Applied Surface Science 320 (2014) 138–145

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Applied Surface Science

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

Preparation of a magnetic molecularly imprinted polymer for

selective recognition of rhodamine B
Xiuying Liu, Dan Yu, Yingchao Yu, Shujuan Ji ∗
College of Food, Shenyang Agricultural University, No. 120 Dongling Road, Shenyang 110866, People’s Republic of China

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

Article history: A novel magnetic molecularly imprinted polymer (MMIP) was developed as an adsorbent to selec-
Received 22 February 2014 tively remove rhodamine B from real samples. The polymer was characterized by scanning electron
Received in revised form 30 July 2014 microscopy, Fourier-transform infrared spectroscopy, and thermo-gravimetric analysis. Static adsorp-
Accepted 21 August 2014
tion, kinetic adsorption, and selective recognition experiments were also performed to investigate the
Available online 28 August 2014
specific adsorption equilibrium, kinetics, and selective recognition ability of the MMIP. The MMIPs had
outstanding thermal stability, large adsorption capacity, and high competitive selectivity. When they
Keywords:
were used as dispersed solid-phase extraction adsorbents in real samples, rhodamine B recovery was
Rhodamine B
Magnetic molecularly imprinted polymer
79.97–81.88% and 75.56–79.74% in intra-day and inter-day reproducibility experiments with relative
Fluorospectrophotometry standard deviations lower than 2.62% and 4.28%, respectively. Extraction was optimized for yield and
efficiency. Precision, accuracy, and linear working range were determined under optimal experimen-
tal conditions. The limits of detection and quantification were 1.05 and 3.49 ␮g L−1 , respectively. These
results suggest MMIPs may be used for determination of rhodamine B in real samples.
© 2014 Elsevier B.V. All rights reserved.

1. Introduction may find it inaccessible because of the high cost of instrumentation

Rhodamine B (RhB) is a synthetic water-soluble dye with strong
fluorescence in solution. It is widely used for cell fluorescence stain-
ing, paper printing, textile dyeing, and in leather, cosmetics, and
pigment and paint industries [1]. RhB has also been introduced into
foodstuffs. Developed countries have forbidden the use of RhB as a
food colorant because of the risk to human health; however, devel-
oping countries such as Argentina do not regulate the use of RhB in
foods [2]. Though it is on the list of illegal food additives in China,
RhB contamination still occurs owing to its colorfastness and low
cost. RhB is harmful and could cause irritation to the skin, eyes, and
respiratory tract, if swallowed by humans or other animals. The
carcinogenicity, reproductive and developmental toxicity, neuro-
toxicity, and chronic toxicity of RhB in humans and animals have
been experimentally proven [3]. Owing to its harmful effects and
accompanying health risks, RhB monitoring in real samples has
become a concern.
Analytical methods for RhB dye determination involve chro-
matography [4,5], UV–visible spectrophotometry [6], and fluori-
metric determination [7]. Although chromatographic separation
enables more sensitive quantitative analysis, many laboratories
∗ Corresponding author. Tel.: +86 24 88498337; fax: +86 24 88487162.
E-mail address: jsjsyau@sina.com (S. Ji).
and qualified operators. Fluorospectrophotometry is an important
tool owing to its simplicity, low cost, and high sensitivity and
selectivity for fluorescent analytes. However, sample pretreatment
is a challenge because of low analyte concentrations and matrix
complexity. To reduce matrix interference and enrich the analyte,
pre-treatment processing is often necessary before instrumental
analysis [8].
Molecular imprinting technology is an attractive synthetic
method for creating specific binding sites in molecularly imprinted
polymers (MIPs) [9–11]. The field has expanded at an amazing
pace thanks to the significant advantages of molecularly imprinted
polymers such as predictable specific recognition, low cost, ease of
preparation, and high mechanical and chemical stability [12]. Sev-
eral polymer preparation methods have been developed such as
bulk polymerization [13–16], membrane polymerization [17,18],
precipitation polymerization [19–21], and surface-grafting poly-
merization [22–24]. Magnetic separation technology, in which
polymers are prepared by fabricating the MIP on the surface of a
magnetic substrate, has received considerable attention in recent
years for its potential application in extraction and separation
[25–27]. The resulting magnetic molecularly imprinted polymers
(MMIP) are added to a solution or suspension containing the target
analyte and stirred to allow the analyte to adsorb onto the magnetic
polymers. The polymers with captured analyte are then recovered
from the suspension with an appropriate magnetic separator. The

http://dx.doi.org/10.1016/j.apsusc.2014.08.122
0169-4332/© 2014 Elsevier B.V. All rights reserved.
 X. Liu et al. / Applied Surface Science 320 (2014) 138–145
analyte is eluted from the polymers and analyzed. This technology
provides a relatively rapid and easy way to remove magnetic poly-
mers from sample matrices by applying a magnetic field without
the need for tedious centrifugation or filtration [28,29].
We synthesized a novel MMIP by using Fe3 O4 magnetite as
the magnetically susceptible component and RhB as the template
molecule. The characteristics of the MMIP and its binding proper-
ties were investigated and the polymer was used as a sorbent for
extraction of RhB from real samples, followed by fluorospectropho-
tometry.
2. Materials and methods
2.1. Materials
Rhodamine B (≥99%), iron(II) chloride tetrahydrate
(FeCl2 ·4H2 O), iron(III) chloride hexahydrate (FeCl3 ·6H2 O), oleic
acid, polyvinylpyrrolidone (PVP), carmine, amaranth, tartrazine,
and sunset yellow were purchased (Aladdin-Reagent Co., Beijing,
China). Trihydroxymethylpropyl trimethylacrylate (TRIM) (Sigma-
Aldrich, USA) was used as received. Methacrylic acid (MAA) and
azobisisobutyronitrile (AIBN) were purchased from Sinopharm
Chemical Reagent Co., Ltd. (99%). All other reagents were at least
of analytical grade and were used without further purification.
2.2. Preparation of Fe3 O4 magnetite
Fe3 O4 magnetite was prepared by the co-precipitation method.
FeCl2 ·4H2 O (0.01 mol) and FeCl3 ·6H2 O (0.02 mol) were dissolved
in 100 mL water. The mixture was stirred vigorously at 80 ◦ C
under nitrogen gas, and then 40 mL sodium hydroxide solution
(2.0 mol L−1 ) was added to the clear yellow solution. After 1 h, the
black magnetic precipitates were isolated from the solvent with a
permanent magnet and washed repeatedly with deionized water
until the pH of the washings became neutral; the precipitate was
then dried at 60 ◦ C for 24 h.
2.3. Preparation of MMIPs
To prepare the MMIPs, the RhB (1.0 mmol) template was dis-
solved in 10 mL acetonitrile and 4.0 mmol of the MAA functional
monomer was added. The mixture was stirred for 1 h to prepare
the preassembly solution. The Fe3 O4 magnetite (1.0 g) was mixed
with 1.0 mL oleic acid and stirred for 5 min. Then 20 mmol TRIM and
the preassembly solution were added to the mixture of Fe3 O4 and
oleic acid. PVP (0.2 g), the dispersant, was dissolved in 50 mL of 80%
ethanol aqueous solution, and then added to the mixture, which
was dispersed by sonication for 1 h to prepare the prepolymer-
ization solution. AIBN (0.1 g) was added to the prepolymerization
solution, which was then stirred once again until fully homoge-
nized and purged with nitrogen for 5 min before being placed in
a water bath at 60 ◦ C for 24 h. Uniform magnetic microspherical
MIPs were obtained after screening through a 100-mesh sieve and
washed with a mixture of methanol and acetic acid (9:1, V/V) in
a soxhlet apparatus to remove the template and nonpolymerized
residue. Finally, the polymers were washed with water to reach
neutral conditions. The resultant MMIPs were dried for 12 h at 40 ◦ C.
The magnetic non-imprinted polymers (MNIPs) were prepared and
processed similarly but in the absence of RhB.
2.4. Binding experiment
The static adsorption experiment was performed by adding
50.0 mg MMIPs or MNIPs in centrifuge tubes containing 4.0 mL
RhB solution at various concentrations. The tubes were oscillated
for 12 h at room temperature to facilitate maximum RhB-polymer
139
binding, and then the MMIPs were isolated by an external magnetic
field.
The adsorption kinetics experiment was performed by detec-
ting the concentration of RhB in the solutions over time. The
concentration of RhB in the supernatant was measured with a flu-
orospectrophotometer. The amount of polymer-bound RhB was
calculated according to the following equation:
 
Ci − Cf × V × 1000
Q = (1)
m
where Q is the adsorption capacity of the polymer (␮g g−1 ), Ci is the
initial concentration of RhB (␮g mL−1 ), Cf is the final concentration
of RhB (␮g mL−1 ), V is the volume of the solution (mL), and m is the
mass of the dried polymer (mg). Each measurement was performed
in triplicate.
2.5. Selective recognition study
Selective recognition of the MMIPs was evaluated using several
colorants typically used in food processing. The experiment was
performed by adding 50.0 mg MMIPs or MNIPs to centrifuge tubes
containing 4 mL mixed colorants at 9 ␮g mL−1 . The tubes were
oscillated for 12 h at room temperature, and then the suspension
was separated by an external magnetic field and measured with a
UV–visible spectrophotometer. Each measurement was performed
in triplicate.
2.6. Extraction procedure
MMIPs were added to centrifuge tubes; the sample was added
and the mixture was oscillated. The MMIP and adsorbed RhB
were rapidly separated from the solution by an external magnetic
field. After the supernatant was discarded, RhB was eluted from
the MMIPs by using 3 × 3 mL methanol by oscillating during each
elution process. The eluate was measured by fluorospectrophotom-
etry.
2.7. Sample preparation
Wine samples were obtained from local markets. Initially, 1 mL
wine was diluted with 2 mL water. Then, 25 mg MMIP was added to
the solution and shaken at room temperature for 20 min. A magnet
was used to separate MMIP from the solution, followed by elu-
tion with 3 × 3 mL methanol. The supernatants were evaluated by
fluorospectrophotometry.
3. Results and discussion
3.1. Preparation of MMIPs
Synthesis of MIP-coated magnetic nanoparticles involved syn-
thesis of the magnetic Fe3 O4 nanoparticles, coating of the
nanoparticles with MIPs, removal of the template molecules, and
generation of the recognition sites. First, magnetic Fe3 O4 nanoparti-
cles were prepared by the co-precipitation method. Finally, an MIP
film was coated onto the surface of the nanoparticles. The entire
process is illustrated in Fig. 1. These MMIPs can be separated rapidly
in the presence of an external magnetic field (Fig. 2).
3.2. Characterization
The MMIPs were characterized by a series of physical tests. Scan-
ning electron microscope (SEM) images of the Fe3 O4 , MMIPs, and
MNIPs are shown in Fig. 3. The SEM image of Fe3 O4 showed that the
particle diameter was around 50 nm, with a regular spherical shape.
140 X. Liu et al. / Applied Surface Science 320 (2014) 138–145

Fig. 1. Schematic of the imprinting process.

Fig. 2. Separation process of MMIPs in the absence (a) and presence (b) of an external magnetic field.

Fig. 3. SEM images of the Fe3 O4 (a–c), MMIPs (d–f), and MNIPs (g–i) with magnification of 10,000 (a, d and g), 50,000 (b, e and h), and 200,000 (c, f and i).
 X. Liu et al. / Applied Surface Science 320 (2014) 138–145
Fig. 4. FT-IR spectra of the Fe3 O4 (a), and MMIPs (b).
SEM images of MMIPs and MNIPs showed that the surface of the
synthesized polymers was rough and porous, like a conglomeration
of beads with small cavities between larger ones. The differences
in surface characteristics suggest the Fe3 O4 cores were success-
fully coated with a polymer layer. FT-IR was performed to verify the
preparation of MMIPs (Fig. 4). A band at 566.40 cm−1 was attributed
to the vibration of Fe O Fe bonds in Fe3 O4 . The bands at 3434.18
and 1628.02 cm−1 indicated the presence of COOH on the surface
of Fe3 O4 (Fig. 4a). The intensity of Fe O Fe bonds was low (Fig. 4b).
The band at 3437.22 cm−1 indicated that the surfaces of both Fe3 O4
and MMIPs had many OH groups. A band at 2970.79 cm−1 was
attributed to C H2 stretching, while the band at 1467.14 cm−1 was
attributed to the bending vibration of C H from C H2 . Moreover,
many new absorption bands in the 1000.00–1800.00 cm−1 range
were attributed to the organic groups in MMIPs at different steps
of fabrication. MMIPs exhibited many absorption bands at 1735.39,
1262.29, and 1148.78 cm−1 , which were attributed to stretching
vibration of C O and to symmetric and asymmetric stretching
vibration of C O from TRIM, respectively. A band at 1389.20 cm−1
was attributed to the vibration of O . The low intensity of the
band at 566.40 cm−1 and the new bands in Fig. 4b confirmed that
MMIPs were successfully synthesized.
The XRD patterns of Fe3 O4 and MMIP were compared (Fig. 5).
There are several relatively strong diffraction peaks in the 2 range
of 10◦ –70◦ , quite similar to those of Fe3 O4 nanoparticles reported
by other groups [30]. The peak positions could be indexed to (2 2 0),
(3 1 1), (4 0 0), (4 2 2), (5 1 1) and (4 4 0) at 2 = 30.19◦ , 35.59◦ , 43.23◦ ,
53.57◦ , 56.99◦ , and 62.71◦ , respectively [JCPDS card (19-0629) for
Fe3 O4 ]. In addition, the XRD patterns of MMIP were similar to that
of Fe3 O4 ; thus, the coating procedure did not cause a structural
change in the Fe3 O4 .
The crystallite size was calculated from the Scherrer equation
[31]:
0.90
ε=
ˇ1/2 cos
where ε is crystallite size, and ˇ1/2 is the peak width (integral or
full width at half maximum) in radians. The crystallite size of Fe3 O4
powder prepared in this study was 32 nm, and the error was 4.42%.
In this study, Fe3 O4 , MMIP, and MNIP were heated from 0 ◦ C
to 880 ◦ C at 10 ◦ C min−1 under nitrogen at 20 mL min−1 . The
141
Fig. 5. X-ray diffraction (XRD) patterns of Fe3 O4 (a), and MMIPs (b).
thermo-gravimetric analysis (TGA) curves are given in Fig. 6. The
curves show that the weight loss of Fe3 O4 was 5.78% at 880 ◦ C,
demonstrating its thermal stability. The MMIP and MNIP curves
showed similar patterns, indicating the analogous thermal stabil-
ity of these polymers. The initial weight loss (0–330 ◦ C) could be
ascribed to the evaporation of water molecules. Both MMIP and
MNIP were almost stable at 330 ◦ C; however, the polymers decom-
posed gradually from 330 ◦ C to 450 ◦ C and more severely from
450 ◦ C to 500 ◦ C. The weight loss of MMIP and MNIP at 330 ◦ C,
450 ◦ C, and 500 ◦ C was 11.65%, 22.49%, and 77.55%, and 13.61%,
39.99%, and 88.50%, respectively. The slight difference in thermal
stability may be attributed to the difference in grafting density dur-
ing polymerization. Compared with MMIPs, Fe3 O4 showed little
weight loss even at 800 ◦ C. Thus, the remaining weight could be
attributed to Fe3 O4 stability. The results show that polymers were
grafted onto the surface of the Fe3 O4 nanoparticles; the grafting
yield of MMIP and MNIP was about 77.55% and 88.50%.
3.3. Adsorption capacity study
To study the adsorption capacity of MMIP, static adsorption and
adsorption kinetic experiments were performed. The isothermal
adsorption result is shown in Fig. 7. The molecular recognition
properties of MMIPs were evaluated by comparison with MNIPs.
We found that MMIPs have a stronger memory function and a
higher adsorption capacity for RhB than do MNIPs. With increasing
(2)
Fig. 6. Thermo-gravimetric analysis curves of Fe3 O4 (a), MMIPs (b), and MNIPs (c).
142 X. Liu et al. / Applied Surface Science 320 (2014) 138–145

400

350

300

250
MIP
200
N
150

100

50

0
carmine amaranth tartrazine sunset rhodamine
yellow B

Fig. 9. The adsorption selectivity of MMIPs and MNIPs for different analytes
(mMMIP = mMNIP = 0.05 g, V = 4 mL, t = 12 h, T = 20 ◦ C).

on MMIP and MNIP. The adsorption capacity increased rapidly in

the first 30 min and increased slightly before reaching equilibrium
Fig. 7. Adsorption isotherm of MMIP and MNIP (mMMIP = mMNIP = 0.05 g, V = 4 mL,
at 45 min. The RSD of MMIPs and MNIPs was less than 1.85% and
t = 12 h, T = 20 ◦ C).
1.86% at each time point (n = 3).

800
3.4. Selective recognition study
700
The specificity of the prepared MMIP was evaluated with several
600
colorants commonly used in food processing, including carmine,
500 amaranth, tartrazine, and sunset yellow. The experiment was per-
Q (μg g )
-1

MMIP formed by adding MMIPs or MNIPs to a solution containing the

400
MNIP mixed colorants and RhB. As shown in Fig. 9, the amount of RhB
300 bound to the MMIPs was higher than that bound to MNIPs. In
addition, the MMIPs exhibited a much larger binding capacity for
200
RhB than for other colorants, indicating that the MMIPs provided
100 high selectivity for RhB. The adsorption of other colorants by MNIP
0
and MMIP was similar, suggesting some non-specific adsorption
0 15 30 45 60 75 90 105 120 135 150 165 180 of other colorants by both polymers. This may be because some of
time (min)
these colorants have a smaller spatial configuration in comparison
to RhB. A few also have functional groups similar to RhB and may
Fig. 8. Adsorption kinetic curves of MMIP and MNIP (mMMIP = mMNIP = 0.05 g, thus account for non-specific adsorption.
V = 4 mL, c = 14 ␮g mL−1 , T = 20 ◦ C). The displacement of RhB bound by MMIPs in the presence of
competing compounds was calculated according to the following
concentrations of RhB, the adsorption capacity of MIP and NIP rose equation:
gradually and reached saturation at a higher concentration. (Qo − Q )
In addition, the equilibrium data were fitted to the Langmuir displacement (%) = × 100 (4)
Q
isotherm models. The form of the Langmuir model was expressed
by the following equations: where Qo and Q are the adsorption capacity of RhB by MMIPs in
the absence and presence of competing compounds. The displace-
KQm Ce ment of 2.12% showed that the specificity of the MMIPs for RhB
Q = (3)
1 + KCe would not be appreciably affected by competing compounds. The
where Q is the equilibrium adsorption capacity of the poly- MMIPs prepared in this study have great potential utility for the
mer (␮g g−1 ), K is the affinity constant, Qm is the maximum determination of RhB. The distribution coefficients (Kd ), selectivity
adsorption capacity (␮g g−1 ), and Ce is the equilibrium concen- coefficients (Ks ), and relative selectivity coefficients (Kr ) of carmine,
tration (␮g mL−1 ). We found that the equilibrium data were amaranth, tartrazine, and sunset yellow with respect to RhB can be
well represented by the Langmuir model. The fitting parame- obtained according the following equation:
ters are listed in Table 1. The maximum adsorption capacity of Q
Kd = (5)
MMIP (2350.52 ␮g g−1 ) was approximately 1.5 times that of MNIPs Cf
(1671.86 ␮g g−1 ). The RSD was less than 3.36% and 3.25% at each
Kd(RhB)
concentration of MMIPs and MNIPs (n = 3). Ks = (6)
We measured the adsorption kinetics from 5 min to 180 min. As Kd(X)
shown in Fig. 8, there were two phases for the adsorption of RhB Ks(MMIP)
Kr = (7)
Ks(MNIP)
Table 1
Langmuir fitting parameters for MMIP and MNIP. where X represents the competitive compounds and Ks(MMIP) and
−1 2
Ks(MNIP) are the selectivity coefficients of MMIPs and MNIPs. Val-
Qm (␮g g ) K R
ues of Kd , Ks , and Kr are summarized in Table 2. Values of Kr
MMIP 2350.52 0.031 0.956 showed the binding affinity of recognition sites for each analyte.
MNIP 1671.86 0.033 0.931
The values of Kr for carmine, amaranth, tartrazine, and sunset
 X. Liu et al. / Applied Surface Science 320 (2014) 138–145 143

Table 2
Adsorption selectivity of MMIPs and MNIPs.

Structure Compounds MMIP MNIP Kr

Kd (mL g−1 ) Ks Kd Ks

Carmine 14.11 8.33 14.42 3.94 2.12

Amaranth 29.33 4.01 29.92 1.90 2.11

Tartrazine 4.09 28.75 4.51 12.57 2.29

Sunset yellow 11.76 9.99 12.12 4.68 2.13

Rhodamine B 117.52 – 56.77 – –

yellow were 2.12, 2.11, 2.29, and 2.13, respectively, indicating During extraction, the MMIPs were dispersed in the sample
the descending order of recognition for competitive compounds solution to rebind analyte, and we investigated the minimum
was RhB > tartrazine > sunset yellow > carmine > amaranth. These amount of sorbent required for efficient recovery. Polymer amounts
results illustrated the success of the imprinting process. from 15 to 100 mg were applied to extract RhB from real samples.
The results indicated that 25 mg polymer was sufficient to yield
3.5. Optimization of extraction conditions 82.93% ± 2.53% recovery (Fig. 10c). Increasing the amount of MMIPs
yielded no improvement in RhB recovery.
In order to evaluate the applicability of MMIPs for separation and Elution times of 10 to 120 min were investigated to identify the
enrichment of RhB in real samples, we investigated the parameters optimum elution time to achieve thorough desorption of the ana-
affecting extraction performance, including solvent, time, amount lyte from the polymers. The results given in Fig. 10d show that
of MMIPs, elution time, and types of eluent. increasing elution time did not significantly improve recovery. A
Extraction solvent plays an important role in rebinding of the duration of 10 min appeared sufficient for complete desorption,
analyte because it determines the microenvironment of the binding with recovery of 81.49% ± 1.33%.
reaction and influences analyte stability [32]. In order to evalu- An appropriate eluent should be chosen to ensure complete elu-
ate the effect of extraction solvent, water, ethanol, acetonitrile, tion of analyte from the MMIPs. For this purpose, different types
acetone, and ethanol/water (50%, 80%, v/v) were investigated. The of solvent including methanol, acetonitrile, acetone, water, and
optimum results in Fig. 10a were obtained with water, yielding ethanol were applied. Methanol offered the highest recovery rates
a recovery rate of 79.65% ± 0.82%. Though water is not an ideal (Fig. 10e) and was selected as the eluent.
extraction solvent owing to the interference of formation of hydro-
gen bonds between the analyte and MMIP, our satisfactory results 3.6. Analytical performance
suggest polymer performance was not in fact influenced by water.
These findings are similar to those of previous reports [33,34]. Thus, The linearity of RhB in real samples was 0.04–1.4 ␮g mL−1 with
water was used as the sample extraction solvent during optimiza- a correlation coefficient of 0.9994. The regression equation was
tion.
Extraction time was also investigated. The adsorption of RhB
should achieve equilibrium to allow sufficient time to obtain sat- Table 3
isfactory recovery. We evaluated extraction times from 20 to The intra-day and inter-day precisions (n = 6).
120 min. The results given in Fig. 10b show a slight increase Spiked level Intra-day precision Inter-day precision
in recovery with increasing extraction time, although the differ- (␮g mL−1 )
Recovery (%) RSD (%) Recovery (%) RSD (%)
ences were not significant (P > 0.05). To reduce processing time,
we selected a 20-min extraction, which provided 82.09% ± 1.08% 0.528 81.88 2.62 79.74 4.28
2.160 79.97 1.97 75.56 3.24
recovery.
144 X. Liu et al. / Applied Surface Science 320 (2014) 138–145

(a) (b)
90 90
80 80
70
70
Recovery (%) 60
60

Recovery (%)
50
40 50
30
40
20
10
30
0 20
water 50% 80% ethanol acetonitrileacetone 10
ethanol ethanol
Types of extraction solvent
0
20 30 60 90 120
Adsorption time (min)

(c) (d)
90
100
80
90
70 80
Recovery (%)

60 70

Recovery (%)
50 60
40 50
30 40
20 30
20
10
10
0
0
15 25 50 75 100
10 30 60 90 120
Amounts of MMIPs (mg)
Elution time (min)

90 (e)
80
70
Recovery (%)

60
50
40
30
20
10
0
methanol acetonitrile acetone water ethanol
Types of eluent

Fig. 10. Optimization of extraction conditions.

Y = 0.3762X + 7.1601, where Y is absorbance and X is the concen- A comparison of this and previously published methods for RhB
tration of RhB (␮g mL−1 ). Limit of detection (LOD) and limit of determination revealed that the detection limit of our method is
quantification (LOQ), defined as the concentration corresponding superior (Table 4) [6,35,36]. The proposed method provides similar
to a signal equal to three and ten times the standard deviation of recovery and precision, and lower or similar LOD in comparison
the blank, were 1.05 and 3.49 ␮g L−1 , respectively. to prior methods. Our sample preparation process is convenient
Precision was evaluated by measuring relative standard devi- because the use of magnetic separation.
ations (RSDs) of intra- and inter-day tests. Intra-day precision
was determined by analyzing spiked wine samples (0.528 and 3.7. Reusability
2.160 ␮g mL−1 ) six times in one day. Inter-day precision was
determined over six days with similarly prepared spiked wine sam- Repeated binding/removal experiments were performed to test
ples. The results are shown in Table 3. RSDs of intra- and inter-day the stability and reusability of MMIPs. The recovery of RhB on
tests ranged from 1.97% to 2.62% and from 3.24% to 4.28%. The MMIPs decreased 8.98% after ten cycles, suggesting they can be
accuracy and precision of the method were acceptable. recycled.

Table 4
Characteristic performance of some recent studies on the determination of Rhodamine B.

Detection system Analyte Method LOD (␮g L −1 ) RSD References

UV–visible spectrophotometry RhB Cloud point extraction 1.3 2.40 [6]

HPLC-UV RhB Cloud point extraction 100–800 – [34]
UV–visible spectrophotometry RhB Solid phase extraction 3.14 5.0 [35]
Fluorospectrophotometer RhB Dispersive extraction 1.05 Present study
 X. Liu et al. / Applied Surface Science 320 (2014) 138–145
4. Conclusions
In this study, we developed a novel material based on the
use of magnetic Fe3 O4 nanoparticles coated with MMIP for pre-
concentration and determination of RhB in real samples prior
to spectrofluorimetric determination. Thermo-gravimetric analysis
indicated the polymers possessed good thermal stability (<330 ◦ C).
The polymer exhibited good recognition and selective ability, sug-
gesting its utility as an analytical tool. We developed a method to
detect RhB at low concentrations in wine samples by using MMIP
as enrichment sorbent coupled with fluorospectrophotometry. The
method provides the advantages of short analysis time and small
amounts of polymer, together with the inherent high sensitivity of
fluorescence spectroscopy to simplify analysis and enable detection
of trace amounts of RhB in real samples. The MMIP showed good
stability under optimized conditions and could be used for more
than 10 cycles with only a slight loss in specific adsorption behav-
ior. Although the application of MMIP in other sample types has not
yet been addressed, the quantitative recovery, accuracy, and pre-
cision demonstrated in this study indicate that the application of
MMIP opens a new window of opportunity for the determination
of RhB in food sample matrices.
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