Environ Earth Sci

DOI 10.1007/s12665-015-4096-1

ORIGINAL ARTICLE

Fractionation of heavy metals and evaluation of the environmental

risk for the alkaline soils of the Thriassio plain: a residential,
agricultural, and industrial area in Greece
Dionisios Gasparatos Georgia Mavromati
Panagiotis Kotsovilis Ioannis Massas

Received: 30 June 2014 / Accepted: 22 January 2015

Springer-Verlag Berlin Heidelberg 2015

Abstract The purpose of the present study is to test

metals accumulation and behavior in surface soils of
Thriassio plain, Attica, an area registered as probably the
most polluted in Greece. Avoiding sampling close to
obvious specific pollution sources, 50 surface soil samples
were collected and the Tessier fractionation scheme was
applied to determine the chemical partitioning of Pb, Cu,
Zn, Ni and Cr. Five chemical fractions of the studied
metals were defined: exchangeable (F1), acid-soluble (F2),
reducible (F3), oxidizable (F4) and residual (F5). The
highest Cu, Ni and Cr concentrations were measured in the
residual fraction, while the highest Zn and Pb concentrations were found in the reducible fraction. However, F3 Cr
and Ni concentrations were also high. These increased
amounts of Zn and Pb and to a lesser extent those of Cr and
Ni found in the reducible fraction indicate a potential
hazard of metals mobilization under flooding and anaerobic conditions due to excess irrigation or rain water.
Concentration of Pb in F1 was high suggesting recent
pollution episodes. Principal component analysis (PCA)
showed that the exchangeable fraction of Zn, Pb, Cr and Ni
is strongly related to the soil clay content and the oxidizable fraction of Cr and Ni to organic matter content.
According to PCA results, no other clear relation between
the extracted metal fractions and the soil components

D. Gasparatos
Department of Hydraulics, Soil Science and Agricultural
Engineering, Soil Science Laboratory, School of Agriculture,
Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece
G. Mavromati  P. Kotsovilis  I. Massas (&)
Department of Natural Resources and Agricultural Engineering,
Laboratory of Soils and Agricultural Chemistry, Agricultural
University of Athens, 75 Iera Odos Str., 11855 Athens, Greece
e-mail: massas@aua.gr

(i.e. CaCO3 eq., organic matter, clay and amorphous iron

oxides) was observed. The weak relation of CaCO3 eq.,
content with many metal fractions suggests that carbonates
affect the chemical partitioning of metals in alkaline soils
with high CaCO3 eq., content. The high values of mobility
factor for Pb, Cu, Ni and Zn point to a considerable risk
since these metals may accumulate in soil biota and plants.
Keywords Soil  Heavy metals  Sequential extraction 
Mobility factor  Principal component analysis

Introduction
The potentially harmful substances emitted into the environment due to anthropogenic activities, that can also
adversely affect ecosystems and human health, have been a
principal concern of the scientific community during the
last decades. Contamination of soils with heavy metals can
be harmful to the biota and humans, and thorough research
has been carried out recently in many countries and regions
(Gasparatos 2013).
Soils in urban and industrial areas are known to have
atypical characteristics such as unpredictable layering, poor
structure, and high concentrations of trace elements (Ajmone-Marsan and Biasioli 2010; Massas et al. 2010). These
soils differ greatly from natural soils as they are greatly
influenced by anthropogenic activities, and are the recipients of large amounts of heavy metals from a variety of
sources including industrial wastes, vehicle emissions, coal
burning waste, accumulation of garbage and other solid
wastes, effluents from households, and from various amenities like dry cleaners. Enrichment of urban and industrial
zone soils with metals is a limiting factor for the sustainability of soil ecosystems, the quality of life of town and city

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Environ Earth Sci

residents, and for the improvement of working environments (Luo et al. 2012; Argyraki and Kelepertzis 2014).
The accumulation of heavy metals in soils and sediments
causes a potential risk to humans because these environmental media can act as transmitters of various contaminants as metals in surface-ground water, atmosphere and
living organisms (Massas et al. 2013). The toxicity and
mobility of heavy metals in soils depend not only on their
concentrations, but also on their chemical associations and
properties, and on soil properties such as pH, redox potential, biological root activity and the formation of chelates.
For these reasons it is essential to identify the speciation of
metals in different soil fractions for the determination of
their potential mobility, availability and persistence in the
environment (Dousis et al. 2013).
Sequential extraction procedures have been widely
applied to soils and aquatic sediments to characterize
chemical phase associations of metals. These procedures
are probably the most functional tool for predicting longterm behavior of metals in contaminated solid materials
and have been extensively used in scientific research due to
the toxicity of heavy metals (Zimmerman and Weindorf
2010). A sequential extraction scheme provides information for the differentiation of the relative bonding strength
of metals in various solid phases and for the potential
reactivity of metals under different physicochemical environmental conditions; in that sense such schemes are
considered useful for evaluating mobility and bioavailability of heavy metals in metal enriched soils (Lu et al.
2003). Among the various sequential extraction protocols
the one proposed by Tessier et al. (1979) is most often
followed.
The present study is a follow up to a recently published
work (Massas et al. 2013), and indents to examine in depth
metals accumulation and behavior in surface soils of an
area considered as one of the most polluted in Greece
(Thriassio plain, Attica). The main objectives are: (a) to
determine the chemical partitioning of Cu, Zn, Pb, Ni and
Cr applying the Tessier sequential extraction scheme in
surface soil samples and to assess the amounts and
potential mobility of metals in these soils and (b) to test the
relationships between various fractions of the above metals
and the soil components expected to control metal mobility
by using principal component analysis (PCA) and cluster
analysis.

Materials and methods

Study area
Thriassio plain is located 25 km north-west from the center
of Athens and is separated into four municipalities, Elefsis

123

(26,000), Aspropyrgos (28,000), Mandra (13,000), and Nea

Peramos (7,000), and the community of Magoula (4,000)
(population in parentheses; 2001 census). The geology of
the plain consists of alluvial and diluvial formations of
Quaternary deposits and the topography is mostly flat, with
a gentle slope towards the sea not exceeding 3 %. Recent
data report at least 2,200 large and small industrial plants
operating in the area, such as steel, steelwork, chemical,
product, non-metal mineral material and cement, as well as
oil refineries (Mavrakis et al. 2008). The various economic
activities have been chaotically developed, under a practically absent central control and planning context. Residential, agricultural, industrial and harbour areas are
intermixed with no distinct boundaries (Lykoudis et al.
2008; Massas et al. 2013). Air pollution in the Thriassio
plain mainly originates from industrial activities, while in
the adjacent Athens city, it is primarily from traffic emissions (Mavrakis et al. 2005). Thriassio plain probably
exhibits the highest (a) industrial activity concentration,
(b) fuel consumption, and (c) pollution related to the production processes, in all of Greece (Lykoudis et al. 2008).
The nearby existence of Athens landfill imposes an additional environmental threat to the area. The plain is also
crossed by highways, avenues and secondary roads. The
climate in the Thriassio plain is typically Mediterranean.
The mean annual rainfall is 390 mm; the average temperature is 18 C and north winds are prevailing.
Soil sampling
Due to the existence of the military airport and two main
highways, the studied area was divided into five sampling
sections (Fig. 1). To demonstrate the mobility and bioavailability of heavy metals in the soils of the Thriassio
plain in a manner representative of their distribution pattern
in the area, the selected sampling sites were located far
from noticeable site specific pollution sources. In each one
of the sampling areas a grid sampling scheme (cell size
250 m 9 250 m) was followed and a total of 50 top soil
samples (05 cm depth) were collected (Fig. 1). At all
sampling sites three sub-samples, each from a 40 9 40 cm
surface area, were obtained and mixed to make a composite
sample.
Analytical methods
The soil samples were air-dried and sieved through a 2-mm
sieve. The organic matter content and the mechanical
composition of the samples were determined by the
WalkleyBlack procedure (Nelson and Sommers 1982) and
by the Bouyoucos hydrometer method (Bouyoucos 1951),
respectively. The soil samples were also tested for their pH
values by the use of standard glass/calomel electrodes in

Environ Earth Sci

Fig. 1 Study area and sampling
sites

1:1 w/v soilwater ratio slurries and for the CaCO3

equivalent content, by measuring the evolved CO2 following HCl dissolution. Determination of amorphous and
poorly crystallized iron oxides was achieved by using
0.04 M hydroxylamine hydrochloride in 25 % acetic acid
(pH 2) at high temperature (95 C) (Gleyzes et al. 2002).
To obtain the pseudo-total metal concentrations the
soil samples were digested with aqua regia (Gasparatos and
Haidouti 2001). The term pseudo-total is applied
because aqua regia digestion does not completely destroy
silicates (Facchinelli et al. 2001). The bioavailable fraction
of the studied metals was extracted by DTPA (diethylene
triamine pentaacetic acid) (Lindsay and Norvell 1978). In
the text, the terms total and available are used for the
aqua regia and DTPA extracted metal fractions,
respectively.
The sequential extraction proposed by Tessier et al.
(1979), was followed to examine the chemical fractions of
Cu, Zn, Pb, Ni and Cr in the studied soils. Each fraction
was defined as follows:
Exchangeable (fraction 1F1): 2.00 g of soil extracted
with 16 ml 1 M MgCl (pH 7) in 100 ml polyethylene
centrifuge tubes for 1 h at room temperature with continuous agitation.

Acid-soluble or bound to carbonates (fraction 2F2):

residue from exchangeable fraction, extracted with 16 ml
1 M CH3COONa (pH 5) for 5 h at room temperature with
continuous agitation.
Reducible or bound to FeMn oxides (fraction 3F3):
residue from acid-soluble fraction, extracted with 40 ml of
0.04 M NH2OH.HCl in 25 % acetic acid (v/v) for 5 h at
96 3 under intermittent agitation.
Oxidizable or bound to organic matter (fraction 4F4):
residue from reducible fraction, extracted with 6 ml of
0.02 M HNO3 and 10 ml of 30 % H2O2 adjusted with pH 2
with nitric acid. The mixture was heated to 85 2 for 2 h
with occasional agitation. After cooling, 10 ml of 3.2 M
CH3COONH4 in 20 % (v/v) HNO3 was added and the
sample diluted to 40 ml and agitated continuously for
30 min.
Residual (fraction 5F5): the residual fraction was
determined as the difference between the total metal concentration and the sum of the contents in the four previous
fractions.
After each extraction the suspension was subjected to
centrifugation for 20 min at 4,000 rpm. The supernatant
was removed with a pipette; care was taken not to remove
any of the solid residues. All metal concentrations were

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Environ Earth Sci

determined by atomic absorption spectrophotometry, using

a Varian-spectra A300 system. At all stages of sample
preparation and analysis stringent precautions were taken
to minimize contamination through air, glassware and
reagents. A control sample was analyzed for every 10
samples and reproducibility was tested by reanalyzing
30 % of the samples. The analytical precision, estimated as
relative standard deviation, was less than 3 %.
ERM-CC141 loam soil was used as reference material to
check the reliability of the followed aqua regia procedure.
The mean recovery of metals was 96, 98, 104, 105 and
94 % for Cu, Zn, Ni, Pb and Cr, respectively.
The descriptive statistical parameters, correlation analysis, principal component analysis and cluster analysis
were carried out using the statistical software STATISTICA for Windows (StatSoft, Inc., USA, 1995, Version 7).

Results
Soil properties and heavy metal concentrations
The main soil properties are presented in Table 1. The soil
samples are characterized by alkaline pH values ranging
from 7.0 to 8.5 with a mean of 7.9. Alkaline pH values are
attributed to the presence of carbonates whose content
ranges between 37 and 730 g kg-1 with a mean of
252 g kg-1. The high content of carbonates in the study
area is mainly derived from the calcareous nature of
Quaternary deposits which have filled the basin of
Thriassio plain. Soil particle size distribution showed that
the clay content averaged 251 g kg-1 and the majority of
the studied soils have light to medium texture. The organic
matter content is greatly varied between 0.7 and
110 g kg-1 with a mean value of 33 g kg-1. Amorphous
iron oxides concentration, the most active fraction of free
iron oxides, was low and averaged 1.1 g kg-1 with a range
of 0.63.1 g kg-1. According to Table 2, the concentrations of total and available metal forms fell into the ranges
reported recently for the study area and are not considered
as particularly high (Massas et al. 2013).

Fractionation of heavy metals

The mean values of metal fraction percentages and associated tables of metal fraction percentages descriptive statistics and ShapiroWilk test p values are shown in Fig. 2.
Copper
The distribution pattern of Cu indicates that this metal is
mainly associated with the residual fraction (42 %) followed
by the oxidizable fraction (24.5 %). The high percentage of
Cu in the residual fraction has been also reported by other
researchers (Moral et al. 2005; Iwegbue 2013). The considerable amounts of Cu extracted in the fourth step of the
fractionation process from some soil samples, indicate the
high stability of Cu-organic complexes and the tendency of
this element to associate with the soil organic matter (Papafilippaki et al. 2007). The percentages of Cu in the reducible
and acid-soluble fraction were almost equal *15 % while
the exchangeable fraction accounts for less than 4 %. The
average Cu contents associated with different fractions were
in the following order: F5 [ F4 [ F2 * F3 [ F1, suggesting low solubility and mobility of Cu at the existing soil
physicochemical conditions.
Zinc
The partitioning scheme of Zn shows that this metal is
strongly associated with the reducible fraction (43.4 %).
Kabala and Singh (2001) concluded that Zn sorption on
FeMn oxides is probably the most important mechanism
that controls the behavior of this element in the soils. Our
findings are comparable to the results obtained recently by
Jiang et al. (2013) who found that the reducible fraction
represents the 49.5 % of the total Zn. The tendency of clay
minerals to absorb zinc irreversibly by lattice penetration
may explain the significant amounts of Zn found to the
residual fraction. The acid-soluble fraction of Zn was in the
range 4.821.2 % (mean *12 %). This can be attributed to
carbonate minerals that in soils form double salts complexes with Zn (Iwegbue 2013). The presence of Zn in the

Table 1 Descriptive statistics of selected physicochemical properties of the studied soils (N = 50)
Clay (g kg-1)

Silt (g kg-1)

Sand (g kg-1)

Org. Mat. (g kg-1)

CaCO3 eq., (g kg-1)

Amorphous Fe2O3 (g kg-1)

pH (1:1)

Mean

251

288

469

33

252

1.1

7.9

Median

239

295

477

31

246

1.0

8.0

Min.

103

94

46

0.7

37

0.6

7.0

Max.

508

446

733

112

730

3.1

8.5

SD

81

68

131

22

131

0.4

0.4

C.V. %

33

23

28

66

52

38

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Environ Earth Sci

Table 2 Descriptive statistics
for the total and available
concentrations of the studied
metals in the soils of Thriassio
plain (N = 50)

Cr (mg kg-1)

Zn (mg kg-1)

Ni (mg kg-1)

Pb (mg kg-1)

Cu (mg kg-1)

Mean

89.3

160.4

102.8

120

42.0

Median

94.3

155.2

97.8

121.2

37.7

Min.

41.4

73.0

41.4

67.5

15.4

Max.

148.7

543.5

175.0

215.8

318.2

SD

26.7

72.5

38.4

135.5

41.3

C.V. %

37

45

37

23

98

Mean

0.8

5.3

2.1

7.1

3.1

Median

0.7

4.5

2.3

5.6

1.9

Min.

0.1

0.9

0.5

2.0

0.5

Max.

2.6

25.6

3.6

33.9

59.9

SD

0.7

4.3

0.7

5.2

8.5

C.V. %

82

81

39

73

263

Total

Available

oxidizable and especially in the exchangeable fraction is

negligible. The low levels of exchangeable Zn (*1.5 %)
indicate the low solubility of Zn in the studied calcareous
soils. The percentage of Zn associated with different
fractions was in the order F3 [ F5 [ F2 [ F4 [ F1.
Lead
Lead mainly found in the reducible (36.7 %) and in the
residual (26.2 %) fraction. The acid-soluble (F2) and the
oxidizable fraction account for about 16 and 8.8 % of total
lead, respectively. However, the exchangeable Pb (F1)
ranged from 6.2 to 22 % (mean 12.3 %) implying that the
mobility and bioavailability of Pb were the highest among
the metals. Many studies have demonstrated the strong
affinity of Pb for both Fe and Mn compounds (Sauve et al.
2000; Businelli et al. 2003). Fe and Mn compounds sorb
Pb2? via formation of inner sphere sorption complexes
(Scheinost et al. 2001). In soils with low organic matter as
the studied soils, this process seems to be the major
mechanism controlling the fate and transport of Pb into the
aquatic environments and the biota. Indeed, a relatively
small fraction of Pb is bound to organic matter and the
highest percentages (*90 %) are associated with mineral
matrix of the soil (Fig. 2). The association of Pb with the
exchangeable fraction indicates dominance of anthropogenic sources through emissions from industry, vehicles,
and/or agricultural activities. The percentage of Pb associated with the different fractions was in the order:
F3 [ F5 [ F2 [ F1 [ F4.
Nickel
The dominant fraction of Ni was the residual (43.8 %),
indicating that the presence of Ni in the studied soils is

mainly controlled by the soils parent material. Many

studies showed that Ni in soils mostly occurred in the
residual fraction ([50 %) (Ryan et al. 2002; Iwegbue et al.
2007). The second most common fraction was the reducible suggesting that FeMn oxides are another important
pool of Ni in the studied soils, probably due to Ni (II)
adsorption on the negatively charged surface of Mn oxides
(specific adsorption). The oxidizable and the acid-soluble
fractions contain almost equal amounts (13.3 and 11.3 %,
respectively) while the exchangeable fraction shows the
lowest percentage of Ni (5.2 %). The distribution pattern of
Ni in this study is similar to the results reported by Iwegbue
(2007) who found that the exchangeable fraction was less
than 9 % of total soil Ni. The percentage of Ni associated
with the different fractions was in the order: F5 [ F3 [
F4 [ F2 [ F1.
Chromium
From the studied metals, Cr showed the highest affinity for
the residual phase with an average of 70 % of total Cr
concentration to strongly relate to soil components that
reduce metals availability. Similar results also reported by
Iwegbue (2013) and Moral et al. (2005). Chromium speciation in soil environment is mainly affected by the
organic matter content, the hydrous metal oxides concentration, the redox potential and the pH value. In this study
equal amounts of Cr were extracted under reducing and
oxidizing conditions (mean values 12.8 and 12.5 %,
respectively) supporting the significant role of FeMn
oxides and organic matter in Cr dynamics in the soil system. Very low (\3 %) chromium quantity was found in
exchangeable and in acid-soluble fractions indicating the
low solubility of Cr in the soils of Thriassio plain. The
percentage of total Cr concentration associated with the

123

35

35
30

MeanSE

25
20
15

Cu-F1%

Cu-F2%

Cu-F3%

Cu-F4%

5
0

Cu-F5%

Pb-F1%

Metal
Fraction
(%)

Mean

Median

Min.

Max.

10th
percentile

90th
percentile

Std.
Deviation

F1
F2
F3
F4
F5

5,23
14,53
13,53
24,50
42,21

5,15
12,57
12,14
21,85
45,44

1,89
5,38
3,17
5,90
8,66

9,58
34,43
35,14
62,75
70,86

3,02
8,40
6,35
9,86
15,18

7,56
24,41
22,23
41,23
60,28

1,90
6,32
6,81
13,09
16,38

Shapiro

Wilk
test
p value*
0,18
0,00
0,00
0,00
0,12

* p > 0,05 indicates normal distribution

Mean0,95 Conf. Interval

45

35

Pb-F4%

Pb-F5%

Median

Min.

Max.

10th
percentile

90th
percentile

Std.
Deviation

F1
F2
F3
F4
F5

12,34
15,98
36,75
8,77
26,16

11,70
14,47
36,02
8,17
26,97

6,23
8,64
19,95
4,40
15,05

22,01
32,01
58,31
16,61
35,62

7,14
12,48
27,77
5,59
18,01

18,09
23,08
46,66
12,59
31,61

4,12
4,53
7,94
2,95
4,69

40

Shapiro

Wilk
test
p value*
0,06
0,00
0,58
0,02
0,02

(%)

30

25
20
15

20

10
Mean

Mean

10
5

0
Ni-F1%

Ni-F2%

NiF3%

Ni-F4%

Ni-F5%

th

th

Metal
Fraction
(%)

Mean

Median

Min.

Max.

10
percentile

90
percentile

Std.
Deviation

F1
F2
F3
F4
F5

5,24
11,32
26,29
13,35
43,81

4,20
10.03
25,56
12,39
45,73

2,15
5,57
13,06
4,36
10,93

11,45
23,07
46,72
30,04
74,47

2,81
6,75
19,28
5,21
26,83

9,47
19,03
36,12
21,01
59,44

2,51
4,53
6,27
6,23
14,33

Shapiro

Wilk
test
p value*
0,00
0,00
0,13
0,02
0,41

Mean0,95 Conf. Interval

80
70

50

MeanSE

40
30
20

Mean

10
Cr-F1%

Cr-F2%

Cr-F3%

Zn-F1%

Zn-F2%

Zn-F3%

Cr-F4%

Cr-F5%

Metal
Fraction
(%)

Mean

Median

Min.

Max.

10
percentile

90th
percentile

Std.
Deviation

F1
F2
F3
F4
F5

2,71
1,77
12,85
12,50
70,17

2,61
1,54
11,19
12,18
72,10

1,18
0,53
6,27
0,00
45,87

4,35
4,98
29,44
22,31
86,61

1,67
0,87
7,73
7,63
57,88

3,85
2,94
20,65
19,15
80,18

0,83
0,88
5,15
4,88
8,47

* p > 0,05 indicates normal distribution

th

Shapiro

Wilk
test
p value*
0,04
0,00
0,00
0,19
0,21

Zn-F4%
th

Zn-F5%

Metal
Fraction
(%)

Mean

Median

Min.

Max.

10
percentile

90th
percentile

Std.
Deviation

F1
F2
F3
F4
F5

1,54
11,97
43,39
3,28
39,82

1,57
12,00
44,33
2,59
39,45

0,35
4,84
14,27
1,18
4,93

3,62
21,28
76,15
14,16
76,75

0,84
8,09
26,30
1,80
21,05

2,31
15,11
59,40
5,30
60,68

0,60
3,18
13,52
2,28
16,38

* p > 0,05 indicates normal distribution

* p > 0,05 indicates normal distribution

(%)

Pb-F3%

Mean

50

MeanSE

(%)

30

123

Pb-F2%

Metal
Fraction
(%)

* p > 0,05 indicates normal distribution

50

Mean

Mean

60

20

10

25

15

10

40

Mean0,95 Conf. Interval

(%)

30

40

MeanSE

40

45

MeanSE

45

(%)

50

Mean0,95 Conf. Interval

Mean0,95 Conf. Interval

Environ Earth Sci

Shapiro

Wilk
test
p value*
0,02
0,25
0,80
0,00
0,77

Environ Earth Sci

b Fig. 2 Mean values of metal fraction percentages and associated

tables of metal fraction percentages descriptive statistics and Shapiro

Wilk test p values to check the hypothesis of normal distribution;
a Cu, b Pb, c Ni, d Zn, e Cr; F1 exchangeable fraction, F2 acidsoluble fraction, F3 reducible fraction, F4 oxidizable fraction, F5
residual fraction (N = 50)

different fractions followed the order: F5 [ F3 * F4 [

F2 [ F1.

Discussion
Metal fractions
Metal additions to soils due to either anthropogenic activities or physical phenomena lead progressively to more
stable forms of metals in the soil matrix and under no metal
additions metal fractions distribution in mineral soils is
expected to follow the order F1 \ F2 \ F3 \ F4 \ F5
(Kabala and Singh 2001). Indeed, this is observed for Cu
and Cr, while for Pb, Zn and Ni a different pattern was
observed; the reducible fraction percentage of the latter
metals is higher than that of the oxidizable phase. For Pb
the reducible fraction percentage is higher even than the
residual fraction percentage (Fig. 2). Jiang et al. (2013)
also found that Pb was mainly associated with the reducible
fraction in heavily polluted soils from Qingshuitang District, a typical heavy industrial base especially in smelting
and chemicals in China. Considering the land uses in
Thriassio plain, such observation points to that the additions of Pb, Ni and Zn in the soils of the area is an ongoing
procedure due to emissions from anthropogenic activities.
As the sum of the three first fractions (F1 ? F2 ? F3)
accounts for the 65, 43 and 57 % of the total Pb, Ni and Zn
concentrations in the soils, it is highly possible that under
modified soil conditions the solubility and availability of
these metals can increase, with profound implications for
the soil environment, the underground waters and the food
chain. In addition, the reducible fraction percentage for Pb,
Ni and Zn represents the 37, 26 and 43 %, respectively, of
the total concentrations posing an environmental risk since
excess irrigation water application or heavy rain may result
to flooding phenomena and to limited or no oxygen concentration in soil pores, especially at micro sites in soil
aggregates, and to reducing conditions thus increasing the
availability of these metals (Borch et al. 2010). The
90th percentile and the maximum values for the reducible
fraction percentage, suggest that under adverse soil conditions such situation is more likely to occur for Zn (Table
in Fig. 2).
According to the ShapiroWilk test p values (Fig. 2 and
associated tables) the Cu and Pb exchangeable fraction

percentage values follow normal distribution indicating a

tendency for the values to gather close to mean. On the
contrary, the respective values for Zn, Ni and Cr are not
normally distributed around the mean probably due to soil
characteristics or to factors that might be of anthropogenic
origin as recent site specific pollution events. Except Pb,
the residual fraction percentage values for the tested metals
are normally distributed around the mean, pointing to the
fact that the stability of metal forms associating with the
residual fraction is not significantly influenced by other
factors than their original incorporation in the soil parent
material as a result of the soil formation processes.
Regarding normal distribution of the other metal fraction
percentage values, mixed trends were observed depending
on the metal-fraction combination, indicating a different
transformation rate of metal forms governed by soil and
metal properties as well as by soil enrichment with metals.
Principal component analysis and cluster analysis
Principal component analysis (PCA) was performed on
metal fractions of 50 soil samples, to reduce the variables
to only two principal factors which can be used to interpret
the chemical affinity of metal factions for soil components.
Figure 3 shows that cumulative variances of the Cu, Zn,
Pb, Cr and Ni bound to soil components (clay, carbonates,
FeMn oxides and OM) can be described by factor 1 and
factor 2 which respectively accounted for 84.8, 54.8, 58.8
and 72.7 of the total variance.
As it is apparent from Fig. 3, no clear relation of soil
components with the metal fractions exists, except two
distinct groups, the one of clay content with the
exchangeable fraction of Zn, Pb, Ni and Cr and the other of
organic matter content with the oxidizable fraction of Cr
and Ni. This grouping indicates that the availability of Zn,
Pb, Ni and Cr is mainly controlled by the inorganic colloids
chemistry in soils with low organic matter content and that
the Cr- and Ni-organic complexes are highly stable. The
exchangeable fraction of metals (except Cu) also positively
correlate to the clay percentage (p \ 0.001) further supporting that the exchangeable metals phase is primary
associated with clay minerals.
However, the joint clustering of Cu, Zn and Pb under
reducing and oxidizing ambient conditions that is independent from the amorphous Fe oxides and the soil organic
matter content, in addition to Zn and Cu clustering under
acidifying conditions that is independent from carbonates
concentration, suggest common origin of these so called
urban metals and possibly similar chemical transformation processes of these metals in the soils of the studied area.
Attempting to better clarify the relation of metal fractions to the soil components, a cluster analysis was applied
to the data set. As it is presented in Fig. 4, clay content is

123

Environ Earth Sci

1,0

1,0
Cu1

0,5

Zn1
Pb1

0,0

a
Factor 2 : 22,85%

Factor 2 : 15,40%

Fig. 3 Principal component

analysis for Zn, Cu, Pb, Cr and
Ni chemical fractions and
properties of Thriassio plain
soils; a exchangeable fraction
and clay content, b acid-soluble
fraction and CaCO3 eq.,
content, c reducible fraction and
amorphous oxides content,
d oxidizable fraction and
organic matter content (N = 50)

Cr1
Ni1

CLAY

-0,5

Cu3

0,5

Fe2O3
Zn3
Pb3

0,0

Cr3

-0,5

Ni3

-1,0

-1,0
-1,0

-0,5

0,0

0,5

-1,0

1,0

-0,5

Factor 1 : 69,38%

0,0

0,5

1,0

Factor 1 : 35,93%
1,0

1,0

Ni4 Org Matter

0,5

Pb2

Factor 2 : 33,85%

Factor 2 : 25,42%

Cr4

0,0
Ni2

Cr2

Cu2

Pb4

0,0

-0,5

-0,5
CaCO3

Zn2

-1,0

-1,0
-1,0

-0,5

0,0

0,5

Factor 1 : 29,36%

strongly associated with the exchangeable fraction of Zn

and Ni and less powerfully with the exchangeable fraction
of Pb and Cr. Clay and organic matter weakly related to the
residual fraction of all metals, and amorphous iron oxides
appear to be the less active soil component in the studied
soils.
Cluster analysis revealed a connection of CaCO3 eq.,
content with the reducible fraction of Cr and a weak
association of CaCO3 eq., content with the reducible and
oxidizable fractions of Zn and Pb. In addition, CaCO3 eq.,
weakly associates with all Cu fractions except the residual.
It seems that the high carbonate salts concentration (mainly
CaCO3) affects and to some extent controls the solubility
of metals in the studied soils. These results also imply
strong antagonism among the various soil components for
heavy metals binding, as previously suggested by Yu et al.
(2001) for river sediments in Taiwan.
Mobility factor
The mobility of heavy metals in soils may be evaluated on
the basis of their potential bioavailable or active forms as
determined by sequential extraction scheme. A mobility

123

Zn4
Cu4

0,5

1,0

-1,0

-0,5

0,0

0,5

1,0

Factor 1 : 38,34%

index which is also called mobility factor was computed on

the basis of the ratio of exchangeable (F1) and acid-soluble
(F2) fractions to the sum of all fractions (Iwegbue 2013)
(Eq. 1).
MF

F1 F2
 100
F1 F2 F3 F4 F5

A high MF value indicates that metals can potentially

enter to the biota or they can move downwards in the soil
profile contaminating the ground waters, thus posing a
considerable threat to the local environment or even to
remote ecosystems. The descriptive statistics of computed
mobility factors are displayed in Table 3.
Based on the calculation of MFs, the mobility of metals
in this study follows the order: Pb [ Cu [ Ni [ Zn [ Cr.
The indices of mobility were comparatively highest for
Pb (mean value 28.3 %) while Cr had the lowest MF (mean
value 4.5 %) showing the high stability of Cr in the specific
soil conditions. However, the MF values of Pb, Cu, Ni and
Zn are higher than 10 %, suggesting that their availability
points to serious soil enrichment and to increased risk since
these metals may accumulate in soil micro-organisms and
plants (Lei et al. 2010).

Environ Earth Sci

Fig. 4 Hierarchical clustering
results (dendrogram) for Zn, Cu,
Pb, Cr and Ni chemical
fractions and clay, CaCO3 eq.,
amorphous iron oxides and
organic matter content in the
soils of Thriassio plain
(N = 50)

Weighted pair-group average

1-Pearson r
Cu1
Cu2
Cu3
Cu4
Zn4
Zn2
Zn3
Pb3
Pb4
Cr3
CaCO3
CuR
CrR
NiR
Zn1
Ni1
CLAY
Pb1
Cr1
Ni3
Ni2
Cr4
Ni4
ZnR
PbR
Org Matter
Pb2
Fe2O3
Cr2
0,0

0,2

0,4

0,6

0,8

1,0

1,2

Linkage Distance

Table 3 Descriptive statistics for mobility factors of the studied

metals in the soils of Thriassio plain (N = 50)
Mobility factor
Cr (%)

Zn (%)

Ni (%)

Pb (%)

Cu (%)

Mean

4.5

13.5

16.5

28.3

19.8

Median

4.0

13.2

13.9

27.8

17.7

Min.
Max.

2.3
8.9

7.2
22.3

8.1
32.0

16.6
43.7

7.2
42.0

SD

1.4

3.1

6.7

5.7

7.5

C.V.

38

23

41

20

38

For the soils of the area, the order of availability of the

studied metals as indicated by the mean MF values is similar
to that suggested by the availability ratios (ARs) (i.e. the
DTPA extracted metal fraction expressed as percentage of
the aqua regia extracted fraction). Indeed, Massas et al.
(2013), found very low availability ratios (ARs) for Cr and
high ARs for Pb and Cu. Though the chemical properties of
the extracting solutions differ, the less laborious production
of ARs and the adequate number of tested soil samples
support the AR as a promising index to estimate the availability and the potential mobility of metals in alkaline soils.

Conclusions
Though the soils of the studied area are enriched by the
tested metals, in few sampling sites the total and the

available Pb, Zn, Cu, Cr and Ni concentrations are considered as high. Sequential extraction results showed a
different equilibrium state of the studied metals pointing to
different metal enrichment of soils or to different metals
origin. Under reducing soil conditions the availability of Zn
and Pb is expected to increase promoting the incorporation
of these metals in soil biota and their circulation in soil
environment thus enhancing the risk for ground water
enrichment and for higher plant uptake. The mobility factor
for Pb, Zn, Cu and Cr was higher than 10 % and suggests
high availability of these metals. Principal component
analysis and cluster analysis results indicate that in alkaline
soils rich in carbonates most of the defined chemical metal
fractions are not closely related to the soil components that
were expected, and that in the low organic matter content
soils of the area the clay content controls the availability of
Zn, Pb, Ni and Cr. A common origin of Cu, Zn and Pb in
the studied soils is supported by the joint clustering of the
Cu, Zn and Pb reducible and oxidizable fractions and by
the joint clustering of Cu and Zn acid-soluble fractions.

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