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Preparation of Size-Controlled Magnetite Nanoparticles for Hyperthermia of

Cancer

Article  in  Transactions of the Materials Research Society of Japan · January 2009

DOI: 10.14723/tmrsj.34.77

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 Transaction of the Materials Research Society of Japan 　34[1] 77-80 (2009)

Preparation of Size-Controlled Magnetite Nanoparticles for Hyperthermia of Cancer

Zhixia Li, Masakazu Kawashita, Norio Araki*, Michihide Mitsumori** and Masahiro Hiraoka**
Graduate School of Biomedical Engineering, Tohoku University, Sendai 980-8579, Japan
Fax: 81-22-795-3937, e-mail: zhixia@ecei.tohoku.ac.jp
*National Hospital Organization Kyoto Medical Center, Kyoto 612-8555, Japan
**Graduate School of Medicine, Kyoto University, Kyoto 606-8507, Japan

Abstract. Size-controlled magnetite nanoparticles (MNP) with several dozen to several hundred of
nanometers (nm) were prepared for hyperthermia of cancer. Nanoparticles of single-phase
magnetite, as revealed by X-ray diffraction and magnetic property measurement, were synthesized
by mean of an oxidation-precipitation method with iron (II) chloride as starting materials, which
was oxidized to ferric salts by sodium nitrate (NaNO3) in an N2-deaerated sodium hydroxide
(NaOH) aqueous solution. The particles size was controlled by adjusting the concentration of the
reactant and the addition order of NaNO3. The resulting MNP had an average diameter ranging
from 24 nm to 413 nm and their in vitro heat generations were measured in agar phantom under an
alternating magnetic field (100 kHz, 23.9 kAm-1). Temperature rise in the agar phantom for MNP
24 nm in size was ∆T =55 oC in 30 second at a MNP concentration of 58 mg Fe ml-1. For MNP
larger than 24 nm, heat generation decreased as particle size increased. Hysteresis loss mainly
contributed the heat-generating mechanism. It is believed that MNP 24 nm in size is useful for in
situ hyperthermia treatment of cancer.
Key words: Magnetite nanoparticles, Heat generation, Oxidation-precipitation, Hyperthermia, Hysteresis loss

1. INTRODUCTION
Magnetic fluids containing ferromagnetic and/or
ferrimagnetic particles were thought to be useful as
thermo seeds for inducing hyperthermia of cancer
treatment [1, 2]. As well known, the magnetic particles
generate heat under an alternating current (AC) magnetic
field by several physical mechanisms: relaxation loss or
hysteresis loss which strongly depends on the frequency
of the external field as well as the nature of the particles
such as particle size and surface modification [3, 4].
Recent reports have demonstrated that dextran/magnetite
complex (DM) generated enough heat via relaxation loss
when placed in an external AC magnetic field [5]. But it
was difficult to let DM particles stay at the affected area
for a long-term and repeated therapy because of their
smaller particle size (dozens of nanometer). Therefore,
magnetic nanoparticles (MNP) with larger size were
inferred to be useful especially in tumors located deep
inside the body [6].
However, as far as we know, it is hard to find papers
describing the heat-generating properties of a larger MNP
more than 20 nm. Hysteresis loss is thought to contribute
the heating mechanism for the larger MNP under a
clinically available AC magnetic field, but the
relationship between particle size and their hysteresis
loss-derived heating properties is not clear. Therefore, it
is necessary to clarify the relationship between particle
size, strength of magnetic field and heating properties of
nanoparticles as fundamental research. It is expected that
such findings are useful for designing novel thermo seeds
for hyperthermia. In this study, we attempted to prepare
series of MNP larger than 20 nm in aqueous solution and
their heating properties were measured.
2. EXPERIMENTAL
2.1 Preparation of magnetite nanoparticles
Under a given molar ratio of Fe2+/NO3-/OH- =3:1:5,
samples were prepared by the following two procedures.
In procedure 1, a calculated amount of sodium nitrate
(NaNO3) was added into a predetermined volume of
sodium hydroxide (NaOH) aqueous solution (0.1~0.5 M),
which was sufficiently deaerated by N2 gas. And then a
predetermined volume of deaerated iron (II) chloride
(FeCl2) aqueous solution (0.5 M) was added to the
NaNO3-NaOH solution under stirring at a rate of 400 rpm.
After stirring for 15 min, the solution was subjected to
ultrasonic treatment for 10 min and aged at 60 ºC for 5 h.
The precipitates were separated from solution with a
magnet and washed four times with pure water and
ethanol, and then dried at 60 ºC for 24 h. This method
provided samples A and B.
In procedure 2, we changed the addition order of
NaNO3. Briefly speaking, a predetermined volume of
FeCl2 solution (0.1 or 0.5 M) was added slowly into
NaOH solution (0.02 or 0.5 M) under stirring. After
running out of the predetermined volume of FeCl2
solution for 5 min, a calculated amount of NaNO3
solution (0.5 M) was added slowly into the FeCl2-NaOH
solution and further stirred for 10 min, and then aged at
60 oC for 4 or 10 h. This method provided samples C and
D. All the above processes were conducted under N2
atmosphere.
2.2 Characterization
The crystalline phase of the samples was verified by
powder X-ray diffraction (XRD; RINT-2200VL, Rigaku,
Japan) using the following settings: X-ray source,
Ni-filtered CuKα radiation; X-ray power, 40 kV, 40 mA;

77
78 Preparation of Size-Controlled Magnetite Nanoparticles for Hyperthermia of Cancer

scanning rate, 2θ = 2º min-1; and sampling angle, 0.02 º. 3. RESULTS AND DISCUSSION
Particle sizes and crystal morphologies of the samples 3.1 Structure of samples
were observed by transmission electron microscope XRD patterns of all samples prepared are shown in
(TEM; HF2000, Hitachi, Japan) and field-emission Fig.1. The XRD patterns were similar and consisted of
scanning microscope (FE-SEM; S4300, Hitachi, Japan). sharp peaks near 18.32 º, 30.10 º, 35.46 º, 43.08 º, 57.07 º
Particle size distributions for samples A and B were and 62.64 º, which proved that all the synthesized
obtained statistically by measuring the sizes of 100 nanoparticles can be identified as magnetite (Fe3O4) with
nanoparticles from the TEM images, while those of spinel structure [8]. The strong and sharp peaks of sample
samples C and D were obtained from SEM images D indicate that the crystallinity of sample D is high.
because samples C and D were difficult to be observed by These XRD peaks did not shift but became broader in the
TEM owing to their strong agglomeration. The saturation cases of samples A and B, indicating a decreased
magnetization (Ms) and coercive force (Hc) of the crystallite size.
samples were measured by a vibrating sample Fig.2 shows the TEM and SEM photographs of samples.
magnetometer (VSM-5, Toei, Japan) using magnetic Sample A contained nearly spherical particles, sample B
fields up to 800 kAm-1 at room temperature. was composed of square-shaped particles with a little
amount of spherical particles, and samples C and D were
2.3 In vitro heating studies and set up mainly consisted of cubic and octahedral particles. The
The magnetic field used to measure the heating average sizes of the samples A and B were around 24 nm
properties of samples was produced by magnetic and 36 nm, respectively, and approximately display a
core-type alternating magnetic field generator as
described in our previous study [7]. The magnetic field
strength was varied by changing the gap between ferrite
cores from 10 to 250 mm and the detail strength
distribution of magnetic field in gap were also discussed
in the same study [7].
All samples were separately dispersed into hot agar
solution (agar content: 1.1 wt%) in a glass tube followed
by ultrasonic treatment for 2 min, and then solidified in
cold water. The concentration of samples in agar
phantom was fixed at 58 mg Fe ml-1. The tubes with agar
phantom were placed in an alternating magnetic field
(100 kHz, 23.9 kAm-1) and the heat generation of samples
was studied by measuring the temperature rise in agar
phantom with a fluoroptic thermometer (Model-3000,
Luxtron Co., California). Fig.1 XRD patterns of samples.

A B

C D

Fig.2 TEM (A and B) and FE-SEM (C and D) photographs of samples.

 Z. Li et al. 　　　　　　　　　　　　　　　　　　　　　　　　　　　34[1]
Transaction of the Materials Research Society of Japan 77-80 (2009)
uniform morphology. The average sizes of samples C and
D were around 65 nm and 413 nm, respectively. Fig.2
also shows that the agglomeration of particles occurred in
all samples, which is due to the magnetic interaction
between the nanoparticles. The particle size and its
distribution of samples are listed in Table 1. The
percentage ratio of the standard deviation to the mean
size of the particle distribution ranged from 22.5 to 37.3%.
The wide size distribution in sample C might be
responsible for the obvious flocculation of particles
because smaller particles trend to settle in the gap
between larger particles to decrease the surface to volume
ratio as a result of basic tendency of primary particles to
reduce surface energy.
The formation of magnetite follows the
nucleation-growth mechanism [9, 10]: in procedure 1,
because NaNO3 is added into NaOH solution prior to the
addition of FeCl2 solution, a large number of Fe3+ ions are
formed at the initial stage after addition of FeCl2 solution.
When the molecular ratio of Fe2+/Fe3+ becomes 0.5,
which is the characteristic value for the magnetite
structure, the nucleation of magnetite particles easily
occurs and then crystal nuclei of magnetite begin to grow
up to form smaller magnetite nanoparticles. This is
Table I
Size (d (mean) ± ∆d (standard deviation)), size distribution (∆d/d), saturation magnetization (Ms) and coercive force
(Hc) of samples.
Samples Size (nm) ∆d/d (%)
A 24 ± 5.4 22.5
B 36 ± 9.2 25.5
C 65 ± 25.0 37.3
D 413 ± 116.5 28.3
Fig.3 Magnetic hysteresis curves of samples at room temperature.
79
similar to the mechanism for co-precipitation of
magnetite in aqueous solution. In procedure 2, Fe2+ ions
are firstly precipitated to form Fe(OH)2 gel in NaOH
solution, and then the Fe(OH)2 gel transfers to an
intermediate colloid particles called as “Green rust II”
(G-II: iron hydroxides containing Fe2+ and Fe3+ ions) at
pH 6~7 [11]. The iron complexes form easily on the
surface of G-II and subsequently transfer to the crystal
nuclei of magnetite. Then, the G-II is slowly oxidized by
the subsequently addition of NaNO3 solution, and
concentration of Fe3+ ions in the solution increases. When
the ratio of Fe2+/Fe3+ around the surface of G-II is 0.5, the
magnetite nuclei in three axes directions are equivalent
and thus trend to grow up, finally hexahedral and
octahedral nanoparticles of magnetite are formed [8, 11].
The reaction rate, i.e., the generating rate of Fe(OH)2
plays a key role on controlling the size of resulting
magnetite particles. Therefore, compared with procedure
1, MNP larger than 100 nm can be easily obtained in
procedure 2.
3.2 Magnetic properties of magnetite nanoparticles
The hysteresis loops of all samples are shown in Fig.3,
and the saturation magnetization (Ms) and coercive force
Ms (10 -3 Am 2 g -1 ) Hc (kAm-1)
76.5 8.4
79.5 11.1
90.8 15.1
92.0 6.7
80 Preparation of Size-Controlled Magnetite Nanoparticles for Hyperthermia of Cancer
(Hc) of samples derived from Fig.3 are listed in Table I.
Sample A and B showed a Ms near 80 Am 2 kg -1 , while
Ms of samples C and D was higher than 90 Am 2 kg -1
which was near the bulk magnetite, suggesting that
procedure 2 easily give a MNP with higher Ms. Coercive
force Hc was discovered to be strongly size dependent.
The theoretic predicts showed that Hc ∝ d -1 for larger
grain size and Hc ∝ d 6 for an ultrafine particle, the
dividing line (dex) between the two cases was calculated
to be 27 nm [12]. Below the size, Hc will decrease rapidly
as particles size decreases. When Hc of sample is reduced
to be zero, the particles become superparamagnetic in an
external field at a fixed temperature. Above the size of 27
nm, Hc of samples decrease slowly as particles size
increases [12]. Our experimental results were in good
agreement with the theoretic results. Hysteresis loss is
mainly due to the magnetic domain wall motion [13], and
its value is given by the area of the hysteresis loop in an
applied AC field. Therefore, the hysteresis loss tends to
vary depending on the particles sizes that cause the
change in the coercive force Hc.
3.3 In vitro heat generation of magnetite nanoparticles
Fig.4 shows the time-dependent temperature curves of
agar phantoms, in which samples were dispersed, under
AC magnetic field of 100 kHz, 23.9 kAm-1. Temperature
rise in agar phantom containing sample A was ∆T = 55 oC
in the initial 30 second, exhibiting the best heating ability
among four samples. Heat-generating ability decreased
with increasing particle size (i.e., B > C > D). In this
study, the applied external magnetic field strength (23.9
kAm-1) was much higher than the coercive forces of
samples A (8.4 kAm-1), B (11.1 kAm-1), C (15.1 kAm-1)
and D (6.7 kAm-1) and hence the heat generations of these
samples are expected to derive from relaxation loss or
hysteresis loss depending on the particle size. Sample A,
whose average size was nearest to dex, also contains some
smaller superparamagnetic particles (< 20 nm), thus
relaxation loss as well as hysteresis loss become an
important contribution to heat generation. On the other
hand, for samples B, C and D whose average sizes were
larger than dex, hysteresis loss mainly contributes to heat
generation in the AC magnetic field. Consequently,
sample A showed better heat-generating ability than
other three samples in this study.
4. CONCLUSIONS
Magnetite particles with different sizes were prepared
by oxidation-precipitation method according to two
procedures, and their heat generations in an external
alternating magnetic field were investigated. The heat
generation of the magnetite nanoparticles depended
remarkably on the particle size in the applied field. The
heat-generating ability of the particles improved with
decreasing particle size from 413 to 24 nm. The main
contribution arose from the hysteresis loss. The magnetite
particles with average size of 24 nm showed the excellent
heat-generating ability under an alternating magnetic
field of 100 kHz, 23.9 kAm-1, which might be a combined
effect of relaxation loss and hysteresis loss of the
magnetic particles. Therefore, it is believed that MNP
around 24 nm in size is useful for the in situ hyperthermia
treatment of cancer.
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Fig.4 Time-dependent temperature curves of agar
phantoms, in which samples were dispersed, under AC
magnetic field of 100 kHz and 23.9 kAm-1.
Acknowledgements
We thank to Assoc. Prof. Masaaki Doi, Graduate
School of Engineering, Tohoku University, for supplying
the equipment for measurement of magnetic properties of
samples. This work was partially supported by Funds for
Promoting Science and Technology under the Program
for Exploring Advanced Interdisciplinary Frontiers, the
Ministry of Education, Culture, Sports, Science and
Technology, Japan, and Research Grants from the
Kazuchika Okura Memorial Foundation, Japan.
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(Received November 28, 2008 ; Accepted December 20, 2008)