Chemosphere 49 (2002) 545557

www.elsevier.com/locate/chemosphere

Heavy metals in agricultural soils from Piedmont, Italy.

Distribution, speciation and chemometric data treatment
O. Abollino a, M. Aceto b, M. Malandrino a, E. Mentasti
F. Petrella c
b

a,*

, C. Sarzanini a,

a
Department of Analytical Chemistry, University of Torino, Via Giuria 5, 10125 Torino, Italy
Department of Sciences and Advanced Technologies, University of East Piedmont, Spalto Marengo 33, 15100 Alessandria, Italy
c
Institute for Wood, Plant and Environment (I.P.L.A.), Corso Casale 476, 10132 Torino, Italy

Received 8 February 2002; received in revised form 18 June 2002; accepted 28 June 2002

Abstract
The distribution and speciation of heavy metals in ve agricultural soils of Piedmont Region (north-western Italy)
were investigated. Ten metals, namely Al, Cd, Cr, Cu, Fe, Mn, Ni, Pb, Ti and Zn were considered. Analytical determinations were performed by atomic emission or atomic absorption spectroscopy after microwave sample dissolution in
acid solution. Total metal concentrations t in the typical concentration ranges for unpolluted soils, with the exception
of cadmium and lead content in some horizons. The eect of sampling depth on concentrations was discussed. Speciation studies were carried out by applying Tessiers procedure, which allows to subdivide the total metal content into
ve fractions, representing portions bound to dierent components of the soil. Moreover, the element labilities in two
soils were evaluated by extraction with EDTA. Correlations among the variables and/or similarities among the sampling points were identied by principal component analysis and hierarchical cluster analysis.
2002 Elsevier Science Ltd. All rights reserved.
Keywords: Metals; Speciation; Agricultural soils; Pattern recognition; Piedmont; Italy

1. Introduction
The concentrations of heavy metals in soils are associated with biological and geochemical cycles and are
inuenced by anthropogenic activities, such as agricultural practices, transport, industrial activities, waste
disposal (Lund, 1990). The knowledge of both the total
concentration and chemical speciation is necessary to
characterise the behaviour of heavy metals in soils. In
fact it is well known that metals are present in soils in
dierent chemical forms, which inuence their reactivity
and hence their mobility and bioavailability. Speciation

Corresponding author. Tel.: +39-011-6707625; fax: +39011-6707615.

E-mail address: edoardo.mentasti@unito.it (E. Mentasti).

studies are usually carried out by single or sequential

extractions with reagents having dierent chemical properties (Rauret, 1998). These procedures partition metals
into fractions bound to dierent soil components. The
eect of the extractants simulates the inuence of differing environmental conditions, such as acid rain, on
the hypothetical release of metals from soil. Tessier et al.
developed an extraction scheme which allows the division of the total metal content into ve fractions: exchangeable, carbonate bound, iron/manganese oxide
bound, organic bound and residual fraction (Tessier
et al., 1979, 1980). The scheme was developed for sediments, but it can be applied to soils as well. Similar
procedures mainly dier in the extraction conditions
used. For example, F
orstners method (Calmano et al.,
1982) is based on conditions that allow the iron/manganese oxide fraction to be subdivided into three fractions

0045-6535/02/$ - see front matter 2002 Elsevier Science Ltd. All rights reserved.
PII: S 0 0 4 5 - 6 5 3 5 ( 0 2 ) 0 0 3 5 2 - 1

546

O. Abollino et al. / Chemosphere 49 (2002) 545557

(easily reducible, moderately reducible and nonsilicate

iron). Another commonly used speciation scheme is the
one developed by Community Bureau of Reference
(BCR), in which three fractions are identied: (I) exchangeable, water and acid soluble; (II) reducible; (III)
oxidisable species (Davidson et al., 1999).
The results obtained with extraction procedures are
of course operationally dened, because redistribution
and adsorption phenomena usually take place during
the treatments. Anyway the information obtained from
this kind of speciation study is of great value in order to
assess metal reactivity and hence their availability to the
environment and their potentially harmful eects.
In this study the total content of heavy metals and
their speciation were investigated in ve agricultural
soils from Piedmont, Italy. The following elements were
determined: Al, Cd, Cr, Cu, Fe, Mn, Ni, Pb, Ti, Zn.
Tessiers extraction scheme was used in the investigation
of the mobility and transport of the metals. Leaching
tests with Ethylenediaminetetraacetic acid (EDTA) were
applied to two soils. The results obtained were elaborated with pattern recognition techniques.

others come from uvial terraces. The samples were

collected at dierent depths, corresponding to the horizons shown in Table 2. A soil layer in the central part of
each horizon was sampled, e.g. sample PA for horizon
Ap (020 cm) corresponds to the 515 cm layer. The
depth proles of Poirino, S. Maurizio Canavese and
Carmagnola consisted of three samples, whereas those
of Vigone 1 and Vigone 2 consisted respectively of seven
and ve samples. The layer between 120 and 160 cm in
Vigone 1 is made of two bands, coloured red and grey
respectively; probably the rst one (sample V1E) has a
greater content of Fe(III) compounds and the other
(sample V1F) of Fe(II) and/or Cr(III) compounds. It is
therefore interesting to analyse them separately. The
code used to identify each sample refers to its origin (C
for Carmagnola, M for San Maurizio Canavese, P for
Poirino, V1 and V2 for Vigone 1 and Vigone 2) and to
the sampling depth (A for the rst horizon, B for the
second and so on).
Samples were collected with plastic tools and stored
in polyethylene bags; in-depth samples were obtained
from holes dug with a motor-driven excavator.
2.2. Apparatus and reagents

2. Experimental
2.1. Sampling
Soil samples were collected in ve sites in Piedmont
Region (north-western Italy). The sites are located in the
small towns of Poirino (Province of Asti), Vigone (two
sites, hereafter called Vigone 1 and Vigone 2), S. Maurizio Canavese and Carmagnola (Province of Torino).
Table 1 shows their taxonomic classication (Soil Conservation Service USDA, 1984; Soil Survey Sta, 1990;
Soil Survey Sta, 1999).
All soils were used as permanent meadows, with the
exception of the one from Carmagnola, cultivated to
maize. Vigone soils have an alluvial origin, whereas the

Metal determinations were performed by atomic

emission or atomic absorption spectroscopy depending
on the concentration level. In both cases calibrations
were performed with standard solutions prepared in
aliquots of sample blanks.
Higher concentrations were determined with a Varian Liberty 100 Inductively Coupled Plasma-Atomic
Emission Spectrometer (ICP-AES). Metal concentrations close to or lower than the detection limits for ICPAES were determined with a Perkin Elmer 5100
Graphite Furnace Absorption Spectrometer (GF-AAS)
equipped with a Zeeman-eect background correction.
H2 O2 present in the fourth fraction of Tessiers speciation scheme can damage the graphite furnace, there-

Table 1
Soil identication and description
Code

Position

Soil phase

USDA taxonomy

Carmagnola

Mezzi Po, coarse-loamy, typical phase

Typic Udiuvent, coarse-loamy, mixed,

mesic, calcareous

S. Maurizio Canavese

Foglizzo, coarse-loamy over sandy-skeletal,

little deep phase

Dystric Eutrudept, mixed, nonacid, mesic

Poirino

Poirino, ne-loamy, phase redoximorphic

Oxyaquic, maplustept ne-loamy, mixed,

nonacid, mesic

V1

Vigone 1

Quintanello, coarse-loamy, typical phase

Dystric Eutrudept, coarse-loamy, mixed,

nonacid, mesic

V2

Vigone 2

Fontanette, coarse-loamy, typical phase

Aquic Dystrudept, coarse-loamy, mixed,

nonacid, mesic

O. Abollino et al. / Chemosphere 49 (2002) 545557

547

Table 2
The general characteristics of the investigated soils
Soil parameter
Horizon

Particle size distribution

% sand
(20.05 mm)

% silt
(0.0050.002 mm)

pH

%C

Organic matter

CEC

% clay
(<0.002 mm)

CAa
CBa
CCa

Ap
AC
C

14.8
3.5
6.1

43.1
66.1
48.9

8.5
7.0
6.7

7.9
8.1
8.0

1.16
0.55
0.39

2.00
0.95
0.67

10.1
8.50
5.20

MAa
MBa
MCa

Ao
Bw
Cr

36.7
52.2
77

56.5
37.9
14.8

6.8
9.9
8.2

5.9
6.6
6.5

2.11
0.69
0.06

3.64
1.19
0.10

14.0
9.20
9.20

PAa
PBa
PCa

Ap
Bg
BCg

45.6
49.6
45.1

51.4
30.4
29.1

3.0
12.9
15.4

6.7
6.5
6.8

1.92
0.07
0.08

3.31
0.12
0.14

17.2
11.5
8.20

V1Aa
V1Ba
V1Ca
V1Da
V1Ea

Ap
AB
Bw
BC
C1

47.8
45.6
46.7
54.4
68.3

43.8
42.4
31.9
28.8
23.3

8.4
12
21.4
16.8
8.4

6.0
6.6
6.7
6.9
6.7

0.82
0.23

1.41
0.40

9.45
8.51

V2Aa
V2Ba
V2Ca
V2Da
V2Ea

Ap
AB
Bg
BC
C

60.5
67.9
83.4
97.2
95.3

34.4
23.2
14.6
2.2
3.2

5.1
8.9
2.0
0.6
1.5

6.2
6.5
7.4
7.4
7.3

0.87
0.27

1.50
0.47

9.32
12.1

Carmagnola: CA (040 cm); CB (4090 cm); CC (90120 cm); S. Maurizio Canavese: MA (030 cm); MB (3060 cm); MC (6080
cm) Poirino: IA (020 cm); IB (2080 cm); IC (80160 cm) Vigone 1: V1A (030 cm); V1B (3050 cm); V1C (5070 cm); V1D (70120
cm); V1E (120160 cm) Vigone 2: V2A (035 cm); V2B (3575 cm); V2C (75120 cm); V2D (120175 cm); V2E (175195 cm).

fore the extract was pretreated (1:1) with a solution of

ascorbic acid to eliminate H2 O2 in excess. The addition
of the reducing agent did not cause a contamination of
the extract.
Cadmium and lead in extracts from Tessiers second
fraction were determined by adsorptive cathodic stripping voltammetry (ACSV) with an Autolab PGSTAT10
voltammetric analyser connected to a Metrohm 663VA
stand equipped with a hanging mercury drop electrode.
A Milestone MLS-1200 Mega microwave laboratory
unit was used for the dissolution of the samples. High
purity water (HPW) produced with a Millipore Milli-Q
system was used throughout. All the reagents used were
of analytical grade. Standard metal solutions were prepared from concentrated stock solutions (Merck Titrisol).
2.3. Procedures
The granulometric analysis was executed according
to standard procedures (Gee and Bauder, 1986; Sheldrick and Wang, 1993). All analyses were performed in
triplicate. Blanks were simultaneously run. Soil samples
were air-dried, passed through a 2-mm stainless steel

sieve and ground in a centrifugal ball mill in order to

homogenise them.
The analytical accuracy of the procedure utilised to
determine the total metal concentrations in investigated
soils, was conrmed by analysis of a standard reference
material (Estuarine Sediment CRM 277, BCR UE
Brussels); instead, the accuracy of the sequential extraction procedure was obtained by comparing, for a
few samples, the total metal content with the sum of the
metal percentages extracted in the ve fractions after
digestion (HF/aqua regia) and analysis of the fth
fraction.
2.3.1. Sample dissolution
For the determination of the total metal concentrations acid digestion in a microwave oven was chosen as
the dissolution procedure.
Sample aliquots of 100 mg were treated with a mixture of 5 ml of aqua regia and 2 ml of hydrouoric acid
in tetrauoromethoxyl (TFM) bombs. Four heating
steps of 5 min each (250, 400, 600, 250 W power respectively), followed by a ventilation step of 25 min,
were applied. Then 0.7 g of boric acid were added, and
the bombs were further heated for 5 min at 250 W and

548

O. Abollino et al. / Chemosphere 49 (2002) 545557

again cooled by a ventilation step of 15 min. At the end

of the full treatment, the samples appeared completely
dissolved. Finally the resulting solutions were diluted to
100 ml with HPW. The solutions were directly employed
for the ICP-AES and GF-AAS analysis.
2.3.2. Measurement of pH, organic carbon, organic
matter, cation exchange capacity
pH, organic carbon, organic matter and cation exchange capacity (CEC) were determined according to
the ocial methods of soil analysis of the Italian legislation issued in 1992, as described below. While research
was in progress, new ocial methods were issued in
which a dierent procedure for pH measurement was
xed, whereas the methods for organic carbon, organic
matter and CEC determination were not changed.
A 1:2.5 soilwater suspension (8 g of soil for 20 ml of
HPW) was prepared and left standing overnight for pH
measurement.
The organic carbon was determined by Walkley
Black method (Soltner, 1988). It was oxidised to carbon
dioxide with potassium dichromate in the presence of
sulphuric acid. The unreacted potassium dichromate
was titrated with iron(II) sulphate.
Considering that the average content of carbon in
soil organic matter is equal to 58%, the conversion factor 1.724 was used to calculate the percentage of organic
matter from the content of organic carbon.
The CEC was determined with barium chloride
(Bartels, 1996). The samples of soil were saturated with
Ba by treatment with barium chloride solution buered
to pH 8.2. The addition of magnesium sulphate caused
the formation of insoluble barium sulphate and, therefore, the exchange Ba/Mg. The barium in excess left in
solution was determined by complexometric titration
with EDTA.
2.3.3. Sequential extraction procedure
Extractions were carried out on 1.0 g aliquots of soil
and involved the ve following steps (Tessier et al., 1979,
Tessier et al., 1980):
1. Exchangeable fraction: the sample was placed in contact with a high ionic strength solution, in order to
release the so-called exchangeable fraction of metal
traces by altering the sorptiondesorption supercial
processes. In this step 8 ml of 1 N MgCl2 were added
to the sample and the suspension was shaken for 1 h.
2. Fraction bound to carbonates: the fraction of metal
traces bound to carbonates, present in the sample,
may be selectively labilised by varying the pH of
the sample itself with a slightly acidic extraction solution. 8 ml of 1 M of CH3 COONa, plus CH3 COOH
(pH 5) were added to the residue obtained from the
rst extraction, and the suspension was shaken for
5 h.

3. Fraction bound to iron and manganese oxides: this

fraction may be labilised in anoxic reducing conditions. Therefore, 20 ml of 0.04 M NH2 OH
HCl in
25% CH3 COOH were added to the residue and the
suspension was shaken for 6 h at the temperature
of 96  3 C.
4. Fraction bound to organic matter and to sulphides:
this fraction can be released by treating the sample
with an oxidising agent. Three ml of 0.02 N HNO3
and 5 ml of 30% H2 O2 were added to the residue obtained from the third extraction, and the suspension
was shaken for 5 h at the temperature of 85  2 C.
After cooling, 5 ml of 3.2 M CH3 COONH4 were
added to the suspension, which was diluted to 20
ml with HPW and shaken for 30 min.
5. Residual fraction: it is the metal fraction present as
scatter within the crystal lattice of the rocks and minerals that constitute the soil. It was calculated from
the dierence between the concentration of total
metal and the sum of the rst four fractions.
After each extraction the suspension was subjected
to centrifugation for 20 min at 4000 rpm. The solution
was separated, while the precipitate was washed with
10 ml of HPW and centrifuged again for 5 min. The
washing water then was added to the surnatant, while
the precipitate was used for the subsequent extractions.
The extracts were diluted to 50 (rst two fractions) or
100 (next two ones) ml, stabilised by addition of 50 or
100 ll of concentrated nitric acid respectively and
analysed.
2.3.4. EDTA-extractable fraction
This fraction was determined according to Lakanen
Ervi
o procedure (Lakanen and Ervi
o, 1971). Twentyve ml of extracting solution (0.02 M EDTA in 0.5 M
CH3 COONH4 ) were added to aliquots of 5 g of soil; the
suspensions, as obtained, were shaken for 30 min, ltered and analysed.
2.3.5. Voltammetric analysis
Aliquots of 10 ml of the samples were added with a
2  103 M solution of 8-hydroxyquinoline. The deposition potential was )0.30 V for Pb and )1.10 V (start
potential )0.10 V) for Cd and the deposition time was
120 s.
2.3.6. Chemometrics
PCA and HCA were performed with XLStat 4.4
software package, used as a Microsoft Excel plug-in.
When concentrations were below the detection limit, a
random value between zero and that limit was inserted
in order to be able to thoroughly apply PCA and HCA
without losing any case.

O. Abollino et al. / Chemosphere 49 (2002) 545557

3. Results and discussion

3.1. Granulometry and general characteristics
Table 2 reports particle size distribution and some
general characteristics, namely pH, percentages of organic carbon, organic matter and CEC, of the soils examined in this study. In general, all soils have a low
content of clay and organic matter, so they tend to be
permeable. As it can be seen, the percentage of sand is
very high for Vigone 2B, whereas the soil from Carmagnola has a large silt fraction. The soils in S. Maurizio Canavese, Vigone 1 and Vigone 2 become more
sandy with increasing depth. The highest percentages of
clay were found in samples PB and PC and in two layers
(V1C and V1D) of Vigone 1.
pH values generally increase with depth, more pronouncedly for Vigone 1 and Vigone 2, and slightly for
the other soils. The content of organic carbon and organic matter always decreases with increasing depth.
Also CEC follows this trend, with only one exception.

549

If the top soil is considered, pH increases in the

order S. Maurizio Canavese < Vigone2 < Vigone1 <
Poirino < Carmagnola; San Maurizio Canavese and
Poirino are the richest in organic matter and have the
highest CEC, whereas samples V1A and V2A have the
lowest values of these parameters.
3.2. Total metal concentrations
Table 3 reports the total metal concentrations in the
investigated soils and, for comparison, their typical
concentration ranges in soils (Alloway, 1990). The examined soils can be considered unpolluted, since their
concentrations t in the typical ranges, with the exception of cadmium in Poirino, S. Maurizio Canavese and
Carmagnola and lead in some horizons of S. Maurizio
Canavese, Vigone 1 and Vigone 2.
The soil from S. Maurizio Canavese has the highest
content of chromium and nickel; probably because of
the nature of the parent material, which consists of ultramac rocks (Alloway, 1990; McGrath and Smith,

Table 3
Total metal concentrations in considered soils, typical literature ranges and most common values, average abundance in the earths
crust (values in mg/kg unless otherwise stated)
Al

Cd

Cr

Cu

Fe

Mn

Ni

Pb

Ti

Zn

CA
CB
CC

67 900
77 800
74 700

5.4
6.4
3.4

162
174
146

30
44
28

34 000
39 500
36 300

833
1080
899

102
141
132

215
291
229

3550
4160
3760

94
103
86

MA
MB
MC

73 500
72 600
57 900

4.9
4.3
2.1

588
562
598

34
28
30

46 100
44 600
41 800

1130
965
802

675
535
510

270
99
750

5520
5250
6150

104
73
73

PA
PB
PC

47 400
81 500
77 800

2.7
4.3
5.1

124
170
143

19
46
36

22 900
38 600
36 200

525
1050
900

32
126
94

141
181
149

3070
4270
3850

82
92
93

V1A
V1B
V1C
V1D
V1E
V1F

58 600
69 300
72 100
66 200
56 600
67 600

1.3
1.1
0.9
0.4
1.3
0.3

116
162
156
137
138
1580

24
27
36
34
30
34

31 600
24 400
42 500
40 600
63 700
33 800

585
670
1410
1030
1260
360

74
96
104
113
130
111

374
350
582
354
447
392

4210
4580
4280
3980
3800
4060

73
74
80
75
69
79

V2A
V2B
V2C
V2D
V2E

82 100
75 300
76 200
75 500
63 800

0.8
1.6
0.7
0.9
0.7

140
150
115
118
105

45
33
24
17
16

51 200
40 900
34 500
37 800
32 200

877
897
639
623
485

134
123
95
82
81

524
302
213
204
168

4100
3930
3690
3630
2710

135
81
71
61
57

0.012.0

51500

2250

7000
42 000

2750

2300

1900

0.11

70100

2030

20
10 000
1000

50

1030a

50

Typical
range
Common
value

a
These values are for rural soils. The common values for lead in urban soils are reported as 30100. Carmagnola: CA (040 cm); CB
(4090 cm); CC (90120 cm); S. Maurizio Canavese: MA (1020 cm); MB (4050 cm); MC (6080 cm); Poirino: IA (depth 515 cm); IB
(depth 4050 cm); IC (depth 100120 cm); Vigone 1: V1A (030 cm); V1B (3050 cm); V1C (5070 cm); V1D (70120 cm); V1E (120
160 cm); V1F (120160 cm); Vigone 2: V2A (035 cm); V2B (3575 cm); V2C (75120 cm); V2D (120175 cm); V2E (175195 cm).

550

O. Abollino et al. / Chemosphere 49 (2002) 545557

1990). Since Vigone soils are in open country, far from

main roads, trac cannot directly explain the high lead
concentrations found, which could be derived from
the parent material or, especially in the case of Vigone
2, to atmospheric deposition deriving from a distant
source.
As to the vertical proles, a decreasing trend in the
concentrations of all elements with sampling depth
can be observed in Vigone 2. All metals are depleted in
the top layers of Poirino and Carmagnola soils; this
behaviour can be explained for Carmagnola with the
uptake of metals from maize plants. A rather high concentration of chromium was found in the grey coloured
band in Vigone 1; this result conrms the hypothesis
made when samples were collected. No other remarkable features or trends can be observed in the metal
contents of the ve soils.
3.3. Speciation
3.3.1. Tessiers extraction
Metal speciation according to Tessiers scheme was
performed on all samples. The analyte percentages in the
ve fractions are shown in Tables 48. It must be
pointed out that the low percentages reported for some
elements correspond to meaningful measured concentrations: for instance a percentage of 0.003% of Al extracted from sample PC in the rst fraction corresponds

Table 4
The percentages of the extracted metals in the rst fraction of
Tessiers speciation scheme
Al

Fe

Mn

Ti

CA
CB
CC

<0.001
<0.001
<0.001

<0.001
<0.001
<0.001

0.06
0.05
0.08

0.004
<0.004
0.005

MA
MB
MC

0.008
0.002
0.001

0.001
<0.001
<0.001

8.7
4.3
3.0

0.01
0.01
0.01

PA
PB
PC

0.004
<0.001
0.003

0.005
<0.001
<0.001

10.9
4.9
2.6

0.01
0.02
0.02

V1A
V1B
V1C
V1D
V1E
V1F

0.003
0.002
0.002
<0.001
0.001
0.001

0.004
0.001
0.002
<0.001
<0.001
0.002

1.6
0.2
0.06
0.09
0.02
0.06

<0.004
<0.003
<0.003
<0.004
<0.004
<0.004

V2A
V2B
V2C
V2D
V2E

<0.001
0.001
<0.001
<0.001
<0.001

0.001
0.003
0.003
0.002
0.003

0.05
0.03
0.02
0.01
0.03

<0.004
<0.004
<0.004
<0.004
<0.005

to 2.24 mg/kg. As remarked in the Introduction, this

scheme, like almost all existing speciation procedures,
gives operationally dened results.
The rst fraction contains negligible concentrations
of almost all of the metals considered, except Al, Mn
and sometimes Fe and Ti. In Carmagnola and in most of
the Vigone 2 samples also the exchangeable concentrations of Al are undetectable. The extracted percentages
of Al, and especially Mn, in this fraction, are higher in
Poirino and S. Maurizio Canavese than in the other
soils. The extracted fractions of Al and Mn decrease
with increasing depth in most cases.
Most of the considered metals are present in the second fraction, even if at low percentages. The concentrations of the extracted metals in Poirino and
S. Maurizio Canavese decrease with increasing depth,
with a few exceptions. Chromium was detected only in
S. Maurizio Canavese and in a few other samples
whereas its concentration in the other soils was lower
than the detection limit. The percentages of major elements (Al, Fe and Ti) are lower in Vigone 1 than in the
other soils.
Relatively high extracted percentages of most metals
were found in the third fraction. The behaviour of the
elements diers from soil to soil: for instance, copper
was more extensively extracted from Carmagnola and
Vigone1, iron from Poirino and S. Maurizio Canavese
and nickel from S. Maurizio Canavese soil. No regular
trend as a function of depth can be observed.
The concentrations of the metals in the fourth fraction are not higher than in the third one, and in many
cases are lower. Therefore a relatively small fraction
of the metals is bound to organic matter and the highest percentages are associated to the mineral matrix of
the soil. As expected, the samples with higher organic
matter contents have higher levels of most metals in
this fraction. The concentrations of major elements (Al,
Fe and Mn) in Vigone 1 and Vigone 2 are frequently
lower than in the other investigated soils. The metal
content in the fourth fraction generally decreases
with increasing depth, probably because organic substances are less present in the deep levels, because the
biological activities mainly occur in the supercial horizons.
The majority of the metals present in the soils is
contained in the fth fraction, i.e. they are strongly
bound to the soil matrix. An exception to this trend is
represented by Mn, which is also present at high percentages in the third fraction. The most inert element
was found to be Ti.
The presence of ferric and ferrous compounds respectively in E and F levels of Vigone 1 soil does not
seem to inuence the extractability of iron, or of other
elements, since there are not remarkable dierences between the percentages found in the two levels, whose
behaviour is similar to the one of the other horizons.

O. Abollino et al. / Chemosphere 49 (2002) 545557

551

Table 5
The percentages of the extracted metals in the second fraction of Tessiers speciation scheme
Al

Cd

Cr

Cu

Fe

CA
CB
CC

0.03
0.04
0.02

3.30
3.61
10.12

<0.15
<0.14
<0.17

2.63
3.20
2.56

0.03
0.07
0.09

MA
MB
MC

0.05
0.03
0.04

2.10
<0.06
<0.12

0.22
0.08
0.06

3.71
1.73
1.73

PA
PB
PC

0.05
0.03
0.02

5.37
0.39
0.23

<0.20
<0.15
<0.17

V1A
V1B
V1C
V1D
V1E
V1F

0.01
0.01
0.01
0.01
0.01
0.01

0.02
0.06
0.07
0.13
0.02
0.08

V2A
V2B
V2C
V2D
V2E

0.03
0.06
0.02
0.05
0.06

0.02
<0.01
0.04
0.04
0.01

Mn

Ni

Pb

Ti

Zn

6.11
7.99
9.38

1.88
1.61
0.81

1.15
1.10
0.97

0.02
0.01
0.01

<0.01
<0.01
<0.01

0.04
0.02
0.02

4.21
2.92
2.16

0.61
0.58
0.44

4.19
0.49
0.07

0.01
<0.01
<0.01

2.11
0.39
0.21

2.67
0.86
1.26

0.11
0.02
0.01

19.24
3.98
2.13

3.25
0.82
0.77

2.23
0.62
2.52

0.01
0.01
0.01

4.34
0.21
0.20

<0.21
<0.15
<0.16
<0.18
<0.18
<0.02

7.58
2.78
2.16
1.26
1.32
1.56

0.01
0.01
0.01
0.05
0.01
0.01

0.98
0.51
0.20
0.26
0.21
1.34

1.89
1.21
1.18
2.83
1.62
0.44

0.742
0.482
0.112
0.452
0.152
0.452

<0.01
<0.01
<0.01
<0.01
0.01
<0.01

0.09
<0.01
<0.01
<0.01
<0.01
<0.01

<0.18
<0.17
0.63
<0.21
<0.24

0.70
1.41
1.60
2.21
1.41

0.09
0.14
0.05
0.13
0.16

5.71
3.76
2.30
0.85
4.35

0.99
3.04
2.51
2.76
2.79

0.342
0.392
0.582
0.422
0.512

0.02
0.02
0.01
0.01
0.01

<0.01
0.71
<0.01
<0.01
<0.01

Table 6
The percentages of the extracted metals in the third fraction of Tessiers speciation scheme
Al

Cd

Cr

Cu

Fe

CA
CB
CC

<0.01
<0.01
<0.01

2.48
1.59
<0.15

0.97
1.01
1.28

15.77
23.64
30.00

7.26
8.82
9.71

69.15
79.17
79.87

1.81
1.37
1.08

3.86
2.57
3.19

0.02
0.03
0.02

15.85
6.25
7.51

MA
MB
MC

1.09
1.39
1.88

0.47
<0.12
<0.24

1.87
2.35
1.67

8.44
7.36
7.17

13.76
15.64
15.85

35.04
43.83
33.17

24.00
23.55
24.90

4.26
3.96
0.41

0.03
0.05
0.04

30.10
7.47
7.18

PA
PB
PC

0.97
1.01
0.83

0.74
0.39
0.10

0.90
1.92
1.85

10.63
8.61
10.47

21.73
16.28
15.93

54.67
94.76
52.44

18.06
7.09
6.03

4.85
4.35
4.12

0.03
<0.01
0.01

31.10
7.71
6.85

V1A
V1B
V1C
V1D
V1E
V1F

1.50
1.37
1.37
1.38
1.68
1.25

1.05
0.76
0.84
3.60
9.47
1.28

3.54
6.05
7.69
9.42
4.69
0.49

14.62
14.78
17.42
17.44
19.13
26.56

10.81
16.57
9.13
10.19
6.61
8.88

64.61
87.46
44.47
36.21
97.70
28.33

9.53
10.52
11.30
17.61
28.54
11.89

2.04
1.45
0.82
0.41
0.54
0.66

0.02
0.03
0.01
0.01
<0.01
0.01

15.21
7.46
7.50
7.92
10.93
11.53

V2A
V2B
V2C
V2D
V2E

0.37
0.45
0.52
0.54
0.13

0.96
1.16
2.86
3.17
1.39

1.74
1.79
2.30
1.47
3.05

2.29
8.82
16.13
15.82
21.38

3.41
5.11
6.42
4.95
7.02

11.52
36.68
46.79
28.73
31.55

3.51
7.03
9.28
8.10
9.11

0.12
0.31
0.56
0.34
0.33

0.02
0.01
0.04
0.02
0.06

3.31
3.41
2.61
1.37
5.30

3.3.2. Extraction with EDTA

The total pool of potentially available species in soils
can be estimated from the EDTA-extractable fraction

Mn

Ni

Pb

Ti

Zn

(Lund, 1990). Bioavailable concentrations are probably more correlated to the eect of milder extractants
such as neutral salts (Hani, 1990; Rauret, 1998). The

552

O. Abollino et al. / Chemosphere 49 (2002) 545557

Table 7
The percentages of the extracted metals in the fourth fraction of Tessiers speciation scheme
Al

Cd

Cr

Cu

Fe

Mn

Ni

Pb

Ti

Zn

CA
CB
CC

0.97
0.85
0.70

<0.09
<0.08
<0.15

1.64
1.33
1.18

28.57
9.95
10.00

0.74
0.45
0.30

7.19
11.67
7.09

1.04
0.77
0.50

1.07
0.88
0.81

0.08
0.04
0.05

0.42
0.43
0.33

MA
MB
MC

1.04
0.51
0.43

<0.10
<0.12
<0.24

3.79
2.90
2.16

17.41
3.02
3.77

0.28
0.03
0.04

4.96
2.55
1.42

<0.07
<0.09
<0.10

0.43
<0.50
<0.07

0.03
0.03
0.08

3.10
<0.01
<0.01

PA
PB
PC

1.18
0.26
0.24

<0.18
<0.12
<0.10

7.81
2.33
1.28

24.47
1.12
<0.35

0.92
0.05
0.02

12.97
17.14
9.77

<1.56
<0.40
<0.53

0.58
0.63
0.57

0.02
<0.01
0.03

4.16
<0.01
<0.01

V1A
V1B
V1C
V1D
V1E
V1F

0.66
0.39
0.63
0.62
0.27
0.42

<0.38
<0.44
<0.53
<1.38
<0.38
<1.46

5.68
3.31
2.97
3.35
2.19
0.14

11.21
1.63
1.59
2.13
<0.42
0.77

0.01
<0.01
<0.01
<0.01
0.01
0.01

3.50
0.82
0.19
0.36
0.05
0.14

2.80
1.99
2.35
2.89
1.87
2.01

<0.13
<0.14
<0.09
<0.14
<0.11
<0.13

0.18
0.05
0.01
<0.01
0.04
0.05

<0.01
<0.01
<0.01
<0.01
<0.01
<0.01

V2A
V2B
V2C
V2D
V2E

0.10
0.07
0.07
0.05
0.10

9.27
7.13
29.22
80.04
35.51

2.39
0.78
0.86
<0.42
0.94

3.53
1.08
<0.52
<0.73
<0.78

0.06
0.03
0.14
0.11
0.27

1.08
0.61
0.42
0.60
0.14

1.66
2.18
1.67
0.80
1.30

<0.09
<0.17
0.34
<0.24
<0.30

0.03
0.06
0.21
0.21
0.24

1.28
0.96
0.39
<0.01
<0.01

Table 8
The percentages of the extracted metals in the fth fraction of Tessiers speciation scheme
Al

Cd

Cr

Cu

Fe

Mn

Ni

Pb

Ti

Zn

CA
CB
CC

99.00
99.11
99.27

98.05
98.26
96.60

99.09
99.21
99.16

52.99
63.09
57.25

91.97
90.65
89.91

17.49
1.13
3.59

95.26
96.25
97.61

97.97
98.48
98.34

99.88
99.92
99.92

83.72
93.32
92.16

MA
MB
MC

97.81
98.07
97.65

99.11
99.94
99.97

98.04
98.22
98.70

70.29
87.73
87.32

85.92
84.31
84.09

47.03
46.43
60.21

75.39
75.86
74.65

97.04
98.49
99.84

99.93
99.90
99.86

64.70
92.13
92.61

PA
PB
PC

97.79
98.71
98.90

97.93
99.71
99.86

97.05
98.54
98.92

62.05
89.38
88.15

77.24
83.65
84.03

2.27
1.28
33.03

78.50
92.04
93.16

97.44
98.13
97.59

99.94
99.97
99.93

60.40
92.08
92.95

V1A
V1B
V1C
V1D
V1E
V1F

97.82
98.23
97.99
97.99
98.04
98.33

99.62
99.67
99.67
98.44
96.77
99.39

96.87
96.85
96.42
95.71
97.67
99.78

66.47
80.73
78.74
79.05
79.25
71.01

89.17
83.42
90.86
89.75
93.37
91.10

29.27
10.96
55.08
63.07
2.02
70.13

85.78
86.28
85.17
76.66
67.97
85.66

99.31
99.50
99.70
99.84
99.79
99.75

99.80
99.92
99.97
99.97
99.94
99.94

84.70
92.53
92.49
92.07
89.06
88.46

V2A
V2B
V2C
V2D
V2E

99.48
99.41
99.39
99.35
99.71

96.58
97.24
89.29
72.25
87.70

98.58
99.10
98.74
99.32
98.67

93.44
88.63
81.63
81.01
76.68

96.43
94.71
93.39
94.80
92.54

81.64
58.92
50.46
69.81
63.94

93.84
87.75
86.54
88.34
86.80

99.94
99.85
99.70
99.86
99.85

99.92
99.91
99.73
99.75
99.69

95.40
94.92
97.00
98.61
94.68

LakanenErvi
o procedure is usually applied only to
soils that have a pH value lower than 6.5. Anyway, it
was used also for two samples from Vigone 1 with a

slightly higher pH (6.70 and 6.76). The results obtained

are reported in Table 9. As expected, the percentages of
extracted metals are higher than those obtained with the

O. Abollino et al. / Chemosphere 49 (2002) 545557

553

Table 9
The percentages of metals extracted with EDTA
Al

Cr

Cu

Fe

Mn

Ni

Pb

V1A
V1B
V1C
V1D
V1E
V1F

0.15
0.15
0.08
0.10
0.08
0.07

0.20
0.07
0.12
0.10
0.13
0.01

16.12
4.22
2.54
1.74
1.96
1.06

1.20
0.75
0.25
0.38
0.17
0.21

63.59
31.79
9.99
18.74
33.09
7.81

6.39
5.76
1.94
2.21
5.48
0.50

1.17
0.93
0.54
0.99
0.54
0.84

0.10
0.07
0.05
0.03
0.02
0.02

4.84
0.98
0.76
0.59
1.05
0.41

V2A
V2B
V2C
V2D
V2E

0.12
0.09
0.04
0.02
0.03

0.25
0.10
0.06
0.04
0.09

9.09
5.76
2.92
1.45
3.79

0.65
0.52
0.22
0.11
0.28

42.99
8.53
6.59
6.90
39.79

3.35
2.06
0.53
0.62
4.38

0.78
0.97
1.05
0.65
1.52

0.04
0.04
0.01
<0.01
0.01

3.47
1.94
0.97
0.47
0.68

leaching test with water because EDTA is a strong ligand that is able to react not only with the more mobile
forms of the metal, but also with those more strongly
bound to organic or inorganic soil constituents. In fact
the percentages of the extracted metals found with this
procedure are comparable to those obtained in the
third and fourth fraction of the Tessiers procedure. The
order of extractability was Mn > Cu > Ni > Zn > Pb >
Fe > Cr > Al > Ti and does not seem to be tightly related to the values of the formation constants with
EDTA, which decrease in the order Fe(III) > Cu >
Ni > Pb > Zn > Al > FeII > Mn (Sillen and Martell,
1979). Extractability is probably dependent on the
chemical form of the metal in soil and therefore to its
mobility. The extracted percentages range from 0.017
to 68.59 in Vigone 1 soil and from 0.007 to 39.79 in
Vigone 2.

Ti

Zn

PC1 and PC2, were considered to represent all examined

objects and variables). The combined plot of scores
(coordinates of the objects on the new variables) and
loadings (weights of original variables on the linear
combination PCs are built from) allow us to recognise
groups of samples with similar behaviour and the existing correlation among the original variables.
In Cluster Analysis, samples are considered as objects
in an n-dimensional hyperspace (with n number of
variables), described by n-components vectors. The agglomerative hierarchical clustering procedure was used
to evaluate similarities among samples. In this way, a
dendrogram was obtained, in which correlations among
samples can readily be seen. This technique is complementary with PCA; the whole information is shown in
the dendrogram.
Only some PC and dendrogram plots will be shown
in this paper, and the other ones are available on request
from the authors.

3.4. Principal components analysis and cluster analysis

A chemometric study on the analytical data was executed in order to obtain a visual representation of the
main characteristics and of metal distribution in the soils
and to nd out similarities and correlations among
variables which would be more dicult to detect just
observing the numbers in the tables.
Principal components analysis (PCA) is a unsupervised multivariate technique in which new variables are
calculated as linear combinations of the old ones (metal
concentrations or percentages, pH and particle size distribution, unless otherwise stated, in our case); the new
variables, called principal components (PC), have two
main features: (i) they are uncorrelated between themselves; (ii) the rst PCs keep the main part of the variance of the original data set. In this way, it is possible to
show a great part of information by plotting the rst two
or three PCs (in this case, the rst three PCs were always
computed and the two most meaningful ones, usually

3.4.1. Total metal

Fig. 1a shows the combined plot obtained by PCA
for total metal concentrations. In general, even if no
neat grouping is present, most samples from the same
site are in the same area of the plot. S. Maurizio Canavese soil and some horizons of the other soils (e.g.
V2A and V1C) are characterised by higher metal concentrations than the other samples. It is interesting to
note that the position of the surface horizon of Poirino
soil is quite far from the ones of the lower layers, reecting a dierent composition: in fact it is poor in clay
and rich in organic matter. The V2C, V2D and V2E
samples are characterised by low concentrations of
metals, low percentages of clay and silt and high percentages of sand: this is exactly as would be expected
since sandy matrices have normally low heavy metals
sorption capacities. Besides all the horizons of Carmagnola soil are in the same area of the plot mainly

554

O. Abollino et al. / Chemosphere 49 (2002) 545557

Fig. 2. Combined plot of scores and loadings on PC1PC2 for

the percentages of the extracted metals: (a) in the rst extraction
of Tessiers procedure, and (b) in the second extraction.
Fig. 1. Total metal concentrations: (a) combined plot of scores
and loadings on PC1PC2, and (b) dendrogram obtained by
cluster analysis.

because they contain low percentages of sand. The

dendrogram, reported in Fig. 1b, conrms the dierences and similarities visible in the PCA plot, and in
particular allows us to distinguish Carmagnola and, in
smaller entity, S, Maurizio Canavese soils from the other
ones. Vigone 1, Vigone 2 and Porino do not form separate clusters.
Sand and pH are the only parameters with negative
loadings on PC1. A clear anticorrelation between sand
and silt percentages is present. The correlations among
metals, in particular for MnCu, FeNiTi, AlZn,
cannot be clearly explained in terms of chemical properties, origin or biological function.
We performed the PCA considering also CEC and
the percentage of organic matter as variables, and the
samples for which these data are available as object. In
the plot of PC1 vs. PC2 (not shown) obtained from these
data we note that organic matter is correlated to CEC,
suggesting that the binding sites on organic matter, such
as the negatively charged humic acids, contribute to the
retention of cations in soil. These parameters are anticorrelated to the percentage of clay, probably because
the soils from the investigated areas mainly contain
kaolinite, which has a low CEC. Finally, CEC is anti-

correlated or not related to metal concentrations, probably because such metals are mainly bound to the soil
matrix and are present in the exchangeable form only in
very low percentages, as shown by the results obtained
with Tessiers speciation protocol.
3.4.2. Tessiers fractions
In the plot of PC1 vs. PC2 obtained from the percentages of extracted metals in the rst fraction (Fig.
2a), it is possible to note one group formed by S.
Maurizio Canavese and Poirino soils, sample MA being
distinguished from the others because it is characterised
by a lower pH value. The samples from Carmagnola are
also close to each other. Vigone 1 and Vigone 2 samples
are partially overlapped.
Fig. 2b shows the results of PCA for the data obtained in the second step of Tessiers extraction. It is
possible to identify the following groups: (i) MB, MC
and PB, PC samples, with sample MC partially apart
because of its higher sand content; the topsoils from S.
Maurizio Canavese and Poirino, i.e. samples MA and
PA, clearly dier from the corresponding subsoils; (ii) all
horizons of Vigone 1, (iii) all horizons of Vigone 2, (iv)
all horizons of Carmagnola soil. It is evident that the
samples of the rst two groups are characterised by high
percentages of clay and low percentages of extracted Al,
Fe and Mn; the samples from Carmagnola are distinguished from the others because they have the lowest

O. Abollino et al. / Chemosphere 49 (2002) 545557

Fig. 3. Combined plot of scores and loadings on PC1PC2 for

the percentages of the extracted metals: (a) in the third extraction of Tessiers procedure, and (b) in the fth extraction.

percentage of sand and high percentages of Cd and Mn.

Sample PA is far from the others because it contains the
highest percentages of extracted Cd, Mn, Zn.
It is interesting to note also that the plots, referring to
the percentages of extracted metals in the third, fourth
and fth fractions (Figs. 3a,b, 4a), show analogous
sample groups, with few dierences: for instance, the
subsoil samples from S. Maurizio Canavese and Poirino
form two separate groups; moreover, in the fourth
fraction, V2C, V2D, V2E samples are distinguished
from V2A and V2B because of their high percentages of
cadmium and titanium. In all fractions the top horizons
of Poirino and S. Maurizio Canavese markedly dier
from the lower ones and from all other samples. Moreover it is evident that Vigone 2 samples are characterised
by low percentages of most metals in the rst four
fractions. Therefore metals result to be less available in
these soils and are mainly present in the fth fraction.
This behaviour could be associated with the high content in sand present in these samples.
Therefore the soil origin inuences the behaviour of
metals towards extraction. The groups present in PCA
plots for the ve fractions are clearly visible also from

555

Fig. 4. Percentage of the extracted metals in the fourth extraction of Tessiers procedure: (a) combined plot of scores and
loadings on PC1PC2, and (b) dendrogram obtained by cluster
analysis.

the dendrograms; as an example the dendrogram relative to the fourth fraction is reported in Fig. 4b.
PCA was also performed with all variables. As expected, CEC was found to be strongly correlated to the
percentages of Mn, Al, Fe extracted in the rst fraction,
corresponding to exchangeable metals. On the other
hand, this variable was anticorrelated to the metal percentages in the fth fraction, i.e. to the species more
strongly bound to the soil matrix.

3.4.3. Extraction with EDTA

With regard to these results, the plot of PC1 vs. PC3
(Fig. 5) is more interesting than the one of PC1 vs. PC2.
The direction of the pH and percentage of sand vectors
is opposite to the one of the vectors corresponding to
other parameters. This is an evidence that the extractability of the metals actually decreases with increasing
pH, as expected: metals are more easily released from
soils at low pHs because they are more weakly complexed by hydroxy groups and organic ligands. The rst
layers of Vigone 1 and Vigone 2 soils are characterised
by higher extraction percentages. The similarity between
these samples (V1A and V2A) is conrmed also by the
dendrogram.

556

O. Abollino et al. / Chemosphere 49 (2002) 545557

Acknowledgements
We thank the Ministero dellUniversit
a e della Ricerca Scientica e Tecnologica (MURST, Rome, COFIN
2000) and the Italian National Research Council
(C.N.R., Rome) for nancial support.

References

Fig. 5. Combined plot of scores and loadings on PC1PC3 for

the percentages of the metals extracted with EDTA in Vigone 1
and Vigone 2 soils.

The higher availability of metals in the topsoils could

reect an anthropogenic input, since exogenous metals
are usually more weakly bound to the soil matrix and
therefore more easily released (Li et al., 1995).

4. Conclusions
Total metal concentrations in the ve investigated
agricultural soils t in the typical ranges for unpolluted
soils, with the exception of lead and cadmium in some of
the sites. The concentrations in Vigone 2 samples decreased with increasing depth, whereas a surface depletion was observed for Carmagnola and Poirino.
Of course the results of this study apply to only a
limited zone of the considered areas, because of the
restricted number of samples, and cannot be directly
extended to the whole sites. The main goal of the
investigation was understanding metal mobility and
distribution also in connection with main soil characteristics.
For this reason metal speciation and mobility were
studied with Tessiers sequential extraction procedure.
The highest percentages of metals were found to be
strongly bound to soil matrix, i.e. in a form not readily
available for introduction into the food chain. Extractability in EDTA was investigated in Vigone 1 and Vigone 2 soils: in all cases metals were found to be more
readily available from topsoil than from subsoil. The
chemometric study enabled us to evidence similarities or
dierences among the ve soils and correlations or anticorrelations among variables that were not clearly visible from an examination of the analytical data in the
tables. For instance, samples from the same site showed
a similar behaviour toward extraction in Tessiers protocol, and soil pH was found to be anticorrelated to the
percentages of metals extracted by EDTA, owing to the
higher element mobility at low pHs.

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