a n a l y t i c a c h i m i c a a c t a 6 0 8 ( 2 0 0 8 ) 197–203

available at www.sciencedirect.com

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

Analytical followup of the gamma initiated synthesis

of a molecularly imprinted polymer

Zoltán Zsebi a , Viola Horváth b , Ágnes Sáfrány c , George Horvai a,∗

a Budapest University of Technology and Economics, Department of Inorganic and Analytical Chemistry, Szt. Gellért tér 4,
H1111 Budapest, Hungary
b Hungarian Academy of Sciences, Research Group of Technical Analytical Chemistry, Szt. Gellért tér 4, H1111 Budapest, Hungary
c Institute of Isotopes, Hungarian Academy of Sciences, P.O. Box 77, H1525 Budapest, Hungary

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

Article history: A molecularly imprinted polymer (MIP) for the template phenytoin has been prepared by
Received 25 September 2007 gamma initiated copolymerization of methacrylamide and ethylene glycol dimethacrylate.
Received in revised form The progress of polymerization was studied by measuring the monomer conversions and
10 December 2007 the template binding properties of the resulting polymers, respectively. The consumption
Accepted 12 December 2007 rate of the two monomers showed different course. There was no difference observed in
Published on line 23 December 2007 the polymerization rates of the MIP and the control polymer (NIP). The template binding
properties of the MIP and the NIP changed considerably with the progress of the polymer-
Keywords: ization process and became similar to those of the thermally initiated polymers after full
Molecular imprinting conversion.
Gamma polymerization © 2007 Elsevier B.V. All rights reserved.
Radiation polymerization
Conversion rates

1. Introduction monomers around the target (template) molecules with the

In the last two decades the research of molecularly imprinted
polymers (MIP), a new class of selective sorbents, has become a
popular area in materials science. Many applications of these
polymers have been reported due to their special molecular
recognition and binding properties. MIPs are suitable sorbents
for solid phase extraction (SPE) [1–3] and liquid chromatogra-
phy, and as artificial antibodies in binding assays [4]. They have
been also successfully used for chiral separation [5], mem-
brane separation [6] and as chemical sensors [7].
MIPs are highly crosslinked, porous, synthetic polymers
with large surface area and suitably positioned and oriented
functional groups responsible for the recognition of target
molecules. The chemical and steric selectivity of MIPs is
developed in the polymerization process by fixing functional
help of a large excess of a crosslinking monomer.
The most widely used method to create MIPs is non-
covalent imprinting [1,8]. In this process, the functional
monomers and the templates form complexes at first. This
is followed by the polymerization step. Finally the resulting
polymer is ground (if necessary) and washed to remove the
template molecules.
Imprinted polymers are usually synthesized by free radical
polymerization using either thermal or UV initiation. Thermal
polymerization is a very simple, low cost method to prepare
polymers. No special equipment is needed, but the elevated
temperature (typically about 60 ◦ C), which is an essential con-
dition of the process, does usually not favor the formation of
monomer–template complexes. On the other hand the tem-
perature of the UV polymerization can be set freely, but the

∗
Corresponding author. Tel.: +36 1 463 1480; fax: +36 1 463 3408.
E-mail address: george.horvai@mail.bme.hu (G. Horvai).
0003-2670/$ – see front matter © 2007 Elsevier B.V. All rights reserved.
doi:10.1016/j.aca.2007.12.014
198 a n a l y t i c a c h i m i c a a c t a 6 0 8 ( 2 0 0 8 ) 197–203

homogeneous illumination of the polymerization mixture is

problematic. This fact and the problems with the dissipation
of reaction heat may cause difficulties in scaling-up. In both
the thermal and the UV methods the polymerization is started
by reactive radicals, generated by thermal or UV homolysis of
an initiator.
Irradiation of the mixture of monomers with ionizing radi-
ation, such as gamma- or electron beam, is a way to initiate
free radical polymerization without any added initiator. In this
case the active species is formed by the interaction between a
compound in the polymerization mixture (monomer, solvent,
etc.) and the electromagnetic or electron beam. The advan-
tages of high-energy irradiation induced polymerization are
the wide range of applications, ease of operations at room
temperature, the easily controllable rate of polymerization,
and the ability to irradiate large samples homogenously. In
polymer chemistry the most important application areas of
gamma initiation are the polymer modifications by grafting,
and the curing and crosslinking of polymers [9]. The gamma
irradiation polymerization is also an applicable technique for Fig. 1 – Chemical structures of the co-monomers: ethylene
creating polymeric materials with rigid structure and high glycol dimethacrylate (a), methacrylamide (b) and the
surface area, and for controlling the shape, size and porosity template phenytoin (c).
of the products. These optimized products then can be used
successfully as stationary phase for chromatography [10,11].
Although gamma irradiation is not a common technique in dose was determined by using ethanol chlorobenzene dosime-
research labs, its advantages have lead to its wide use in prac- ter [18].
tical applications. HPLC measurements were made on a PerkinElmer (PE)
There have been only a few attempts to use gamma radi- Series 200 HPLC system equipped with a pump, programmable
ation in molecular imprinting. Uezu et al. [12] and Biju et autosampler and a variable wavelength UV/vis detector,
al. [13] have studied the effect of gamma post-irradiation PerkinElmer Series 600 interface and PerkinElmer Turbochrom
on the selectivity of ion imprinted polymers (IIP). Kala et Navigator 6.1.2.0.1:D19 software. All HPLC measurements
al. [14] synthesized erbium(III) IIP from a whole series of were made in duplicate and the average of the two values was
monomer–crosslinker pairs with gamma irradiation. Some used in the calculations.
studies of preparing imprinted polymers with organic tem-
plate molecules – thus (±)-menthol [15], testosterone [16] and 2.2. Materials
cholesterol [17] – have been also reported. None of these
authors has studied the progress of the polymerization pro- Ethylene glycol dimethacrylate (EDMA, Fig. 1a, Fluka, purum,
cess. ≥97%) was distilled from the inhibitor under reduced pressure.
In our work phenytoin imprinted poly(methacrylamide-co- Methacrylamide (MAAM, Fig. 1b, 98%) and phenytoin (Fig. 1c,
ethyleneglycol-dimethacrylate) polymers have been prepared 5,5-diphenylhydantoin, 99%) were purchased from Aldrich
using gamma irradiation. The progress of polymerization and were used without any further purification. Tetrahydrofu-
and of the creation of binding sites has been studied by ran (UV-IR-HPLC grade) and acetonitrile (HPLC-gradient grade)
making and investigating different batches of the polymers were purchased from Panreac Chı́mica S.A. Both solvents were
with different doses of irradiation. In contrast to UV and dried on 3 Å Molecular Sieve Deperox/Dehydrate (Fluka).
thermal initiation the effect of gamma irradiation can be
expected to be homogeneous in the whole sample volume. 2.3. Synthesis of polymers
The effects of the applied irradiation dose have been followed
by measuring the conversion rates of each monomer and by For polymer synthesis a process developed by Lanza et al. [19]
measuring the phenytoin rebinding properties of the result- and called miniMIP technique was used. This method is appli-
ing polymers. The polymers made by gamma polymerization cable for screening properties of a large number of samples
have also been compared with those obtained by thermal with many repeated experiments.
polymerization. Two mother solutions were prepared: one with template
(for the MIP) and one without template (for the control
non-imprinted polymer, the NIP). The MIP mother solution
2. Experimental contained 133 mg (0.53 mmol) phenytoin template, 181 mg
(2.12 mmol) methacrylamide (MAAM) functional monomer
2.1. Instrumentation and 2.11 g (10.6 mmol) ethylene glycol dimethacrylate (EDMA)
crosslinking agent dissolved in a volumetric flask of 5 mL
The irradiation was done on the panoramic 60 Co gamma volume by a solvent mixture of tetrahydrofuran and acetoni-
source (type SLL-01) of the Institute of Isotopes Co. Ltd. The trile in 3:7 mass ratio. The molar ratio between the template,
 a n a l y t i c a c h i m i c a a c t a 6 0 8 ( 2 0 0 8 ) 197–203 199

the functional monomer and the crosslinker was 1:4:20. The

mother solution of non-imprinted control polymer (NIP) was
3. Results and discussion
prepared in the same way but without template. The mother
solutions were purged with argon for 15 min in order to Phenytoin imprinted and non-imprinted polymers were pre-
eliminate the oxygen, which can effect the polymerization pared by gamma radiation initiated polymerization. The
reaction. Then 120 ␮L aliquots of the mother solutions were polymers irradiated with low doses of gamma ray (less than
pipetted into 1.5 mL glass vials, which had been purged with 10 kGy) were transparent and gel-like. These polymers lost
argon previously. After the distribution of solutions the vials more than half of their volume during the drying process
were purged with argon again, and finally were sealed with (according to visual observation), and swelled again after
PTFE/silicon rubber septa. adding the acetonitrile solvent. The degree of the collapse
The vials were subjected to gamma irradiation from a 60 Co and the transparency of the polymers decreased with increas-
source at a dose rate of 12 kGy h−1 . The total absorbed dose of ing dose of irradiation. The polymers prepared with 10 kGy or
gamma radiation was controlled by varying the time of the higher dose were hard, white blocks without visible swelling.
irradiation. The applied reaction times were 20, 30, 40, 50,
75 and 100 min, respectively, thus the absorbed total doses 3.1. Co-monomer conversion studies
were 4.0, 6.0, 8.0, 10, 15 and 20 kGy, respectively. These calcu-
lated dose values were in all cases checked by chlorobenzene After the polymerization, the non-reacted monomers were
dosimeter attached to the sample. dissolved in a volume of acetonitrile and the concentration
Every polymerization experiment was made with five par- of monomers was measured by HPLC. The conversion of
allels, i.e., in five vials, each with the same composition and monomers was defined as (1 − M)/M0 , where M and M0 are the
the same irradiation dose. All reported data are averages of starting- and the measured unreacted amounts of the studied
the five parallel results with the standard deviation shown in monomers, respectively. Fig. 2 shows the calculated methacry-
the figures by bars. lamide and ethylene glycol dimethacrylate conversion values
against irradiation time (or dose of gamma ray). The corre-
2.4. Measuring the monomer conversion sponding data of the MIPs and NIPs are shifted along the
time-axis with +1 and −1 min (0.2 kGy), respectively, in order
After the respective irradiation (polymerization) times 1 mL of of easier visibility.
acetonitrile was added to each vial. After an overnight period There is no significant difference between the monomer
the amount of dissolved unreacted monomers was measured conversions observed with the MIP and NIP polymers, respec-
by HPLC. The percentage of the monomer conversion was cal- tively (Fig. 2, full vs. empty symbols). This means that the
culated from the ratio of the amount of consumed monomer presence of the template phenytoin has no influence on the
and the starting amount of the corresponding monomer. rate of the polymerization.
On the other hand, the polymerization rate of the two co-
2.5. Rebinding measurement monomers is not equal. The full conversion of EDMA occurs
much faster than MAAM conversion. The EDMA conversion
The polymers need to be carefully cleaned before rebinding versus time is almost linear in the first 50 min (10 kGy) of poly-
experiments. To eliminate even traces of the template and the
unreacted monomers from the polymers, these were washed
with a 10:90 (v/v) mixture of acetic acid and methanol until
no traces of phenytoin were detected. The purity of washing
solvents was checked with HPLC/UV. The purified polymers
were dried in vacuum at room temperature.
The template rebinding properties of the prepared MIPs
and NIPs were determined by the batch equilibration method.
The mass of each cleaned and dried polymer sample was mea-
sured and a 1 mM solution of phenytoin in acetonitrile was
added to each sample. The ratio of the volume of solution
to the mass of the polymer was always 6 ␮L mg−1 . Then the
samples were placed into a thermostat of 25 ◦ C for 24 h. The
equilibrium concentration of the template in the supernatant
was measured by HPLC. The specific adsorption (q, mmol kg−1 )
on MIPs and NIPs was calculated as the ratio of the adsorbed
molar amount of template to the mass of the polymer. The
distribution ratio (D, dm3 kg−1 )) of the template was calcu-
lated as the ratio of the specific adsorption and the equilibrium
solution concentration of the template, respectively. Imprint-
ing factor (IF) was used for the comparison of the adsorption
properties of imprinted and non-imprinted polymers. IF was Fig. 2 – Conversion of MAAM and EDMA vs. time and dose
calculated as the ratio of the respective template distribution of irradiation. For better visibility the points have been
ratios on the MIP and the NIP. shifted by 1 min to the left (NIP) or to the right (MIP).
200 a n a l y t i c a c h i m i c a a c t a 6 0 8 ( 2 0 0 8 ) 197–203

(dose) may cause the differences in the template rebinding

properties of polymers shown in the next section.

3.2. Rebinding studies

The template rebinding properties of imprinted and non-

imprinted polymers were characterized by the phenytoin
distribution ratio (D, dm3 kg−1 ) between the solid polymer and
the solution. D was defined by the equation:

(c0 − ceq )V
D= ,
ceq m

where c0 is the original (1 mM) and ceq is the equilibrium con-

Fig. 3 – Molar fraction of MAAM in polymers (as calculated
from the monomer consumptions) vs. time and dose of
irradiation.
centration of phenytoin in mmol dm−3 , V is the volume of
added solution in cm3 , and m is the mass of the polymer in
g. The distribution ratios for polymers made with the differ-
ent doses of gamma rays are shown in Fig. 4a. The first, and
the most important conclusion seen from the diagram is, that
the distribution ratio of the template is significantly higher
on imprinted polymers than on non-imprinted polymers. In
the first period of polymerization (up to 6–8 kGy dose) this
is very interesting, because the rigid structure of the func-
tional groups probably could not be formed yet. It can be
seen also, that phenytoin binding increases with the increase

merization, up to 80–90% conversion of this monomer. Then

the conversion of EDMA reaches the 100% level before 75 min
(15 kGy dose of gamma ray). In the case of MAAM, there are
three periods in the conversion: a fast, a slow and a subsequent
fast period. The first fast period is finished prior to the gelation
point, which is before the first measured point at 20 min (4 kGy
dose). Then there is a slow period followed by a second period
of faster conversion. In this last period the rate of MAAM con-
version comes close to the rate of EDMA conversion. This fast
period lasts until the complete conversion of methacrylamide
(about 75 min, 15 kGy dose).
Because of the different rates of the polymerization
of monomers, the composition of polymers changes with
time during the polymerization. Fig. 3 shows the calculated
molar fraction of methacrylamide in the imprinted and non-
imprinted polymers versus the polymerization time and the
dose. It should be noted that the polymer compositions were
calculated from the consumption of the individual monomers
from the polymerization solution. This calculation assumes
that all the disappeared monomers were built into the poly-
mer comprising the solid MIP or NIP. It is possible, however,
that some amount of shorter oligomeric chains is also formed
and remains in solution but goes undetected by our HPLC
method. This possibility could be excluded for all but the
first two gamma doses (4 and 6 kGy, or 20 and 30 min, respec-
tively) because the mass of polymer obtained was always close
(within 8%, despite difficulties in its measurement) to the
value calculated from the monomer consumption (data not
shown here).
As shown in Fig. 3, the molar ratio of MAAM in the polymers
remains about 0.11 up to the 10 kGy dose and then increases
to the final value of 0.17 (which agrees with the composition
of the original polymerization mixture). The composition of Fig. 4 – Distribution ratio (a) and imprinting factor (b) of
the polymers changes in the same way both in MIPs and in phenytoin on NIPs and MIPs at different doses of
NIPs. These changes in the polymer composition with time irradiation.
 a n a l y t i c a c h i m i c a a c t a 6 0 8 ( 2 0 0 8 ) 197–203
Fig. 5 – Distribution ratio and imprinting factor of
phenytoin on thermally and gamma polymerized NIPs and
MIPs, respectively, after full conversion of the monomers.
of the dose of irradiation up to 15 kGy, and then it becomes
constant.
For the description of the effect of the template on
the rebinding properties of polymers, the imprinting factor
(IF = DMIP /DNIP ) was calculated. The imprinting factors of poly-
mers prepared with different doses of irradiation, are shown
in Fig. 4b. The IFs also increase with the irradiation dose up to
15 kGy.
The rebinding properties were also compared between the
polymers prepared by gamma initiation at maximum dose and
by thermally initiated polymerization. The thermally polymer-
ized MIP and NIP were prepared previously at 60 ◦ C with added
initiator as described elsewhere [19]. The distribution ratios of
phenytoin and the imprinting factors, measured on the poly-
mers prepared with the two different methods are shown in
Fig. 5. It is seen, that the imprinting factor on ‘gamma MIP’ is
higher than on ‘thermal MIP’, but this difference is not signifi-
cant because of the large standard deviation of data measured
on ‘thermal MIP’. On the other hand there is significant dif-
ference between the distribution ratios measured on the two
kinds of polymers. This means that the gamma polymerized
MIP can adsorb almost two times more phenytoin under these
conditions than the MIP prepared by thermal polymerization.
3.3. Gamma initiation as an alternative for making
and investigating MIPs
The details of the thermal or UV polymerization processes
leading to MIPs have not yet been well clarified. The exact
topology and chemical constitution of the selective binding
sites of MIPs is little known, particularly for non-covalent
MIPs. These are good reasons to investigate alternative means
of polymerization and see if the resulting MIPs are different
from the thermal or UV polymerized MIPs. Gamma irradia-
tion has been frequently used to make polymers, including
porous polymeric adsorbents similar to the control poly-
mers of MIPs, i.e. NIPs. Gamma polymerization has certain
advantages: there is no need for initiator, initiation occurs
201
homogeneously even in a thick block of sample and no ele-
vated temperatures are needed as in thermal initiation. This
means that the rate of initiation in gamma polymerization
can be made more uniform in space and time than with the
alternative methods. This may be advantageous for the MIP
properties and it is certainly an advantage for kinetic studies
like in this paper. Potential disadvantages of gamma irradia-
tion include partial decomposition of the template and of the
polymer itself, due to radiolysis.
Earlier work using gamma irradiation for making MIPs is
scarce and not sufficiently detailed. Most of these studies
deal with special applications: preparation of “ion imprinted
polymers” or post-irradiation of MIPs made by other means.
Sreenivasan and Milojkovic describe non-covalent MIPs pre-
pared by gamma irradiation. The polymers of Sreenivasan
appear to be quite good ones. He also compared the gamma
polymerized polymer’s binding properties with those of the
UV and thermally polymerized MIPs for the same template.
Interestingly, there was very little difference between the UV
and gamma polymerized MIP, while both were better than the
thermal MIP. The MIP of Milojkovic et al. was only marginally
different from the control polymer (NIP).
None of the aforementioned studies discussed the role of
gamma irradiation dose or time. We have found in the present
work, that the dose variable has profound influence on the
physical and template binding properties. MIPs and NIPs pro-
duced with less than 10 kGy were swellable in acetonitrile.
The extent of swellability decreased with increasing doses
and no swelling was observed visually above 10 kGy. Template
binding increased with the dose up to a certain limit. The
imprinting factor was also increasing with the irradiation dose
(Fig. 4b). Irradiation beyond 15 kGy appears not to improve
the analytical properties but may gradually degrade the poly-
mer. Therefore an optimum dose of irradiation is indicated. It
remains unclear if the other authors using gamma initiation
employed the optimal dose for their respective polymers.
3.4. The relationship between the polymerization
process and the properties of MIPs
Our study of the dose effects revealed also an interesting dis-
parity in the rate of conversion of the two monomers MAAM
and EDMA (Fig. 2). At the beginning of the polymerization (i.e.,
at low irradiation dose or short irradiation time) the conver-
sion rates of the two monomers are similar. Later, however, the
conversion of MAAM slows down, while EDMA is converted at
nearly the same rate as before. When approximately 70% of
EDMA is converted, the polymerization of MAAM accelerates
and proceeds at this higher rate practically until full conver-
sion. At this point both monomers are converted 100%.
These observations about the variations of the rate of con-
version for the individual monomers raise several questions,
which might, by the way, be asked for UV or thermal polymer-
ization as well. Is the polymer produced during the early stage
of polymerization the same as the polymer produced in the
middle or the late stage? Does the average composition of the
MIP or NIP reflect also its local compositions? If not, how do the
different compositions behave in binding? Does the polymer
produced during the later stages cover and shield the polymer
produced earlier? Our experiments of studying the template
202 a n a l y t i c a c h i m i c a a c t a 6 0 8 ( 2 0 0 8 ) 197–203
binding properties of the polymers produced with differ-
ent irradiation doses were meant to answer some of these
questions.
Before going on with the discussion, we must devote some
further attention to the polymerization rate measurements.
First of all, there is nothing surprising in observing differences
in the conversion rates of two co-monomers in a copolymer-
ization. This is a frequently observed phenomenon and has
been amply explained already by Flory [20]. It has also been
recognized in the MIP field [1], although it is rarely mentioned
in the MIP literature. A second, more serious problem with
the conversion rates is, that EDMA is a bifunctional monomer,
so it will disappear from solution when one of its two double
bonds reacts. The second double bond remains, however, usu-
ally intact when the first one binds the EDMA molecule to a
growing polymer chain. This pendant double bond is still avail-
able for further reaction, although restricted in its mobility
by hanging off a macromolecular chain. These pendant dou-
ble bonds are usually made responsible for the crosslinking of
the copolymer network but no other role is attributed to them.
Our results seem to show, that even after the disappearance
of EDMA from the solution there is still a lot of MAAM left in
the solution, ready to react with the pendant EDMA double
bonds. An alternative possibility for the late stage disappear-
ance of MAAM would be the homopolymerization of MAAM
to polyMAAM, which is insoluble in acetonitrile. Our binding
data, presented below, appear to rule out this second possibil-
ity, because the imprinting factor does not change appreciably
during the late stage of polymerization.
As we turn now to the template binding properties of the
MIPs and NIPs produced with different doses of radiation,
we still need to discuss the parameters used to character-
ize template binding. The MIP literature is quite confusing
on this point. One often finds data called “template bind-
ing capacity”, which seems to lack any reasonable definition,
because binding saturation of MIPs is rarely observed (mostly
due to solubility limitations, if nothing else). Chromatographic
characterization of MIPs is also lacking sound definitions and
experiments. We have shown earlier [21] – in agreement with
some other opinions [22] – that instead of such quantities, the
template binding isotherm of the MIP should be measured and
presented. In the present work we chose a simplified version
of isotherm measurement. One can calculate from any point
of a binding isotherm a distribution ratio, i.e., the ratio of the
equilibrium concentrations of the template on the polymer
and in the solution. Since MIP and NIP binding isotherms are
usually quite smooth, the distribution ratio does not change
too much in a narrow concentration range. Thus the distribu-
tion ratio is a suitable basis for comparisons, as long as the
equilibrium solution concentrations in its measurement are
similar. To realize this situation we have chosen a constant
phase ratio (6 ␮L solution per one mg of polymer). This was
important because at low conversion the amount of polymer
produced (from the same amount of starting material) was
obviously much less than at high conversion. In this way we
could keep the equilibrium solution concentrations within a
range of 0.60–0.89 mmol dm−3 .
Analysis of the phenytoin binding data (Figs. 4 and 5.)
shows several interesting features. The irradiation dose has
a great influence on the distribution ratio and a smaller but
significant effect on the imprinting factor. Both quantities
increase with the time of irradiation and reach a plateau
after 15 kGy dose. It is interesting that the distribution ratio
on both polymers shows a sharp increase between 10 and
15 kGy, where the solution phase contains almost no EDMA.
Since the binding sites must contain MAAM (our unpublished
experiments show that pure EDMA polymer does not bind
phenytoin appreciably), it is not surprising that phenytoin
binding increases when more MAAM is built into the polymer.
Surprisingly, however, the imprinting factor does not change
in this phase of polymerization, so that it is likely that even
during this final stage the polymer surface is similar to the ear-
lier formed polymer. This makes it likely that the new polymer
formed at the late stage contains not only MAAM but also the
pendant EDMA double bonds.
Finally, we have also observed that the distribution ratios
of the MIP and NIP after full conversion are somewhat higher
than the values we obtained earlier [23] with thermally poly-
merized phenytoin MIPs of the same composition, while the
imprinting factor is close to that of the thermally imprinted
MIP. The similarity of MIPs obtained by gamma and thermal
polymerization, respectively, may indicate that the observa-
tions made in this paper have some relevance also for the
other initiation methods.
4. Conclusions
In this paper we have investigated the process of gamma
polymerization of a MAAM-co-EDMA phenytoin MIP and its
control polymer (NIP). The polymers obtained after complete
conversion were found to behave similar to their thermally
polymerized counterparts. This result extends those of Sreeni-
vasan [24] to another kind of MIP.
We have followed the conversion of both monomers during
polymerization and measured the phenytoin binding prop-
erties of the MIPs and NIPs produced with increasing doses
of gamma irradiation. Several interesting observations have
been made.
The difference between imprinted and control polymers in
their binding properties has appeared already at quite early
stages of polymerization. This is surprising because these
early polymers show visible swelling (so that crosslinks may
have not yet fixed the binding site geometries).
The crosslinker EDMA has been consumed from solution
well before the functional monomer MAAM. This indicates
that many of the pending double bonds of the crosslinker
react only after all EDMA has been incorporated into the poly-
mer. These pending double bonds may react in two ways:
by making crosslinks among themselves or by binding the
remaining MAAM molecules from solution. Surprisingly, the
large number of binding sites formed during this late stage
of polymerization (between 10 and 15 kGy) have the same
imprinting factor as those formed earlier. This may allow the
conclusion that all binding sites are similar and thus all con-
tain MAAM bound to the pending double bonds, not in the
main chain of the polymer. Further on, the crosslinks formed
between 10 and 15 kGy dose by the many pending double
bonds do not improve the imprinting factor. This may be so
because crosslinking is less important for good binding sites
 a n a l y t i c a c h i m i c a a c t a 6 0 8 ( 2 0 0 8 ) 197–203
than thought before or because there occurs less crosslinking
during this late stage of polymerization purely by EDMA than
expected.
Some of these interesting observations may also be valid
for other types of polymerization (thermal and UV) but this
needs experimental verification. Further studies are also war-
ranted to investigate the useful properties of less crosslinked,
eventually swellable MIPs.
Acknowledgements
The authors are grateful to Zoltan Papp for gamma
irradiations; to the OTKA (T048912), the GVOP (3.2.2-2004-07-
0006/3.0) and to Institut de Recherches Internationales Servier
(I.R.I.S.), France for the financial support.
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1170295 Analytical followup of the gamma initiated synthesis of a molecularly imprinted polymer 7

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