Advanced Powder Technology 21 (2010) 461467

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Advanced Powder Technology

journal homepage: www.elsevier.com/locate/apt

Original Research Paper

Preparation of Fe3O4chitosan nanoparticles used for hyperthermia

Jingmiao Qu a, Guang Liu a, Yiming Wang a, Ruoyu Hong a,b,*
a
b

College of Chemistry, Chemical Engineering & Materials Science, and Key Laboratory of Organic Synthesis of Jiangsu Province, Soochow University, SIP, Suzhou 215123, China
Key Lab of Multiphase Reaction, Inst. of Process Eng., Chinese Academy of Sciences, Beijing 100080, China

a r t i c l e

i n f o

Article history:
Received 31 October 2009
Received in revised form 14 January 2010
Accepted 26 January 2010

Keywords:
Fe3O4
Chitosan
Nanoparticle
Hyperthermia

a b s t r a c t
The Fe3O4chitosan nanoparticles with core-shell structure have been prepared by crosslinking method.
Oleic acid modied Fe3O4 nanoparticles were rstly prepared by co-precipitation then chitosan was
added to coat on the surface of the Fe3O4 nanoparticles by physical absorption. The Fe3O4chitosan nanoparticles were obtained by crosslinking the amino groups on the chitosan using glutaraldehyde. Transmission electron microscopy showed that the Fe3O4chitosan nanoparticles were quasi-spherical with
a mean diameter of 10.5 nm. X-ray diffraction pattern and X-ray photoelectron spectra indicated that
the magnetic nanoparticles were pure Fe3O4 with a cubic inverse spinel structure. The modication using
chitosan did not result in a phase change. The binding of chitosan to the Fe3O4 nanoparticles was also
demonstrated by the measurement of fourier transform infrared spectra and thermogravimetric analysis.
Magnetic measurement revealed that the saturation magnetization of the composite nanoparticles was
30.7 emu/g and the nanoparticles were superparamagnetic at room temperature. Furthermore, the
inductive heating property of the composite nanoparticles in an alternating current magnetic eld was
investigated and the results indicated that the heating effect was signicant. The Fe3O4chitosan nanoparticles prepared have great potential in hyperthermia.
2010 The Society of Powder Technology Japan. Published by Elsevier B.V. and The Society of Powder
Technology Japan. All rights reserved.

1. Introduction
Nanoparticles (NPs) have been a topic of intense research mainly
because of their unique physical and chemical properties compared
with their bulk counterparts. The case of magnetic NPs is especially
interesting as the NP size is comparable to the size of a magnetic domain, which would result in unusual magnetic phenomena known
as superparamagnetism [1]. Magnetite (Fe3O4) is an important kind
of magnetic material having cubic inverse spinel structure which
has been attracting increasing attention because of its wide use in
magnetic recording [2], ferrouid [3], catalyst [4] and biomedical
applications, such as magnetic resonance imaging (MRI) [57],
bio-separation [8,9], drug targeting [1012] and hyperthermia
[1315].
For biomedical applications Fe3O4 NPs are often treated with surface modication. On the one hand the modication could increase
the chemical stability of the Fe3O4 NPs; on the other hand it could
improve the biocompatibility of the NPs. A variety of materials have
been reported to modify Fe3O4 NPs, such as precious metals [16,17],

* Corresponding author. Address: College of Chemistry, Chemical Engineering &

Materials Science, and Key Laboratory of Organic Synthesis of Jiangsu Province,
Soochow University, SIP, Suzhou 215123, China. Tel.: +86 512 77000797; fax: +86
512 6588 0089.
E-mail address: hongruoy@126.com (R. Hong).

silica [18], carbon [19] and biopolymers [2023]. Chitosan is a partially acetylated glucosamine biopolymer with many useful features such as hydrophilicity, biocompatibility, biodegradability
[24,25]. In addition, the amino groups on the chitosan can also be
used for further functionalization with specic components, such
as various drugs, specic binding sites, or other functional groups.
Thus, it is a suitable kind of polymer to be used to modify the
Fe3O4 NPs. Many investigations on chitosan modied magnetic
NPs used for biomedical applications have been reported. Liang
and Zhang [26] prepared superparamagnetic Fe3O4 NPs decorated
with carboxymethylated chitosan which were used for covalent
conjugation of papain. Feng et al. [27] synthesized monodisperse
chitosan/polyacrylic acid/Fe3O4 NPs which could be used for MRI.
Donadel et al. [28] prepared iron oxide magnetic particles coated
with chitosan used for hyperthermia. However, the saturation magnetization (22 emu/g) of the composite NPs is low and the heating
effect is not desirable. Zhao et al. [29] prepared chitosanFe3O4
NPs in inverse emulsion and investigated their inductive heating
property in AC magnetic eld. The problem about this method is
that the use of organic solvent is not environment friendly and the
dispersity of the chitosanFe3O4 NPs prepared is poor.
For biomedical applications such as hyperthermia, it is required
that the Fe3O4 NPs have high saturation magnetization, uniform
particle size and superparamagnetism. In the investigation, Fe3O4
NPs were prepared using co-precipitation method and oleic acid

0921-8831/$ - see front matter 2010 The Society of Powder Technology Japan. Published by Elsevier B.V. and The Society of Powder Technology Japan. All rights reserved.
doi:10.1016/j.apt.2010.01.008

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J. Qu et al. / Advanced Powder Technology 21 (2010) 461467

was used to prevent the agglomeration among the NPs. For preparing the chitosanFe3O4 NPs, the chitosan rst coated on the surface
of the NPs through physical absorption then glutaraldehyde was
used to crosslink the chitosan and the Fe3O4chitosan NPs were
obtained. The as-prepared composite NPs had good crystal structure, small particle size and high saturation magnetization. A schematic illustration for the formation of the Fe3O4chitosan NPs is
shown in Fig. 1. The inductive heating property of the composite
NPs in an alternating current (AC) magnetic eld was investigated
and the related factors were discussed.

2. Experiments
2.1. Materials
Iron(III) chloride hexahydrate (FeCl3
6H2O), iron (II) sulfate heptahydrate (FeSO4
7H2O), aqueous ammonia (25%), chitosan (molecular weight: 2  105), acetic acid, ethanol were all purchased from
the SCRC (Sinopharm Chemical Reagent Co., Ltd.). All chemicals are
analytical grade without further purication. Deionized water was
used throughout the experiments.
2.2. Preparation of Fe3O4chitosan NPs
Synthesis of the Fe3O4 NPs was based on our previous study
with some modications. In a typical procedure, a mixture of
FeCl3
6H2O (0.046 mol) and FeSO4
7H2O (0.023 mol) was dissolved
into 150 mL deionized water in 250 mL three-necked ask and Ar
was imported for 3 min to extrude the air. Then 20 mL ammonium
hydroxide (25%) was added quickly into the iron solution under
vigorous stirring. After 30 min 3 mL oleic acid was added into the
mixture to modify the Fe3O4 NPs and the mixture was heated to
75 C. Kept at this temperature for 1 h under Ar protection then
cooled to room temperature. The Fe3O4 NPs were collected through
magnetic separation and washed with deionized water and ethanol
three times, respectively, then dried at vacuum conditions.
For preparation Fe3O4chitosan NPs, chitosan (0.5 g) in 100 mL
of 2.0 wt% acetic acid solution, 1.5 g oleic acid modied Fe3O4 NPs
were added into a three-necked ask. The mixture was treated
with ultrasonic for 10 min and stirred for 20 min. Then 2 mL of glutaraldehyde solution (25 wt%) was added into the mixture at 40 C
and crosslinking reaction was kept for 3 h. After the reaction the
composites were collected through magnetic separation and were
washed by deionized water and ethanol for several times. The
products were dried at vacuum conditions.

Fig. 1. A schematic illustration for the formation of Fe3O4chitosan NPs.

2.3. Characterization
Fourier transform infrared (FT-IR) spectroscopy was recorded
on a Nicolet FT-IR Avatar 360 spectrophotometer to analyze the
interaction between chitosan and Fe3O4 NPs. The transmission
electron micrographs were obtained with a Hitachi H-600-II transmission electron microscope (TEM) to observe the particle size and
morphology of the Fe3O4chitosan NPs. X-ray powder diffraction
(XRD) patterns were obtained on a X Pert-Pro MPD diffractometer
using Cu-Ka radiation (k = 0.15405 nm) to determine the crystal
structure of the composites. The chemical composition of the
Fe3O4chitosan NPs was determined with X-ray photoelectron
spectroscopy (XPS) on an AXIS HSi spectrometer. Thermogravimetric analysis (TGA) was performed on a PerkinElmer TGA-7 thermogravimetric analyzer to determine the magnetic content. The
magnetic properties of the composite were measured on a BHV55 vibrating sample magnetometer (VSM). The heating experiments of the Fe3O4chitosan NPs in AC magnetic eld were performed on a medium frequency induction heating apparatus
(Chien-Wha Co., Ltd, Dongguan, China).
3. Results and discussion
Fig. 2 shows the FT-IR spectrum of pristine Fe3O4 NPs (b), oleic
acid modied Fe3O4 NPs (c) and Fe3O4chitosan NPs (a). The characteristic peaks of Fe3O4 at 580 cm 1 could be observed in all of the
three curves which indicated that the NPs were Fe3O4. The peaks at
around 3400 cm 1 and 1632 cm 1 in curve (b) are due to the adsorbed water in the sample. The characteristic peaks of oleic acid
can be observed at 2921, 2849 in curve (c) which can be assignable
to the symmetric and asymmetric stretch of CH3 and CH2. The
peaks at 1604 cm 1 and 1402 cm 1 corresponded to the asymmetric and symmetric stretching band of COO of oleate [30]. The
absorption peak near 1110 cm 1 can be ascribed to the vibration
of OCO bond of the oleate. The peaks of adsorbed water were
not detected in curve (c) indicating that the sample c was dry enough and no adsorbed water existed. These observations indicated
that the oleic acid has changed into oleate and coated onto the surface of the Fe3O4 NPs. From (a) the characteristic peaks of chitosan
could be observed. The band of 1620 cm 1 was assigned to NH
bending vibration and the peak of 1392 cm 1 to CO stretching
of primary alcoholic group in chitosan [31]. The results suggested
the integration of Fe3O4 NPs and chitosan in the Fe3O4chitosan
NPs.

Fig. 2. The FT-IR spectrum of pristine Fe3O4 NPs (b), oleic acid modied Fe3O4 NPs
(c) and the Fe3O4chitosan. NPs (a).

J. Qu et al. / Advanced Powder Technology 21 (2010) 461467

Fig. 3(1) shows the XRD patterns of pristine Fe3O4 NPs (a), oleic
acid modied Fe3O4 NPs (b) and the Fe3O4chitosan NPs (c). A series of characteristic peaks for Fe3O4 (2h = 30.2, 35.5, 43.1, 53.4,
57.0, and 62.6) were observed in all of the samples. These peaks
are consistent with the standard pattern for Fe3O4 (JCPDS #19629) with a cubic inverse spinel structure. The results indicated
that the modication did not change the crystal structure of the
Fe3O4 NPs. The average crystallite size D is calculated using the DebyeSherrer equation [32], D = Kk/(b cos h), where K is a constant
(K = 0.9 for Cu-Ka), k is wavelength (0.15405 nm for Cu-Ka), b is
the peak width of half-maximum and h is the diffraction angle.
The crystallite size of oleic acid modied Fe3O4 NPs thus obtained
from this formula was about 8.85 nm. In the synthesis of Fe3O4,
maghemite (c-Fe2O3) often formed, showing the similar XRD

463

pattern to Fe3O4. To further conrm the structure of the product,

the Fe3O4chitosan NPs were examined by X-ray photoelectron
spectra (XPS) which is shown in Fig. 3(2). The gure shows that
the binding energies relating to Fe 2p3/2, Fe 2p1/2, are about
711.0, 724.3 eV, respectively. These binding energies data are consistent with the reported values of Fe3O4 in the literature [33].
Fig. 4 shows the TEM images of the Fe3O4chitosan NPs. The
size distribution was determined by measuring diameters of one
hundred NPs randomly selected on the TEM micrographs which
is shown in Fig. 5. It can be seen that the pristine Fe3O4 NPs were
polydisperse and seriously aggregated. After surface modication
by oleic acid the particles maintained their original spherical shape
with a size of about 8.8 nm and the particle size is very uniform.
The particle size was in accordance with the value obtained from

Fig. 3. (1) XRD patterns of pristine Fe3O4 NPs (a), oleic acid modied Fe3O4 NPs (b) and the Fe3O4chitosan NPs (c). (2) XPS spectra of Fe3O4chitosan NPs. The expanded
spectra are for Fe 2p and O 1s.

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J. Qu et al. / Advanced Powder Technology 21 (2010) 461467

Fig. 4. TEM images of pristine Fe3O4 NPs (a), oleic acid modied Fe3O4 NPs (b), the Fe3O4chitosan NPs (c) and (d) is the magnied image of the Fe3O4chitosan NPs.

Fig. 5. Size distribution of the oleic acid modied Fe3O4 NPs (a) and the Fe3O4chitosan NPs (b).

TEM. From (c) one can see that the Fe3O4chitosan NPs were quasispherical and had good dispersity. The average size of the Fe3O4
chitosan NPs was about 10.5 nm which was larger than the oleic
acid modied Fe3O4chitosan NPs. The enlargement of the NPs size
indicated the formation of the Fe3O4chitosan NPs with core-shell
structure and the chitosan shell was about 1.7 nm. From the magnied image a slight aggregation can be observed which was due to
the crosslinking among the different NPs. from the curves it can be
seen that the size distribution of Fe3O4chitosan NPs was narrow.

Magnetic properties of Fe3O4chitosan NPs were characterized

by vibrating sample magnetometer (VSM). The magnetic hysteresis
curves are shown in Fig. 6, (a) pristine Fe3O4 NPs (b) oleic acid
modied Fe3O4 NPs and (c) Fe3O4chitosan NPs. It can be seen that
no coercivity or remanence could be observed for all of the three
samples, suggesting the superparamagnetic properties of the
Fe3O4 NPs. This can be ascribed to the small size of NPs which were
smaller than the superparamagnetic critical size (25 nm) [34]. The
saturation magnetization (Ms) of the pristine Fe3O4 NPs was

J. Qu et al. / Advanced Powder Technology 21 (2010) 461467

465

Fig. 6. Magnetic hysteresis curves for pristine Fe3O4 NPs (a), oleic acid modied
Fe3O4 NPs (b) and the Fe3O4chitosan NPs (c) at room temperature.

74.3 emu/g. The high saturation magnetization indicated the good

crystal structure. The saturation magnetization of the Fe3O4chitosan NPs was 30.7 emu/g which was lower than the pristine Fe3O4.
This was due to the existence of the large amount of diamagnetic

Fig. 7. TGA of Fe3O4chitosan NPs (b) and oleic acid modied Fe3O4 NPs (a).

chitosan in the Fe3O4chitosan NPs [35]. However, the values of

the saturation magnetization were much larger than the result

Fig. 8. Heating curves of the Fe3O4chitosan NPs. (1) Experimental setup for calorimetric measurement. (2) Heating curves for different samples at xed apparent current
(I = 300 A) and concentration (10 mg/mL), (a) neat Fe3O4 NPs (b) Fe3O4chitosan NPs. (3) Heating curves the Fe3O4chitosan NPs with different concentration at xed
apparent current (I = 300 A), 10 mg/mL, 20 mg/mL, 30 mg/mL. (4) Heating curves with different apparent current at xed concentration (C = 10 mg/mL), 300 A, 500 A, 700 A.

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J. Qu et al. / Advanced Powder Technology 21 (2010) 461467

reported in the literature (22 emu/g) [28]. Therefore, this value was
much more acceptable for the application of hyperthermia. The
magnetic content in the Fe3O4chitosan NPs was further determined using TGA (Fig. 7).
TGA curves of the oleic acid modied Fe3O4 NPs and Fe3O4
chitosan NPs are shown in Fig. 7, (a) oleic acid modied Fe3O4
NPs and (b) the Fe3O4chitosan NPs. The samples were measured
in the condition of nitrogen atmosphere at the heating rate of
15 C/min until the temperature elevated to 800 C. The weight
loss rate of the oleic acid modied Fe3O4 NPs was about 15% when
the temperature was 800 C which indicated the content of oleic
acid coated on the Fe3O4 NPs was about 15%. The residue weight
of the Fe3O4chitosan NPs was 44% indicating that the Fe3O4 NPs
was 44% in the composite NPs.
The heating experiments in AC magnetic eld were performed
on a medium frequency induction heating apparatus at room temperature. A kerosene thermometer was used to record the temperature to avoid any electrical or magnetic heating of the
temperature probe. Fig. 8(1) shows the experimental setup for
calorimetric measurement. The frequency of AC magnetic eld applied was 80 kHz which is just in the range for biomedical applications (50100 kHz) [34]. The AC magnetic eld strength was
denoted using apparent current (I) which could be tuned at the
range from 100 A to 900 A. A certain amount of NPs were dispersed
in 10 mL deionized water and treated with ultrasonic to obtain
samples with different concentration (C).
Fig. 8(2) shows heating curves for different samples at xed
apparent current (I = 300 A) and concentration (10 mg/mL), (a)
neat Fe3O4 NPs (b) Fe3O4chitosan NPs. From the curves, it can
be seen that the neat Fe3O4 NPs have better heating effect than
Fe3O4chitosan NPs. The temperature of sample (a) could rise from
25 C to 35 C in 1200 s, while the sample (b) could reach 53 C in
the same time. The decrease of the heating effect could be ascribed
to the decrease of the magnetization of the Fe3O4 NPs as a result of
the coating with chitosan.
Fig. 8(3) shows the relation of temperature versus time of the
Fe3O4chitosan NPs with different concentration at xed apparent
current (I = 300 A), 10 mg/mL, 20 mg/mL, 30 mg/mL. It can be seen
that the temperature of sample rose by 10.1 C, 18.9 C and 26.0 C,
respectively, in 1200 s. The sample could reach higher temperature
in the same time when the concentration of the Fe3O4chitosan
NPs was increased. Because the heat source was the magnetic
NPs, in a given time the more the amount of the magnetic NPs
the more the heat could be generated.
Fig. 8(4) shows the heating effect of the Fe3O4chitosan NPs
(C = 20 mg/mL) with different apparent current (300 A, 500 A,
700 A). As can been seen, with increasing the apparent current
the temperature rose more rapidly and elevated to a higher value.
When the apparent current was 300 A, the sample could reach
43.9 C in 1200 s. When the apparent current of 500 A and 700 A
was applied, the temperature of the samples could reach 51.4 C
and 53.1 C, respectively. The results indicated that apparent current has great inuence on heating effect. The AC magnetic eld
was the resource of the energy. More energy of electromagnetic
waves has been absorbed by Fe3O4chitosan NPs in higher magnetic eld. Thus Fe3O4chitosan NPs could generate more heat
and reach a higher temperature value when a large apparent current was applied. So the heating effect could be controlled by
adjusting the value of the apparent current.

4. Conclusions
In this paper, the Fe3O4chitosan NPs with core-shell structure
were prepared by a facile method. The as-prepared composite NPs
had good crystal structure with a cubic inverse spinel structure.

The particle size of the composite NPs was 10.5 nm and the size
distribution was narrow. The Fe3O4chitosan NPs exhibited high
saturation magnetization (30.7 emu/g) and superparamagnetic
property. The interaction of Fe3O4 NPs and chitosan were conrmed using FR-IR. The Fe3O4 content was about 44% in the
Fe3O4chitosan NPs. The heating effect of the Fe3O4chitosan NPs
in AC magnetic eld was also investigated. The results indicated
that the heating effect of the Fe3O4chitosan NPs could be tuned
by changing the concentration and the apparent current. The
Fe3O4chitosan NPs had remarkable heating effect which has great
potential in hyperthermia.
Acknowledgments
The project was supported by the National Natural Science
Foundation of China (NNSFC, No. 20476065, 20736004), the Key
Lab. of Multiphase Reaction of the Chinese Academy of Science
(No. 2006-5), the State Key Lab of Coal Conversion of CAS
(No. 2006-902), the Key Lab. of Organic Synthesis of Jiangsu Prov.
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