Preface This book can be considered as an enlarged and revised edition of my earlier book “Soil Sampling and Methods of Analysis”. The earlier edition was widely accepted and found to be useful to the students, teachers and researchers working in this field. By this time, I received lot of suggestions and requests from the readers for its improvement and inclusion of few more chapters related to this field. Keeping these views in mind, both publisher and author have agreed upon to publish this book as a new one wherein 8 new chapters have been included. This book is comprised of 27 chapters besides references and 16 appendices at the end. Each chapter is ended with some relevant questions and answers on that topic for the benefit of the students. In addition to ‘Soil Physical and Chemical Analysis’ this book covers two more major aspects namely ‘Soil Mineralogical Analysis’and ‘PlantAnalysis’.The typographical errors committed inadvertently in the earlier book have been taken care of here. Hope, like earlier one, this book will definitely fulfil the need of the students, teachers and researchers engaged in the field of agriculture, geology, earth science, etc. Like past, any sort of suggestions from the readers for its improvement in future will highly be appreciated. The author gratefully acknowledges the suggestions received from the readers of all corners of this country and abroad and each source from where the technical matters are drawn.

Soil is a mass of large variability and thus its sampling is the most challenging problem for any soil analyst. Practically, soil sampling is a two step process which includes (i) collection of soil from homogeneous unit and (ii) processing which in turn includes drying, grinding, sieving, mixing, partitioning and storing.

2.1 Particle Size Distribution Analysis Soil is composed of inorganic particles and organic molecules of varying size and shape. The expression of the proportion of the different size inorganic particles is termed as ‘soil texture’. It is one of the most important basic characters being little changed by cultivation or other practices. Particles are arbitrarily distinguished into following groups on the basis of their diameters.

3.1 Determination of Bulk Density of Soil 3.1.1 Determination of Bulk Density of an undisturbed Soil Bulk density of soil is expressed as the ratio of the mass (weight) of soil particle to their total volume including the pore space between the soil particles. Alternately, it is the weight of unit volume of dry soil. It is usually expressed in the unit of gram per cubic centimeter (g/cc) or mega gram per cubic meter (Mg/m3). In fine textured soil bulk density varies from 1.00 to 1.60 g/cc, whereas in coarse textured soil from 1.20 to 1.80 g/cc.

4.1 Determination of Specific Surface Area of Soil Specific surface area of a soil is defined as surface area per unit weight of soil and is expressed as m2 g-1. The specific surface of soil greatly influences the physical and chemical properties such as retention of water at high suction, swelling, plasticity, soil strength, cation exchange capacity and availability of nutrients. The basic principle of determination of specific surface is based on the amount of gas or polar liquid required to form a monomolecular layer on the soild. Knowledge of molecular size (diameter) and the mass of the adsorbate enables one to calculate specific surface of the adsorbent. Principle Dyal and Hendricks (1950) introduced a method for estimating specific area based upon the adsorption of ethylene glycol (EG) to form a monomolecular layer over the entire soil surface. The method involves adding EG to pretreated soil or clay sample and evaporating the excess EG in an evacuated system. The quantity of EG retained at the moment the evaporating rate decreases is proportional to the surface area. Later on Carter et al. (1965) suggested the use of ethylene glycol monoethyl ether (EGME) having high vapour pressure at room temperature than EG for determining surface area of the layer silicate minerals and soils.

In natural condition primary soil particles cohere to each other to form a cluster or an aggregate through the cementing action of various agents like organic matter, carbonates, oxides of iron and aluminium, etc. Aggregate analysis tells one about the structural status, pore geometry which in turn influences the movement of air and water in soil, amount of water stable secondary particles (aggregate) as well as the extent of finer particles aggregated to the coarser particles and the erodibility status of surface soil against wind and water.

6.1 Determination of Soil Moisture 6.1.1 Determination of Soil Moisture Content by Gravimetric Method (Direct method) Traditionally, water content in soil is expressed as the ratio of the weight of water present in the soil to the weight of dry soil. When this ratio is multiplied by 100, it becomes the percentage of water in the soil sample on dry weight basis. Equipments and Materials Soil auger, aluminium moisture box with air tight lid, balance, oven and desiccator

7.1 Determination of Hydraulic Conductivity of Saturated Soil Laboratory Methods Hydraulic conductivity of a saturated soil indicates the ease with which water is transmitted through the soil pores. If a constant water head (h) is maintained on a saturated soil column of length (L) and the volume of water (V) percolating per unit time (t) through per unit cross sectional area (A) to the other end, then according to Darcy’s Law, the rate of flow of liquid or flux (q) is proportional to the hydraulic gradient (ÄH/L) across the length of soil column.

Soil consistency is a term used to designate the manifestation of different physical forces acting within the soil at various moisture levels. The consistency limits, also known as ‘Atterberg Limits’ (Atterberg, 1911; 12) are defined by the water content of the soil to create specified degree of consistency. These limits are – (i) Liquid limit or upper plastic limit, (ii) Plastic limit or lower plastic limit and (iii) Sticky limit. 8.1 Determination of Liquid Limit or Upper Plastic Limit Equipments and Materials Liquid limit device (Casagrande, 1932) and accessories, spatula, dish, oven, moisture box, balance, desiccator

9.1 Determination of Oxygen Diffusion rate (ODR) of Soil The rate of movement of oxygen from atmosphere to respiring plant root cells is important for root respiration and thereby the growth and development of plant. While diffusion through the air-filled pores maintains the exchange of gases between the atmosphere and soil, diffusion through water films maintain the supply of oxygen and disposal of carbon dioxide from active root tissues which are typically hydrated. Principle Lemon and Erickson (1952) introduced a method for measuring oxygen diffusion in soil with the help of platinum electrode. The process measures the extent of chemical reduction of oxygen by the platinum electrode maintaining a constant potential of 0.65V with a reference electrode.

Temperature is a fundamental property of usually used to characterize the thermal condition of a system. The growth of any biological system is optimal within certain ranges of temperature and inhibited or ceased beyond this boundary. In addition to plant growth, the significance of temperature on agriculture can be ascertained by its role on physical, chemical and biological processes occurring in soil. Therefore, to study these soil properties intensively measurement of soil temperature is essential. 10.1 Measurement of Soil Temperature Temperature of any system cannot be directly measured. It can only be estimated by its influence on some properties (thermometric properties) which respond to the variation in the intensity of heat of that system. These thermometric properties include volume, pressure, length, electrical resistance, thermal emf of the matter. Instruments built to take the advantage of any of these thermometric properties of the matter are called thermometer.

11.1 Determination of Penetrability of Soil Penetrability of a soil is usually assessed by the penetration resistance, which a measure of soil strength and thereby the soil compaction. Penetration resistance is the capacity of the soil in its confined state to resist penetration by a rigid object. Besides soil strength, the penetration resistance also depends on the size, shape and orientation of the axis of the penetrating probe. The penetrating probe may be a finger, pencil, stick, root or any specially designed object having a specified geometric shape and a device to measure the resistance as it is pushed into the soil. Soil penetrability is intimately related to agriculture because of its adverse impact on draught requirement during ploughing, root development and crop yield, to civil engineering due to its relation to settlement, stability and ground water flow. Soil penetrability is a measure of the ease with which an object can be pushed or driven into the soil. Any device designed to measure the resistance to penetration may be called penetrometer. There are several types of penetrometers: (i) pocket penetrometer, (ii) proctor penetrometer, (iii) cone penetrometer, and (iv) split-spoon penetrometer. Here, we will discuss about two most commonly used penetrometers viz. pocket penetrometer and cone penetrometer used for agricultural purposes.

Thermal Analysis Physical and chemical properties of any soil are governed largely by the mineralogical make up of the soil, and more specifically by the clay fraction. Identification, characterization and an understanding of properties of different minerals are important in evaluating soils in relation to classification, agronomic practices and engineering purposes. Clay fractions of soils are usually composed of mixture of one or more secondary phyllosilicate minerals along with primary minerals inherited from the parent materials. Identification of minerals species as well as their quantitative estimation in such polycomponent system require application of several qualitative and quantitative analysis. The methods used for identification and quantification have steadily been improved over the last 50 years. Among the several methods often used for this purpose three are very common, e.g. (i) differential thermal analysis (DTA), (ii) x-ray diffraction analysis (XRD), and (iii) infrared spectroscopy (IR). Thermal analysis is a term covering a group of analyses that determine the changes of some physical parameters, such as weight, energy, dimension and evolved volatile substances with change in temperature. Many soil constituents undergo various thermal reactions upon heating, which can be utilized as a diagnostic tool for qualitative identification and quantitative estimation of the substances. These reactions may be exothermic or endothermic, and phase change or crystal change may take place during reactions. Whatever be the reactions, if properly measured, they give some valuable information to identify the substance under consideration. Among the several methods available for thermal analysis, three methods are frequently used in soil research: differential thermal analysis (DTA), differential scan calorimetry (DSC), and thermogravimetry (TG). Among these, DTA and TG will be discussed here.

The total energy of a molecule consists of translation, vibrational, rotational and electronic energies. Transition among different types of energy levels occurs in different regions of electromagnetic spectrum. Transitions between rotational and vibrational energy levels of the ground state molecules give rise to absorption bands throughout the infrared region of the electromagnetic spectrum. Organic molecules are characterized by possessing bonds and groups that vibrate independently of each other. Thus, initially infrared spectrometry was used for identification of functional groups of organic molecules. There is lot of ambiguity in the interpretation of infrared spectra of organic matter preparations due to complexity and diversity of the components of soil organic matter. But if used carefully it can provide information on: (i) nature, reactivity and structural arrangement of oxygen-containing functional groups, (ii) presence of protein and carbohydrate constituents, (iii) presence or absence of inorganic impurities (metallic ions, clays, etc in humic fractions), (iv) the amount of different soil organic matter components, and (v) interaction of organic components with pesticides, herbicides, fertilizers, etc. On the contrary to organic molecules, minerals tend to have few isolated vibrating groups. It can be used for mineralogical analysis when used in conjunction with x-ray diffraction and other similar techniques. Infrared spectrometry is used in the identification of inorganic compounds including minerals which have well-defined absorption bands in determining- (i) whetherlayer silicate isdioctahedral or trioctahedral in composition, (ii) in studying isomorphous substitution, (iii) hydration of minerals, and (iv) in quantitative analysis. Infrared spectrometer provides information about nature and identify of inorganic compounds that are amorphous in x-ray diffraction analysis.

14.1 Principles of X-ray Diffraction Any crystalline structure is characterized by a regular and systematic arrangement of atoms (or ions) in a three-dimensional array. Since crystals are composed of regularly spaced atoms or ions, each crystal contains planes of atoms or ions separated by a constant distance which is the characteristic of that crystalline species. X-rays are essentially electrostatic and electromagnetic fields oscillating in planes perpendicular to each other and to the direction of propagation in periodic cycles. X-rays having wavelengths in the order of 10-3 to 101 nm, are produced within a vacuum x-ray tube by bombardment of a metal anode with high-velocity electrons. High-velocity electrons transfer their energy to the electrons of metal anode causing their elevation to higher energy level (excited state). Excited electrons of metal atom soon come back to the vacant shell and each electron transfer from higher to lower energy state results release of a quantum of energy (x-ray photon) equivalent to the difference in energy between these two levels. The energy and the wavelength of emitted x-ray photons are finite and characteristic of the particular atoms of the metal anode used in x-ray tube. Usually, photons generated due to electron transfer from L-shell to K-shell are used for x-ray diffraction analysis. Diffraction of x-ray involves scattering of x-rays by planes of atoms of the crystal separated by definite distance and reinforcement of scattered rays in definite direction away from the crystal. Reinforcement of scattered rays is quantitatively related to the distance of atomic planes in a crystal as defined by Bragg’s equation:

15.1 Law of Mass Action Gulberg and Waage, the two Norwegian chemists in the year 1864 stated the law of mass action in this form: ‘The velocity of any chemical reaction at any instant at a given temperature is proportional to the active mass of each of the reactants at that instant present in the system, the active mass in a homogeneous system is defined as the number of gram molecule of the substance present per unit volume i.e., the molar volume’.

16.1 Balance and Weighing 16.1.1 Principle of Functioning of a Balance It is one of the important tools used in the laboratory and one should have thorough knowledge aboutitsassembly, use and care,otherwise it may be the initialstep for introducing error in any physical or chemical analysis. Every balancehas its maximum load limit and sensitivity depending upon its construction. Essentially balance may be regarded as a rigid beam, BC having a central fulcrum, O and two arms of equal length, d1, d2 (Fig.16.1). Theboth ends of beam have prism edges upon which the balance pans are suspendedby means of suitable suspension.

17.1 Determination of Soil pH The soil pH value is the measure of hydrogen or hydroxyl ion activity in the soil solution and it indicates whether the soil is acidic, neutral or alkaline in reaction. The pH is the most important property of soil as it controls the availability of plant nutrients, microbial activity and physical condition of soil. Since crop growth is affected both under very low (strongly acidic) and very high (strongly alkaline) soil pH, reclamation of these soils are necessary. Soil pH in fact indicates the extent of active acidity (activity of hydrogen ion in soil solution) of soil. Depending upon the purpose of measurement and soil condition, soil pH is measured in several soil-water or soil-salt solution ratios. The saturated paste is used for identifying specific soil problems like degree of acidity or alkalinity. For making fertilizer recommendation, 1: 2 soil-water suspension is generally adopted. To mask the variability in salt content of the soil, the pH is measured in soil-CaCl2 suspension (1:2 soil-0.1 M CaCl2 solution). The soil pH value is independent of dilution over a wide range of soil-salt solution ratio, thus more reproducible than those obtained with soil-water suspension.

18.1 Determination of Electrical Conductivity of Soil Solution The conductivity or more precisely the specific conductance of a soil at a given soil-water ratio is the reciprocal of the specific resistance offered by the solution at 25ºC between the electrodes of 1 sq cm cross section kept 1 cm apart. Thus, the electrical conductivity of any solution depends on the amount of soluble salt present in it. Knowledge on the extent of salinity is important for management of saline soil including the choice of the crop suitable for that soil. Soluble salt content in saturation extract is more reliable index of salinity level than in soil: water ratio of 1: 2 or 1: 2.5, because it simulates the extent of salinity experienced by crop in field moisture condition. Salinity class of soil is based on the electrical conductivity (EC) of saturation extract at 25ºC. However, EC of water extract from 1:2 soil-water suspension can be transformed into the EC of saturation extract following the formula (except for soil containing gypsum).

Carbon may be present in soil in four different forms: Carbonates: Mainly as CaCO3 and MgCO3, active CO3 2- and HCO3 - ions and gaseous CO2. Highly condensed elemental organic form: Charcoal, graphite. Completely decomposed resistant organic residues: Humus. Undecomposed and partially decomposed organic residues. The latter three fractions constitute the total organic carbon of which last two are chemically active form.

20.1 Exchange Capacity Soil colloidal particles carry both negative and positive charges on their surface which attract and hold equivalent amount of oppositely charged ions to their surface. The total number of negative charges present on a given mass of soil or capacity of a given mass of soil to exchange cations or total number of cation held by a given mass of soil is termed as cation exchange capacity (CEC). Similarly, the capacity of a soil to retain anions is known as anion exchange capacity (AEC). The ion exchange capacity of a soil depends upon the type and amount of colloidal particle (clay and organic matter) present in the soil. Ion exchange capacity (CEC/AEC) is expressed as milliequivalent ion (cation/ anion, respectively) per 100 g of soil (meq/100g). It is also expressed as centimoles of charge (positive/negative) per kg of soil [cmol (±) / kg]. Meq/ 100g = cmol (±) / kg of soil. 20.1.1 Determination of Cation Exchange Capacity of Soil Principle Soil is first saturated with normal ammonium acetate (pH 7.0). Excess ammonium (NH4+) ions and displaced cations are then removed by washing with alcohol. The amount of NH4+ ions retained by the soil is measured by steam distillation of NH4-saturated soil with MgO. During distillation evolved ammonia is absorbed in a boric acid solution as ammonium borate and the amount ammonium borate formed is determined by titration with standard sulphuric acid in presence of mixed indicator.

21.1 Determination of Total Nitrogen in Soil (Modified Kjeldahl Method) Nitrogen mostly present in soil in organic form. Relatively small amount of nitrogen usually occurs in ammonium and nitrate form, the available form. The Kjeldahl method of total N determination includes both organic and ammonium forms and with modification nitrate form of N can also be included. Principle When soil is digested with concentrated sulphuric acid organic nitrogen is converted to ammonium sulphate. To speed up the reaction a digestion mixture composed of sodium (or potassium) sulphate (to raise the boiling point of H2SO4) and cupric sulphate (CuSO4) - selenium powder (catalyst) is used.

Phosphorus is present in soil both in inorganic form and organic form. The inorganic P can be divided into different pools. The major active inorganic forms are: P bound to aluminium (Al-P), iron (Fe-P), calcium (Ca-P) and silicate minerals and relatively less active forms are occluded and reductant soluble forms of P. On the other hand, the principal organic P compounds present in soil are (i) inositol phosphate, (ii)phospholipids, (iii)nucleicacid and other unidentified esters and phosphoproteins. 22.1 Determination of Available Soil P The availability of soil P to plant varies greatly depending upon the reaction, minerological composition and amount of the soil colloid. Very little P is present in the soil solution except in recently fertilized or sodic soils. The type of phosphate ions present in the soil solution depends on the soil pH. In soils of neutral to slightly alkaline pH, HPO42-ion is the dominant form. In slightly or moderately acidic soils, both HPO-and HPO2-ions prevail. At strongly acidic soil HPO 244 24 ion tends to dominate, while above pH 9.0, the PO43-ion becomes more important - than H2PO4 . As the P is drawn from soil solution largely through diffusion by the plant, it is replenished from solid phase pool of available P. P fertilizer is added to recharge this pool.

23.1 Forms of Soil K Ignoring potassium (K) in soilorganism, total K in soilmaybe classified as (i) water soluble K, (ii) readily exchangeable K (exchangeable K), (iii) slowly exchangeable K (non- exchangeable or fixedK) and (iv) mineral or lattice K. Thedifferentforms are in dynamic equilibrium with one another.

Sulphur (S) occurs in soil in both organic and inorganic forms. Inorganic S fraction may occurs as sulphate (SO42-) and compounds of lower oxidation state such as sulphide (S2-), polysulphide (Sn2-), thiosulphate (S2O32-) and elemental sulphur (S0). In well drained soils inorganic S usually present as sulphate. Under anaerobic condition (reduced or waterlogged soil) inorganic S mainly occur as sulphide and under extreme condition as elemental S form. Estimation of S is carried out in two steps: Extraction of different forms of S from soil by suitable extractant. The filtrate is then analysed for S by the turbidimetric method. The turbidimetric method is the most widely used one because of its simplicity and rapidity.

25.1 Determination of Available Micronutrient cations (Zn, Cu, Fe and Mn) in Soil The available micronutrient cations (Zn, Cu, Fe and Mn) can be estimated in a single extraction with diethylene triamine pentaacetic acid (DTPA) as proposed by Lindsay and Norvell (1978). DTPA has the excellent property to combine with free metal ions in the solution forming soluble complexes. The stability constants of metal-DTPA complexes make DTPA as a most suitable extractant for simultaneous complex formation with Zn, Cu, Fe and Mn. In calcareous soil DTPAextractant may result overestimation due to excess dissolution of CaCO3, causingrelease of occluded micronutrient cations which are not plant available. To avoid this excessive dissolution of CaCO3, DTPA is buffered at pH 7.3 using triethanol amine (TEA). At this pH major part (3/4th) of TEA is present as HTEA+ (protonation) and exchanges for Ca and Mg ion from the soil exchange site. This increases concentration of Ca and Mg in the solution phase and suppresses dissolution of CaCO3. The DTPA has the capacity to form complex with each of the micronutrient cations 10 times of its atomic weight and it ranges from 550 to 650 ppm depending on the micronutrient cation. The amount of cation in the extract is determined on an atomic absorption spectrophotometer (AAS). The experimental condition such as shaking time, DTPA concentration, pH and temperature during shaking influence the amount of Zn, Cu, Fe and Mn extracted. Increase in shaking time and temperature markedly influence the extractability of these cations.

26.1 Plant Available Fraction of Nickel, Cadmium, Lead and Chromium The DTPA method was originally developed to extract Mn, Cu, Fe and Zn from slightly acidic to alkaline soils (Lindsay and Norvell, 1978), but in some cases it can be used for other metals also, such as Nickel (Ni), Cadmium (Cd), Lead (Pb) and Chromium (Cr). Among them Ni is considered as an essential micronutrient. The DTPA is used to extract soil metals by forming chelates with free metals in solution, which lowers their ionic activities so that additional quantities of metal are released from the soil colloids until equilibrium is attained. Triehanolamine (TEA) is used to buffer the extracting solution at pH 7.3 in an attempt to optimize the extraction of various metals. Calcium chloride is included in the extracting solution to minimize dissolution of CaCO3.

Different diagnostic tools are designed to avert nutrient deficiency or sufficiency and if properly used both loss of yield and quality of crop can be minimized. Among several diagnostic tools, nutrient analysis of standing crop seems to be an efficient method in arriving at the need-based fertilizer scheduling for various crops. Nutrient uptake by plants indicates the actual accessibility of nutrients in soil to the plants. Also, concentration of certain heavy metals in plant indicates their solubility in soil which leads to poor crop quality.Although, different plant species and even different varieties of same species may differ in their nutrient requirements, the concentration of nutrient element in plant can act as guide to support the soil test result in assessing nutrient supplying capacity of a soil and as a tool for correcting any deficiency if carried out early enough to prevent yield loss. However, analysis of harvested or mature crop is like a post mortem as this information only can help in nutrient management planning in subsequent years. Like soil testingplant analysis also suffers from some limitations. Amongthem, the most important are the risk of sampling error as well as the chances of incorrect interpretation of test report in recommending fertilizers. Selection of right plant part and right stage of crop growth are very crucial for successful plant analysis.

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