Analytica Chimica Acta 569 (2006) 5865

A 9-vinyladenine-based molecularly imprinted polymeric membrane for the

efcient recognition of plant hormone
1
H-indole-3-acetic acid
Changbao Chen, Yanjun Chen, Jie Zhou

, Chunhui Wu
College of Chemistry and Material Science, Shandong Agricultural University, Taian, Shandong 271018, China
Received 1 December 2005; received in revised form 15 March 2006; accepted 15 March 2006
Available online 29 March 2006
Abstract
9-Vinyladenine was synthesized as a novel functional monomer for molecular imprinting techniques and its structure was established with
elemental analysis and
1
H NMR spectroscopy. The binding mechanism between this functional monomer 9-vinyladenine and the plant hormone
1
H-indole-3-acetic acid in acetonitrile was studied with UVvis spectrophotometry. Based on this study, using
1
H-indole-3-acetic acid as a template
molecule, a specic 9-vinyladenine-based molecularly imprinted polymeric membrane was prepared. Then, the resultant polymeric membrane
morphologies were visualized with scanning electron microscopy, and the membrane permselectivity for
1
H-indole-3-acetic acid,
1
H-indole-3-
butyric acid and kinetin was tested with separate experiments and competitive diffusion experiments. These results showed that the imprinted
polymeric membrane prepared with 9-vinyladenine exhibited higher transport selectivity for the template molecule
1
H-indole-3-acetic acid than
1
H-indole-3-butyric acid or kinetin. The membrane prepared with 9-vinyladenine also took on higher permselectivity for
1
H-indole-3-acetic acid
in comparison with the imprinted membrane made with methacrylic acid. It is predicted that the 9-vinyladenine-based molecularly imprinted
membrane may be applicable to the assay of
1
H-indole-3-acetic acid or for the preparation of a molecularly imprinted polymer sensor for the
analysis of
1
H-indole-3-acetic acid in plant samples.
2006 Elsevier B.V. All rights reserved.
Keywords: Molecular imprinting; Transport selectivity;
1
H-indole-3-acetic acid; 9-Vinyladenine
1. Introduction
Since being pioneered by Wulff [1,2], molecular imprinting
has become an effective way to prepare cross-linked polymer
materials that show a memory effect toward their templates,
and has been extensively studied by several groups [36]. As
a simple and elegant method for creating recognition sites for
template molecules in cross-linked polymers, it has gained much
attention in the eld of analytical chemistry for the past decade
[7,8]. The principle of this technique involves (i) the assembly of
a polymeric functional monomer around a template molecule
in a solution containing a high ratio of cross-linker, (ii) poly-
merization of the mixture by thermal initiation or irradiation
with UV light, and (iii) removal of the template molecule to
afford the imprinted polymer. Molecularly imprinted polymers
(MIPs) have been used in various applications, including station-
ary phases for chromatography [911], recognition of elements

Corresponding author. Tel.: +86 538 8242174; fax: +86 538 8242174.
E-mail address: zhoujie@sdau.edu.cn (J. Zhou).
in sensors [1214], mimics for biological receptors [3,15,16]
and solid phase extraction [1722]. Recently, the preparation,
morphology and diffusive permeability of MIP membranes have
aroused increasing attention [23,24]. The ability of MIP mem-
branes to change their diffusive permeability automatically by
responding to the presence of template molecules is the most
interesting phenomenon. MIP membranes may be applicable as
novel separation devices, chemical sensors with high stability
and selectivity, drug delivery systems (DDS) with molecular
recognition, and biomimetic membranes. However, it is desir-
able to increase the selectivity of these membranes to make them
suitable for practical application. In order to achieve this aimit is
necessary to improve the understanding of the basic nature and
recognition mechanism of MIP membranes by preparing MIP
membranes with the use of different functional monomers.
1
H-Indole-3-acetic acid (IAA) is a plant hormone that exists
in most plants and regulates growth and development in plants.
Takeuchi and co-workers [14,22] have prepared IAA-imprinted
polymer with N,N-dimethylaminoethyl methacrylate as the
functional monomer in the apolar solvent chloroform. The poly-
mer showed higher afnity for IAA in chloroform than in
0003-2670/$ see front matter 2006 Elsevier B.V. All rights reserved.
doi:10.1016/j.aca.2006.03.062
C. Chen et al. / Analytica Chimica Acta 569 (2006) 5865 59
a polar solvent, such as acetonitrile as a solid phase extrac-
tant [22] or sensor coatings on a quartz crystal microbalance
[14]. We attempted to prepare a IAA-imprinted membrane in
a polar solvent by using other functional monomer such as
9-vinyladenine. Methacrylic acid (MAA) is the most widely
used functional monomer in molecular imprinting. However,
because of the weak hydrogen-bonding interactions between
MAA and some template molecules in polar solvents, the MIPs
made with the use of MAA in polar solvents exhibited only
very weak recognition [22] and in some cases no recognition at
all [2527]. Therefore, it becomes necessary to use new func-
tional monomers for molecular imprinting applications in polar
solvents and also in aqueous media. In some previous works,
9-vinyladenine was used to synthesize poly(9-vinyladenine).
The polymer was immobilized on supports such as cellulose,
agarose and silica gel for the chromatographic separation of
nucleic acids in aqueous solutions by high-performance liquid
chromatography or afnity gel electrophoresis [28,29]. In the
present study, 9-vinyladenine (9-VA) was used as a novel func-
tional monomer for the preparation of a molecularly imprinted
polymer membrane for IAA in methanol and acetonitrile over
a cellulose support. In order to study the effect of functional
monomers on the recognition ability of imprinted membranes,
imprinting of IAA in a polar solvent using MAA as the func-
tional monomer was also performed. Selectivity of the resulting
imprinted membrane was compared with that of the 9-VA based
membrane.
2. Experimental
2.1. Materials and instruments
Adenine, ethylene carbonate, N,N-dimethylformamide
(DMF), 1,4-dioxane and thionyl chloride were presented by
J&K Chemical Tech. Ltd. Co.; DMF, thionyl chloride and
1,4-dioxane were puried before use. IAA,
1
H-indole-3-butyric
acid (IBA), kinetin (KT), ethylene glycol dimethacrylate
(EGDMA), MAA and Dowex 50wx4-200R were purchased
from Aldrich. EGDMA and MAA were distilled to remove the
inhibitor prior to use. 2,2-Azobiisobutyronitrile (AIBN) was
from Nankai University Special Reagent Factory. Acetonitrile
and methanol were of chromatographic grade. Other chemicals
were of analytical grade without further purication.
Vario III Elemental Analyzers (Germany Elementar com-
pany), an Sg-100c Comb Flash chromatograph (American Isco),
a Shimadzu UVvis 1601PC double-beam spectrophotometer,
a Bruker DPX300 FT-NMR machine, a Waters HPLC sys-
tem (a pump, Model 600; a manual injector, Model Rheodyne
7725i; ODS column, 150 mm3.9 mm i.d.; Model 2487 UV
absorbance detector) and a Hitachi model S-570 scanning elec-
tron microscope (SEM) were used.
2.2. Synthesis of the functional monomer 9-VA
2.2.1. Synthesis of 9-(2

-hydroxyehthyl) adenine
5.60 g (41.4 mmol) of adenine, 4.00 g (45.4 mmol) of ethy-
lene carbonate and a trace of sodium hydroxide were added
into 45 ml (581.5 mmol) of anhydrous DMF. The solution was
reuxed for 3.5 h and the solvent was removed under reduced
pressure. Recrystallization of the residue from ethanol yielded
9-(2

-hydroxyethyl) adenine as milky plates. Yield: 55%, mp:

238.3240.4

C (Literature [30] 238239

C). Anal. Calcd. for

C
7
H
9
N
5
O: C, 46.92; H, 5.06; N, 39.17. Found: C, 46.83; H,
5.061; N, 40.02.
1
H NMR (DMSO-d
6
): =8.15 ppm (s, 1H);
8.08 ppm (s, 1H); 7.23 ppm (s, 2H); 5.02 ppm (s, 1H); 4.18 ppm
(t, 2H, J =5.24 Hz); 3.75 ppm (t, 2H, J =5.24 Hz).
2.2.2. Synthesis of 9-(2

-chloroethyl) adenine
2.90 g (16.2 mmol) of 9-(2

-hydroxyehthyl) adenine and

28 ml (386 mmol) volume of distilled thionyl chloride were
heated and magnetically stirred in a parafn bath at 70

C for
2 h. After removing excess of thionyl chloride in vacuum, the
residue was dissolved in a 50 ml of water and treated with
25 ml of 5%of sodiumcarbonate solution, and the formed prod-
uct was ltered and dried. Recrystallization from ethanol gave
9-(2

-chloroethyl) adenine as colorless needles. Yield: 67.2%,

mp: 204.4205.6

C (Literature [30] 204205

C). Anal. Calcd

for C
7
H
8
N
5
Cl: C, 42.63; H, 4.06; N, 35.53. Found: C, 42.61;
H, 4.103; N, 36.09.
1
H NMR (DMSO-d
6
): =8.18 ppm (s,
1H); 8.16 ppm (s, 1H); 7.28 ppm (s, 2H); 4.51 ppm (t, 2H,
J =5.82 Hz); 4.08 ppm (t, 2H, J =5.82 Hz).
2.2.3. Synthesis of 9-VA
To a solution of 9-(2

-chloroethyl)adenine (0.84 g, 4.2 mmol)

in 69 ml (809 mmol) of 1,4-dioxane was added 5.5 ml of a
sodium methoxide solution newly prepared from methanol and
sodium. After stirring the solution for 40 h at room tempera-
ture, TLC showed that the starting material was no long visible
and a new product had appeared. Water was added until the
solution became clear, after which it was treated with Dowex
50wx4-200R(H
+
) until neutrality, and then evaporated to dry-
ness. The residue was puried with Flash Chromatograph using
chloroform/methanol (v/v =9/1) as an eluant and gave 9-VAas a
colorless solid. Yield: 74%, mp: 196.4197.6

C(Literature [30]
196197

C). Anal. Calcd for C

7
H
7
N
5
: C, 52.17; H, 4.38; N,
43.45. Found: C, 52.15; H, 4.383; N, 44.15.
1
H NMR (DMSO-
d
6
): =8.46 ppm (s, 1H); 8.17 ppm (s, 1H); 7.34 ppm (s, 2H);
7.28 ppm(dd, 1H, J
1
=16.02 Hz, J
2
=9.20 Hz); 6.08 ppm(d, 1H,
J =16.02 Hz); 5.11 ppm (d, 1H, J =9.20 Hz).
2.3. Spectrophotometric method
A series of solutions was prepared containing a xed con-
centration of IAA (1.0 10
5
M) and various amounts of 9-VA
in acetonitrile. The absorption spectra of these solutions were
determined with corresponding acetonitrile solutions of 9-VAas
references.
2.4. Preparation of IAA-imprinted polymeric membranes
AnIAA-imprintedpolymeric membrane with9-VAas a func-
tional monomer (P
9-VA-IAA
) was prepared according to the fol-
lowing procedure. A porous cellulosic membrane was soaked in
the prepolymerization mixture containing 3.4 mg (0.02 mmol)
60 C. Chen et al. / Analytica Chimica Acta 569 (2006) 5865
IAA, 32.1 mg (0.2 mmol) 9-VA, 0.38 ml (2 mmol) EGDMA,
3 mg AIBN, 2 ml methanol and 2 ml acetonitrile. The mixture
was degassed in an ultrasonic oscillator in ice-water bath for
10 min and stored in an icebox for 20 min. The cellulosic mem-
brane was then taken from the mixture, placed between two
glass plates and compressed until air bubbles were removed. It
was then polymerized with UV-irradiation at 365 nm for 10 h
at 04

C. After the glass plates were separated, the imprinted

membrane was washed with a 5% solution of sodium acetate in
methanol to wash out the template molecule IAA; then washed
with methanol. The membrane was kept in methanol at 4

C.
A reference non-imprinted membrane (P
NON
) was prepared
in the same way but in the absence of the template molecule and
worked up by the same procedure.
Another IAA-imprinted polymeric membrane (P
MAA-IAA
)
was prepared in similar manner by adding 42.4 l (0.5 mmol)
MAA instead of 32.1 mg (0.2 mmol) 9-VA as a functional
monomer.
2.5. Experiments of SEM
The surface morphology of the resultant polymer membranes
with the support of cellulosic membrane was observed with
a Hitachi model S-570 scanning electron micrograph (SEM).
The wet samples of the membranes were lyophilized. The small
circular disks (d 0.8 cm) from the center of the sample mem-
branes were obtained by fracturing the membranes at room
temperature. A thin layer of platinum was coated the small cir-
cular disk with an IB-5-sputter apparatus prior to observation.
2.6. Membrane transport experiments
The membranes were mounted between the two stirred cham-
bers inanH-shapedtwo-compartment cell withconstant stirring.
30 ml of a methanol solution, which was 1.0 10
5
M in a sub-
strate was placed in the feeding chamber, and pure methanol in
the receiving chamber. The amount of the transported analyte
was determined by spectrophotometry.
2.7. Examination of leakage of IAA in P
9-VA-IAA
According to the above procedure of transport experiments, a
control experiment was carried out with 30 ml of pure methanol
in the feeding chamber instead of a methanol solution of
1.0 10
5
M IAA. The concentration of IAA in the receiving
chamber as a function of permeation time was detected with
spectrophotometry.
2.8. Evaluation of membrane separation performance
The polymeric membranes were placed between the same
devices with the above transport experiments. A methanol solu-
tion consisting of 5.0 10
5
Mconcentrations of IAA, IBAand
KT was in the feeding cell, and pure methanol in the receiving
cell. The amount of each transported analyte was quantied by
HPLC with an eluent of methanol/0.6% acetic acid/acetonitrile
Fig. 1. Synthetic scheme of 9-vinyladenine.
(47/48/5, v/v/v) at a ow rate of 1.0 ml/min and the detection
wavelength 254 nm.
3. Results and discussion
3.1. Synthesis of 9-VA
Adenine is a nucleotide base and its important function is the
control of genetic processes by hydrogen bonding interaction
with other corresponding nucleotide bases. The hydrogen bond-
ing interaction of adenine directed us to use 9-VAas a functional
monomer in molecular imprinting. So we synthesized 9-VA
according to Uedas method [31] as shown in Fig. 1, with slight
modication. Adenine was rst converted to the correspond-
ing9-hydroxyethyl compound, the hydroxyethyl compoundthus
obtained was chlorinated to afford its derivative, which was fur-
ther dehydrochlorinated to give the nal compound.
3.2. Spectrophotometric analysis
In order to establish the hydrogen bonding and electrostatic
interaction between IAA and 9-VA in the prepolymerization
Fig. 2. Absorption spectra for IAA and 9-VA system in acetonitrile:
a
0
=1.0 10
5
M; b
0
(10
4
M): 00.00, 10.2, 20.4, 30.6, 40.8, 51.0;
l =1 cm, with corresponding 9-VA acetonitrile solutions as references.
C. Chen et al. / Analytica Chimica Acta 569 (2006) 5865 61
mixture, we investigated the stability of the cooperative
hydrogen-bonded and ionically interacted complex by spec-
trophotometry. A series of solutions was prepared, each with
a xed concentration of IAA (1.0 10
5
M) in acetonitrile
and a varying, increasing amount of 9-VA. Absorption spectra
of these solutions were determined with the corresponding
acetonitrile solutions of 9-VA as references (Fig. 2).
The absorption spectra show a small red shift of the absorp-
tion peaks with the increase of 9-VA. This indicated that a
complex had been formed between IAA and 9-VA from elec-
trostatic and/or hydrogen bonding interaction. The association
constant of the complex was determined according to our earlier
method [32] with slight modication. For an analytical concen-
tration of 9-VA (b
0
) greater than that of IAA (a
0
), the binding
reaction can then be expressed as
A +nB = C (1)
where, A and B stand for the reactants IAA and 9-VA, C repre-
sents the complex formed between them and n is stoichiometric
coefcient of B. In the above equilibrium, the association con-
stant is expressed as
K =
[C]
[A][B]
n
(2)
From the equilibrium of materials, yield:
[A] +[C] = a
0
(3)
[B] +n[C] = b
0
(4)
Fig. 3. Plot of A/b
n
0
vs. A at 258 nm. Curves: (1) n =1, ordinate: A/b
0
(10
2
M
1
); (2) n =2, ordinate: A/b
2
0
(10
6
M
2
); (3) n =3, ordinate: A/b
3
0
(10
11
M
3
).
Because b
0
a
0
, then [C] of the above Eq. (4) can be ignored,
that is,
[B] b
0
(5)
By substituting Eqs. (3) and (5) into Eq. (2), we obtain:
[C] =
a
0
b
n
0
K
1 +b
n
0
K
(6)
The absorbance of the solution is
A =
A
l[A] +
B
l[B] +
C
l[C]
=
A
la
0
+
B
lb
0
+(
C

A
n
B
)l[C] (7)
Fig. 4. Schematic illustration of the molecular imprinting procedures in this paper.
62 C. Chen et al. / Analytica Chimica Acta 569 (2006) 5865
We dene A
0,A
=
A
la
0
, A
0,B
=
B
lb
0
, then:
A = A A
0,A
A
0,B
= (
C

A
n
B
)l[C] = l[C]
(8)
Substituting Eq. (6) into Eq. (8) yields:
A
b
n
0
= KA +Kla
0
(9)
where, A
0,A
and A
0,B
can be evaluated from pure acetonitrile
solution of IAA and 9-VA. n =1, 2, 3, . . ., i.e. n is a simple
positive integer. When n =1, 2, 3, the plots of A/b
n
0
versus
A are shown in Fig. 3. Fig. 3 illustrates that when n =1 or 3,
the plots of A/b
n
0
versus A are curves. While n =2, the plot
of A/b
n
0
versus A is linear. In this situation, K is calculated
to be 1.597 10
9
M
2
. It shows that 1:2 complexes are formed
and have considerable stability. After cross-linker and initiator
were added in the solution, the complexes were imprinted in the
polymeric matrix according to the scheme shown in Fig. 4.
3.3. Characteristics of SEM to P
9-VA-IAA
and P
NON
When a cellulosic membrane was soaked into the prepoly-
meration solution, some self-assembly complexes were loaded
into the pores of the cellulosic membrane material. After poly-
merization and removal of the template molecules, the imprinted
polymeric membrane attached to the support material was
smooth, translucent, and exible by direct observation [33].
There was nodifference betweenP
9-VA-IAA
andP
NON
inmacrog-
raphy. However, changes of porous structure in the membranes
were detected with scanning electron microscopy (Fig. 5). As
indicated by offwhite marks in moderate magnication images
of the imprinted and non-imprinted polymeric membranes, there
were some asymmetric large pores in P
9-VA-IAA
, while some
small and symmetric pores existed in the P
NON
. These macrop-
ores in P
9-VA-IAA
are likely to be produced by the participation
and removal of the template molecules (IAA) in the polymer for-
mation of P
9-VA-IAA
. In addition, better densely packed agglom-
erates of P
NON
could be distinguished from those of P
9-VA-IAA
.
The asymmetric macro porous structure in P
9-VA-IAA
seems to
have a decisive impact on P
9-VA-IAA
transport selectivity.
3.4. Transport selectivity of polymeric membranes
In order to understand the complementary relationship
between the shape of the template molecules and the functional
groups in the cavities of P
9-VA-IAA
, the permselectivity of the
imprinted membrane P
9-VA-IAA
was assessed by diffusion stud-
ies of the substrates IAA, IBA and KT, which are plant growth
regulators (Fig. 6). Fig. 7 shows diffusion uxes of the substrates
(in separate experiments) across P
9-VA-IAA
(a) or P
NON
(b).
Diffusion rates of the substrates across the non-imprinted con-
trol membrane were slow. However, IAA diffused through the
imprinted membrane at a higher rate than IBA or KT. The most
signicant difference in diffusion rates through P
9-VA-IAA
is that
between IAA and KT. This indicates that the imprinted mem-
brane is permselective to the template molecule IAA. It should
Fig. 5. Scanning electron micrograph of P
9-VA-IAA
(A) and P
NON
(B).
be noted that IBA molecules were transported faster through the
imprinted membrane than KT molecules probably because the
molecular structure of IBA resembles to that of IAA, while the
molecular structure of KTis evidently different fromthat of IAA
(Fig. 6). The distinct different transport rates of the imprinted
and non-imprinted membranes indicated that the imprinting pro-
cess created channels through which template molecules could
pass successfully. In this way, a kind of continuous and fast
transport process was accomplished based on selective shape
and size and position of functional groups in the channels that
could recognize the best suitable template molecule. In con-
trast, the non-imprinted control membrane P
NON
did not show
any imprinted effect and the dense and small pores are arranged
in a disorderly manner that cannot generate channels. It is also
possible that the routeways in the P
NON
were jammed so that
the substrate molecules were transported at a low rate [34].
The different diffused behaviour of substrates in the molecu-
larly imprinted and non-imprinted control membranes results
from imprinting effects. Imprinting creates a specic micropore
fraction for template molecules and the connectivity of pores,
C. Chen et al. / Analytica Chimica Acta 569 (2006) 5865 63
Fig. 6. Molecular structures of substrates.
which are not present in the non-imprinted membranes. Table 1
shows the selectivity (ratios of diffusive ux of IAA to that of
the other substrates) of the imprinted and non-imprinted mem-
branes. This also shows that the imprinted membrane has very
high permselectivity for its template molecule, IAA.
In order to understand the reason that IAA is allowed to
pass through the imprinted membrane P
9-VA-IAA
at a greater
rate while the template molecule IAA was completely removed
from the imprinted membrane P
9-VA-IAA
in an elution process,
we substituted methanol for substrates and carried out the con-
Table 1
Selective factors of tested substrates for P
9-VA-IAA
and P
NON
in separate
experiments
Polymeric membranes Selective factors
IAA/IAA IAA/IBA IAA/KT
P
9-VA-IAA
1.00 1.55 2.03
P
NON
1.00 0.86 1.10
Solvent: methanol; [Initial substrate] =1.0 10
5
M; permeation time: 14 h;
effective volume of diffusion cell: 30 ml; effectual area of membranes: 3.14 cm
2
.
trol experiment under similar diffusion conditions. Curve 0 in
Fig. 7(a) shows the IAA concentration in the receiving cham-
ber is close to zero during whole permeation process. This
indicates that there was no leakage of IAA from P
9-VA-IAA.
In
addition, the concentration of IAA in the feed chamber was also
determined when IAA diffused across the imprinted membrane
P
9-VA-IAA
(curve 4 in Fig. 7(a)). The sum of IAA concentra-
tions in the receiving and feed chambers at each permeation
interval is almost equal to the initial IAA concentration in the
feed chamber (curve 5 in Fig. 7(a)). This further indicates that
there are recognition sites in P
9-VA-IAA
for the template molecule
IAA and the IAA leakage from P
9-VA-IAA
is negligible. Theo-
retically, the total IAA in the two chambers should be less than
the amounts of initial IAA because of binding of IAA to the
imprinted membrane. In effect, the amount bound to P
9-VA-IAA
is not appreciable in comparison with that contained in the solu-
tion. This is because there is very small polymer fraction in the
very thin and imprinted composite membrane, the effectual dif-
fusion area of which is 3.14 cm
2
, while a large amount of IAA
is permeated in the long permeation process.
3.5. Evaluation of membrane separation capacity
In order to further understand the selectivity of the imprinted
membrane P
9-VA-IAA
, we undertook competitive transport exper-
iments with P
9-VA-IAA
, where the three substrates IAA, IBA and
KT were applied simultaneously. The concentrations of the sub-
strates were determined by HPLC and the selective factors were
calculated. Values of the selection factors obtained are shown in
Fig. 7. Timetransport curves of testedsubstrates throughP
9-VA-IAA
(a) or P
NON
(b). Initial concentrationof substrates infeedingchamber: 1.0 10
5
M; measurement
wavelength: 258 nm; (10
4
l mol
1
cm
1
): IAA-4.24, IBA-4.72,KT-4.50; Curves: (0) leakage of IAA, (1) 1

-KT in receiving chamber, (2) 2

-IBA in receiving
chamber, (3) 3

-IAA in the receiving cell, (4) IAA in the feed chamber, (5) gross IAA of the feeding and receiving chamber.
64 C. Chen et al. / Analytica Chimica Acta 569 (2006) 5865
Table 2
Selective factors for tested substrates in P
9-VA-IAA
and P
MAA-IAA
in competitive
diffusion experiments
Permeation
time (h)
Selective factors
P9-VA-IAA PMAA-IAA
IAA/IAA IAA/IBA IAA/KT IAA/IBA IAA/KT
2 1.00 2.00 2.37 1.20 1.56
4 1.00 1.92 2.54 1.11 1.15
8 1.00 1.59 1.99 1.13 1.21
14 1.00 1.38 1.67 1.13 1.19
Solvent: methanol; initial solution: a solution containing 5.0 10
5
M of IAA,
5.0 10
5
M of IBA and 5.0 10
5
M of KT; effective volume of diffusion
chamber: 30 ml; effectual area of membranes: 3.14 cm
2
.
Table 2. Table 2 indicates that IAA was transported at a greater
rate than IBA and KT, but that selectivity was reduced with
permeation time. This is due to the decrease in the differences
of substrate concentrations in the feed and receiving chambers.
IAA still passed smoothly, continuously and quickly through
the polymeric membranes with the driving force of preferential
and reversible binding and exchange between it and MIP sites
in the membrane, although other concomitant molecules could
disturb its permeation. The existence of the disturbing effect
slightly reduces the permselectivity of the polymeric membrane
for the template molecule IAA as compared to that of the sepa-
rate experiments.
Table 2 also suggests that the permselectivity of the P
9-VA-IAA
for IAA is higher than that of the P
MAA-IAA
by comparing
their selective factors. The selective factors of P
9-VA-IAA
for
IAA/IBA and IAA/KT are 1.38 and 1.67, while those of the
P
MAA-IAA
are 1.13 and 1.19, respectively after a permeation time
of 14 h. The results agree with that of Kugimiya and Takeuchi
[22] and further demonstrate that MAA is not a good functional
monomer for molecular imprinting in a polar solvent. It is pre-
dicted that P
9-VA-IAA
can be used as a separation membrane in
IAA enrichment and decontamination for some plant samples
or as sensor coatings on a quartz crystal microbalance in IAA
detection in some plant materials. However, further experiments
will be carried out for real application of the imprinted mem-
brane P
9-VA-IAA
.
The selective transport arises from a process that involves
reversible complexation and exchange between IAA and the
imprinted sites in the membrane by cooperative hydrogen bond-
ing and electrostatic bonding interaction. Facilitated transport
of IAA across the IAA-imprinted polymeric membrane com-
pared to IBA or KT is attributable to the complementary shapes
and functional groups between the IAA and the imprinted sites.
Concentration differences serve as driving force for molecu-
lar transport through the membranes. Substrate molecules that
bonded to the membrane could either dissociate into the receiv-
ing chamber or back into the feed chamber. Because of the
concentration gradient, most of the substrate molecules diffused
to the receiving chamber [35]. The size of IBA and the shape of
KT were not complementary to the imprinted sites of P
9-VA-IAA
,
so there were weak complexation and exchange between IBA
or KT and the imprinted sites in the membrane. This resulted in
low transport rate with the minor selectivity when IBA or KT
was through P
9-VA-IAA
. Further experiments and a more inten-
sive evaluation of the system will be necessary to understand
better the recognition of 9-VA-based imprinted polymer.
4. Conclusions
9-VA was synthesized and exploited as a novel functional
monomer in molecular imprinting. After certifying the feasibil-
ity of imprinting between IAA and 9-VA by spectrophotomet-
ric analysis, the imprinted polymeric membrane was prepared.
The diffusion experiments of substrates showed that the IAA-
imprinted polymeric membrane exhibited high transport selec-
tivity for IAA in comparison with IBA and KT. By comparing
the permselectivity of P
9-VA-IAA
and P
MAA-IAA
, we found that 9-
VA-based polymeric membrane showed higher permselectivity
for IAA than MAA-based membrane in a polar solvent. Molec-
ular recognition of P
9-VA-IAA
is on the basis of the molecularly
imprinting effects of IAA. The characteristic seems to be appli-
cable to the assay of IAA or to be a feasible approach to prepare
an IAA-imprinted polymer sensor for the determination of IAA
in plant samples.
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