Environmental Pollution 135 (2005) 515522

www.elsevier.com/locate/envpol

Soil remediation: humic acids as natural surfactants

in the washings of highly contaminated soils
Pellegrino Contea,b,), Anna Agrettoa,b, Riccardo Spaccinia,b,
Alessandro Piccoloa,b
a

Dipartimento di Scienze del Suolo, della Pianta e dellAmbiente, Universita` di Napoli Federico II,
Via Universita` 100, 80055 Portici, Italy
b
INCA Research Unit Napoli 2, Naples, Italy
Received 21 June 2004; accepted 15 October 2004

Solutions of natural humic acids appear to be a better choice for washing highly polluted soils.
Abstract
The remediation of the highly contaminated site around the former chemical plant of ACNA (near Savona) in Northern Italy is
a top priority in Italy. The aim of the present work was to contribute in nding innovative and environmental-friendly technology to
remediate soils from the ACNA contaminated site. Two soils sampled from the ACNA site (A and B), diering in texture and
amount and type of organic contaminants, were subjected to soil washings by comparing the removal eciency of water, two
synthetic surfactants, sodium dodecylsulphate (SDS) and Triton X-100 (TX100), and a solution of a natural surfactant, a humic acid
(HA) at its critical micelle concentration (CMC). The extraction of pollutants by sonication and soxhlet was conducted before and
after the soil washings. Soil A was richer in polycyclic aromatic hydrocarbons, whereas soil B had a larger content of thiophenes.
Sonication resulted more analytically ecient in the ne-textured soil B. The coarse-textured soil A was extracted with a general
equal eciency also by soxhlet. Clean-up by water was unable to exhaustively remove contaminants from the two soils, whereas all
the organic surfactants revealed very similar eciencies (up to 90%) in the removal of the contaminants from the soils. Hence, the
use of solutions of natural HAs appears as a better choice for soil washings of highly polluted soils due to their additional capacity
to promote microbial activity, in contrast to synthetic surfactants, for a further natural attenuation in washed soils.
2004 Elsevier Ltd. All rights reserved.
Keywords: Soil-remediation; Soil-washing; Soxhlet; Sonication; Contaminated soils

1. Introduction
Soil is a complex matrix permanently interacting with
other environmental compartments such as waters and
air (Schwarzenbach et al., 2003) and thus its pollution
can directly propagate contamination to surface and
) Corresponding author. Dipartimento di Scienze del Suolo, della
Pianta e dellAmbiente, Universita` di Napoli Federico II, Via Universita`
100, 80055 Portici, Italy. Tel.: C39 8 1253 9170; fax: C39 8 1253 9186.
E-mail address: pellegrino.conte@unina.it (P. Conte).
0269-7491/$ - see front matter 2004 Elsevier Ltd. All rights reserved.
doi:10.1016/j.envpol.2004.10.006

ground waters, and air (Fent, 2003). Soil pollution may

be due to industrial accidents (spills, leaks, and leaking
underground storage tanks) (Ballarin-Denti et al., 1999;
Lee et al., 2002; Kiem et al., 2003) and anthropogenic
activities (forest and agricultural res, fossil fuel combustions) (Simcik et al., 1999; Venkataraman et al.,
2002), and represents a long-term source of environmental contamination. Weak interactions bind soil
constituents to contaminants (Piccolo et al., 1996; Chin
et al., 1997; Zhou and Rowland, 1997; Kopinke et al.,
2000; Peuravuori, 2001; Peuravuori et al., 2002; Paaso

516

P. Conte et al. / Environmental Pollution 135 (2005) 515522

et al., 2002) which, once displaced from the weak bonds,

may be again released in the environmental compartments (Gaboriau and Saada, 2001; Huang et al., 2003;
Ockenden et al., 2003) and aect the human food-chain
(Ballarin-Denti et al., 1999; Mackay and Fraser, 2000;
Borga et al., 2001).
New soil remediation technologies emphasizes the
transformation and detoxication of pollutants. For
example, bioremediation enables permanent elimination
of pollutants by in situ remediation at low cost. However, bioremediation is limited by the correct selection of
active microbes, the appropriate soil conditions for
microbial activity, recalcitrance of pollutants to biodegradation, and formation of metabolites which may
be more toxic than the parent contaminant (Boopathy,
2000).
Among other techniques, soil washing may be a valid
and relatively inexpensive alternative in soil remediation
(Kuhlman and Greeneld, 1999). It is based on the
desorption of pollutants from contaminated soils
through the action of simple water (Griths, 1995), or
organic surfactants (Deshpande et al., 1999; Chu and
Chan, 2003) which may be non-ionic (Garon et al., 2002;
Mata-Sandoval et al., 2002) or anionic (Chun et al.,
2002). The liquid phases containing the desorbed
pollutants are then disposed or subjected to further
chemical or biochemical treatments for complete detoxication (Mann, 1999).
Dierent classes of surfactants are employed for soil
washing depending on the nature of contaminants to be
removed. For example, pesticides are removed by nonionic surfactants such as Triton X-100, and biosurfactants like the rhamnolipids (Noordman et al.,
2000; Mata-Sandoval et al., 2002). Polycyclic aromatic
hydrocarbons (PAH) can be washed away by alkylphenol ethoxylate (Garon et al., 2002), hydroxypropyl-bcyclodextrins (Cuypers et al., 2002), and other anionic
surfactants such as sodium dodecylbenzene sulphonate,
monoalkylated disulphonated diphenyl oxide, and
dialkylated disulphonated diphenyl oxide (Chun et al.,
2002). In general, water-soluble surfactants may disperse
solids and liquids immiscible in water, while surfactants
forming microemulsion solubilize hydrocarbons, and
oil-soluble surfactants facilitate water recycling (Kuhlman and Greeneld, 1999). However, the use of
surfactants in cleaning-up contaminated sites may be
limited by soil texture and their intrinsic biological
toxicity. Soils with large content of ne-sized textural
particles (silt and clay) reduce considerably washing
eciency of surfactants (Kuhlman and Greeneld, 1999;
Lee et al., 2002). The reported toxicity of surfactants on
several soil microbial cells may prevent further biodegradation of pollutants and balanced biological
activity in the washed soils (Sandbacka et al., 2000).
Humic substances (HAs), naturally occurring surfactants, begin to be recognized as a possible aid in

soil bioremediation techniques. The bioavailability of

polychlorinated biphenyls (Fava and Piccolo, 2002), and
PAH (Holman et al., 2002) appeared to be increased by
addition of exogenous humic substances to contaminated soils. Furthermore, the surfactant activity of HAs
was found to reduce sorption of organic contaminants
on spiked soils, thereby enabling desorption-remediation of PAH (Janzen et al., 1996; Chefetz et al., 2000;
Conte et al., 2001; Holman et al., 2002), dioxins (Kim
and Lee, 2002), and heavy metals (Weng et al., 2002;
Halim et al., 2003). However, no studies have been so
far reported on the application of HAs as surfactants in
the soil washing of a real and highly contaminated site.
The aim of this work was to compare the eciency of
a humic acid (HA) with that of common surfactants
(sodium dodecylsulphate, and Triton X-100) and water
in the washing of two highly polluted soils sampled in
the contaminated industrial area of a chemical plant.
The soils were dierent in texture and composition of
pollutants. One was rich in PAH, whereas the other had
a high content of thiophenes. Moreover, the eciency of
two extracting procedures (Soxhlet and sonication) was
also compared in order to suggest the best analytical
procedure for the determination of the pollutants in the
soils.
2. Materials and methods
2.1. The contaminated site
The industrial area of Cengio (near Savona) in
Northern Italy is extremely polluted due to irregular
disposal of contaminants on surface and lower soil
horizons since 1882, when SIPE (Societa` Italiana
Prodotti Esplodenti) began production of explosives.
Later, SIPE became ACNA (Aziende Chimiche Nazionali Associate) that produced phenol, sulphuric acid,
nitric acid and several other industrial organic intermediates. In 1939, ACNA additionally manufactured
a range of colouring organic compounds (more information at: http://www.bonicare.it/). The serious
environmental contamination (soils and waters) around
the ACNA site determined in 1999 its inclusion in the
list of national priorities for environmental reclamation
and fostered a number of activities among which
a scientic research on soil remediation led by the
Interuniversity Consortium Chemistry for the Environment (INCA, Venice, Italy; http://helios.unive.it/inca).
2.2. Soils
Two dierent soil samples (A and B) were sampled in
the ACNA site. After air-drying, the soils were rst 2.00
mm sieved, and, then, analysed for their texture and
content of carbon and nitrogen (Fisons EA 1108
Elemental Analyzer). The values are reported in Table 1.

P. Conte et al. / Environmental Pollution 135 (2005) 515522

Table 1
Characteristics of soil A and B from the contaminated ACNA site at
Cengio

Coarse sand (%)

Fine sand (%)
Silt (%)
Clay (%)
Carbon (%)
Nitrogen (%)

Soil A

Soil B

21.2G0.6
35.6G0.8
39.6G0.5
3.6G0.2
2.8G0.1
1.2G0.1

12.8G0.4
38.8G0.5
43.6G0.7
4.8G0.3
2.7G0.2
0.13G0.02

2.3. Reagents for soil washings

Solutions used for soil washings were: 1. MilliQgrade water; 2. 4% (w/v) sodium dodecylsulphate
(SDS); 3. 4% (v/v) Triton X-100 (TX100) solutions; 4.
10 mg/l humic acid (HA). The latter is reported to be the
critical micelle concentration of HAs (Guetzlo and
Rice, 1994). SDS and Triton were obtained from
Sigma (Milano, Italy) and used without further
purication. HA was isolated from a North Dakota
Leonardite (Mammoth, Chem. Co., Houston, Texas),
and puried as reported elsewhere (Conte and Piccolo,
1999). The HA was then suspended in distilled water
and titrated to pH 7 by an automatic titrator (VIT 909
Videotitrator, Copenhagen) with a 0.1 M NaOH
solution under N2 stream. The resulting sodium-humate
was then ltered through a Millipore 0.45 m and freezedried.
2.4. Soil washing
Soils (10 g) were suspended in 100 mL of each soil
washing solution, and shaken for 24 h in a rotary shaker
at 10 rpm. All suspensions were centrifuged in glass
tubes at 10,000 rpm for 10 minutes in order to separate
solid residues from washing solutions. The solid residues
were dried at 40  C and subjected to either ultrasonication or Soxhlet extraction.
2.5. Ultrasonic extraction
Each soil sample (10 g) either before or after the
washing was suspended in 100 mL of an acetone/nhexane (1:1) mixture, and sonicated with a Misonix
XL2020 sonicator. A power of 55 Watt was applied for
12 minutes to the soil suspensions to obtain a total
energy of 39.6 kJ. After sonication, the suspension was
ltered through Whatman 40 paper lters, in order to
separate the residue from the extract.

517

mixture in a Soxhlet equipment for 48 hours (Dean,

1999). According to the EPA SW 3630 B method, the
acidic organic contaminants (such as organic acids and
phenols) were removed from the extract by a liquidliquid extraction with NaOH at pH 11. The organic
phase was treated with Na2SO4 for the removal of
residual water, and ltered through a Whatman 40 lter.
2.7. Sample pre-treatment
After ltration, the organic extracts coming from
either sonication or Soxhlet, were rst dried in a rotoevaporator at 40  C, and re-dissolved in 5 mL of
CH2Cl2. 1 mL of the CH2Cl2 solution was puried by
solid phase extraction (SPE) through elution in BondElut C-18 cartridges (500 mg/3 mL from Varian) with 30
mL of n-hexane, according to EPA SW 8270 B method.
The eluted n-hexane fraction was dried in a rotoevaporator at 40  C and re-dissolved in 1mL of CH2Cl2
and 1 mL of a 100 mg/mL octauoronaphtalene solution
in CH2Cl2, used as internal standard. The concentration
of contaminants identied by GC-MS was then relative
to that of octauoronaphtalene.
2.8. GC-MS analysis
A Perkin-Elmer Autosystem XL gas-chromatograph, equipped with a Programmed-Temperature
Split/Splitless injector with programmable pneumatic
control kept at a constant temperature of 250  C,
a Restek Rtx-5MS capillary column (5% diphenyl95% dimethylpolysiloxane, length 30 m, 0.25 mm ID,
and 0.25 mm df), and a Perkin-Elmer TurboMass Gold
mass-spectrometer, was used for qualitative and quantitative analysis of contaminants extracted from the
original soils and their residues after the washings. The
conditions used for GC analyses were the following: 1.
initial temperature of 40  C for 5 minutes; 2. to 250  C
at a 3  C/min rate; 3. isothermal for 20 minutes. The
total GC run time was 95 minutes. Helium was the
carrier gas at 1.5 mL/min with a split-ow of 30mL/min.
The inlet-line temperature of the GC-MS system was set
at 280  C, while that of the MS source at 180  C. A
solvent delay time of 5 minutes was applied before
acquisition of the mass spectra to prevent lament
injuries. Low and high m/z limits of the mass
spectrometer were 50, and 400 amu, respectively. A
NIST mass spectral library version 1.7 was used for
peak identication.
2.9. Data treatment

2.6. Soxhlet extraction

Each soil sample (10 g) either before or after the
washing was weighed in Soxhlet lters, and extracted
under reux with 100 mL of an acetone/n-hexane (1:1)

Soxhlet and sonication extractions were performed in

triplicate for each soil before and after soil washings.
Each organic extract was analysed in duplicate by
GC-MS analysis. Quantitative results by GC-MS

518

P. Conte et al. / Environmental Pollution 135 (2005) 515522

analyses were weight-averaged to provide experimental

error.

3. Results and discussion

3.1. Eect of soil texture on contaminants
extractability
Soxhlet and sonication are the most common liquidsolid extractive procedures used to obtain pollutants
from soil samples (Dean, 1999). The former is a heatrequiring method, whereas sonication is based on the
ultrasonic shaking of solid suspensions (Martens et al.,
2002). Both methods remove semivolatile, and nonvolatile organics from soils (US EPA methods 3540, and
3550). However, the removing eciency of both
methods may be strongly aected by soil characteristics
and especially by soil texture (Jastrow, 1996; Peerkorn,
1997). It is widely accepted that a ne-sized texture in
soil is a main factor for the hydrophobic adsorption of
organic matter on the surface of ne colloidal soil
particles, thereby favouring the associations of microinto macro-aggregates (Jastrow, 1996; Peerkorn, 1997;
Piccolo and Mbagwu, 1999). Concomitantly, the organic
material trapped into the macroaggregates is physically
protected from biodegradation and becomes hardly
extractable by common extraction procedures, unless
the soil macroaggregates are disrupted by applying either
mechanical or sonication energy (Peerkorn, 1997;
Watts et al., 2000; Mulder et al., 2001; Plante and
McGill, 2002). Conversely, coarse-sized soil particles are
less prone to form larger soil aggregates mediated by
organic matter (Kemper and Rosenau, 1986). Thus,
organic compounds are located only on the surface of the
coarse-sized fractions and result removable with the
same eciency by both soxhlet and sonication (Peerkorn, 1997; Watts et al., 2000; Mulder et al., 2001).
For the two ACNA soils, the texture of sample A
resulted larger in coarse-sized particles, whereas that of
soil B was higher in ne-sand, silt and clay (Table 1).
Concomitantly, the two soils resulted dierent in the
amount of organic contaminants that could be extracted

by either soxhlet or sonication (Table 2). Soil A was

richer in polycyclic aromatic hydrocarbons (PAH), and
monoaromatic halogenated and nitrogenated compounds and soil B had a larger content of thiophenes,
sulphones, and biphenyls (Table 2). However, the total
amount of respective contaminants extracted from soil
A by soxhlet and sonication was not signicantly
dierent, whereas that of soil B was more exhaustively
extracted by sonication (Table 2).
This dierent eciency of extractions methods
between soil A and B may be attributed to their dierent
texture. The lower aggregate stability conferred to soil A
by its larger content of coarse-sized fractions allowed
a similar availability to both soxhlet and sonication
extractions of organic compounds adsorbed on the
outer surfaces of gross particles, thereby resulting in
similar extracting eciencies (Table 2). Conversely, in
the ne-textured soil B, organic pollutants are adsorbed
on the surfaces of ne particles contained in more
structurally stable macroaggragates. Hence, the soxhlet
extraction, based simply on temperature, could only
wash away the fraction of contaminants mainly present
on the surfaces of macroaggregates, whereas ultrasonic
vibrations, by exerting a disaggregation energy on the
macroaggregates (Babic et al., 1998; Martens et al.,
2002), could also expose to solubilization the pollutants
adsorbed on the surfaces of the inner ne particles. For
this reason, the amount of pollutants extracted by
sonication from soil B resulted larger than that obtained
by soxhlet (Table 2). An exception were the thiophenes
that were obtained from soil B in equal amount both by
soxhlet and sonication. This suggests that thiophenes
were preferentially adsorbed mainly on the external
surface of the soil aggregates, thereby being equally
dissolved by solvent both in soxhlet and sonication.
3.2. Soil washings
The washing with just water was found not to
eciently remove pollutants from soil A (Fig. 1). In
fact, the totality of pollutants obtained from soil A, both
by soxhlet and sonication after washing with Milli-Q
grade water, was only slightly lower than that recovered

Table 2
Amount (mg kg1) of specic contaminants extracted from soil A and soil B by both soxhlet and sonication
Contaminants

Soxhlet

Sonication

Soxhlet

PAH
Monoaromatic halogenated
and nitrogenated compounds
Thiophenes
Sulphones
Biphenyls
Other

Soil A

Soil A

Soil B

4562G445
407G22

3792G130
426G19

37G4
211G27
108G16
78G6

28G2
227G10
95G7
96G6

986G129
97G3
2337G98
243G12
16G1
31G10

Sonication
Soil B
1337G152
222G14
2815G344
360G41
152G7
47G5

519

P. Conte et al. / Environmental Pollution 135 (2005) 515522

7000

Soil A

6000
5000
4000
3000

Concentration (mg.kg-1)

2000
1000
0
Soxhlet

6000

Sonication
5000

Soil B

4000
3000
2000
1000

SW-HA

SW-TX100

SW-SDS

SW-H2O

Soil

Fig. 1. Total amount of contaminants extracted by both soxhlet and

sonication from soil A and soil B and after soil washing (SW) with
H2O, SDS, TX100, and HA.

from the unwashed soil. Washing with water appeared

more ecient in removing pollutants from soil B,
especially when sonication was used for extraction
procedure. This may be attributed in this ne-textured
soil to a process of soil particle aggregation in water
(Oades and Waters, 1991; Christensen, 1992; Puget
et al., 2000) that limits pollutant solubilization by
soxhlet.
Synthetic surfactants are reported to remove organic
pollutants from contaminated soils by a mechanism

involving repartition of pollutants into the surfactant

micellar phases formed in water. However, this process
would take place only when the surfactant solution
reaches the surfaces of soil particles and the pollutants
may be desorbed into the micellar phases (Noordman
et al., 2000; Chun et al., 2002; Cuypers et al., 2002;
Mata-Sandoval et al., 2002).
The soil washings conducted in this work by either
synthetic (SDS and TX100) or natural surfactants (HA)
were more ecient than water in removing signicant
amounts of pollutants from the highly contaminated
soils (Fig. 1). In general, the synthetic surfactants used
here were able to reduce the content of contaminants
from the two soils from 80 to more than 90% and
conrmed previous literature ndings (Chefetz et al.,
2000; Holman et al., 2002; Kim and Lee, 2002). This
work also indicates for the rst time that a natural nontoxic surfactant such as humic acid removes similar
amounts of contaminants from a polluted soil as
synthetic surfactants (Fig. 1).
Soil washing (SW) may aect soil structural hierarchy
by redistributing the size of aggregate particles (Kemper
and Rosenau, 1986). No signicant dierence was
observed in the amount of PAHs, biphenyls, hydrophilic
halogenated, and nitrogenated compounds, thiophenes,
and sulphones obtained by soxhlet and sonication after
SW of soil A with H2O (Table 3). Hence, the predominantly coarse texture of soil A did not produce any
further particle aggregation in water that aects the
removal of contaminants. The ner texture of soil B,
instead, and the consequent tendency of soil particles to
create aggregation, may be the reason for the larger
amount of pollutants removed by sonication than by
soxhlet (Table 4).
The use of either synthetic or natural surfactants in
SW techniques may also have the eect of increasing soil
structural stability due to the aggregating action that the
hydrophobicity of surfactants may exert on the solid soil
particles (Piccolo and Mbagwu, 1999). This may in turn
aect the solubilization of contaminants during the
determination procedures depending on the specic
structure of pollutants.
A change in soil aggregation after soil washings may
explain the results on the removal of specic pollutants.

Table 3
Amount (mg kg1) of pollutants extracted from soil A by both soxhlet and sonication after soil washing (SW) with H2O, SDS, TX100, and HA
Pollutants

SW-H2O
soxhlet

Sonication

soxhlet

sonication

soxhlet

sonication

soxhlet

sonication

PAH
Monoaromatic halogenated
and nitrogenated compounds
Thiophenes
Sulphones
Biphenyls
Other

3071G185
167G10

3607G482
172G18

1033G16
95G4

1639G158
75G9

1366G92
93G7

1944G73
87G11

1411G83
60G4

1318G286
83G7

6G1
175G4
43G2
52G2

23G4
103G5
27G6
41G6

11G1
51G1
33G1
27G1

0
53G4
10G1
23G3

9G2
65G3
6G1
23G4

11G1
56G2
16G1
71G3

2G1
45G7
31G2
29G4

SW-SDS

SW-TX100

15G1
40G3
17G1
23G34

SW-HA

520

P. Conte et al. / Environmental Pollution 135 (2005) 515522

Table 4
Amount (mg kg1) of pollutants extracted from soil B by both soxhlet and sonication after soil washing (SW) with H2O, SDS, TX100, and HA
Pollutants

PAH
Monoaromatic halogenated
and nitrogenated compounds
Thiophenes
Sulphones
Biphenyls
Other

SW-H2O

SW-SDS

SW-TX100

SW-HA

soxhlet

sonication

soxhlet

sonication

soxhlet

sonication

soxhlet

sonication

419G5
67G3

1340G20
102G4

338G29
54G4

496G43
44G6

600G41
62G4

583G109
30G2

340G25
18G6

666G33
66G3

421G9
35G1
11G1
15G1

563G50
71G2
16G1
14G1

233G22
27G1
9G1
17G1

282G20
71G7
8G1
15G3

485G29
32G3
11G1
15G1

591G60
28G2
6G1
3G1

396G68
17G7
0
16G2

691G18
50G8
10G2
28G1

For soil A, after washing with SDS, the PAHs were

subsequently more eciently removed by sonication
than by soxhlet (Table 3). Conversely, soxhlet allowed
to obtain larger amounts of thiophenes, biphenyls, and
monoaromatic nitrogenated and halogenated compounds (Table 3). After soil washing with TX100,
sonication provided again larger amounts of hydrophobic PAHs and sulphones, whereas soxhlet appeared
more ecient in obtaining thiophenes, and biphenyls
(Table 3). These ndings may be attributed to a variation
in the aggregation of soil particles after treatment with
surfactants and a consequent dierent distribution of
pollutants in either inner or outer surfaces of new soil
aggregates.
Also in the case of SW with HA, the treatment with
this natural organic matter may have produced an
alteration in aggregate distribution of soil A and
consequent solubilization of dierent contaminants.
However, in contrast to SDS and TX100, washing of
soil A with HA was able to remove a signicantly
similar and large amount of PAHs by either soxhlet or
sonication (Table 3). This may be attributed to the
heterogeneity of the HA micellar phase and, hence, its
capacity to incorporate with equal eciency PAHs of
dierent structures (Fava and Piccolo, 2002). Nevertheless, sonication removed more monoaromatic nitrogenated and halogenated compounds, and biphenyls
than soxhlet, whereas the latter technique was more
ecient in obtaining thiophenes (Table 3).
More than in soil A, alteration of soil aggregation in
the ner-textured soil B due to washings appears to be the
explanation of the removal of specic pollutants. Except
for the SW in TX100, all other washings determined
a subsequent removal of larger quantities of PAHs by
sonication (Table 4). Sulphones, the larger contaminant
in soil B (Table 2), were signicantly removed in larger
amounts by sonication than by soxhlet after SW with all
solutions (Table 4). With few exceptions, the rest of
specic contaminants found in soil B were more
quantitatively removed by sonication after all SW
procedures. This general more ecient extracting action
of sonication for soil B after the SW in the dierent
solution is parallel to that shown for the unwashed soil.
Again, this is to be attributed to a larger structural

stability of this soil whose inteparticle association energy

is overcome by application of sonication, thereby
allowing a solubilization of specic pollutants impossible
to reach by a simple soxhlet extraction.

4. Conclusions
This work has conrmed that water cannot be
recommended for an ecient removal of pollutants
from a contaminated soil and that organic surfactants
should be relied upon in soil washing procedures.
However, synthetic surfactants such as SDS, and
TX100 which are ecient in soil washings may become
a further environmental problem because of their
biological toxicity. We have shown here that a natural
surfactant such as a humic acid solution can be used in
washings of a contaminated soil with the same eciency
as that of synthetic surfactants. The importance to
promote the use of environmentally-safe humic substances in the remediation of heavily contaminated soils
should be self-evident. Humic substances can even
improve biomass activity on washed soils and contribute
to a further natural attenuation once the soil are
disposed after an ex situ remediation process (Fava
and Piccolo, 2002; Marschner and Kalbitz, 2003).
Moreover, the fraction of humic substances remaining
in soil may play a favourable action in plant growth and
thus assist in the full recuperation of the treated soils
(Nardi et al., 2002).
Our ndings have also indicated that soil texture is an
important parameter to take into account to select the
most appropriate analytical technique to remove and
determine soil contaminants. We found that contaminants are more quantitatively removed by sonication
rather than by soxhlet when a soil is characterized by
a ne texture, whereas both soxhlet and sonication give
equivalent results in more coarse-textured soils. However, since sonication is faster than soxhlet (in our
experimental work it took only 12 minutes against 48
hours needed for soxhlet) the use of sonication should
be recommended as a standard procedure for all kind
of soils. The disadvantage of sonication, a reported
degradation of aromatic organic pollutants (Goskonda

P. Conte et al. / Environmental Pollution 135 (2005) 515522

et al., 2002), is reasonably balanced by its more

quantitative removal eciency.
Acknowledgements
The authors acknowledge Consorzio Interuniversitario La Chimica per lAmbiente (INCA) and Ministero dellAmbiente that funded this work through the
Progetto Sisifo.

References
Babic, S., Petrovic, M., Kastelan-Macan, M., 1998. Ultrasonic solvent
extraction of pesticides from soil. J. Chromat. A 823, 39.
Ballarin-Denti, A., Bertazzi, P.A., Facchetti, S., Fanelli, R., Mocarelli,
P., 1999. Chemistry, Man and Environment. The Severo Accident
20 Years on: Monitoring, Epidemiology and Remediation.
Elsevier, Oxford, UK.
Boopathy, R., 2000. Factors limiting bioremediation technologies.
Biores. Technol. 74, 6367.
Borga, K., Gabrielsen, G.W., Skaare, J.U., 2001. Biomagnication of
organochlorines along a Barents Sea food chain. Environ. Poll.
113, 187198.
Chefetz, B., Deshmukh, A.P., Hatcher, P.G., 2000. Pyrene sorption by
natural organic matter. Environ. Sci. Technol. 34, 29252930.
Chin, Y.-P., Aiken, G.R., Danielsen, K.M., 1997. Binding of pyrene to
aquatic and commercial humic substances: the role of molecular
weight and aromaticity. Environ. Sci. Technol. 31, 16301635.
Christensen, B.T., 1992. Physical fractionation of soil and organic
matter in primary particle size and density separates. Adv. Soil Sci.
20, 190.
Chu, W., Chan, K.H., 2003. The mechanism of the surfactant-aided
soil washing system for hydrophobic and partial hydrophobic
organics. Sci. Tot. Environ. 307, 8392.
Chun, C.L., Lee, J.-J., Park, J.-W., 2002. Solubilization of PAH
mixtures by three dierent anionic surfactants. Environ. Poll. 118,
307313.
Conte, P., Piccolo, A., 1999. Conformational arrangement of dissolved
humic substances. Inuence of solution composition on association
of humic molecules. Environ. Sci. Technol. 33, 16821690.
Conte, P., Zena, A., Pilidis, G., Piccolo, A., 2001. Increased retention
of polycyclic aromatic hydrocarbons in soils induced by soil
treatment with humic substances. Environ. Poll. 112, 2731.
Cuypers, C., Pancras, T., Grotenhuis, T., Rulkens, W., 2002. The
estimation of PAH bioavailability in contaminated sediments using
hydroxypropyl-b-cyclodextrin and Triton X-100 extraction techniques. Chemosphere 46, 12351245.
Dean, J.R., 1999. Extraction Methods for Environmental Analysis.
Wiley and Sons, New York.
Deshpande, S., Shiau, B.J., Wade, D., Sabatini, D.A., Harwell, J.H.,
1999. Surfactant selection for enhancing ex situ soil washing. Wat.
Res. 33, 351360.
Fava, F., Piccolo, A., 2002. Eect of humic substances on the
bioavailability aerobic biodegradation of polychlorinated biphenyls in a model soil. Biothecnol. Bioeng. 77, 204211.
Fent, K., 2003. Ecotoxicological problems associated with contaminated sites. Toxicol. Lett. 140/141, 353365.
Gaboriau, H., Saada, A., 2001. Inuence of heavy organic pollutants
of anthropic origin on PAH retention by kaolinite. Chemosphere
44, 16331639.
Garon, D., Krivobok, S., Wouessidjewe, D., Seigle-Murandi, F., 2002.
Inuence of surfactants on solubilization and fungal degradation of
uorine. Chemosphere 47, 303309.

521

Goskonda, S., Catallo, W.J., Junk, T., 2002. Sonochemical degradation of aromatic organic pollutants. Waste Manag. 22, 351356.
Griths, R.A., 1995. Soil washing technology and practice. J. Haz.
Mat. 40, 175189.
Guetzlo, T.F., Rice, J.A., 1994. Does humic acid form a micelle? Sci.
Total Environ. 152, 3135.
Halim, M., Conte, P., Piccolo, A., 2003. Potential availability of heavy
metals to phytoextraction from contaminated soils induced by
exogenous humic substances. Chemosphere 52, 265275.
Holman, H.-Y.N., Nieman, K., Sorensen, D.L., Miller, C.D., Martin,
M.C., Borch, T., McKinney, W.R., Sims, R.C., 2002. Catalysis of
PAH biodegradation by humic acid shown in synchroton infrared
studies. Environ. Sci. Technol. 36, 12761280.
Huang, W., Peng, P., Yu, Z., Fu, J., 2003. Eects of organic matter
heterogeneity on sorption and desorption of organic contaminants
by soils and sediments. Applied Geochem. 18, 955972.
Janzen, R.A., Xing, B., Gomez, C.C., Salloum, M.J., Drijber, R.A.,
McGill, W.B., 1996. Compost extract enhances desorption of
naphthol and naphthalene from pristine and contaminated soils.
Soil Biol. Biochem. 28, 10891098.
Jastrow, J.D., 1996. Soil aggregate formation and the accrual of
particulate and mineral-associated organic matter. Soil Biol.
Biochem. 28, 665676.
Kemper, W.D., Rosenau, R.C., 1986. Aggregate stability and size
distribution. In: Klute, A. (Ed.), Methods of Soil Analysis, Part I,
second ed. Agron. Monogr. 9. ASA ans SSSA, Madison, WI, pp.
425442.
Kiem, R., Knicker, H., Ligouis, B., Kogel-Knabner, I., 2003. Airborne
contaminants in the refractory organic carbon fraction of arable
soils in highly industrialized areas. Geoderma 114, 109137.
Kim, Y., Lee, D., 2002. Solubility enhancement of PCDD/F in the
presence of dissolved humic matter. J. Haz. Mat. B91, 113127.
Kopinke, F.-D., Georgi, A., Mackenzie, K., 2000. Sorption and
chemical reactions of PAHs with dissolved humic substances and
related model polymers. Acta Hydrochim. Hydrobiol. 28, 385399.
Kuhlman, M.I., Greeneld, T.M., 1999. Simplied soil washing
processes for a variety of soils. J. Haz. Mat. 66, 3145.
Lee, D.-H., Cody, R.D., Kim, D.-J., Choi, S., 2002. Eect of soil
texture on surfactant-based Remediation of hydrophobic organiccontaminated soil. Environ. Int. 27, 681688.
Mackay, D., Fraser, A., 2000. Bioaccumulation of persistent organic
chemicals: mechanisms and models. Environ. Poll. 110, 375391.
Mann, M.J., 1999. Full-scale and pilot-scale soil washing. J. Haz. Mat.
66, 119136.
Marschner, B., Kalbitz, K., 2003. Controls of bioavailability and
biodegradability of dissolved organic matter in soils. Geoderma
113, 211235.
Martens, D., Gfrerer, M., Wenzl, T., Zhang, A., Gawlik, B.M.,
Schramm, K.W., Lankmayr, E., Kettrup, A., 2002. Comparison of
dierent extraction techniques for the determination of polychlorinated organic compounds in sediments. Anal. Bioanal. Chem. 372,
562568.
Mata-Sandoval, J.C., Karns, J., Torrents, A., 2002. Inuence of
Rhamnolipids and Triton X-100 on the desorption of pesticides
from soils. Environ. Sci. Technol. 36, 46694675.
Mulder, H., Breure, A.M., Rulkens, W.H., 2001. Application of
a mechanistic desorption-biodegradation model to describe the
behavior of polycyclic aromatic hydrocarbons in peat soil
aggregates. Chemosphere 42, 285299.
Nardi, S., Sessi, E., Pizzeghello, D., Sturaro, A., Rella, R., Pargoli, G.,
2002. Biological activity of soil organic matter mobilized by root
exudates. Chemosphere 46, 10751081.
Noordman, W.H., Bruining, J.-W., Wietzes, P., Janssen, D.B., 2000.
Facilitated transport of a PAH mixture by a rhamnolipid biosurfactant in porous silica matrices. J. Contam. Hydr. 44, 119140.
Oades, J.M., Waters, A.G., 1991. Aggregate hierarchy in soils. Austr.
J. Soil Sci. 29, 815828.

522

P. Conte et al. / Environmental Pollution 135 (2005) 515522

Ockenden, W.A., Breivik, K., Meijer, S.N., Steinnes, E., Sweetman,

A.J., Jones, K.C., 2003. The global re-cycling of persistent organic
pollutants is strongly retarded by soils. Environ. Poll. 121, 7580.
Paaso, N., Peuravuori, J., Lehtonen, T., Pilhaja, K., 2002. Sedimentdissolved organic matter equilibrium partitioning of pentachlorophenol: the role of humic matter. Environ. Int. 28, 173183.
Peerkorn, E., 1997. Structure and stability of natural organic matter/
soil complexes and related synthetic and mixed analogues. Adv.
Coll. Int. Sci. 73, 127200.
Peuravuori, J., 2001. Binding of pyrene on lake aquatic humic matter:
the role of structural descriptors. Anal. Chim. Acta 429, 7589.
Peuravuori, J., Paaso, N., Pilhaja, K., 2002. Sorption behaviour of some
chlorophenols in lake aquatic humic matter. Talenta 56, 523538.
Piccolo, A., Mbagwu, J.S.C., 1999. Role of hydrophobic components
of soil organic matter in soil aggregate stability. Soil Sci. Soc. Am.
J. 63, 18011810.
Piccolo, A., Celano, G., Conte, P., 1996. Adsorption of glyphosate by
humic substances. J. Agric. Food Chem. 44, 24422446.
Plante, A.F., McGill, W.B., 2002. Soil aggregate dynamics and the
retention of organic matter in laboratory-incubated soil with
diering simulated tillage frequencies. Soil Till. Res. 66, 7992.
Puget, P., Chenu, C., Balesdent, J., 2000. Dynamics of soil organic
matter associated with particle-size fractions of water-stable
aggregates. Eur. J. Soil Sci. 51, 595605.

Sandbacka, M., Christianson, I., Isomaa, B., 2000. The acute toxicity
of surfactants on sh cells, Daphnia magna and sh a comparative
study. Tox. Vitro 14, 6168.
Schwarzenbach, A.R., Gschwend, P.M., Imboden, D.M., 2003.
Environmental Organic Chemistry. second ed. J. Wiley and Sons,
New York.
Simcik, M.F., Eisenreich, S.J., Lioy, P.J., 1999. Source
apportionment and source/sink relationships of PAHs in the
coastal atmosphere of Chicago and Lake Michigan. Atm. Environ.
33, 50715079.
Venkataraman, C., Negi, G., Sardar, S.B., Rastogi, R., 2002.
Size distributions of polycyclic aromatic hydrocarbons in
aerosol emissions from biofuel combustion. Aer. Sci. 33,
503518.
Watts, C.W., Eich, S., Dexter, A.R., 2000. Eects of mechanical
energy inputs on soil respiration at the aggregate and eld scales.
Soil Till. Res. 53, 231243.
Weng, L., Temmingho, E.J.M., Lofts, S., Tipping, E., Van
Riemsdijk, W.H., 2002. Complexation with dissolved organic
matter and solubility control of heavy metals in a sandy soil.
Environ. Sci. Technol. 36, 48044810.
Zhou, J.L., Rowland, S.J., 1997. Evaluation of the interactions
between hydrophobic organic pollutants and suspended particles
in estuarine waters. Wat. Res. 31, 17081718.