Environmental Pollution 222 (2017) 348e355

Contents lists available at ScienceDirect

Environmental Pollution
journal homepage: www.elsevier.com/locate/envpol

Immobilization of metals in contaminated soils using natural

polymer-based stabilizers*
Xue Tao, Aimin Li, Hu Yang*
State Key Laboratory of Pollution Control and Resource Reuse, School of the Environment, Nanjing University, Nanjing 210023, PR China

a r t i c l e i n f o a b s t r a c t

Article history: Three low-cost natural polymer materials, namely, lignin (Ln), carboxymethyl cellulose, and sodium
Received 17 September 2016 alginate, were used for soil amendment to immobilize lead and cadmium in two contaminated soil
Received in revised form samples collected from a mining area in Nanjing, China. The remediation effects of the aforementioned
3 November 2016
natural polymers were evaluated by toxicity characteristic leaching procedure (TCLP) and sequential
Accepted 6 December 2016
extractions. The stabilizers could lower the bioavailability of Pb and Cd in the contaminated soils, and the
Available online 24 December 2016
amount of the exchangeable forms of the aforementioned two metals were reduced evidently. TCLP
results showed that the leaching concentrations of Pb and Cd were decreased by 5.46%e71.1% and 4.25%
Keywords:
Soil remediation
e49.6%, respectively, in the treated soils. The contents of the organic forms of the two metals both
Soil stabilizer increased with the increase in stabilizer dose on the basis of the redistribution of metal forms by
Lignin sequential extractions. These findings were due to the fact that the abundant oxygen-containing groups
Carboxymethyl cellulose on the polymeric amendments were effective in chelating and immobilizing Pb and Cd, which have been
Sodium alginate further confirmed from the metal adsorptions in aqueous solutions. Moreover, Ln achieved the greatest
Immobilization mechanism effect among the three polymers under study because of the former's distinct three-dimensional mo-
lecular structure, showing the preferential immobilization of Pb over Cd in soils also. Thus, the above-
mentioned natural polymers hold great application potentials for reducing metal ion entry into the
food chain at a field scale.
© 2016 Elsevier Ltd. All rights reserved.

1. Introduction
The soil serves as a significant part of the eco-environment and a
main resource for human survival. However, an increasing amount
of agricultural and urban soils have been polluted with the rapid
economic development and industrialization (Cheng, 2003; Shao
et al., 2014). Among various kinds of pollutants, metal pollution
have received great attention; in these phenomena, metals are
released to soils through various kinds of human activities, such as
mining, ore refinement, fertilizer industries, tannery, and pesticide
use (Bolan et al., 2014; Shao et al., 2014). Unlike organic pollutants,
metals are highly dangerous because of their toxicity and non-
biodegradability by soil organisms (Cheng, 2003; Wei and Yang,
2010). Such metals could exist in the soil for numerous years,
become accumulated through the food chain, and consequently
pose considerable threat to human health (Li et al., 2014). Among
*
This paper has been recommended for acceptance by Dr. Chen Da.
* Corresponding author.
E-mail address: yanghu@nju.edu.cn (H. Yang).
these metal contaminates, lead and cadmium are two highly pop-
ular and toxic examples (Jarup, 2003).
However, metal toxicity in soil depends not only on the total
amount but also on the mobility in soil. For a metal to be
bioavailable, it must first exist in a form that is mobile and acces-
sible to plants (Bolan et al., 2014). Bioavailability is greatly attached
to metal fractionation in soil, which frequently refers to the dis-
tribution of various metal forms, including exchangeable carbonate,
FeeMn oxide, organic, and residual forms (D'Amore et al., 2005;
Gleyzes et al., 2002; Tessier et al., 1979). Accordingly, the bioavail-
ability of metal contaminates in each form gradually decreases
from the exchangeable form to the residual form. Among the
different forms, the exchangeable form is the most dangerous.
Greater metal mobility is associated with higher adverse risk. Many
techniques have been developed and applied to treat metal-
polluted soils, such as the physical isolation method, biological
techniques, and stabilization/solidification method (Dermont et al.,
2008a, 2008b; Lageman et al., 2005; Liao et al., 2016; Ma et al.,
2001; Maini et al., 2000; Rahimi et al., 2013). Among such ap-
proaches, in situ solidification/stabilization has attracted great

http://dx.doi.org/10.1016/j.envpol.2016.12.028
0269-7491/© 2016 Elsevier Ltd. All rights reserved.
 X. Tao et al. / Environmental Pollution 222 (2017) 348e355
attention because of its high efficiency and cost-effectiveness
(Bolan et al., 2014; Guo et al., 2006; Kumpiene et al., 2008; Sun
et al., 2012; Tica et al., 2011; USEPA, 2007.). The final stabilization
performance of the targeted contaminated soils largely depends on
the selected immobilization agents. Moreover, many external pa-
rameters, such as pH, age of contamination, soil types, cation ex-
change capacity (CEC), coexisting natural organic matters, and
other inorganic contaminants in soils, also affect the remediation
efficiency (Bolan et al., 2014; Harter, 1983; Lee et al., 2009). The use
of many kinds of soil stabilizers, including lime, phosphate mate-
rials, zeolite, bentonite, sepiolite, Fe/Mn oxides, and organic mat-
ters, have been attempted (Gonza
lez-Nún
~ ez et al., 2011; Guo et al.,
2006; Kumpiene et al., 2008; Lee et al., 2009; Mallampati et al.,
2012; Mignardi et al., 2012; Querol et al., 2006; Sun et al., 2012).
Organic materials, such as bio-solid composts (VacaePaulin
et al., 2006) and biochar (Beesley et al., 2011), have constantly
attracted researcher attention as potential high-performance
immobilization agents for soil remediation (Bolan et al., 2014;
Calace et al., 2005; Dahrazma and Mulligan, 2007; Kumpiene
et al., 2008; Park et al., 2011). Although these organic materials
would decompose after a long period of stabilization and holds
certain remobilization risks, they could decrease toxic metal
bioavailability and induce metal retention in soils by adsorption
and facile formation of stable complexes with metals (Guo et al.,
2006). Such effect could also improve soil quality by providing
nutrients to plants. Among numerous organic materials, lignin (Ln)
is the main byproduct of the paper industry and the precursor of
humic substances; Ln contains abundant oxygen-containing
groups, including carboxyl, lactonic, and phenolic hydroxyl
groups, thereby easily chelating and immobilizing many metals
(Jiang, 2009; Zhou and Haynes, 2010). Given its unique chemical
composition and richness in nature, Ln holds a significant potential
as an effective soil stabilizer (Jiang, 2009; Zhou and Haynes, 2010).
However, data related to the stabilizing behaviors of Ln in reme-
diating toxic-metal-polluted soils have not been systematically
reported to date, especially its remediation mechanisms (Battaglia
et al., 2007; Calace et al., 2005).
In the current study, Ln was employed to treat actual Pb- and Cd-
polluted soils collected near the mining areas of Nanjing, China. For
comparison, two other organic materials, namely, carboxymethyl
cellulose (CMC) and sodium alginate (SA), were adopted. Both
materials are polysaccharides containing abundant carboxyl
groups. We investigated and compared in detail the remediation
performance of the aforementioned three natural polymer stabi-
lizers for metal-contaminated soils. Furthermore, we studied and
discussed herein the relevant mechanisms of Ln, CMC, and SA in the
immobilization processes by combination of metal adsorption in
aqueous solutions, including their competitive immobilizations.
2. Materials and methods
2.1. Materials
2.1.1. Contaminated soils
Two polluted soil samples were collected from the vicinity of
Qixiashang lead-zinc mines in Nanjing, China, which is a poly-
metallic deposit mainly containing lead and zinc associated with
sulfur and silver. As for the surrounding rocks of ores, NW plate
includes Huanglong formation at the bottom (C12) (coarse grained
and dolomitic limestones), Gaolishan group (C21) (siltstone), and
Hezhou group (C31) (marlite and calcareous shale); SE plate is
weakly mineralized limestone of Huanglong formation (C22). After
more than 50 years of mining activities, soils there have been
heavily polluted by various kinds of metals. Among them, Pb
(80e170 mg/kg) and Cd (7e18 mg/kg) were two mainly ones.
349
Surface soils were obtained at 0e20 cm depth and sieved to <2 mm
to remove impurities. Afterward, the samples were fully air dried
and finally stored in plastic bags prior to analysis. Meanwhile, the
Fourier transform infrared (FTIR) spectra of two different soil
samples were obtained using a FTIR spectrometer (Type TENSOR
27; Bruker Co.; Germany; Supplementary Materials Fig. S1(a)).
Accordingly, the two soil samples showed fairly similar character-
istics. Therefore, a series of techniques were applied to further
determine the physicochemical properties of the two soils
(Table 1). pH was measured at a soil:water ratio of approximately
1:2.5 (w/w) using a pH meter (Delta320, MettlereToledo. Switer-
land) (Bao, 1981). Organic matter content was determined through
the potassium dichromate volumetric method, whereas CEC was
analyzed by the ammonium acetate exchange method (Bao, 1981).
The soils were digested with a solution of HCleHNO3eHClO4 and
metal concentrations were determined using a thermo M6 atomic
absorption spectrometer.
2.1.2. Natural polymer amendments
Three natural polymers, Ln, CMC, and SA, were selected for soil
amendment, all of which were purchased from Nanjing Chemical
Reagent Co. Ltd. Ln was pretreated by NaOH to remove impurities
(Jiang, 2009). Epoxy chloropropane was employed as crosslinker to
increase the stability of CMC (Perkerson et al., 1960). Meanwhile, a
certain amount of calcium chloride was used to modify SA into a
more stable form (Kuo and Ma, 2001). After slight modifications,
the three organic materials were processed into more efficient and
stable immobilization agents.
FTIR spectra of Ln, CMC, and SA were obtained (Supplementary
Materials Fig. S1(b)) and suggested the possible existence of surface
functional groups in their molecular structures. Ln was composed
of various oxygen-containing functional groups. CMC and SA both
contain abundant carboxymethyl groups. In accordance with the
modified Boehm titration method (Boehm, 1994), the detailed
amounts of carboxyl, lactonic, and phenolic hydroxyl groups in Ln
were determined to be approximately 0.524, 0.053, and
0.628 mmol$g
1, respectively. For the other two natural polymers,
the total carboxyl groups of CMC and SA were 1.125 and
1.076 mmol$g
1, respectively.
2.1.3. Others
HCl, HNO3, HClO4, NaOH, CaCl2, and other chemical reagents
used in this work were all obtained from Nanjing Chemical Reagent
Co. Ltd. All the chemicals were of analytical grade, and distilled
water was used in all experiments.
2.2. Soil amendment experiments
2.2.1. Incubation experiments
100 g of air-dried soil samples were mixed with different sta-
bilizers thoroughly in polythene cups. The contents of the indi-
vidual amendments in each soil were maintained at 1, 2, 5, and
10 wt% to determine the optimal dose. A control sample without
stabilizer was also included. Four replicates were conducted for
each treatment, and each treated soil was incubated with 70%
water holding capacity at 25  C for 30 days. All the soils were
Table 1
The physicochemical properties of two soil samples from the vicinity of abandoned
mining areas.
Soil pH CEC Organic matter Total Pb Total Cd
(mmol$kg
1) (mg$kg
1) (mg$kg
1) (mg$kg
1)
Soil1 7.802 21.14 21.57 171.0 17.89
Soil2 7.478 18.27 15.81 80.12 7.490
350 X. Tao et al. / Environmental Pollution 222 (2017) 348e355
weighed every 3 days and watered to maintain moisture at a
constant level during the incubation period.
2.2.2. Toxicity characteristic leaching procedure (TCLP) extraction
After incubation, all the treated and untreated soils were
analyzed by TCLP extraction (Li et al., 2001). 1.0 g of soil samples
were extracted with acetic acid solution (adjusted to pH 2.88) for
18 h at 30 r$min
1. The supernatant was then collected for analysis
of metal concentrations by atomic absorption spectrometry.
2.2.3. Sequential extraction
The redistribution of metal forms was investigated by sequential
extraction (Tessier et al., 1979). Each of the chemical fractions was
operationally defined as follows: (1) exchangeable (E): dried soil
(1.0 g) extracted with 8.0 mL of 1.0 mol dm
3 MgCl2 (pH 7) in a
centrifuge tube for 1 h at room temperature with continuous
agitation; (2) carbonate (C): residue from the exchangeable fraction
extracted with 8.0 mL of 1.0 mol dm
3 NaOAc (pH 5.0 adjusted by
acetic acid) for 5 h at room temperature with continuous agitation;
(3) FeeMn oxide (FM): residue from the carbonate fraction
extracted with 20 mL of 0.04 mol dm
3 NH2OH$HCl in 25% acetic
acid (v/v) for 6 h at 96 ± 3  C with occasional agitation; (4) organic
(O): residue from the FeeMn oxide fraction extracted with 3.0 mL
of 0.02 mol dm
3 HNO3 and 5.0 mL of 30% H2O2 (pH 2) for 2 h at
85  C with occasional agitation and a subsequent second 3.0 mL of
30% H2O2 (pH 2) for 3 h at 85  C with occasional agitation. Then,
after cooling, 5.0 mL of 3.2 mol dm
3 NH4OAC in 20% HNO3 (v/v)
was added and the sample was diluted to 20 mL and agitated
continuously for 0.5 h; and (5) residual (R): residue from the
organic fraction was transferred to beakers and digested with
HCleHNO3eHClO4.
2.3. Batch adsorption experiments in aqueous solutions
To gain a better understanding of the remediation mechanism of
Ln, CMC and SA, their adsorption performance for removal of Pb
and Cd from aqueous solutions was carried out for comparison. The
fundamental adsorption behaviors of three natural polymers,
including the adsorption equilibrium and kinetics, are investigated
in detail at pH 5.0, since precipitations of metal hydroxides would
be appeared at pH higher than 6.0 which lead to inaccurate inter-
pretation of adsorption. The detailed experimental conditions were
described in Supplementary Materials Text S1.
3. Results and discussion
Two polluted soil samples were collected from the vicinity of the
abandoned mining areas in Nanjing, China. The total concentra-
tions of Pb and Cd were 80.12e171.0 and 7.490e17.89 mg/kg,
respectively (Table 1), which were both much higher than the
agricultural standard limitations according to the Environmental
Quality Standard for Soils of China (GB15618, 2008). This aspect
poses some challenges to the utility of such soils. The reduction in
metal solubility by chemical immobilization treatment can be
considered as a useful tool for reducing phytotoxicity and
improving the quality of contaminated soils. Then three natural
polymer stabilizers, Ln, CMC, and SA, have been selected and
employed to remediate such polluted soils.
The addition of organic stabilizers exerted some influences on
the physico-chemical properties of soils, such as pH. The stability of
metals in soil is usually pH dependent. Fig. 1 shows the pH of two
soils after usage of Ln, CMC, and SA. We found that the pH of the
two treated soil samples all increased slightly by about
0.078e0.535, which is favorable for the stabilization of metals due
to easy formation of metal hydroxides and enhanced chelating
effects between organic matter and metals. The mobility and
availability of metals decrease with increasing soil pH levels
(Kumpiene et al., 2008).
3.1. TCLP extraction
The results of TCLP extraction are listed in Table 2. Accordingly,
the Pb and Cd concentrations in TCLP leachate after addition of
various natural polymers were mostly decreased to some extent
compared with those of untreated soils. With the increase in sta-
bilizer dose, the TCLP concentrations of two metals both decreased
further. This result indicates that the three stabilizers could achieve
good immobilization effects on Pb and Cd in soils. This effect was
attained because the abundant oxygen-containing groups on the
natural polymers exhibit good affinity and efficient chelating ef-
fects for the metals (Conte et al., 2005; Karlsson et al., 2007). The
good metal immobilization effects of the natural organic matters
have been also confirmed by Battaglia et al. (2007). They studied
the metal stabilization behaviors of a paper mill sludge, in which
organic matter consisted mainly of lignin, tannins and cellulose.
The addition of paper mill sludge to soil contaminated by Pb and Zn
induces a decrease in the mobile forms of both metals.
Moreover, the adsorption equilibrium and kinetics of Pb and Cd
on three natural polymers in aqueous solutions have been also
conducted at 298 K and pH 5.0 for a better understanding of the
immobilization mechanism (Supplementary Materials Fig. S2). The
aforementioned results have been further analyzed by various
isothermal and kinetic models (Freundlich, 1906; Ho and McKay,
1998; Lagergren, 1898; Langmuir, 1918), which results were listed
at Supplementary Materials Table S1 and S2, respectively.
Comprehensively considering the correlation coefficients of those
fitting models showed in Table S1 and S2 as well as the molecular
features of the adsorbents and adsorbates (Jiang, 2009; Zhou and
Haynes, 2010), the adsorption isotherms all closely followed
Langmuir model and the adsorption kinetics were well described
by the pseudo-second order equation. In general, this indicates that
the adsorption behaviors were a monolayer chemical adsorption. It
further confirmed that good immobilization effects on Pb and Cd in
soils was due to the efficient chelating effects between various
oxygen-containing groups on those natural polymers and metals
(Conte et al., 2005; Karlsson et al., 2007).
For further comparison, the Kds of Pb and Cd, which describe the
immobilization efficiency of the stabilizers, were calculated on the
basis of Eq. (1) and listed in Table 3.
m0
mf
Kd ¼ (1)
m0
where m0 and mf are the extracted concentrations of certain metals
in soil before and after stabilizer addition (Table 2). A higher Kd
signifies an efficient stabilizer.
In Tables 2 and 3, the immobilization effects of Ln, CMC, and SA
for the two metals were different from each other. For the reduction
of Pb in two soils, Ln showed much better effects than the other two
stabilizers. Meanwhile, the immobilization efficiency of Ln for Cd
was slightly lower than those of CMC and SA. Furthermore,
considering the comprehensive immobilization effects for both
metals from the sum of two Kds, i.e., Kd (Pb)þ Kd (Cd), Ln exerted
better immobilization effects on Pb and Cd and considerably
decreased the potential environmental risks. The differences in
immobilization effects among the three employed natural poly-
mers may be ascribed to their dissimilar oxygen-containing func-
tional groups and different chain architectures (inset of Fig. 1). The
three-dimensional structure of Ln may explain its higher efficiency
in sweeping and seizing metals from the soils than those of CMC
 X. Tao et al. / Environmental Pollution 222 (2017) 348e355 351

Fig. 1. Soil pHs before and after treated by various contents of different stabilization agents, Ln, CMC, and SA, respectively. The insets are the molecular structures of aforementioned
three natural polymers.

Table 2
The concentrations of targeted metals by TCLP extraction and their exchangeable form by sequential extraction in two different soil samples before and after amendment with
various contents of different stabilization agents.

By TCLP extraction

Pb (mg·kg¡1) Cd (mg·kg¡1)

0% 1% 2% 5% 10% 0% 1% 2% 5% 10%

Soil1 Ln 24.26 ± 0.01 17.32 ± 0.01 14.84 ± 0.08 14.51 ± 0.03 14.01 ± 0.02 1.32 ± 0.01 1.26 ± 0.01 1.05 ± 0.00 0.99 ± 0.00 0.95 ± 0.02
CMC 21.67 ± 0.00 20.95 ± 0.01 20.31 ± 0.01 19.21 ± 0.02 1.20 ± 0.01 0.86 ± 0.05 0.71 ± 0.01 0.71 ± 0.01
SA 22.30 ± 0.00 21.81 ± 0.01 20.80 ± 0.01 19.47 ± 0.01 1.29 ± 0.01 0.96 ± 0.02 0.86 ± 0.03 0.81 ± 0.02
Soil2 Ln 12.81 ± 0.01 7.43 ± 0.07 3.61 ± 0.05 NDa NDa 0.86 ± 0.02 0.81 ± 0.03 0.78 ± 0.02 0.76 ± 0.07 0.73 ± 0.03
CMC 12.49 ± 0.01 12.04 ± 0.00 10.85 ± 0.00 10.62 ± 0.01 0.82 ± 0.04 0.76 ± 0.02 0.75 ± 0.07 0.70 ± 0.05
SA 12.35 ± 0.00 12.11 ± 0.01 11.14 ± 0.00 11.06 ± 0.01 0.82 ± 0.09 0.78 ± 0.06 0.73 ± 0.05 0.71 ± 0.03

By sequential extraction

Pb (mg·kg¡1) Cd (mg·kg¡1)

0% 1% 2% 5% 10% 0% 1% 2% 5% 10%

Soil1 Ln 15.55 ± 0.10 12.83 ± 0.00 7.03 ± 0.07 4.10 ± 0.03 3.59 ± 0.03 1.20 ± 0.11 1.13 ± 0.10 0.94 ± 0.08 0.86 ± 0.01 0.81 ± 0.07
CMC 13.00 ± 0.00 12.32 ± 0.01 10.02 ± 0.01 9.56 ± 0.08 1.01 ± 0.07 0.91 ± 0.01 0.80 ± 0.02 0.71 ± 0.10
SA 14.56 ± 0.01 12.76 ± 0.01 11.74 ± 0.01 10.06 ± 0.05 1.04 ± 0.005 0.95 ± 0.02 0.87 ± 0.10 0.75 ± 0.02
Soil2 Ln 8.19 ± 0.07 7.76 ± 0.04 6.16 ± 0.02 6.10 ± 0.01 5.44 ± 0.05 0.83 ± 0.02 0.81 ± 0.06 0.79 ± 0.06 0.78 ± 0.06 0.71 ± 0.06
CMC 7.95 ± 0.02 7.36 ± 0.06 7.10 ± 0.07 6.97 ± 0.03 0.78 ± 0.01 0.75 ± 0.05 0.64 ± 0.06 0.59 ± 0.06
SA 8.09 ± 0.05 7.81 ± 0.08 7.27 ± 0.04 7.05 ± 0.09 0.76 ± 0.01 0.71 ± 0.07 0.61 ± 0.06 0.60 ± 0.05
a
Not detected.

and SA, which exhibit linear chain architectures. Moreover, Ln is the
precursor of humic substances and possesses a similar structure to
humic acids (HAs), which are facile in combining with soil colloids
to form insoluble complexes with metal in soils (Senesi et al., 2007).
Ln's aromatic structure was more difficult to be decomposed and
degraded by soil organisms and much more stable than the struc-
tures of CMC and SA (KogeleKnabner, 2002; Lewis and Yamamoto,
1990). However, the different efficiencies of the three natural
polymers in the immobilization of Pb and Cd may have resulted
from the differences in oxygen-containing functional groups with
various affinities toward different metals (Speight, 2005). Ln con-
tains carboxyl, lactonic, and phenolic hydroxyl groups, whereas
CMC and SA mainly consist of carboxymethyl and hydroxyl groups.
Interestingly, the adsorption capacities of CMC and SA for Pb and
Cd were both higher than that of Ln in aqueous solutions from
Supplementary Materials Fig. S2, but the immobilization effects of
Ln on those two metals in soils were more evident than the other
two natural polymers. It may be due to two facts. One was different
measured pH levels at soils and at solutions causing various
external environments. The other was their far different molecular
structures resulting in various immobilization efficiencies in
different media.
Pb and Cd coexist in the soils; hence, aPb
Cd
, which is the selectivity
factor of adsorbate Pb over adsorbate Cd, was introduced and
calculated using Eq. (2) to study their competitive immobilization
behavior. The results are also listed in Table 3.
Kd ðPbÞ
aPb
Cd ¼ (2)
Kd ðCdÞ
Most of the a values of Ln were higher than 1.0 but those of CMC
352 X. Tao et al. / Environmental Pollution 222 (2017) 348e355

Table 3
The Kd values of targeted metals by TCLP extraction and their exchangeable form by sequential extraction as well as their aPb
Cd
.

By TCLP extraction

Kd (Pb) Kd (Cd) aPb

Cd

1% 2% 5% 10% 1% 2% 5% 10% 1% 2% 5% 10%

Soil1 Ln 0.29 0.65 0.65 0.71 0.045 0.20 0.24 0.28 6.30 3.19 2.68 2.52
CMC 0.11 0.14 0.16 0.21 0.091 0.35 0.46 0.46 1.17 0.40 0.35 0.45
SA 0.081 0.10 0.14 0.20 0.023 0.27 0.35 0.39 3.55 0.37 0.41 0.51
Soil2 Ln 0.42 0.72 1.00 1.00 0.025 0.061 0.086 0.12 16.54 11.68 11.69 8.22
CMC 0.025 0.060 0.15 0.17 0.013 0.086 0.098 0.16 1.87 0.71 1.57 1.08
SA 0.036 0.055 0.13 0.14 0.013 0.061 0.12 0.15 2.69 0.89 1.07 0.94

By sequential extraction

Kd (Pb) Kd (Cd) aPb

Cd

1% 2% 5% 10% 1% 2% 5% 10% 1% 2% 5% 10%

Soil1 Ln 0.18 0.55 0.74 0.77 0.42 0.52 0.56 0.58 0.42 1.06 1.32 1.32
CMC 0.16 0.21 0.36 0.39 0.48 0.53 0.59 0.64 0.34 0.39 0.39 0.4
SA 0.13 0.18 0.25 0.35 0.49 0.51 0.55 0.62 0.13 0.85 0.79 0.83
Soil2 Ln 0.053 0.25 0.26 0.34 0.024 0.048 0.060 0.14 2.18 5.14 4.24 2.32
CMC 0.029 0.10 0.13 0.15 0.060 0.096 0.23 0.29 0.49 1.05 0.58 0.52
SA 0.012 0.046 0.11 0.14 0.084 0.14 0.18 0.28 0.14 0.32 0.42 0.50

and SA were near to or slightly lower than 1.0 (Table 3). This finding efficiencies for Pb and Cd, which can be ascribed to the different
suggests that Ln exhibited selective adsorption for Pb, but the oxygen-containing functional groups and various affinities to
selectivity of CMC and SA was not evident (Table 3). The three different metals (Speight, 2005).
employed natural polymers exhibited different immobilization

Fig. 2. Changes of metal fractionations (Pb and Cd) in different soil samples after treated by various contents of different organic stabilizers, through sequential extraction. (CK:
untreated soil; the contents of stabilizers in Ln1-Ln4, CMC1-CMC4, and SA1-SA4 are 1, 2, 5, and 10 wt% respectively.)
 X. Tao et al. / Environmental Pollution 222 (2017) 348e355
3.2. Sequential extraction
The sequential chemical extraction procedures can help in
assessing the potential mobility and solubility of metals in
contaminated soils. Hence, the exchangeable Pb and Cd concen-
trations of both treated and untreated soils were further deter-
mined by sequential extraction (Table 2). The exchangeable metal
form in soils is usually the most mobile and toxic form. In addition,
besides the analyses of the exchangeable (E) form, the re-
distributions of metal fractionation in the soils were also evaluated
in the carbonate (C), FeeMn oxide (FM), organic (O), and residual
forms (R) (Fig. 2). In Table 2 and Fig. 2, the trends of Pb and Cd
reduction in exchangeable form by sequential extraction were
much similar to those by TCLP extraction. The metal concentrations
in the exchangeable form after treatment using natural polymers
were generally lower than those of the untreated counterpart in all
the measured soil samples. The higher doses of stabilizers were
usually more effective in reducing the concentrations of the mobile
metals (Table 2 and Supplementary Materials Fig. S3).
Similarly, on the basis of Table 2, the immobilization efficiency
and selectivity of the three stabilizers were further evaluated
through the Kds of two metals and aPb Cd
calculated using Eqs. (1) and
(2) (Table 3). Accordingly, Ln showed higher immobilization effi-
ciency and selectivity for Pb than the other two stabilizers. How-
ever, CMC and SA exerted better effects than those of Ln in the
immobilization of Cd. These results may be attributed to the dif-
ferences in chain architecture of the natural polymers and the
dissimilar affinities of the oxygen-containing functional groups on
the different stabilizers for the two metals.
In addition, the exchangeable Pb and Cd concentrations
Fig. 3. The concentration of targeted metals in two different soil samples after amendment with various stabilization agents by TCLP extraction as a function of the sum of the
concentrations of respective exchangeable (E) and carbonate (C) forms obtained by sequential extraction.
353
detected by sequential extraction were mostly lower than those
detected by TCLP extraction in Table 2 because of the difference in
extraction efficiencies and conditions. The exchangeable form of
metals was leached by MgCl2 at pH 7 in sequential extraction.
Meanwhile, TCLP extraction adopted acetic acid as the extracting
agent and thus maintained the entire extraction process under a
lower pH level (Liu and Gonzalez, 1999). Consequently, partial
carbonate-bound forms of the metals were also extracted.
On the basis of Fig. 2 and Table 2, the TCLP concentrations of Pb
and Cd as a function of the sum of the concentrations of respective
exchangeable and carbonate forms obtained by sequential extrac-
tion are shown in Fig. 3. Most of the TCLP concentrations of the two
metals were linearly proportionate to the total concentration of the
respective exchangeable and carbonate forms (Fig. 3 and Supple-
mentary Materials Table S3), of which the correlation coefficients
(R2s) were fairly close to 1.0. These findings suggest that the
carbonate-bound metals were unstable in acidic media and exert
potential environmental risk.
Meanwhile, the organic-bound form of the metals in the soils
increased in amount with the increase in stabilizer dose because of
the organic polymeric stabilizers used in the study (Fig. 2 and
Supplementary Materials Fig. S3). Moreover, the TCLP concentra-
tions of Pb and Cd in two different soil samples after amendment
with various stabilization agents as a function of the concentration
of respective organic form are shown in Fig. 4. The group obviously
displays an inversely proportional relationship in Fig. 4 and Sup-
plementary Materials Table S3. This result indicated that the
organic form of metals was sufficiently stable to resist the leaching
owing to the effective chelating effects between the oxygen-
containing groups on natural polymers and metals (Conte et al.,
354 X. Tao et al. / Environmental Pollution 222 (2017) 348e355
Fig. 4. The concentration of targeted metals in two different soil samples after amendment with various stabilization agents by TCLP extraction as a function of the concentration of
respective organic (O) form obtained by sequential extraction.
2005; Karlsson et al., 2007).
Besides, in accordance with Fig. 2 showing the redistribution of
the carbonate-bound and FeeMn-bound forms of Pb and Cd, some
fluctuations were noted during the incubation periods. The appli-
cation of stabilizers caused slight changes in soil pH levels, which
may result in the fluctuation of the carbonate-bound form of Pb and
Cd in soils (Fig. 1). Meanwhile, the addition of distilled water during
incubation time would change the redox potentials of the soils and
pose some effects on their FeeMn bound form in soils (Husson,
2013). Meanwhile, the residual forms of metals therefore
increased implying the formation of some more stable substances
(Conte et al., 2005).
At last, it should be pointed out that the sequential extraction
methods also have experimental limitations although they can
provide comparative information relating to the relative contribu-
tion of the target compound and the metal mobility. The concept of
these fractionation methods is based on the idea that each
extracting solution is highly selective for a particular phase of the
solid, thus inducing the solubilization of associated elements.
However, this selectivity cannot be expected for most of the pro-
cedures due to the fact that a mass transfer and redistribution of
metal may occur between compartments during extraction and
separation (Bacon and Davidson, 2008; Dermont et al., 2008b;
Gleyzes et al., 2002; Templeton et al., 2000). Thus, the sequential
chemical extraction procedures determine neither specific chemi-
cal forms of a metal nor the speciation of metals in the solid or
liquid phases of soil, but metal fractionation according to bonding
with specific soil substrates (Bacon and Davidson, 2008; Dermont
et al., 2008b; Gleyzes et al., 2002; Templeton et al., 2000). The
aforementioned deficiencies should be also taken into
consideration when the sequential extraction procedures were
employed to assess the chemical forms of metals in the contami-
nated sediment and control the sediment treatment processes.
4. Conclusions
Three natural polymer amendments used herein were all
effective in immobilizing Pb and Cd in contaminated soils because
of their high affinity and efficient chelating effects between the
abundant oxygen-containing functional groups on stabilizers and
metals. The addition of Ln, CMC, and SA increased soil pH levels by
about 0.078e0.535 and decreased TCLP concentrations of Pb and Cd
by about 5.46%e71.1% and 4.25%e49.6%, respectively, compared
with those of the untreated soils. Among the three organic stabi-
lizers, Ln was the most efficient in remediating the Pb- and Cd-
contaminated soils with higher immobilization and better dura-
tion. These results were ascribed to Ln's distinct three-dimensional
molecular structure and various oxygen-containing functional
groups. Moreover, Ln demonstrated greater preferential immobi-
lization of Pb over Cd than the other two stabilizers in soils, which
may be due to the variation in affinity of the different oxygen-
containing functional groups on the natural polymers toward
different metals. In brief, the natural polymers, especially Ln, hold
good application potentials in the remediation of metal-
contaminated soils.
Acknowledgments
This work was supported by the Natural Science Foundation of
China (grant no. 51378250), the Natural Science Foundation of
 X. Tao et al. / Environmental Pollution 222 (2017) 348e355 355

Jiangsu Province (grant no. BK20161405), and Six Talent Peaks in soil using amendments-a review. Waste Manage 28 (1), 215e225.
Kuo, C.K., Ma, P.X., 2001. Ionically crosslinked alginate hydrogels as scaffolds for
Project in Jiangsu Province of China (grant no. 2015-JNHB-003).
tissue engineering: Part 1. Structure, gelation rate and mechanical properties.
Thanks very much for Prof. Rong JI's helpful suggestion and Biomaterials 22, 511e521.
discussion. Lageman, R., Clarke, R.L., Pool, W., 2005. Electro-reclamation, a versatile soil
remediation solution. Eng. Geol. 77 (3e4), 191e201.
Lagergren, S., 1898. About the theory of so-called adsorption of soluble substances.
Appendix A. Supplementary data Kung. Sven. Veten. Hand 24, 1e39.
Langmuir, I., 1918. The adsorption of gases on plane surfaces of glass, mica and
Supplementary data related to this article can be found at http:// platinum. J. Amer. Chem. Soc. 40, 1361e1403.
Lee, S.H., Lee, J.S., Choi, Y.J., Kim, J.G., 2009. In situ stabilization of cadmium-, lead-,
dx.doi.org/10.1016/j.envpol.2016.12.028. and zinc-contaminated soil using various amendments. Chemosphere 77 (8),
1069e1075.
References Lewis, N.G., Yamamoto, E., 1990. Lignin-occurrence, biogenesis and biodegradation.
Annu. Rev. Plant Physiol. 41, 455e496.
Li, X.D., Poon, C.S., Sun, H., Lo, I.M.C., Kirk, D.W., 2001. Heavy metal speciation and
Bacon, J.R., Davidson, C.M., 2008. Is there a future for sequential chemical extrac-
leaching behaviors in cement based solidified/stabilized waste materials.
tion? Analyst 133 (1), 25e46.
J. Hazard. Mater 82 (3), 215e230.
Bao, S., 1981. In: Li, G., Yang, G. (Eds.), Soil Agro-chemistrical Analysis, first ed. China
Li, Z.Y., Ma, Z.W., van der Kuijp, T.J., Yuan, Z.W., Huang, L., 2014. A review of soil
Agricultural Press, Beijing, pp. 8e31.
heavy metal pollution from mines in China: pollution and health risk assess-
Battaglia, A., Calace, N., Nardi, E., Petronio, B.M., Pietroletti, M., 2007. Reduction of
ment. Sci. Total Environ. 468e469, 843e853.
Pb and Zn bioavailable forms in metal polluted soils due to paper mill sludge
Liao, X.Y., Li, Y., Yan, X.L., 2016. Removal of heavy metals and arsenic from a co-
addition-Effects on Pb and Zn transferability to barley. Bioresour. Technol. 98
contaminated soil by sieving combined with washing process. J. Environ. Sci.
(16), 2993e2999.
China 41, 202e210.
Beesley, L., Moreno-Jimenez, E., Gomez-Eyles, J.L., Harris, E., Robinson, B., Sizmur, T.,
Liu, A.G., Gonzalez, R.D., 1999. Adsorption/desorption in a system consisting of
2011. A review of biochars' potential role in the remediation, revegetation and
humic acid, heavy metals, and clay minerals. J. Colloid Interf. Sci. 218, 225e232.
restoration of contaminated soils. Environ. Pollut. 159 (12), 3269e3282.
Ma, L.Q., Komar, K.M., Tu, C., Zhang, W.H., Cai, Y., Kennelley, E.D., 2001. A fern that
Boehm, H.P., 1994. Some aspects of the surface-chemistry of carbon-blacks and
hyperaccumulates arsenic. Nature 409 (6820), 579.
other carbons. Carbon 32 (5), 759e769.
Maini, G., Sharman, A.K., Sunderland, G., Knowles, C.J., Jackman, S.A., 2000. An in-
Bolan, N., Kunhikrishnan, A., Thangarajan, R., Kumpiene, J., Park, J., Makino, T.,
tegrated method incorporating sulfur-oxidizing bacteria and electrokinetics to
Kirkham, M.B., Scheckel, K., 2014. Remediation of heavy metal(loid)s contami-
enhance removal of copper from contaminated soil. Environ. Sci. Technol. 34
nated soils-to mobilize or to immobilize? J. Hazard. Mater 266, 141e166.
(6), 1081e1087.
Calace, N., Campisi, T., Iacondini, A., Leoni, M., Petronio, B.M., Pietroletti, M., 2005.
Mallampati, S.R., Mitoma, Y., Okuda, T., Sakita, S., Kakeda, M., 2012. Enhanced heavy
Metal-contaminated soil remediation by means of paper mill sludges addition:
metal immobilization in soil by grinding with addition of nanometallic Ca/CaO
chemical and ecotoxicological evaluation. Environ. Pollut. 136 (3), 485e492.
dispersion mixture. Chemosphere 89 (6), 717e723.
Cheng, S., 2003. Heavy metal pollution in China: origin, pattern and control. En-
Mignardi, S., Corami, A., Ferrini, V., 2012. Evaluation of the effectiveness of phos-
viron. Sci. Pollut. R. 10 (3), 192e198.
phate treatment for the remediation of mine waste soils contaminated with Cd,
Conte, P., Agretto, A., Spaccini, R., Piccolo, A., 2005. Soil remediation: humic acids as
Cu, Pb, and Zn. Chemosphere 86 (4), 354e360.
natural surfactants in the washings of highly contaminated soils. Environ.
Park, J.H., Lamb, D., Paneerselvam, P., Choppala, G., Bolan, N., Chung, J.W., 2011. Role
Pollut. 135 (3), 515e522.
of organic amendments on enhanced bioremediation of heavy metal(loid)
Dahrazma, B., Mulligan, C.N., 2007. Investigation of the removal of heavy metals
contaminated soils. J. Hazard. Mater 185 (2e3), 549e574.
from sediments using rhamnolipid in a continuous flow configuration. Che-
Perkerson, F.S., Reeves, W.A., Tripp, V.W., 1960. Some studies on the cross-linking of
mosphere 69 (5), 705e711.
cotton cellulose. Part I: chemical and physical aspects. Text. Res. J. 30, 944e955.
D'Amore, J.J., Al-Abed, S.R., Scheckel, K.G., Ryan, J.A., 2005. Methods for speciation of
Querol, X., Alastuey, A., Moreno, N., Alvarez-Ayuso, E., Garcia-Sanchez, A., Cama, J.,
metals in soils: a review. J. Environ. Qual. 34, 1707e1745.
Ayora, C., Simon, M., 2006. Immobilization of heavy metals in polluted soils by
Dermont, G., Bergeron, M., Mercier, G., Richer-Lafleche, M., 2008a. Soil washing for
the addition of zeolitic material synthesized from coal fly ash. Chemosphere 62
metal removal: a review of physical/chemical technologies and field applica-
(2), 171e180.
tions. J. Hazard. Mater 152 (1), 1e31.
Rahimi, M., Farhadi, R., Mehdizadeh, R., 2013. Phytoremediation: using plants to
Dermont, G., Bergeron, M., Mercier, G., Richer-Lafle che, M., 2008b. Metal-contam-
clean up contaminated soils with heavy metals. Int. J.Agr. Res. 3 (1), 148e152.
inated soils: remediation practices and treatment technologies. Pract. Period.
Senesi, N., Plaza, C., Brunetti, G., Polo, A., 2007. A comparative survey of recent
Hazard. Toxic Radioact. Waste Manage 12 (3), 188e209.
results on humic-like fractions in organic amendments and effects on native
Freundlich, H., 1906. Concerning adsorption in solutions. Z. Phys. Chem. 57 (4),
soil humic substances. Soil Biol. Biochem. 39 (6), 1244e1262.
385e470.
Shao, X., Huang, B., Zhao, Y., Sun, W., Gu, Z., Qian, W., 2014. Impacts of human ac-
GB15618, 2008. China. In: Environmental Quality Standards for Soils.
tivities and sampling strategies on soil heavy metal distribution in a rapidly
Gleyzes, C., Tellier, S., Astruc, M., 2002. Fractionation studies of trace elements in
developing region of China. Ecotox. Environ. Safe 104, 1e8.
contaminated soils and sediments: a review of sequential extraction proced-
Speight, J.G., 2005. Lange's Handbook of Chemistry, sixth ed. McGraw-Hill, New
ures. Trend Anal. Chem. 21 (6e7), 451e467.
York.
Gonza
lez-Nún ~ ez, R., Alba, M.D., Orta, M.M., Vidal, M., Rigol, A., 2011. Remediation of
Sun, Y.B., Sun, G.H., Xu, Y.M., Wang, L., Lin, D.S., Liang, X.F., Shi, X., 2012. In situ
metal-contaminated soils with the addition of materials e Part I: character-
stabilization remediation of cadmium contaminated soils of wastewater irri-
ization and viability studies for the selection of non-hazardous waste materials
gation region using sepiolite. J. Environ. Sci. China 24 (10), 1799e1805.
and silicates. Chemosphere 85 (9), 1511e1517.
Templeton, D.M., Cornelis, R., Danielsson, L.G., Muntau, H., Van Leeuwen, H.P.,
Guo, G.L., Zhou, Q.X., Ma, L.Q., 2006. Availability and assessment of fixing additives
Lobinski, R., 2000. Guidelines for terms related to chemical speciation and
for the in situ remediation of heavy metal contaminated soils: a review. Envi-
fractionation of elements. Definitions, structural aspects, and methodological
ron. Monit. Assess. 116 (1e3), 513e528.
approaches (IUPAC Recommendations 2000). Pure Appl. Chem. 72 (8),
Harter, R.D., 1983. Effect of soil-pH on adsorption of lead, copper, zinc, and nickel.
1453e1470.
Soil. Sci. Soc. Am. J. 47 (1), 47e51.
Tessier, A., Campbell, P.G.C., Bisson, M., 1979. Sequential extraction procedure for
Ho, Y.S., McKay, G., 1998. Sorption of dye from aqueous solution by peat. Chem. Eng.
the speciation of particulate trace-metals. Anal. Chem. 51 (7), 844e851.
J. 70 (2), 115e124.
Tica, D., Udovic, M., Lestan, D., 2011. Immobilization of potentially toxic metals
Husson, O., 2013. Redox potential (Eh) and pH as drivers of soil/plant/microor-
using different soil amendments. Chemosphere 85 (4), 577e583.
ganism systems: a transdisciplinary overview pointing to integrative opportu-
USEPA, 2007. The Use of Soil Amendments for Remediation, Revitalization and
nities for agronomy. Plant Soil 362 (1e2), 389e417.
Reuse. EPA 542-R-07e013.
Jarup, L., 2003. Hazards of heavy metal contamination. Brit. Med. Bull. 68, 167e182.
Vaca-Paulin, R., Esteller-Alberich, M.V., Lugo-de la Fuente, J., Zavaleta-Mancera, H.A.,
Jiang, T.D., 2009. Lignin, second ed. Chemical Industry Press, Beijing.
2006. Effect of sewage sludge or compost on the sorption and distribution of
Karlsson, T.R., Elgh-Dalgren, K., Bjorn, E., Skyllberg, U., 2007. Complexation of cad-
copper and cadmium in soil. Waste Manage 26 (1), 71e81.
mium to sulfur and oxygen functional groups in an organic soil. Geochim.
Wei, B., Yang, L., 2010. A review of metal contaminations in urban soils, urban road
Cosmochim. Ac. 71, 604e614.
dusts and agricultural soils from China. Microchem. J. 94 (2), 99e107.
Kogel-Knabner, I., 2002. The macromolecular organic composition of plant and
Zhou, Y.F., Haynes, R.J., 2010. Sorption of metals by inorganic and organic compo-
microbial residues as inputs to soil organic matter. Soil Biol. Biochem. 34 (2),
nents of solid wastes: significance to use of wastes as low-cost adsorbents and
139e162.
immobilizing agents. Crit. Rev. Env. Sci. Technol. 40 (11), 909e977.
Kumpiene, J., Lagerkvist, A., Maurice, C., 2008. Stabilization of As, Cr, Cu, Pb and Zn