Chinese Journal of Chemical Engineering, 16(3) 451ü455 (2008)

Preparation and Analysis of Fe3O4 Magnetic Nanoparticles Used as

Targeted-drug Carriers*

ZHAO Yuanbi (ვၐΣ), QIU Zumin (௎ᆣ੖)** and HUANG Jiaying (ܻ‫ޤ‬࿧)
School of Environmental Science and Engineering, Nanchang University, Nanchang 330031, China

Abstract Fe3O4 magnetic nanoparticles were prepared by the aqueous co-precipitation of FeCl3·6H2O and
FeC12·4H2O with addition of ammonium hydroxide. The conditions for the preparation of Fe3O4 magnetic nanopar-
ticles were optimized, and Fe3O4 magnetic nanoparticles obtained were characterized systematically by means of
transmission electron microscope (TEM), dynamic laser scattering analyzer (DLS) and X-ray diffraction (XRD).
The results revealed that the magnetic nanoparticles were cubic shaped and dispersive, with narrow size distribution
and average diameter of 11.4 nm. It was found that the homogeneous variation of pH value in the solution via the
control on the dropping rate of aqueous ammonia played a critical role in size distribution. The magnetic response
of the product in the magnetic field was also analyzed and evaluated carefully. A 32.6 mT magnetic field which is
produced by four ferromagnets was found to be sufficient to excite the dipole moments of 0.05 g Fe3O4 powder 2
cm far away from the ferromagnets. In conclusion, the Fe3O4 magnetic nanoparticles with excellent properties were
competent for the magnetic carriers of targeted-drug in future application.
Keywords targeted-drug, Fe3O4, magnetic particle, nanometre

1 INTRODUCTION In this work, Fe3O4 magnetic nanoparticles with

In chemotherapy, the application of anti-cancer
drugs such as adriamycin is limited in clinics due to
their toxicity for both normal cells and cancer cells.
During these years, people are attempting to improve
the distribution of drug in human body to reduce the
toxicity of these anti-cancer drugs. Thus, drug deliv-
ery systems become a technique to increase the den-
sity of drug in target area and reduce the nocuousness
to normal tissue.
Among various drug delivery systems, the mag-
netic targeted-drug delivery system [14] is considered
the most popular and efficient. In this technique, the
drug carrying magnetic materials like Fe3O4 will be
led to the cancer areas by outside magnetic field after
taken orally or injected through vein. Moreover, Fe3O4
as magnetic material is wildly used in human treat-
ment because of its stable quality, high magnetic re-
sponses and easy achievement. After absorption by
human body, the remanent magnetic particles can be
safely excreted through skin, bile, kidney, etc. So the
most important to the targeted treatment is to obtain
extremely small and uniform Fe3O4 magnetic
nanoparticles with high magnetic response.
Nowadays, many methods have been developed
to prepare Fe3O4 magnetic nanoparticles such as
chemical co-precipitation [58], hydrolyzation [9, 10],
micro-emulsion [1113], etc. Among these, the chemi-
cal co-precipitation may be the most promising one
because of its simplicity and productivity. The reac-
tion principle is simply as: Fe2++2Fe3++8OH ė
ˉ
narrow size distribution were obtained by controlling
the titrating rate of ammonium hydroxide. The diame-
ters of the Fe3O4 magnetic nanoparticles mainly
ranged from 5 to 20 nm. The magnetic response of the
product in the magnetic field was also discussed.
2 EXPERIMENTAL
2.1 Equipment and reagent
2.1.1 Main reagent
Ferrous chloride tetrahydrate (FeCl2·4H2O, AR);
ferric chloride hexahydrate (FeCl3·6H2O, AR); am-
monium hydroxide [AR, 25% (by mass) NH3 in wa-
ter]; polyethylene glycol 4000 (PEG4000, CP);
Span-80, Tween-80 (Sheng Yu Medical Chemistry
Limited Corporation, Shanghai); nitrogen of 99.5ˁ;
ferromagnet (7.5 mm2×9.0 mm,Guangfa Magnetic
Material Corporation, Cixi); deionized water.
2.1.2 Main equipment
FA2104 electric analytical balance, X-ray powder
diffractometer (XRD), dynamic laser scattering ana-
lyzer (DLS, Malvern Zetasizer Nano S90), scanning
electron microscope (SEM, Hitachi X-650), transmis-
sion electron microscope (TEM, Hitachi H-600-II),
SJZP-1 Gauss/Tesla-Meter (ShiBo Co., Shanghai).
2.2 Preparation of Fe3O4 magnetic nanoparticles
The Fe3O4 magnetic nanoparticles were prepared

Fe3O4+4H2O. However, to get high magnetic response
and uniform-sized Fe3O4 magnetic particles is hard,
especially the particle size should be in the nano-size
range. Furthermore, there is litter special research on
preparation of Fe3O4 magnetic nanoparticles used for
targeted-drug [1416].
based on the co-precipitation of Fe3+ and Fe2+ with a
molar ratio of 3Ή2 [17] under the aqueous ammonia
(0.3 molǜL 1) as precipitating agent. Briefly, mixed
ˉ
and dissolved ferrous chloride tetrahydrate and ferric
chloride in 100 ml deionized water were under vigor-
ous stiring and with the protection of nitrogen to

Received 2007-09-10, accepted 2008-04-30.

* Supported by the Technology Project of Jiangxi Provincial Education Department and Jiangxi Provincial Science Department.
** To whom correspondence should be addressed. E-mail: mzqiu@ncu.edu.cn
452 Chin. J. Chem. Eng., Vol. 16, No. 3, June 2008
prepare total concentration of 0.015 molǜL 1 ferrite
ˉ
solution containing 1g PEG4000 emulsifier. Aqueous
ammonia (0.3 molǜL 1) was dropped slowly into the
ˉ
mixture solution until the pH value was titrated to 11.0
[7, 8, 14]. It can be observed that the solution became
black due to the formation of Fe3O4 particles. The sam-
ple was kept reacting in water bath at 50°C for 30 min
under vigorous stirring and dispersed in ultra-sonicator
for another 30 min. The black mixture was then aged
at 80°C for 1 h. After that, the product prepared was
transferred to a large beaker and dissociated from liq-
uor with the ferromagnet. Finally, the Fe3O4 magnetic
nanoparticles were washed with distilled water until
the pH value descended to 7.0, and were collected in
vacuum drying chamber after being dried for 4 h at
60°C. The above is the standard experimental condi-
tion used through this work, if it was not specified
otherwise.
2.3 Characterization of Fe3O4 magnetic nanopar-
ticles
2.3.1 Size and shape
The size and shape of Fe3O4 magnetic nanoparticles
was observed directly by TEM. The electron acceleration
voltage was 60 kV. The sample was prepared as follows:
a few prepared Fe3O4 magnetic nanoparticles were put
in absolute alcohol and decentralized in ultrasonator
for 20 min. It was dropped to carbon-coated copper
grid and dried at room temperature. The electron mi-
croscope sample was observed under the TEM.
2.3.2 Structure of crystal
A crystallographic study of Fe3O4 magnetic
nanoparticles was performed on X-ray powder dif-
fractometer using Cu (KĮ, Ȝ˙0.15406 nm) target,
graphite monochromator. Working voltage was 40 kV
and working electrical current was 30 mA with scan
speed of 8°C ǜmin 1.
ˉ
2.3.3 Particle size distribution
The volume-average diameter and size distribution
of Fe3O4 magnetic nanoparticles were measured by
DLS. Before measurement, the prepared magnetic poly-
meric latexes were highly diluted and filtered through
a 0.45 mm filter to remove impurities, then the sample
was charged into a thermostated scattering cell at 25°C.
2.3.4 Magnetic density
The magnetic density of ferromagnet was meas-
ured by SJZP-1 Gauss/Tesla-Meters.
2.3.5 Magnetic response
The relative magnetic response of Fe3O4 magnetic
nanoparticles was measured using an electronic balance
[18]. 0.05 g of magnetic powder was placed in the bal-
ance 2 cm below a magnet of known strength, and the
reading on the balance in the unit of g is used as the
index of the magnetic response of the sample powder.
3 RESULTS AND DISCUSSION
3.1 Effect of alkali
The alkali as precipitator plays an important role
in the reaction system. In this work, NaOH and
NH4OH were compared and evaluated. pH value sal-
tated and jumped intensely as NaOH was being added,
so the final pH value was uncontrollable; When
NH4OH was used, the terminal pH value could be
controlled exactly due to the smooth increase of the
pH value. Therefore, NH4OH was more stable and
controllable than NaOH. Table 1 shows the effect of
the alkali on the particle diameter and magnetic re-
sponse. Compared to NaOH, the particle average di-
ameter is obviously smaller with NH4OH, because
ˉ
NaOH has a higher concentration of OH and a
higher pH value with the same concentration of alkali,
which will accelerate the reaction and lead to large
ˉ
particle size. On the contrary, NH4OH releases OH
gradually to make the particle size controllable. As the
stability and the particle size are involved, NH4OH is
selected as alkali source, though the magnetic re-
sponse of magnetic nanoparticles is not as excellent as
that using NaOH.
Table1 Effect of alkali on preparation
Alkali
Average diameter Magnetic response
ķ
Colour of
/nm /g product
NaOH 24.3 0.1120 black
NH4OH 11.6 0.0837 black
ķ Magnetic response is out of 0.05 g magnetic powder with
the ferromagnets 2 cm away.
3.2 Effect of emulsifier
The difficulty in preparing Fe3O4 magnetic
nanoparticles by chemical co-precipitation is the ten-
dency of agglomeration of particles because of the
extremely small particle size leading to great specific
surface area and high surface energy. Appropriate
emulsifier should be employed to make particles sta-
ble and dispersive in the solution. In this study, three
kinds of emulsifier (Span-80, Tween-80, PEG4000)
were analysed.
As shown in Table 2, PEG4000 presents the best
performance, Tween-80 second and Span-80 last.
Table 2 Effect of emulsifier
Emulsifier Result and phenomenon
Span-80Difficult to separate from liquor under the effect of
ferromagnet, the magnetic particles conglutinated with
Span-80 emulsifier to form agglomeration and made the
solution turbid.
Tween-80 The phenomenon of separation was more obvious than
in Span-80, and most of the product was magnetized to
the bottom, but the upper solution presented brown due
to some floccule suspension.
PEG4000 Isolated from liquor completely under the effect of
ferromagnet, the magnetic particles were all magnetized
to the bottom, and the upper liquor was limpid and clear.
3.3 Effect of dropping rate of aqueous ammonia
In this study, the dropping process of aqueous
ammonia was evaluated as an important part. Initially,
 Chin. J. Chem. Eng., Vol. 16, No. 3, June 2008 453

aqueous ammonia was added quickly (about 60 drop Table 3 Effect of concentration of Fe3+ and Fe2+
per minute) while a lot of brown depositions appeared Concentration of Average Magnetic
in the solution. When the pH value reached 6.0, the Colour
Fe2+, Fe3+/molǜL 1 diameter/nm response /g
ˉ ķ

colour of deposition changed to black; the titrating 0.010 10.4 0.0803 black brown
should be slowed down and be controlled at a definite
0.015 11.6 0.0837 black
rate until the pH value increased to around 11.0. The
other preparation conditions were the same as the 0.020 12.8 0.0828 black
standard conditions. 0.025 14.3 0.0819 black
Figure 1 shows the variation of the size distribu- 0.030 17.7 0.0833 black
tion under different dropping rates of aqueous ammo- ķ Magnetic response is out of 0.05 g magnetic powder with
nia when the pH value was titrated from 6.0 to 11.0. the ferromagnets 2 cm away.
The average diameter of samples varies from 11.6 to
28.8 nm as the increase of dropping rate. In addition,
the size distribution widens severely. Under the stable is high enough to create a large amount of crystal nu-
and slow dropping rate, the variation in pH is homo- cleus, but the growth of crystal nucleus is considera-
geneous and uniform all over the solution. Thus, the bly slow. Along with the increase of concentration, the
size distribution of Fe3O4 magnetic nanoparticles particle size increases gradually. It is also found that
could be significantly narrowed down, and the uni- the magnetic response doesn’t change much with the
formity in the particles size can be greatly enhanced as total iron concentration increases. So the concentration
of Fe2+ and Fe3+ should be fixed at 0.015 molǜL 1.
ˉ
in Fig. 1 (a). In order to get Fe3O4 magnetic nanopar-
ticles with narrow size distribution, 68 dropsǜmin 1
ˉ

is chosen during the pH titration from 6.0 to 11.0. 3.5 Effect of reaction temperature

Reaction temperatures at 30°C, 40°C, 50°C,

60°C, and 70°C were tested, while other preparation
conditions were the same as the standard conditions.
Table 4 illustrates that the particle size increases with
temperature rising, but not obviously below 50°C. The
maximal magnetic response is obtained at 50°C. The
low magnetic response is resulted from the dimen-
sional effect of quantum [19, 20]. Thus, 50°C is se-
lected as a proper reaction temperature when both par-
ˉ1
(a) 68 dropsǜmin (average diameter˙11.6 nm) ticle size and magnetic response are considered.

Table 4 Effect of reaction temperature

Reaction Average diameter Magnetic response
ķ

Colour
temperature/°C /nm /g
30 10.7 0.0770 black brown
40 11.1 0.0811 black
50 11.6 0.0837 black
(b) 1820 dropsǜminˉ1 (average diameter˙18.0 nm) 60 14.7 0.0764 black brown
70 23.2 0.0628 black brown
ķ Magnetic response is out of 0.05 g magnetic powder with
the ferromagnets 2 cm away.

3.6 Effect of the PEG4000 dosage

The average diameter and magnetic response of

ˉ1
(c) 5560 dropsǜmin (average diameter˙28.8 nm)
Figure 1 Effect of the dropping speed of aqueous ammo-
nia on the size distribution of Fe3O4 nano-particles
3.4 Effect of concentration of Fe3+ and Fe2+
Fe3O4 magnetic nanoparticles were prepared with
different concentrations of Fe2+, Fe3+ in aqueous phase,
and other preparation conditions were the same as the
standard conditions. Table 3 shows that the average
size of Fe3O4 magnetic nanoparticles increases as con-
centration increases. The concentration of 0.010 molǜL 1
ˉ
Fe3O4 magnetic nanoparticles prepared with different
dosages of PEG4000 are summarized in Table 5, and
other preparation conditions are the same as the stan-
dard conditions. It shows that dosage of emulsifier
influences the average diameter very much. As a result,
1 g may be an appropriate dosage when narrow size dis-
tribution and high magnetic response are both wanted.
3.7 The structure and shape of Fe3O4 magnetic
nanoparticles
X-ray diffraction pattern (Fig. 2) of Fe3O4 nanopar-
ticles prepared under standard conditions reveals
454 Chin. J. Chem. Eng., Vol. 16, No. 3, June 2008

Table 5 Effect of the PEG4000 dosage some extent, because the extremely small particle size
Dosage of Average Size Magnetic can get great specific surface area and surface energy.
Colour
PEG4000/g diameter/nm distribution/nm response /g
ķ

0.5 19.8 5120 nm 0.0746 black 3.8 Magnetic response of the Fe3O4 magnetic
1 11.6 520 nm 0.0837 black nanoparticles in the magnetic field
1.5 14.2 540 nm 0.0780 black
While the magnetic response of the magnetic
2 21.8 1060 nm 0.0671 black
material is an important essence for targeted drug in
2.5 38.8 2080 nm 0.0506 black brown clinical application, the magnetic response of the
ķ Magnetic response is out of 0.05 g magnetic powder with product in the magnetic field was analyzed and evalu-
the ferromagnets 2 cm away. ated by an electronic balance. 0.05 g Fe3O4 magnetic
nanoparticles prepared under the standard conditions
were tested. Different from the previous reports in
which the magnetization variation of magnetic parti-
cles were analyzed by vibrating-sample magnetometer
[23], this study evaluated the magnetic response of
magnetic particles through a Gauss/Tesla-Meter to give
more direct data for further research of targeted drug.
Firstly, the distance between ferromagnet and
Fe3O4 powder placed on the balance dish was changed
to explore its influence to magnetic response (Fig. 4).
The maximum density of the ferromagnet itself was
206.5 mT which was tested at 0 cm position from
ferromagnet.
Figure 2 XRD pattern of Fe3O4 nanoparticles

diffraction peaks at (111), (220), (311), (400), (422),

(511), (440), etc. which are the characteristic peaks of
the Fe3O4 crystal with a cubic spinel structure. It is
clear that only the phase of Fe3O4 is detectable. There
is no other phase such as Fe(OH)3 or Fe2O3, which are
the usual products in a chemical co-precipitation pro-
cedure. It could also been learned that particle size is Figure 4 Effect of the distance between ferromagnet and
quite small from the relatively wide half-peak breadth. Fe3O4 powder
Ƶmagnetic density;ƽmagnetic response
With the XRD pattern, the average diameter which
can be evaluated from Scherrer equation [21, 22] (D˙
KȜ/ȕcosș, where K is constant, Ȝ is X-ray wavelength Secondly, the number of the ferromagnets was
and ȕ is the peak width of half-maximum) is obtained changed to investigate the effect on magnetic response
as 11.4 nm. (Fig. 5). All the ferromagnets had the same magnetic
Figure 3 shows TEM photograph of Fe3O4 performance, and the distance between ferromagnets
nanoparticles prepared under standard conditions. It is and Fe3O4 powder was fixed at 2 cm.
worthwhile to note that the size distribution is 814
nm which matches the calculated value by the
Scherrer equation and the result of the DLS very well,
and the nanoparticles are cubic shaped, uniform and
monodispersed. However, there is glomeration of

Figure 5 Effect of the number of the ferromagnets

Ƶmagnetic density;ƽmagnetic response

Figure 4 indicates that the intensity of magnetic

field and the magnetic response of Fe3O4 magnetic
nanoparticles weaken as the distance increases, more-
over, the magnetic response depends on the magnetic
intensity. The magnetic response increases nearly 64
times when the distance is changed from 3 cm to 1 cm.
Thus, in the clinical application, the distance between
Figure 3 TEM image of Fe3O4 nanoparticles ferromagnet and drug should be highlighted especially.
 Chin. J. Chem. Eng., Vol. 16, No. 3, June 2008
Fig. 5 illustrates the magnetic response of Fe3O4
magnetic nanoparticles increases rapidly as the in-
crease of the number of the ferromagets initially, but
not obviously when the number of the ferromagets
exceeds four. Therefore, from the economic point of
view, a 32.6 mT magnetic field which is produced by
four ferromagnets was sufficient to excite the dipole
moments of the 0.05 g Fe3O4 powder 2 cm away from
the ferromagnets.
4 CONCLUSIONS
Fe3O4 magnetic nanoparticles with excellent
properties have been successfully prepared by chemi-
cal co-precipitation technique. From the factors of
average diameter and magnetic response for 100 ml of
reacting phase, the optimum condition for preparation
is fixed as follows: the total concentration of Fe2+,
Fe3+ 0.015 molǜL 1, the molar ratio of Fe3+ and Fe2+
ˉ
3Ή2, PEG4000 emulsifier 1 g, the reaction tempera-
ture 50°C, and the terminal pH value 11.0. Fe3O4
magnetic nanoparticles prepared under the standard
conditions are cubic shaped, dispersive and uniform,
and its average diameter is roughly 11.4 nm with nar-
row size distribution ranged from 520 nm. The 32.6
mT magnetic field was sufficient to excite the dipole
moments of 0.05 g Fe3O4 powder 2 cm away from the
ferromagnets. It is concluded that the obtained Fe3O4
magnetic nanoparticles were qualified as the magnetic
carriers for further tests in targeted-drug therapy.
REFERENCES
1 David, S.S.,“Biomedical applications of nanotechnology-implications
for drug targeting and gene therapy”, Trends Biotechnol., 15 (6),
217224 (1997).
2 Josephson, L., Tung, C.H., Moore, A., Weissleder, R., “High efficiency
intracellular magnetic labeling with novel super-paramagnetic tar-
geted peptide conjugates”, Bioconjugate Chem., 10 (2),186 (1999).
3 Torchilin, V.P., “Drug targeting”, Eur. J. Pharm. Sci., 2 (sup.), 8191
(2000).
4 Xu, H., Song,T., “Development of magnetic drug targeting”, Biomed.
Eng. Foreign Med. Sci., 27 (1), 6164 (2004). (in Chinese)
5 Qiu, X.P., “Preparation and characterization of Fe3O4, magnetic
nanoparticles”, J. Xiamen Uni. (Nat. Sci. Ed.)., 38 (5), 711714
455
(1999). (in Chinese)
6 Tu, G.R., Liu, X.F., Du, G.X., Zhou, X.H., Dang, H.J., “Synthesis
and characteristic determination of nanosized Fe3O4 powder”, Fine
Chem., 21 (9), 641644 (2004). (in Chinese)
7 Zhang,X., Li, X.G., Jiang, B., “Preparation and characterization of
nanometer magnetite”, Chem. Ind. Eng., 23 (1), 4548 (2006). (in
Chinese)
8 Jiang, W.Q., Yang, H.C., Yang, S.Y., Horng, H.E., Hung, J.C., Chen,
Y.C., Hong, C.Y., “Preparation and properties of superparamagnetic
nanoparticles with narrow size distribution and biocompatible”, J.
Magn. Magn. Mater, 283, 210214 (2004).
9 Massart, R.J., “Preparation of magnetite nanoparticles”, IEEE Trans
Magn., 17, 1124711250 (1981).
10 Bee, A., Massart, R., Neveu, S., “Synthesis of very fine magnetite
particles”, J. Magn. Magn. Mater., 149, 69 (1995).
11 Ang, B.C., Iskandar, I.Y., “Synthesis and characterization of mag-
netic iron oxide nanoparticles via w/o microemulsion and Massart’s
procedure”, J. Mater. Process. Technol., 191, 235237 (2007).
12 Zhang, W.J., Zhang, H.F., Li, D.Z., “Preparation of Fe3O4 magnetic
fluid by one-step method with a microemulsion reactor”, Chin. J.
Appl. Chem., 22 (7), 734738 (2005). (in Chinese)
13 Zhou, Z.H., Wang, J., Liu, X., “Synthesis of Fe3O4 nanoparticles
from emulsion”, J. Mater. Chem., 11, 17041709 (2001).
14 Jiang, X.Y., Zhou, C.Sh., Zhang, J.Sh., Yang, J.Y., “Preparation and
analysis of magnetic nanoparticles used as targeted-drug carriers”, J.
Central South Univ. Technol., 34 (5), 516520 (2003). (in Chinese)
15 Du, Y.Q., Zhang, D.S., Ni, H.Y., Gu, N., Yan, S.Y., Tang, Q.S., Jin,
L.Q., Wan, M.L., “Preparation and characterization of Fe3O4
nano-magnetic particles for tumor hyperthermia”, J. Chin. Electron
Microsc, Soc., 24 (6), 608612 (2005). (in Chinese)
16 Goodwin, S., Peterson, C., Hoh, C., Bittner, C., “Targeting and re-
tention of magnetic targeted carriers (MTCs) enhancing intra-arterial
chemotherapy”, J. Magn. Magn. Mater., 194, 132139 (1999).
17 An, Z., Zhu, L., Lin, F., “Synthesis and characterization of ultrafine
Fe3O4 nanoparticles”, J. Harbin Med. Univ., 38 (5), 424425 (2004).
(in Chinese)
18 He, Y.B., Qiu, Z.M., Qiu, J.M., “Study on preparation and property
of nanometer Fe3O4 magnetic fluid”, Chin. J. Process Eng., 6
(suppl.2), 263267 (2006). (in Chinese)
19 Hao, Z., Jing, L., Wang, Z.L., Liu, J.P., Sun, S.H., “Biomagnetic
core/shell FePt/Fe3O4 nanoparticles”, Nano Lett., 4 (1), 187190
(2004).
20 Gruttner, C., Teller, J., Schutt, W., Westphal, F., Schumichen, C.,
Paulke, B.R., Scientific and Clinical Applications of Magnetic Car-
riers, Plenum Press, New York (1997).
21 Warren, B.E., X-ray Diffraction, Dover Publications, New York
(1990).
22 Hong, R.Y., Pan, T.T., Li, H.Z., “Microwave synthesis of magnetic
Fe3O4 nanoparticles used as a precursor of nanocomposites and fer-
rofluids”, J. Magn. Magn. Mater., 303, 6068 (2006).
23 Xu, Z.Z., Wang, C.C., Yang, W.L., Deng, Y.H., Fu, S.K., “Encapsu-
lation of nanosized magnetic iron oxide by polyacrylamide via in-
verse miniemulsion polymerization”, J. Magn. Magn. Mater, 277,
136143 (2004).