Journal of Colloid and Interface Science 360 (2011) 556–564

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Journal of Colloid and Interface Science

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Highly temperature responsive core–shell magnetic particles: Synthesis,

characterization and colloidal properties
Md Mahbubor Rahman a, Mohamed M. Chehimi b, Hatem Fessi a, Abdelhamid Elaissari a,⇑
a
Université de Lyon, F-69622 Lyon, France, Université Lyon-1, Villeurbanne, LAGEP, UMR 5007, CNRS, CPE, 43 bd 11 Novembre 1918, F-69622 Villeurbanne, France
b
Laboratoire ITODYS (Interfaces, Traitements, Organisation & Dynamiques des Systèmes) Université Paris Diderot, 15 rue Jean-Antoine de Baïf, 75013 Paris, France

a r t i c l e i n f o a b s t r a c t

Article history: Temperature responsive magnetic polymer submicron particles were prepared by two step seed
Received 9 March 2011 emulsion polymerization process. First, magnetic seed polymer particles were obtained by emulsion
Accepted 20 April 2011 polymerization of styrene using potassium persulfate (KPS) as an initiator and divinylbenzne (DVB) as
Available online 27 April 2011
a cross-linker in the presence of oil-in-water magnetic emulsion (organic ferrofluid droplets). Thereafter,
DVB cross-linked magnetic polymer particles were used as seed in the precipitation polymerization of
Keywords: N-isopropylacrylamide (NIPAM) to induce thermosensitive PNIPAM shell onto the hydrophobic polymer
Magnetic emulsion
surface of the cross-linked magnetic polymer particles. To impart cationic functional groups in the ther-
Core-shell morphology
Temperature responsive
mosensitive PNIPAM backbone, the functional monomer aminoethylmethacrylate hydrochloride (AEMH)
Volume phase transition was used to polymerize with NIPAM while N,N0 -methylenebisacrylamide (MBA) and 2, 20 -azobis (2-
pH sensitive methylpropionamidine) dihydrochloride (V-50) were used as a cross-linker and as an initiator respec-
Electrophoretic mobility tively. The effect of seed to monomer (w/w) ratio along with seed nature on the final particle morphology
was investigated. Dynamic light scattering (DLS) results demonstrated particles swelling at below vol-
ume phase transition temperature (VPTT) and deswelling above the VPTT. The perfect core (magnetic)
shell (polymer) structure of the particles prepared was confirmed by Transmission Electron Microscopy
(TEM). The chemical composition of the particles were determined by thermogravimetric analysis (TGA).
The effect of temperature, pH, ionic strength on the colloidal properties such as size and zeta potential of
the micron sized thermo-sensitive magnetic particles were also studied. In addition, a short mechanistic
discussion on the formation of core–shell morphology of magnetic polymer particles has also been
discussed.
Ó 2011 Elsevier Inc. All rights reserved.

1. Introduction
Magnetic particles specially magnetite Fe3O4 and maghemite
c-Fe2O3 encapsulated with functional polymers have attained
much interest due to their potential applications in different areas
such as target drug delivery [1,2], protein purification [3], detec-
tion [4], separation [5], cell labeling [6], biosensor [7], enzyme
immobilization [8] and so forth. Magnetic particles as a solid sup-
port for DNA [9] and RNA [10] separation have also been used in
biotechnology due to the ease of handling, rapid processing and
less chemical requirements.
The encapsulation of magnetic iron oxide particles by polymers
gives particles better qualities that cannot be found in uncoated
magnetic particles. As for example, if the magnetic particles are
coated with functional group containing polymer, polymer chains
⇑ Corresponding author. Fax: +33 (0) 4 72 43 16 82.
E-mail address: elaissari@lagep.univ-lyon1.fr (A. Elaissari).
prefer to stay away from the surface to decrease the intertwines
overlapping. The high osmotic pressure within the functional
groups provides repulsive force when the two particles coated with
polymer come to each other and keep the particles separated
[11,12]. Besides, magnetic iron oxide particles should have surface
functional groups or ligands that have specific affinity to biomole-
cule to be separated efficiently [13]. Functional polymer coating
also renders the magnetic particles to have optimal surface bio-
compatible and upgraded properties desired. In addition, polymer
on the surface of the magnetic particles protects particles from sus-
ceptibility of leaching and oxidation, and improves particles colloi-
dal stability [14]. The encapsulation of pigments and other fillers
with polymers in emulsion is widely used in paint industries
[15]. In biomedical field, the role of polymers in the preparation
of magnetic polymer particles is generally to save the inorganic
part and to induce reactive chemical functions capable of interact-
ing with biological entities via chemical reaction or physical phe-
nomena [16].
Stimuli-responsive smart polymers that change their properties
with different environment such as pH, temperature, ionic

0021-9797/$ - see front matter Ó 2011 Elsevier Inc. All rights reserved.
doi:10.1016/j.jcis.2011.04.078
 M.M. Rahman et al. / Journal of Colloid and Interface Science 360 (2011) 556–564 557

strength, electrical field, magnetic field; have been used for the without purification. Triton X-405 was purchased from Sigma-
surface modification of magnetic particles to be tuned by external Aldrich. N-isopropylacrylamide (NIPAM) was obtained from Acros
properties. Pelton et al. [17] described the synthesis of temperature Organic and recrystallized from hexane and toluene. 2, 20 -azobis
sensitive poly (N-isopropylacrylamide) (PNIPAM) microgel and (2-methylpropionamidine) dihydrochloride (V-50) and AEMH
proved its volume phase transition at 32 °C. Other groups also were purchased from Wako and Acros Organics respectively. The
studied extensively on PNIPAM microgels preparation and charac- initiator V-50 was recrystallized from acetone. Magnetic emulsion
terization [18] that helped PNIPAM as a well-known temperature was purchased from AdemTech (Pessac, France). Deionised water
responsive polymer to make a coil-to-globule transition and was used in all experiments.
undergoes exquisite dehydration behavior at its lower critical solu-
tion temperature (LCST) [19,20]. As an emerging area, PNIPAM
2.2. Cross-linked magnetic polymer seed particle preparation
based magnetic polymer particles have been studied for different
applications such as catalysis [21,22], protein separation [23,24],
Core–shell structure and Janus like magnetic polymer particles
and bioprocess [25].
preparation from oil-in-water magnetic emulsion by the emulsion
Different ways have been reported for the preparation of PNI-
polymerization of Styrene (St) and cross-linker DVB were
PAM coated magnetic particles [26–29]. Shamim et al. [30] pre-
described in previous reports [31–33]. According to the above liter-
pared thermosensitive PNIPAM coated magnetic nanoparticles by
atures, cross-linked magnetic polymer particles were prepared
seed precipitation polymerization using thidiglycolic acid (TDGA)
using free radical emulsion polymerization of St/DVB (60/40%) or
and polymerizable 4-vinyl aniline coated iron oxide nanoparticles.
100% DVB in the presence of oil-in-water magnetic emulsion. The
They explained that the thiol group (–SH) and carboxylic group
polymerization was carried out according to a batch process in a
(–COOH) of TDGA can be attached with magnetic particles and
60 ml three necked double wall glass reactor equipped with a glass
amine group (–NH2) of vinyl aniline respectively. Pich et al. [29]
anchor type stirrer, a reflux condenser and a nitrogen inlet. The
prepared temperature responsive poly (N-vinylcaprolactam-co-
temperature was regulated at 70 °C by using a thermal bath.
acetoacetoxyethyl methacrylate) P(NVCL/AAEM) coated magnetic
50 ml of magnetic emulsion with solid content 4% (e.g. 2 g of dry
particles by the precipitation of iron oxide particles in the presence
form) was first stabilized with an aqueous solution of a non-ionic
of P(NVCL/AAEM). Magnetic particles were randomly scattered in
surfactant Triton-X 405 via serum replacement process performing
the polymer matrix prepared here that could be a shortcoming of
at least three separation/dispersion cycles. The Triton-X 405 solu-
this work.
tion was prepared at a concentration below the critical micelle
The core shell particle having both of the thermo-sensitivity and
concentration (CMC) 1 g L1. Magnetic emulsion was finally dis-
magnetic property could be of a great interest in separation bio-
persed in the Triton X-405 solution of the boiled and degassed
technology. Here, we have reported the preparation and character-
Milli-Q water, and introduced in the reactor and purged with nitro-
ization of thermosensitive core–shell magnetic P(NIPAM/MBA/
gen for 1 h under stirring (300 rpm) to remove oxygen. Then,
AEMH) composite particles starting from oil-in-water magnetic
720 ll of St with 480 ll DVB for MP-1, MP-2 and MP-3 particles
emulsion. TEM and light scattering results proved the morphology
or only 900 ll DVB for MP-4 particles was added in the reactor.
and volume phase transition behavior of the particles respectively.
The medium was kept under stirring at 300 rpm for 1 h at room
TGA result was found in a good agreement about magnetic content
temperature for the swelling of magnetic droplets by monomers.
in the hybrid composites prepared here. The novelty of this work is
Temperature was rapidly increased to 70 °C and 2 wt.% of KPS
to highlight a preparation method of perfect core–shell magnetic
(with respect to the weight of monomer and cross-linker)
polymer particles which have dual magnetic and highly tempera-
dissolved in 2 ml of boiled and degassed Milli-Q water was added
ture responsive properties. The core–shell magnetic particles can
in the monomer swollen magnetic emulsion at a temperature
be used as a solid support for easy magnetic separation of biomol-
70 °C. The polymerization was run at constant stirring (300 rpm)
ecules in biotechnology.
for 20 h.

2. Materials and methods 2.3. Synthesis of temperature responsive magnetic polymer particles

2.1. Materials DVB cross-linked magnetic polystyrene or only DVB cross-

Styrene (99%) and divinylbenzene (DVB) 80% were purchased
from Sigma–Aldrich and used after washing with 5% NaOH
solution. Potassium persulfate (KPS) and N,N0 -methylenebis
acrymamide (MBA) were obtained from Sigma–Aldrich and used
linked magnetic particles were washed three times with aqueous
solution of Triton X-405 (concentration below CMC) and poured
into the 60 ml reactor.100 ll of styrene for MP-1, MP-2 and
MP-3 or DVB for MP-4 was added to the seed magnetic particles
and stirred for 1 h at room temperature at 300 rpm for seed

Table 1
Polymerization recipe with different seed to monomers ratio for temperature sensitive magnetic polymer particles preparation; polymerization
temperature 70 °C, reaction volume 50 ml, stirring speed 200 rpm, time 20 h.

Sample Seed/monomer NIPAM (mol in MBAa (mol in AEMH (mol in V-50b (mol in Non-magnetic
(w/w) ratio feed)  101 feed)  103 feed)  103 feed)  103 polymerc (%)
MP-1 1.66 1.5 11.7 0.00 1.61 65
MP-2 2.50 0.97 7.78 2.97 1.16 51
MP-3 3.03 0.8 6.42 0.55 0.95 35
MP-4 3.53 1.7 7.78 4.8 1.27 18
a
MBA was taken 5 wt.% of NIPAM and AEMH.
b
Initiator was used 2 wt.% of the total monomer.
c
The amount of non-magnetic water soluble polymer was measured by gravimetrically and percentage is corresponding to total polymeri-
zation conversion that was found to be 100% in all runs after 20 h.
558 M.M. Rahman et al. / Journal of Colloid and Interface Science 360 (2011) 556–564

Scheme 1. Schematic representation of the temperature sensitive PNIPAM coated magnetic particles preparation starting from the oil-in-water magnetic emulsion.

particles swelling by monomer. A fraction of initiator solution 3. Results and discussion

(500 ll of V-50 dissolved in 2 ml) water was added first after rais-
ing temperature at 70 °C. After 1 h, the mixture of fixed amount
(detailed recipe was shown in Table 1) of NIPAM, AEMH, and
MBA dissolved in 8 ml water was added to the reaction system
and the remaining V-50 solution was added. The reaction was
carried out 20 h stirring at 200 rpm and the total volume was
kept at 50 ml. The total synthesis procedure of temperature
responsive PNIPAM coated core–shell magnetic particles is illus-
trated schematically in Scheme 1.
2.4. Characterization
Average hydrodynamic diameter (Dh) of the particles were mea-
sured using light scattering by a Zetasizer Malvern 3000HSA
(Malvern Instruments) at 90° angle. Very diluted sample measure-
ments were performed at ionic strength (103 M NaCl). The average
value of at least five measurements was taken at a given condition.
To measure the size at different temperature the particles were
equilibrated at a particular temperature for 6 min before measure-
ments. Zeta potential was measure by a Zetasizer Nano-ZS (Malvern
Instruments).
TEM images were obtained by a Phillips CM120 electron micro-
scope (CMEABG, University of Claude Bernard Lyon I). A drop of
well dispersed diluted sample was placed onto a copper grid
(200 mesh and covered with a formvar-carbon) and dried the sam-
ple at ambient at room temperature. Before taking the TEM image
the sample was diluted at least 10 times by water.
The chemical compositions of the dry particles and magnetic
iron oxide content were determined gravimetrically by using a
ZETZSCH thermogravimetry analyzer (TG209F1 Iris@ASC).
3.1. Synthesis of cross-linked magnetic seed polymer particles
TEM images presented in Fig. 1 clearly show different surface
morphology between Fig. 1A for oil-in-water magnetic emulsion
and, Fig. 1B and C for cross-linked magnetic polystyrene particles
and DVB cross-linked magnetic particles respectively. The results
observed here are compatible to the works done before wherein
core–shell [31,32] and anisotropic [33] magnetic polystyrene par-
ticles were prepared from magnetic emulsion. It is noticed that
the polymerization of St while DVB as cross-linker or only DVB
in the presence of magnetic ferrofluid droplets lead to make a
crosslink polymer shell on the surface of the magnetic seeds. The
use of 40 wt.% cross-linker DVB during Styrene polymerization or
100% DVB enhances to form a homogeneous polymeric shell onto
the magnetic core. The increase of hydrodynamic diameter mea-
sured by dynamic light scattering from magnetic emulsion drop-
lets (300 ± 10 nm) to cross-linked magnetic polymer (350 ± 10) is
around 50 nm.
TEM images also reveal that both of the monomer composition
St/DVB (60/40 wt.%) and 100% DVB were consistent to prepare
magnetic core and cross-linked polymer shell particles that can
be used as seed for further functionalization.
3.2. Synthesis of temperature sensitive polymer coated magnetic
particles
A series of polymerization were carried out according to the
recipe mentioned in Table 1. As a template, seed cross-linked
magnetic polystyrene or only DVB cross-linked particles were

Fig. 1. TEM images of (A) oil-in-water magnetic emulsion (B) DVB cross-linked magnetic polystyrene particles and (C) DVB cross-linked magnetic particles.
 M.M. Rahman et al. / Journal of Colloid and Interface Science 360 (2011) 556–564 559

Fig. 2. TEM image of (A) temperature responsive magnetic particles MP-1for which seed/ monomer (w/w) ratio was 1.66 during polymerization, (B) sample particles MP-2
seed/monomer (w/w) ratio 2.5 and (C) MP-3 seed/ monomer (w/w) ratio 3.03.

Fig. 3. TEM images (A and B) of temperature sensitive poly(NIPAM) coated magnetic particles MP-4 for which only DVB cross-linked seed/monomer (w/w) ratio was 3.53
during polymerization.

swelled with small amount of styrene or DVB to increase the
efficiency of the incorporation of temperature sensitive P(NIPAM/
MBA/AEMH) shell onto the seed particles.
The amount of water soluble non-magnetic polymer were
determined by gravimetrically and calculated in percentage from
the polymerization conversion. It is observed that the use of higher
quantity of water soluble monomers NIPAM, MBA and AEMH with
respect to seed increases aqueous phase non-magnetic polymer
formation rather than to be incorporated on the surface of the seed.
It shows that the seed/monomers ratio 1.66 gives more than 60%
non-magnetic water soluble polymer P(NIPAM/MBA/AEMH) in
the aqueous medium while the ratio 3.53 reduces it at below
20%. Morever, in the later case there might have cross-linker effect
because of seed swelling with DVB. It is worth mentioning that our
aim was to prepare perfect core–shell morphology of the final
magnetic particles having cationic polymeric shell onto a nega-
tively charged seed magnetic polymer particles. TEM images of
Figs. 2 and 3 represent that the seed/monomers (NIPAM, MBA
and AEMH) ratio in the second stage polymerization could play
an important role on the morphology of the particle and in partic-
ularly on the formation of the non-magnetic polymers. Hydrody-
namic diameter of magnetic particles increases being more
encapsulated with PNIPAM with decreasing the amount of non-
magnetic polymers except the particle MP-1. It is also apparent
in Fig. 2 that magnetic PNIPAM particles MP-1 and MP-2 are not
well dispersed as found in the TEM of MP-3 and especially in
MP-4 (Fig. 3); however, all particles are core–shell type morphol-
ogy. This can be attributed to the water soluble PNIPAM hydrogel
560 M.M. Rahman et al. / Journal of Colloid and Interface Science 360 (2011) 556–564
Fig. 4. Thermogravimetric results of magnetic emulsion, cross-linked magnetic seed polymer and PNIPAM coated magnetic particles (MP-2, MP-3 and MP-4).
that might be involved in some extent bridging formation between
the hydrophilic shells present on the surface of the magnetic seed
polymers. The bridging formation may be the cause of larger parti-
cles size of MP-1 rather than MP-2, MP-3 and MP-4.
3.2.1. Core–shell morphology
The morphology of composite polymer particles prepared by
second stage seed emulsion polymerization depends on the differ-
ent factors such as initiator [34] surface tension, molecular weight
of second stage polymers, degree of monomer swelling, chain
transfer agent, monomer addition [35–37]. In addition to this,
the amount of monomer with respect to seed polymer also has a
great impact onto the morphology of the final particles [34].
According to Stutman et al. [38] who studied Poly (butylacry-
late)/PS, the phase separation in the monomer rich surface layer
of the seed particles and the capture of a secondary crop of polysty-
rene particles from the aqueous phase by the seed latex particles
controls the morphology of composite latex particles. Cho and
Lee demonstrated [39] that (PMMA/PS) the ionic end groups’ that
are present on the polymers interaction for anchoring and the vis-
cosity within the particles are vital factors to control morphology.
The seed latex particles swollen to some level with monomer and
polymerization within the particles, in this case, phase separation
will occur through nucleation and growth process. In our system
cross-linked magnetic polymer seed particles were first swelled
with St or DVB. After swelling St and DVB diffused in the
cross-linked polystyrene (for MP-1, MP-2 and MP-3) and DVB
cross-linked (for MP-4) shell of the magnetic seed polymer and
polymerization initiated on the surface of the seed particles those
could be the nucleation center for particles growth for water solu-
ble P(NIPAM/MBA/AEMH) and initiator V-50. Considering the seed
particle as particle nucleation and growth center, lower seed to
monomer ratio led to form water soluble non-magnetic polymers,
eventually being precipitated at reaction temperature 70 °C (above
LCST), due to the lack of seed particles surface. That is why, seed
magnetic particles are not homogenously encapsulated by PNIPAM
in MP-1, MP-2 while MP-3 is better encapsulated. However, same
seed particles were used for functionalization. On the other hand,
magnetic seed particles are perfectly coated by PNIPAM in the
MP-4 particles. The number of seed particles (in other way amount
of seed particle) should be considered for core–shell structure of
particles. Those are the responsible to capture the PNIPAM chains
from the reaction medium.
Moreover, any colloidal system prefers to organize itself to
reach the most thermodynamically favoured state corresponding
to the minimum free energy (DG) and lowest interfacial free en-
ergy of system. It can be assumed that driving force for the core–
shell morphology of these magnetic PNIPAM particles is originated
from a thermodynamic equilibrium between the second polymer
phase and the cross-linked magnetic polymer seed particles. The
lowest final Gibbs free energy leads the particles’ favoured mor-
phology. The Gibbs free energy is related to enthalpy and entropy.
If the enthalpy changed is the same regardless of morphology and
considering that the particles are macroscopic comparison to small
molecules to neglect entropy. Then, surface energy is the key
parameters to determine the particles morphology. The polymer
polarity, surfactant type and concentration, initiator end groups,
can influence the interfacial energy and finally on the particles
morphology [40]. From the above discussion and TEM observation
in Fig. 3, we may conclude that the parameters used in the reaction
system for MP-4 are enabled to push the system at thermodynam-
ically favoured and low free energy state. As a consequence, parti-
cles tend to be in core–shell structure. Thus, it can be a method of
choice for the preparation of perfect core–shell magnetic polymer
particles having temperature responsiveness. Fig. 3 also shows the
well dispersion without any aggregation of MP-4 particles.
3.3. Thermogravimetry analysis
The amount of P(NIPAM/MBA/AEMH) bound on the surface of
the prepared particles is estimated from the percentage of weight
loss from the TGA curve represented in Fig. 4. The major weight
loss occurs in the range of 350–450 °C and tends a plateau after
500 °C. As expected, the amount of residual mass (mostly magnetic
content and little amount ash) about 60% in cross-linked magnetic
seed particles is lower than that 82% of magnetic emulsion. These
values are also obviously higher than those of P(NIPAM/MBA/
AEMH) coated magnetic particles. Every step of polymerization
incorporates polymers onto seed particles and lowers magnetic
content in order to increasing organic fraction in the magnetic par-
ticles. After degradation of organic part, residual magnetic mass of
the particles MP-2 and MP-3 are almost equal around 55%. Since
this value is not far from the value of seed magnetic polymer
(60%). Therefore, it indicates the lower encapsulation efficiency
of seed by P(NIPAM/MBA/AEMH). On the other hand, residual mass
value (44%) of MP-4 particles supports higher encapsulation
 M.M. Rahman et al. / Journal of Colloid and Interface Science 360 (2011) 556–564 561

Fig. 5. (A) hydrodynamic diameter and (B) swelling ratio Sw = V/Vc of temperature responsive magnetic particles as a function of temperature. Measurements were performed
at 1 mM NaCl solution.

efficiency by thermosensitive polymer. All of the TGA results are to the high content water soluble non-magnetic polymer formation
compatible to results found in non-magnetic polymer determina- that leads low incorporation of PNIPAM on the surface of the seed
tion and TEM images of corresponding particles prepared. The magnetic particles. As it has been found before in the TEM images,
magnetic content in the final particles is sufficient to be a very the thickness of thermosensitive polymer shell in MP-4 is more
good magnetic sensitive in an external magnetic field and can be dense and homogeneous rather than MP-3. But the volume phase
separated quickly from the separation medium without difficulties. change i.e. swelling ratio is high for MP-3. This can be explained
This is because the produced magnetic polymer particles can be in such way that a fraction of DVB that was used for seed magnetic
used for biomolecules such as nucleic acids separation technology. particles swelling could be remain in the aqueous phase and con-
tribute to MBA to increase the cross-linker concentration. More-
3.4. Colloidal and temperature responsive properties over, there might have an effect of the amount of functional
monomer AEMH what was 4.8103 mole in MP-4 polymerization
3.4.1. Temperature effect on hydrodynamic diameter recipe, while it was 0.55103 mole for MP-3.
The hydrodynamic diameter as well as swelling ratio of P(NI- Fig. 6 shows the normalized hydrodynamic diameter of parti-
PAM/MBA/AEMH) coated cross-linked magnetic particles as a func- cles and indicates that all particles have phase transition of size
tion of temperature are given in Fig. 5A and B respectively. The with increasing temperature. Major transition is observed in the
swelling ratio (Sw) was calculated by the following equation [41]. range 25–40 °C depending on the particles due to the collapse of
the PNIPAM shell at higher temperature. It should be mention that
Sw ¼ V=V c
there is no marked difference in the normalized phase transition
where V and Vc is the volume of particle at a given temperature and among different particles MP-1, MP-2, MP-3 and MP-4. However,
collapsed state (in this case volume at 50 °C) respectively. The par-
ticles MP-1, MP-2, MP-3 and MP-4 are found to show decrease in
hydrodynamic diameter upon heating with variation of tempera-
ture at 1 mM NaCl solution.
As expected, this behavior comes from the shrinkage of the
P(NIPAM/MBA/AEMH) shell of the magnetic particles. It is also
mentionable that PNIPAM hydrogel chains collapse above it lower
critical solution temperature (LCST), eventually being dehydrated
by expelling water molecule outside of the hydrogel shell, which
result in shrinkage and increases surface charge density [42]. At
temperature below LCST, hydrogen bonds between hydrophilic
groups of polymer chains are dominant, i.e. increase of water
absorption. These bonds become weak and hydrophobic interac-
tions between polymer chains become stronger when temperature
is elevated over the LCST. This phenomenon is reversible and
hydrogel return in his swollen state when temperature is lowered
below the LCST. The use of functional monomer AEMH during
polymerization with NPAM renders amino (–NH2) functional
groups in the chains of the cross-linked PNIPAM shell. These amino
groups can play an important role to differ volume phase transition
temperature (VPTT) and size of the particles at different pH. It is
observed that the significant phase transition of all particles pre-
pared here except the particles MP-2 which show the least volume Fig. 6. Normalized hydrodynamic diameter of thermo-sensitive magnetic particles
change upon heating from 20 °C to 50 °C. This result might be due as a function of temperature.
562 M.M. Rahman et al. / Journal of Colloid and Interface Science 360 (2011) 556–564

Fig. 7. The hydrodynamic diameter of poly (NIPAM/MBA/AEMH) coated magnetic particles as a function of temperature at different pH (ionic strength 1 mM NaCl); (A) MP-2
(seed: monomer (w/w) ratio 2.5), (B) MP-3 (seed: monomer (w/w) ratio 3.03) and (C) MP-4 (seed: monomer ratio was 3.53).

the difference has been seen before in Fig. 5A and B in terms of
hydrodynamic diameter and swelling ratio.
It is better to mention that the small change of hydrodynamic
diameter (Dh) of the particles MP-2 is might be due to the use of
higher amount hydrophilic monomer aminoethylmethacrylate
hydrochloride (AEMH) mention in Table 1 in the polymerization
recipe enhances water soluble polymers formation and conse-
quently low shell thickness (i.e. low Dh of the magnetic particles).
As a result small change of the Dh with increasing temperature is
observed in Fig. 5A and B and Fig. 6 comparing to other particles
prepared here. Moreover, the presence of hydrophilic AEMH in
poly(NIPAM) backbone, reduces temperature responsiveness of
the shell that might be another cause of little Dh change of MP-2
particles versus temperature.
3.4.2. Effect of pH on hydrodynamic diameter at different temperature
The shrinkage behavior of thermo-sensitive magnetic polymer
particles except the particle MP-1 was studied at different pH.
Fig. 7 shows that there is massive effect of particles’ dispersion
medium pH on the hydrodynamic diameter change as function of
temperature. The reason behind the low difference in size at differ-
ent pH can be incurred to cross-linker which can arrest the mobil-
ity of hydrophilic shell and low thickness of hydrogel shell.
However, commonly, for all particles, size is found to be increased
in a little extent (10–50 nm) at acidic pH than that of at basic pH. It
is also noticed that this type of size change trend is almost same
regardless of pH. Acidic pH value of the dispersion medium leads
to have more protonation of amino (–NHþ 3 ) groups derived from
AEMH and amidine groups coming from initiator V-50 present in
the particles surface [43]. These protonated groups enhance the
charge density of particle’s surface and, thus PNIPAM chains tend
to be far away from each other, eventually, undergoing more elec-
trostatic repulsion. On the other way, higher electrostatic repulsion
between PNIPAM hydrogel chains increases osmotic pressure of
the polymer shells. This osmotic pressure is balanced by PNIPAM
shell swelling [44,45]. It is not apparent markedly above 40 °C tem-
perature as consequence of hydrophobic nature of PNIPAM shell of
the magnetic polymer particles.
3.4.3. Temperature effect on zeta potential
Fig. 8 shows the dependence of electrophoretic mobility con-
verted to zeta potential on temperature. Zeta potential was mea-
sured at 1 mM NaCl solution as a function of temperature. The
zeta potential of all particles (MP-1, MP-2, MP-3 and MP-4) was
found to be increased with increasing temperature. By the combi-
nation of Helmoltz’s and Smoluchowski’s formula [46] regarding
 M.M. Rahman et al. / Journal of Colloid and Interface Science 360 (2011) 556–564
Fig. 8. Zeta potential as a function of temperature (measurements were carried out
at 1 mM NaCl solution and different temperature 20–50 °C).
charge distribution model, it can be possible to make a relation be-
tween electrophoretic mobility (le) surface charge density (r) and
hydrodynamic diameter (2rh) as follows:
le  Ne=4pgjr2h
where N is the charged groups’ number, e charge of an electron, g is
the viscosity of the medium, j is the reciprocal Debye length.
According to the above equation, the electrophoretic mobility of
particles increases with decreasing hydrodynamic diameter (2rh).
With elevating temperature, the thermosensitive PNIPAM shell of
magnetic particle undergoes collapse state which results to uphold
surface charge density of particles. As a result, zeta potential of par-
ticles becomes higher at higher temperature that also proves the
temperature responsiveness of the prepared particles.
The zeta potential changing trend of MP-1 is sharp than that of
other particles. This can be attributed to the absence of AEMH in
the polymerization with NIPAM. Because, the presence of AEMH
which renders hydrophilic amino functional groups in the PNIPAM
chains and as a result, PNIPAM shell becomes less thermally sensi-
tive. For the remaining particles those contain AEMH, phase transi-
tion in terms of zeta potential is noticed as wider than that of the
first one. In addition, the absolute values of zeta potential and VPTT
for each particle are not fully argued in relation to the composition
of the PNIPAM-based shell layer. Actually, to find out exactly same
phase transition in terms of Dh of PNIPAM shell and zeta potential
is somewhat difficult when highly rigid magnetic core is present in
the particles. The magnetic content makes the particles phase tran-
sition behavior different from classical microgel.
4. Conclusion
Temperature responsive PNIPAM shell containing magnetic par-
ticles have successfully been prepared by seed emulsion polymeri-
zation. DVB cross-linked magnetic polystyrene or DVB cross-linked
magnetic particles were obtained by the emulsion polymerization
of DVB and St in the presence of oil-in-water magnetic emulsion
as seed. Cross-linked magnetic particles were further subjected to
functionalize with NIPAM, AEMH as principal and functional mono-
mer respectively while MBA and V-50 were used as cross-linker and
initiator accordingly. Results indicate that seed to monomer ratio
more than three can be a better choice for inducing temperature
563
responsive PNIPAM on the seed particles by precipitation polymer-
ization and to have very good magnetic core polymer shell of the
composite particle. TEM micrographs have displayed the morphol-
ogy of the magnetic particles. For all latexes, hydrodynamic
diameter decreasing and zeta potential increasing with increasing
temperature confirm the temperature responsive and surface
charge properties of the magnetic particles prepared here. It is also
found that the volume phase transition temperature can be affected
by the pH of the particles dispersion medium. The use of functional
monomer AEMH and initiator V-50 provide cationic surface prop-
erty that was obtained from zeta potential measurements of parti-
cles. Last but not least, we hope that perfect core–shell composite
particles which contain both magnetic and temperature responsive
properties have been successfully prepared. These cationic hairy
shell containing magnetic particles can serve for nucleic acid sepa-
ration and purification in biotechnology.
Acknowledgments
This work has been achieved in the frame of Advanced Diagnos-
tics for New Therapeutic Approaches (ADNA), a program dedicated
to personalized medicine, coordinated by Institut Mérieux and
supported by research and innovation aid from the French public
agency, OSEO.
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