Laboratory Experiment

pubs.acs.org/jchemeduc

Introducing Environmental and Sustainable Chemistry Topics Using

a Nanotechnology Approach: Removing Hazardous Metal Ions by
Means of Humic-Acid-Modified Superparamagnetic Nanoparticles
Delmárcio Gomes da Silva,† Fernando Menegatti de Melo,† Alceu Totti Silveira, Jr.,†
Bruno Constancio da Cruz,‡ Caio Cesar Pestana Prado,‡ Luana Cristina Pereira de Vasconcelos,‡
Vitor Amaral Sanches Lucas,‡ and Henrique Eisi Toma*,†
†
Universidade de São Paulo, Departamento Química Fundamental, Instituto de Química, São Paulo, São Paulo 05508-000, Brazil
‡
Centro Estadual de Educaçaõ Tecnológica Paula Souza, Etec Osasco II, Osasco, São Paulo 06296-220, Brazil
*
S Supporting Information

ABSTRACT: A laboratory experiment has been developed to illustrate environmental

and sustainability aspects, focusing on the wastewater treatment by means of
superparamagnetic nanoparticles functionalized with humic acid. The experiment,
conducted by a group of high school students, involves nanoparticle synthesis and
minor characterization, followed by their interaction with typical metal ion contaminants
in water. Coordination chemistry concepts were explored to help students understand the
experiment, using competitive agents, such as ammonia, for recovering and recycling the
nanoparticles, highlighting the great affinity of humic acid for the metal ions.

KEYWORDS: First-Year Undergraduate/General, High School/Introductory Chemistry, Environmental Chemistry,

Interdisciplinary/Multidisciplinary, Laboratory Instruction, Hands-On Learning/Manipulatives, Inquiry-Based/Discovery Learning,
Colloids, Magnetic Properties, Nanotechnology

■ INTRODUCTION
Nowadays, nanotechnology, environment, and sustainability
Hg(II), Pb(II), Cr(III), Cr(VI), Ni(II), Co(II), Cu(II), Cd(II),
Ag(I), and As(III,V) species.
issues are becoming quite relevant in chemistry courses. In order to perform environmental detoxification, techniques
However, there is a limited number of experiments available involving adsorption, precipitation, ion exchange, reverse
for exploring such aspects in an interactive way. Such challenge osmosis, electrochemical treatments, membrane filtration,
can be rather gratifying for teaching nanotechnology concepts, evaporation, flotation, oxidation, and biosorption processes
exploring their possible application in environmental problems are being used.6 With the advent of nanotechnology, such
using natural resources or alternatives.1 Pursuing this task, this techniques can be greatly improved by incorporating versatile
didactic experiment focuses on the nanoparticles chemistry and nanoparticles, nanomaterials and nanosensors in the proc-
explores their association with humic acidsan important ess.7−9
natural substancefor removing hazardous metal ions from In this regard, VanDorn et al.10 have employed magnetite
contaminated water in the environment. nanoparticles (Fe3O4) to adsorb arsenic ions and for removing
The experiment also calls the attention to the water them from the contaminated solution by making use of their
management problem, reinforcing the challenges of developing magnetic properties. Furlan et al.11 have impregnated magnetic
environmentally acceptable technologies and the importance of
nanoparticles with activated carbon, introducing a very
keeping improving educational strategies.2−4 In this context, it
should be noticed that among the several problems of water interesting didactic experiment to illustrate the use of such
contamination, a major concern is brought about by the heavy materials in the purification of water.
metals. They are particularly troublesome because, unlike most Herein, we are exploring a new concept, starting from the
organic species, they are not biodegradable and can accumulate functionalization of Fe3O4 nanoparticles with humic acids, in
in living tissues. Moreover, heavy, hazardous metals are not
only introduced in aquatic systems from the natural wear of Received: March 9, 2016
soils and rocks5 but also result from human activities such as Revised: August 5, 2016
mine processing and the industrial use of substances containing Published: September 23, 2016
© 2016 American Chemical Society and
Division of Chemical Education, Inc. 1929 DOI: 10.1021/acs.jchemed.6b00172
J. Chem. Educ. 2016, 93, 1929−1934
Journal of Chemical Education Laboratory Experiment

order to generate a water-soluble nanomaterial exhibiting high As major components of natural organic matter, humic
capacity to sequester metal ions. substances are involved in chemical interactions in soil and
Magnetite is a rather common raw mineral, however in the aqueous systems. They are widely present in natural waters and
nanometric scale it is better prepared by synthetic routes. Its soils, and allow plants to extract metal elements from their
great advantage is associated with the easy recovery and environments.17 Humic substances are usually divided into two
processing by using a magnetic field or a commercial magnet. fractions: humic acid and fulvic acid. The former is soluble in
However, it should be noticed that magnetite nanoparticles are alkaline solutions but insoluble in acidic solutions, whereas the
prone to air oxidation and aggregation in aqueous media, latter, of a lower molecular weight, is soluble in both alkaline
requiring their surface modification with charged polymers, and acidic solutions. HA has a skeleton of alkyl and aromatic
polyelectrolytes or surfactants.12,13 In this sense, the use of units supporting carboxylic acid, phenolic hydroxyl, and
humic acid (HA) can be particularly rewarding, because it as a quinone functional groups, as shown in Figure 1, and it is an
natural antioxidizing compound exhibiting a great affinity for abundant organic compound in natural aqueous systems.16
metal ions,14,15 as well as for Fe3O4 nanoparticles (MagNP), These functional groups are suitable for binding metal ion
because of its rich phenolic content (Figure 1). The sorption of contaminants from water.18,19
HA on the Fe3O4 nanoparticles (MagNP/HA) also enhances In this work, we describe a 3 h laboratory experiment which
the stability of the nanodispersions, preventing their was successfully applied to high school students, focusing on
aggregation.16 the preparation of magnetic nanoparticles modified with humic
acid, and their application as absorbers for metal ion
contaminants in water. Before the laboratory class, the students
attended a 2 h lecture/discussion on environmental chemistry,
sustainability and nanotechnology issues. As illustrated in
Figure 2, the students could see that humic acid has a natural
potential to capture ions in a plant root acting as a natural soil
conditioner. Exploring this concept, the ability of HA to form
complexes with metal ions was used in a synergic association
with magnetic nanoparticles for the treatment of water
containing some most common contaminants, such as Ni2+,
Co 2+, Cu 2+ , Hg 2+, and Pb 2+ ions. Operational issues,
encompassing the synthesis of MagNP and its functionalization
with HA were also dealt with in this preliminary lecture.
During the laboratory activity, the students performed the
synthesis of nanoparticles and they also conducted remediation
Figure 1. Structure of humic acid showing the aromatic units with
tests on contaminated water samples provided by the
carboxylic acid, phenolic hydroxyl, and quinone functional groups.
instructor. After the experiment, the students discussed the
results and a brief scientific report was requested, including

Figure 2. Capturing sustainable chemistry concepts by observing a natural phenomenon and developing new ideas with a mimicking nanomaterial
for removing heavy metals from water.

1930 DOI: 10.1021/acs.jchemed.6b00172

J. Chem. Educ. 2016, 93, 1929−1934
Journal of Chemical Education Laboratory Experiment

Figure 3. (A) Students preparing the reagents for the synthesis of MagNP/HA. (B), (C) Students conducting the process of purification to remove
the excess of humic acid that is not bound to nanoparticles.

their own evaluation of the subject and the pedagogical strategy (brown color washings, as shown in Figure 3) and then dried in
employed. a vacuum oven. Typically, 5.0 g of MagNP/HA were dispersed

■ EQUIPMENT AND REAGENTS

In the laboratory work, the following facilities have been
in 100 mL of water, in order to obtain a 5.0% (w/w)
suspension.
Preparation of Heavy Metal Solution
employed: a semianalytical balance, a hot plate with mechanical The students received 50 mL of a previously prepared heavy
or magnetic stirrer, a three-neck round-bottom flask equipped metal (chlorides or sulfates) solution containing Ni2+ (50
with a reflux condenser, 0−100 °C thermometer, and ppm), Co2+ (50 ppm), Cu2+ (50 ppm), Pb2+ (35 ppm), and
conventional laboratory glassware. Hg2+ (5 ppm).
Depending on the existing support, the experiment can be Adsorption of Heavy MetalsWater Purification
further extended in order to include specific measurements for
the characterization of the nanoparticles, such as DLS (dynamic As shown in Table 1, 1.0 mL samples of the metal containing
light scattering) for size evaluation and EDS (energy dispersive solution were transferred into four test tubes. Each student was
spectra) for metal ion analysis. Standard colorimetric methods responsible for a test tube. After adding different volumes of
can also be employed for this purpose as an alternative method MagNP/HA (5.0% w/w), the pH was adjusted to 9 using
for qualitative determination.20 NH4OH (25%), and the final volume of the mixture was

■
adjusted to 5.0 mL. The use of the ammonia solution provided
EXPERIMENTAL PROCEDURE a complexing media capable of preventing the precipitation of

Synthesis of MagNP/HA Table 1. Sample Components for the Evaluation of the Metal
The synthesis of MagNP/HA was based on a modified Removal Efficiency Using Variable Amounts of MagNP/HA
chemical coprecipitation method,13 starting from 1.5 g of
humic acid (Sigma-Aldrich), dissolved in 300 mL of water and Sample Tubes
25 mL of NH4OH (25%). This mixture was heated to 90 °C in Parameters 1 2 3 4 Control
a 500 mL three-neck round-bottom flask equipped with a reflux Solution Heavy 1.0 1.0 1.0 1.0 1.0
condenser (solution A). In the other flask, 4.28 g of FeSO4· Metals, mL
7H2O and 8.56 g of FeCl3·6H2O were previously dissolved in MagNP/HA 1.0 0.5 0.25 0.1
suspension,
100 mL of water (solution B). This solution was rapidly added mL
to solution A. The mixture was mechanically (600 rpm) stirred NH4OHconc, 0.15 0.15 0.15 0.15
during the process and the temperature was maintained at 90 ± mL
5 °C for 30 min and then cooled to room temperature. The pH, Adjusted 9−10 9−10 9−10 9−10
black MagNP/HA precipitate was confined at the bottom, Water, mL 2.85 3.35 3.60 3.75 4.0
using an external magnet. The wet paste was rinsed several Final Volume, 5.0 5.0 5.0 5.0 5.0
times with water in order to remove the excess of humic acid mL

1931 DOI: 10.1021/acs.jchemed.6b00172

J. Chem. Educ. 2016, 93, 1929−1934
Journal of Chemical Education Laboratory Experiment

Figure 4. Left: Flasks containing different amounts of nanoparticles MagNP/HA (% w/w). Right: After the magnetic separation.

the metal hydroxides, by forming amine complexes with the The zeta potential of the MagNP/HA measured at pH 9 was
metal ions. It should be noticed that humic acid is a strong −52.1 mV, indicating that the magnetic nanoparticles were
complexing agent, and even under diluted conditions, it is able
to displace the NH3 ligands from the metal ion coordination negatively charged, as shown in Figure 5A. Such a large zeta
sphere. potential is important to prevent the coalescence of the
The mixture was stirred for 30 min and the magnetic nanoparticles, facilitating the sorption of the positively charged
nanoparticles (MagNP/HA) containing the sequestered metal
ions were separated from the mixture using a neodymium metal ions. The size distribution of MagNP/HA was probed by
(Nd2Fe14B) permanent magnet. Then each student separated
an aliquot of this supernatant (Figure 4) and delivered to the
instructor, in order to perform the analysis of the residual
concentration of the metals by EDS (X-ray fluorescence). As a
control, the instructor also prepared standard samples, by
transferring 1.0 mL of solution containing the metals into
another test tube and adjusting the final volume to 5.0 mL with
water.

■ HAZARDS
Heavy metals are toxic substances. The instructor should
prepare the hazardous metals solution beforehand in order to
reduce the student exposure, with the special care required for
handling mercury and lead salts. Standard procedures for safe
handling of chemicals should be followed. Students must wear
lab coats, nitrile gloves, and goggles at all times. The
neodymium magnets used for iron removal are very strong
and should be handled with care to avoid pinching.

■ RESULTS AND DISCUSSION

Characterization of MagNP/HA
For the purposes of this experiment, it is not necessary to
characterize the MagNP/HA nanoparticles. For this reason,
these results were presented as a Supporting Information for
the students, to be discussed in the proper occasion. The
characterization was performed by the team of supervisors of
this project, but every procedure and result was discussed with
the students along the laboratory experiments. In particular, the
evaluation of the zeta potential was highly rewarding, allowing Figure 5. (A) Zeta potential found (−52.1 mV) indicates that the
to understand the role of HA in the suspension stability and the magnetic nanoparticles are negatively charged. (B) Size distribution of
changes resulting from the binding of the metal ions. MagNP/HA reveals an average diameter of 72.5 nm.

1932 DOI: 10.1021/acs.jchemed.6b00172

J. Chem. Educ. 2016, 93, 1929−1934
Journal of Chemical Education Laboratory Experiment

Figure 6. Metal ions removal observed for the several samples (T1−T4) in relation to the control experiment (IC).

dynamic light scattering (DLS) as shown in Figure 5B revealing particles are present in small amounts, the capture of Hg2+ is
a maximum distribution rate at 72.5 nm (22.5%). not efficient because of the presence of ammonia in excess,
Removal of ContaminantsApplication in Environmental generating [Hg(NH3)2]2+ species. The same observation
Remediation applies to Cu2+ ions. For this reason, the reaction involved
would be better expressed as
After each student received the results, the residual
concentrations of metal ions in the supernatant were organized M2 +(aq) + n NH3 ⇌ [M(NH3)n ]2 +
in a table and graphics and discussed at the end of the
laboratory session. Typical results from the analysis of the
supernatant after treatment with MagNP/HA by EDS can be MagNP MagNP
seen in Figure 6. + [M(NH3)n ]2 + ⇌ + n NH3
HA M(HA)x
The results indicate a substantial removal of the metal ions
from all the samples; however, the correct interpretation
requires the understanding of the chemical equilibrium Recycling the Particles
involved. The quantitative data collected in Table 2 allows to Recycling experiments can be carried out by confining
discuss in more detail the metal ion removal efficiency by magnetically the nanoparticles at the bottom of the test tube,
MagNP/HA. and treating the residue with one drop of concentrated
ammonia solution. After completing the volume with water,
Table 2. Average Analytical Data Obtained by the Students the metal ions released in the process can be easily
for the Metal Removal Rates demonstrated using classical spot tests, for example, with
dithiooxamide and sodium rhodizonate in a qualitative way,20
Average Removal of Metal, %
or by means of the EDS technique. Colorimetric analysis can
Sample Tube Ni Co Pb Cu Hg also be performed;20 however, in this case, we recommend
1 95 91 98 94 62 dealing with just one element, separately, in order to overcome
2 93 88 97 81 44 the interference problems.
3 86 91 95 52 12 It should be noted that the magnetic procedure for confining
4 80 91 92 26 0 and recycling the nanoparticles is very simple and quite
convenient from the point of view of green chemistry and
Although the removal efficiency was nearly comparable for sustainability.21,22 In the experiment, the students were also
Ni2+, Co2+, Cu2+, and Pb2+ ions in Tubes 1 and 2, the results stimulated to think about the fate of the heavy metal ions in
indicated a substantial decrease in the case of the Hg2+ ions. natural water resources and on the essential role of the humic
This observation might look rather puzzling because Hg2+ ions acids as sequestering agents for metal ions in the environment.
are known to form very strong complexes with most common As a final remark, the group of students were invited to
ligands. Actually, this point requires the understanding of soft− participate in a schoolroom seminar program after preparing a
hard acid−base concepts because mercury ions are typically short monograph on sustainable technologies for the treatment
soft, and the dominant complexing groups in humic acids are of water resources. This activity reinforced the very positive
hard or of intermediate nature. In this way, when MagNP/HA evaluation of their learning from this particular experiment.
1933 DOI: 10.1021/acs.jchemed.6b00172
J. Chem. Educ. 2016, 93, 1929−1934
Journal of Chemical Education

■
Laboratory Experiment

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Andreescu, S. Adsorption of Arsenic by Iron Oxide Nanoparticles: A
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Sustainability was also valorized in the experiment, from the (12) Maity, D.; Agrawal, D. Synthesis of Iron Oxide Nanoparticles
choice of recyclable, environmentally correct, humic acid and under Oxidizing Environment and Their Stabilization in Aqueous and
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(13) da Silva, D. G.; Toma, S. H.; de Melo, F. M.; Carvalho, L. V. C.;
heavy metal contaminated water.

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Magalhães, A.; Sabadini, E.; dos Santos, A. D.; Araki, K.; Toma, H. E.
Direct Synthesis of Magnetite Nanoparticles from Iron (II)
ASSOCIATED CONTENT Carboxymethylcellulose and Their Performance as NMR Contrast
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(16) Sparks, D. L. Environmental Soil Chemistry; Academic Press:
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(DOCX) Humic Substances in Soil, Sediment, and Water: Geochemistry, Isolation

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and Characterization; John Wiley & Sons: New York, 1985.
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Metal−Humic Acid Complexes and Its Role in Environmental
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ACKNOWLEDGMENTS (21) Toma, H. E. Developing Nanotechnological Strategies for
Green Industrial Processes. Pure Appl. Chem. 2013, 85 (8), 1655−
The authors would like to thank all the teachers and supervisors 1669.
of Centro Estadual de Educaçaõ Tecnológica Paula Souza and (22) Toma, H. E. Magnetic Nanohydrometallurgy: A Nano-
ETEC OSASCO II, and FAPESP Grant 2013/24725-4 for technological Approach to Elemental Sustainability. Green Chem.
support. 2015, 17, 2027−2041.

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1934 DOI: 10.1021/acs.jchemed.6b00172

J. Chem. Educ. 2016, 93, 1929−1934