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IIMTERACTIOIM OF ORGANIC POLLUTANTS AND TRACE METALS WITH CROPS AND

SOILS OF ALIGARH DISTRICT

RESUME

THESIS SUBMITTED FOR THE DEGREE OF

Bottor of $litla£iopI)p IN

CHEMISTRY

BY

JAMAb AHMAD KHAN

DEPARTMENT OF CHEMISTRY ALIGARH MUSLIM UNIVERSITY

ALIGARH (INDIA)

2000

< < ^ n a Az:ul i;;^<> ' • ^ ' .

-ru,r^7^555'6 V' i ( Ace. No......

^sume

In recent years, a voluminous amount of work has been reported on the

use of organic chemicals, sewage sludge, industrial wastes, etc. to agricultural

lands for boosting crop output. However, indiscriminate and ever-increasing

application of these materials has culminated in inducing toxic effects on plants,

humans and other non-target organisms. The present study entitled "Interaction

of organic pollutants and trace metals with crops and soils of Aligarh

district" enumerates the effects of amino acids, pesticides, surfactants, organic

acids and fly ash on physico-chemical properties of soils of Aligarh district.

Aligarh, constituting a part of the Uttar Pradesh, lies to the North of the

Ganga-Yamuna Doab. It has a geographical ar'ea of 5000 sq. km. (including

newly created Hathras district) and lies between latitude of 27° 29' and 28° 11'

north and longitude 77° 29' and 78° 38' east. Soils of the Aligarh district are

alluvial with little leaching and considerable accumulation on the surface.

According to the order of geneses of the principal soil types, the district of Aligarh

has been divided into six typical soil zones, which was selected for study.

This thesis has been divided into following five chapters along with

general introduction:

Chapter I: Mobility of amino acids through soil amended with some

pesticides.

Chapter II: Effect of fly ash on the mobility of amino acids through six

typical soils of Aligarh district.

Chapter III: Mobility of trace metals in soil as influenced by some

surfactants.

Chapter IV: Effect of fly ash on the physico-chemical properties of soil and

seed germination, growth and metals uptake of barley and

wheat plants.

Chapter V: Effect of endosulfan on the seed germination, growth and

nutrients uptake of fenugreek plant.

A concise account of the results achieved on the basis of the plan,

mentioned above, is presented as:

General Introduction covers the survey of past and present literature with

particular references to the composition of soil, soil microorganisms and their

activities, physico-chemical properties of soil, soil adsorption, soils of India,

general description of Aligarh soils, plant nutrients, soil thin-layer

chromatography, amino acids, pesticides, surfactants, organic acids, trace

elements and their effects, fly ash and soil pollution.

Chapter I deals the mobility in terms of Rf values of some amino acids, viz.

lysine, glycine, alanine, phenylamine and valine through soil amended with some

pesticides such as parathionmethyl, dimethoate, carbendazim, chlorpyrifos,

endodulfan and methyldemeton by using soil thin-layer chromatography. It was

observed from the results, that the mobility of amino acids followed the order:

valine> alanine> phenylalnine > glycine in natural soil. The amendment of soil

carried out by pesticides, has shown the variation in the mobility of amino acids.

The mobility of valine, alanine, and lysine was found to decrease where as, that

of phenylalanine was increased on increasing concentration of pesticides (up to

100 mg 100g' soil). However, an enhanced mobility of glycine was noticed up to

75 and 50 mg lOOg' soil, respectively for parathionmethyl and dimethoate and

for carbendazim, chiropyrifos, endosulfan and methyldemeton. The results have

been explained on the basis of the nature and chemical behaviour of amino acids

in soil medium (pH 8.5).

Chapter II describes the effect of various doses of fly ash (0, 5, 10, 15 and

20 g kg' soil) on the mobility of amino acids viz. lysine, glycine, alanine and

valine through six principal soil types of Aligarh district viz. Ganga Loamy Sand

(Type I), Aligarh Loam (Type II), Aligarh Clay Loam (Type III), Aligarh Sandy

Loam (Type IV), Yamuna Silty Clay Loam (Type V) and Yamuna Sandy Loam

(Type VI). The results showed that the mobility of amino acids followed the order:

valine > alanine > glycince > lysine in each soil. This mobility trend showed a

variation in different soil types and followed the order: Type I > Type IV > Type II

> Type VI > Type III > Type V, which is related to the porosity of soils posed by

sand-silt contents and inversely related to their clay contents. The mobility of

lysine was found to decrease on increasing concentration of fly ash while, a

decreasing trend has been noticed in case of glycine, alanine and valine at low

initial concentration but it increased at higher concentration. The results are

explained on the basis of chemical nature of amino acids, fly ash and soils.

Chapter III reports the mobility of Pb, Cu, Ag, Zn and Co trace metals in

soil as influenced by CTAB, Triton X-100 and SDS surfactants, using distilled

water and organic acids (formic, acetic and oxalic) solutions as mobile phase.

Results showed that the mobility was found in the order: Co > Zn > Ag > Cu > Pb

through natural soil. However, the mobility of these metals was found to increase

in case of CTAB and Triton X-100 throughout the entire range of amendments.

On the other hand, the mobility was found to decrease in SDS-soil system. The

results have been explained on the basis of the nature and chemical behaviour of

surfactants and metal ions in the soil medium.

Chapter IV deals with the effect of fly ash (0,5,10,15,20,25,30 and 35 g

Kg' soil) on the physico-chemical properties of the soil and seed germination,

growth and metals uptake of barely {Hordeum vulgare L.) and wheat {Triticum

aestivum L.) plants. The beneficial effect on the physiological parameters was

noticed at lower doses of fly ash (up to 30 and 26 g Kg' soil in case of barely

and wheat, respectively), thereafter, a phytotoxic behaviour was observed. The

analysis of plants showed the increase in the concentration of K, Mg, Fe and Zn

metals up to 30 and 20 g fly ash Kg'"" soil in barely and wheat, respectively,

thereafter, they tended to decline on increasing doses of fly ash. On the other

hand, the uptake of Na, Cr, Mn, Cu, Ni, Cd and Pb contents was found to remain

enhanced throughout the entire range of fly ash amendments. The variation in

physico-chemical properties of soil, due to the addition of fly ash, has indicated a

decrease in pH (from 8.50-7.95) and composition of sand and clay and an

increase in E.G. and composition of silt and organic matter. The results have

been explained on the basis of composition and behaviour of fly ash.

Chapter V describes the effect of endosulfan on physiological parameters

such as seed germination, shoot and root length, number of leaves and nutrients

uptake of fenugreek {Trigonella foenumgraecum L). The results have indicated

that endosulfan was phytotoxic even at its lower concentrations. The absorption

of certain nutrient elements such as Ca, Mn, Co and Cu was facilitated at lower

doses (up to 500 mg kg' soil) and thereafter the inhibition of nutrients absorption

was noticed. However, the inhibitory trend for Mg, P, K, Fe and Zn uptake was

observed throughout the entire range of endosulfan amendments. The results

have been explained on the basis of toxic behaviour of endosulfan on the plant

and soil microbes.

INTERACT/OW OF ORGANIC POLLUTANTS AND TRACE METALS WITH CROPS AND


/" j ^

THESIS SUBMITTED FOR THE DEGREE OF

Bottor of $i)ilQ£(opt)p IN

CHEMISTRY

BY

JAMAb AHMAD KHAN

DEPARTMENT OF CHEMISTRY ALIGARH MUSLIM UNIVERSITY

ALIGARH (INDIA)

2000

ALIGARH MUSLIM UNIVERSITY AllGAkH - 202 002

CERTIFICATE

This is to certify that the work embodied in this thesis entitled "Studies on

the interaction of organic pollutants and trace metals with crops and soils of

Aligarh districV^ is the original work done by Mr. Jamal Ahmad Kh?n under

our supervision and is suitable for submission for the degree of Doctor of

Philosophy in Chemistry.

(Prof. SamiullaltAhan) Co-supervisor

Department of Applied Chemistry

Faculty of Engineering and Technology

a#Res: (091-0571)701881

Off: (091-0571)700920-21

Ext. 456

(ProfAhafiullah) Supervisor

Department of Chemistry

S # Res: (091-0571) 502247

Off: (091-0571)700920-21

Ext: 318

DEDICATED TO

MY FATHER AND

TO THE LOVING MEMORIES OF MY

LATE MOTHER AND

LATE SISTER

Acknowledgement

lAJiln the utmoit faith in the LJmniicient, the LJmnipreient

ana the LJmnipotent, J/ place tnii record of- m^ reiearcn activitiei in

tne nanai o / tnu Cearnea exaininen. J^ nave preientea here Ike oeit J '

coil Co. ana am futiu re dp on ii Lie for ik o minor errord ana oiniddiond t/iat

minlit nave inaavertentcu crept in ipite of- utmoit caref-ulneA6.

3t ii inaeea a matter of- areat pleaiure for me to expreii mu

indeliteaneii for tne liountiei -^ nave l)een tne tieneficiaru of ou virtue of

oeina a ituaent of tnid dream floiver of îr Jûed ^^nm,ad ^J\lian. -^

owe tnuc h to m^ ^tmamater.

J-^rofeiior ^nafiuccafi, department of i^nemiitru, mu mentor and

auide, had tjeen a areat iource of hnowledae and inipiration to me all

alona tfie duration of- tliii worh. w/ tlian/i him for tiii cliaracteriiti

u/illin^neii in aivina me a lot of time from nii ouiu icnedule and ft

hii aenuine affection towardd me.

~y am immeniet^ tlianhful to m^ co-auide, f-^rofeiior âmiuitak

^J\nan, deparltnenl of .Applied i^nemiilru, whoie icnotarlu advice and

guidance Itas t>een tne pillar of mu itrenatn.

J/ owe everuiiting cfratitude to f-^rofeiior âeeduzzafar,

chairman, department of L-kemiitri^ and f-^rof. ^Js.Cf. Uardkneu,

cliairman, department of Applied L^kemiitr^, for tkeir periiitent

ic

or

intet'eit and aeneroui ketp in prouidinq reiearcn (acititied. fr/v tnanhi

are alio due to ex-ckairtnen, prof. flarJ JiLm, frof. W- Jiâi,

Prof. J(.W. SLn^Suddin, prof W. ^^nâi and prof JJ.S.

iKatkor for prouidinq reiearck and laboratoru facilities durinq ikeir

respective periods.

Wit/i fond memoru, ^ remember oLate f-^rof. III. .J^kakaouddin

-J4kinad for enliqkteninq me u/itfi liis Sound advice and Suqqestions.

J7 would like to tliank f-^rof-. III. Iiluskfiq. and oDr.

.Îiamskuzzaman for tkeir vaiualjle suqqestions. ~y also u/ant to express

mu qratitude to all teackers and reSearck SckolarS of tke department of

L^kemistru and^^pplied L^ketnistru for extendinq full cooperation and

encouraqement durinq mu researck studies.

^y sincerelq acknowledqe witk tkanks tke vaiuaole Suqqestions of

mq seniors ^Jr. Kao LJwais Ât. ^\kan, oDr. Aamai Zriroz, Ujr.

-Atfreen J^ttamitn an d Wirs. SLeL. Wlq appreciation is also due to

mq lab coUeaques Iflr. Jâlman, Iffr. Kakesk, Jjr. (Lkteskam,

Wrs. SLeeL Skafi, W . . lâLkanda, % . Skaqufta, Wji.

Vazisk, 7//lr. ^ftak and Wr. Wekkook.

w/ wisii to express mq keartfelt and sincere tkanks to cJjr.

ôkrab ^\lian, oZ)r. J^kafiutlak and I fir. Kakesk.

J / am particularlq indebted to mq friends oZ)r. irjokd. ^sif

Wluskarraf and Wr. Wokd ^kram J<kan and kostei feiLws Wr.

•uzzaman, /f/r. Zreroz ^J4aiaer, -Atauocate an

Sraridi for their continued moral Aupport, nelp and encouragement

tfirouafiout tliis dtudu.

a dpeciai aedture of tnu deep appreciation ana reaard j-or my,

^y^ppi, tJjr. 2Janira ^\lian, J/ extend my tnanhi to tier for Iter

affectionate beliauior and encouraaement at all leveli durina tfiii itudu.

^ inall fje retniie in moral dutiei if ^ do not expreii mu feelinai

of qratitude for tlie multifarioui contributions of mu uncle, If jr.

moodur r\e/iman, and aunty, âmeela, towards Supporting me in

facina tnanii odds durina tlie u/fiole of trie tenure of my study, ''ly

aratefuinesi is also due to <UJr. _Xr. >A/. ^^nsari and oDr. ^^leem

^\ltan for tlieir Auaaestions.

JJne painstakina iob of typina by iVlr. -Jrrif Aamal Sîddiqui

and il/lr.ÂtbidAamalîddiqui is nianly appreciated.

~yn tlie last, but not certainly the least, ^ feel immense pleasure

in expressing the deepest aratilude to my parents and otlier family

tnembers for their everlastina moral Support, affection and patience

without which it would have been impossible for me to accomptisn tlie

arduous tash of completinq this th lesis.

Contents Page Nos.

LIST OF FIGURES (i)

LIST OF TABLES (ii-iii)

LIST OF ABBREVIATIONS (iv-v)

LIST OF PUBLICATIONS (vi)

GENERAL INTRODUCTION: 1-82

CHAPTER I: MOBILITY OF AMINO ACIDS THROUGH

SOIL AMENDED WITH SOME PESTICIDES

INTRODUCTION 83-84

EXPERIMENTAL 85-96

RESULTS AND DISCUSSION 97-101

REFERENCES 102-103

CHAPTER I I : EFFECT OF FLY ASH ON THE MOBILITY

OF AMINO ACIDS THROUGH SIX TYPICAL


INTRODUCTION 104-105

EXPERIMENTAL 106-i17


REFERENCES 121-122

CHAPTER llhMOBILITY OF TRACE METALS IN SOIL AS

INFLUENCED BY SOME SURFACTANTS


EXPERIMENTAL 125-135


REFERENCES 139-140

CHAPTER IV:EFFECT OF FLY ASH ON THE PHYSICO-

CHEMICAL PROPERTIES QF SOIL AND

SEED GERMINATION, GROWTH AND

METALS UPTAKE OF BARLEY AND

WHEAT PLANTS




REFERENCES 158-160

CHAPTER V: EFFECT OF ENOOSULFAN ON THE SEED

GERMINATION, GROWTH AND NUTRI

ENTS UPTAKEOF FENUGREEK PLANT




REFERENCES 173-174

( i )

LIST OF FIGURES

Figure Page Nos.

1 Schematic sketch of crystal structure of montmbrillonite clay

mineral 4

2 Layer structure of montmorillonite clay mineral 5

3 Schematic sketch of crystal structure of kaolinite and illite clay

minerals 7

4 Layer structures of kaolinite and illite clay minerals 8

5 Soil map of Aligarh district 29

6 Mobility of amino acids through soil amended with some

pesticides. 95

7 Effect of fly ash on the mobility of amino acids through six

typical soils of Aligarh district 116

8 Mobility of trace metals in surfactants amended soil 128

9 Effect of formic acid on the mobility of trace metals in

surfactants amended soil 130

10 Effect of acetic acid on the mobility of trace metals in


11 Effect of oxalic acid on the mobility of trace metals in


12 Effects of fly ash on the growth of barley plant 148

13 Effects of fly ash on the growth of wheat plant 149

14 Effects of fly ash on the seed germination and growth of

barley plant 150

15 Effect of fly ash on the seed germination and growth of wheat

plant 151

16 Effect of endosulfan on the growth of fenugreek plant 165

17 Effect of endosulfan on the seed germination and growth of

fenugreek plant 166

( i i )

LIST OF TABLES

Table P^Q® "os .

I Some general characteristics of Type I (Ganga Khadar)

* and Type II (Eastern Uplands) soils of Aligarh district 31

II Some general characteristics of Type III (Central Lowland)

and Type IV (West Uplands) soils of Aligarh district 32

III Some general characteristics of Type V (Yamuna Khadar)

and Type VI (Trans Yamuna Khandar) soils of Aligarh

district 33

IV Physico-chemical properties of Hirapur fine sandy loam soil 92

V Mobility of amino acids through soil amended with some

pesticides 96

VI Structures and characteristics of amino acids 98

VII Some physico-chemical properties of Aligarh soils 111

VIM Physico-chemical properties of fly ash 112

IX Effect of fly ash on the mobility of amino acids through six

soil types of Aligarh district 117

X Mobility of trace metals in surfactants amended soil 129

XI Effect of formic acid on the mobility of trace metals in


XII Effect of acetic acid on the mobility of trace metals in


XIII Effect of oxalic acid on the mobility of trace metals in


XIV pKa values of organic acids and solubility of trace metal

(Hi)

carboxilates and hydroxides 138

XV Concentration of available metals in soil and fly ash 145

XVI Physico-chemical properties of fly ash amended soil 146

XVII Physiological parameters of barley and wheat grown on fly

ash amended soil 152

XVIII Effect of fly ash on the metals uptake of barley 154

XIX Effect of fly ash on the metals uptake of wheat 155

XX Physico-chemical properties of soil 164

XXI Effect of endosulfan on the seed germination and growth of

fenugreek plant 167

XXII Effect of endosulfan on the nutrients uptake of fenugreek

plant 170

(iv)

LIST OF ABBREVIATIONS

Ml

<

>

°c AP

BSS

Cmol

CEC

CTAB

dSm"''.

DTPA

E

EC

EDTA

FA

FAQ

Fig.

g

ha

HP

Hrs.

J&K

LUP

m ha

meq

mg

Mins.

ml

Micro-liter

Less than

Greater than

Degree centigrade

Andhra Pradesh

British Standard Sieve

Cent! mol

Cation exchange capacity

Cetyltrimethylammoniun bromide

Desi siemen per meter

Diethylenetriaminepentaaciticacid

East

Electrical conductivity

Ethylenediamic tetracetic acid

Fly ash

Food and Agricultural Organization

Figure

Gram

Hectare

Himachal Pradesh

Hours

Jammu and Kashmir

Land use planning

Million hectare

Milli equivalent

Milligram

Minutes

Milliliter

(V)

mm Millimeter

mmho millimho

MP Madhya Pradesh

N North

NBSS National Bureau of Soil Survey

NH4OAC Ammonium acetate

pH Potentia hydrogenii

pi Isoelectric point

ppm Parts per million (w/w)

Rf Migration distance relative to solvent front

SDS Sodiumdodecyl sulphate

Sec. Second

Sq. km. Square kilometer

TEA Triethanolamine

TLC Thin-layer chromatography

TN Tamil Nadu

UP Uttar Pradesh

USDA United States Department of Agriculture

w/v Weight / volume

w/w Weight / weight

WB West Bengal

( v i )

LIST €P DIJBLICATI€N$

1. S.U. Khan; Shafiullah and Jamal A. Khan. Mobility of amino acids through Soil amended with some pesticides. Indian J. Environ.; 41(2): 83-89, 1999.

2. S.U. Khan; Jamal A. Khan; R.K. Bhardwaj and Jabir Singh. Influence of Ni(ll) and Cr(lll)-humic acid (HA) complexes on the available major nutrients status (NPK) of the soil, Poll Res.; 18(4): 511-514(1999).

3. S.U. Khan; Shagufta Jabin; R.K. Bhardwaj and Jamal A. Khan. Effect of organic compounds on the mobility of trace metals through soil amended with fly ash. Indian J. Chem. Soc; 77: 326-328 (1999).

4. S.U. Khan; R.K. Bhardwaj, Shagufta Jabin and Jamal A. Khan. Influence of chemical fertilizer on the mobility of heavy metals through soil amended with fly ash. Poll. Res.; 1999 (Accepted).

5. S.U. Khan; Shagufta Jabin and Jamal A. Khan. Influenced of some pesticides on the mobility of some trace metals through soil amended with and without fly ash. Poll. Res.; 2000 (accepted).

6. S.U. Khan; Jamal A. Khan; Shagufta Jabin and R.K. Bharadwaj. Mobility of trace metals in soil as influenced by some surfactants. Clay Res.; 2000 (accepted).

7. S.U. Khan; Jamal A. Khan and Jabir Singh. Effect of fly ash on the growth. Development and metal ions uptake by pigeon pea {Cajanuscajan), black gram {Phaseolus mungo) and okra {Abelmoschus) plants. India J. Environ. HLTH; 1998 (Communicated).

8. S.U. Khan; Shafiullah and Jamal A. Khan. Influence of organic acids on the mobility of heavy metals in soil amended with some insecticides. India J. Environ. HLTH; 1998 (Communicated).

9. S.U. Khan; Jamal A. Khan and Shafiullah. Effect of fly ash on the mobility of amino acids soils through six typical soils of Aligarh district. Chromatographic Science; 1999 (Communicated).

10. S.U. Khan; Jamal A. Khan and Shagufta Jabin. Effect of fly ash on the physico-chemical properties of soil and seed germination growth, and metals uptake of barley and wheat. Eco. Environ. Cons.; 2000 (Communicated).

11. S.U. Khan; Jamal A. Khan and Shagufta Jabin. Effect of endosulfan on the seed germination, growth and nutrients uptake of fenugreek plant. Industrial Poll. Control; 2000 (Communicated).

GENERAL

INTRODUCTION

QeneraC introduction

C3y» he term soil is derived from Latin word "Solum" which

C / means floor. According to Hilgard (1892), "Soil is more or

less a loose and friable material in which plants, by means

of their roots find a foothold for nourishment as well as for other

conditions of growth". This definition emphasizes the soil as a medium

for crop growth, which is not always true because there are some soils

(highly sodic or severely acidic), which may not be capable of producing

anything; nevertheless they are soils.

According to the glossary of soil science terms (Soil Science

Society of America, 1970), "Soil is (i) the unconsolidated mineral

material on the immediate surface of the earth that serves as a natural

medium for the growth of land plants, (ii) the unconsolidated mineral

matter on the surface of the earth that has been subjected to and

influenced by genetic and environmental factors of parent material,

climate (including moisture and temperature effects), macro- and

microorganism and topography, all acting over a period of time and

grsm'RM' im9m>vcn(XN'

producing a product that is soil, that differs from the material from

which, it is derived in many physical, chemical, biological and

morphological properties and characteristics".

COMPOSITION OF SOIL

Soil is mainly composed of solid, liquid and gaseous phases in

varying proportion. The approximate composition of an ideal surface

soil is given below:

Solid (50 %)

Inorganic

Organic

Sand Primary minerals

[ (quartz, micas, feldspares)

Clay Secondary minerals (clay minerals hydrous oxides)

Humus

Liquid (25 %)

Gaseous (25 %)

Solid phase:

Composition of solid phase of soil comprises (i) inorganic

materials and (ii) organic matter.

gfEsm^^M- iJfPRsyDVcTKm 3 1

(i) Inorganic materials:

The constituent of inorganic materials in soils is composed of

small rock fragments and minerals of various kinds, which have been

formed by physical and chemical weathering. They are extremely

variable in size. The smallest constituent of materials is clay

(<0.002mm), which is the most active and reactive portion of soil

(Baver, 1948). The smallest clay particle has colloidal properties (Grim,

1953; Mackenzie, 1957; Olphen, 1963; Marshal, 1964) and is composed

of few simple building units. Some important and most frequently

occurring clay minerals in soil are:

Montmorillonlte:

Montmorillonite has the 2:1-type (Fig. 1 and 2) mineral layers,

which is made up of an octahedral (alumina) sheet sandwiched between

two tetrahedral (silica) sheets (Hofmann et al., 1983). These layers are

loosely held together by very weak oxygen-to-oxygen and cation-to-

oxygen linkage. Exchangeable cations and associated water molecules

are attracted between the interlayer spaces, causing expansion of the

crystal lattice. Montmorillonite exhibit high plasticity, cohesion and

marked swelling when wet and shrinkage on drying. Cation exchange

capacity of montmorillonite is high, that varies from 60 to 150 Cmol (p+) •,t)0

per kg (or meq g"^)soil (Daji, 1990).

Kaolinite:

Koalinite is the most prominent member of 1:1-type (Fig. 3 and 4)

crystal among the isomers, halloyslte, nacrite and dickite. The layers of

SILICON TETRAHEDRAL SHEET

Crystal unit

14 A (9.6-10 A) or more

I

^SSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSS SILICON TETRAHEDRAL SHEET ^

O Oxygen linkage

O Oxygen linkage

ssss ™ ^ SILICON TETRAHEDRAL SHEET ^ ^

b-Axis

Crystal unit

Interlayer space

* S ^ SILICON TETRAHEDRAL SHEET ^ ^ ^ ^ ^ ^ ^

Crystal unit

Montmorillonite (2:1-type expanding minerals)

Schematic sketch of crystal structure of montmorillonite clay mineral

Fig. 1

o o o o

'W'^t^^o^^xr TETRAHEDRAL SILICON SHEET

J(g ))L (S l J© ^^®<^ ^ S i j ©

5

OCTAHEDRAL GIBBSITE SHEET

nH 0 B

EXCHANGEABLE CATIONS

UA

Tetrahedral sheet

c-Axis

Octahedral sheet

Tetrahedral sheet

£x^^^h^^s>^^: o (Si, Al)

•12

+16 or less

4 O +2 (OH) -10

4 (Al, Fe, Mg) +12 or less

4 O +2 (OH) "lO

b-Axis

Montmorillonite (2:1-type expanding minerals)

Layer structure of montmorillonite clay mineral

4 Si

6 O

Net Charge

+16

-12 0 or less e.g -1

Fig. 2

gw^<Rjic im^cxDVcitcm

silicate are made up of one tetrahedral (silica) sheet combined with one

octahedral (alumina) sheet (Pauling, 1930; Ross and Kerr, 1931;

Grunar, 1932). The tetrahedral and octahedral sheets in a layer of

kaolinite crystal are held together tightly by oxygen atoms, which are

mutually shared by the silicon and aluminium cations in their respective

sheets. These layers in turn, are held together by hydrogen bonding. In

contrast with other silicate groups, kaolinite exhibit very little plasticity,

cohesion, shrinkage and swelling. It has restricted surface and limited

adsorptive capacity for cations and water molecules. The cation exchange

capacity is very low and varies from 5 to 15 Cmol (p+) per kg soil (Daji,

1990).

Iinte:

lllite has 2:1-type (Fig. 3 and 4) mineral layers and is very similar

to that of montmorillonite structure. The major source of charge is due

to the isomorphous replacement of ions in the tetrahedral sheet (Grim

et al., 1937; Bolt et ai, 1963) where about 20% of the silicon sites are

occupied by aluminum atoms. This result in the balance of net negative

charge of the tetrahedral sheet. To satisfy this charge, potassium ions

are strongly attracted in the interlayer space that act as a bridging

agent, preventing the expansion of the crystal layers. Cation exchange

capacity of illite is 20 to 40 Cmol (p+) per kg soil.

ALUMINIUM OCTAHEDRAL SHEET

Crystal Unit

Interlayer Space

b-AXIS

Kaolinite (1:1-type mineral)

Crystal Unit

(10 A)

i

^ SILICON TETRAHEDRAL SHEET

yK

SILICON TETRAHEDRAL SHEET

O Oxygen linkage

:yK

b-AXIS-lllite (2:1-type non-expanding minerals)

^ Crystal Unit

Interlayer Space

Crystal Unit

^ ^ SILICON TETRAHEDRAL SHEET ^ ^

Schematic sketch of crystal structure of kaolinite and illite clay minerals

c-Axis

Octahedral sheet

Tetrahedral sheet

J

t < ^ . .

2 A ;

0"

^

L a

T T T T T Hydrogen bonding

(^.. M , . ' © - . @>«,'@ ' <°"' •« ^:'^:; " > : ; :::'-; ; : K ; 4 AI +12

Q ' (§) ^ P ' 0 ' © ^ 0+2 (OH) -10

J. b-Axis

Kaolinite (1:1- type mine

IL ^ ^ 4 Si +16

^ 3 ^ 6 0 -12 Net charge Q

ral)

yK yK yK yK

loA Tetrahedral sheet

c-Axis

Octahedral sheet

Tetrahedral sheet

^ ^ ^ j N ^ ^ r ^ ^

b-Axis

lllite (2:1-type expanding mineral)

Layer structures of kaolinite and illite clay minerals

deficiency charge is balanved by K ions

6 0

4 (Si.AI)

4 0 +2 (OH)

4 (AI.Fe, Mg)

4 0 + 2 (OH)

4 (Si.AI)

6 0 Net charge

-12

+16 or less

-10

+12 or less

-10

+16 or less

-12

0 or less e.g. -1

gw^vu: um^ptrnJcnom

Chlorite:

Chlorite is basically iron-magnesium silicate associated with

some aluminium (Donahue et al., 1977). In a typical chlorite clay

crystal, 2:1-type layers are formed. The crystal unit contains two silica

tetrahedral sheets and two magnesium-dominated trioctahedral sheets,

giving rise to the term 2:1:1 or 2:2-type structure. There is no water

adsorption between the chlorite crystal units, which accounts for the

non-expanding nature of this mineral. Chlorite has low cation exchange

capacity and varies from 10 to 40 Cmol {p+) per kg soil.

Amorphous clay:

Amorphous clay is the mixture of silica and alumina, containing

weathered oxides of iron etc. that has not formed very well oriented

crystals. These are common in soils and exist in varying proportions.

Their properties are uncommon usually shown by well-defined clay. On

the contrary, they posses some times positive charge, which is

responsible for anion exchange property, which is governed by the pH

and the nature of salt solution in soil medium.

(ii) Organic matter:

Soil organic matter comprises an accumulation of partially

disintegrated and decomposed plant and animal residues and other

organic compounds synthesized by the soil microbes. They are also

gE!m<RM- i7fism>'0cTi(m 10

called humus. Organic matter binds the mineral particles into granules

that are largely responsible for the loose and easily manageable

condition of the productive soils with good water holding capacity. The

humus is usually dark black or brown in colour and is colloidal in

nature. It posses high cation exchange properties and go up to 350

Cmol (p+) per kg soil. Humus may be divided into following main

groups:

Humic substances:

Transformed products bearing little or no resemblance to the

anatomical structures from which they are derived.

Non-humic substances:

Unaltered remains of plant and animal tissues and also organic

compounds that have definite characteristics.

According to their solubility in alkali and acids, the humic

substances can further be sub-divided into three sub-groups.

Humic acids: Soluble in alkali and insoluble in acids.

Fulvic acids: Soluble in both alkali and acids.

Humin: Insoluble in both alkali and acids.

Accumulation patterns of humic and fulvic acid fractions have

been studied by various workers (Syers et al., 1970; Schnitzer and

griym^ijiL im^RptDVcncm ii

Khan, 1972; Goh et al., 1976; Kilham and Alexander, 1984) to show

that the humic acid gets accumulated principally in the surface of

immediately below the surface horizon while fulvic acid fraction extends

to much lower depth and is hardly precipitated from its alkaline extract

by acid because of the presence of more functional groups (Stevenson

and Goh, 1971) in the structure.

Liquid phase:

It is mainly composed of soil water. Soil water is an important

component of soil. It has the property of solublizing the plant nutrients

in their usable or ionic forms (Brady, 1974). The osmotic pressure on

the hair like roots is also controlled by the quantity of water or salt

concentration. Soil water, therefore, becomes one of the limiting factors

in crop production.

The total water present in the soil is not utilized by plants. The

water that can be absorbed by the plant is termed as available water

and that water, which is not absorbed by the plant, is termed as non-

available water.

Gaseous phase:

Composition of gaseous phase in soil is soil air, which is dynamic

and varies greatly from place to place within a given soil (Russel and

Appleyard, 1915; Jong, 1981). It contains a number of gases, of which

g<E3FE<RM' irniRp^vcncm 12

nitrogen, oxygen, carbon dioxide and water vapour are tiie most

important constituents. The soil air contains a much greater proportion

of carbon dioxide and lesser amount of oxygen than atmospheric air.

This is due to respiratory activity of the roots, soil flora and fauna. Soil

air is essential for the respiration of soil microorganisms and

underground parts of the plants.

The content and composition of soil air are determined largely by

the water content of the soil, since the air occupies those soil pores,

which are not filled up with water.

SOIL MICROORGANISMS AND THEIR ACTIVITIES

The various microorganism, that inhibit the soil forms are one of

the important components of all soils. It has been estimated that one

gram of soil may contain 1 to 100 million microorganisms, forming a

very small fraction (<1%) of soil mass (Donahaue et al., 1977; Sehgal,

1996). They are considered to be present in soil from the top to a depth

of few inches. The density of microbial population and their normal

activities depend on many factors, such as the availability of food and

energy supply, moisture, temperature, light, air, soil condition, soil

reaction and the antagonistic and associative effect of the

microorganism themselves. However, soil acidity (pH 4.5-5.5) is

favorable to the development of fungi; soil bacteria prefer neutral

conditions (pH 6.5-7.5), and actinomycetes thrive well in slightly

alkaline condition.

g<E3^<RM imWDVCTKm 13

Microorganisms play an important role in biochemical

transformations, such as organic matter decomposition, humification,

nitrogen fixation, ammonification, nitrification, oxidation and reduction

(Bollag and Henninger, 1976; Gowda et al., 1976; Vlassak, 1975; Gaur

and Misra, 1977;). Thus, they help the soil development and tend to

maintain the soil fertility. (Russel, 1973). Soil microorganisms are

classified as (i) microflora and (ii) microfauna.

(i) Microflora:

It comprises of the following:

Bacteria:

Bacteria, single-cell organisms, are one of the simplest and

smallest forms of life known. Soil bacteria are either autotrophic or

heterotrophic. Autotrophic bacteria have the ability to oxidize (or

reduce) the selected chemical elements in soil. Thus, through

nitrification, selected bacteria oxidize ammonium compounds to the

nitrate form, and others complete the oxidation to the nitrate form.

Other bacteria are responsible for oxidation and/or reduction of sulphur,

iron, manganese and carbon monoxide, etc.

Fungi:

Fungi are plants without seed, stem, leaf and root and have no

chlorophyll. Their biomass varies with the kind of soil. They take part in

gEm.^^flL imstimocTicm 14

a number of processes, of which their role in the decomposition of

complex organic substances is the most important. They also play a

dominant role in the synthesis of lignoprotein complex and in the

formation of other new compounds, such as polysaccharides, gums,

chitin, etc. They also decompose proteins and convert them into amino

acids and ammonia. The ammonifying capability of soil fungi, is greater

than that of many bacteria.

Actinomycetes:

Actinomycetes are filamentous and branched. They are similar to

bacteria, in that they are unicellular and have about the same diameter.

In soils, they are involved in the decomposition of all sorts of organic

substances. They have the capability of breaking down cellulose.

Proteins and amino acids are broken down and ultimately converted

into ammonia.

Algae:

Most algae are chlorophyll-bearing organisms. These are capable

of performing photosynthesis like higher plants. Soil algae are divided

into four general groups: blue-green, green, yellow green, and diatoms.

Blue-green algae are commonly available in large numbers in rice soils,

and when such land is flooded, appreciable amounts of atmospheric

nitrogen are fixed or changed to a combined form by these organisms.

g'ETm^L iwriiptDVcrioyr 15

Blue-green algae, growing within leaves of the aquatic fern Azolla, are

able to fix nitrogen in significant quantities for rice production.

(ii) Microfauna:

Microfauna comprises of the following (Sehgal, 1991):

Nematodes:

Nematodes are microscopic, unsegmented and thread like

worms. The most numerous and varied forms of nematodes are those

that live on decaying organic matter (saprophytes). But, some

nematodes, especially those of the genus Heterodera, can infest the

roots of practically all plant species.

Protozoa:

Protozoa are probably the simplest form of animal life and are the

most varied and numerous of all the micro animals in soils. Protozoa

include amoebae, ciliates and flagellates. The various kinds of chemical

reactions in soil such as oxidation-reduction (Charles and Paul, 1958),

solubilization and fixation of plant nutrients in soils (Singhal et al.,

1975), denitrification and nitrification of soil nitrogen (Noggle and

George, 1977) are all mostly controlled as a result of chemical and

microbial activities in soil (Singhal et al., 1976).

giiimvu. im>RPDi)cncm i6

The microbial activities are largely responsible for the

maintenance of soil fertility (Martin, 1973) as well as in governing soil

properties (Katznelson and Bose, 1959).

PHYSICO-CHEMICAL PROPERTIES OF SOIL

The physico-chemical properties of soil may be discussed as (i)

mechanical composition (ii) pH (iii) electrical conductivity (iv)

exchangeable cations and (v) cation exchange capacity.

(1) Mechanical composition:

The first information required about a soil is its mechanical

composition. Mechanical composition or texture means the particles of

various sizes such as gravel (> 2mm), sand (0.02-2mm), silt (0.002-

0.02) and clay (<0.002mm) fractions. The particle size distribution in

soils influences its chemical, physical and biological properties. Clay

particles, which have a large surface area, determine most of its

chemical properties. Particle size distribution in soils also influences its

water holding capacity and water supply to plants. It influences the

strength and compressibility of soils. The success of any mechanical

analysis depends firstly upon the preparation of the sample to ensure

complete dispersion of all aggregates into their individual primary

particles without breaking up the particles themselves, and secondly

upon the accurate fractionation of the sample into various separates.

gES^^iM im^RP^'UCTICm 17

(ii) pH:

Soil pH is one of tiie most important soil property, which is a

measure of the acidity or alkalinity of the soil. It gives an idea about

acedic, basic or neutral character of a soil and is defined as -log [H*].

Soil pH affects the availability of plant nutrients. Macronutrients tend to

be less available in soil with low pH. However, if the soil pH is high the

micronutrients tend to be less available due to their precipitation. In pH

range 6.0 to 6.5, nutrients are more readily available to plants. Soil

microbial population also increases in this range of soil pH.

Various soil particles, especially the clays, are saturated with

different cations. The nature and extent of cations varies from soil to

soil having different clay minerals and is reflected from their cation

exchange properties. The soil pH is also governed by the nature of

soluble salt cations saturating the soil particles over their exchangeable

sites. For instance, soils containing aluminium as its dominant cations

mostly be acidic, ranging from 3.8-5.8 in pH. On the other hand, soils

containing a lot of sodium ions will tend to be basic ranging from 8.5-

10.5 in pH.

(iii) Electrical conductivity:

It gives an idea about the total soluble salt content in soil. Soil

salinity can be measured by extent of soluble salt especially the NaCI

gEm:'SM iTmpJi'Vcnoo^ 18

and Na2S04. For saline soil, the salt content is more than 0.3% or E.G.

> 4.0 mmhos cm"\ EC of a solution increases approximately 2% per

degree centigrade. E.C of the irrigation water may vary from 0.1-0.75

mmhos cm" or below. High salinity hazard is increased in case of

irrigation water having conductance much above this range.

(iv) Exchangeable cations:

The exchangeable cations, normally present in agricultural soils,

2+ 2+ + + + +

are Ca , Mg , K , Na , NH4 and H ions. During the processes of

weathering and clay formation, the water in the immediate vicinity of the

decomposing primary mineral is rich in Ca^*, Mg^*, K* and Na* ions.

The newly formed clay mineral reacts with these ions and adsorbs them

on its surface. Ordinarily, calcium ions are adsorbed to a much greater

extent than the other ions giving rise to calcium clay. The source of H""

ion on the clay surface may be due to the decomposition of carbonic

and other acids. In addition to those, the clay minerals also found to

adsorb other cations such as iron, aluminium, manganese, zinc, copper

and other minor elements though in smaller quantity.

Cation adsorption is a function of the clay mineral; the amount of

total exchangeable cation varying with the amount of colloidal content

or clay present in soil. Hence, clay and clayey soils contain a greater

amount of total exchangeable cations than sands and sandy soils.

gT:m.<sM im'xp^'Ociicm 19

(v) Cation exchange capacity:

The cation exchange capacity is related to the fundamental

properties of soil, that represents the total quantity of negative charge,

available to attract positively charged cations in solution. It is one of the

most important chemical properties of soil, and strongly influences in

preserving the nutrients availability. The CEC of soil is expressed in

terms of milli equivalent of negative charge per 100 gm of soil (meq 100

g'"*) or in terms of centimole of negative charge per kg of soil (Cmol(p+)

kg"""). The CEC also represents the total meq 100 g" or Cmol (p+) kg"""

of cations held on the negative sites.

The CEC of soil varies with the nature and extent of clay and

organic matter contents. In case of soil, which contains very little

organic matter, the CEC may be taken to vary with the clay content of a

soil. Soils rich in montmorillonite clay possess high CEC, while those

having kaolinite as the predominant clay mineral, have low exchange

capacity.

SOIL ADSORPTION

Soil adsorption may be defined as the excess solute

concentration at the liquid soil interface over the concentration in the

bulk of the solution regardless of the nature of the interface region or of

the interaction between the solute and the solid surfaces (Mingelgrin

g'E:m<RM i^mpDVcTicw 20

and GerstI, 1983). The rate and extent to which a substance distributes

itself between the solid and solution phase over a soil are determined

by several physical and chemical properties of both the adsorbing

substance and the soil as well as by environmental conditions (Bailey

and White, 1964; Greenland, 1965; Bailey et a/., 1968) prevailing on

them.

Colloidal particles of soil possess the power of adsorbing gases,

liquids and even solids from their suspensions. The phenomenon of

adsorption is confined to the surface of the colloidal particles. The

dynamic nature of the colloidal particles of the soil is manifested

prominently in clay and humus. Because of their extremely small

particle size, they have a large surface area per unit weight. They also

exhibit surface charges that attract polar molecules and charged ions

as well as water (Weber and Weed, 1968; Bisada and Johns, 1969;

Hower, 1970; Singhal and Singh, 1976). Such attraction is of particular

significance for negatively charged colloids. Ions found in soil solutions

are in the process of ion exchange between the soil colloids and can be

replaced in equivalent amounts by other ions. For example, a hydrogen

ion released into the soil solution by plant root can exchange readily

with the potassium ion adsorbed on the colloidal surface. The

potassium ion is then available in the soil solution for uptake by the

roots of the growing crop plants (Brady, 1995).

gEimvM- im<Rp<DVciicw 21

The property of adsorption plays an important role in maintaining

the soil fertility. It is due to this property that a soil is able to hold water

and plant nutrients; keeps them available for plant use. It enables the

soil colloids to function as a reservoir of nutrients for plant use. Soluble

substances added to the soil in the form of fertilizers are retained in it

because of the power of adsorption that soil colloids possess. Thus,

properties of adsorption or ion exchange are the main criteria for

assessing the fertility status of the soil.

SOILS OF INDIA

India, constituting a part of the "Asian Continent", lies to the

north of equator. It has a geographical area of 329 m ha and lies

between latitude of 08° 04' and 37° 06'N and longitudes of 68° 07' and

97° 25'E (Sehgal, 1996). The nature and properties of its soil vary

greatly depending upon the variation in soil-forming factors and

processes. Due to the variations in different soil parameters, India has

a large variety of soils. The variety is so diverse that, India represents

all the major soil groups of the world.

Agriculture in India is highly dependent on bioclimatic conditions,

which in combination with soils, physiography and other factors,

decides the agro-ecological zones. Agro-ecological zoning is, therefore,

important for macro-scale planning, particularly in area where rainfed-

farming is practiced, which accounts for nearly three-fourth of cultivated

gEom^uiL i^fmyDVcncm 22

area in India. Based on tine relevant parameters, the National Bureau of

Soil Survey and Land Use Planning (Sehgal et al., 1992) has identified

20 agro-ecological regions, which have been found to be useful for

macro-scale planning.

The credit of the earliest classification of Indian soils goes to

Schokalskaya (1932). Thereafter, Govindarajan (1965) divided the soils

of India into 24 major soil groups. Rayachaudhuri and Govindarajan

(1971) revised the soil map of India on the basis of the US

Comprehensive System of Soil classification. The National Bureau of

Soil Survey and Land Use Planning (1985) Nagpur, India prepared a

soil map of India using the units of soil Taxonomy.

The Soil groups of India give the salient characteristics, potential,

constraints and equivalents in the US soil Taxonomy (Soil Survey Staff,

1975) and FAO legend (1991). The NBSS and LUP has been engaged

in preparing a comprehensive soil resource inventory of different states

(Sehgal, 1990) by using a latest technology to highlight the areas of

political problems (Sehgal, 1995).

On the basis of the efforts made by the National Bureau of Soil

Survey and Land Use Planning, the soil of India is classified into

following major genetic groups:

Alluvial soils:

Alluvial soils represent the most fertile land and occupy an

gEmi^iJiL imtRptDVCUCW 23

estimated area of about 75 m ha in tlie Indo-Gangetic plain and

Brahmputra Valley. These soil are widely distributed in northern, north

western and NE parts of India, including Punjab, Haryana, UP, Delhi,

MP, Assam, Bihar, WB, and coastal regions of Orissa, Gujrat, TN,

Kerala, etc. They are inherently rich in plant nutrients. These soils are

generally alkaline in reaction, but may become acidic as in NE regions

where the rainfall (> 2000 mm per annum) generally exceeds potential

evapotrasplration. These soils are further classified into Khadar (young

alluvial) and Bhangar (old alluvial). According to US soil Taxonomy

(Soil Survey Staff, 1994), such soils may be divided into various order

such as entisoils, inseptisols, alfisols and aridisols.

Black soils:

Black soils occupy an area of about 72 m ha (Sehgal and

Bhattacharjee, 1988). These soils are observed in the central, western

and southern part of India, including Maharashtra, western parts of MP,

Gujrat and some parts of AP, Karnatka and TN. These soils are highly

argillaceous, varying from 30 to 80 percent clay content. They have

high cation exchange capacity (33-55 Cmol (P+) per kg). These soils, in

the Genetic System, were classified in the order: intrazonal (grumusols)

and azonal (regosols and alluvial soils). According to soil Taxonomy,

the medium and deep black soils are grouped in the order of vertisols.

Red soils:

The red soils are widely present in the southern part of Peninsula

gEim^iM-i^NnxcxDVcTicm 24

comprising the states of TN, Karnatl<a, Maiiaraslitra, AP, MP, Goa and

tlie Union Territory of Pondiclierry, Dadra Nagar Haveli, Daman and

Diu. In the north and NE regions, these soils are occasionally observed

in parts of Bihar, WB, Assam, and UP. The red and lateritic soils,

according to the recent survey, occupy 70 m ha (Sehgal et al., 1993).

These soils are generally deficient in nitrogen and phosphorous; but are

well supplied in potassium. According to the US soil Taxonomy (1975

and 1994), they may be classified into the following orders: alfisols,

ultisols, entisols and inseptisols.

Laterites and lateritic soils:

The well developed laterites are observed on hill-tops and

Plateau of Orissa, Maharashtra, Kerala, TN and NE regions; such soils

are locally observed in AP, Karnatka and Assam. The lateritic soils

occupy 25 m ha of the total geographical area. Kaolinite is the dominant

clay mineral in these soils. The laterite and lateritic soils have been

classified, according to US soil Taxonomy, in the orders of: oxisols and

utisols.

Saline and alkali soils or salt affected soils:

In India, these soils are mainly confined to the arid and semi-arid

and subhumid regions. The salts deposited are of sodium, carbonate,

bicarbonate, sulphate and chloride with little calcium and magnesium

gEm.<RM. iTfrnyDVcncm 25

contents. These soils are observed in UP, Haryana, Rajastlian,

Maharashtra and Karnatka. In the new US Soil Taxonomy, these soils

have been classified in the following orders: aridisols, inceptisols,

alfisols, entisols and vertisols.

Desert or arid soils:

These soils are fine sandy to loamy fine sand in texture. They are

generally poor in plant nutrients with low water holding capacity. These

soils are found in western Punjab, southern Haryana and south west

Punjab. On the basis of soil Taxonomy they are grouped into orders of:

aridisols and entisols.

Forest and hill soils:

The total forest area in India is estimated as 75 m ha, which

constitutes a major geographical area. The major forest areas are

covered by tropical deciduous coniferous and tropical evergreen forests

and occur in the states of HP, J&K, UP, Arunachal Pradesh, Nagaland,

Assam, Meghalaya, Mizoram, Manipur, Orissa, MP, Maharashtra,

Kerala and Andman and Nicobar Island. These soils have been formed

under acidic conditions in the presence of acid humus and low base

status or under slightly acidic or neutral and high base status

conditions. Some other soil formation in the forest areas, developed on

sandstone, limestone or colluvium under slightly acidic to neutral

gEom^RM i^mp^vcncxH 26

condition and In sub-iiumid environment. Tliese soils are neutral to

acidic (pH ranging from 4.5 to 7.0). Some soils, developed on

calcareous and stones, show pH up to 8.0. The organic matter is varied

from 1 to 3.5%. These soils may be divided into following orders:

inseptisols, alfisols, mollisols, ultisols and entisols.

Peaty and marshy soils:

These soils occupy a limited area in localized pockets of Kerala

and NE region, and are dark to almost black in colour with abundant

organic matter contents (20-40%). They are very strongly acidic (pH as

low as 3.5 to 4,0). This may be due to the decomposition of organic

matter. As per soil Taxonomy these soils are classified into following

orders: histisols, inseptisols and entisols.

GENERAL DISCRIPTION OF ALIGARH SOILS

Aligarh covers an important area among the districts of Uttar

Pradesh. The Aligarh district lies towards north of the Ganga-Yamuna

doab within the parallels 27°29' and 28°11' north latitude and 77°29' and

78°38' east latitude. Aligarh district acquires an area of 5000 sq. km.

(including newly created Hathras district). Its alluvial deposits have a

gentle slope from north-west to south-east. There are several

depressions apart from those formed by the river valleys and drainage

lines. Topographically the district presents a trough like appearance

gEom<RM imtRp^vcucm 27

with high Ganga and Yamuna river banks at both extreme rims. It has a

semi-desert type of climate. Soils of the Aligarh district are alluvial with

little leaching and considerable accumulation of salts on the surface

(Agrwal and Mehrotra, 1951). The alluvial beds varying from reddish

brown to ash grey in colour passing through successive layers of

sands, sandy silt and clay with occasional compact beds of kankar of

an indurate character. In some places, pisolitic small concentration of

hydrated iron and manganese oxides is disseminated in the soil beds.

Salt problem in UP was recognized as early as 1876 and a

commission was set to investigate the deterioration of soils in Aligarh

district where a vast area of over 3 million acres (about 15% of the

total) consists of barren lands. Leather (1897) showed that the injurious

salts, which were the decomposition products of igneous rock under

natural weathering process and got deposited within the soil profiles.

He called such land as 'Usar'. The soil factor in the district, such as

nearness of water table and/or impedance of drainage due to the hard

kankar pan, caused intensification in the area of 'Usar' patches. The

'Usar' reclamation committee of UP (1938) reported that the alkaline

conditions in the soil of the district were mainly due to the presence of

carbonate and bicarbonate of sodium. The alkaline layers extended on

an average to a depth of 3-4 ft. deep. The distribution of saline soil,

though comparatively less also covered a considerable area of the

district. These soils were characterized by the presence of sodium

gt^mifjiL im^iptDVcncw 28

chloride and sodium sulplnate with an open texture and existed in a

state of flocculation. The alkali soils could be classed as 'solonetz' and

the saline soils as 'solon-chaks'.

According to the order of geneses of the principal soil types, the

district of Aligarh has been grouped into six natural soil regions as

indicated on the attached soil map of the district (vide fig. 5). The chief

characteristics of these soils, as investigated till the time of undertaking

the present work, are briefly indicated in the table I, II and III.

COLLECTION OF SOIL SAMPLES

According to an axiom; analysis can be no better than the

sample. This is all the more true in the collection of soil samples. The

general problem of soil sampling has been summarized by the

Association of Agricultural Chemists as follows "In view of the variability

of soils, it seems impossible to devise an entirely satisfactory method

for sampling". During the collection of soil sample hereunder described,

the importance of taking representative composite sample was kept in

mind and variations in color, texture, slope and cropping pattern were

all adequately considered. The grass and other matters were removed

from the surface soil. The soil samples (0-30 cm depth) from atleast ten

well described spots in fields were collected and then mixed well. The

different samples were collected from the following mentioned area:

cjLyL'K'ii. i:\i^>ovciio:s'

7773 t- ...

c^^ • • *

g<E3imM. imtRP^vciicw I 30 I

1. Aligarh Type I (Ganga Khadar):

This soil is distributed in narrow belt of recent alluvial tract of the

Ganga in the northeastern corner of the district in tehsil Atrauli. The

sample was collected from the field in the neighborhood of Sankra bus-

stand about two furlong from the river bank.

2. Aligarh Type II (Eastern Uplands):

This soil covers almost the entire tehsil of Autrouli except for a

thin strip of Ganga Khadar. The sample was collected from the field of

Malhepur village, situated on the Aligarh-Ramghat Road.

3. Aligarh Type III (Central Lowlands):

This soil occurs widely in the central low lying tracts of the district

in tehsil Koil and tehsil Sikandra Rao, and is underlain by the thick pan

of kankar; at some places form a stiff imperviable rock in the bottom

layers. The sample was collected from the barren field around the

Aligarh Muslim University.

4. Aligarh Type IV (Western Uplands):

The soil is mainly distributed in the tehsil of Khair, Iglas and

Hathras and forms numerous sandy ridges. The sample was collected

from Talaspur village.

QfEm.'KM' I^NnSiOOlJCTlcm 31

Table - 1

Some general characteristics of Type I (Ganga Khadar ) and

Type II (Eastern Uplands) soils of Aligarh district

Parameters

Profile development

Colour

Concretion

Texture

Lime

Soluble salts

Salt efflorescence

PH

Clay

Drainage

Water table

Main crops

Type 1

(Ganga Khadar)

Immature

Ash gray

No

Sandy to silty loam

High (slightly

calcareous)

High

Present

8.5

Very low

Imperfect

About 8 ft.

Millet, Wheat, and

Barley

Type II

(Eastern Uplands)

Mature

Light brown

No

Sandy-loam, friable

Low (non-calcareous)

Very low

Nil

7.2

Moderate

Good

About 22 ft.

Oat, Maize, Black

gram. Barley, Wheat

and Pea

g'ESm^RJlC IjmtpDVCTKXK 32

Table - II

Some general characteristics of Type III (Central Lowland)

and Type IV (West Uplands) soils of Aligarh district

Parameters

Profile development

Colour

Concretion

Texture

Lime

Soluble salts

Salt efflorescence

PH

Clay

Drainage

Water table

Main crops

Type III

(Central Lowlands)

Mature

Whitish gray

Thick kankar pan

Clayey loam, compact

High (calcareous)

High

Present

9.2

High

Very poor

About 16 ft.

Sugar cane, Barley and

Paddy

Type IV

(West Uplands)

Mature

Reddish brown

No

Sandy loam

Low (faintly alkaline)

Very low

Nil

7.9

Low

Excessive

About 25 ft.

Oat, Maize, Barley,

Wheat and Pea

gE:m<RjiL lOfmpinJcTicm 33

Table -

Some general characteristics of Type V (Yamuna Khadar) and

Type VI (Trans Yamuna Khadar) soils of Aligarh district

Parameters

Profile development

Colour

Concretion

Texture

Lime

Soluble salts

Salt efflorescence

PH

Clay

Drainage

Water table

Main crops

Type V

(Yamuna Khadar)

Not fully mature

Dark gray

Kankar at bottom

Clay

Rich (moderately

calcareous)

very high

Present

8.0

High

Poor

About 5 ft.

Wheat and Gram

Type VI

(Trans Yamuna Khadar)

Mature

Brownish gray

Kankar at lower depth

Loam

Average (non-calcareous)

Average

Not prominent

7.5

Moderate

Restricted

About 12 ft.

Pea, Barley and Maize

g<E3FE^(jiL im^pDVcncm 34

5. Aligarh Type V (Yamuna Khadar):

The overflow of Yamuna in the extreme of north-west of the

district has given rise to the formation of this type of soil in Khair tehsil.

The soil is hard with cloddy structure and gets puddled up when wet.

The sample was collected from Tappal Town, situated on the bank of

Yamuna river.

6. Aligarh Type VI (Trans Yamuna Khadar):

The tract of land containing this soil lies parallel to the Yamuna

Khadar in the form of narrow belt about six miles wide. The soil is

associated with kankar and is slightly stiff. The sample was collected

from Jattari Town in Khair tehsil.

PLANT NUTRIENTS

The seventeen chemical elements, that are important for proper

growth of plants, are called plant nutrients. Deficiency of these

elements causes several diseases and affects the life processes of

plants (Arnon, 1950). The physiological function of these elements

depends on their concentration. Too much of some plant nutrients, may

cause a nutrient imbalance in plants and adversely effects the

population of microflora and microfauna (Kuster and Grun, 1984). The

essential elements are divided into two main groups: non-mineral and

mineral nutrients.

g<EM:<RJlL imiRPOVCTlCXN' 35

Non-mineral nutrients:

The non-mineral nutrients are Jiydrogen, oxygen and carbon.

They are found in the form of air and water. Plants use the energy from

the sun to change carbon dioxide and water into starch and sugars.

Mineral nutrients:

Out of the essential elements, thirteen mineral nutrients, which

usually come from the soil, are dissolved in water and absorbed

through the plant's roots. The mineral nutrients are further divided into

two groups (i) macronutrients and (ii) micronutrients.

(i) Macronutrients:

Macronutrients are divided into two more groups (a) primary and

(b) secondary nutrients.

(a) Primary nutrients:

The primary nutrients are:

Nitrogen:

Nitrogen is a critical component of enzymes, amino acids and

proteins, which control the metabolic processes, required for plant

growth. It is also an integral part of the chlorophyll molecule and thus,

plays a key role in the photosynthesis. An adequate supply of nitrogen

g<Em:<Rjic im^iptDVcTiaN' 36

is associated with vigorous growth and dark green colour of the plant.

Nitrogen deficiency is characterized by reduced plant growth and a pale

green or yellow colouring in leaves (Chapman, 1968). If the deficiency

is severe, the affected area of the leaf eventually turns brown and dies.

Phosphorus:

Phosphorus is a critical component of nucleic acids. So, it plays

an important role in plant reproduction, of which grain production is an

important consideration. This mineral is often found in large quantities

in seed and fruit. Phosphorus is essential in the biological energy

transfer process (Mengel, and Kirkby, 1982), which is of vital

significance for the life and growth of plants (Khasawneh, 1980).

Adequate phosphorus is characterized by improved crop quality,

greater straw strength, increased root growth, and earlier crop maturity.

Phosphorous deficiency is indicated by reduced plant growth, delayed

maturity and smaller fruit size. These symptoms may be revealed

through purple colouring, particularly in young plants.

Potassium:

Potassium in plants, stay in a mobile form rather than as an

integral part of any fixed components. It helps to maintain self-

permeability, aids in the translocation of carbohydrate (Webster and

Varner, 1954), keeps iron more mobile in the plant and increases the

resistance to certain diseases. It also helps in the building of protein,

g'E^mvM' imtRpovciicxN' 37

photosynthesis, fruit quality and reduction of diseases. High potassium

in solution also affects the osnnotic pressure of the soil solution and

makes the water less available for plant use. Deficiency of potassium is

reflected in plants by white spots on the leaf edges, chlorosis and

necrosis. Potassium deficiency can also occur in young leaves at the

top of high yielding and fast maturing crops.

(b) Secondary nutrients:

The secondary nutrients are:

Calcium:

Calcium is an integral part of plant cell walls. It provides for

normal transport and retention of other elements as well as strength in

the plant. Sources of calcium are dolomitic lime, gypsum and super

phosphate. Calcium deficiency causes reduction in crop growth but, are

rare in agricultural soils.

Magnesium:

Magnesium is a key component of chlorophyll, essential for

photosynthesis. It also activates many plant enzymes, needed for

growth of plants. Magnesium is best supplied by limestone that contains

this nutrient. White strips between the leaf veins characterize the

magnesium deficiency.

gm>EvuL imtRpDVcncw 38

Sulphur:

Sulphur is a common component of proteins and vitamins, wliicli

are essential for plant and animal growth (Tabatabai, 1986). It also

helps in chlorophyll formation. Sulphur improves root growth and seed

production. Rainfall supplies significant amounts of sulfur. Sulfur also

occurs in volatile compounds, responsible for the characteristic taste

and smell of plants in the mustard and onion families. Sulfur deficient

plants have a general yellowing and are very spindly.

(ii) Micronutrients:

Micronutrients are those elements, which are needed in only very

small quantities for plant growth (Nicholas and Egan, 1975). There is

considerable variation in the specific role of the various micronutrients

(Mengel and Kirkby, 1982) in plants and microbial growth processes.

The micronutrients are:

Boron:

Boron helps in the utilization and regulation of other nutrients.

Boron is essential for seed and fruit development. Sources of boron are

organic matters and borax. A boron deficiency decreases the rate of

water absorption, root growth and translocation of sugars in plants.

Copper:

Copper is important for reproductive growth and is involved in

g'ESm^UiL HmRpcDVCTKW 39

photosynthesis and respiration (Arnon, 1949). It also stimulates

lignification of plant cell walls and helps in the utilization of proteins.

Deficiency of copper has been reported in numerous plants, although

they are more prevalent among crops growing in peat and muck soils.

In corn, copper deficiency causes yellowing and stunted to the

youngest leaves.

Iron:

Iron is structural component of porphyrin molecules such as,

cytochromes, hemes, hemating, ferrichrome and leghemoglobin. These

substances are involved in the oxidation-reduction reactions in

respiration and photosynthesis. It involves in chlorophyll formation and

degradation and also in the synthesis of proteins, contained in the

chloroplasts. Source of iron are the soil, iron sulphate and iron chelate.

A deficiency of iron is indicated in the young leaves of plants. Iron

deficiency also reduces the chlorophyll production, which results in the

characteristic chlorosis symptoms of iron stress. An excess amount of

iron is hazardous to plants.

Manganese:

Manganese is considered to be essential for photosynthesis,

respiration and nitrogen metabolism (Stiles, 1961). It also takes part in

oxidation-reduction process and in decarboxylation and hydrolysis

reactions. Soil is a source of manganese. Its deficiency causes gray

speck of oats, marsh spots of peas and speckled yellow of sugar beets.

gEymtRM nmstiyDVcticm 40

Wheat plants grown in Jew manganese, are often more susceptible to

root rot diseases. Excessive amounts of manganese are also harmful to

plants.

Molybdenum:

Molybdenum is a component of the enzyme nitrogenase, which is

essential for the process of nitrogen fixation. It is also present in the

enzyme nitrate reductase, which is responsible for the reduction of

nitrates in soil and plants. Small grains and some row crops are more

tolerant to low levels of available molybdenum.

Zinc:

Zinc plays a vital role in protein synthesis, in the formation of

some growth regulating hormones like indole acetic acid and in the

reproductive process of certain plants. Reduced growth hormone

production is common in zinc deficient plants and causes the

shortening of internodes and smaller than normal leaves.

Chloride:

Chloride is known to influence photosynthesis and root growth. A

useful function of chloride is as the counterian during rapid potassium

fluxes, thus contributing to turgor of leaves and other plant parts.

Chlorosis in younger leaves and overall wilting of the plants are the two

most common symptoms of chloride deficiency.

q^^m<SiM' ismtsyDVcTioTf 41

Cobalt:

Cobalt is essential for the symbiotic fixation of nitrogen, in

addition, legumes and some other plants have a cobalt requirement,

independent of nitrogen fixation, although the amount required is small

as compared to that for the nitrogen-fixation process. Cobalt is also

essential for the growth of symbiotic microorganisms, which are

responsible in the formation of vitamin B12. Improved growth,

transpiration and photosynthesis with application of cobalt have been

observed in many plants.

SOIL THIN-LAYER CHROMATOGRAPHY

Thin-layer chromatography (TLC) is considered as powerful tools

for the separation and identification of organic and inorganic

compounds from their complex mixture. It gives a satisfactory result,

where other methods fail. The TLC is simple, rapid, inexpensive,

sensitive and selective and it requires very little working materials. The

technique of TLC was first introduced actively for the separation of fatty

esters by Kirchner et a/.(1951) and now it has become one of the most

important technique employed in virtually every branch of chemistry for

the separation, identification and isolation of pure compounds. Some

useful and comprehensive text on the application of TLC for organic

compounds has been compiled by Stahl (1966). A variety of literature is

also available describing the TLC of lipid molecules (Bobbitt et al.,

gEmm^ iifi^&^'VctKm 42

1968; Kirchner, 1967; Zweig and Sherma, 1972; Touchstone, 1973;

Issaq and Barr, 1977).

Helling and Turner (1968) first introduced the soil TLC for the

determination of pesticides movement using different types of soils as

static phase. Rhodes ei al. (1970) used soil TLC for the determination

of agrochemicals in soil. Chapman et al. (1970) also used soil TLC for

measuring the relative mobility of herbicides. The work was further

compiled by Helling (1971) in three reports dealing with pesticides

mobility in soils (1) parameters of TLC (2) application of soil TLC and

(3) influences of soil properties.

Soil TLC method has been successfully used by Singhal and

Singh (1977) for the mobility of trace metals, Singhal et al. (1978) for

the mobility of amino acids. Khan et al. (1982) for the mobility of heavy

metals. Khan et al. (1985) for the effect of organic acids and bases on

the heavy metals mobility. Khan and Khan (1986) for the mobility of

organophosphorous insecticides in soils as affected by soil parameters

and Khan et al. (1999) for the mobility of amino acids in soil as affected

by some pesticides. Soil TLC technique has now become an important

analytical tool in describing the mechanism involved in the mobilization

process of organic and inorganic compounds through soils.

Soil TLC is an adsorption chromatography, where the adsorbent

is a thin-layer of soil deposited on a glass plate. The soil components

gEm-vu:. imspfDVcno^r 43

provide an adsorptive or static pliase, where adsorption and desorption

takes place very rapidly and reversibly. Thin layer, prepared by

spreading the slurry of soil as uniform film (0.15-2.0 mm) with the help

of an applicator, is allowing to stand overnight for drying (Bremner et

al., 1961; Bremner et al., 1962; Michalec et al., 1962). Better results

are obtained if the layer is thin, because the spray reagent is much

more sensitive when it does not have to search for tiny amounts of

substance in a large amount of adsorbent. On the contrary, when the

work is preparative in nature, it is advisable to use a thick layer (Cerny

et al., 1961) so that the maximum amount of material can be separated

in a single chromatogram. The amount of substances applied depends

upon thickness of the layer and visualization procedure, which are

inversely proportional to each other and also on the nature of

adsorbent. The compound under investigation is spotted at fixed

scribed place (about 4 cm above from the bottom) on the plate with the

help of micropipette. The mobile phase level in the closed glass jar

always remains about one cm below the spots.

Chromatograms are then developed in a closed chamber using

selected developer. After the solvent ascends up to scribed line (up to

10 cm from the base line of the plate), the plate is removed, dried and

detected by spraying a suitable chemical as detector. The mobility in

terms of Rf value can be calculated by the formula.

R value- *^'^^^"^^ moved by solute ^ Distance moved by developer

gEm.'HM' iTfT^tsyDVcticm 44

AMINO ACIDS

There are twenty naturally occurring amino acids that build

animal proteins. Amino acids constitute a very important group of

organic compounds conforming the general formula, R-CH(NH2)C00H

(except proline). Amino acids in solution at isoelectric pH are mainly

dipolar ions, called zwitter ions; the molecules are left with no net

charge. These are classified as: acidic, basic, neutral-polar and neutral

non-polar. Because the alpha carbon of an amino acid is an asymmetric

carbon, each amino acids can exist in the form of two enantiomers.

These enantiomers are called L-isomers and D-isomers. Amino acids,

present in living systems, are almost exclusively L-isomers except

some D-isomers (Bege et al., 1993).

With some exception, bacteria and plants can synthesize all of

amino acids the need from simpler substances. If the proper raw

materials are available, the cells of humans and animals can

manufacture some, but not all, of the biologically significant amino

acids (Russel, 1996). Those, that humans and animals cannot

synthesize, must be obtained through their diet, known as essential

amino acids.

The nitrogenous organic compounds in the soil are derived from

plant and animal residues, root exudats and microbial cells (Verhoeven

et al., 1956; Hale and Moore, 1979). Among these nitrogenous organic

gT3^<RM im^CXDVClKXN' 45

compounds, the amount of the free amino acids, that can be detected in

soils, is very small (Sperber, 1958), attesting to the fact that these

compounds are rapidly immobilized or transformed. Paul and Schmidt

(1960 and 1961) showed that the total amount of free amino acid

nitrogen in soils rarely exceeds 2 )ag per gm of soil. The amounts of free

amino acid nitrogen are related to the level of microbial activity

(Putman, 1959; Paul and Sehmidt, I960;). It is likely that, some of the

combined amino acids in soil remain in the form of mucopeptides and

teicholic acids and these compounds are essential constituent of

bacterial cell wall. Some growth stimulators of plants contain amino

acids (Takahashi, and Okano, 1999)

In soils with high clay content, there is a greater tendency to

preserve potentially labile compound such as amino acids (Sorenson,

1972; Sorenson, 1975; Balesdent et al., 1988). The use of fertilizers

affects the contents of free amino acids in soils (Udaipur et al., 1998),

which favours the growth of amino acid decomposing, ammonifying and

nitrifying bacteria. Amino acid contents have been found highest in both

extremities of plants, grown under the conditions of the maximum

photosynthesis in soils with moisture content of above 36% of

saturation (Lagun, 1972). The amino acid contents of the soil

decreased at end of the growth period.

Talibuddin (1955) studied the adsorption of some amino acids by

montmorillonite at pH 2.5 and 4 and concluded that the amount of

gEjmtM- nmfRp^DVcncm 46

adsorbed amino acids was a function of basicity and size of the amino

acid molecules. Sieskind and Wey (1959) observed that the adsorption

of amino acids found a linear relationship between the amount of

adsorbed amino acids in meq 100 g" montmorillonite at pH 2. Singhal

et al., (1978) observed that the pH, particle size and types of soils

affect the mobility of amino acids. It has been observed that amino

acids mobility was affected by pesticides in soil (Khan et al., 1999).

PESTICIDES

The term pesticide include any substance intended for

preventing, destroying, attracting, repelling or controlling any pest

including unwanted species of plants and animals during production,

storage, distribution and processing of food, agricultural modifies or

animal feed (Ware 1978). These are the largest group of poisonous

substance, available in gaseous, liquid and solid forms. When applied

at recommended rates, the concentration of pesticides present in soil

does not significantly alter the microbial activity that are important to

soil fertility.

At present more than 10,000 different pesticides are widely used

to control or destroy the different types of unwanted pests and weeds.

Pesticides may be classified into following major categories:

insecticides, herbicides, fungicides, rodenticides, nematicides, acricides

etc., which are in commonly used:

g'E^m.'RM' imtRpiDVcncw \ 47 I

Insecticides:

Chemical substances, which are used for killing insects are

known as insecticides. According to their mode of action, they are

usually classified as: stomach insecticides which are lethal only to

insects that ingest them, contact or external insecticides which destroy

the insect simply by external bodily contact and fumigants which act on

the insect through respiratory system. On the basis of chemical nature

of insecticides, they may be classified as (1) inorganic (ii) natural and

(ill) organic insecticides.

(i) Inorganic Insecticides:

Before the introduction of modern insecticides, the inorganic

insecticides as well as those of plant origin were extensively utilized for

plant protection. In recent years, inorganic insecticides have been

generally replaced by organic formulations in various applications. The

major disadvantage of inorganic insecticides is in their toxicity to man

and other warm-blooded animals. Some inorganic insecticides are: lead

arsenate, calcium arsenate, paris green, sulphur and compounds of

halogen, copper, phosphorus, mercury, sulphur, etc.

(Ii) Natural Insecticides:

Plant materials yield some of the most widely used insecticides.

Nicotine, rotenone, allethrin etc., are well known examples of natural

insecticides.

g<Em:<RM im>RP^vcncw 48

(iii) Organic insecticides:

Organic insecticides include: organo-pliospliorous and chlorina

ted hydrocarbons.

Organo-phosphorus hydrocarbons:

These have generally a very short residual action. They control a

wider range of pests than chlorinated hydrocarbons. They are, however,

rarely used for the control of soil pests. These insecticides are

poisonous to mammals and should, therefore, be handled with great

caution. Parathion, malathion, dimethoate, tetraethylpyrophosphate

(TEPP), dichlorvos, phosphamidon, etc., are some common examples of

organo-phosphorus hydrocarbons.

Chlorinated hydrocarbons:

They have generally a long residual action and are generally less

toxic to mammals than phosphorous compounds. DDT was the first to

be used as a synthetic chlorinated hydrocarbon. Although, DDT was

prepared by O Zeidler in 1874 but, its insecticidal properties were

discovered by a Swiss Chemist, Paul Muller in 1939. Some common

chlorinated hydrocarbons are: benzenehexachloride (BHC), endosulfan,

methoxy-chlor, chlordane (1068), heptachlor, aldrin, dieldrin, endrin,

toxaphene, etc.

gEm.<RjiL imtRp^vcncm 49

Herbicides:

Herbicides are used to control unwanted weeds in cultivated crop

plants. The biological activity of herbicide molecules is a reflection of

their ability to penetrate into plant tissues, resist detoxication (or be

transformed into toxic products), and interfere to particular

physiological or biological processes in plants. However, many

herbicides, which are effective to monocots, are injurious to dicots. For

example, atrazine and diuron interfere the photosynthesis while

glyphostate, sulfonylureas and imidazolone block the synthesis of

essential amino acids (Mehta et at., 1993). Herbicides include (i)

triazine (ii) substituted ureas (IN) carbanilates (iv) phenoxyalkanoic

acids (v) chlorinated aliphatic acids and (vi) amitrole.

(1) Triazines:

The triazine pesticides are among the most widely used

agricultural pesticides. These are primarily used as pre-emergent

herbicides. They conform to the general formula:

. . ^ ^ N / ' ^ R ,

where,

Ri and R2 are lower alkyls and x is a chloro (-CI) or methoxy (-OCH3) or

gE;m<RM- i7mm>i>cncm 50

thio methyl (-SCH3), etc. Some of the commonly used triazine herbicides are:

simazine, cyanazine, atrazone, prometone, ametryne, prometryne, etc.

(ii) Substituted ureas:

The asymmetric phenylureas constitute one of the most important

groups of herbicides that are being used. They are non-volatile, non-

corrosive and non-flammable organic compounds. They are absorbed

by plants from the soil and translocated upto the tops through the

transpiration stream (Minshall, 1954). At low dosage, they are selective

and are used to control the seedling growth of weeds in certain crops.

At higher dosage, they are used as soil sterilants in non-agricultural

areas. Some commonly used substituted ureas are: phenyldimethyl-

ureas and chlorophenyldimethylureas.

(iii) Carbaniiates:

They are used as pre-emergence and post emergence

herbicides. These compounds are readily degradable by

microorganisms. They are lost through volatilization, if they are applied

to wet soil. Some of the most extensively used compounds of this group

are: swep, propham, chlorpropham, barban, etc.

(iv) Phenoxyalkanoic acids:

These are generally employed as pre-emergence treatment in

q^sms^iimis^vcncm 51

conjunction with other herbicides, because of their short residual life.

The menribers of this group are used in foliar treatment. Some of the

important phenoxyalkanoic acid herbicides are: 2,4-dichlorophen-

oxyacetic acid (2,4-D), 2,4,5-trichlorophenoxyacetic acid (2,4,5-T),2-

methyl-4-chlorophenoxyacetic acid (MPC), silvex, etc.

(v) Chlorinated aliphatic acids:

Various chlorinated aliphatic acids such as 2,2-dichloropropionic

acid (dalapan), trichloroacitic acid (TCA) and 2,2-dichloroisobutyric acid

(FW-450), etc. have been used as chlorinated aliphatic herbicides.

(vi) Amitrole:

It is a substituted triazole (3-amino-1,2,4-triazole) herbicide, which Is

used primarily due to its resistivity towards microorganisms (Ashton, 1961).

However, it was la.ter observed that chemical as well as microbial degradation

both may be experienced in this compound.

Some other compounds, which are used as herbicides are:N-

aliphatic carbamates, dipyridyls, etc.

Fungicides:

Fungicides have been used for the control of plant pathogens.

Fungicides, on application to various plant parts, are absorbed by the

plant tissues and translocated either upwards or downwards and control

g<E3^<RjiL imtRp^DVcncm 52

diseases away from the site of application. They may be described

under the following categories:

(i) Benzimidazoles and thiophanates:

These are used as foliar sprays, seed, soil and post-harvest dip

treatment (Delp and Klopping, 1968).

(ii) Oxathiim derivativas:

They possess marked activity against basid:omycetes i.e., stunt

and rust fungi in particular (VonSchmeling and Kulka, 1966).

(iii) Organophosphorus fungicides:

These are used against blast of rice, powdery mildews, soil-borne

diseases, etc.

(iv) Phenylamides:

The phenylamides on the whole are very effective against

oomycetous fungi.

(v) Hydroxy pyrimidines:

KoLve

The hydroxy pyrimidines^been used on cereals, cucurbits, roses

and apples, against their respective powdery meldews.

(vi) Ergosterol biosynthiesis inhibitors:

They are very selective in their mode of action and are

particularly by effective against rust, bunts and mildews of cereals.

gE:ffE<s^M- im<ftpi>vcno9f 53

Rodenticides:

These are used to control and kill rats, mice, squorrels, ground

hods, field rodents, etc. Warfarin, sodium fluoroacetate, nor-bromide,

calcium cyanide, etc., are the common chemicals that are used as

rodenticides. Warfarin acts as anticoagulant and depresses prothrombin

level, which is a blood protein required for blood clotting. Sodium

fluoroacetate, an extremely hazardous rodent killer, causes

hyperstimulation of central nervous system and affects hearts action.

Nor-bromide constricts small peripheral blood vessels of rats (Sharma

and Kaur, 1995).

Nematicides:

Nematicides are used for controlling underground practice

nematodes that attack on the root hairs of plants. Nematode problem

occurs in all areas of the world where crops are grown. It causes more

damage in "old tired soil", that is those, which have been cultivated for

a long period of time. The application of nematlcidal chemicals may

decrease crop losses (Evan, 1973; Blunt, 1975; Hummel, 1983).

Nematicides are of two types (i) fumigants and (ii) non-fumigants.

(i) Fumigants:

Soil fumigants give protection to crop plants for prolonged period,

posses a high proportion of active ingredients with effectiveness

g^-im^L iy^^cDVcrio!H I 54 I

against plants (Hollis, 1958). These are usually formulated as liquids,

which vaporize and move through air spaces in soil after vaporizing.

Some of the important fumigants are: methylbromide (MB), 1-3-

dichloropropane, methylisocyanate, etc.

(ii) Non-fumigants:

They are also called as systemic nematicides. They are often

formulated in the from of granules but, some are also available in liquid

forms for spraying on soil or foliage. Some common examples of non-

fumigant nematicides are: carbobofuran, fenamiphos, oxamyl etc.

Acricides:

These are generally employed for the control of pests like mites

and ticks, which are actually parasites living on juices sucked from fruit

and other parts of plants. Tetraethyl pyrophosphate, nemagon, lelone,

oxamyl, etc., are commonly used acricides.

Although, the use of pesticides is necessary for the modern agro-

technology but, they also affect to non-target organisms (Audns, 1970;

Ojha ef a/., 1992; Ignacimuthu and Sarvana, 1994; Murugappan and

Guna, 1996; Zhou ef a/., 1997; Xing et al., 1998). Recent reoorts have

indicated that our environment is chronically polluted by pesticides

(Heuper and Conway, 1964; Viswanathan, 1997; Rath et al., 1998; Zhu

and Hu, 1997; Xing et al., 1998), which is increasing tremendously in

g^mE^iM im^RfxDVciicw 55

various parts of the world. Therefore, there is an urgent need to control

and prevent the environmental pollution, posed by pesticides, through a

scientific approach.

SURFACTANTS

Surfactants are surface-active agents that dissolve partly in water

and partly in organic solvents. Surfactants are divisible into three main

groups: anionic, cationic and non-ionic.

Anionic surfactants are characterized by the possession of a long

non-polar portion to the molecule attached with the compact

hydrophilic, ionized group at the end. For example.

If o _

Na

where.

Alkyl benzene sulphonate (LBS)

R—S—O Na

Linear alkyl sulphonate (LAS)

R = Hydrocarbon chain 12-19 carbons.

1 { Ace. No )'")|

gEm.vi^ iTrntsyDVcncm 56

Cationic surfactants are usually quaternary ammonium salts. For

example,

R4—N—R2

where,

Ri, R2 and R3 are short hydrocarbon chain and R4 is aromatic. X

is halogen or acid group.

Non-polar surfactants are more soluble in hydrocarbons and

similar oily liquids. The most common feature of the non-ionic

surfactants is:

R-0-(CH2-CH2-0);;—H

Pre-alkyI polyoxyethylene alcohol.

CH3-{CH2)8 - ^ 7 -0—(CH2-CH2-0^ -H

Polyoxyethylenenonyl phenolate.

Surfactants lower the surface tension of the liquid in which they

are dissolved. They give stable emulsions or suspensions with soil

particles that need removal (Yeom et al., 1996). Consequently,

surfactants are used for remediation (Lee, 1998; Doong et al., 1998;

Yuan and Jafvert, 1995; Nivas et al., 1996; Huang et al., 1997) and

bioremedation (Tiehm eta!., 1995; Volkering et al., 1998; Boopathy and

gEjm^UiL imsfpcDVcncm 57

Manning, 1999) of soil pollutants. These are also used as enhancing

agents for the biodegradation of organic pollutants (Jahan et al., 1995;

Kim et al., 1998) and skin cleaners (Thau, 1997). Surfactants have

been demonstrated to be effective when applied to water repellent soils

for increasing infiltration, decreasing erosion and increasing seed

germination (Latey et al., 1962; Pelishek et al., 1962; Osborn et al.,

1964; Osborn et al., 1967).

The use of synthetic detergents containing surfactants, released

in soil and water, causes pollution problems (Valoras et al., 1976). They

have been found to change the migration, population, growth and

activity of soil microorganisms (Poremba et al., 1991; Kowalczyk, 1993;

Kozhevin et al., 1994). Surfactants are also toxic to animals (Lewis,

1991; Lewis, 1992; Roguet et al., 1992) and plants (Hall et al., 1989;

Pfleeger et al., 1990; Kloepper et al., 1996).

ORGANIC ACIDS

Organic acids are naturally occurring substances (Vandercook,

1977) which exist in soils by the transformation of soil organic

components and plants. The low molecular weight organic acids are

produced during microbial decomposition of organic materials

(McKeague et al., 1986; Fox and Comerford, 1990; Fox, 1995 ). These

acids are also formed during degradations of humus by UV light (Allard

etal., 1994; Dahlen et al., 1996).

gT3^<RJiL imtRPOVCTlOTi 58

A number of aliphatic acids such as, oxalic, citric, formic, acetic,

malic, etc., have been found in soils and litters (Stevenson, 1967;

McKeague et al., 1986; Tan, 1986). It has been suggested that they are

important in the podzolization process (Aritovskaya and Zykina, 1977;

Lundstrom et al., 1995). In addition, these compounds have a high

carboxyl contents, may therefore be important for the buffering capacity

of the soil solutions.

TRACE ELEMENTS AND THEIR EFFECTS

Out of the seventeen elements to be essential for plant growth;

eight are required in very small amounts, and are called trace elements

or micronutrients (Nicholas and Egan, 1975; Stevenson, 1986). These

are iron, manganese, zinc, copper, boron, molybdenum, cobalt and

chlorine. The dominant form of trace elements, that remained in the soil

solutions, are found mainly iron as Fe^*, Fe^*, Fe(0H)2*, Fe(OH)^*;

manganese as Mn^*; zinc as, Zn^\ Zn(OH)*; copper as Cu^\Cu(OH)*;

molybdenum as M0O4 " HMo04'; boron as, H3BO3, H2BO3'; cobalt as

Co^^ and chlorine as Cl' (Lindsay, 1972). Plants as well as

microorganisms require traces of these elements. Non-availability of

these elements in soils may result in the manifestation of specific

symptoms in plants.

There is a considerable variation in the specific roles of the

various trace elements in plants and microbial growth processes

g<E!m<RM' lofuw^vcncm 59

(Mengal and Kirkby, 1982; Marschner, 1986). Some of them are found

to participate in tlieir cellular enzymes activities and other important

processes. For example, copper, iron and molybdenum are capable of

acting as "electron carriers" in enzyme systems that bring about

oxidation-reduction reaction in plants. These trace metals are required

in such type of reactions, as are essential for plant development and

reproduction. Zinc and manganese are needed in the function of

enzymes, systems necessary for important reaction in plants

metabolism. Molybdenum is essential for certain nitrogen

transformation in microorganisms as well as in plants (Stiles, 1961).

Cobalt is essential for nitrogen fixing microorganisms. Cobalt is also

needed in nodules of legumes and alder as well as in nitrogen fixing

algae. Chloride acts as countarian during rapid K fluxes, which may

have important biochemical and /or biophysical consequences. Boron

plays an essential role in the development and growth of new cell in the

plant meristem.

The physiological function of trace elements depends on

concentration. These are essential for physiological function at low

concentration. On the other hand, they have a deleterious effect on

living organisms at higher concentrations (Dijkshoorn et al., 1979; Khan

and Khan, 1983; Brookes and McGrath, 1984; Brookes et al., 1984;

Sharaf, 1996; Anuradha and Raju, 1996; Doelman, 1998; Baath et al.,

1998), at which they may be termed as toxic. For example.

gE^fmuM- nfm^yDVcncm 60

molybdenosis, a disease in cattle is caused by an imbalance of

molybdenum and copper in their diet. It is reported in literature that Cu,

Zn, Fe, Mn, Co, Mo, Cr, Sn, Si, Ni, I, Se, F, V, and As are essential for

animal health (Underwood, 1977). Many of these, Cu, Zn, Fe, Mn, Co,

Mo, Cr, Sn, Ni, V and As are heavy metals. There is increasing

evidence that nickel may be essential for the growth and reproduction

of plants (Hutchinson, 1981; Walker et al., 1985). The chemical

behaviour of trace elements has been extensively studied in relation to

soils and plants (Bowman et al., 1981; Hodgson, 1963; Tiller et al.,

1979; Zhu and Liu, 1986).

Utilization of sewage and industrial wastes as a source of

micronutrients in most of the developing countries is very common.

Composition of these huge waste materials has been studied by various

workers (Stover et a!., 1976; Anderson and Nilsson, 1972). It was found

that harmful metals constitute a major part of its composition along with

some useful micronutrients. Metals added in soils remained in the

upper layer extended to few inches (Page and Chang, 1975; Williams et

al., 1980), because of various immobilizing mechanisms like

precipitation and specific adsorption on metal oxides, clay minerals and

organic matters. Considering the sludge and wastes as a complex

mixture of organic an inorganic species, various available forms of trace

metals exist in soil (Tills and Alloway, 1983; Gregson and Alloway,

1984).

g<E:m<RM imiRP'DVcncxN' 61

It is also Important to understand the free Ion activity of these

trace metals In soil solution and their adsorption-desorption equilibria

over soil particles. This gives information regarding the uptake and

supply of various elements by plants. Thus, there are dynamic equilibria

between various ions in solutions and the solid phase of the soil

particles (Sillen and Martell, 1971). The literature available on the role

of trace elements under Indian climatic conditions is lamentably

inadequate. More attention on the judicious utilization of trace elements

is required in the near future.

FLY ASH

Coal burning is a major source of energy production in India.

Large quantities of solid residues are generated each year (Muraka et

al., 1987) from coal burning as a byproduct containing FA (Rehage and

Holcambe, 1990). Around 60-70 million tones of FA is produced every

year from different thermal power plants in India (Saxena and Ashokan,

1998). Increasing release of FA is becoming a problem for its disposal.

The most common methods of FA disposal are land filling and settling

ponds (Suloway et al., 1983).

Fly ash contains some useful nutrients such as Ca *, Mg *, Cu *,

2+ 2- -3

Zn , SO4 , PO4 , CI etc., (Leonard and Davidson, 1959; Cope, 1962;

Hodgson and Holiday, 1966; Saxena et al., 1997) which are essential

for plant growth. Therefore, it has agronomic potential (Sikka and

giE^NmAL imtRptDVcncXN' | 62 I

Kansal, 1994) either as liming agent to neutralize soil acidity (Hodgson

et al., 1982; Masti and Keramida, 1999), boron fertilizer (Plank and

Martens, 1974; Masti and Keramida 1999) and as a physical

amendments for soils (Doyle, 1976; Chang et al., 1977) or as a soil

modifier (Saxena e al., 1996). Addition of FA to the soil also improves

the porosity of the soil and thus, increases the water retention capacity

(Campbell, 1983; Saxena et al., 1997).

Many research workers have recommended the use of FA for

increasing the crop yield of alfalfa, barley, white clover, rice, wheat and

Swiss chard (Page e^ al., 1979; Elseewi et al., 1980; Hill and Lamp

1980; Sikka and Kansal 1995; Summer et al., 1998) and for improving

the physical and chemical characteristics of the soils. Hill and Lamp

(1980) suggested that FA may be used as Mg fertilizer for pasture.

Elseewi et al. (1978) observed that plants grown on sulphur deficient

soils responded favourably to incorporation of FA into the soil. The

main contraints are still posing strains due to its high alkalinity and salt

contents (Adriano et al., 1980) and potentially harmful trace elements

like As, Cd, Pb, Sb etc. content (Kleir and Andren, 1975; Bel and

Phung, 1979; Thicke, 1988; Chandra 1997) of coal ash which are

phytotoxic and inhibit crop growth and cause deterioration to soil.

However, there Is a tremendous scope for conducting research in

relation to soil-FA amendments in the fields of agriculture.

gEim^tM J^mP'DVClKXN' 63

SOIL POLLUTION

Mankind is dependent on the soil eitlier directly or indirectly for

food, clothing and shelter. But, increasing world population and the

increasing usage of chemical doses, leads to ecological problems

because of wide dispersal of these chemicals into the soil environment.

Recent research has shown that most of the dispersed toxic, persistent

and bioaccumulative chemicals occur widely throughout the global

environment. Increasing evidence is coming forward that these

substances are having deleterious effect on biological systems (Perrin,

1998; Silant'ev et al., 1998). These substances remain in direct contact

with the soils for relatively longer periods. Thus, soil is getting heavily

polluted day by day by toxic materials and dangerous microorganisms,

which enter into the air, water and the food chain. For all this, man is

the original and basic polluting agent responsible for pollution hazards

and toxic effects. Soil pollution mainly results from the following

sources:

Industrial wastes and sludges.

Urban wastes.

Radioactive pollutants.

Agricultural practices.

Chemical and metallic pollutants.

Biological agents.

gwmi^M' imtRp(Di)ciio!K 64

Disposal of industrial wastes is the major problem responsible for

soil pollution. These industrial pollutants are mainly discharged from

pulp and paper mills, chemical industries, oil refineries, sugar factories,

tanneries, textiles, steel, distilleries, fertilizers, pesticides industries,

coal and mineral mining industries, metal processing industries, drugs,

glass, cements, petroleum and engineering industries, etc. Industrial

wastes mainly composed of organic compounds along with inorganic

complexes and non-biodegradable materials (Baldwin et al., 1983;

Jacob et al., 1994; Grosser et al., 1995; Goodin and Webber, 1995;

Bechmann and Grunewald, 1995). These pollutants affect and alter the

chemical and biological properties of soils. As a result, hazardous

chemicals can enter into human food chain from the soil or water and

may disturb the biochemical processes and finally lead to serious health

hazards.

The industrial wastes consist of calcium salts, fuel gases and

several toxic volatile elements such as arsenic, selenium, mercury, lead

and cadmium, which pose detrimental affects on environment (William

and David, 1973; Tyler, 1981; Lee, 1985).

Urban wastes comprises both commercial and domestic wastes

consisting of dried sludge of sewage. Urban domestic wastes, though

disposed off separately from the industrial wastes, can still be

dangerous. This is so because they can not be easily degraded. The

gT3^<RM. im^RpoVcucm 65

leachates from the dumping sites and disposal tanks of sewage, mixed

with industrial effluents and wastes, are extremely harmful and toxic.

Radioactive substances resulting from explosions of nuclear

devices, atmospheric fall out from nuclear dust and radioactive wastes

that penetrate into the soil create soil pollution. Radionuclides of

radium, thorium, uranium, Isotopes of potassium (K-40) and carbon

(C-14) are very common in soils, rocks, water and air. Radioactive

wastes contain several radionuclides such as strontium-90, iodine-129,

caesium-131 and isotopes of iron, which are most injurious. Recently it

has been indicated that some plants such as lichen and mushroom can

accumulate Cs-137 and other radionuclides, which get concentrated in

grazing animals.

Modern agricultural practices are also responsible for the soil

pollution to a large extent. Today, with the advancing agro-technology,

huge quantities of chemical fertilizers, pesticides and soil conditioning

agents are employed to increase the crop yield. Although, the fertilizers

are used to fortify the soil, yet they are also found to contaminate the

soil with their toxic impurities. For instance, cadmium is the main

source of low-grade rock phosphate. When the fertilizers are

contaminated with other synthetic organic pollutants, the water present

in the soil may also get polluted. Different kinds of pesticides used to

control pests and pest born diseases, are causing a stress in the

natural environment. Pesticides may get absorbed by soil colloids,

which may contaminate root crops, grown on such soils. In addition to

the fertilizers and pesticides, soil conditioners and fumigants are also

employed to the land system to increase and protect the soil fertility as

well as to kill the hazardous insects. These chemical agents are

reported to cause alteration in both agricultural and horticultural soil

areas. Animal wastes containing several pathogenic bacteria and

viruses enter into plant metabolism and ultimately reach human beings.

Synthetic chemicals and fertilizers containing trace metals

(Swaine, 1962; Tiller, 1983) are added to the soil either deliberately or

as an impurity. Metallic contaminants in soil are considered to be the

indestructible poisons and their accumulation in plants and water may

be highly dangerous.

Soil gets large quantities of human, animal and bird excreta,

which constitute the major source of land pollution by biological agents.

These biological agents are also highly responsible for huge

contamination of soils and crops by pathogens.

THE PROBLEM

The significance of soil science and agricultural chemistry are of

paramount importance in India as it is mainly an agricultural country,

where more than 80% of the population, out of 1000 millions, depends.

In order to feed^ a large population the search for a greater output in

food crop production with a low cost indigenous technology is needed

gEmi'RJiL I!f^'I^RP^'OCU(XN' 67

and therefore, different types of materials such as chemical fertilizers,

pesticides, fly ash, sewage sludge and industrial waste have been and

are being extensively tried to ascertain their feasibility the crop

production. But, excessive use of these materials has resulted in a

decline in the quality and quantity of crops due to imbalance in the

nutrient concentration and microbes in soil.

The aim of the present study was to build a ground structure and

to give a fillip to research of national and international importance in

field of soil science and Agricultural Chemistry. The thesis has been

divided into following parts:

Part I, general introduction, covers a brief account of present

and past literature in related fields of soil research.

Part II, chapter I, describes the role of pesticides such as

parathionmethyl, dimethoate, carbendazim, chloroyrifos endosulfan and

methyldemeton on the mobility of amino acids through soil TLC.

Part III, chapter II, describes the behaviour of fly ash on the mobility of

amino acids, such as lysine, glyeine, alanine and valine as examined

through soil TLC.

Part IV, chapter III, describes the effect of surfactants such as CTAB,

Triton X-100 and SDS on the mobility of trace metals viz. Pb, Cu, Ag,

Zn and Co through soil TLC.

qfEamuM- imw^vcncm 68

Part V, chapter IV, reports the effect of fly ash on the physiological

parameters such as seed germination, number of leaves, shoot length,

root length and fresh and dry shoot weight and metals uptake of barley

and wheat plants.

Part VI, chapter V, describes the effect of endosulfan pesticide on the

physiological parameters such as seed germination, number of leaves,

shoot length, root length and fresh and dry shoot weight and nutrients

uptake of fenugreek plant, which is used as spices and medicine.

gEumiiM- i9fMP<DVcnoy/ 69

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CHAPTER I

MOBILITY OF AMINO ACIDS THROUGH SOIL

AMENDED WITH SOME PESTICIDES

CUdpter I

INTRODUCTION

C j y ^ he significance of amino acids and pesticides is well known

^ - ^ In relation to soils and plants. Amino acids are either

directly added to (Rending, 1951; Sowden, 1956; Li et al.,

1999) or found to exist in soil as a result of the decomposition of proteins

and other organic matter (Wal sman and Lomanitz, 1924; Waksman and

Starkey, 1932; Scheffer et al., 1958; Summida et al., 1993; Appel et al.,

1999). The amino acids play a vital role in the biochemical processes

(Theng, 1974) and organic matter transformation in soil. Depending upon

the pH of the soil environment, amino acids can exist as cations, zwitter

ions or anions. This property enables them to be adsorbed or undergo

exchange with metallic ions on soil and clay surfaces. By virtue of their

nature, they form a variety of complexes (Talibuddin, 1955) with clays,

lignins or soil organic matter and may undergo polymerization reaction.

The products, such as indoles, act as growth promoting substances,

hormones or auxins in plant (Waksman, 1952).

CKA^T^^ I 84

Pesticides, reaching soil either by direct application or run off from

treated crop plants, get adsorbed on soil colloids, which have been

intensively studied in recent years (Kaneko et al., 1978; Yanron, 1978;

Camazano and Martin, 1987). It is found that pesticides treatment led to

both qualitative and quantitative changes in the free amino acids

composition of soils (Wainwright and Pugh, 1975; Mody and Munshi,

1990) and plants (Mehta et al., 1993). They also inhibit the synthesis of

amino acids (Kishore and Shah, 1988; Shaner, 1989; Devine et al., 1993).

It has been observed that pesticides contamination adversely effects

protein and amino acid metabolism in aquatic animals (Reddy et a!., 1991;

Patil et al., 1992). Some studies have also been done to investigate the

identification of amino acids by chromatography (Bremner, 1952;

Stevenson, 1956), mobility of amino acids (Singhal et al., 1978) and

pesticides (Khan and Khan, 1986) in soils and the effect of pesticides on

soil microbial population growth (Martin, 1964; Parr, 1974; Greeves, et al.,

1976; Alexender, 1977). But, there is a lack of literature about the effect of

pesticides on the mobility of amino acids in soil medium

In view of the significant relationship attached to amino acids, soils

and pesticides, the present study has been conducted to investigate the

mechanisms involved in the mobilization processes of certain amino acids

through pesticides amended soil.

CHAtPim. I 85

EXPERIMENTAL

For this investigation, Hirapur fine sandy loam soil (Aerie

Halaquept) sample (0-30) was collected from Aligarh Muslim University

Farm, situated in Aligarh district (U.P.). India. Physico-chemical properties

of soil were determined as follows:

Determination of mechanical composition:

The mechanical composition of soil sample was determined by

using international pipette method (Piper, 1950) as:

Apparatus and reagents required:

7, 200, 270 Mesh sieves (BSS)-

Graduated measuring cylinder.

Constant temperature water bath.

Pipette.

Petridish.

Electronic balance.

30% Hydrogen peroxide.

0.2N Hydrochloric acid.

Sodium oxalate.

Procedure:

A 10 gm of dried, crushed and sieved through 7 mesh sieve (BSS),

soil sample was dispersed in water after treating with 30% H2O2 and 0.2N

CHMT^<H I 86

HCI using 50 ml sodium oxalate (8g/liter) as dispersing agent. The

percentage of sand was calculated from the weight of the residues left

behind on 200 mesh sieve (BSS). The suspension was then diluted to 500

ml and transferred to a graduated measuring cylinder, which was

immersed in a constant temperature water bath at 25 ± 1°C throughout the

coarse of pipetting. 10 ml of the sample was pipetted out carefully at

specified intervals (4 mins. 15 sec. and 7 hrs. 5 mins.) of time from a

depth of 10 cm andjthrough 270 mesh sieve (BSS). The suspension, so

obtained, was dried in a petridish and weighed and thus, the percentage

of clay was calculated from weight of residues. The percentage of silt was

calculated by subtracting the sum of percentage of all fractions (sand

plus clay) from 100. The results are recorded in the table IV.

Determination of plH:

The pH of soil was determined with Elico pH meter (model LI 120)

using glass and saturated calomel electrodes assembly. A 10 g soil was

dispersed in 50 ml distilled water (1:5, soil: water) to prepare the

suspension and was used to measure the pH of the soil. The result

obtained is recorded in table IV.

Determination of electrical conductivity:

The electrical conductivity of the soil was measured by using Philips

Conductivity Bridge with dip type cell at 30 ± 1°C. A 1: 5, soil: water ratio

CffA(Pm(R. I 87

was used to measure the electrical conductivity. The result obtained is

given in table IV.

Determination of organic matter:

The organic matter of the soil was estimated by using Walkley and

Black (1947) method:

Regents required:

Aqueous solution of potassium dichromate (1N).

Cone, sulphuric acid.

Phosphoric acid (85%).

Alcoholic solution of diphenylamine indicator.

Aqueous solution of ferrous ammonium sulphate (N/2).

Procedure:

A 2 g soil sample, air-dried and passed through a 100 mesh sieve

(BSS), was taken in 500 ml conical flask. 10 ml of 1 N potassium

dichromate solution and 20 ml of concentrated sulphuric acid were added

to it. The flask was shaken vigorously several times, allowed to stand for

30 minutes and thereafter 200 ml of distilled water, 10 ml of phosphoric

acid and 1 ml of diphenylamine indicator were added to it. The excess of

unreacted potassium dichromate was titrated against standard N/2 ferrous

ammonium sulphate solution till the violet colour changed to purple and

finally to green. Reagent blank determination was also carried out in the

CHWPE'R. I

same way. From the volume of dichromate solution used for oxidation,

organic carbon was calculated by using the expression:

^ u /o/x (Blank titre-Actual titre)x0.003xNx 100 Organic carbon (%) =-^ '-

weight of dry soil in gm

where,

N is the concentration of ferrous ammonium sulphate.

The value of organic carbon was converted to organic matter by

multiplying with the factor 1.724. The result obtained for the percentage of

organic matter is recorded in table IV.

Determination of cation exchange capacity:

Cation exchange capacity was determined by the Jackson (1958)

method as:

Reagents required:

0.05N Hydrochloric acid.

IN Sodium acetate solution (pH 5).

IN Calcium chloride solution.

EDTA solution.

1% Alcoholic solution of eriochrome black "T".

Buffer solution of (pH 10).

2% Aqueous solution of sodium cyanide.

CHWVEfi 1

Procedure:

A 5 gm dried soil sample was taken in 100 ml conical flask. The

soluble salts were washed out by treating the soil with 0.05N HCI and then

washing with distilled water. It was further treated with IN sodium acetate

of pH 5.0 for 30 minutes with intermittent stirring in a 100 ml conical flask.

The acidified sample was given five washings with IN standard calcium

chloride solution. The excess salts were removed by washing with 80%

acetone until the excess CaCl2j removed, as indicated by a negative

AgN03 test for chloride ion in the last washing. Finally calcium was

exchanged from Ca - soil by sodium ions through five washings with a

neutral 1N sodium acetate solution. The washing was collected and

utilized in the determination of exchanged calcium ions by titrating it with

a standard solution of EDTA, using 10 ml of the NH4CI-NH4OH buffer of

pH 10 and eriochrome black "T" indicator in presence of 1 ml of 2 percent

NaCN solution as masking agent. A reagent blank was also determined

simultaneously to avoid any error due to interfering ions. The blank

reading was subtracted from the reading of calcium determination. From

the volume of the EDTA solution used, the value of cation exchange

capacity was calculated by using the following expression:

Exchange capacity (meq. 100g^ soil) = "^ xNxlOO Weight of the soil in gm

where,

V is the volume of the EDTA solution and N is the normality of EDTA

solution. The result of the cation exchange capacity of soil is recorded in

table IV.

CffA'PM'R. I 90

Determination of excliangeable cations:

The exchangeable cations of soil were determined as follows:


Buckner funnel.

Systrinicks flame photometer.

Aqueous solution of ammonium acetate (1N).

Nitric acid (6N).

30% Hydrogen per oxide.

6N Hydrochloric acid.

Buffer solution {pH-10).

Eriochrome black "T".

Mureoxide indicator.

10% Potassium hydroxide.

EDTA solution.

Standard solution of Na and K.

Procedure:

A 50 g soil sample, air-dried, grounded and sieved through 100

mesh sieve (BSS), was taken into a 250 ml conical flask and then 100 ml

1N NH4OAC solution was added to it. The contents of the flask were

shaken for 20 minutes and allowed to stand overnight. The soil contents

were then transferred into a Buckner funnel, in which moist whatman filter

paper No. 42 was seated using a gentle pressure. The soil was leached

aoi(pmii I 91

with an additional 400 ml of NH4OAC. The filtrate, containing NH4OAC

extract of the soil, was evaporated to dryness on a steam plate. The dark

coloured residue, containing organic matter^was treated with 2 ml of 30%

H2O2 and 2 ml of 6N HNO3 and heated to dryness on a steam plate. The

dried organic matter free residue was then dissolved in 10 ml of 6N HCI

and diluted with distilled water. It was filtered through Whatman filter

paper No. 42 and the volume was made up to 100 ml. This solution was

2+ 2+ + +

used for the determination of exchangeable Ca , Mg , Na and K in soil.

Exchangeable calcium plus magnesium was estimated with 10 ml of

above solution by EDTA titration, using a half test tube of buffer solution

(pH-10) and 4-5 drops of eriochrome black "T" indicator. Calcium was also

estimated separately using mureoxide indicator with 10% KOH as

recommended by Jackson (1958). The volume of EDTA solution for

magnesium was calculated by subtracting the volume for calcium from the

volume of calcium plus magnesium used. The exchangeable sodium and

potassium was estimated in the above solution using "Systronicks" flame

photometer. The amount of exchangeable Ca *, Mg , Na*, and K* are

recorded in table IV.

Determination of mobiiity of amino acids by soil thin-

layer chromatography:

The mobility, in terms of Rf value, of amino acids through soil was

determined by soil thin-layer chromatography:

afA<pm^ I 92

Table IV

Physico -- chemical properties of HIrapur fine sandy loam soil

Parameters

pH (1:5, soil: water)

Electrical conductivity (dS m" )

Mechanical composition (%)

Sand

Silt

Clay

Soil organic matters (%)

Cation exchange capacity

(meq. 100 g"'' soil)

Exchangeable cations

(Cmol (p+) kg"^ soil)

Na*

K*

Ca^*

Mg2*

Values

8.5

0.9000

63

24

13

0.41

16.3

1.0

0.5

3.5

1.5

CKA'Pmii I 93


TLC applicator with adjustable tliickness.

Uniform glass plates (20 x 4cm).

Glass chamber (Jars of 25 x 10cm size) with covers.

Micropipette.

Glass sprayer.

Oven.

100 mesh sieve (BSS).

0.01 M Aqueous solutions of lysine, glycine, alanine,

phenylalanine and valine.

0.2% Alcoholic solution of ninhydrin (w/v).

Pesticides (parathionmethyl, dimethoate, carbendazim,

chlorpyrifos, endosulfan and methyldemeton).

Procedure:

Preparation of plates:

For the measurement of mobility (Rf value), the soil sample was

dried, grounded and passed through a 100 mesh sieve (BSS) to get

uniform particle size. The soil was slurried in distilled water containing

varying amount (0, 25, 50, 75 and 100 mglOOg""" soil) of pesticides viz.

parathionmethyl, dimethoate, carbendazim, chlorpyrifos, endoslufan and

methyldemeton. The soil slurry was thoroughly mixed so as to get a

homogenous mixture in each case and was then applied on glass plates

Cf€i<FPE<Ii I 94

(20 X 4cm) with the help of an applicator to have an uniform layer of 0.5

mm thickness. The coated plates were air dried at room temperature (30+

3°C). Two lines were scribed at 4 and 14 cm from base on all plates so as

to allow a running distance of 10 cm for the study.

Loading of amino acids and deveiopment of plates:

A 15 fxl amount of the respective 0.01 M amino acid solution was

applied on the base line of the soil coated TLC plates as a single

application with the help of a microplpette. Plates were then developed in

glass chamber using distilled water as a developer up to a distance of 10

cm as indicated by the upper line on TLC plates by ascending

chromatographic technique. To prevent disintegration of soil in contact

with water, wetted stripes of filter paper (about 2.5 cm wide) were

wrapped around the bottom of each plate before their development.

Drying and detection of ctiromatograms:

The developed plates were air-dried. Amino acids were detected by

spraying a 0.2% alcoholic solution of ninhydrin (w/v) and keeping them in

an oven at 70-80°C for 10-15 minutes till appearance pink-coloured spots.

These spots were found to be stable for several days.

Measurement of Rf values:

Rf value of amino acids was measured by the following relation:

R value = ^^°^^^^ distance moved by the amino acid ' 10

Results are recorded in fig. 6 and table V.

impT^<K J 95

METHYL PARATHION DIMETHOATE CARBENDAZIM

1

0 9

08

07

0 6

0 5

0.4

(A

n >

1

0.9

0.8

0 7

0.6

0.5

0.4

0 3

0.2

0.1 •

0

CHLORPYRIFOS

1

0 9

0.8 ^

07

0.6 \

05

0.4

0.3

0.2

01

0

1

0.9

0.8

0.7 H

0 6

0.5

0 4

0.3

0 2

0.1

0

50

ENDOSULFAN

100

50 100 50

1

0.9 •

0.8

0.7 •

0 6

0.5-

0.4 •

0.3

0.2

0 1

0

50

METHYL OEMETON

100

100 50 100

Concentration of pesticides (mg 100 g' soil)

Mobility of amino acids through soil amended with some pesticides.

Lysine Glycine -A-Phenylalanine -x-Alanine -«-Valine

Fig. 6

CWi<Pl^lLl 96

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CKA<Pm^ I 97

RESULTS AND DISCUSSION:

The mobility of amino acids (Fig.6 Table V) in natural soil followed

the order: valine > alanine > phenylalanine > glycine > lysine. This trend

is in accordance with the increasing order of their molecular masses,

except in case of phenylalanine and lysine. The lower mobility of

phenylalanine as compared to valine, may be due to its lower solubility

(Table VI) in water. On the other hand, the lowest mobility of lysine may

be due to its higher adsorptlve tendency over soil colloids, in which the

chemical structure plays a decisive role in its chemical behaviour as:

COO coo + I Soil pH 8.5 I

H 3 N - 9 - H ^- • HpN-C—H ^ 1 -H+ 2 I

(CH2)4 (CH2)4

NH3 NH3

The presence of two protonated amino groups in neutral medium, out of

which, one still remains in the cationic form even in the basic medium is

most probably the cause of the more adsorptlve behaviour of the molecule

over negatively charged sites of the soil colloids with strong electrostatic

force, resulting in its lowest mobility. Its adsorption on soil colloids can be

illustrated as: NHo NH

.^ + H 3 ^ - ( C H 2 ) 4 - C - C 0 0 - — I . | ^ L > " N 3 H - ( C H 2 ) 4 - C - C 0 0 Soil colloids

H H

cHynpim^i 98

Table-VI Structures and characteristics of amino acids

Amino

acids

Lysine

Glycine

Plienyl alanine

Alanine

Valine

Molecular

masses

1.46.6

75.07

165.00

89.10

117.00

Structures^

COO"

H S N ' - C - H

(CH2)4

NH3 +

coo" 1 1

H 3 N ' - C - H

H

coo' 1

H3N' - C - H

CH2

6 coo" 1 1

H3N' - C - H 1

CH3

coo" 1

H3N' - C - H

H3 C CH3

Isoelectric

points" (pi)

9.59

5.97

5.48

6.00

5.96

Solubility'

(in g 1 0 0 ' )

ml water

Very soluble

24.99

2.96

16.51

8.85

Nature^

Positively

charged

Polar but uncharged

Nonpolar (hydrophobic)

Non polar (hydrophobic)

Non polar (hydrophobic)

a. Lehninger, A .L . ; Nelson, D.L. and Cox, M.M. (1993) . Principles of Biochemistry, 2"" ed.; CBS Pub. and Distributor, Nagpur, p. 115.

b. Greenste in , J .P. and Wini tz , M. (1961) . Chemistry of the Amino Acid; Vol. I, John, Wiley and Sons Inc. New York, London, p. 486

c. Gross, D. and Grodsky, G. (1955) . J. Am. Chem. Soc. 77: 1678.

cK;4.(Fm^ I 99

The mobility order: valine > alanine > glycine can be explained on the

basis of their increasing polarity order. The structure of valine, alanine,

and glycine (Table VI) contain secondary, primary and no alkyl groups,

respectively at their one end of the molecule, capable of inducing

electrons (+1 effect) towards the carbon attached with -NH3 and -COO

groups in these amino acids. As a result, this part becomes rich in

electron density but, poor in positively charged centre with diminishing

cationic tendency in the increasing order of the number of methyl groups

attached to these amino acids. The decreased cationic and increased

anionic tendency may compel these molecules to follow the above mobility

order in soil against the negatively charged sites over soil colloids.

The effect of pesticides on the mobility of amino acids (Fig.VI and

Table.V) was found to have a decreasing trend in case of lysine, alanine

and valine in the order: parathionmethyl > dimethoate > carbendazim >

chlorpyrifos > endosulfan > methyldemeton whereas, increased mobility of

phenylalanine has been observed in the reverse order throughout the

entire range of pesticides (upto 100 mg lOOg'"" soil). On the other hand,

the mobility of glycine increased at lower concentration and thereafter; it

declined at higher concentration of pesticides. The varying mobility trend

of amino acids in various pesticides systems can be understood by

considering the chemical behaviour as supported by their structures:

CWl<FPE^ I 100

C H 3 O . II / ^ \ ^O

Parathionmethyl

H O

H O • OCHc

CHo—N-C—CHo—S-P^

^N—N-C-OCH3 N H

Carbendazim

CI

CI

Dimethoate

S

'^^ 0 - P .

CI

Chlorpyrifos

-OCH;

^OCH-:

CI

Endosulfan

CH3-CH2-S-CH2-CH2-S-P:

Methyldemeton

• OCHc

"OCHc

The maximum reduction in the mobility of lysine, alanine and valine in

parathionmethyl-soil system may be due to the presence of nitro (-NO2)

group which,is capable of forming the strong hydrogen bond with amino or

protonated amino group, in amino acids at soil pH 8.5 that can be

represented as:

-N '

Sofl-parathlonmethyl

-!-N '

•H N Rest part of the amino acid

Hydrogen bonding

H

• H — - U -

Soii-parathionmethyl H

-Rest part of the amino acid

The capacity of forming the hydrogen bonding is governed by the number

of electronegative nitrogen or oxygen atoms in these pesticide molecules.

a{ji<Fm^ I 101

that are capable of retaining these amino acids in the order:

parathionmethyl > dimethoate > carbendazim > chlorpyrifos > endosulfan

> methyldemeton. The mobility of amino acids has been found to

decrease in these systems with increasing doses of pesticides. However,

in the case of phenylalanine it seems to be governed by its incapability for

the formation of hydrogen bonding on increasing concentrations of

pesticides, due to the presence of bulky phenyl group, that pose a steric

hindrance resulting the increase of its mobility. On the other hand, the

polarity of glycine enhances its mobility at lower concentrations

(parathionmethyl and dimethoate upto 50 mg and carbendazim,

chlorpyrifos, endosulfan and methyldemeton up to 75 mg 100g"^ soil) due

to its incapability in the formation of hydrogen bonding but, at higher

pesticide concentrations the increased number of electronegative oxygen

and nitrogen atoms of pesticides in soil system are capable of diminishing

the mobility through hydrogen bonding.

cKA-inm^ I |io2

^ferences

• Alexender, M. (1977). Introduction to Soil Microbiology, 2" ^ ed.; Wiley and Sons, New York.

• Apple, T.; Pfanschilling, R. and Xu, F. (1999). J. Plant Nutr. Soil Sci., 162: 615.

• Bremner, J.M. (1952). J. Sci. Food Agr. , 3:497.

• Camazano, M.S. and Martin, M.J.S. (1987). Applied Clay Res., 2:155.

• Devine, M.D.; Duke, S.O. and Fedtke, C. (1993). Physiology of Herbicide Action; PTR Prentice Hall, Englewood Cliffs, New Jersey, p.251.

• Greeves, M.P.; Davies, H.A.; Marsh, M.A.P. and Wingfield, G.J. (1976). CRC Crit. Rev. Micro., 5 : 1

• Jackson, M.L. (1958). Soil Chemistry Analysis; Prentice-Hall, Inc. Englewood Cliffs, N.J.P., 84.

• Khan, S.U. and Khan, N.N. (1986). So/7 Sci., 142: 214.

• Kishore, G.M. and Shah, D.M. (1988). Anna. Rev. Biochem., 57:629.

• Kaneko, T.; Ohkawa, H. and Miyamoh, J. (1978). J. Pestic. Sci., 3:43.

• Li, R.; Jiang, S. and Zhao, J. (1999). Huaxue Shijie, 40: 47.

• Martin, J.P. (1964). Residue Rev., 4: 96.

• Mehta, S.L.; Lodha, M.L. and Sane, P.V. (1993). Recent Advances in Plant Biochemistry; Indian Council of Agricultural Research, New, Delhi, p.453.

• Mody, N.I. and Munshi, C.B. (1990). J. Agril. Food Chem., 38:636.

• Parr, J.F. (1974). So/7 Sci. Soc. Am.; Madison, U.S.A., p. 315.

• Patil, P.S.; Gadkari, M.P. and Kulkarni, K.M. (1992). Environ. Ecol., 10: 758.

cKwm<s^ I 103

• Piper, C.S. (1950). Soil and Plant Analysis; University of Adelaide, Australia.

• Rending, V.V. (1951). Soil. Sci., 71: 253.

a Scheffer, F.; Der, O.G.; Scickstoff, des B. and Siene, V. (1958). Handbuchde Pflanzenphysysiologie, 8: 179.

• Shaner, D.L. (1989). Rec. Adv. Photochemistry, 23: 227.

• Singhal, J.P.; Khan, S.U. and Bansal, V. (1978). Proc. Indian Natn. Sci. Acad., 44-267.

• Sowden, F.J. (1956). So/7 Sci., 82: 491.

• Stevenson, F.J. (1956). So/7 Sci. Soc. Am. Proc, 20:201.

• Sumida, N.; Yamamoto, K. and l\/latsuzaka, Y. (1993). Nippon Dojo Hiryogaku Zassi, 64: 289.

• Talibuddin, O. (1955). Trans. Faraday Soc, 51: 582.

• Theng, B.K.G. (1974). The Chemistry of Clay Organic Reaction; Ch. IV; Adam-Hilger, London, p. 159.

• Wainwright, IVI. and Pugh, G.J.F. (1975). Ibid, 7: 1.

• Waksman, S.A. (1952). So/7 Microbiology; VII, John Wiley and Sons Inc., p.168.

• Waksman, S.A. and Lomanitz, S. (1924). J. Agr. Res., 30: 263.

• Waksman, S.A. and Starkey, R.L. (1932). J. Bact., 23: 405.

• Walkley, A. and Black, C.A. (1947). So/7. Sci., 63:251.

• Yanron, S. (1978). So/7 Sci., 125: 210.

CHAPTEE II

EFFECT OF FLY ASH ON THE MOBILITY

OF AMINO ACIDS THROUGH SIX TYPICAL


chapter II

INTRODUCTION

c > ^ ^ ccording to the order of genesis of the principle soil

C^ \y types, Aligarh district has been grouped into six typical

soil zones (Agarwal and Mehrotra, 1951) viz. Ganga

loamy sand (Type I), Aligarh loam (Type II), Aligarh clay loam (Type III),

Aligarh sandy loam (Type IV), Yamuna silty clay loam (Type V) and

Yamuna sandy loam (Type VI). Due to the little leaching and

considerable accumulation of salts on the surface of these soils, the

agricultural production of this district is low. Waste products such as, fly

ash, sewage sludge, industrial wastes, etc., can be tried as soil

amendments for improving crop yield. Since, the waste products

contain certain organic and inorganic components, which are required

by living organisms. The fly ash, released from thermal power plants,

locomotives and other industries, is based on fossil fuels (Rathor et al.,

1985; Roshi, 1981) and therefore, it has a great potential to be utilized

as a nutrient, for healthy plant growth (Plank, 1947; Adriano et al.,

1978; Mishra and Shukia, 1986). But, the long-term use of fly ash is

harmful to the living organisms (Majumdar and Mukharjee, 1983;

C3Ci<P^^n I 105 I

Pichtel, 1990; Pichel and Hayes, 1990; Udaipur et al., 1998) due to the

accumulation of some toxic metals (Chandra, 1997; Bell and Phung,

1979) in soils. Fly ash changes the physical and chemical properties of

soils (Khan et al., 1996; Hill and Lamp, 1980; Page et al., 1979) that

may affect the translocation of useful compounds such as, proteins,

amino acids, etc., which play an important role in the biochemical

processes (Theng, 1974). The free amino acids in soil are present in

very small quantities (Paul and Schmidt, 1960), but there is good

evidence that 24 to 37 percent of the nitrogen in soil is in the a - amino

forms, which suggests the proteinaceous character of this fraction

(Firman, 1964; Bremner, 1958; Bremner and Shaw, 1954). The

macromolecule proteins and other organic matter are broken down into

amino acids (Wakswan and Starkey, 1932; Waksman and Lomanitz,

1924). They may adsorb onto soil colloids or translocate into plants

(Eyiar and Schmidt, 1959) on the basis of soil condition.

The use of fly ash has been recommended by a number of

agricultural scientists (Sarangi and Mishra, 1998; Khan and Khan,

1996; Sikka and Kansal, 1995; Sarangi et a!., 1997), but still more work

is needed to ascertain the full impact of fly ash while using in

agricultural soils.

cm<pm^ii 106

EXPERIMENTAL

Six typical soil samples, collected from different parts of Aligarh

district (India), were used to conduct the study. The physico-chemical

properties of soils were determined by standard methods as described

in chapter I. Results are given in table VII.

Fly ash was collected from Thermal Power Station, Kasimpur in

Aligarh district (India) and was sieved through 100 mesh sieve (BSS).

The pH, E. C, mechanical analysis, organic carbon and available

potassium of fly ash were determined by the standard methods used for

soil in chapter I.

Available nitrogen and phosphorus components of soils and fly

ash were determined by the following methods:

Determination of avaiiable nitrogen:

Available nitrogen was determined by the method of Hesse

(1979) as follows:

Determination of ammonium nitrogen:

Reagents required:

• Potassium chloride solution (2 IVI): A 150 g KCI was

dissolved in 800 ml distilled water and boiled with 10 g

C3Ci<FPE^II I 107

magnesium oxide for about 15 minutes, until any ammonia

present was expelled. The solution was then cooled and

filtered before making the final volume.

• Magnesium oxide: Magnesium oxide was heated at 650 C

for two hours in an electric muffle furnace and allowed to

cool in a desiccator over solid KOH and stored in a tightly

stoppered glass bottle.

• IVIixed indicator: Mixed indicator solution was prepared by

dissolving 0.1 bromo cresol green and 0.07 gm methyl red

in 100 ml of ethanol.

• Boric acid solution: A 20 g of boric acid was dissolved in

900 ml of hot distilled water. After cooling the solution, 20

ml mixed indicator solution was added to it. Thereafter, 0.1

M NaOH solution was added drop wise to it until the colour

changed to reddish purple.

• M/70 hydrochloric acid solution:

Procedure:

5 g soil or fly ash was taken in 100 ml glass stoppered conical

flask. A 50 ml of 2 M KCI solution was added in it. The flask was then

shaken vigorously for an hour and the soluble contents were extracted

through Whatman filter paper No. 42. A 10 ml solution from extracts

was taken in distillation flask and diluted with 50 ml distilled water. A

cm<P^<iLn 1081

0.5 g magnesium oxide was added througli long-stemmed funnel into

the flask. Ammonia was distilled in to a solution of 5 ml boric acid

containing mixed indicator tlirougli a condenser until tlie final volume of

the distillate was reached to about 30 ml. The boric acid was then

titrated with M/70 HCI by using a micro burette until the green colour

changed to pink. A blank was also run in the same way. The available

ammonium nitrogen was calculated using the relation:

1 ml of M/70 HCI = 0.2 gm of nitrogen.

The results obtained, are recorded in table VII (soil) and VIM

(FA).

Determination of nitrite nitrogen:

Reagents required:

• All the reagents used in the determination of ammonium

nitrogen.

• Devarda alloy: Finally grounded and passed though a 0.15

mm sieve.

Procedure:

After distillation of NH4 *-N, the stopper of the distillation flask

was removed. Therefore, a 0.2 g of Devarda alloy and 50 ml of distilled

water were added to it. The distillation flask was stoppered and then

ammonia was distilled in a fresh portion of boric acid solution. About 30

CWfPffifll// 109 I

ml distillate was collected and titrated with M/70 HCI. The value of

nitrite nitrogen plus nitrate nitrogen was calculated in the same manner

as described in the determination of ammonium nitrogen. The value of

nitrite nitrogen was obtained by subtracting the value of nitrite nitrogen vtt i

from it. The results are given in table VII (soil) and (FA).

Determination of nitrate nitrogen:

Reagents required:

• All the reagents as used in the determination of nitrite

nitrogen.

• A 2% sulphamic acid solution.

Procedure:

The experimental procedure was partly the same as described in

the determination of ammonium nitrogen. After the removal of

distillation flask, one ml of a 2% aqueous solution of sulphamic acid

was added to it. The distillation flask was swirled for few seconds to

destroy the nitrate. Accordingly, 0.2 g Devarda alloy and 50 ml distilled

water were added in distillation flask. It was stoppered and ammonia

was distilled with a fresh portion of boric acid solution. About 30 ml

distillate was collected in each case and titrated with M/70 HCI. The

value of nitrate nitrogen was calculated in same manner as described in

the determination of ammonium nitrogen. The results are given in table

VII (soil) and VIM (FA).

cHMim.li I no I

Determination of available phosphorus:

Available phosphorus was determined by the method of Olsen

(1954) as:


• Bausch and Lomb spectronic "20".

• 0.5 M sodium bicarbonate solution (pH 8.5).

• Darco G 60, phosphorus free charcoal.

• Ammonium molybdate solution: It was prepared by

adding 15 g (NH4)2Mo04 and 30 ml cone. HCI in 650 ml

warm distilled water.

• Stannous chloride solution: For this solution 10 g

SnCl2.2H20 was dissolved in 25 ml cone. HCI, then 0.5 ml

of this solution was dilute with 66 ml distilled water.

• Phosphorus standard solution:

Procedure:

A 5 g of soil or fly ash sample was taken in 150 ml conical flask.

One teaspoon of carbon black was added in conical flask. Then it was

treated with 100 ml of extracting solution (0.5M NaHCOa, pH 8.5).

Contents of the flask were shaken for 30 minutes on a mechanical

shaker and then it was filtered through Whatman filter paper No. 42. A 5

ml of the clear extract was taken in 25 ml measuring flask and 5 ml of

ammonium molybdate was added to it. The neck of the flask was

washed with

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CWPTE'till 112

Table VIII Physico-chemical properties of fly ash

Parameters

pH, (1 : 5, FA:water)

Electrical conductivity (dS m )


Sand

Silt

Clay

Organic carbon (%)

Available nitrogen (mg kg'^ fly ash)

NH4-N

NO3-N

NO2-N

Available phosphorus (mg kg"^

Available potassium (%)

fly ash)

Values

6.72

0.71

25.50

72.00

0.60

0.60

30.40

24.80

3.20

470.00

0.7320

CHMTfE^ll I 113

distilled water. After shaking this solution, 1 ml of stannous chloride was added

to it and then, volume was made up to the mark. Blue colour was then

developed. A blank was prepared in the same way. The absorbance of the

solution was recorded with Bausch and Lomb using Spectronic '20' at a

wavelength of 660 \i using a red filter. The results are given in table VII (soil)

and VIII (FA).

Determination of tlie mobility of amino acids through

soil thin-layer chromatography:

Effect of fly ash on the mobility (Rf value) of amino acids in six typical

soils of Allgarh district, using soil thin-layer chromatography, was investigated

as follows:


TLC applicator with adjustable thickness.

Uniform glass plates (20 x 4 cm).

Glass chambers (jars of 25x10 cm size) with covers.

Micropipette.

Glass sprayer.

A 100 mesh sieve (BSS).

Oven.

0.01 M Aqueous solutions of lysine, glycine, alanine and

valine.

0.2% alcoholic solution of ninhydrin.

CMMnm^n I 114 I

Procedure:


The soil samples were dried, crushed and sieved through a 100

mesh sieve (BSS). For the study of the effect of fly ash on the mobility

of amino acids, these soil samples were amended with varying

concentration of fly ash (0, 5, 10, 15 and 20g kg'"" soil), and were used

as static phase. Slurries of these amended soils were prepared in soil

water ratio of 1:2 and were applied on glass plates (20 cm x 5 cm) with

the help of an applicator to get uniform layer of 0.5 mm thickness. Two

lines were scribed at 4 and 14 cm from the base at the surface of all

plates.

Application of samples and development of plates:

An aqueous solution of 10 )LII of each amino acids (0.05 M) were

spotted at the base line on separate TLC plates with the help of

micropipette. Plates were developed in closed chamber using distilled

water as a mobile phase to the distance of 10 cm (Helling and Turner,

1968; Singhal et al., 1978) by ascending chromatography technique. To

prevent disintegration of soil In contact with water, wetted strips of filter

paper (about 2.5 cm wide) were wrapped around the bottom of the

plates before the development.

Drying and detection of chromatograms:

Developed plates were air-dried. Visualization of amino acids on

CHMl^^II 115

TLC plates was done by spraying 0.2% alcoholic solution of ninhydrin

(w/v) and heating in an oven at 70-80°C for 10-15 minutes. Amino acids

were detected as brick red coloured spots, which were stable for

several hours.


The mobility, in terms of R( value, was measured by the formula:

• Frontal distance moved by amino acid K, = —

f 10 Results are given in fig. 7 and table IX.

CK^rTLH 11 116

(0 0)

n >

1.2

08

0 6 -

0 4

0 2

1

0.9 •

0.8

0.7

0.6 •

0.5

0.4

0.3 •

0.2

0.1

0

TYPE I

10

TYPE VI

10

20

20

1.2

0.8 -

0.2

0.9

0.8

0.7 •

0.6

0.5 •

0.4

0.3 •

0.2

0.1

0

TYPE IV

10

TYPE III

10

20

20

1

0.9

0.8 -I

0.7

0.6

0.5

0.4

0.3

0.2

0.1

0

0.9

0.8

0.7

0.6

0.5

0.4

0.3

0.2

0.1

0

TYPE I

10 20

TYPEV

10 20

Cone, of fly ash (g kg'^ soil)

Effect of fly ash on the mobility of amino acids through six typical soils of Aligarh district

Lysine Glycine ' Alanine -Valine

Fig. 7

OfA^Pl^^II 117

(0 O)

a

o w ><

'w

O)

2 <fl

(0

I-

o ra o c 1 (0

o

o E

c o

(0

O

liJ

—

o (A 1

O)

O) • ^ ^

JC in (0

o c _o CO ^ c 0) o c o o

o OJ

in ^

o ^

If)

o

o CM

lO ^

o

in

o

o (M

in

o ^

in

o

0) 3

n > •

It'

z 0)

1-

z 0) a >> 1-

a> a 1-

O (0

E o < «

in

q d

q d

00

q d

o T-

d

T-

d

a> q d

• ^

d

CNJ

d

CN T—

d

(O

d

o

d

CM T—

d

m ^ d

in '

d

CO CNi

d

c

_J

CN N-

d

CO

q d

o q d

r q d

o q d

in N-

d (N h--

d

o

d

N-

q d

o d

in q d

q d

CO

q d

in N.

d

CO 00

d 0)

"o O

o q d

00

q d

h-d

CO h-.

d

N-

d

CO

q d C3> N.

d

00 1 d

CM

q d

CO q d

q

00

q d

in

q d

CNJ

q d

CN

q d

0)

'c CO <

in

q d

eg q d

in S; d

00 N-

d

q d

CO

q d o q d

N-q d

CO

q d

o q d

q

oo q d

-f

q d

o q d

CO

q d

"cc >

> 0) Q.

H

>

a 1-

>

0) a >> 1-

00

q d

o T-

d

o T~

d

CVJ T-

d

d

in

q d 00

q d

C3) q d

CJ)

q d

OJ q d

o

d

CM T"

d

CM — d

-"I-

d

CJ) T-

d

0)

_l

CO

q d

CO N-

d

CO

q d

o q d

q d

CO N-

d h-

q d

o q d

o q d

in q d

d

h-

d

CM N-d

CJ)

q d

N-

d

0)

c

O

CM

q d

o q d

CO 1 d

r d

CM

q d

CO

q d in h-

d

o

d

CO

q d

^

d

CM

q d

c» q d

i

q d

in

q d

o q d

0)

'c CO <

CO

q d

1 q d

CO

q d

C3) h-

d

m q d

in

q d o q d

h-

d

" N. d

00

d

q d

q d

oo q d

00

q d

CO

q d

0)

75 >

aoirpE<iiii 118

RESULTS AND DISCUSSIONS

As is evident from fig. 7 and table IX, that the mobility of amino

acids in six typical soils of Aligarh district followed the order: valine >

alamine > glycine > lysine. The results may be discussed on the basis

of the physical and chemical characteristics of soil and the molecular

behaviour of amino acids in soil medium. The lowest mobility in case of

lysine may be due to the presence of - NH3 group even in the alkaline

range of soil pH < pi (Table VII and VI). Hence, it may be expected to

show highest degree of adsorption and thus, remained bound with

electrostatic force on the negatively charged sites of the soil colloids

as:

cod Soil colloids NH,(CH2)4 C - N H , I 3

H

On the other hand, the presence of single -NH3 group of other three

amino acids get converted into -NH2 group at soil pH > pi. Therefore,

the probability of interaction of these amino acids towards soil colloids

saturated with hydrated or unhydrated metal ions (Mortensen and

Himes, 1964; Baily et al., 1968) may be as:

O" NH2

Soil colloids h M O ^ ~:0 = C -C - H

R

CHMT^'^II

n* . / H H COO' Soil colloids - M oC I 1

H R

where, - R = - H in glcycine , - R = - CH3 in alanine and - R = -CH(CH3)2in valine.

Soil Colloids - M -

O NH2 II I

0 - C - C - H

R -n-1

The mobility of these three amino acids can be explained on the basis of

their molecular make up and capabilities of binding with adsorptive sites

of soil colloids. They were found to remain in the sequence of their

molecular polarity. The highest mobility in case of valine may be attributed

to the presence of the biggest alkyl group that imposes the steric

hindrance in its interaction process with soil colloids. However, glycine is

most polar and exerts greater attractive force on the adsorptive sites

resulting in the decrease of its mobility.

The variation in the mobility trends of amino acids in different soils

followed the order: Type I > Type IV > Type ll> Type Vl> Type lll> Type V

>. This order of mobility directly seemed to be related to their clay

contents (vide table IX). Thus, soil Type I containing lowest percentage of

clay contents appeared to behave the least adsorptive and more porous

due to high-silt sand contents. Hence, it paved the way for amino acids to

cm<pTm.ii I 120

move at faster rate. Similarly, clay contents in soil Type V is highest,

resulting in its lowest mobility.

In the fly ash admixed soil, the mobility of amino acids decreased at

low concentration but,increased at higher concentration (except lysine) in

all amended soil types. The diminished mobility at lower concentration of

fly ash may be attributed to the saturation of clay particles by metal ions,

released from fly ash. Hence, amino acids have greater chance to interact

on newly formed clay-metal complexes. Free carbon and metal oxides of

fly ash (Saxena et al., 1998; De and De, 1994) may also enhance the

interaction processes. Similarly, the porosity and concentration of sand

and silt increases on increasing concentration of fly ash that may raise the

mobility of amino acids. On the other hand, the mobility of lysine has been

found to decrease on increasing concentration of fly ash. As

concentrations of fly ash increase, carbon content containing adsorptive

sites also increased. Thus lysine should have greater chance to adsorb on

these adsorptive sites, resulting in decreased of its mobility.

CHMim^II 121

<Kfferences

• Adriano, D.C.; Woodford, T.A. and Ciravolo, T.G. (1978). J. Environ. Qual., 7:416.

• Agarwal, R.R. and Mehrotra, C.L. (1951). So/7 Survey and Soil Work in U.P.; Vol. I - IV, Supdt. Printing and Stationary, U.P., Allahabad.

• Bailey, G.W.; White, J.L. and Rothberg, T. (1968). Soil Sci. Soc. Amer. Proc, 32: 222.

• Bell, H.T. and Phung, L.T. (1979). J.Environ. Qual., 8:171.

• Bremner, J.M. (1958), J. Sci. Food Agr., 9 : 528.

• Bremner, J.M. and Shaw, K. (1954). J. Agr. Sci., 44: 152.

• Chandra, K. (1997). Indian J. Agric. Chem., 38: 87.

• De, Anil K and De, Arnab K. (1994). Journal lAEM, 21 : 36.

• Eyiar, O.R. and Schmidt, E.L.(1959). J.Gen. Microbial., 20: 473.

• Firman, E.B. (1964). Chemistry of the Soil, 2" ^ ed.; Oxford and IBH Publishing Co., New Delhi, p. 226.

• Helling, C.S. and Turner, B.C (1968). Science, 162: 562.

• Hill, M.J. and Lamp, L.A. (1980). Aust. J. Exptl. Agric. Animal

Husb., 20 : 377.

• Khan, M.R. and Khan M.W. (1969). Environ. Poll.,92: 105.

• Khan, S.U.; Begum, T. and Singh, J. (1996). Indian J. Environ. HLTH, 38 :41 .

• Majumdhar, A. and Mukharjee, S.N. (1983). Fuel Sci. Technol., 2: 80.

• Mishra, L.C. and Shukia, K.N. (1986). Environ. Pollut. {Series /A), 42:1.

• Mortesen, J.L. and Himes, F.A. (1964). Chemistry of the Soi, 2"^ ed.; Oxford and IBH Publishing Co., New Delhi, p. 206.

a Page, A.L.; Elseewi, A.A. and Straughan, I.R. (1979). Resid. Rev., 71 : 83.

cm<PtE<fin 111

• Paul, E.A. and Schmidt, E.L. (1960). Soil Sci. Am. Proc, 24: 195.

• Pichtel, J.R. (1990). Environ. Pollut., 63: 225.

• Pichtel, J.R. and Hayes, J.M. (1990). J. Environ. Qua!., 19: 593.

• Plank, O.C. (1947). Diss. Abstr. Int., B, 34: 4156.

u Rathor, H.S.; Sharma, S.K. and Agarwal, M. (1985). Environ. Pollut. {Series B),^0•.2A9.

• Roshi, S.S. (1981). Building Material from Indian Fly Ashes, Central Building Research Institute, Roorkee (UP.).

• Sarangi, P.K. and Mishra, P.C. (1998). Res. J. Chem. Environ., 2:7.

• Sarangi, P.K.; Mishra, T.K. and Mishra, P.C. (1997). Indian J. Environ. Res., 1: 17.

• Saxena, M.; Ashokan, P. and Chauhan, A. (1998). Clay Res., 17: 109.

• Sikka, R. and Kansal, B.D. (1995). Broresource Tech., 51:199.

• Singhal, J.P; Khan, S.U. and Bansal, V. (1978,b). Proc. Indian National Sci. Acad., 44: 267.

• Theng, B.K.G. (1974). The Chemistry of Clay Organic Reactions; Adam-Hilger, London, p. 159.

• Udalpur, S.K.; Soni, P. and Thetwar, L.K. (1998). Res. J. Chem. Environ., 2: 51.

• Waksman,S.A. and Lomanitz, S. (1924). J. Agric. Res., 30: 263.

• Waksman, S.A. and Starkey, K.L. (1932). J. Bact., 23: 405.

CHAPTEE III

MOBILITY OF TRACE METALS IN SOIL AS

INFLUENCED BY SOME SURFACTANTS

CHapterlll

INTRODUCTION

urfactants are one of the major components of detergents

and household cleaning products, which are used in large

quantities. Long-term treatment with surfactants causes

pollution (Valoras et al., 1976) of water and soil, which affects growth,

development and metabolism of soil microorganisms (Kowalezyk, 1993;

Kozhevin, 1994; Ribionovich et al., 1994), plants (Hall et al., 1980;

Pfleclger et al., 1990) and aquatic animals (Lewis, 1991) and other

organisms (Poremba et al., 1991). They solubilize non-polar plant

substances such as the waxy cuticle or the lipoidal portion of the cell

wall that facilitate the rapid absorption of the toxic along with beneficial

chemicals. They enter into the intercellular spaces and thus adversely

affect the plant growth systems. Further, the surfactants have also

affect on the proteins and amino acids (Husain and Ahmad, 1993). They

possess powerful bactericidal properties and are also known to modify

the physico-chemical properties of soils (Klingman, 1973).

Trace metals have been found to play an important role in the life

processes of plants, animals and microorganisms (Jain, 1974). Among

the trace metals, micronutrients are essential for proper growth of plant

CW'PPEiini 124

whereas certain heavy metals are hazardous for plants (Dijkshoorn et

al., 1979; Khan and Khan, 1983), animals (Underwood, 1971), micro

flora (Doelman, 1978) and soil microorganisms (Brookes and McGrath,

1984; Brookes et al., 1984), as well as for human beings (Schmitt et al.,

1979). Heavy metals may inhibit microbial enzymatic activities and

reduce the diversity of population of microorganisms and soil fauna

(Tyler, 1981; Kuster and Grun, 1984). Reduction in population of soil

organisms by high concentrations of heavy metals has been

documented (Hirst et al., 1961; Mochizuki et al., 1975; Hughes et al.,

1980).

A large number of organic chemicals are either released from

industrial waste disposal and other chemical laboratories or directly

converted from soil organic matters (Artsukevich and Solomon, 1997;

Schwartz et al., 1954; Allison, 1973; Miller and Wolin, 1992; Ravinder et

al., 1994) that are get migrated to agricultural lands. Some of these

chemicals are highly toxic to organisms. Thus, the presence of the toxic

chemicals in soil is an economic loss and a threat to the quality of our

soil surface in relation to agricultural production.

A number of workers have studied the interaction of trace metals

(Khan et al., 1983; Khan et al., 1996) and organic chemicals (Eigen,

1994; Khan et al., 1985; Patzko and Dekany, 1996) in relation to soils.

Meager information is available on the role of surfactants in the

mobilization processes of trace metal through soils with regards to

organic acids as mobile phase.

The present study is an attempt to investigate the mobility of trace

metals through soil amended with surfactants, which may provide the scientific

base to tackle the wide spread problem of pollution.

aci^TiE<iiiii 125

EXPERIMENTAL

The soil (0-30 cm) used in this investigation, was collected from

Agricultural Farm of Aligarh Muslim University, situated in Aligarh

district (U.P.) India. Physcio-chemical properties of the soil were

determined by methods as described in chapter I and results are

recorded in table IV (chapter I).

Determination of mobility of trace metals through soil thin-layer chromatography:

The mobility of the trace metals, in terms of Rf values, was

determined by soil thin-layer chromatography as follows:


The applicator with adjustable thickness.

Uniform glass plates (20 x 4 cm).

Glass chambers (Jars of 25 x 10 cm size).

Micropipette.

Glass sprayer.

!00 mesh sieve (BSS).

Oven.

0.1M Aqueous solutions of Pb, Cu, Ag, Zn and Co nitrates.

0.1 % Alcoholic solution of haemotoxilin (w/v).

0.25M Aqueous solutions of formic, acetic and oxalic acids.

CHA<PtEfiIII 126

• Surfactants: Cetyltrimethylammonium bromide (CTAB), t-octyl-

phenoxypolyethoxy ethanol (Triton x-100) and sodiumdodecyl

sulphate (SDS).

Procedure:


The mobility of trace metals was measured through a sample of

soil having uniform particle size (well grounded and passed through 100

mesh sieve). The soil was amended with varying quantities of

surfactants viz. CTAB, Triton x-100 and SDS containing 0, 25, 50, 75

and lOOmg lOOg' soil. The amended soil was mixed thoroughly to get

a homogenous mixture that was converted into soil slurry with a soil

water ratio of 1:2 in each case. The slurry was coated on glass plate

with the help of an applicator to get uniform layer of 0.5 mm thickness.

These plates were air dried at room temperature, 30 ± 3°C. Two lines

were scribed at 4 and 14 cm from the base of each plate so as to allow

a developing distance of 10 cm for the study.

Application of sample and development of plates:

An aqueous solution of 0.1 M trace metal nitrates of Pb, Cu, Ag,

Zn, and Co were spotted at the base line in a single application with the

help of a micropipette. These dried spotted plates were developed in

closed chamber using distilled water or 0.25M organic acid solution,

viz., formic, acetic and oxalic acids as a mobile phase up to a distance

CHMT^'R.III ni

of 10 cm by ascending chromatography technique. To prevent

disintegration of soil in contact with water, wetted strip of filter paper

(about 2.5 cm wide) were wrapped around the bottom of plates before

the development.

Drying and defection of ciiromatogram:

Developed plates were dried in an oven at 60°C temperature.

Trace metals were detected by spraying a solution of 0 .1% haemotoxilin

in ethanol (w/v) that gave dirty green for Ag, light blue for Co and dark

blue for Zn, Cu and Pb. These coloured spots remained stable for

several hours.


The mobility, in terms of Rf values, was measured by using the formula:

" R L + R T R,-X 0

where,

RL = Leading distance moved by metal.

RT = Trailing distance moved by metal.

(RL + RT) / 2 = The average of leading and trailing distance from the

base line.

Results are recorded in fig. 8-11 and table X-XIII.

OfM^-L'K III 128

i

CTAB

50 100

TRITON x-100

Cone, of surfactants (mg 100 g soil)

Mobility of trace metals in surfactants amended soil

CHyKFTEIilll 129

Table X

Mobility of trace metals in surfactants amended soil

Trace

metals

Pb

Cu

Ag

Zn

Co

Pb

Cu

Ag

Zn

Co

Pb

Cu

Ag

Zn

Co

Rf values

Concentration of surfactants (mg 100g

0

0.07

0.09

0.12

0.14

0.44

0.07

0.09

0.12

0.14

0.44

0.07

0.09

0.12

0.14

0.44

25

CTAB 0.10

0.12

0.15

0.19

0.47

50

0.14

0.16

0.17

0.22

0.50

TRITON X-100

0.10

0.11

0.14

0.18

0.46

SDS

0.05

0.07

0.11

0.12

0.42

0.14

0.13

0.17

0.20

0.51

0.04

0.05

0.09

0.09

0.38

75

0.18

0.20

0.20

0.25

0.54

0.18

0.17

0.20

0.23

0.51

0.03

0.02

0.07

0.07

0.34

•' soil)

100

0.22

0.24

0.25

0.28

0.54

0.22

0.20

0.24

0.26

0.54

0.01

0.01

0.04

0.05

0.34

CK'^rTL'SiiJj 130

w 0)

To >

0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1

0

0.8

0.7

0.6

0.5

0.4

0.3

0.2

0.1

0

0.7

0.6

0.5

0.4

0.3

0.2

0.1

0

CTAB

50

TRITON X-100

100

Pb

Cu

Ag

Zn

Co

50 100

SDS

50 100

Cone, of surfactants (mg 100 g' soil)

Effect of formic acid on the mobility of trace metals in surfactants amended soil

CHMT^^III 131

Table XI

Effect of formic acid on the mobility of trace metals In surfacetent amended soil.

Trace

metals

Pb

Cu

Ag

Zn

Co

Pb

Cu

Ag

Zn

Co

Pb

Cu

Ag

Zn

Co

Rf values


0

0.15

0.19

0.24

0.29

0.64

0.15

0.19

0.24

0.29

0.64

0.15

0.19

0.24

0.29

0.64

25

CTAB 0.18

0.22

0.28

0.32

0.69

50

0.18

0.25

0.29

0.35

0.69

TRITON X-100

0.18

0.21

0.26

0.32

0.67

SDS

0.14

0.17

0.23

0.26

0.62

0.22

0.24

0.28

0.34

0.67

0.11

0.15

0.21

0.23

0.60

75

0.29

0.29

0.34

0.39

0.75

0.24

0.27

0.31

0.36

0.73

0.10

0.13

0.17

0.19

0.58

•' soil)

100

0.31

0.33

0.41

0.43

0.80

0.31

0.30

0.34

0.39

0.77

0.09

0.10

0.14

0.15

0.55

CfOfTTLIlJI] 132

CTAB

w

To >

0.7

0.6

0.5

0.4

0.3

0.2

0.1

0

50 100

TRITON X-IOO

SDS

50 100

Cone, of surfactants (mg 100 g soil)

Effect of acetic acid on the mobility of trace metals in surfactants amended soil

Fig. 10

CKH'PPE'R.III 133

Table XII

Effect of acetic acid on the mobility of trace metals in surfacetent amended soil.

Trace

metals

Pb

Cu

Ag

Zn

Co

Pb

Cu

Ag

Zn

Co

Pb

Cu

Ag

Zn

Co

Rf values


0

0.12

0.15

0.20

0.23

0.59

0.12

0.15

0.20

0.23

0.59

0.12

0.15

0.20

0.23

0.59

25

CTAB 0.15

0.18

0.23

0.26

0.63

50

0.18

0.22

0.27

0.29

0.67

TRITON X-100

0.15

0.17

0.23

0.25

0.62

SDS

0.11

0.13

0.19

0.21

0.57

0.19

0.20

0.26

0.27

0.65

0.09

0.12

0.16

0.19

0.53

75

0.21

0.26

0.31

0.31

0.71

0.23

0.23

0.29

0.30

0.69

0.07

0.11

0.13

0.16

0.53

•' soil)

100

0.25

0.30

0.35

0.35

0.71

0.27

0.27

0.32

0.34

0.73

0.05

0.08

0.11

0.13

0.50

i r ? / K ; ; / 134

CTAB

(A 0)

re >

0.7

0.6

0.5

0.4

0.3

0.2

0.1

0

0.7

0.6

0.5

0.4

0.3

0.2

0.1

0

0.6

0.5

04

0.3

0.2

0.1

0

50

TRITON x-100

100

Pb

Cu

Ag

Zn

Co

50

SDS

100

0 50 100

Cone, of surfactants (mg 100 g' soil)

Effect of oxalic acid on the mobility of trace metals in surfactants amended soil

Fig. 11

CKMT^EIilll 135

Table XIII

Effect of oxalic acid on the mobility of trace metals in surfacetent amended soil.

Trace

metals

Pb

Cu

Ag

Zn

Co

Pb

Cu

Ag

Zn

Co

Pb

Cu

Ag

Zn

Co

Rf values

Concentration of surfactants

0

0.10

0.11

0.14

0.16

0.48

0.10

0.11

0.14

0.16

0.48

0.10

0.11

0.14

0.16

0.48

25

CTAB 0.12

0.15

0.17

0.20

0.53

50

0.16

0.18

0.21

0.22

0.56

TRITON X-100

0.13

0.13

0.16

0.18

0.51

SDS

0.08

0.11

0.13

0.15

0.46

0.16

0.15

0.19

0.20

0.54

0.06

0.09

0.11

0.14

0.44

(mglOOg

75

0.20

0.21

0.25

0.26

0.60

0.19

0.17

0.23

0.24

0.55

0.04

0.08

0.10

0.12

0.41

•' soil)

100

0.24

0.25

0.30

0.30

0.65

0.23

0.21

0.26

0.28

0.63

0.00

0.06

0.09

0.09

0.37

CHMT^fLIII 136

RESULTS AND DISCUSSION

The mobility of trace metals (Fig. 8 and Table X) in natural soil

followed the order. Co > Zn > Ag > Cu > Pb, which is in the sequence of

the reverse order of their binding strengths with soil organic matter

(Stevenson and Ardakani, 1972; Lund ef a/., 1976). The mobility of

trace metals gradually tends to increase in case of CTAB and Triton x -

100 where as, it declined in SDS soil system with the increasing dosage

of surfactant amendments (up to 100 mg 100 g-1 soil) in soil. The

greater mobility of trace metals in case of CTAB as compared to Triton

x-100 may be discussed on the basis of their chemical structures as:

CHo I ^ - ^ N I CH3

Cetyltrimethylammonium bromide (Cationic)

r "3 + pH3(CH2)i5CH2—N—CH^ Br

CH3 CH3

CH3—C—CH2—C—(v />—(OCH2CH2i5pOH

CH3 CH3

t-Octylphenoxypolyethoxy ethanol (Non-ionic)

where x = 10 average

CH3(CH2)iiSO"4Na

Sodiumdodecyl suiphate (Anionic)

The highest mobility of trace metals in case of CTAB may be attributed

to cationic tendency of exchanging ions from the exchangeable sites

(Motlant 1970; Schuiz et al., 1998) and thus to the capability of blocking

CMMTE^III 137

these sites, resulting in trace nnetals mobility at the fastest rates.

However, Triton x - 100 molecule containing hydroxyl group, is capable

of being adsorbed through hydrogen bonding with the hydrated cations

(Motlant 1970; Schuiz et al., 1998), already present over soil colloids.

Thus, it may block the exchangeable sites and facilitating the trace

metals to move at faster rate but, with less than that of CTAB. The

reduced mobility of trace metals in SDS amended soil may be due to its

anionic nature. It induces negatively charged sites in soil and enhances

the interaction of trace metal ions resulting In an Increase in adsorption

sites. This increase in adsorption of these ions results in the decrease

of their mobility.

It is clear from (fig 9-11 and table XI-XIII) that the role of organic

acids (0.25 M), when used as a mobile phase, has markedly

pronounced affect on the mobility of trace metals through soil amended

with surfactants in the order : formic > acetic > oxalic acids. The order

of mobility is inversely related to their pka values (except 1®* pka value

of oxalic acid) and the molecular masses of caboxylates. Formic acid

having less pka value (table XIV) has greater ability to form more stable

metal caboxylates. Hence, metal ions have the least chance to adsorb

on soil colloids. Thus, a greater mobility has been noted in this case.

On the other hand, pka value (2" ^ pka value) of oxalic acid is higher

(table XIV), which forms less stable metal caboxylates. As a result,

trace metals have greater chance to adsorb on soil colloids and

therefore, it showed lesser mobility. Larger size of metal oxalates may

also cause decline in trace metals mobility.

aoiippE<iiiii 138

Table XIV

pka values of organic acids^ and solubilities of trace metal carboxylates'^ and hydroxides^

Organic

acids

Formic Acid

Acetic acid

Oxalic acid

pk

values

3.75

4.05

6.23

&4.19

Trace

metals

Pb

Cu

Ag

Zn

Co

Solubility of metal ca

Metal

formates

1.6

12.5

-

5.2

5.03

Metal

acetates

45.61

7.2

0.72

30.00

very

soluble

rboxylates

Metal

oxalates

0.00016

0.00253

0.00339

0.00079

Insoluble

Solubilities of

metal hydroxides

(g iOOg'^ water)

0.015

insoluble

-

0.00000026

0.00032

Weast, R.C., Astle, M.J. and Beyer, W.H. (1986). Handbook of Chemistry and Physics, 67*" ed; CRC Press, Inc Boca Raton, Florida, 161.

Weast, R.C. and Selby, S.M. (1966). Handbook of Chemistry and Physics, 47* ed; Ctiemical Rubber Co., Ohio.

Langer, H. (1947). Handbook of Physical Constant; Ohio Chem.

CHMT^'KIII 139

^ferences

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Q Dijkshoorn, W.; Van Broekhoven, L.W. and Lampe, J.E.M. (1979)./\gr/c. Sci., 27: 241.

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• Eigen, H. (1994). Ber Forsenungszent. Junelich. Juel-2920. 136.

G Hall, W.S.; Patoezka, J.B.; Minenda, R.J.; Poter, B.A. and Miller, E. (1980). Arch. Environ. Contam. Toxicol., 18: 765.

• Hirst, J.M.; Le Riche, H.H. and Bascomb, C.L. (1961). Plant Pathol., 10: 105.

• Hughes, M.K.; Lepp, N.W. and Phipps, D.A. (1980). Adv. Ecol. Res., 11: 217.

• Husain, M.Y. and Ahmad, F. (1993). Oxid. Commun., 16: 62.

• Jain, V.K. (1974). Fundamentals of Plants Physiology; S. Chand and Company (Pvt.) Ltd., New Delhi.

• Khan, S.U. and Khan, N.N. (1983). Plant and Soil, 74: 387.

• Khan, S.U.; Khan, N.N. and Nandan, D. (1983). J. Indian Soc. Soil Sci., 31: 325.

• Khan, S.U.; Khan, N.N. and Iqbal, N. (1985). J. Indian Soc. Soil Sci., 33: 779.

• Khan, S.U.; Qureshi, M.A. and Singh, J. (1996). Indian J. Environ. Health, 38: 1.

• Klingman, G.C. (1973). A Textbook of Weed Control as a Science; Wiley Eastern Pvt. Ltd., New Delhi, 81 .

CKM^<Sini 140

• Kowalezyk, Jh. (1993). Umweltkommum., 2"* 1292; 2: 365.

u Kozhevin, P.A.; Robinovich, N.L.; Belov, A.P. and Gradova.

H.B. {^99A). Pochvoved, 1:55.

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Qual., 5: 330.

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• Tyler, G. (1981). Soil Biochemistry; Marcel Dekker, New York, p. 372.

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CHAPTER W

EFFECT OF FLY ASH ON THE PHYSICO-

CHEMICAL PROPERTIES OF SOIL AND

SEED GERMINATION, GROWTH AND

METALS UPTAKE OF BARLEY AND

WHEAT PLANTS

chapter IV

INTRODUCTION

^ ^ - ly ash, released in bulk from thermal power stations and other

n f ^ industries, is a major waste product in India. The total fly ash

production in India is estimated to be 2-3 times more than that

in other countries (Gupta and Birendra, 1998), since most of the thermal

power stations and other industries are utilizing coal as a source of

energy on the basis of its availability in abundance. Finally, fly ash is

disposed off in the surrounding low-lying areas, waters and soils, resulting

in the huge environmental and ecological problems. Fly ash contains

appreciable amounts of plant nutrients (Plank and Martens, 1974;

Andojumpei et al., 1986; SIkka and Kansal, 1994; MIshra et al., 1995;). It

has the ability to increase the porosity of soil and thus to enhance the

permeability of water needed by the plants (Campbell et al., 1983). It can

also alter the pH of the soil (Hodgson et al., 1982; Khan et al., 1996).

However, sometimes it reduces the toxicity of trace metals by their

adsorption in excess amounts over fly ash surfaces. The use of fly ash is

found to increase the crop yields and is also capable of improving the

physico-chemical characteristics of soils (Chang et al., 1977; Elseewi et

CJd^Fm^i'p' 142

al., 1978; Page et al., 1979; Hill and Lamp, 1980; Khan et al., 1996;

Sarangi and Mishra, 1998).

In addition to this, fly ash also contains a wide range of harmful

metals like As, Cd, Pb, Sb, Ni, Cr, etc. (Majumdar and Mukharjee, 1983;

Thicke, 1988; Sikka and Kansal, 1994) along with other colloidal non-

metallic contaminants (Page et al., 1979) such as boron, etc. When, the

concentrations of cadmium and lead exceed their normal limits, they tend

to inhibit the growth of plants (Wong and Bradshaw, 1986; Oza and

Prasannakumar, 1996) and therefore, the excessive use of fly ash is

harmful to the soil microbes and hazardous for crops (Pichtel, 1990;

Pichtel and Hayes, 1990) as well as for humans and animals (Underwood,

1971; Schmitt et al., 1972). It is, therefore, imperative that the use of fly

ash in agriculture is done with utmost care in a scientific manner so as to

minimize the risks of environmental pollution, which adversely affects the

health of human beings and animals.

In the present study, pot experiments were designed to evaluate the

seed germination, growth and metals uptake of barley (Hordeum vulgare

L.) and wheat (Triticum aestivum L.) grown on fly ash amended soil with a

view to asses the role of fly ash in the production of these cereal crops.

CHyi<prE^iv 143

EXPERIMENTAL

The soil (0-30 cm) and fly ash, used in this investigation, were

collected respectively from Agricultural Farm of Aligarh Muslim University

and Thermal Power Station, Kasimpur, situated in Aligarh district (UP),

India. The physico-chemical properties of the soil and fly ash are given in

chapter I (table IV) and chapter II (table VIM), respectively.

Determination of available metals in soil and fly ash:

Available metals in soil and fly ash were determined as follows:


Systronlcs flame photometer.

Double beam atomic absorption spectrophotometer (GBC

902).

0.01 M Calcium chloride solution.

1N Ammonium acetate solution (pH 7.0).

0.1 M Triethanolamine solution.

Standard solutions of Na, K, Ca, Cr, Mn, Fe, Co, Ni, Cu, Zn,

Cd and Pb metals.

1 N Hydrochloric acid.

DTPA extracting solution: To prepare 1 liter DTPA extracting

solution, 13.1 ml reagent grade TEA, 1.967g DPTA (AR

Grade), 1.47 g of CaCI2 and HCI were added in distilled water

maintaining the pH of solution to 7.3.

CfOl<pm<lill/ 144

Procedure:

Available Na, Mg, K and Ca were determined in neutral normal

acetate extract of soil and fly ash. The extraction was carried out by

mechanical shaking followed by filtration. Available Na, K and Ca were

determined by using 'Systronics' flame photometer. The quantities of

calcium plus magnesium were measured by EDTA titration using

eriochrome black "T" indicator at pH 10 (Jackson, 1958). The amount of

magnesium was then calculated by subtracting the value of Ca from

calcium plus magnesium.

Available Cr, Mn, Fe, Co, Ni, Zn, Cd, and Pb were determined in

DPTA extract of soil and fly ash (Tandon, 1993). To obtain the metal

extract, 10 gm of air dried soil was shaken with 20 ml of DTPA extracting

solution for two hours. The suspension was then filtered through Whatman

filter paper no. 42. Available Cr, Mn, Fe, Co, Ni, Cu, Zn, Cd and Pb were

determined in this solution by using double beam atomic absorption

spectrophotometer (GBC 902). The results are recorded in table XV.

Determination of physico-chemical properties of fly ash amended soil:

Physico-chemical properties of fly ash amended soil were

determined by methods used for soil analysis in chapter I.The results are

given in table XVI.

Determination of physiological parameters of barley and wheat plants:

Pot experiment was conducted to evaluate the physiological

parameters of barley and wheat plants grown on fly ash amended soil. For

the pot experiment.

cwi(pm<iii'i^ 145

Table XV

Concentration of available metals in soil and fly ash

Metals

Na

Mg

K

Ca

Cr

Mn

Fe

Co

Ni

Cu

Zn

Cd

Pb

Cone.

Soil

250

1220

296.3

2860

Nil

4.95

6.21

2.8

Nil

2.9

0.58

Nil

Nil

of available metals ( ppm)

Fly ash

320

13500

360

12600

2.6

5.8

20

4.0

2.68

4.2

3.4

4.17

3.8

CWiFFEIill^ 146

Table XVI

Physico-chemical properties of fly ash amended soil

Doses of fly ash

(g kg'^ soil)

0

5

10

15

20

25

30

35

PH

8.50

8.45

8.38

8.30

8.20

8.10

7.99

7.94

E.G. (dS m- )

0.9000

0.9100

0.9150

0.9170

0.9216

0.9620

0.9742

0.9913

Mechanical Composition (%)

Sand

63.00

61.21

59.59

58.10

56.75

55.50

54.34

53.28

Silt

24.00

26.31

28.41

30.33

32.0

33.70

35.29

36.58

Clay

13.00

12.48

12.00

11.57

11.25

10.80

10.37

10.14

Organic matter (%)

0.410

0.419

0.427

0.435

0.442

0.448

0.454

0.459

Values are mean of three replicates.

CJ{yi<ppE^ii^ 147

the soil was dried, crusiied, sieved and was filled in earthenware pots (1

kg per pot) containing varying amounts of fly ash (0,5,10, 15,20,25,30 and

35 g kg-1 soil). The soil, in each pot, was then wetted with distilled water

to about 60% of its water holding capacity. Out of the certified seeds of

barley and wheat, procured from the Quarsi Government Agricultural

Farm, Aligarh, ten seeds were sown in each pot for every treatment. Each

set were taken in three replicates. The pots were kept at 35 ± 10C in a

green house, while maintaining moisture levels by adding distilled water

as and when necessary. After seed germination, the sprouts were equally

thinned and allowed to grow for a total period of 90 days. The

physiological parameters such as seed germination, number of leaves,

shoot and root length and fresh and dry shoot weights were examined.

The results are recorded in table XVII.

Determination of metals uptal(e of barley and wheat:

Metals uptake of barley and wheat were determined as follows:


Systronics flame photometer.

Double beam atomic absorption spectrophotometer (G-BC 902).

Cone, nitric acid.

Cone, perchloric acid.

Cone, hydrochloric acid.

EDTA solution.

Eriochrome black "T" indicator.

Buffer solution (pH 10).

Standard solutions of metals.

afii^PTr.'Rn' 148

'^LY ASH

Effect of fly ash on the growth of barley plant

Fig. 12

(^n^PTE^JT 149

Effect of fly on the growth of wheat plant

Fig. 13

SEED GERMINATION NO. OF LEAVES

' ' « 15 20 25 30 35

SHOOT LENGTH

' ^ 10 15 20 25 30 35

ROOT LENGTH

' ^ 15 20 25 30 35

FRESH SHOOT WEIGHT

' ^ 10 15 20 25 30 35

DRY SHOOT WEIGHT

' '' '' '' 25 30 35 0 . — ^ = ^ « ^

' ' 10 i5^nm7 Cone. Of fly ash (gkg-1 soil)

Effect of fly ash on the

barley plant ?S!.^?f""'"«tlon and growth Of

SEED GERMINATION

NO. OF LEAVES

' ' lo^T^rTir 35

SHOOT LENGTH

' ^ '' 20 25 30 35

FRESH SHOOT WEIGHT

0 5 i rn fTTi r^^

DRY SHOOT WEIGHT

25 30 35 ' ' « i5~inriri7

Cone. Of fly ash (gkg-1 soil)

Effect of fly ash on the

>«'heat plant seed germination and growth of

cHji(pm<ii I'U 152

Table XVII

Physiological parameters of barley and wheat grown on fly ash amended soil

Doses of

fly ash (g

kg' soil)

0

5

10

15

20

25

30

35

0

5

10

15

20

25

30

35

SG

(%)

60

70

80

80

90

90

100

80

60

70

80

80

90

80

80

80

NL

(per plant)

5.0

7.0

7.2

7.8

8.0

8.2

8.8

7.0

5.2

5.1

6.4

6.4

7.0

6.8

6.5

6.5

Parameters

SL

(cm/plant)

WHEAT

20.1

40.3

42.0

46.0

51.0

56.0

65.5

45.0

BARLEY

60.0

58.2

70.5

72.0

75.6

75.5

72.0

71.5

RL

(cm/plant)

12.2

14.8

18.0

21.2

24.2

26.8

29.1

18.5

12.0

12.1

17.2

26.2

28.2

28.1

28.0

27.3

SWf

(g/plant)

2.4

4.0

4.2

4.3

4.7

5.2

5.6

4.1

3.3

3.2

4.0

4.2

4.8

4.8

4.2

4.2

SWd

(g/plant)

0.9

1.5

1.5

1.6

1.7

1.9

2.0

1.5

1.2

1.1

1.4

1.5

1.9

1.9

1.6

1.6

SG = Seed germination, NL RL = Root length, SWf

= no. of leaves, SL = Fresh shoot weight, SWu

Shoot length, •• Dry shoot weight.


CWFPEILII^ 153

Procedure:

The plants were removed from the pots and washed with distilled

water. After the proper washing, these were dried and grounded for the

determination of metals. One-gram sample of each treatment was

digested with 10 ml of acid mixture containing HNO3: HCI04 (4:1). The

digested samples were heated on a hot plate till the brown fumes ceased

and the digested mass converted into a syrupy liquid with some white

fumes (Piper, 1966). The samples of the syrupy liquid were dissolved in 5

ml of cone. HCI and diluted with distilled water. To obtain the clear

solutions, these were filtered and then adjusted to a volume of 50 ml in

each case. The amounts of available Na, K and Ca were estimated from

these solutions by using "Systronics" flame photometer. The quantity of

calcium plus magnesium was determined by EDTA titration using

eriochrome black "T" indicator at pH 10 (Jackson, 1958). The amount of

magnesium was then calculated by subtracting the values of Ca from Ca

plus Mg. The concentration of available Cr, Mn, Fe, Co, Ni, Cu, Zn, Cd

and Pb were estimated by double beam atomic absorption

spectrophotometer (GBC 902,). Results are recorded in tables XVIII and

XIX.

CHMT^^I"^ 154

Table XVIII

Effect of fly ash on the metals uptake by barley

Metals

Na

Mg

K

Ca

Cr

Mn

Fe

Co

Ni

Cu

Zn

Cd

Pb

0

3800

910

8000

3500

Nil

40

35

0.04

Nil

3

10

Nil

Nil

5

Doses of fly ash (g kg' soil)

10 15 20

Concentration of metals u

4300

1600

13000

3800

0.26

48

60

0.15

0.55

5

18

0.22

0.28

4900

2100

14500

4300

0.60

65

75

0.23

0.69

7

24

0.89

0.37

5500

2500

16400

5100

0.92

78

89

0.28

1.25

10

28

1.98

0.50

6500

2900

17800

6000

1.14

84

104

0.32

1.89

13

28

2.43

0.60

25 30

ptake (ppm)

7000

3500

19000

7200

1.46

90

128

0.41

2.18

17

22

2.87

0.68

8500

4000

20500

8300

1.83

98

148

0.50

2.59

22

20

3.29

0.76

35

8900

3200

18100

9900

2.32

114

139

0.63

2.95

30

14

3.48

0.86


cHji<p<m<iiiv 155

Table XIX

Effect of aly ash on the metals uptake of wheat

Metals

Na

Mg

K

Ca

Cr

Mn

Fe

Co

Ni

Cu

Zn

Cd

Pb

0

3500

900

7500

1200

Nil

90

30

0.03

Nil

4

6

Nil

Nil

5

Doses of fly ash (g kg'^ soil)

10 15 20

Concentration of metals i

4700

1100

9900

1600

0.05

100

52

0.12

0.49

5

10

0.18

0.19

5600

1300

13000

2400

0.08

120

70

0.20

0.56

7

18

0.79

0.23

7000

2200

15800

3900

0.12

135

89

0.30

0.69

12

20

1.84

0.29

8600

3700

19500

5500

0.16

150

100

0.45

0.76

18

28

2.24

0.37

25 30

iptake (ppm)

9900

3500

19500

6000

0.21 •

150

118

0.45

0.88

26

24

2.16

0.48

11600

2600

17100

6500

0.27

158

142

0.50

1.00

38

18

2.76

0.58

35

13900

1800

15800

7000

0.38

168

130

0.60

1.17

49

12

3.01

0.69


CWPPElill/ 156


The results of available metals (vide table XV) showed that

considerable amounts of macro and micronutrient elements, viz. Mg, K,

Ca, Mn, Co, Fe and Zn were present in soil and fly ash. The levels of

these nutrients were higher in fly ash than the soil along with some toxic

metals such as Cr, Ni, Cd and Pb.

The physico-chemical properties of fly ash amended soil (vide table

XVI) showed a variation in pH, electrical conductivity, mechanical

composition (sand, silt and clay) and organic matter contents with the

increasing doses of fly ash as compared to the natural soil. The pH and

percentage of sand and clay contents of the soil were decreased by

addition of increasing doses of fly ash. The decrease in pH and

percentage of sand and clay contents may be attributed to the slightly

acidic nature of fly ash (pH 6.72) containing lesser quantities of bigger

and smaller size particles assigned to sand and clay fractions. On the

other hand, electrical conductivities and percentage of silt and organic

matter contents were significantly increased on increasing doses of fly

ash, which may be due to the increase of soluble and /or available salts

(Moliner and Street, 1982; Khandkar et al., 1999) and percentage of silt

particles and organic matters in treated soil.

The chemical analysis of plants (vide table XVIII and XIX) showed

that the concentration of metals uptake were higher in barley than in

wheat. The concentration of Mg, K, Fe and Zn were found to increase at

C7Ca<P2?E(^/1/' 157

lower doses of fly ash addition upto 30 and 20 g kg'"" soil in barley and

wheat, respectively and thereafter, it was found to decrease. On the other

hand, the uptake of Cr, Mn, Fe, Co, Ni, Cu, Cd, and Pb were increased on

increasing doses of fly ash throughout the entire range of study in both

case. The increased concentration of metals uptake by plants may be due

to the lowering of soil pH, capable of providing the greater amounts of

available metals to the plants. However, the increase of preferential toxic

metals uptake may be the cause of inhibition for nutrient metals such as

Mg, K, Fe and Zn (Chaudhary and Wallace, 1976; Watkins and Ferguson,

1982; Pandit and Prasannakumar 1999) in both plants. The toxic metals

were also found to inhibit the bioactivities in soil.

The effect of fly ash on the seed germination and growth of barley

and wheat plants (vide fig. 12-15 and table XVII) have indicated a

beneficial effect of fly ash up to 30 and 20 g kg" soil in cases of barley

and wheat, respectively and then a phytotoxic effect was noticed beyond

these doses. The beneficial effect of fly ash at lower doses could be

ascribed to the improved soil conditions such as porosity (Saxena et

al., 1998), availability of nutrients (Khan et al., 1996), organic matter

along with enhanced microbial activities, needed in the release of

nutrients for healthy plant growth. However, increased levels of toxic

metals at higher doses of fly ash imposed the phytotoxic effect

(Martensson, 1992; Oza and Prasannakumar, 1996) in creating the

abnormality in the absorption of the plant nutrients that has resulted in

their stunt growth.

C7C3<PZiE!R./1^ 158

^ferences

Q Andojumpei, N.J.; Uzawa, M.N. and Yoji (1986). Nippon Dajo

Hiryogaku Zasshi, 57:391.

• Campbell, D.J.; Fox, W.E.; Altken, R.L. and Bell, L.C. (1983).

Aust. J. Soil Res., 21:147.

• Chang, A.C.; Page, A.L. and Warneke, J.E. (1977). J. Environ.

Qual., 6:267.

• Chaudhary, F.M. and Wallace, A. (1976). Commun. Soil Sci. Plant

Anal., 7:89.

u Elseewi, A.A.; Bingham, F.T. and Page, A.L. (1978). J. Environ.

Qual., 7:69.

• Gupta, A. and Birendra, K. (1998). Res. J. Chem. Environ., 2:23.

• Hill, M.J. and Lamp, C.A. (1980). Aust. J. Exptl. Agric. Animal

Husb., 20:377.

• Hodgson, L.; Dyer, D. and Brown, D.A. (1982). J. Environ. Qual.,

11:93.

• Jackson, M.L. (1958). So/7 Chemical Analysis; Prentice Hall Inc.

Englewood Cliffs, N.J.

• Khan, S.U.; Begum, T. and Singh, J. (1996). Indian J. Environ.

HLTH, 8:41.

• Khandkar, U.R.; Gangwar, M.S. and Srivastava, P.C. (1999). Poll.

Res.; 18: 101.

• Majumdar, A. and Mukharjee, S.N. (1983). Fuel Sci. Technol., 2:

80.

CHM^'HI'^ 159

• Martensson, A.M. (1992). So/7 Biol. Biochem., 24:435.

u Mishra, A.K.; Williams, A.J.; Bhowmik, A.K. and Banarjee, S.K.

(1995). Soil and Plant Analysis; Hans Publisher, Bombay.

• Moliner, A.M. and Street, J.J. (1982). Soil Crop Sci. Soc; Florida

Proc. II: 217.

• Oza, R.A. and Prasannakumar, P.G. (1996). Phytotoxic effect of

Cd II on the growth of Phaseollus aureus. L.; Proc. 83'^^ Ind. Sci.

Cong. Abstract, p. 55.

• Page, A.L.; Elseewi, A.A. and Straughan, I.R. (1979). Residue.

Rev., 71:83.

• Pandit, B.R. and Prasannakumar, P.G. (1999). Poll. Res., 18:483.

• Pichtel, J.R. (1990). Environ. Pollut., 63:225.

• Pichtel, J.R. and Hayes, J.M. (1990). J. Environ. Qual., 19:593.

• • Piper, C.S. (1966). So/7 and Plant Analysis; Hans Publishers,

Bombay.

• Plank, C O . and Martens, D.C. (1974). So/7 Sci. Soc. Am. Proc,

38:974.

• Sarangi, P.K. and Mishra, P.C. (1998). Res. J. Chem. Environ., 2:7.

• Saxena, M.; Ashokan, P. and Chauhan, A. (1998). Clay Res.. 17: 109.

• Schmitt, N.; Philion, J.J.; Carsen, A.A.; Harnader, M. and Lynch, A.J. (1972). Can. Med. Ass. J., 121:1474.

a SIkka, R. and Kansal, B.D. (1994). Bioresour. Technol., 50:269.

• Tandon, H.L.S. (1993). Methods of Analysis of Soil, Plants, Waters and Fertilizers: F.D.C.O. (Pub.), New Delhi.

cmPM^lV 160

• Thicke, F.F. (1988). Effect of Scrubber Sludge on Fly Ash on Soil Properties and Crop Growth; Ph.D Thesis, Univ. of Illinois at Urbana Champaign.

• Underwood, E.J. (1971), Trace Elements in Human and Animal Nutrition, 3"^ ed.; Academic Press, New York.

• Watkins, C.B. and Ferguson, I.B. (1982). Sci. Hortic, 17:319.

• Wong, M.H. and Bradshaw, A.D. (1986). New Phytol., 91:255-61.

CHAFTEE ¥

EFFECT OF ENDOSULFAN ON THE SEED

GERMINATION, GROWTH AND

NUTRIENTS UPTAKE OF FENUGREEK

PLANT

ChajpterV

INTRODUCTION

C5y^ he demand for a variety of organic pesticides is increasing many

^^ folds day by day to maximize the crop production. Among the

pesticides, endosulfan is a non-ionic systemic, contact and

stomach insecticide. It is commonly used insecticide to prevent the

infestation of a variety of crops by harmful insects and mites. Endosulfan

is an organochlorine insecticide, which has long residual action and low

solubility in water. Thus, it is highly persistent and its prolonged use may

lead to its accumulations in animals and human beings through food chain

(Indraningsih, 1993; Nikolenko and Amirkhanov, 1993). Ignacimuthu and

Sarvana (1994) have reported that the endosulfan causes irreparable

chromosomal damage in Allium cepa L. An acute toxicity of this pesticide

has also been reported on Bombyx mori (Maruthanayagam et al., 1997) and

other organisms (Fromm and Filser, 1991). Sahai and Chaudhari (1995)

observed that endosulfan causes severe adverse effects on the blood

cells in rats. It also causes a genotoxic effect on aquatic animals (Gopal

et al., 1980; Reddy et al., 1991; Murugappan and Guna, 1996). Ahmad et

al. (1993) have reported that reproductive organs and sex hormones of

CWPT^^i^ 162

neonatal rats were damaged by endosulfan toxicity. It has been found to

be absorbed by plants as such and is phytotoxic for their growth (Kotadia

and Bhalani, 1992; Sabale and Misal, 1993; Sharma and Singh, 1993).

For obvious reasons, endosulfan is banned in some parts of the

world. Thus, it was thought useful to study the effect of endosulfan

treatment on seed germination, growth and nutrients uptake of fenugreek

{Trigonella foenumgraecum L.) plant under Indian climatic conditions

where its use still continuing.

cm'FrE'R.'U 11631

EXPERIMENTAL

The soil (0-30) used in this investigation, was collected from

Malhepur village, besides the Aligarh-Ramghat Road, situated in district

Aligarh (U.P.), India. The soil was air dried, crushed and sieved through

100 mesh sieve (BSS) and then physico-chemical properties were

determined by the methods used for soil analysis in chapter II and IV.

Results are given in table XX.

Determination of physiological parameters of fenu-greek plant:

For pot experiment, the soil was air dried, crushed and sieved and

then it was filled in earthenware pots (1 kg per pot) containing varying

quantities of endosulfan (0, 250, 500, 750 and 1000 mg kg-1 soil). The

soil, in each pot was then wetted with distilled water to about 60% of its

water holding capacity. Out of the certified seeds of fenugreek, procured

from the Quarsi Government Agricultural farm, Aligarh (U.P), India, ten

seeds were sown in each pot for every treatment. Each set was taken in

three replicates. The pots were kept at 35 ± 10C in a green house, while

maintaining moisture levels by adding distilled water, when necessary.

After seed germination, the sprouts were equally thinned and allowed to

grow for a total period of 90 days. The physiological parameters such as

seed germination, number of leaves, shoot and root length and fresh and

dry shoot weights were examined. The results are recorded in fig. 16 and

17 and table XXI.

Determination of nutrients uptake of fenugreek plant:

Metals uptake of fenugreek plant was determined by following

methods:

CKji'PPEIil' 164

Table XX

Physico-chemical properties of soil

Parameter

PH

Electrical conductivity (dS m" )


Sand

Silt

Clay

Organic matter (%)

Available nitrogen (mg kg'^ soil)

NH/-N

NOs'-N

NO2-N

Available nutrients (ppm)

Macronutrients

Mg

P

K

Ca

Micronutrients

Mn

Co

Fe

Cu

Zn

Values

7.2

0.4500

71.08

19.72

9.20

0.41

58.00

38.0

9.50

1640

45.2

396.0

3092

7.29

3.6

10.29

4.8

1.28

i.'if^-m^'KT. 165

1 = Control, 2 = 250 mg, 3= 500mg, 4 = 750 mg, 5 = lOOOmg

Effect of endosulfan on the growth of fenugreek plant

Fig. 16

166

SEED GERMINATION NO. OF LEAVES

750

SHOOT LENGTH ROOT LENGTH

FRESH SHOOT WEIGHT DRY SHOOT WEIGHT

750 1000

Cone, of endosulfan (mg kg'^ soil)

Effect of endosulfan on the seed germination and growth of fenugreek plant

Fig. 17

CKA'PPE'R.'^ 167

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CO T—

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P c\i CO

00 1 ^ CO

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CKji'FrEli'U 168


Systronics flame photometer.

Double beam atomic absorption spectrophotometer (GBC

902).

Cone, nitric acid.

Cone, perchloric acid.

Cone, hydrochloric acid.

EDTA solution.

Erioehrome black "T" indicator.

Buffer solution (pH 10).

Standard solutions of K, Ca, Mn, Fe, Co, Cu and Zn metals.

Vanadomolybdate solution.

Phosphorous standard solution (50 ppm).

Procedure:

Plants were removed from the pots and were properly washed with

distilled water, dried and grounded and were used for the determination of

nutrient elements. Out of these, one-gram sample of each treatment was

CKA'PM^V 169

digested with 10 ml of acid mixture containing HNO3: HCI04 (4:1). The

digested samples were heated on a hot plate till brown fumes ceased and

the sample got converted into a syrupy liquid along with some white fumes

(Piper, 1966). These were dissolved in 5 ml of cone. HCI and diluted with

distilled water. To obtain the clear solutions, these were filtered and then

adjusted to a volume of 50 ml with distilled water in each case. The

amounts of available K and Ca were measured in these solutions by using

"Systronics" flame photometer. The quantity of calcium plus magnesium

was determined by EDTA titration using eriochrome black "T" indicator at

pH 10 (Jackson, 1958). The amount of magnesium was then calculated by

subtracting the value of Ca from Ca plus Mg. The concentration of

available Mn, Fe, Co, Cu and Zn were estimated by double beam atomic

absorption spectrophotometer (GBC 902). The amount of P was estimated

by spectrophotometeric procedure at 470 nm by using vanadomolybdate

solution as colouring agent (Chopra, 1979). The results are recorded in

table XXII.

cW'Fmii'U 170

Table XXII

Effect of endosulfan on the nutrients uptake of

fenugreek plant

Nutrients

Mg

P

K

Ca

Mn

Fe

Co

Cu

Zn

Doses of endosulfan (mg kg' soil)

0

Cone

4500

4031

28000

13635

60.1

70.3

0.046

7.3

32.0

250

entration

4310

3530

24000

15400

75.3

90.2

0.159

10.4

39.0

500

of nutriei

3800

3000

20860

15520

90.1

120.4

0.270

14.2

43.2

750 1000

its uptake (ppm)

3123

23.06

16083

14000

80.3

135.1

0.190

12.0

53.4

2432

1608

12000

12630

60.4

165.0

0.140

9.9

65.9


CKA'PTEIL'^ [ 1 7 1 I


It is evident from the table XX that the soil, used in this

investigation, was nearly neutral {pH 7.2) in reaction, which is fairly good

for the survival of soil microorganisms responsible for the maintenance of

soil fertility. The soluble salt contents are low as revealed by low electrical

conductivity. The soil was sandy loam with low clay and organic matter

contents indicating a meager adsorptive power for the organic pesticide

applied on its surfaces. However, it contains a considerable amount of

available nutrient elements viz. Mg, P, K, Ca, Mn, Co, Cu and Zn. Hence,

the soil is fairly good to conduct the investigation on the role of organic

chemicals used in soil environment where clear changes could be

visualized without masking effect posed by the excess nutrients as

contained in a fertile soil. Thus, Aligarh soil has become a fascinating field

of soil research in recent years (Singhal and Bansal, 1978; Khan and

Singh, 1996; Khan et. ai, 1999; Khan etai, 2000).

The results on the chemical analysis (table XXII) of fenugreek plant

grown on endosulfan amended soil indicate that the concentration of Ca,

Mn, Co, and Cu uptake were found to increase at its lower doses (up to

500 mg kg""" soil) and thereafter an inhibitory trend in adsorption was

noticed. However, the concentration of Mg, P, K, Fe and Zn absorption

was restricted in presence of endosulfan even at its lower levels with

respect to control. The physiological observations (fig. 16 and 17 and

table XXI) of fenugreek plant indicate that the natural soil could very well

CHyKFfE'R^'U 172

support the germination and growth of this plant. However, the application

of varying quantities of endosulfan in a series of pot experiments revealed

that it is phytotoxic even at lower doses with regards to seed germination,

growth and nutrients uptake. The effect becomes more severe with

increasing levels of the insecticide. The inhibitory effect on seed

germination, growth and nutrients uptake of fenugreek might be due to the

toxic effect of endosulfan on Azetobactor spp., Rhizobium spp.,

Cellulolytic microorganisms, phosphate dissolving microbes and other soil

microorganisms (Smith, 1945; Chiaro, 1953; Mahmoud et al., 1970;

Parr, 1974; Khan et al., 1987), which determine the soil fertility. It may also

be due to the inhibition in number of nodules, lateral roots in nodulating

legumes and mycorhizal symbioses that can alter the growth and activity

of nitrifying bacteria viz. Nitrosomonas and N/trobacter. The possibility of

chromosomal damage (Ignacimuthu and Sarvana, 1994) posed at higher

levels of endosulfan absorption in fenugreek plant grown in soil of low

adsorptive capacity (Tiwari et al., 1994) cannot be ruled out.

Cm'PTEIi'i^ 173

9fiferences

a Ahmad, M.M.; Ahmad, M.M. and Sarvat, S. (1993). Pak. J. Zoll.,

25: 11.

• Chiaro, G.D. (1953). Agr. Ital., 8: 343.

• Chopra, S.L. (1979). Studies in Leactiing and Adsorption of

Pesticides in soils and the effect of their Residues on Plants, Final

report of PL-480 Project (A7-PSR 5), p. 102.

• Fromm, H. and Filser, J. (1991). Verh. Ges. Oekol., 20: 61.

Q Gopal, K.; Anand, M.; Khanna, R.M. and Misra, D. (1980).

Toxicol, Lett., 7: 451.

• Ignacimuthu, S. and Sarvan, K.P. (1994). J. Ecetoxico. Environ.

Monit., 4: 211.

u Indraningsih; McSweeny, C.S. and Ladds, P.W. (1993). Aust. Vet.

J., 70: 59.

• Jackson, M.L. (1958). So/7 Chemical Analysis; Prentice Hall Inc.

Englewood Cliffs, N.J.

• Kotadia, V.S. and BhalanI, P.A. (1992). Gujrat Agric. Univ. Res. J.,

17: 161.

• Khan, S.U, and Singh, J. (1996). Indian J. Environ. HLTH, 38: 153.

• Khan, S.U.; Shafiullah and Khan, Jamal A. (1999). Indian J.

Environ. HLTH,4^^. 83.

• Khan, S.U.; Khan, Jamal A.; Bhardwaj, R.K. and Jabin, S.

(2000). J. Indian Chem. Soc, 77: 326.

• Khan, S.U.; Khan, N.N. and Khan, F.A. (1987). Environ. Pollut.,

47:115.

CWPT^'K.1^ 174

a Mahmoud, S.A.Z.; Salim, K.G. and Elmokalem. M.T. (1970).

Zentralbl. Bakterlal Abt II., 125:134.

u Maruthanuyagam, P.; Krishnamothy, P. and Kannan, S. (1997).

Pollut. Res., 16: 271.

u Murugappan, R.M. and Guna, S.M. (1996). J. Ecotxico Mont., 6:

131.

• Nikolenko, A.G. and Amirkhanov, D.V. (1993). Agrokhimiya, 4:

115.

• Parr, J.C. (1974). Effect of Pesticides on Microorganism in Soil and

Water; Soil Sci. Am., Madison, USA, p. 315.

• Piper, C.S. (1966). So/7 and Plant Analysis; Hans Publisliers,

Bombay.

• Reddy, A.N.; Venugopal, N.B.R.K. and Reddy, S.L.N. (1991).

Pestic. Biochem. Physiol., 39: 121.

• Sabale, A. and Misal, B. (1993). Geoblo., 20: 166.

• Sahai, S. and Chaudhary, S. (1995). Proc. Acad. Environ. Bio., 4:

193.

• Sharma, D.C. and Singh, IVI. (1993). Indian J. Agr. Sci., 63: 59.

• Singhal, J.P. and Bansal, V. (1978). Soil Sci., 126: 360.

• Smith, N.R.; Dawson, V.T. and Wenzel, IVI.E. (1945). So/7 Sci. Soc.

Am. Proc, 10: 197.

• Tiwari, S .C; Singh, G.R. and Chattoraj, A.N. (1994). Interaction

of Insecticides with Plant Systems; Soil Environment and

Pesticides, (Editoprs: Prasad, D. and Gaur, H.S.) Venus Publishing

House, New Delhi, p. 137.

INDIAN J. ENVIRON HLTH. Vol 41. No 2, P. 83-89 (1999)

Mobility of Amino Acids through Soils amended with some Pesticides

S. U. KHAN*. SHAFIULLAH** AND JAMAL A. KHAN**

The mobility of amino acids was measured in terms of R, values by using soil-thinlayer chromatography which shows the mobility order : valine > alanine > phenylalanine > glycine > lysine in natural soil. The amendment of soils carried out by pesticides viz. paiathion-methyl, dimethoate, carbendazim, chlorpyrifos, endosulfan and methyl-demeton have shown variable degree of influence on the mobility of amino acids. The mobility of valine, alanine and lysine was found to decrease whereas, that of phenylalanine was found to increase on increasing concentration of pesticides throughout the range of their amendments (upto 100 mg 100 g' soil). However, the enhanced mobility for glycine in case of parathion-methyl and dimenthoate upto 75 mg and carbendazim, chlorpyrifos, endosulfan and methyl-demeton upto SO mg 100 g' soil was observed and thereafter it declined. The results have been explained on the basis of the nature and chemical behaviour of pesticides and amino acids in soil medium (pH = 8.5).

Key words: Amino acids. Pesticides, Soil-thinlayer Chromatography.

Introduction

The significance of amino acids and pesticides are well known in relation to soils and plants. Amino acids are found to exist in soils as a result of the decomposition of proteins and other soil organic matters'-^ by soil micro-organisms and play a vital role in biochemical processes^. They form a variety of complexes* with clays, lignins and other soil organics and may undetgo polymerisation reaction. The products such as indoles act as growth promoting substances, hormones or auxins in plants'. However, when pesticides reach in soil either by direct application or run

off from treated crop plants, get adsorbed onto soil colloids, which have been intensively studied*'' in recent years. It is found that pesticide treatment led to both qualitative and quantitative changes in the free amino acids composition of soils". Some studies have also been made to investigate the mobility of amino acids' and pesticides'" in soil and pesticides effect on soil microbial population growth"'".

In view of the significant relationship attached to amino acids, soils and pesticides the present study was conducted to investigate the mechanisms involved in the mobilization processes of certain amino acids through pesticide amended soils.

•Department of Applied Chemistry, Z.H. College of Engg. and Tech., Aligarh Muslim University, Aligarh-202002 (India)

••Department of ChemisUy, Aiigarh Muslim University, Aligarh-202002 (India)

83

84 KHAN ET EL

Materials and Methods

The study was conducted on the illitic fine sandy loam collected from Agriculture farm of Aligarh Muslim University, situated in Aligarh District, India. The physico-chemical properties of soil were : Sand, 63%; Silt, 24% and Clay, 13%, pH, 8.5 (1:2.5 soil water ratio); CEC, 16.3 m eq 100 g 'soil; Organic matter, 0.41%; Exchangeable Na*, 1.0; K*, 0.5; Ca2% 3.5 and Mg^*, 1.2 meq 100 g ' soil; Available NH%-N, 32.0;NO^-N,6.0; NO3-N,40.0; P, 25.0 and K. 76.0 m eq kg soil.

The natural soil was amended with varying quantities of pesticides viz. parathion-methyl, dimethoate, carbendazim, chlorpyrifos, endosulfan and methyl-demeton containing O, 25, 50, 75 and 100 mg 100 g ' soil. A soil slurry of these amended soils was prepared in distilled water with a soil water ratio of 1:2 in each case and was applied on glass plates (20x5cm) with the help of an appplicator in order to get an uniform layer of 0.5 mm thickness. The plates were then air dried at room temp. 30 ± 3°C. Two lines were scribed at 4 and 14 cm from the base at the surface of all plates. An aqueous solution of 10 1 amino acids (0.05 M) was spotted at the base line with the help of micropipette. These spotted plates were developed in closed-chamber using distilled water as the mobile phase to a distance of 10 cm. The plates were then air dried. The amino acids were detected by spraying a solution of 0.2% ninhydrin in ethanol (w/v), heated in an oven at 70-80°C for 10-15 minutes that gave a pink

coloured spot. The mobility in terms of R, values was calculated by the formula,

Rf = Distance moved by amino acid

10

Results and Discussion

The mobility of amino acids (Fig. 1) in natural (O dose) soil follows the order : valine > alanine > phenyl alanine > glycine > lysine. These trends are in accordance with the increasing order of their molecular weights except in case of phenylalanine and lysine. The lower mobility of phenylalanine as compared to valine and alanine, may be due to its lower solubility (Table 1) in water. On the other hand, the lowest mobility of lysine may be due to its higher adsorptive tendency over soil colloids in which the chemical structure plays a decisive role in its chemical behaviour as -

H3N-C-H

COO

Soil pH=8.5. H,N-C-H

hi.

Lysine (neutral Lysine (basic medium) medium)

Thus, due to the presence of two protonated amino groups in neutral medium out of which, one still remains in the cationic form even in basic medium. Thus, the molecule behaves as more adsorptive over negative charged sites of the soil colloids with strong electrostatic force resulting in its lowest mobility. Its adsorption

MOBILITY OF AMINO ACIDS 85

on soil colloids can be illustrated as-

Soil colloids + H 3 N - (

yH, (CH,)-C-COO->

IjJH,

col ids>NH3-(CH,V(f-COO

H

Ihble 1. Structures and Characteristics of Amino Acids

Amino Acids Mol. weight Structures" Solubility'* (in Nature" gm/100 ml water)

Lysine

Glycine

Alanine

Valine

146.19

75.07

Phenylalanine 165.00

89.10

117.00

cpo-

HjN—C—H

NH, cpo-H,N—C—H

A

H3N—C—H

.1 ,00-

H3N—C—H

CH3

^cpo-

H3N—C—H

CH, CM,

24.99

2.96

16.51

8.85

Positively charged

Polar but uncharged




86 KHAN ET EL

The mobility order: valine > alanine > glycine can be explained on the basis of their increasing polarity order. The structures of valine, alanine and glycine (Table 1) contain secondary, primary and no alkyl groups respectively at their one end of the molecule, capable of inducing electrons (+1 effect) towards the carbon attached with -^NHj and COO" groups in these amino acids. Thus, this part becomes rich in electron density but, poor in +ve charged centre with diminishing cationic tendency in the increasing order of the number of methyl groups attached in these amino acids. The decreased cationic and increased anionic tendency has compelled these molecules to follow the above order of mobility in soil against the negative charged sites over soil colloids.

The effect of pesticides in soil on the mobility of amino acids (Fig. 1) shows a decreasing trend in case of lysine, alanine and valine in the order: parathion-methyl > dimethoate > carbendazim > chlorpyrifos > endosulfan > methyldemeton. The increased mobility of phenylalanine was found in the reverse order throughout the entire range of pesticides (upto 1(X) mg 1(X) g ' soil). On the otherhand, the mobility of glycine increased at lower concentration and thereafter it declined at higher concentration of pesticides addition. The varying mobility trends of amino acids in various pesticides systems can be understood by considering the chemical behaviour as supported by their structure.

CH,0 ' 0

-3°>to^H: H O ^ ctiu

CH3-N-C-CH2-S-Pr^ "*

OCH3

Parathion-aethyl Dlmethoat*

O OCH^

CI

Carbttndazia Chlorpyrif o«

CH3-CH2-S-CH2-CH2-S-PC ii/CCHj

OCH3

Methyl-daaaton


1.0^ M e t h y l - , Q T \ p a r a t h i o n ^ » t \ Dimethoate,. TN^arbendazi

100

Concentration of Pesticides (ag 100 g soi l )

Fig. 1 Mobility of Amino Acids through Soils Amended with Pesticides

88 KHAN ET EL

The maximum decrease in mobility of lysine, alanine and valine in parathion-methy 1-soil system may be due to the presence of nitro (-NO,) group which is capa-

Soll-Ptfathien- Mlfchyl

ble of formation of strong hydrogen bonding with amino or protonated amino groups in these amino acids at soil pH 8.5 as, follows -

H '

I ' I

Itost part of ••Ino acid

the

Hydrogen bonding

SoiloParathlon-nat^yl

The capacity for the formation of hydrogen bonding, is governed by the number of electronegative nitrogen or oxygen atoms in these pesticide molecules, that are capable of retaining these amino acids seems to be in the order: parathion-methyl > dimethoate > carbendazim > chlorpyrifos > endosulfan > methyl-demeton. Thus, the mobility of amino acids have been found to decrease in all these systems with increased doses of pesticides. However, the case of phenylalanine seems to be governed by its incapability for the formation of hydrogen bonding at increased concentration of pesticides, may be due to the presence of bulky phenyl group, that pose a steric hindrance which has resulted in increase of its mobility. On the otherhand, the polarity of glycine increases its mobility at lower concentrations (parathionmethyl and dimethoate upto 50 mg and carbendazim, chlorpyrifos, endosulfan and methyl-demeton upto 75 mg 100 g ' soil) due to the incapability of formation of hydrogen bonding but, at

H I

>l^—N'^ Raat part of tho I ; aaino acid H •

higher pesticides concentration the increased number of electronegative oxygen and nitrogen atoms of pesticides in soils systems are capable of diminish the mobility through hydrogen bonding. Rererences

1. Waksman, S. A. and Starkey, R.L. The decomposition of proteins by micro-organisms with particular reference to purified vegetable protein. J. Bact.. 23 : 405-428 (1932).

2. Waksman, S.A. and Lomanitz, S. Contribution to the chemistry of decomposition of proteins and amino acids by various groups of micro-organisms. J. Agr Research, 30: 263-281 (1924).

3. Theng, B.K.G. The Chemistry of clay organic reaction. Ch. IV Adam-Hilger London, pp. 159(1974).

4. Talibuddin, O. Complex formation between montmorillonite clays and amino acids and proteins. Trans, Faradaxs Sac, 51 : 582-590 (1955).

5. Waksman, S.A. Transformation of Nitrogen in soil: Nitrate formation and nitrate reduction. Soil Microbiology VII John Wiley and Sons Inc., pp. 168(1952).

6. Camazano, M.S. and Martin, M.J.S. Interaction of some organophosphorous ftesticides with vermiculite. Applied Clay Res. 2 : 155-165(1987).


7. Yanron, S. Some aspects of surface interactions of clay with organophosphorous pesticides. SoilSci., 125(4): 210-216 (1978).

8. Wainwright, M, and Pugh. G.J.F. Change in the free amino acid content of soil following fungicide treatment. Ibid.. 7 :1-4 (1975).

9. Singhal, J.P., Khan, S.U. and Bansal, V. Studies on the mobility of amino acids in soils by soil-thinlayer chromatography. Proc. Indian Nam. Sci. Acad.. 44(5): 267-270 (1978).

10. Khan, S.U. and Khan, N.N. The mobility of some organophosphorous pesticides in soils as affected by some soil parameters. Soil Sci. 142:214-222(1986).

11. Alexander, M. Introduction to soil microbiology, 2nd Edition New Yrok : Wiley and Sons (1977).

12. Greeves, M.P. Davies, H.A., Marsh. J.A.P and Wingfield, G.J. CRC Cril. Rev. Microbiol, 5 : 1-38(1976).

13. Martin, J.P. Influence of pesticide residues on soil microbiological and chemical properties. Residue Rev.. 4 : 96-129 (1964).

14. Parr, J.F. In : Pesticides in soil and water (ed. W.P. Guenzi). Effect of pesticides on microorganisms in soil and water. Soil Sci. Soc. Am. Madison U.S.A. pp. 315-340 (1974).

15. Lehninger, L. A. Principle of Biochemistry. The Jonha Hopkins University School of Medicine (1982).

16. Gross, D. and Grodsky, G. In sublimation of Amino Acids and Peptides. J. Am. Chem. Soc. 77; 1878(1955).

Poll Res. 18 (4): 511-514 (1999) Copyright © Enviromedia

INFLUENCE OF Ni (II) AND Cr (III)-HUMIC ACID (HA) COMPLEXES ON THE AVAILABLE MAJOR NUTRIENTS STATUS

(NPK) OF THE SOIL

SAMIULLAH KHAN*, JAMAL A. KHAN**, R.K. BHARDWAJ* AND JAIBIR SINGH*

'Department of Applied Chcmistiy, Z.H. CoUe\;c ofEn^g. and Tech. Aligarh Muslim University, Aligarh - 202 002; **Department of Chemistni, A.M.U. Ali^mh - 202 002

Key words : Nickel, Chromium, Humic acid. Plant nutrients.

ABSTRACT

The application of varying doses (0-100 ppin) of Ni-HA and (0-600 ppm) of Cr-HA complexes were found to influence the availability of majc r nutrients (NPK) /C, NH^ - N, NO^-N and NO;;-N; P and K in soils. The availability of NH'- N and NO",-\ was found to increase with the increasing doses upto 50 ppm for Ni-HA and upto 300 ppm for Cr-H.A complexes with respect to control and attained to their maximum levels in 21/28 days beyond which they tend to decline. However, the amount of available NO>N and K showed an increasing trend throughout the entire range of their study. The toxic effect on the availability of these nutrients by metal humates followed the order Ni-HA>Cr-HA. The results ha\e been explained on the basis of changes in chemical and microbial acti\'ities in soils.

INTRODUCTION

Application of sewage sludge, industrial wastes and waste water to the agricultural soils have generated pollutional problems as they contain many toxic heavy metals. Heavy metals have gained considerable importance because of their active participation in many metabolic reactions as well as their inhibitory role in the nutrient utilization by plants (Khan and Khan, 1^83). Humic acid, a fraction of soil organic matter (SOM), forms stable complexes with heavy metals, by virtue of the presence of many active sites in\'ol\'ing N, S and O atoms. The presence of these complexed metals have also revealed to show a stimulating effect on the microbial actixities which in turn, affect the nutrients availability in soil (Zibilske and Wagner, 1982).

Tyler et al. (1974) have reported that the presence of heavy metals in soil rich in organic

matter has resulted in stimulating tendencies in the process of nitrification as well as in the availability of P and K in soil. Hpwever, the specific role played by Ni-and Cr-HA complexes in altering the soil nutrient status is lacking in literature. Therefore, it was considered worthwhile to investigate the influence of these metal-humates e.g., Ni-and Cr-HA complexes on the availabilitv of major plant nutrients (NPK) in soils with a view to assess their role in creating pollutional problems, in regards to soil fertility and plant nutrition.

MATERIAL AND METHODS

Soil used in these investigations was Hirapur fine sandy loam, collected from the cultivated lands of Aligarh district (U. P.) from a depth of 0-30 cm. The soil sample was dried crushed and sieved. The physico-chemical characteristics of the soil were : sand, 61% Silt, 25% and Clay, 14% (Piper 1950), pH

512 KHANET.4L

8.8 (1:2.5 soil: water ratio); Organic matter, 0.41% (Walkley and Black, 1947); CEC, 7.3 meq 100 gm'; Exchangeable Na\ 1.0; K*, 0.5; Ca=*, 3.5 and Mg-'*, 1.2 meq 100 gm' soil (Jackson, 1958), Available NH;-N, 52.0; NO'-N, 6.0; NO3-N, 40.0 (Hesse, 1971), P, 25.0 (Olsen 1954) and K, 76.0 mg Kg"' soU.

The soil organic matter (SOM) was extracted by treating the soil with 0.1 N NaOH + 0.5N NajCO, mixture at 80°C (Dasai and Ganguly, 1979). The SOM was then fractionated in humic acid (HA) and hilvic acid (F. A.). The HA fraction was purified by using the technique of electrodialysis. The Ni-and Cr-HA complexes were prepared by treating with their respective chloride salt (O.IM) solution with 1% humic acid. (McBride, 1978). The crystals of these complexes were obtained by keeping the mixture over night. They were filtered, washed with double distilled water, oven dried at 25 ± 1°C and kept in a closed bottles for the experimental work.

The experiments were conducted in earthen pots (15 X 15 cm) which were cleaned and coated with coaltar to prevent the absorption of plant nutrients. One kg of respective amended soil having varying levels of 0.0 (Control), 25, 50, 75 and 100 ppm in case of Ni-HA and 0.0 (Control), 75, 150, 300 and 600 ppm for Cr-HA complexes, were taken in each pot with three replications. The requisite amounts of the Ni-HA and Cr-HA complexes were thoroughly mixed into the soil. The experiment was carried out under similar conditions of temperature at 25 ± 1°C and maintaining soil moisture level up to 60% of their water holding capacity. The soil samples were drawn up periodically by a soil sampler at a regular interval of 7 days and analysed for the available major nutrients (NPK).


Fig. 1 show that the varying doses of Ni-HA and Cr-HA and their incubation period had a marked influence on the availability of major nutrients (NPK) in soil. At lower concentrations, upto 50 ppm in case of Ni-HA and upto 300 ppm of Cr-HA, the availability of NH*-N and NO-j- N is found to increase beyond which they tend to

decline with respect to control and attained a maximum level upto a period of 21/28 days as shown in Fig. 1 (a/a; b/b). Their availabihty in regards to their respective concentrations followed the order : 50 >25 >0 (control) >75 >100 ppm for Ni-HA and 300 >150 >75 >0 (Control) >600 ppm for Cr-HA complexes indicating their tolerance limits upto 50 and 300 ppm for maintaining soil fertility status. Thus, the toxic effect of these metal humates in soil medium followed the order : Ni-HA > Cr-HA which are in the general agreement of earlier workers (Leaper, 1978). The increased availability of NH*^ - N and NO; - N upto these non toxic levels may be regarded due to the greater involvement of soil microorgarusms responsible in decomposing and converting non available-N from the nitrogen containing organic compounds into inorganic NH^ as the main product as shown by:

RCH (NH,) GOGH +Hp — Amino acid RCONH^ + Hp > NH, + CO, Amide

» RCHj OH+NH,+COj

2NH,+ CO,+ H p - •^(NH,)XO, Ammonium carbonate

The increased availability of NH*-N and their simultaneous gradual oxidation into NO; -N with the active involvement of nitrifying bacteria such as Nitrosomonas spp. respor\sible in its conversion process can not be ruled out. The decrease in concentration of NHt-N and NOr-N could be

4 i

attributed to the depletion of their accumulated concentration by the ei\hancement in its availability throughout the entire range of study upto 7 weeks. Fig. 1 (c/c') eind are shown by

Nitrosomonas spp. NH; + 2[0] > N O ; + H* + Energy.

Nitrobacter spp. N O J + [O] > N O ; + Energy.

Fig. 1 (d/d') indicate a significant increase in the availability of phosphorus (P) with the increasing concentrations upto 50 ppm and 300 ppm for Ni-HA and Cr-HA concentration beyond which they tend to decline with respect to control and attained

INFLUENCE OF Ni (II) AND Cr {I11)-HUM1C ACID (HA) 513

the maximum at 28/35 days. The higher availability with applied 50 ppm and 300 ppm may have been due to increased microbial activities and becomes toxic at their higher concentrations. With the passage of time, the increased P availability may be considered due to the two fold effects of microbial and chemical activities. The role of ascomycetes, actinomycetes and reintroduced fungus flora, mycorrhizal fungi etc., responsible for the decomposition of organic matter and capable in releasing the available phosphorus along with CO^ and HjS gas (Gray and Williams, 1975). The solubilizing effect of inorganic non-available phosphorus in to its available form is seemed to be the plausible explanation for its increase as indicated by :

-* 2H,PO/2Cfl (HCO,),

•^ 2H,PO,+3CaSO^

Ca3(PO )j + 6CO, + 6Hp—

Ca,(POJj + 3H2S + 60j-

The decrease availability at their higher doses was possibly due to the bacterial/chemical reduction of its available form to unavailable phosphene (Singhal et al., 1979).

H3POr 2H

-H,0 *H,P03

2H

-H,0 •^ H,PO,

4H

-2H,0 -^PH,

Fig 1. (e/e') shows a steady increasing trend in the available potassium (K) throughout the entire range of study. It may be possibly due to its release from the alumino silicate minerals caused by the solubilizing effects of organic acids produced during the biodegradation of SOM coupled in their

9

3 2

«

0

0 > <

cr . Humic ocicJ ;ompJ«x • O f ' o ' l

.'•>

10

49

to

JL.L. •A. L..,;.

0 >V I I I I i,

• ^ u H M 4 7 i)

<'j,- \ to

TO

•J 1 0 4 ' ' • IS <«

I I I so >t «• « T I I I I t t

^•§1 Eftec» o» Mr-; C f -Humfcocfd compUxcs on th t ovoilotoiiay of

fvH4*-N. N 0 ^ - \ , N 0 3 - N , P o n j K in soi l .

( N i - H A , c o m p l y - , . 0 0, ^ r 5 . - « . 5 0 . - , .75 . * 100 ppr, )

f Cr-H-A c o m p l e x . - 0 0, • * - 75 , . * - . l W . - — 3 0 0 . - # - 6 0 0 ppw '

514 KHAN £T/^L.

release from the exchangeable sites by other cations and/or H* is the possible explanation for this increase.

ACKNOWLEDGEMENTS

The authors are grateful to the CSIR for pro\'iding financial support for this work.

REFERENCES

Dasai. M.V.M, and Ganguly, A.K. 1979. Exchangeand non-

exchanges behaviour of cation species in the

interaction of metal ions with humixc and fulvic acids./

Mian Chemical Soc. 56 : 1116-1172.

Gray. T.R.G. and Williams, S.T. 1975. Soil micro-organisms

{1st edn.), London, Longman Group Ltd.

Hesse, P.R. 1971. A textbook of Soil Chemical Analysis, 200-2

London, John Murray (Publishers) Ltd.

Jackson, M.L. 1958. Soil Chemical Analysis, Prentice Hall,

Englewood Cliff. N. J.

Khan, S.U. and Khan N.N. 1983. Influence of lead nd cadmium

on growth and nutrient concentration of tomato

(Lycopersicum escu/entum) and egg plant (So/onum

melongena). Plant and Soil. 74 : 387-394.

Leeper. G.W. 1978. Managing the heavy metals on the land.

Dekker, New York.

McBride, M.B. 1978. Transition metal bonding in humic acid.

Soil So. 126:200-209.

Olsen, S.R., Cole. C.V.. Watanabe, F.S. and Dean, LA. 1954.

Estimation of available phosphorus in soils by

extraction with sodium bicarbonate. U.S. Department

Agri. Ore. No. 939 : 99.

Piper, C.S. 1950. Soil and Plant Analysis, University of

Adelaida Australia.

Singhal, J.P., Khan, S. and Nandan. D. 1979. Influence of cetyl

trimethyl ammonium bromide on the availability of

major nutrients from a fertilized soil and the response

of tomato and mustard. Indian j. Agric. Scl. 49 ; 423-426.

Tyler, G.. Mornsjo. B. and Nilsson, 8. 1974. Effects of

cadmium, lead and sodium salts on nitrification in a

mull soil. Plant and Soil. 40 : 237-242.

Walkley, A. and Black, C.A. 1947, A. Critical examination of

rapid method for determining organic carbon in soil.

Soil Sa. 63 : 251 -264.

Zibilske, L. M. and Wagner, G. H. 1982. Bacterial growth and

fungal genera distribution in soil amended with sewage

sludge containing Cd, Cr and Cu. Soil So. 134 : (6) 364-

370.

H i.> : K M I I 4 \ \ I I

J. Indiun Clu'm. Soc.. \ 111. 77. .)iil\ 2(HK). pp. Mk-na

Effect of some organic compounds on the mobility of some trace metals through soil amended with fly ash

Samiullati Khan". Janial A. Khan , R. K. Bhardwaj" and Shagufta Jabin"

"Deparimenl ot Applied Chemistry. Z. H. College of Engineering &. Technology, Aligarh Mushum University. Aiigarh-2()2 (K)2. India

''Departnient ot Chemistr>. Aligarh Mushurri University, AIigarh-202 002. India

Minui^i iii'i icicivid H.lur.i /VyV rfiised 21 February 2000. actepied J March -'••

The t-lliTts of some organic compounds, viz. formir acid, acetic acid, methanol, rthanol. fonnaldch>dc. acetaldehyde, acetone and i-th>l meth>l ketone on the mobility of some trace metals, viz. Co, Zn. t"u, \f. and Pb through soil amended with fly ash hJ • been intcsticalrd in an alkaline fmc sandv loam soil of .Aligarh district lU.P.i. The results indicate decrease in the mohilit) of trace metals with increasing dosapc of n> ash in soil but with a variation in intensity by use of organic compound as a mohik phase. The mobility order is Co > Zn > Cu > .Ag > Pb in case of organic acid* and alcohols, and Co > Cu > Zn > Ag > Ph for aldehydes and ketones. The effect of organic chemicals on the translocation of metals follows the sequence : formic acid > acetic acid > methanol > acetone > formaldehyde > ethyl methyl ketone > acetaldehyde > ethanol.

Various synthetic, volatile and non-volatile organic chemicals are released in the environment from the industries and chemical laboratories in appreciable quantities. It has also been revealed in literature thai several organic compounds such as aldehydes, ketones, carboxylic acids, etc, are produced in soil as a result of chemical' or biochemical-decomposition of organic matter used in agriculture and of the crop residues. The> are tound to play an important role in the translocation of plant nutrients as well as toxic metals and their avaiiabilii>' to plants. Higher dosages of these organic chemicals sometimes become fatal to plants and adversly affect the life processes of soil microorganism.

Among trace metals Zn. Cu and Co are regarded as micronutrients and are required for the healthy growth of plants and animals. Zn" acts as a prosthetic group of an enzyme in the con\ersion of tryptophan into indole acetic acid, the growth promoting horomone. Copper is required in certain biological oxidation-reduction systems, whereas cobalt is regarded to be essential for symbiotic N; fixation by some microorganism and in the synthesis of vitamin Bp needed for animal health. Silver and lead"* are hazardous for plants, microflora and soil microorganism^.

The use of fly ash as a source of plant niitrients tried in agriculture has become topic of research*. However, scanty information is available in literature on the role of fly ash in the translocation of trace metals through soil amended w ith it in relation of these soil organics. The present study was designed with a \ieu to ascertain the role of certain organic compounds, namely, acetic acid, formic acid.

. 26

ethanol. methanol, acetaldehyde, formaldehyde, acetone and ethyl methyl ketone on the translocation of some trace metals such as Co. Zn. Cu. Ag, and Pb through >.oil amended with fly ash.

Results and Discussion

li is evident from Fig. 1 that mobility of trace me'.jl^. nameK. Co. Zn. Cu. Ag and Pb varies with varying concentration of n\ ash in soil as well as with the nature o! organic compounds used as mobile phase. The mobili!> of various metal ions follow s the order : Co > Zn > Cu > .Ag > Pb, through soil amended with tly ash as a static phase and water or gi\en organic acids and alcohols used as mobile phase. These results are in close agreements with the uork of Gaszczyk ei al?. This order of mobility seems to be in-versel) related to the order of their binding capacit> ot given ions with the soil colloids^. The mobility order til metal ion^ in case of aldehydes and ketones follow-, the order ; Co > Cu > Zn > Ac Pb. which is slightly difierem from the above. The variation in the order of mobilities between copper and zinc in the s\ stems of acids and alcohols on one hand and in aldehydes and ketones on the other ma'* be attributed to the difference in the rate of their permeability and nature of functional groups present in the mobile phases of these systems. The mobility is found to decrease with the increasing dosages of fly ash in soil and is airributed to the enhancement in the adsorbing capacity caused by the unburnt carbon content (0.6%) in fly ash".

The order of organic compounds in the translocation ot trace metals through soils is governed by their chemical be-

i n \ : k \ . n 14 Ml

Khan a i.. Hltcct ot some orcanic compounds on the niobiiii> of some trace metals through soil amended tu

0.20 i A .

0.J5 i .

O.JO I

o.os

0.00 L 0 .20

FOICMIC ACID • CITKACO) HTlHASOi.

• \

•0 i^o> )

•o A,

I

"6^ o

• — » •t—A - » — «

L cnuMOL

0 . 1 5 o-

S 0 . 1 0 ^"S:.A>

J- 0 .05

0.00

ACTTONI

\

' A . • a .

0.15 A .

rORMALOtBYBE ACtTALMHYM I

••v^ A-.^ °->^

MSnULEOWATU

\ .

0 00 L 0 . 0 5 . 0 ) 0 . 0 0 .0 5 . 0 10 .C 0 0

Coocmvation of Fly ash (g Ag) s.o

FIR. 1. Mohilin of trace metals as affected by .some organic compounds ihrouch .soil amended with flv ash 1-1 Fh

1 0 . 0

(O) Co. ( • ) Zn. (Al Cu. AE,

haviour in soils. These organic compounds indicate the mobility order in the various soil systems as formic acid > acetic acid > methanol > formaldehyde > ethyl methyl > ketone aceialdehyde ethanol. The highest mobility in case ot formic acid and acetic acid seems to be governed by their ionizing as well as solubilizing tendencies as revealed by their inverse order of pK.^ values. Hov^ever. in case of alcohols, the mobility metal ions is governed by the molecular size and the capability of forming the hydrogen bond with the hydrated metal ions in soil of pH 8.4. Thus the metal ions move in the order :

/ H OH-CH? H 0-H-C:Hs M-'-(l > M - - - 0

-OH-CH. -H-C:H5

H>dijieJ metal ions Hydraled nieial ions meihanol complex ethand complex

which is the rexerse sequence of the molecular weight of the given alcohols.

The mobility of metal ions in case of organic com

pounds containing carbonyl groups follows the order : acetone > formaldehyde > ethyl methyl ketone > acetal-dehyde. The order of mobility seems to be governed b\ polarity of these compounds in soils. The carbonyl compounds containing alkyl groups capable of imposing steric hindrance also play a decisive role in determining the at-finit> of interaction of metal ions v\ ith these organic compounds used as mobile phase. For example, in ca>e of ketones, the mobility of metal ions is higher in acetone than in ethyl methyl ketone since the carbonyl group in acetone is sterically less hindered as compared to ethyl methyl ketone. Similarly, in aldehydes, the mobility of metal ions in formaldehyde is more as compared to acetal-dehyde due to the steric hindrance posed by the one meihv 1 group attached over the carbonyl group of acetaidehyde.

Experimental

The soil used vxas an "aerie Halaquept" (fine sandy loam) collected from Aligarh Muslim University, Aligarh (U.P.). The physicochemical properties of the soil and fl> ash were : soil : sand 60%. silt 26%, clay 14%, organic

j i I \ :K 11 I 4 MI

J Indian Chem Soc, Vol. 77. JuK 2{KKJ

HKiticr 1)45',. pH 84 (1 ; 2.5. soil : water). EC 0.167 niniho >.'m . CLC 16.. cmol (p*) kg"' soil, exchangeable Nil- in , K- 0 5 Ca-^ 3.5 and Mg-" 1.2 cmol (p* i kg ' ; fl\ ash pH (1.65 (1 : 2.5 FA : water ratio). E.C. 0.71 mmho cm-', axailable NH^-N 30 4, NO^-N 24.80. NO;-N 3.20. P 4''() and K 198.4 mg kg"' fly ash (determined by the meihoJ lived in soil anal).sis)".

The M'll sample was dried, ground and then pa.ssed tht>ugh a l(K) mesh sieve'- to make it homogenous. Fh ash was added to the soil at the rate of 0, 2.5. 5.0. 7.5. 10.0. 12 5 g'kj: Mill. It was then thoroughly mixed so as to get a homogenous mixture in each case. The slurry of the amended soil was then prepared in distilled water and coaled on glass plates with an uniform layer of 0.5 mm thickness m each case. The plates were then air-dried at room temperature (30± 3"). Two lines were drawn at 4 and 14 cm from the base on the plate so as to allow developer to run a distance of 10 cm for the study. An aqueous solution of Co. Zn. Cu, Ag and Pb nitrate (0.01 M) were spotted on the base line. The plates were wrapped with the wet filter paper at the bottom and developed by ascending chromatography in closed T L C chamber using distilled water and aqueous solution (0.25 M) of organic compounds, viz formic acid, acetic acid, methanol, ethanol. tormaldehvde. acetaldehvde. acetone and ethyl methyl ketone a a developer. The developed plates were air-dried at 6( and spots were detected by spraying 0.1 (w/vi elhanolic solution of haemotoxylin that gave stable violet spots tor Co--. Cu--. Zn-*. Pb-* and red for Ag* and the y?, values evaluated.

Ri'ferenci's

I F E Allison. Soil Organic Metal and iis Role in Crop Produc

tion'. EKcvii-r. Ni-« ^ork. W^

D T Gibson and M IXvkci. MuroNa! tX-frsdatiun of Orpani. Compounds'. Acadiiinc. New Vori l">4. I Arisukeich and Z Solaiiian. Chen, Ahsii . 19* ". 127. - : i

J P Sinf hal. .S l Khan and .S Nandan Indian J Af;ni Si i 1979. 49. 42.1

S U Khan and N N Khan./'/<J':» .v •. . I9S.';. 74. . 87. F A Ba.-zaz, R W CailsonanJG L Kolfi. A' .r,.-; Pollul.. \914.1. 2~\ D C Gupta. G Smfh. R N . Savcn. and K C Amarjccl. £/;•./.•.•» Afcdiu. 1998. .V:57, W H .Smilh. £/..ir,.r: Pollul. \9'7-i. (, l i i . S V Khan. D Nandan and N N Khar. L- unm. Pollut. I9i>; A

119

P. C Brook and S P McGralh. J S •:. Sn. 1984. 35. .141. P C Brook. S P. McCrath. D A Klein and E i Ellioi. "Environmcn!.-il Conlaminalion", CEP Ltd.. Edinburc. IvS-;

S. U Khan. T. Begum and J Singh. In^nu-; J Environ. Hlili . 19VfS. 38. 41; C O Plan. Chen: .A/'sir . IV"'-. 81. 250; Andojumpei. N Jungi. Uzavva. Nagano and Yoji. Sip:'-'i. Di-jn Hirynfiaku 2I/M//( 1986. 57. .191 (C/jf"J Ah\ir. 198". 107. b:Oi. A. Majumdar and S N. Mukherjee, Fuel Sa Te. hm>!.. IVS.'S. 1. 80. L. C. Mishra and K. N Shukla. Environ PoUui. Sei A. 19i.6. 41. 1.

Gaszczyk. Pvszark and Paiizko. En\in.r s,i Res...l99b. 51. ?2''

F. C Slevenson and M S Ardakani. Micronuuients in Agncui lure". Soil Sci Soc of Amenca. Madi ^ n. 19"2: L. J. LunJ A t. Page and C. O. Nel.son. ./ Enviroi: Q.,.r. 1976, 5. .1.10

A. K Sen and A. K. Dc. Wat Rey. 19>-. 21. 885; Indian J Id h

nol.. 198". 25. 259

A V\'alklev and C A Black. Sml .<;,, 1 -4-. 63. 251.

M L. Jackson. "Soil Chemical Anaivsiv". Prenlicc-Hal! Englevkood Cliffs. 19.';8

S. U Khan. M A Qurohi and J Sincr. J Environ Hlih . Ivv". .19. 217,

S V. Khan. N. N. Khan and N lijbal. Er.-.mm. Pollul.. 1991. 70 109

328

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Dear Dr7Mr/Ms^. Editor thanks for your manuscript entitled.

( R e r N o Z P ^ 7 - ^ - ^ J " - ^ ' ! S r ^ -

submitted for publication in .\ .!.'..'...f7*rf?.--. .)

Authors should invariably subscribe the journal, please subscribe ihe journal immediately by sending the subscription amount mentioned below. Please ignore if \ou have subscribed or your are a life member. Editor is pleased io inform vou that your caper ii ;>ccepied for publication m xX^.'.l..'...'^~r?..': intheyear.....^^.^':^. ". issue No I A). ^. Authors have to buy a minimum of fifty reprints and p-V P^g^ charges if number of pages exceeds ..\.~. Accordingly kindly remit Rs./ySSr. ).6.oO./rrr. towards reprints and RsYUS S..

towards extra page charges latest by ^..\.'.^."..?.^.?.9 .so that your paper can be sent to the press. All remittances should be drawn in favour of Enviromedia by M.07 D.D. Add Rs. 30/- w ith outstation cheques. Thanking You & with regards

irs Sincere IV

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SUBSCRIPTIOS' CHARGES FOR 1999 & 2000

1. POLLUTION RESEARCH ^S^-eo INDIVIDUAL Rs. 300.00 ORGANIZATONS Rs. 640.00

2. JR. OF INDUSTRLVL POLLUTION CONTROL INDIVIDUAL Rs. 125-00 ORGANIZATIONS Rs. 275-00

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NAiiONAL BUREAIU OF SOIL SURVEY AND LAND USE PLANNING ^ ; ^ ~ ^ " * * " Amravati Road, NAGPUR - 440 010, India "" J4

tns-TMiiasuiN • i i i i » i i i m i II

•CAR

M3 Dr. D.K. PAL 08 May 2000 PHudpal SdaOist A Head Division of Soil Resource Studies

To, Dr Samiullah Khan, Dept. of Applied Chemistry, Faculty of Engineering & Technology, Aligarh Mulsim University, Aligarh 202 002.

Dear Dr. Khan,

I hope you have received my earlier letter intimating you about the status of your paper. I have just received the comments of the Reviewer. On the basis of his comments your paper " Mobility of Trace metals as affected by some Surfactants" by Khan etal has been accepted for pubUcation in the 1* issue of Clay research otf the year 2000. Please indicate if you want to order for r^erte. fz^-bvicJr

With best regards,

Yours wffckrely.

(O.K. PAL) Editor, Clay Research

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