Sensors and Actuators B 228 (2016) 767773

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Sensors and Actuators B: Chemical

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

A turn-on uorescent probe for Hg2+ detection by using gold

nanoparticle-based hybrid microgels
Yecang Tang , Yi Ding, Ting Wu, Liying Lv, Zhicheng Yan
College of Chemistry and Materials Science, Anhui Normal University, The Key Laboratory of Functional Molecular Solids, Ministry of Education, Anhui
Laboratory of Molecule-Based Materials, Wuhu 241000, China

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

Article history: Hybrid microgels with Au nanoparticles (AuNPs) immobilized in poly(N-isopropylacrylamide-

Received 15 October 2015 co-2-(dimethylamino)ethylmethacrylate) (P(NIPAM-co-DMA)) microgels have been prepared. The
Received in revised form 20 January 2016 as-synthesized hybrid microgels showed excellent thermo- and pH-responsive properties and high sta-
Accepted 23 January 2016
bilities. 1-pyrenebutyric acid (PBA) molecules were then adsorbed on the AuNP surfaces via electrostatic
Available online 27 January 2016
interactions to form PBA-AuNPs/P(NIPAM-co-DMA) composites that were used as probes for the detec-
tion of Hg2+ . The uorescence of PBA was effectively quenched through uorescence resonance energy
Keywords:
transfer between AuNPs and PBA molecules. However, the addition of Hg2+ ions caused the uorescence
Fluorescence turn-on
Mercury detection
turn-on dramatically and 16 times uorescence enhancement was obtained, which is attributed the fact
Hybrid microgels that Hg2+ ions replace the adsorbed PBA molecules. Under the optimal conditions, the uorescence inten-
Gold nanoparticle sity was proportional to the concentration of Hg2+ ions in the range of 0.161.60 M with a detection
limit of 31 nM. Meanwhile, some other metal ions did not interfere with the detection of Hg2+ , suggesting
a good selectivity for Hg2+ ions sensing.
2016 Elsevier B.V. All rights reserved.

1. Introduction which exhibits uorescence enhancement in the presence of Hg2+

Mercury, a well-known highly toxic heavy metal, possesses
serious threats to human health and the environment. Long-term
exposure to mercury, even at very low level concentrations, can
cause serious and permanent damage to the central nervous and
endocrine system [13]. One of the most common and stable
forms of mercury pollution is solvated Hg2+ ions. Therefore, the
development of highly sensitive and selective methods for detect-
ing Hg2+ ions is of great signicance. To achieve that, various
methods are currently available for the measurement of Hg2+ ,
such as atomic absorption/emission spectroscopy [4,5], inductively
coupled plasma mass spectrometry [6], electrochemistry [79],
colorimetry [2,10,11], uorescence spectroscopy [1222] and so
on. Among them, the uorescence spectroscopy offers signicant
advantages over other detection methods because of its high sensi-
tivity, simplicity, inexpensiveness, and in situ detection capability
[23].
To date, a number of uorescent probes for Hg2+ ions have been
reported and can be categorized into two types by uorescence
enhancement or quenching. The rst type is the turn-on probe,
Corresponding author. Fax: +86 5533869303.
E-mail address: tycang@mail.ahnu.edu.cn (Y. Tang).
ions [2327]. The second type is the turn-off probe, based on the
uorescence quenching of the uorophores upon Hg2+ complexa-
tion [16,19,20,28]. The turn-on probe is more preferable than the
turn-off probe because the latter can produce false positive sig-
nals caused by other quenchers in practical samples [25,26]. Thus,
it would be advantageous to design turn-on uorescent probes to
detect Hg2+ ions.
Gold nanoparticles (AuNPs) have an extremely high extinc-
tion coefcient and a broad absorption spectrum, which allow
them to be employed as exquisite quenchers of uorescent dyes
through energy and/or electron transfer process. AuNP-based u-
orescence assays are emerging as alternative approaches for metal
ions, small molecules and DNA detection, providing high sensitivity
and specicity [28,29]. For example, Chang et al. [27] have reported
a rhodamine B (RB)-AuNPs system for the sensing of Hg2+ ions.
The uorescence of RB adsorbed onto AuNP surfaces is strongly
quenched. The presence of Hg2+ ions causes the RB molecules to
be detached from the AuNPs, leading to a large turn-on uores-
cence enhancement. Its selectivity can be improved by modifying
the AuNP surfaces with thiol ligands and adding a chelating ligand
to the sample solutions. However, AuNP-based probes are gener-
ally unstable and easily aggregated in real samples owing to their
high surface energy, which limits their practical application. To
overcome this limitation, bovine serum albumin has been used to

http://dx.doi.org/10.1016/j.snb.2016.01.112
0925-4005/ 2016 Elsevier B.V. All rights reserved.
768 Y. Tang et al. / Sensors and Actuators B 228 (2016) 767773

Table 1
Comparation of different probes for the determination of Hg2+ ion.

Methods Probe Linear range Detection limit Ref.

Colorimetric l-tyrosineAuNPs 33300 nM 53 nM [46]

Colorimetric CitrateAuNPs 11000 nM 2.9 nM [2]
Colorimetric CTABstabilized anisotropic nanogolds 010,000 nM 122 nM [10]
SERS Magnetic silica sphere @Au core/shell 0.11000 nM 0.1 nM [47]
SERS BismuthiolIIAuNPs 0150 nM 30 nM [48]
Fluorescence PEGAuNPs 5100 nM 2.24 nM [49]
Fluorescence H2 O2 amplex ultraRedAuPs 51000 nM 4.0 nM [50]
Fluorescence RBPEGAuNPs 1050 nM 2.3 nM [32]
Fluorescence Conjugated polymerbased lm sensor 20030,000 nM 100 nM [51]
Fluorescence LBL on polymer particles 200080,000 M 200 nM [17]
Fluorescence Composite turnon 404000 nM 32 nM [24]
Fluorescence Au nanodotsPNIPAM 220 nM 1.9 nM [36]
Fluorescence PBA-AuNPs/P(NIPAMcoDMA) 1601600 nM 31 nM This work

Abbreviations: surface-enhanced Raman scattering (SERS), hexadecyl trimethyl ammonium bromide (CTAB), polyethylene glycol (PEG), Rhodamine B (RB).

stabilize rhodamine 6G/AuNPs probe that could sense mercury ions
under high salt solutions [30,31]. By employing similar principle,
Hg2+ probe based on rhodamine B isothiocyanate-poly(ethylene
glycol)-comodied AuNPs that can be well-dispersed in various
complex solutions has been designed [32].
Microgels have been used as templates for the in situ synthesis of
inorganic nanoparticles. A major advantage arises from its polymer
networks, which can effectively hinder nanoparticles aggregation
and thus enhance their chemical stability. In particular, the porosity
affords an opportunity for a rapid diffusion of analytes into and out
of the networks [33]. Thus, hybrid microgels combining inorganic
nanoparticles and organic polymer networks have attracted sig-
nicant interest in the last decade. Their unique properties have
been utilized in organic catalysis [34] and metal-enhanced u-
orescence [35]. Recently, poly(N-isopropylacrylamide) (PNIPAM)
microgels incorporated with gold nanodots have emerged as a
selective and sensitive probe for the detection of mercury ions
through Hg2+ -induced photoluminescence quenching of gold nan-
odots. More importantly, these hybrid microgels can be easily
puried by a simple centrifugation and display great stability
against salt [36]. Nevertheless, few studies have concentrated on
using hybrid microgel-based turn-on uorescent probes for Hg2+
ions detection.
In this work, we synthesized hybrid microgels with
AuNPs immobilized in the thermo-and pH-responsive poly(N-
isopropylacrylamide-co-2-(dimethylamino) ethylmethacrylate)
(P(NIPAM-co-DMA)) microgels. This hybrid microgel was devel-
oped as a selective turn-on uorescent probe for the detection
of Hg2+ ions. As shown in Scheme 1, negatively charged 1-
pyrenebutyric acid (PBA) dyes were rst adsorbed into positively
charged AuNPs/P(NIPAM-co-DMA) hybrid microgels, which
resulted in the uorescence quenching of PBA. However, in the
presence of Hg2+ , the uorescence of PBA switched to turn-on.
2. Experimental
2.1. Materials
N-isopropylacrylamide (NIPAM, 98%), 2-
(dimethylamino)ethylmethacrylate (DMA, 99%) and
1-pyrenebutyric acid (PBA) were purchased from Aladdin Chem-
ical Co., Ltd. NIPAM was puried by recrystallization from a
benzene/n-hexane mixture. DMA was distilled under reduced
pressure. N,N -methylenebisacrylamide (MBA, 98%), ammonium
persulfate (APS, AR), sodium dodecylsulfate (SDS, AR), chloroau-
ric acid hydrated (HAuCl4 4H2 O, AR), and sodium borohydride
(NaBH4 , 96%) were obtained from Sinopharm Chemical Co., Ltd.
MBA was puried by recrystallization from methanol and APS was
recrystallized from the mixed solvent of ethanol and deionized
water. Other chemical reagents were of analytical grade and used
as received. The buffer solution was freshly prepared with KH2 PO4 ,
Na2 HPO4 12H2 O, and Na3 PO4 12H2 O. Deionized water was used
throughout the experiments. P(NIPAM-co-DMA) microgels with
the molar ratio of 4:1 for NIPAM to DMA were prepared by a
dispersion polymerization method reported previously [37].
2.2. Characterization
The morphologies of the hybrid microgels were observed by
using a Tecnai G20 transmission electron microscope (TEM) with
an accelerating voltage of 200 kV. Few drops of dilute aque-
ous dispersion were dropped onto a carbon-coated copper grid.
The average hydrodynamic radius (Rh ) was measured by an
ALV/DLS/SLS-5022F spectrometer with a multi- digital time cor-
relation (ALV5000). A cylindrical 22 mW UNIPHASE He-Ne laser
(0 = 632 nm) was used as the light source. Each sample was l-
tered through a 0.8 m nylon lter to remove dust. All dynamic
laser light scattering (DLS) measurements were performed at a
scattering angle of 90 . Thermogravimetric analysis (TGA) was con-
ducted on an SDT Q600 under argon atmosphere. The samples were
heated from 20 to 700 C at a rate of 10 C min1 . UVvis absorption
spectra were measured on a Hitachi U-4100 spectrophotometer.
Fluorescence spectra were obtained by using a Hitachi F-4500 spec-
trouorometer equipped with an R3896 red-sensitive multiplier
and 1 cm quartz cuvette.
2.3. Synthesis of AuNPs/P(NIPAM-co-DMA) hybrid microgels
25 mL of P(NIPAM-co-DMA) microgels (2.8 mg mL1 ) and 10 mL
of aqueous HAuCl4 solution (1 mg mL1 ) were mixed and stirred
overnight at room temperature under nitrogen atmosphere. The
mixture was dialyzed against deionized water for 2 h to remove
unbound AuCl4 ions. Subsequently, the dispersion was cooled
in an ice bath and 1 mL of freshly prepared NaBH4 solution
(0.5 mg mL1 ) was slowly added. The reduction reaction was
allowed to proceed for 12 h. The resultant hybrid microgels were
further puried by dialyzing against deionized water and stored in
a refrigerator at 4 C with a concentration of 0.5 mg mL1 .
2.4. Fluorescence quenching of PBA by the hybrid microgels
A stock solution of PBA (10 M) was prepared with deionized
water. 250 L of PBA and different aliquots of hybrid microgels
(0.5 mg mL1 ) were mixed in a series of colorimetric tubes and
then diluted to 1.25 mL. After equilibration at room temperature
for 30 min, the uorescence spectra of the mixtures were recorded
with an excitation wavelength of 346 nm. The slit widths of excita-
tion and emission were 10 and 5 nm, respectively.
 Y. Tang et al. / Sensors and Actuators B 228 (2016) 767773 769

Scheme 1. Schematic illustration of the uorescent sensing of Hg2+ ions with PBA-AuNPs/P(NIPAM-co-DMA) composites.

2.5. Procedures for Hg2+ determination

250 L of PBA (10 M) and 500 L of hybrid microgels

(0.5 mg mL1 ) were mixed and then various amounts of Hg2+ stock
solution (20 M) were added. The nal volume was adjusted to
1.25 mL with PBS buffer (10 mM, pH 6.0) and the mixture was
equilibrated for 30 min before the spectral measurement. The u-
orescence spectra were measured under the same experimental
conditions as described above. In order to examine the selectivity of
this probe toward Hg2+ ions, various relevant metallic ions includ-
ing Co2+ , Ba2+ , Pb2+ , Cu2+ , Fe2+ , Fe3+ , Al3+ , Mg2+ , Ca2+ , Zn2+ , Na+ , Ag+
and K+ (12.8 M) were analyzed in the absence and presence of
1.28 M Hg2+ .

3. Results and discussion

3.1. Preparation of AuNPs/P(NIPAM-co-DMA) hybrid microgels

Similar to the preparation of AgNPs/P(NIPAM-co-DMA) [37].

the hybrid microgels were prepared via a two-step process. First,
P(NIPAM-co-DMA) microgels were synthesized by a dispersion
polymerization in aqueous solution. Second, AuNPs/P(NIPAM-co- Fig. 1. A typical TEM image of AuNPs/P(NIPAM-co-DMA) hybrid microgels.
DMA) hybrid microgels were obtained by an in situ reduction of
AuCl4 ions incorporated into the microgels using NaBH4 as a 2+
PBA-AuNPs/P(NIPAM-co-DMA)+ Hg ions
reducing agent. A typical TEM image for the as-prepared AuNP- PBA-AuNPs/P(NIPAM-co-DMA)
entrapped microgels is shown in Fig. 1. Clearly, the spheric AuNPs 0.6 AuNPs/P(NIPAM-co-DMA)
Absorbance

PBA
with an average diameter of 4.0 0.5 nm were homogeneously
distributed within the P(NIPAM-co-DMA) microgels, and their 0.4
weight percentage content was calculated to be 19.8% based on
TGA weight loss, indicating the successful synthesis of AuNP-
embedded hybrid microgels. This should be due to the electrostatic 0.2
interaction between AuCl4 ions and protonated tertiary amine
groups as well as the coordination of Au atoms with the nitrogen
0.0
atoms in the polymer chains. The UVvis absorption spectrum of
AuNPs/P(NIPAM-co-DMA) hybrid microgels exhibits a distinctive 300 400 500 600 700 800
surface plasmon resonance (SPR) peak at 524 nm (Fig. 2), which is Wavelength / nm
assigned to the characteristic absorption band of AuNPs. In addi-
tion, the SPR band position as well as the absorption intensity had Fig. 2. UVvis absorption spectra of PBA, AuNPs/P(NIPAM-co-DMA), PBA-
AuNPs/P(NIPAM-co-DMA), and PBA-AuNPs/P(NIPAM-co-DMA) with Hg2+ ions.
little change, even after the hybrid microgels was stored for 50
days at room temperature (data not shown), indicating the hybrid
microgels was very stable. with AuNPs, the hybrid microgels may also possess similar prop-
erties. DLS was used to measure the Rh  of the hybrid microgels
3.2. Thermo- and pH-responsive properties of hybrid microgels at various temperatures and the results are displayed in Fig. 3. As
expected, the Rh  gradually decreased with increasing temper-
It is well known that P(NIPAM-co-DMA) microgels have ature at different pHs, indicating that AuNPs/P(NIPAM-co-DMA)
excellent thermo- and pH-responsive characters and display a hybrid microgels maintained the thermo-responsive property.
responsive swelling-shrinking behavior[37,38]. After embedded From Fig. 4, we can nd that as the pH was adjusted from 4.0 to
770 Y. Tang et al. / Sensors and Actuators B 228 (2016) 767773
150
pH 4.0
140 pH 7.4
pH 10.0
<Rh> / nm
130
120
110
20 25 30 35 40 45
o
T/ C
Fig. 3. Temperature dependence of the average hydrodynamic radius (Rh ) for
AuNPs/P(NIPAM-co-DMA) hybrid microgels with pH 4.0, 7.4 and 10.0. The concen-
tration of AuNPs/P(NIPAM-co-DMA) is 0.15 mg mL1 .
145
140
<Rh> / nm
135
130
125
4 6 8 10
pH
Fig. 4. pH dependence of the average hydrodynamic radius (Rh ) for
AuNPs/P(NIPAM-co-DMA) hybrid microgels at 20 C. The concentration of
AuNPs/P(NIPAM-co-DMA) is 0.15 mg mL1 .
10.0, the Rh  decreased from 142.5 to 130.1 nm, indicating the
pH-responsive behavior of the hybrid microgels. When the pH of
external environment is increased, the amino groups of PDMA seg-
ments are progressively deprotonated, which is responsible for the
collapse of the hybrid microgels and the decrease in Rh  [39].
3.3. The uorescence quenching of PBA by
AuNPs/P(NIPAM-co-DMA)
PBA molecule is an important uorescent probe because its
outstanding optical properties, such as high quantum yield and
long uorescence lifetime. It can interact with various molecules
via stacking, strong hydrogen bonding or electrostatic attrac-
tion, which make it extensively used as probes for the distinct
elds of study [4042]. Fig. 5 shows the uorescence emission
spectra of PBA with increasing concentrations of AuNPs/P(NIPAM-
co-DMA). Upon addition of the hybrid microgels to PBA solution,
the uorescence intensity of PBA signicantly decreased, whereas
its maximum emission peak did not show obvious shift. For com-
parison, we also recorded the uorescence spectra of PBA in the
presence of 0.12 mg mL1 P(NIPAM-co-DMA) microgels. As shown
in Fig. S1 in Supporting information, the uorescence intensity
shows a slight increase. These results suggest that the PBA uores-
cence was quenched more likely through the interactions between
AuNPs and PBA molecules [43]. In the present work, the electro-
static interaction and hydrogen bond between carboxyl units of
the PBA and amine groups of the DMA made PBA molecules dif-
fuse through the P(NIPAM-co-DMA) microgel network and reach
to AuNP surfaces. The efcient energy transfer from conned PBA
FL intensity / a.u.
12.0
2400
8.0
F0/F
1800 4.0
0
0.0
0.00 0.05 0.10 0.15 0.20
1200 [Hybrid microgels] / mgmL
-1
-1
0.2 mg mL
600
0
375 400 425 450 475 500
Wavelength / nm
Fig. 5. Fluorescence emission spectra of PBA with increasing concentrations of
AuNPs/P(NIPAM-co-DMA) with ex = 346 nm. From top to bottom: 0, 0.02, 0.04, 0.06,
0.08, 0.10, 0.12, 0.14, 0.16, 0.18, and 0.20 mg mL1 . Inset: dependence of F0 /F on the
hybrid microgel concentration, where F0 and F represent the uorescence intensity
of PBA at 398 nm in the absence and presence of the hybrid microgels.
to AuNPs resulted in the uorescence quenching. The more the con-
centration of hybrid microgels, the more PBA molecules interacted
with AuNPs, exhibiting a higher-efciency uorescence quenching.
A linear relationship between the F0 /F ratio and the hybrid microgel
concentration was found in the low concentration range (inset of
Fig. 5). However, when further increasing the hybrid microgel con-
centration, the relationship displayed an upward deviation from
the linearity, which was associated with the superquenching ef-
ciency of AuNPs [32]. As the hybrid microgel concentration was
0.20 mg mL1 , the quenching efciency of PBA was over 92% rela-
tive to the uorescence of unbound pure PBA. This superquenching
efciency is also required to maximize the uorescent change and
achieve an efcient turn-on detection of small amounts of analyte.
3.4. Detection of Hg2+ based on PBA-AuNPs/P(NIPAM-co-DMA)
Next, we measured the uorescence changes of PBA-
AuNPs/P(NIPAM-co-DMA) upon addition of varied amounts of Hg2+
ions. As shown in Fig. 6A, the uorescence intensity increased
successively in the Hg2+ concentration range from 0 to 3.04 M,
whereas the spectral widths and maximum emission wavelengths
did not show obvious change. When the concentration of Hg2+ was
3.04 M, the uorescence recovered to 83.7% of the initial intensity
of pure PBA, and the uorescence intensity increased 16-fold rela-
tive to the quenched PBA. A control experiment was performed in
the absence of AuNPs/P(NIPAM-co-DMA). There was no signicant
change in the emission spectrum of PBA. To understand the mech-
anism of the uorescence recovery, we investigated the changes of
the SPR band of PBA-AuNPs/P(NIPAM-co-DMA) dispersion in the
absence and presence of Hg2+ ions. As shown in Fig. 2, the SPR peak
shifted from 524 to 518 nm upon the addition of 1.6 M Hg2+ , and
the color of the solution had little change. Similar results were pre-
viously reported by Radhakumary et al. [44] and Morris et al. [45]
for the interaction of AuNPs with Hg2+ ions, where the SPR band
of AuNPs in the prescence of Hg2+ shrifted to a short wavelength
results from the formation of a core/shell structure. In addition,
Hg2+ ions most likely form stable complexes with the amine groups
in the PDMA segments. These collective results suggest that Hg2+
ions displace the PBA molecules attached on the AuNP surfaces,
leading to a signicant enhancement in the uorescence. On the
basis of the restored uorescence, a uorescent turn-on method
for the determination of Hg2+ was proposed.
To optimize the uorescence restoration of PBA for sensing Hg2+ ,
the effect of pH in a range from 6.0 to 10.0, incubation time and tem-
perature were investigated. The uorescence quenching efciency
of PBA in the presence of AuNPs/P(NIPAM-co-DMA) was higher in
 Y. Tang et al. / Sensors and Actuators B 228 (2016) 767773 771

6.0
2000 A PBA-AuNPs/P(NIPAM-co-DMA) + other ions

FL intensity / a.u.
c Hg
2+
2+
PBA-AuNPs/P(NIPAM-co-DMA) + other ions + Hg ions
5.0
1500
4.0

F/F0
1000
3.0
500
2.0
0
375 400 425 450 475 500
Wavelength / nm 1.0

20.0 0.0 2+ 2+ 2+ 2+ 2+ 2+ 3+ 3+ 2+ 2+ 2+ + + +
6.0 blank Hg Co Ba Pb Cu Fe Fe Al Mg Ca Zn Ag K Na
B
OF-F0P/F0

4.5

15.0 3.0 Fig. 7. Fluorescence intensity ratio (F/F0 ) of PBA-AuNPs/P(NIPAM-co-DMA) with or

1.5 without Hg2+ (1.28 M) with the coexistence of competing metal ions (12.8 M).
OF-F0P/F0

0.0

10.0 0.0 0.4 0.8 1.2 1.6

Hg2+/ M Table 2
Analytical results for the detection of Hg2+ in drinking water.
5.0
Sample Hg2+ added/nM Hg2+ found/nM Recovery (%) mean RSD, n = 3

Sample-1 160 165.8 4.50 103.6 2.8

0.0
0.0 0.8 1.6
Hg2+/ M
Fig. 6. (A) Fluorescence spectra of PBA-AuNPs/P(NIPAM-co-DMA) system in the
presence of increasing concentrations of Hg2+ ions with ex = 346 nm. The concen-
tration of Hg2+ ions in samples (solid curves) is 0, 0.16, 0.32, 0.48, 0.64, 0.80, 0.96,
1.12, 1.28, 1.44, 1.60, 1.76, 1.92, 2.08, 2.24, 2.40, 2.56, 2.72, 2.88, 3.04 M, respec-
tively. (B) A relationship between the uorescence intensity ratios [(F F0 )/F0 ] and
the concentrations of Hg2+ ions, where F and F0 represent the uorescence intensity
at 398 nm in the presence of different concentrations of Hg2+ and the absence of
Hg2+ , respectively.
the weak acid solutions than in alkaline media, maybe resulting
from the strong electrostatic interaction in weakly acid solutions.
However, the pH had little effect on the uorescence restoration of
PBA by Hg2+ ions. The higher quenching efciency is favorable for
the uorescence turn-on analysis of Hg2+ ions [43]. In addition, the
swollen hybrid microgels facilitated the analyte to be quickly trans-
ported within the polymer networks and close to or detach from the
AuNPs. For these reasons, the pH of 6.0 was adopted in the further
experiment. From Fig. S2 (Supporting information), we can nd that
the relative uorescence intensity decreased with temperature,
implying that it is not conducive to recover the uorescence of PBA
at high temperature. So, the subsequent experiments were carried
out at room temperature. At room temperature, time-dependent
measurements on the uorescence of PBA were conducted. Upon
the addition of AuNPs/P(NIPAM-co-DMA), the uorescent inten-
sity of PBA was dramatically decreased, and then reached a steady
value within 15 min (Fig. S3 in Supporting information). This u-
orescence was recovered within 3 min upon the addition of Hg2+
ions and then kept stable (Fig. S4 in Supporting information). Based
on the above two points, an incubation time of 30 min was selected
in the subsequent experiments.
Under the optimized experiment conditions, we investigated
the sensitivity of the proposed probe for Hg2+ in aqueous solu-
tion. The uorescence intensity ratios ((F F0 )/F0 ) was plotted
against the Hg2+ concentration as shown in Fig. 6B, in which F
and F0 represent the uorescence intensity at 398 nm in the pres-
ence of different concentrations of Hg2+ and the absence of Hg2+ ,
respectively. A linear relationship was observed between the val-
ues of (F F0 )/F0 and the concentration of Hg2+ ion over the range
of 0.161.6 M (inset in Fig. 6B). The calibration curve could be
expressed as (F F0 )/F0 = 0.252 + 3.827cHg2+ (c: M) with a cor-
Sample-2 320 321.6 12.7 100.5 4.0
2.4 3.2 Sample-3 480 486.1 20.8 101.3 4.3
Sample-4 640 612.3 16.1 95.70 2.5
Sample-5 800 800.8 22.3 100.1 2.8
*
The standard deviation of each sample was obtained by three measurements.
relation coefcient of R2 = 0.988. The detection limit was 31 nM,
estimated according to the 3/k, where  represents the stan-
dard deviation of eleven blank measurements and k is the slope
of the calibration curve. The present probe for Hg2+ is compared
with other advance optical methods (Table 1). The results show
that the linear range and the detection limit of our present probe
were comparable and even better than those of other probes. More
importantly, the PBA-AuNPs/P(NIPAM-co-DMA) exhibits high sta-
ble.
To assess the selectivity of the as-formed composites for Hg2+ ,
various environmentally relevant metal ions, including Co2+ , Ba2+ ,
Pb2+ , Cu2+ , Fe2+ , Fe3+ , Al3+ , Mg2+ , Ca2+ , Zn2+ , Ag+ , Na+ and K+ ions,
were examined under identical conditions (Fig. 7). Clearly, these
ions (12.8 M) did not cause any signicant changes in the uores-
cence intensity. On the other hand, the addition of Hg2+ (1.28 M)
resulted in a noticeable enhancement of F/F0 ratio, indicating the
present system exhibited high selectivity for the detection of Hg2+
ions. The effects of various coexisting ions on the uorescence
intensities of dispersion were further investigated by mixing the
interfering ions with Hg2+ ions. It can be seen that the dispersion
had similar behavior to that containing pure Hg2+ , suggesting that
the coexistence of selected metal ions did not interfere with the
selectivity of the developed Hg2+ probe.
To test the practicality of the proposed method, this turn-on
optical Hg2+ ions probe was used to determine the concentration
of Hg2+ in drinking water by the standard addition method. From
Table 2, it can be seen that the detected concentrations of Hg2+
were close to those of the added Hg2+ ions. In addition, the recov-
eries were in the range of 95.7103.6% and the relative standard
deviations were all less than 5.0%. The results indicated that the
developed probe has satisfactory accuracy and reproducibility.
4. Conclusions
In summary, we designed a sensitive and turn-on uo-
rescent method for the determination Hg2+ ions based on
PBA-AuNPs/P(NIPAM-co-DMA) composites. The AuNPs/P(NIPAM-
772 Y. Tang et al. / Sensors and Actuators B 228 (2016) 767773
co-DMA) hybrid microgels exhibited thermo- and pH-responsive
properties and great stability. The uorescence of PBA was
quenched signicantly by AuNPs/P(NIPAM-co-DMA) through
energy transfer from PBA to AuNPs. Hg2+ ions displace the PBA
molecules attached on the AuNP surfaces that led to the uo-
rescence recovery of PBA. The uorescence turn-on response is
linearly proportional to the concentration of Hg2+ in the range of
0.161.6 M with detection limit 31 nM. Moreover, the probe can
recognize Hg2+ in presence of other abundant metal ions with high
selectivity. The present results offer valuable information and cre-
ate a good platform for detecting of Hg2+ . Thus, this strategy could
be used as a general approach to construct other microgels based
uorescence probes.
Acknowledgements
The nancial support from the Natural Science Foundation
of Anhui Province, China (11040606M61), the Natural Science
Foundation of the Anhui Higher Education Institutions of China
(KJ2011A137) and the Innovation Funds of Anhui Normal Univer-
sity is gratefully acknowledged.
Appendix A. Supplementary data
Supplementary data associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.snb.2016.01.112.
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Biographies
Yecang Tang is an associated professor of College of Chemistry and Materials Sci-
ence, Anhui Normal University, China. He obtained his Ph.D. from University of
Science and Technology of China in 2008 and M.S. from Anhui Normal University in
2002. Her research interests include the development of nanocomposite materials
for sensing applications and polymer science.
Yi Ding obtained his B.Sc. degree from Anhui University of Technology in 2011. He is
currently studying for his M.Sc. degree in College of Chemistry and Materials Science,
Anhui Normal University, China. He is engaged in the synthesis of nanocomposites
for sensing applications.
Ting Wu is a teacher of Urban Construction College of Anhui Jianzhu University,
China. She obtained her B.Sc. and M.Sc. degree from Anhui Normal University in 2011
and 2014, respectively. Her research interests focus on the design and fabrication of
nanomaterials.
Liying Lv received her B.Sc. degree from Guangdong University of Petrochemi-
cal Technology in 2013. She is currently pursuing his M.Sc. degree in College of
Chemistry and Materials Science, Anhui Normal University. Her scientic interest is
developing uorescent probes based on nanocomposites.
Zhicheng Yan is an undergraduate student in the College of Chemistry and Materials
Science, Anhui Normal University.