Critical levels of micronutrients

ƒ Zn 1.2 ppm,

ƒ Cu 0.72 ppm

ƒ Fe 3.7 ppm (Non Calcareous soil)

6.3 ppm (calcareous soil)

ƒ Mn 2.0 ppm.

106
 4. FERTILIZER ANALYSIS

4.1 Fertilizer Sampling Technique

Requirements of sampling

The following measures and precautions should be observed while drawing

fertilizer samples.

x Samples should not be taken during adverse weather conditions.

x The sampling instruments should be clean and dry.

x Contaminations should be avoided while sampling.

x Representative samples should be drawn by thoroughly mixing the contents of

each fertilizer container selected for sampling.

x The samples should be stored in a suitable clean, dry and air tight containers.

x The sampling container should be completely filled with sample.

x The sampling container should be sealed air tight with proper labeling,
carrying particulars such as dates of sampling etc.

x Samples should be stored in a cool place.

Sampling from packages or containers

x All containers in a single consignment of material of the same grade and type
from a single batch constitute a lot.

x If a consignment is containing different batches, the containers of each batch

shall constitute a separate lot.

x If a consignment is drawn in a continuous process, 2000 containers or

100 tonnes of the material shall constitute a lot.

107
Number of containers to be selected from a lot

No. of containers to be
Lot size (N)
selected (n)
Less than 10 1
10-100 2
100-200 3
200-400 4
400-600 5
600-800 6
800-1000 7
1000-1300 8
1300-1600 9
1600-2000 10

Random number tables may be used while selecting the containers at random.
In case, the random number table is not available, count the container as 1, 2, 3 upto r and
so on, r being equal to the integral part of N/n. Every rth container shall be withdrawn
which will constitute the sample.

Composite samples

x Draw portions of the material from different parts of each container using appropriate
sampling instrument. Mix thoroughly and make into a composite sample.

x If the material is packed in containers which do not permit sampling instruments,

empty the contents on a clean hard surface and draw composite sample by quartering.

x A composite sample of 2 kg weight is always desirable. If the gross sample is more

than 2 kg, reduce it to 2 kg by quartering. Spread the composite sample over a clean
hard surface and divide into four equal parts. Reject diagonally opposite parts and
mix the remaining two parts together to form a cone, flatten out the cone and repeat
the quartering till a composite sample of 2 kg weight is obtained.

108
Sampling from a bulk in heaps or wagons

Scale of sampling: Take samples from each heap or wagon as case may be.

Composite sample: Draw samples from each heap or wagon by means of a scoop from
different parts viz., the front, middle and back and at different depths. Reduce the bulk to
a desired level by quartering.

Test and reference samples

Prepare test sample of about 0.5 kg from the composite sample. Spread the
composite sample on a clean hard surface and divide into three equal portions of not less
than 0.5 kg each. Each of these samples constitutes the sample.

Transfer each test sample into a suitable container and close with tight fitting
stopper or lid so that the original composition remains unchanged. Seal the containers
with seals of inspecting officer and the manufacturer or dealer.

Send one sample to State Fertilizer Analyzing Laboratory and the second to
manufacturer/dealer. The third sample is retained by the Inspecting Officer for production
in the court, if necessary.

Preparation of samples for lab analysis

x Reduce the test sample to a quantity sufficient (250 g) for analysis.

x Sieve through 1 mm or No. 20 standard sieve for fertilizer materials that form a
paste on putting pressure while grinding with porcelain pestle and mortar.

x Use No. 40 standard sieve for dry mixtures that tend to segregate while grinding in
a porcelain pestle and a mortar.

x Grind as rapidly as possible to avoid loss or gain of moisture during operation.

x Mix thoroughly and store in a tightly stoppered bottle.

(The Fertilizer Control Order, 1985 as amended upto 1998.)

109
 4.2 Estimation of Nitrogen in Urea

Aim

To estimate nitrogen content in the given fertilizer sample - urea

Principle

The amide form of N present in urea is converted to ammoniacal nitrogen

(ammonium sulphate) when it is digested with concentrated sulphuric acid. The
ammoniacal N in the digested material is liberated when it is distilled with a strong alkali.
The liberated ammonia is absorbed in a known excess standard acid and the actual volume of
standard acid used is determined by back titrating against standard alkali. From the actual
volume of standard acid consumed, the N content of the urea is determined.

Reaction:

CO (NH2)2 + H2SO4 (NH4)2SO4 + SO4 + CO2 + H2O

Materials required

(i) Concentrated sulphuric acid (ii) N/10 Sulphuric acid (iii) N/10 KOH (iv) Potassium
sulphate (v) Copper sulphate (vi) 40% NaOH (vii) 10% Sodium sulphide (viii) Zinc
granules and porcelain bits (ix) Kjeldahl flask (x) Volumetric flask 500 ml and
(xi) Funnel.

Procedure

x Weigh exactly 1 g of the fertilizer sample and transfer into a Kjeldahl digesting flask
taking care to see that no fertilizer granule sticks to the neck of the flask.

x Add about 30ml of concentrated sulphuric acid mix well.

x Add the digestion mixture – about 2g of potassium sulphate and about 0.2g of copper
sulphate(10:1 ratio)

[Addition of potassium sulphate increases the boiling point of the acid and thereby
the digestion time is reduced. Copper sulphate acts as a catalyst to hasten the reaction
and also it serves as an indicator (appearance of apple green colour indicates the
completion of digestion)]

110
x Digest the material till it turns into apple green.

x Transfer the whole digested material,after cooling into a 500 ml volumetric flask with
the help of a funnel without spilling.Rinse the Kjeldahl flask with about 50ml of
water 3 or 4 times and transfer this into the volumetric flask. Then make up the
volume and shake the contents thoroughly so as to get a homogeneous solution.

x Pipette out 50ml of this solution into a distillation flask and add about 500- 600ml of
water. Add few zinc and porcelain bits (Addition of zinc keeps the contents of the
flask in a reduced condition and also maintains the pressure at a higher level).

x Add about 20 – 30 ml of 40% NaOH till the contents become distinctly alkaline as
tested with a litmus paper. Then add about 10ml of sodium sulphate (Copper or
mercury will adsorb, if present, the ammonia liberated during distillation. Copper
forms cupra ammonium complex. The ammonia bound in this way will not be
released during distillation.When sodium sulphate is added, the copper is converted
to its sulphate. Sulphides of copper will not bind ammonia and hence the addition of
sodium sulphide along with 40% NaOH).

x Start distillation after placing a known excess of N/10 sulphuric acid at the delivery
end and determine the actual volume of N/10 sulphuric acid consumed and calculate
the percentage N.

Note: Do not remove or lower the flame during distillation. This leads to back
suction of the standard acid kept at the delivery end.

Calculations

Weight of fertilizer sample taken = 1g

Volume made up after digestion = 500 mL

Volume of aliquot pipetted out for distillation = 50 mL

Volume of N/10 H2SO4 taken in the ice tumbler = 25 mL

Volume of N/10 KOH used for back titration = X mL

111
Actual volume of N/10 H2SO4 consumed = (25-X) mL

1 ml of N/10 H2SO4 = 0.0014 g N

Therefore (25- X) mL of N/10 H2SO4 = 0.0014x (25-X)g N

This is from 50 mL of aliquot = 0.0014 x (25- X) x 500

-----
50

This is present in 1 g of fertilizer sample

Therefore percentage of total N in the given = 0.0014 x (25- X) x 500 100

sample
----- × -----
50 1

112
 4.3 Estimation of NH4 – N and NO3 – N in Ammonium Nitrate

Aim

To estimate the contents of ammoniacal and nitrate forms of N separately in

ammonium nitrate / calcium ammonium nitrate

Principle

Ammonium nitrate or CAN sample is first distilled with weak alkali of MgO and
the released NH3 is absorbed in standard acid and estimated. Then the contents are
distilled with Devarda’s alloy and strong alkali of NaOH and the released NH3 is
absorbed in standard acid and estimated. Devarda’s alloy facilitates transformation of
nitrate form into ammoniacal form.

Reaction with MgO

2 NH4 (NO3) + Mg(OH)2 Mg(NO3)2 + 2 H2O + 2NH3
(NH4) + OH NH3 + H2O
2NH3 + H2SO4 (NH4)2SO4
With Devarda’s Alloy
Zn + 2NaOH Na2ZnO2 + 2H
NO3 + 9H NH3 + 3H2O

Reagents

1. Freshly ignited magnesium oxide

2. 0.1 N H2SO4

3. Devarda’s alloy

4. 40% NaOH

5. 0.1 N KOH

6. Methyl red indicator

7. Zinc pieces

8. Porcelain bits

113
Procedure

Weigh accurately 1 g of the NH4NO3 fertilizer, transfer it to a 250 ml volumetric

flask, dissolve in distilled water, and make up the volume and mix the solution
homogenously.

i) Estimation of ammoniacal N

Pipette out 25 ml of the above solution into the distillation flask. Dilute it with
400 ml of distilled water. Add a few pieces of zinc and porcelain bits. Pipette out 25 ml
of 0.1 N sulphuric acid into a 250 ml ice-tumbler or tall-form beaker. Add a few drops of
methyl red indicator and keep the standard acid under the delivery end of the distillation
unit with the tip immersed in the acid.

Then, add 10 g of freshly ignited MgO to the distillation flask, stopper it and
distill collecting the distillate in the standard acid container, till the distillate becomes free
of back-titrated ammonia. The ammonia in the distillate is absorbed in the standard acid
and the excess of acid is back-titrated against N /10 KOH. The ammoniacal N content in
the fertilizer is calculated from the volume of N / 10 acid utilized by the evolved
ammonia.

Calculations for NH4-N

Weight of fertilizer sample taken = 1.0 g

Volume made up = 250 ml

Volume of aliquot pipetted out for distillation = 25 ml

Volume of 0.1 N H2SO4 taken = X ml

Back titration value of N / 10 KOH = Y ml

? Actual volume of N/10 H2SO4 utilised by = X-Y = Z ml

the evolved ammonia

1 ml of N / 10 H2SO4 = 0.0014 g of N

? Z ml of N / 10 H2SO4 = 0.0014 x Z g of N

This is present in 25 ml of the aliquot

114
 In 250 ml of the fertilizer solution = 0.0014 x Z x 250
----- g of N
25

This is present in 1.0 g fertilizer sample

In 100 g fertilizer the amount of N = 0.0014 x Zx 250 x 100

----- ------
25 1

Estimation of nitrate N

After distilling the ammoniacal N present in the fertilizer solution, cool the
contents in the distillation flask. Add about 5 g of Devarda’s alloy and 10 ml of 40%
NaOH. Stopper the flask and continue the distillation, collecting the distillate in the
container with standard acid kept under the delivery end, till the distillate becomes free of
ammonia as tested with red litmus paper. Back-titrate the excess acid against standard
KOH and calculate the amount of NO3-N from the amount of standard acid utilized by
the evolved ammonia.

Calculations for NO3-N

Volume of N/10 H2SO4 utilized by the = Z mL

evolved ammonia
1 ml of N / 10 H2SO4 = 0.0014 g of N
z ml of N / 10 H2SO4 = 0.0014 x Z g of N
This is present in 25 ml of fertilizer aliquot =
? In 250 ml of fertilizer solution 0.0014 x Z x 250
----- g of N
25
This is present in 1 g of fertilizer sample
In 100 g fertilizer
ie., per cent of NO3-N in the fertilizer = 0.0014 x Zx 250 x 100
----- ------
25 1

115
 4.4 Estimation of Water- Soluble P in Superphosphate/Dicalcium Phosphate /
Rock Phosphate

Aim

To estimate the content of water-soluble P in superphosphate/ diammonium phosphate.

Principle

Phosphatic fertilizer is leached with distilled water through Whatman No.40 filter
paper. Phosphorous is then precipitated as ammonium phosphomolybdate in nitric acid
medium, filtered, washed free of acid and dissolved in a known excess of alkali and the
excess alkali is back titrated against nitric acid using phenolphthalein as indicator. From
the volume of alkali consumed, P content of the sample is arrived at.

Reaction during precipitation

2H3PO4 + 24(NH4)2MoO4 + 2 (NH4)3PO4.12MoO3 + 42 NH4NO3 +

42HNO3 4H2O
Reaction during dissolution of precipitate

2(NH4)2PO4.12MoO3 + 46 KOH 23 K2MoO4+ 2(NH4)2HPO4 + 22H2O

2(NH4)2HPO4 P2O5 + 4NH3 + 3H2O
Reaction during back titration

HNO3 + KOH* KNO3+ H2O

* Excess KOH not used in the dissolution of PO4 precipitate

From the equation it is seen that two molecules of (NH4)3 PO4.12 MoO3 will be
equivalent to one molecule of P2O5.

So, 46 molecules or 46 liters of 1N KOH = 142 g of P2O5

46 lit of N KOH = 142 g of P2O5
1 lit of N KOH = (142/46) g of P2O5 (or)
1000 ml of N KOH = (142/46)g of P2O5
1 ml of N/10 KOH = 0.000309 g of P2O5
1 ml of 0.1619 N KOH = 0.0005 g of P2O5

116
 The nitric acid used in back titrating the excess of standard alkali is of the same
strength as that of alkali i.e. 0.1619 N.

Reagents required

1. Conc. HNO3

2. 0.1619 N HNO3

3. 0.1619 N KOH

4. Ammonium nitrate

5. Phenolphthalein indicator

Procedure

Weigh accurately 1.0 g of superphosphate / DCP and transfer it to the funnel

fitted with Whatman No. 40 filter paper and collect the filtrate in a 250 ml volumetric
flask at the bottom. Leach the fertilizer with 10 ml portions of distilled water allowing
each portion to drain before adding the next portion and collecting the leachate in the 250
ml volumetric flask. Leaching must be continued all about 200 ml of leachate is collected
in the volumetric flask within one hour. Make-up the volume with distilled water after
adding a few drops of concentrated sulphuric acid if the leachate is turbid. Retain the
filter paper along with the residue for the estimation of citrate-soluble phosphorus.

Pipette out 25 ml of the extract into a 250 ml beaker and bring the solution to
nitric acid medium by adding ammonium hydroxide till the solution becomes alkaline to
litmus and then adding dilute nitric acid till the solution becomes acidic to litmus. Add
about 2 g of NH4NO3 and warm the contents to about 70oC on a water bath. Prepare the
precipitant mixture by adding 10 ml of 20% ammonium molybdate to 10 ml of 7:3 (7 ml
of conc. HNO3 + 3 ml of water) HNO3 in a 100 ml beaker. Add 20 ml of the precipitant
mixture slowly to the phosphate solution. Stir the contents gently without touching the
sides of the beaker. Keep the beaker on water bath at 70oC for half an hour and allow the
canary yellow precipitate to settle, leaving a supernatant solution.

Filter the supernatant solution through Whatman No.40 filter paper each time
transferring the supernatant solution only and retaining as much precipitate as possible in

117
the beaker. Give washings with cold water alternately to the filter paper and the
precipitate in the beaker using only 10 ml of water for each washing. Add the next
instalment of water to the filter paper only after the previously added water filtered
completely. Continue washings till the filtrate becomes free of acid (Collect half test
tube full of filtrate and add a drop of 0.1619 N KOH and a drop of phenolphthalein
Appearance of pink colour indicates free of acid).

After making the precipitate acid-free, transfer the filter paper along with the
precipitate to the same beaker in which precipitation was done and dissolve the yellow
precipitate by adding known excess of 0.1619 N KOH. Add 2-3 drops of
phenolphthalein and titrate against 0.1619 N HNO3. Calculate the amount of phosphorus
in the solution and in-turn in the fertilizer from the amount of 0.1619 N KOH utilized to
dissolve the ammonium phosphomolybdate precipitate.

Calculations

Weight of fertilizer sample taken = 1.0 g

Volume of the leachate made up = 250 ml
Aliquot of the leachate taken for estimation = 25 ml
Known excess 0.1619 N KOH added to the = X ml
precipitate
Volume of 0.1619 N HNO3 used in back = Y ml
titration
Volume of 0.1619 N KOH actually used to = X-Y = Z ml
dissolve the precipitate
1 ml of 0.1619 N KOH = 0.0005 g of P2O5
Z ml of 0.1619 N KOH = 0.0005 x Z g of P2O5
This is present in 25 ml of fertilizer leachate
In 250 ml of the leachate = 0.0005 x Z x 250
----- g of P2O5
25
This is present in 1 g of fertilizer
In 100 g SSP/DCP = 0.0005 x Z x 250 x 100
ie., % of P2O5 in the fertilizer ----- ------
25 1

118
 4.5 Estimation of Citric Acid-Soluble P in Dicalcium Phosphate / Rock Phosphate

Aim

To estimate the citric acid – soluble P content in the dicalcium phosphate / rock
phosphate sample.

Principle

The residue of the phosphate fertilizer remaining after leaching with distilled
water for water-soluble P estimation is extracted with ammonium citrate and the amount
of P in the extract is estimated by the volumetric method.

Reagents

1. 1% Ammonium Citrate

2. Conc. HNO3

3. Dil. HNO3

4. 0.1619 N HNO3

5. 0.1619 N KOH

6. 20% Ammonium Molybdate

7. Ammonium Nitrate

8. Ammonium Hydroxide

9. Phenolphthalein indicator

Procedure

Place the filter paper with the residue after leaching for water-soluble P
estimation, in a 500 ml conical flask and loosely stopper it. Add 100 ml of neutral
ammonium citrate solution of sp. gravity 1.09 and keep the flask on a thermostat at 65oC
for one hour, shaking the flask every 5 minutes. Remove the flask from the thermostat
and filter the contents immediately through a rapid filter paper (WhatmanNo.5). Wash the
residue on the filter paper with warm water (65oC). Collect the filtrate in a 250 ml
volumetric flask and make up the volume with distilled water. Estimate the citrate-
soluble P content in 25 ml of the extract by Pemberton method.
119
Calculations

Weight of fertilizer taken = 1g

Weight of the extract prepared = 250 ml

Volume of the extract taken for P estimation = 25 ml

Volume of 0.1619 N KOH added to dissolve = X ml

the precipitate

Volume of 0.1619 N HNO3 used in back = Y ml

titration

Volume of 0.1619 N KOH utilized to = X-Y = Z ml

dissolve the ammonium phosphomolybdate
precipitate

1 mL of 0.1619 N KOH = 0.0005 g of P2O5

Z mL of 0.1619 N KOH = 0.0005 x Z g of P2O5

This is present in 25 ml of aliquot

In 250 ml of the ammonium citrate extract 0.0005 x Z x 250

----- g of P2O5
25

This is present in 1 g of the fertilizer

In 100 g fertilizer 0.0005 x Z x 250 x 100

ie., per cent of citric acid soluble P2O5 in the ----- ------
fertilizer
25 1

(Note : 0.1 N KOH and 0.1 NHNO3 can also be used instead of 0.1619 N acid and alkali.
Then the factor for calculation will be 1 ml of 0.1 N KOH = 0.00039 g of P2O5)

120
 4.6 Estimation of Potassium in Potassium Chloride / Potassium Sulphate

Aim

To estimate the potassium content in the potassium chloride / potassium sulphate

fertilizer.

Principle

The potassium in the potassic fertilizer solution is measured in a flame

photometer. The flame photometry is based on the measurement of the intensity of light
radiation emitted by atoms on excitation when aspirated into a high-temperature flame.

Procedure

a) Preparation of fertilizer solution

Weigh accurately 1 g of KCl / K2SO4 sample, transfer it to a 100 ml volumetric

flask, dissolve in distilled water, make-up the volume and mix homogeneously. Pipette
out 1 ml of this solution into a 100 ml volumetric flask and make-up the volume with
distilled water and mix homogeneously.

b) Preparation of standard solution

Weigh accurately1.907 g of KCl after drying the salt at 105oC for 2 hours.
Transfer it to a 1000 ml volumetric flask, dissolve in distilled water make-up the volume
and mix homogenously. This gives the 1000 ppm K solution. Pipette out 25 ml of the
1000 ppm solution into 250 ml volumetric flask, dilute with distilled water, make-up the
volume and mix homogeneously. This gives the 100 ppm solution. Prepare standards
from 10 to 100 ppm by pipetting out 2.5, 5.0, 7.5, 10, 12.5, 15, 17.5, 20, 22.5 and 25 ml
of the 100 ppm solution into 25 ml volumetric flask and making up the volume of 25 ml.

a) Preparation of standard curve

Adjust the galvanometer or digital reading in the flame photometer to 0 and 100
by aspirating distilled water and 100 ppm K solution into the flame respectively. Then
aspirate the standard solutions from 10 to 100 ppm and record the meter readings. Draw
the standard curve with concentration K ppm on the x-axis and meter reading on the y
axis. A curvilinear curve will be formed.

121
d) Measurement of potassium in the fertilizer solution

Measure the potassium concentration in the fertilizer solution by aspirating the

solution into the flame of the meter and recording the meter reading. If the meter reading
crosses 100, the fertilizer solution may be diluted suitably. Using the meter reading the
concentration of K in the fertilizer solution may be read from the standard curve. Then
compute the potassium content in the fertilizer sample.

Calculations:

Weight of fertilizer taken = 1g

Volume of solution made-up = 100 ml

Volume of aliquot taken for dilution = 1 ml

Volume of the aliquot diluted = 100 ml

Concentration of K in the diluted fertilizer = X ppm

solution

% of K in the fertilizer = X 100 100

------ x ------ x ------- × 100
6
10 1 1

122
 4.7 Estimation of Nitrogen in Organic Manure / Oil Cake

Aim

To estimate the N content in the organic manure / oil cake.

Principle

Nitrogen present in organic form in the organic manure / oil cake is digested with
sulphuric acid containing salicylic acid in the presence of sodium thiosulphate and converted
to ammonium sulphate. Then the ammoniacal form of N is distilled and estimated.

Reactions

2NH4NO3 + H2SO4 → (NH4)2SO4 + 2HNO3

C6H4OH.COOH + HNO3 → C6H3OH.COOH.NO2 + H2O

Nitro salicylic acid

Na2S2O3 + H2SO4 → Na2SO4 + H2SO3 + S

C6H3OHCOOHNO2 + 3H2SO3 + H2O → C6H3NH2OHCOOH + 3H2SO4

2C6H3NH2OHCOOH + 27H2SO4 → (NH4)2SO4 + 26SO2 + 14 CO2 + 30 H2O

Reagents required

1. Conc. H2SO4

2. Salicylic acid

3. Sodium thiosulphate

4. Potassium sulphate

5. Copper sulphate

6. 40% NaOH

7. Sodium sulphide

8. N/ 10 H2SO4

9. N / 10 KOH

10. Methyl red indicator

123
Procedure

a) Digestion

Procedure

Weigh accurately 1 g of FYM/ compost / oil cake / green manure and transfer into
a macro Kjeldahl flask. Add 30 ml of conc. H2SO4 containing 1 g of salicylic acid. Mix
the contents and allow standing for at least half an hour with frequent shaking. Add
about 1.5 g of sodium thiosulphate (Na2S2O3.5H2O) and mix well. Digest over a low
flame until frothing ceases. Then add 10 g of K2SO4 and 1 g of CuSO4.5H2O and digest
the contents until the liquid in the flask turns apple green.

b) Distillation

Transfer the manure digest from the Kjeldahl flask into the distillation flask.
Rinse the Kjeldahl flask with distilled water 2-3 times and transfer the washings to the
distillation flask. Add 300 ml distilled water and a few pieces of metal zinc and porcelain
bits. Add 10 ml of 10% sodium sulphide and 120 ml of 40% NaOH. Stopper the flask
connecting with the distillation unit and distil the contents. Collect the distillate in a 250
ml ice tumbler / tall form beaker containing 25 ml of 0.1 N H2SO4 with methyl red
indicator. When all the ammonia is evolved as tested by red litmus paper at the end of
delivery tube, back titrate the excess acid against 0.1 N KOH. Calculate the percentage
of nitrogen in the organic manure /oil cake from the volume of 0.1 N H2SO4 utilized to
absorb the evolved ammonia.

Calculations

Weight of manure / oil cake taken = 1.0 g

Volume of 0.1 N H2SO4 taken in the ice-tumbler = X mL
Volume of 0.1 N KOH used on back titration = Y mL
Volume of 0.1 N H2SO4 utilized by the evolved ammonia = (X-Y) = Z mL
1 mL of 0.1 N H2SO4 = 0.0014 g of N
Z mL of 0.1 N H2SO4 = 0.0014 x Z g of N
This is present in 1 g of manure / oil cake
In 100 gm of manure/oilcake 0.0014 x Z x 100
i.e. % of N --------------------
1

124
 4.8 Estimation of Phosphorus in Organic Manure / Oil Cake

Aim

To estimate the P content in the organic manure / oil cake.

Principle

The organic manure / oil cake is digested in triple acid mixture of HNO3 + H2SO4
+ HClO4 in the ratio of 9: 2: 1. The digest is made upto known volume and the contents
of P in the extract are estimated calorimetrically.

Reagents

1. Triple acid mixture (HNO3 + H2SO4 + HClO4 in 9:2:1 ratio)

2. Barton’s reagent

3. Ammonium hydroxide

Procedure

a) Digestion

Weigh accurately 1 g of manure / oil cake and transfer it into a 100 ml of conical
flask. Add 12 ml of triple acid mixture and leave it overnight for cold digestion. Next
day digest the contents in a sand bath till the digest becomes colourless. Cool the
contents, rinse the sides of the flask and filter the contents through Whatman No.1 filter
paper and collect the filtrate in a 100 ml of volumetric flask. Wash the flask with
distilled water a few times and add the washings to the filter paper. Make up the filtrate in
the flask to volume and mix homogeneously. Estimate phosphorus in this extract.

b) Estimation of phosphorus

Pipette out 5 ml of triple acid extract into a 25 ml volumetric flasks. Add 5 ml of

Barton’s reagent (Barton’s reagent: Dissolve 6.25 g of ammonium molybdate in 100 ml
of warm distilled water and cool (Solution A). Dissolve 0.31 g ammonium meta vanadate
in 50 ml of hot water. Cool and add 62.5 ml of conc. HNO3 (Solution B). Add solution
A to solution B and make up the volume to 250 ml by adding 37.5 ml of distilled water).
Make up the volume to 25 ml mark, mix homogenously and allow for 30 minutes. Adjust
the meter reading to 100% of T/ OD by keeping the blank in the cuvette of a photoelectric
colorimeter. Now measure the % T/OD in the sample solution.

125
 Prepare a standard curve using P concentrations at 0, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6,
0.7, 0.8, 0.9, 1.0 ppm developing yellow colour as described previously and measuring
the % T or O.D. in the colorimeter. Draw the standard curve keeping the P
concentrations in the x axis and the % T or OD in the y axis, using this standard curve
find out the concentration of P in the sample solution and calculate percentage of P in the
manure / oil cake.

Calculations

Weight of manure / oil cake taken = 1.0 g

Volume of triple acid extract taken for P estimation = 5 ml

Volume of yellow colour developed solution made = 25 ml

up

Concentration of P in the sample solution = X ppm

Concentration of P in 25 ml solution in terms of = X x 25 g

micro gram

This is present in 5 ml of the triple acid extract

Amount of P in 100 ml of triple acid extract = (X x 25) x 100

----- g
This is present in 1 g of manure / oil cake 5

In 100 g of manure / oil cake, ie., per cent of P = (Xx 25) x 100 x 100
---------- ---- ----
6
10 5 1

126
 4.9 Estimation of Potassium in Organic Manure / Oil Cake

Aim

To estimate the K content in the organic manure / oil cake.

Principle

The organic manure / oil cake is digested in triple acid mixture of HNO3 + H2SO4
+ HClO4 in the ratio of 9: 2 : 1. The digest is made upto known volume and the contents
of K in the extract are estimated in a Flame Photometer.

Reagents

1. Triple acid mixture (HNO3 + H2SO4 + HClO4 in 9:2:1 ratio)

2. Ammonium hydroxide

3. 1000 ppm K

Procedure

a) Digestion

Weigh accurately 1 g of manure / oil cake and transfer it into a 100 ml of conical
flask. Add 12 ml of triple acid mixture and leave it overnight for cold digestion. Next
day digest the contents on a sand bath till the digest becomes colourless. Cool the
contents, rinse the sides of the flask and filter the contents through Whatman No.1 filter
paper and collect the filtrate in a 100 ml of volumetric flask. Wash the flask with
distilled water a few times and add the washings to the filter paper. Make up the filtrate in
the flask to volume and mix homogeneously. Estimate K in this extract.

b. Estimation of potassium in triple acid extract

Pipette out 5 ml of the triple acid extract into a 25 ml volumetric flask and
neutralize the acid by adding dilute ammonium hydroxide as tested by litmus paper.
Make up the volume to 25 ml mark and mix the solution homogenously. Measure the
potassium concentration in the sample solution flame photometrically.

Aspirate distilled water into the flame of the flame photometer and adjust the
galvanometer reading to zero. Aspirate 100 ppm K solution into the flame and adjust the

127
meter reading to 100. Now aspirate standard K solutions of 10, 20, 30, 40, 50, 60, 70, 80,
90 and 100 ppm and record the meter readings. Draw a standard curve, keeping the K
concentrations in the x axis and the meter readings in the y axis. Now measure the K
concentration of the sample solution using this standard curve. Calculate the percentage
of potassium in the manure / oil cake from the concentration of K in the sample solution.

Calculations

Weight of manure / oil cake taken = 1.0 g

Volume of triple acid made up = 100 ml

Volume of triple acid extract taken = 5 ml

Volume made up = 25 ml

Concentration of K in the sample solution = X ppm

Per cent of K in the manure / oil cake = ( X x 25) x 100 100

--- x ----
5 1

128
 REFERENCES

Chesnin, F. and C.H. Yien. 1951. Turbidimetric determination of available sulphates.

Proc. Soil Sci. Soc. Am., 18: 149-151.

Gupta, R.P and C. Dhakshinamoorthy, 1981. Procedure for physical analysis of soil and
collection of agrometerological data. Division of agricultural physics, IARI, New
Delhi.

Hesse, P.R. 1971. A text book of soil chemical analysis. John Murray, London.

Jackson, M.L.1973. Soil Chemical Analysis. Prentice Hall of India(P) Ltd. New Delhi.

Lindsay, W.L. and Norvell. 1978. Development of DTPA soil test for zinc, iron,
manganese and copper. Soil Sci. Soc. Am. J., 42: 421- 428.

Olsen, S.R. and F.S. Watanabe. 1957. A method to determine a phosphorus adsorption
maximum of soils as measured by the Langmuir isotherm. Soil Sci. Soc. Am.
Proc., 21:144-149.

Piper, C.S. 1966. Soil and Plant Analysis, Hans Publishers, Bombay, Monograph from
the Waits. Agricultural Research Institute, University of Adelaid., p.47-111, 197-
200.

Richards, I..A. 1954. Diagnosis and improvement of saline and alkali soils. USDA
Handbook No.60. pp160.

Sabula, R.A. 1973. Evaluation of analytical methods for determining K status of Nigerian
soils. K in tropical crops and soils. Proc.10thColloguium of the International
Potash Institute held in Dec.1973 in AbidjanIRepuplic of Ivory Coast-119-129.

Schoonover, W.R. 1952. Examination of soils for alkali. University of California.

Extension service, Berkeley, California.

Shoemaker, H.E., Mclean E.C and Pratt, P.F. 1961.Buffer methods for determining lime
requirement of soils with appreciable amounts of extractable aluminium. Soil Sci.
Soc. Amer. Proc. 25:274.

129
 Stanford, S. and L. English. 1949. Use of flame photometer in rapid soil tests of K and
Ca. Agronomy J., 41: 446- 447.

Subbiah, B.V. and C.L. Asija. 1956. A rapid procedure for the estimation of available
nitrogen in soils. Curr. Sci., 25:29-260.

Tandon, H.L.S. 1985. Methods of analysis of Soils, Plants, Waters and Fertilizers.
FOGO, New Delhi.

Tisdale, S. L. and W.L. Nelson. 1975. Soil Fertility and Fertilizers, 3rd ed. New York:
MacMillan Publishing Co., Inc. 694 p.

Truog, E.1960. Fifty years of Soil Testing. Trans 7th Int. Congr. Soil Sci.4:46-52.

Walkley, A. and C.A. Black. 1934. An examination of the Degtijareff method for
determining soil organic matter and proposed modification of the chromic acid
titration method. Soil Sci., 32: 233- 243.

Williams, C.H. and A. Steinbergs. 1959. Some soil sulphur fractions as chemical indices
of available sulphur. Australian J. Agric. Res., 10: 240-252.

130
 ANNEXURE-1

Safety Measures in the analytical laboratory

Analytical laboratory handles hazardous materials such as corrosive and toxic

chemicals besides many other radioactive materials. Apt handling during lab work is vital
for successful work output. Further, it avoids accidents within the lab by avoiding
mishandling. It averts health trouble associated with chemicals and corrosive materials
because of safe handling. In order to ensure the safety in analytical lab, the following
basic rules need to be adopted.

Basic rules for laboratory work

9 Always wear appropriate protective clothing (laboratory coat preferably of long

armed and knee length).

9 Wear appropriate footwear.

9 Use hand gloves whenever it is essential (at the time strong acid transfer).

9 Familiar with fire regulations in your laboratory and building.

9 Aware about the accident/emergency procedures in your laboratory and building.

9 Use appropriate glassware and devices for transferring liquids.

9 Use minimum quantity of chemicals required for the work.

9 Handling of hazardous chemicals should be strictly carried out within the fume hood
chamber.

9 Allow warm-up time for all instruments and check the functioning properly before
starting work.

Clean the working table, instrument and glassware immediately after work
completion.

9 Plan your work in advance and systematically progress as per work plan.

Do's in the laboratory

9 Wear chemical - resistant apron (lab coat) while working in laboratory.

131
9 Use volumetric flask for standard solution preparation.

9 Standardize acids and reagents before analysis.

9 Label reagent bottles properly with date along with mentioning the researcher who
has prepared.

9 Read instrumentation guide while handling new instruments.

9 Switch off instruments properly after work.

9 Enter log book in proper.

9 Read over the lab manual before starting experiment

9 Keep only essential lab notebooks, glassware and other items alone on the work table.

9 Never add water to acid as it will burst up. Always add acid to water and stir the
solution while adding the acid.

9 Keep glassware in concern tray safter completion of work.

9 Replace chemicals alphabetically after lab work.

9 Handle the instrument safter proper learning or with the lab technicians.

9 Carefully read the labels on chemical container before working with chemical.

9 Replace the lids or caps on bottles and jars after taking chemicals.

9 Put off Bunsen burner and gas cylinder immediately after completing heating works.

9 Use pyrex or quartz glassware alone for heating works.

9 Use minimum chemicals during analysis and avoid returning the excess chemicals to
the container (towards avoiding contamination).

9 Biological analysis should be carried out immediately.

9 Use suitable container or sheets for weighing chemicals and avoid placing chemicals
directly on the weighing pan.

9 Wash your hands with soap and water after each analysis.

9 Clean the acid and other chemicals spill up immediately in proper way.

132
9 Clean the work table, glassware and equipment properly after work completion.

9 After lab work, clean table and put glassware in washing tray immediately.

9 Discard acids if any in the glassware before putting for washing and was slightly with
tap water.

9 Before leaving lab ensures following.

9 Check water taps, water bath, distillation unit, light, fans, electrical points and gas
cylinder.

9 Switch off ovens, autoclave, computers, laminar flow and air conditioner.

Don'ts in the laboratory

9 Don't operate instruments without knowing how to operate or without lab technicians.

9 Don't pipette out acids.

9 Don't pour water in to acids.

9 Don't leave distilled water unit without anyone.

9 Don't work in UV light in Laminar Air Flow Chamber.

9 Don't use double distilled water for general chemical analysis.

9 Don't keep plant sample above 60°C in oven for drying.

9 Don't close windows in lab while working.

9 Don’t keep alcohol and acids near Bunsen burner/stove.

9 Don't leave gas cylinders in open condition.

9 Don't smell, touch or taste the chemicals.

9 Don't use non-standardized chemicals for analysis.

9 Don't runs soft electronic instruments while running heavy duty machineries like wily
mill, distillation unit, or any others.

9 Don't use the triple acid extract directly in Flame photometer. It will corrode the
burner. Mix with base towards neutralizing the acids.

133
9 Don't wear bulky or dangling clothing.

9 Don't run the kjelplus (Nitrogen) apparatus without reagents or distilled water in
appropriate container.

9 Don't smoke, eat or drink in the laboratory.

9 Place spectrophotometer cuvette in proper direction to avoid scratching in transparent sides.

9 Centrifuge should not be increased suddenly to top speed which causes failure of
centrifuge.

9 Never attempt unauthorized experiments

9 Never work alone in laboratory.

9 Finally don't hesitate to ask help. Lab In-charge is for facilitating you only

General safety measures

9 If chemical substances get in your eye, wash the eye out for 15 minutes. Hold your
eye open with your fingers while washing it out.

9 If you burn yourself on a hot object, immediately hold the burned area under cold
water for 15 minutes. Inform your teacher.

Burns

9 Small burns can be treated with 8% NaHCO3 for some time and then cover with zinc
oxide ointment.

9 Apply acriflavine or picric acid if any scalds caused by boiling water.

9 In case of acid burns, wash immediately with cold water and then with dilute 8%
NaHCO3 followed by application of acriflavine.

9 In case of caustic alkali burns, wash with water and then with 1% acetic acid.

9 In case of liquid Bromine burns, wash with petrol and apply olive oil.

9 In case of phosphorus burns, wash with cool water and then with silver nitrate solution.

134
Eye injuries

i) Acid in eye: Wash with water, then 1% NaHCO3

ii) Alkali in eye: Wash with water followed by I% boric acid

iii) Glass in eye: Remove the glass with forceps and call doctor immediately

Cuts

9 Minor cuts: Wash with 1% aqueous chloramines -T or 2% iodine solution. Remove

the glass, wash again, and apply sterilized dressing.

9 Serious cuts: Prevent bleeding and call doctor immediately

Learn proper handling techniques for all chemicals

1) Acids - Acids are highly corrosive, causing burns to skin and clothing.

2) Bases - Strong bases can also cause severe burns to the skin. Bases are very
dangerous if splashed into the eyes.

3) Mercury - Mercury is highly toxic and spills are difficult to clean up. If special
mercury recovery equipment is not available (sprays, wipes, etc. - available from
chemistry lab suppliers) zinc dust should be used. Zinc dust reacts with mercury to
form a safe mixture that is easy to handle and dispose of.

4) Dispose of all chemicals properly at the end of each laboratory exercise.

First aid box

First aid box should contain the following essential items towards handling
emergency health troubles in lab.

1) Bandages, lint, gauze, cotton, wool, adhesive plaster, etc.

2) Delicate forceps, needles, thread, safety pins, scissors.

3) Vaseline, salt, mustard powder, castor oil, olive oil, zinc oxide ointment, boric acid
powder, chloramines T (Hydrated crystals).

4) Lime water (0.04 N), 2% Iodine solution, 1% Boric acid, 1% acetic acid, 8%
NaHCO3, 1% NaHCO3, AgNO3, chloramines-T.

135
 ANNEXURE - 2

Common Terminologies

¾ Acid : A proton donor

¾ Aliquot: A portion of a solution

¾ Analysis sample: The actual size of the sample analysed

¾ Analysis: A process that provides chemical or physical information about the

constituents in the sample or the sample itself

¾ Analyte: A substance analyzed for its concentration

¾ Anode: The electrode where oxidation occurs

¾ Atomization: The process of converting the analyte into free atom

¾ Back titration: The reaction in which a reagent is added to a solution containing the
analyte and the excess reagent remaining after its reaction with the analyte is
determined by a titration

¾ Base: A proton acceptor

¾ Buffer: A solution containing a conjugate weak acid/weak base pair that is resistant
to a change in pH when a strong acid or base is added Cathode: The electrode where
reduction occurs

¾ Chromatography: A separation technique in which solutes partition between a

mobile phase and stationary phase

¾ Coagulation: The process in which smaller particles of precipitate clump together to

form larger particles

¾ Colorimetry is an analytical method based on comparison of colour intensities.

¾ Common ion effect: The solubility of an insoluble salt decreases when it is placed in
a solution already containing one of the salt's ions

¾ Complex formation reactions employ chelating or complex forming agents which

form bigger molecular weight complexes around the element or ion to be determined.

136
¾ Complexometry: A titration in which the reaction between the analyte and the titrant
is a complexation reaction

¾ Conductometry deals with the measurement of electrical conductivity and resistance

of a substance

¾ Coulometry is the measurement of current and time required for a chemical reaction.

¾ Desiccator: A closed chamber containing a desiccant used to store samples in a

moisture free environment

¾ Digestion: The process by which a precipitate is given time to form larger, pure crystals

¾ Dilution: The process of preparing a less concentrated solution from a more

concentrated solution.

¾ Displacement titration: A titration in which the analyte displaces a species from a

complex and the displaced species is determined by titration.

¾ Emission spectroscopy, the sample is subjected to an electric arc and the light
emitted is measured

¾ Emission: Release of a photon when an analyte returns to a lower energy state from a
higher energy state

¾ Enthalpy: The heat absorbed or released during a chemical reaction at constant pressure

¾ Entropy: A measure of disorder

¾ Equilibrium constant: It is the value that determines the relative concentrations of

products and reactants Equilibrium: A system is at equilibrium when the concentration of
the reactants and the products remain constant

¾ Flame Photometry, the sample is injected as a fine spray into the flame and the light
emitted is measured

¾ Fluorometry, commonly a metal fluorescent agent is excited by irradiation with

visible or UV radiation and the characteristic emitted radiation is measured.

¾ Gravimetry is the branch of analytical chemistry that takes into account of the
quantity of products of a reaction employed

137
¾ Gross sample: A sample which is collected from the target population without any
processing

¾ Kjeldahl analysis: An acid — base titrimetric method for determining the amount of
nobile nitrogen in organic compounds

¾ Laboratory sample: A homogenous portion of the gross sample taken into the lab
after form processing the gross sample

¾ Neutralization reactions are the types of analytical methods where reactions

between the acids and bases are involved. The determination of acids using a standard
base is in a acidimetry. If a standard acid is utilized for the determination of an
unknown base, the method is called alkalimetry.

¾ Occlusion: A co- precipitated impurity trapped within a precipitate form

¾ Ohm's law: The current flowing through a circuit is directly proportional to the
applied ant is potential and inversely proportional to the circuit resistance (E = iR)

¾ Potentiometry is the measurement of potential at an electrode, equilibrium with an

ion to be determined

¾ Precipitation methods are volumetric methods which result in formation of insoluble

precipitates. Ex. Argentimetry

¾ Tracer: A radioactive species used as an internal standard

¾ Turbidimetry and Nephelometry deals with the measurement of light stopped or

scattered by the precipitates

¾ Voltametry involves measurement of potential at micro electrode at a specific voltage

¾ Wavelength: The distance between two consecutive maxima or minima of an

electromagnetic wave

138
 ANNEXURE - 3

Equivalent weight of Substances used in volumetric Analysis

Valency/
S. Molecular Equivalent
Name of chemical basicity/
No. weight weight
acidity

1. Sodium carbonate (Na2CO3) 106.12 2 53.06

2. Sodium hydroxide (NaOH) 40 1 40.00

3. Oxalic acid (hydrated)( H2C2O4. 2H2O) 126.08 2 63.04

4. Hydrochloric acid (HCI) 35.46 1 35.46

5. Sulphuric acid (H2SO4) 98.08 2 49.04

6. Potassium permanganate (KMnO4) 158.05 5 31.61

7. Potassium dichromate (K2Cr2O7) 294.18 5 49.03

8. Ferrous ammonium sulphate (FeSO4 392.12 1 392.12

(NH4)2SO4 )

9. Iodine (I2) 126.92 1 126.92

10. Sodium thio sulphate (Na2S2O3.5H2O) 248.19 1 248.19

11. Copper sulphate (CuSO4 .5H2O) 249.68 1 249.68

12. Silver nitrate (AgNO3) 169.89 1 169.89

13. Sodium chloride (NaCl) 58.45 1 58.45

14. Potassium sulphocyanide (KCNS ) 97.17 1 97.17

139
 Molecular Molecular Equivalent
S.No Name of chemical
Formula weight weight

1. Succinic acid C4H6O4 118.00 59.00

2. Oxalic acid dihydrate C2OH2. 2H2O 126.00 63.00

3. Potassium hydrogen phthalate C8H6O4 204.22 204.22

4. Benzoic acid C6 H5COOH 122.22 122.22

5. Sulfamic acid NH2SO3 97.09 48.5

6. Sodium carbonate Na2CO3 106.00 53. 00

7. Borax Na2B4O7.10H2O 381.22 190.61

8. Sodium oxalate (COONa)2 133.99 66.99

9. Potassium dichromate K2Cr2O7 294.21 49.04

10. Arsenic trioxide As2O3 128.92 21.48

11. Sodium chloride NaCI 58.44 58.44

140
 ANNEXURE - 4

Guidelines for the Preparation of Standard Solutions

Weight / volume of substance
S.
Name of the chemical required for preparing 1 litre Standardise with
No.
0.1 N solution (g) or (ml)
1. Acetic acid 5.8 ml concentrated acetic acid 0.1 N KOH
2. Ammonium acetate 7.7 -
Ammonium flouride
3. 2.7
(NH4F)
4. Borax solution 1.907 -
(Na2B407.10H20)
5. Calcium solution 5.045 g CaCO3 -
0.1 N calcium
6. EDTA solution 10 g EDTA
solution
7. EDTA solution 18.6 g EDTA disodium salt 0.1 N calcium solution
8. Fehling's solution 69.28 g of pure CuSO4 in 1 litre 0.5 per cent Glucose
of water, 350 g of Rochelle salt
(Potassium sodium tartarate) and
100 g sodium hydroxide in 1 litre
of water
Ferrous ammonium
9. 39.2 0.1 N K2Cr2O7
sulphate
10. Ferrous sulphate 27.8 0.1 N K2Cr2O7
11. Hydrochloric acid 10 ml of conc. 11.3 N acid 0.1 N borax solution
12. Iodine Dissolve 20 g iodate free KI in -
30 to 40 ml of water, add 12.7 g
sublimed iodine and make up to
1 litre
13. Nitric acid 6.3 ml of conc. 16 N acid 0.1 N Na2CO3
14. Oxalic acid 6.296 -
15. Potassium carbonate 6.91 0.1 N HC1
16. Potassium chloride 7.45
17. Potassium dichoromate 4.904 Primary standard
18. Potassium hydroxide 5.7 to 6.0 0.1 N Succinic acid
Potassium hydrogen
19. 5.1 -
phthalate
20. Potassium permanganate 3.25 g 0.1 N oxalic acid or 0.1
N sodium oxalate
21. Potassium thiocyanate 10 0.1 N AgNO3

141
22. Silver nitrate 16.989 0.1 N NaCl

23. Sodium chloride (NaCI) 5.847

24. Sodium carbonate (Na2CO3) 5.29

Sodium bi carbonate
25. 8.401 0.1 N H2SO4
(NaHCO3)

26. Sodium hydroxide 4.0 0.1 N potassium

hydrogen phthalate

27. Sodium oxalate 6.69

28. Sodium thiosulphate 25.0 g Sodium thiosulphate. 0.1 N KMnO4 solution

Add 3.8 g of borax or 0.1 g or 0.1 N Potassium
of anhydrous Na2CO3 to iodate or 0.1 N iodine
stabilize the solution

29. Succinic acid 5.91 -

30. Sulphuric acid 3.0 ml of conc. 36 N acid 0.1 N Na2CO3

142
1. Bray No. 1 extractant 15 ml of 1N NH4F and 255 ml of 0.5N HCI are mixed
(0.03 M NH4F in 0.025 and the volume is made up to 500 ml with distilled
N HCl) water

2. Reagent A Dissolve 12 g of ammonium molybdate in 200 ml of

warm distilled water and cool. Dissolve 0.291 g of
antimony potassium tartarate in 100 ml distilled water.
Add both the solutions to 1000 ml of 5 N H2SO4 (138 ml
of con. H2SO4/1 water), mix homogeneously and make-
up to 2 litres with distilled water. Store in a pyrex glass
bottle in a dark and cool compartment. This solution is
heat and light sensitive.

3. Reagent B Dissolve 1.056 g of ascorbic acid in 200 ml of reagent

A. This reagent should be prepared freshly.

4. Conditioning reagent Dissolve 75 g 0 of NaCl in 275 ml distilled water in a

(turbid imetry) 500 ml volumetric flask and stir using a magnetic bar.
Add 30 ml of concentrated HCI, 100 ml of ethanol, and
50 ml of glycerol and stir until NaCl dissolves. Remove
stirring bar and makeup the volume with distilled water.

5. Standard Sulphate Dissolve 0.5434 g of AR grade K2SO4 in distilled water

in a 1 L volumetric flask and make up the volume to the
mark (100 ppm). Dilute 100 ml of this solution to 1 L to
obtain 10 ppm solution

6. 0.02 N EDTA 2 g dissolved in one litre water

7. DTPA Extractant DTPA extractant solution is prepared by dissolving 13.1

ml of TEA, 1.967 g of DTPA and 1.47 g of CaCl2 in 100
ml distilled water. The contents are allowed for some
time so that the DTPA will dissolve and then diluted to
about 900 ml. The pH of the solution is adjusted to 7.3 ±
0.05 with 1:1 HCI by stirring and the volume made upto
1 litre.

8. 0.5 M Sodium 42 g dissolved in one litre distilled water

bicarbonate

9. P standard solution Prepare 1000 ppm P solution by dissolving 0.4390 g of

AR grade KH2PO4 in distilled water and make up to 100
ml. Dilute 10 ml of this solution to 100 ml with distilled
water to prepare 100 ppm P solution. Dilute 10 ml of the 1

143
10. K standard solution Dissolve 1.907 of AR grade KCI in one liter of water.
This gives 1000 ppm K (1000 micro gram/ml). 100 ml
of 1000 ppm K is diluted to one litre which gives 100
ppm K. From this main stock solution, various standards
ranging from 10 to 100 ppm are prepared.

11. Zn standard solution 0.439 g ZnSO4.7H2O is dissolved in 200 ml distilled

water in a beaker to which 5 ml of 1:5 H2SO4 is added.
The contents are transferred to a one litre volumetric
flask and the volume made up. This gives 100 ppm of
Zn. From this stock solution working standards of 0, 0.1,
0.2, 0.3, 0.4 and 0.5 ppm solutions are prepared.

12. Fe standard solution 0.702 g ferrous ammonium sulphate dissolved along

with 5 ml of 1:5 H2SO4 and made up to 1 litre. This
gives 100 ppm of Fe stock solution. From this stock
solution various working standards viz., 0, I, 2, 3, 4 and
5 ppm are prepared.

13. Cu standard solution 0.392 g CuSO4.5H2O is dissolved in 1litre distilled water.

14. Mn standard solution 0.288 g potassium permanganate is dissolved in 300 ml

of distilled water. To this 20 ml of conc. H2SO4 is added
and warmed to 60°C. Then oxalic acid is added drop by
drop to make the solution colourless. The contents are
cooled and made up to 1 litre which gives 100 ppm
stock solution. From this, working standards of 0, 1, 2,
3, 4 and 5 ppm are prepared.

15. Saturated gypsum Five gram of pure gypsum is dissolved in one litre water
and kept overnight. On next day, shake well and filter it
out. This solution measures exactly 2.2 dS m-1

16. Ammonium chloride- 67.5 g of NH4C1 in 570 ml of NH4OH solution and the
Ammonium hydroxide volume is made up to one litre using distilled water
buffer solution

17. Buffer solution pH 4.0 Mix 50 ml of 0.2 M potassium thalate and 0.4 ml of 0.2
M NaOH and dilute to 200 ml

18. Buffer solution pH 7.0 Mix 50 ml of 0.2 M monopotassium phasphate and

29.54 ml of 0.2 M NaOH and dilute to 200 ml

19. Buffer solution pH 9.2 0.5 M borax solution

144
 Approximate Volume required to
make 1000 ml of
S. No. Name
Specific Per cent approximately 0.1 N
Normality
gravity by weight solution (ml)

1 HCl 1.18 35 11.3 8.9

2 HNO3 1.42 70 16.0 6.3

3 H2SO4 1.84 96 36.0 2.8

4 H3PO4 1.69 85 41.1 2.3

5 CH3COOH 1.05 99.5 17.4 5.8

6 Aqueous NH3 0.90 27 14.3 7.1

7 H2O 1.00 100 - -

145
 ANNEXURE - 5
Guidelines for the Preparation of Indicator Solutions

S.No Name of the indicator Preparation

1 Methyl orange Dissolve 0.5 g of the saltin 1 litre distilled water and f
(CI4H15N3SO4)
2 Methyl Red (C15H15N30) Dissolve 0.5 g of methyl red in 100 ml of 95% alcohol
3 Phenolphthalein Dissolve 0.5 of phenolphthalein in 100 ml of 95%
(C2OH14O4) alcohol
4 Bromocresol green — Dissolve 5 g of bromocresol green and 1 g of
Methyl red (Double methyl red in 100 nil of 95% ethanol and neutralize
indicator) to mid colour (bluish purple) with a few drops of N,
NaOH (The indicator is pink at pH 4.2 or below and
bluish green as the pH rises to 4.9 and above).
5 Potassium chromate Dissolve 5.0 g AR potassium chromate in 100 ml of
(K2CrO4) distilled water
6 Diphenylamine Dissolve 0.1 g of diphenyl amine in 10 ml conc.
[(C6H5)2NH] H2SO4
7 Potassium ferricyanide Wash out the superficial coating of ferrocyanide
[(K3Fe (CN)6] with distilled water and soak it in a filter paper.
Now weigh 0.1 g of this washed crystal and
dissolve in 100 ml distilled water.
8 Dissolve one gram of bromocresol green in 100 ml
Bromocresol green
alcohol
9 Grind 0.5 g of ammonium purpurate in 100 g of
Ammonium purpurate
K2SO4
10 Eriochrome Black T Dissolve I g in 100 ml alcohol
11 Universal indicator Dissolve 0.05 g methyl orange, 0.15 g methyl red,
0.30 g bromothymol blue and 0.35 g phenolphthalein
in 66% alcohol and make upto 1 litre with alcohol
12 Murexide 0.2 g of ammonium purporate, 10 g of ammonium
chloride and 40 g of potassium sulphate are mixed
and ground well.
13 Starch indicator I% solution of soluble starch chemical in water.
Boil the content for homogeneous dissolutions. 2 g
of boric acid can be used as a preservative to arrest
the conversion to sucrose

146

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