J Crit Rev, Vol 1, Issue 1, 36-39 Review Article



Centre for Nanobiotechnology, VIT University, Vellore 632014, Tamil Nadu, India.
Email: iniyasahamed@vit.ac.in

Received: 21 Jul 2014 Revised and Accepted: 14 Sep 2014


Nanotechnology is based on the idea that, by engineering the size and shape of materials at the scale of atoms, i. e. nanometers (nm), distinct optical, electronic, or magnetic properties can be tuned to produce novel properties of commercial value. However, there is an obvious concern that such novel properties may also lead to novel behavior when interacting with biological organisms, and thus to potentially novel toxic effects. Nano zero-valent iron particle has been used for site remediation by creating permeable reactive barriers (PRBs) by filling trenches with ZVI designed to allow groundwater to pass through while “filtering” out the contaminants. Research indicates that using the nanoscale ZVI in place of the macroscale ZVI will accomplish the same remedial work more efficiently and with less cost. The full acceptance of NZVIs as a remediation agent depends on several issues. One of the most important factors relates to the fate, impact and toxicity of these nanomaterials on the ecosystems to which they are applied.

Keywords: Nanotechnology, Nano zero-valent iron, Ecotoxicity.


In recent decades, the use of nanomaterials has become increasingly significant in industrial processes, consumer and medical products [1]. And more recently, in environmental remediation [2]. This has led to the introduction of significant amounts of distinct types of nanomaterials into all the environmental categories: soils, aquatic systems and air. In soils, nanoparticles can be introduced either directly, through fertilizers and products used for plant protection or liquid suspensions used in contaminated sites, or indirectly, through the land application of sludge or biosolids. The presence of nanoparticles in aquatic systems is mainly due to the disposal of wastewater treatment plant effluents, industrial discharges, and surface runoff from soils. Volcanic eruptions, combustion processes and industrial emissions are some of the sources of nanoparticles release in the air [3].

After the introduction of nanoparticles into the environment, the particles undergo several changes involving biological, physical and chemical processes, which make it difficult to quantify their prevalence and evaluate their degree of ecotoxicity. In particular, these changes include chemical interactions (e. g. redox reactions) and agglomeration effects. The type and extent of these processes depend on the properties of both the nanoparticles and the receiving medium [1]. For example, in aqueous systems, hardness, biochemical oxygen demand, pH, alkalinity and organic matter content are some of the parameters that influence the behavior of the nanoparticles.

Difficulty relates to the quantification of trace amounts of nanoparticles in the environment. Knowledge of this subject remains scarce mostly because there are neither any specific standardized methods nor protocols nor any certified reference materials for the testing of nanomaterials [4, 5]. Nevertheless, the scientific community is trying to adopt the best methodologies for conducting such studies. All these facts contribute to the growing concerns about the fate and effects of these materials in the environment. This encourages and puts pressure on the scientific community to answer these issues and to evaluate the real impact of nanomaterial usage, nanomaterial ecotoxicity and the need for more information on the proper handling of these materials in order to prevent environmental and human health effects after long-term exposure [4]. Macroscale zero-valent iron particle (ZVI) has long been recognized as an excellent electron donor with a tendency to release electrons in aquatic environments regardless of its particle size [5]. ZVI has been used for site remediation since the early 1990s by creating permeable reactive barriers (PRBs) by filling trenches with ZVI designed to allow groundwater to pass through while “filtering” out the contaminants [6, 7]. Research indicates that using the nanoscale ZVI (NZVI) in place of the macroscale ZVI will accomplish the same remedial work more efficiently and with less cost. A benefit of using NZVI is being able to inject it directly into a contaminated aquifer, which avoids the need to dig a trench for installation of the PRB. Using this injection technique is believed to be faster and more effective for groundwater treatment than either pump-and-treat or PRB methods [6].

ZVI, or elemental iron, is a moderate reducing reagent that can react with dissolved oxygen and water resulting in electrochemical/corrosion reactions that oxidize the iron [8]. As shown below, the Fe0 becomes oxidized to ferrous iron (Fe2+) ions and the electrons that are released become available to reduce other compounds. This process can be accelerated or inhibited by changing the solution chemistry and/or solid composition by adding coatings or catalysts [8].

2Fe0(s) + 4H+ (aq) + O2 (aq) → 2Fe2+(aq) + 2H2O(l) (1)

Fe0(s) + 2H2O (aq) → Fe2+ (aq) + H2 (g) + 2OH-(aq) (2)

The chemistry behind NZVI is much the same as with macroscale ZVI. NZVI is highly reducing and generates reactive oxygen species (ROS) through Fenton chemistry. In aqueous environments, NZVI oxidizes over time (i. e., ages) to iron oxides such as magnetite, maghemite, lepidocrocite, and goethite [9, 10].

Research has shown the NZVI can react not only with dissolved oxygen and water, but also with a variety of environmental contaminants, and may prove to be a more effective and less costly remediation alternative. NZVI can remove contaminants through reduction or adsorption. The treatment of chlorinated solvents, for example, primarily uses reduction, and the removal of arsenic is done by adsorption to iron oxides and hydroxides formed during oxidation. An example that has received a lot of attention is the process of degrading trichloroethene (TCE) and perchloroethylene (PCE), TCE and PCE were used historically as industrial cleaning and degreasing solvents, as paint removal solvents, and as dry-cleaning solvents [11, 12]. Chlorinated solvents can be degraded through reductive dechlorination or beta elimination. Under the reducing conditions presented by the introduction of NZVI in groundwater, reductive dechlorination follows the pathway of PCE → TCE → DCE → VC → ethane, where DCE is dichloroethene and VC is vinyl chloride [7].

Although NZVI particles can exist naturally, remediation techniques generally use engineered nanoparticles. NZVI can be created in a variety of different ways. Researchers discovered three distinct methods for producing NZVI through the reduction and precipitation of ZVI from an aqueous solution using sodium borohydride for the chemical reduction of ferrous (Fe2+) or ferric (Fe3+) ions. NZVI also can be produced by hydrogen reduction of iron oxides or hydroxides.

The following equations illustrate some of the reactions that can take place to produce NZVI [14, 15]:

Fe (H2O)6 3+ + 3BH4 - + 3H2O → Fe0↓ +3B(OH)3 + 10.5H2 (3)

4Fe3+(aq) + 3BH-4(aq) + 9H20(l) → 4Fe0(s) + 3H2BO-3(aq) + 12H+(aq) + 6H2(g) (4)

2Fe2+ (aq) + BH-4(aq) + 3 H20 (l) → 2Fe0(s) + H2BO-3(aq) + 4H+(aq) + 2H2(g) (4)

It is important to know how the nanoparticles are produced. Even particles with the same chemical composition will have different reactivity, mobility and active life due to the process or vendor used to obtain the nanoparticles [13]. Contamination also can be an issue for reactivity. The greater the percentage of ZVI present in the resulting nanoscale powder, the more reactive it will be. When Fe2+ or Fe3+ ions is present in the NZVI powder it will be less reactive due to the decreased availability of electrons [16].

There are three main forms of nanoscale iron that can be used during remediation efforts – NZVI, bimetallic nanoscale particles (BNP) and emulsified zero-valent iron (EZVI). BNPs are particles of NZVI that have been coated with a catalyst, such as platinum, gold, nickel, or palladium, to increase mobility and enhance reduction reaction. EZVI are NZVI particles that have been coated with a membrane made from biodegradable oil and water to facilitate the treatment of chlorinated hydrocarbons by making the particles more hydrophobic. This allows the particles to mix directly with dense non-aqueous phase liquids (DNAPL), such as TCE, to increase mass transfer between the DNAPL and the NZVI through the emulsion membrane.

According to the Environmental Protection Agency (EPA), NZVI is the most common nanoparticles that has been tested and field applied. It constitute one of the most common materials used in nano-remediation because of their high superficial area and reactivity with distinct contaminants such as metals, halogenated hydrocarbons [18]. polychlorinated biphenyls (PCBs) [19]. and pharmaceutical products [20].

The high efficiencies obtained in recent tests in the laboratory and in pilot studies indicate that the use of NZVIs is extremely promising in terms of environmental remediation [20-26]. However, as with the other nanomaterials, several concerns are being raised about the impact and ecotoxicity of NZVIs.

This is largely due to the formation of iron oxides, which are already present in the ground as rust, during remediation [27]. The toxicity of iron is based on its ability to catalyze the formation of hydroxyl radicals (OH.) from superoxide (O2.-) and hydrogen peroxide (H2O2). Free radicals are highly reactive, unstable molecules that are in need of an additional electron for stabilization. Because of this, they can “affect antioxidant enzymatic activities, per oxidation of membrane lipids, modification of nucleic acids, and eventually cause cell death and tissue injury” [27].

Iron toxicity studies have primarily focused on Fe2+ and its oxides and little is known about the toxicity specific to NZVI or macroscale ZVI. However, ZVI produces Fe2+ and iron oxides through oxidation and ZVI/NZVI can produce free radicals through this transformation process.

Toxicity performance of NZVIs against soil and aquatic organisms

Bacteria, along with algae, are at the bottom of the aquatic food chain, being the food of aquatic crustaceans such as Daphnia, which are in turn consumed by fish. In vitro tests have shown that NZVIs are bactericidal to certain aqueous cultures of Escherichia coli or Bacillus nealsonii [29-31]. In contaminated environments, the native microbial consortia are generally already inhibited by the presence of significant concentrations of the contaminants.

Certain studies in environmental matrices showed opposite results. Fajardo et al. observed that NZVIs had a reduced impact on microbial cellular viability and on biological activity in soils of Klebsiella planticola and Bacillus nealsonii and concluded that the ecotoxicity of NZVIs could be highly dose- and species dependent [29]. Kirschling et al. observed that NZVIs had no effect on the bacterial abundance in the soil and that the bacterial populations increased when the NZVIs were coated with a biodegradable poly aspartate [32]. This could indicate that the use of coated NZVIs may reduce their toxicity. On the other hand, Barnes et al. [33] observed a negative impact of NZVIs on the capacity of an indigenous dechlorinating bacterial community to degrade trichloroethylene (TCE). This impact was dose dependent: the biological degradation rate started to decrease at NZVI concentrations above 0.01 g/L and ceased at concentrations above 0.3 g/L.

Studies indicate that there is a possible impact of NZVIs on bacterial communities and that this impact is dose dependent. Therefore, NZVIs can be applied to soils, but their dosage should not exceed the level that is detrimental to bacteria. On the other hand, the use of coated NZVIs can enhance environmental remediation. However, it is not certain that these coatings can allow for the use of higher NZVI dosages without causing a negative effect on bacteria. This is a field of research that should be explored further in order to widen the applicability of NZVIs.

Aquatic invertebrates are commonly affected by most of the contaminants released into the environment. This is one of the reasons that these organisms are important and appropriate for ecotoxicity tests [34]. Behavior and bioavailability of nanoparticles in aquatic environments, invertebrate testing, used to increase knowledge of their toxicology. Daphnia magna is used in the majority of tests to evaluate the ecotoxicity of nanoparticles [35].

Several studies show that Daphia magna is generally very sensitive to the presence of distinct nanoparticles; continuous exposure leads to the immobilization or death of the organisms [36]. However, only a few works have focused on the ecotoxicity of NZVIs. Marsalek et al. Studied the possible application of NZVIs to destroy and prevent, in a simple and environmentally benign way, the formation of cyanobacterial water blooms. They observed an NZVI EC50 of 50 mg/L against cyanobacteria, while for Daphnia magna they observed an EC50 higher than 1000 mg/L [37]. Keller et al. Registered that Daphnia magna survival was drastically influenced by commercial NZVIs (Nanofer 25S and Nanofer STAR) [38].

These few studies indicate that NZVIs significantly affect the Daphnia magna communities and can even lead to their death. Considering that the use of NZVIs for environmental remediation is mainly focused on contaminated waters, these results are very important and reinforce the need for more detailed and structured studies. These studies consider the impact of NZVIs on Daphnia magna in the absence of contaminants. However, the NZVIs are applied to contaminated environments, and therefore future research should evaluate the impact of contaminants on Daphnia magna/other speciesas well as the additional impact of NZVIs.

Earthworms are common soil organisms that play an important and distinct role in the soil ecosystem and for this reason; they are used as test organisms in soil ecotoxicity studies [39]. And to assess the bioavailability of contaminants in soils [40]. On account of the limitations in forming reliable conclusions about the validity of these tests, NZVIs are generally only applied to specific situations in which contaminations have occurred and where evaluation is required. However, some studies have examined the ecotoxicity of different nanoparticles {e. g. Aluminum oxide [41]. Silver [42]. or titanium oxide [43]. in soils, but, as far as is known; only one study has focused on the ecotoxicity of NZVIs in earthworms.

The eco-toxicological effects of NZVIs coated with carboxymethyl cellulose on Eisenia fetida and Lumbricus rubellus. This work proved the negative impact of NZVIs on both of these earthworm species, affecting reproduction when the NZVI concentration reached 100 mg/kg and leading to decreased weight and an increased mortality rate in concentrations above 500 mg/kg [44].

The present insufficiencies of information delay a supported assessment of the impact of NZVIs on terrestrial organisms. However, research indicates that, above specific NZVI concentrations, the organisms’ reproduction, weight and mortality rates are affected.

It is clear that further research is needed and such research should evaluate the relative impact of NZVI application on contaminated environments. Research to be carried out in other types of earthworms to assess at which concentrations, the organism’s reproduction, weight and mortality rates are affected.

Lactuca sativa, the common lettuce, is probably the plant that is more often used for germination test on account of its high sensitivity to distinct contaminants. Other plants are also used, such as cabbage (Brassica oleracea L.), corn (Zea mays) or soybean (Glycine max), but there is still no consensus on the most appropriate plant for such a test [45]. These tests are commonly used for soils contaminated with distinct contaminants such as metals [46]. Petroleum hydrocarbons [47] or pharmaceutical products [48]. A few studies have been performed with nanomaterials [49, 50]. Barrena et al. studied the toxicity of gold, silver and iron oxide (Fe3O4) nanoparticles on cucumber (Cucumis sativus) and lettuce (Lactuca sativa) [49].

Ravindran et al. performed germination tests with Lycopersicum esculentum and Zea mays to evaluate the ecotoxicity of silver nanoparticles and silver ions.

This study carried out in our laboratory, showed a higher toxic effect with silver nanoparticles than with silver ions; however, when the nanoparticles were supplemented with bovine serum albumin, there was a reduction in adverse effects [50].

The ecotoxicity of NZVIs and three types of silver nanoparticles in germination tests with ryegrass, barley and flax was conducted. In aquatic systems, inhibitory effects were observed for NZVI concentrations of 250 mg/L, while concentrations of 1000–2000 mg/L completely inhibited germination [51]. The tolerance of Panicum maximum (purple guinea grass) and Helianthus annuus (common sunflower) in a TNT-contaminated soil and in an NZVI-contaminated soil was studied. Panicum maximum showed more tolerance than Helianthus annuus to the presence of NZVIs [52].

This type of test not only indicates the impact of NZVIs on the germination process; it can also provide information on the uptake of NZVIs by the plant’s roots and leaves. This knowledge allows a more complete and thorough evaluation of the impact of NZVIs on plants. Nevertheless, there is some evidence that indicates that the germination of some plants is affected by the presence of NZVIs. Therefore, in order to protect superficial plants, NZVI suspensions supposed to be applied in soils via deep slurry injections.

Fig. 1: Diseases associated to nanoparticles exposure

Source: Ecotoxicity of nanoscale zero-valent iron particles – a review. http: //www. visaemdebate. incqs. fiocruz. br/


As per the available literature, not sufficient work was carried out in ecotoxicity behavior of NZVIs with special reference to India. Few studies have been completed; several research groups are currently in the process of analyzing data pertaining specifically to the toxicity of NZVI. The full acceptance of NZVIs as a remediation agent depends on several issues. One of the most important factors relates to the fate and impact of these nanomaterials on the ecosystems to which they are applied. The existing literatures are clearly insufficient. Nevertheless, the majority of the studies point toward the toxic effects of NZVIs on all the tested organisms.


The author would like to thank the honorable chancellor Dr. G. Viswanathan, the vice presidents, the vice-chancellor and the registrar of VIT University and Director & Deputy Director of CNBT for their support in research and providing the necessary facilities.


None declared


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Journal of Critical Reviews
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Niyas Ahamed
Centre for Nanobiotechnology, VIT University, Vellore 632014, Tamil Nadu, India


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