Microchimica Acta (2020) 187:185

https://doi.org/10.1007/s00604-020-4158-2

ORIGINAL PAPER

Highly selective fluorimetric and colorimetric sensing of mercury(II)

by exploiting the self-assembly-induced emission
of 4-chlorothiophenol capped copper nanoclusters
Hai-Bo Wang 1 & Hong-Yu Bai 1 & Ya-Shu Wang 1 & Tian Gan 1 & Yan-Ming Liu 1

Received: 15 August 2019 / Accepted: 13 February 2020

# Springer-Verlag GmbH Austria, part of Springer Nature 2020

Abstract
A highly stable copper nanoclusters (CuNC) carrying 4-chlorothiophenol as a protective ligand is described. They display self-
assembly-induced emission with excitation/emission maxima at 330/605 nm even in neutral or alkaline aqueous environment.
The fluorescence of these CuNC is quenched by Hg(II). Quenching is mainly ascribed to the formation of a complex formed via
Hg-S bonding between the Hg(II) ions and the ligand. This destroys the ordered architectures of the assembled CuNC. The assay
enables Hg(II) to be determined with good sensitivity and a linear response ranging from 1 to 500 nM Hg(II) with a 0.3 nM limit
of detection. In addition, the method was implemented in a test strip (which undergoes a color change from red to blue) that can be
used for visual determination of Hg(II) in complex environmental water samples.

Keywords Aggregation-induced emission enhancement . Luminescent test strips . Metal nanoclusters . Hg(II) ions
determination . Fluorimetric nanoprobe . Colorimetric assay . Visual detection . Coordination complex . Fluorescence quenching

Introduction is relatively cheap, abundant, and readily available [6–9]. Many

As compared with organic fluorophores and semiconductor
quantum dots, fluorescent metal nanoclusters (including Au
nanoclusters, Ag nanoclusters, and Cu nanoclusters), as a novel
luminescent nanomaterial, have attracted much research inter-
ests in many fields in the past few years [1–5]. The fluorescent
metal nanoclusters possess many superior properties, including
excellent luminescence, controllable emission wavelength, large
Stokes shift, good biocompatibility, and high photo-stability. In
comparison with gold nanoclusters and silver nanoclusters, the
presursor (Cu2+ ions) used for the copper nanoclusters (CuNC)
Electronic supplementary material The online version of this article
(https://doi.org/10.1007/s00604-020-4158-2) contains supplementary
material, which is available to authorized users.
* Hai-Bo Wang
wanghaibohn@163.com
* Tian Gan
gantianxynu@163.com
1
College of Chemistry and Chemical Engineering, Institute for
Conservation and Utilization of Agro-bioresources in Dabie
Mountains, Xinyang Normal University, Xinyang 464000, People’s
Republic of China
fluorescent CuNC have been reported based on different types
of protecting ligands or stabilizers (such as DNA, peptides, pro-
teins, thiols, and so on) [6–11]. These fluorescent CuNC are also
utilized to construct fluorescent, electrochemical,
electrochemiluminescent, and photoelectrochemical sensors or
biosensors for the sensitive detection of DNA, protein, metal
ions, small molecules, and enzymes activity [11–16].
Nevertheless, the stability of these fluorescent CuNC is poor
because CuNC is inherently prone to aggregate and the surface
is easily oxidized on exposure to air. Thus, to develop bright and
stable fluorescent, CuNC is still highly desirable.
Aggregation-induced emission (AIE) enhancement of
thiolated CuNC have become a promising class of luminescent
nanomaterials, due to their strong luminescence emission, high
fluorescence quantum yield, long fluorescence lifetimes, and
inherent biocompatibility [17, 18]. The AIE property is ascribe
to that high compactness of assembled thiolated CuNC
prevented the intramolecular vibration and rotation of the
protecting/capping ligands, leading to the enhancement of emis-
sion intensity [11, 17, 18]. Up to now, many different thiolated
capping ligands have been utilized to prepare AIE-based CuNC
[17–21]. For example, Jia et al. have reported CuNC with AIE
feature by employing D-penicillamine as the capping ligands
[18]. The CuNC is explored for pH stimuli-responsive, H2O2
 185 Page 2 of 9
and glucose biosensing. Qian and co-workers have reported that
glutathione (GSH) can act as the reductant and the capping agent
for the preparation of AIE-based CuNC [19]. And the AIE state
of CuNC is reversible and controlled through the adding water
or aprotic solvents. Zhao’s group have exploited a GSH-CuNC-
based AIE enhancement for the detecting and imaging of Zn2+
in MGC-803 cells [20]. Nevertheless, retaining the strong lumi-
nescence of CuNC in neutral or alkaline solution is still a chal-
lenge [22]. These CuNC only exhibit strong fluorescence signal
in acidic solution and the fluorescence signal is lost in neutral or
alkaline solution. Furthermore, the luminescence is extremely
susceptible to the temperature changes and the existence of ox-
idizing agent (e.g., H2O2).
Yang’s group has reported strong and tunable red emission
of self-assembly-induced emission (S-AIE) CuNC by utiliz-
ing aromatic thiols instead of alkylthiol ligands as the capping
ligands [23]. Due to its excellent emission intensity and photo-
stability, the S-AIE CuNC architectures have been exploited
for the fabrication of CuNC-based white light-emitting diodes
(WLEDs) devices [23]. The bright emission intensity is main-
ly attributed to the well-ordered nano-structures of the self-
assembly CuNC which decreased the non-radiative relaxation
by blocking the vibration and rotation of capping ligands [21].
Zhao and co-workers have reported stable and luminescent
self-assembly CuNC-based AIE particles using 4-
methylthiophenol as the capping ligands [22]. The CuNC is
also applied for the evaluation of the β-galactosidase activity
in a neutral solution by controllable hydrophobicity interac-
tion. Han et al. have reported highly bright S-AIE CuNC by
using 2, 3, 5, 6-tetrafluorothiophenol as both the capping li-
gands and the reducing agents [24]. On the basis of the S-AIE
CuNC, fluorescent assay and test strips are constructed for the
determination of histamine content in food samples. However,
the exploration of S-AIE CuNC-based sensing or biosensing
methods is still at its very early stage.
In this work, highly luminescent and stable S-AIE CuNC
was synthesized by using 4-chlorothiophenol as the capping
ligands and hydrazine hydrate as the reducing agents.
Interestingly, it was found that the fluorescence signal was
obviously quenched by Hg2+ ions. On the basis of the phe-
nomenon, a fluorescent strategy was designed for the determi-
nation of Hg2+ ions. Furthermore, the S-AIE CuNC was
employed to fabricate novel fluorescent test strips for the col-
orimetric determination of Hg2+ ions content in environmental
water samples visually.
Experimental
Materials and reagents
Cu(NO3)2•3H2O, 4-chlorothiophenol, 50% hydrazine hydrate
(NH2NH2), ethanol absolute, and other chemical reagents
Microchim Acta (2020) 187:185
were analytical reagent grade and purchased from Shanghai
Aladdin Bio-Chem Technology Co., Ltd. (Shanghai, China)
(http://www.aladdin-e.com). All water (used in this work) was
ultrapure water (with electric resistance > 18.2 MΩ cm) by
using a Millipore Milli-Q water purification system (MA,
USA, http://www.merckmillipore.com).
Apparatus
All fluorescent measurements were recorded on a F-7000
spectrophotometer (Hitachi Co. Ltd., Japan). The fluorescence
excitation wavelength was set at 330 nm; the emission spectra
were performed from 500 nm to 650 nm. UV-vis absorption
spectra were carried out on a U-3900H spectrophotometer
(Hitachi Co. Ltd., Japan). Transmission electron microscopy
(TEM) images were performed by using a Tecnai G2 F20 S-
TWIN field emission transmission electron microscopy. X-ray
photoelectron spectroscopy (XPS) was recorded on a K-Alpha
spectrometer. The hydrodynamic sizes of CuNC were ana-
lyzed by dynamic light scattering (DLS) on a Mastersizer
3000E particle size analyzer (UK).
Preparation of S-AIE CuNC
In brief, 10 mg (0.04 mmol) Cu(NO3)2•3H2O, 1 mL 50%
NH2NH2 were mixed with 20 mL ultrapure water with vigor-
ous stirring. After stirring for 10 min, 58 mg (0.4 mmol) of 4-
chlorothiophenol was added to the above mixture. After stir-
ring for another 30 min at room temperature, the solution
changed to yellow turbid. Then, the solution was under
centrifuging treatment (20,000 rpm) for 10 min. The precipi-
tate was collected and washed 3 times with ethanol to remove
the excessive other reactants. Then, the product was dispersed
in ethanol by ultrasonic treatment for 30 min. Finally, the S-
AIE CuNC (~ 0.5 mg mL−1) was stored at 4 °C in the refrig-
erator for further use.
Procedures for fluorescent determination of Hg2+ ions
For Hg2+ determination, 50 μL 0.2 mg mL−1 S-AIE CuNC,
10 μL Hg2+ ions solution (different concentrations), and
40 μL 25 mM phosphate buffered solution (pH 7.0) were
mixed and incubated at room temperature for 5 min. Then,
the fluorescent intensity at 605 nm was collected immediately.
The selectivity toward Hg2+ was evaluated by testing other
cations including Ni2+, Mg2+, Cd2+, Cu2+, Mn2+, Zn2+, Ba2+,
and Pb2+ ions.
Procedures for determination of Hg2+ in real samples
Tap water, lake water, and wastewater samples were collected
from our laboratory, Nanwan Lake, industry sewerage in Shi
River, respectively. All the water samples were centrifuged
Microchim Acta (2020) 187:185
(12,000 rpm) and filtered through a 0.45 μm cellulose mem-
brane to remove oils and suspended particles before test.
Different concentration levels of Hg2+ ions in 10 mM PBS
(pH 7.0) were spiked into the tap water samples, lake water
samples, and waste water samples. The resulting solutions
were further mixed with S-AIE CuNC solutions. After 5 min
incubation, the fluorescent intensity at 605 nm was collected
with the excitation wavelength of 330 nm.
Preparation of the test strips and colorimetric
determination of Hg2+ ions
Briefly, a kind of cotton fiber absorbent pad was used for the
preparation of the test strips. The absorbent pad was firstly cut
into square pieces with scissors (e.g. 1 × 1 cm). Then, each of
the square pieces was immersed into 500 μL of
0.25 mg mL−1 S-AIE CuNC solution. After being dried, the
S-AIE CuNC-based fluorescence test strips were obtained.
Then, various concentrations of Hg2+ ions solution were
added onto the test strips (5 times, 100 μL per time). And
the fluorescent test strips were illuminated by 365 nm of UV
light. At last, the photographs of the color changes of the test
strips were recorded by a camera.
Results and discussion
Design principle of the fluorimetric and colorimetric
assay
The design principle of the fluorimetric and colorimetric assay
is shown in Scheme 1. Highly luminescent and stable S-AIE
CuNC was synthesized by using 4-chlorothiophenol (SR) as
the capping/protecting ligands and NH2NH2 as the reducing
agents. The CuNC can be regarded as a particular Cu-SR com-
plex substance, which was formed on the surface of Cu(0) core.
The size of CuNC is approximately spherical and the diameter
is about 3–5 nm. Afterwards, the aggregation of the shell Cu-
SR complex substance on the different clusters can form or-
dered nanorod-like structure, and improve the fluorescence in-
tensity through the self-assembly-induced emission mecha-
nism. The self-assembly of CuNC is mainly driven by van
der Waals attraction and dipolar attraction [23]. The assembled
induced emission property is mainly attributed to that the well-
ordered nano-structures of the self-assembly CuNC can de-
crease the non-radiative relaxation by blocking the vibration
and rotation of protecting ligands [23]. According to Scheme
1, the S-AIE CuNC possesses high fluorescence signal.
Interestingly, it is found that the fluorescent signal is obviously
quenched in the presence of Hg2+ ions. Presumably, it is mainly
attributed to the formation of the stable complexes by Hg-S
bonds between the protecting ligand (4-chlorothiophenol) and
Hg2+ ions. The solubility product constants (Ksp) of HgS (3 ×
Page 3 of 9 185
10−52) are much smaller than that of CuS (8 × 10−36) [25], sug-
gesting that Hg-SR complexes are much more stable than that
of Cu-SR complexes capped on the surface of Cu(0) core.
Thus, the ordered nanorod-like architectures of S-AIE CuNC
are destroyed due to the formation of the Hg-SR complex. The
protecting ligand (4-chlorothiophenol) is released far away
from the surface of Cu(0) core, leading to the Cu(0) core which
aggregated and formed larger ones. As a result, the fluorescence
intensity of CuNC is significantly decreased without the pro-
tection of scaffolds. On the other hand, the S-AIE CuNC is
utilized to fabricate new luminescent test strips for colorimetric
determination of Hg2+ levels visually. More interesting, the
color of test strips changes obviously from red to blue-purple
in the presence of Hg2+ ions. Thus, according to the changes of
the fluorescence intensity and the test strips color, rapid fluo-
rescent strategy and test strips have been designed for the sen-
sitive determination of Hg2+ ions.
Characterization of optical properties of S-AIE CuNC
Figure 1a displays the typical fluorescence excitation spec-
trum and emission spectrum of the S-AIE CuNC. While the
excitation wavelength was set to 330 nm, a strong red lumi-
nescence signal was obtained at 605 nm. Figure 1b and c show
the X-ray photoelectron spectroscopy (XPS) of the S-AIE
CuNC. From Fig. 1b, it is found that the CuNC showed the
Cu, C, Cl, and S elements, which are from the raw materials.
Additionally, as indicated in the high-resolution XPS spec-
trum of Cu 2p (Fig. 1c), the peaks at 952.9 and 933.1 eV
can be observed, and no characteristic satellite peaks appear
around 942 eV, suggesting that copper states in the S-AIE
CuNC mainly existed in the lower valence states Cu(0) and
Cu(I), rather than Cu(II). The FTIR spectra (Fig. 1d) of S-AIE
CuNC do not show the characteristic peaks of -SH around
2575 nm, proving the binding of protecting ligands (4-
chlorothiophenol) with the Cu core through the Cu-S bond.
Importantly, the fluorescence intensity (605 nm) of S-AIE
CuNC is almost unchanged as the solution pH ranges from
5.0 to 9.0 (Fig. S1A). Even though under the higher ion
strength (10 mM NaCl), the S-AIE CuNC has strong lumines-
cence signal (Fig. S1B). Additionally, it is found that the lu-
minescence intensity of the S-AIE CuNC is not obviously
decreased after stored at 4 °C for 10 months. These results
suggest that the S-AIE CuNC exhibits excellent stability and
has potential to be exploited as fluorescent probes for the
development of biosensing strategy.
Evaluation of the feasibility of the fluorescence assay
To evaluate the S-AIE CuNC-based fluorescence strategy for
Hg2+ determination, the fluorescence emission spectra of S-
AIE CuNC was studied under different experimental condi-
tions. As displayed in curve a of Fig. S2A, it is found that S-
 185 Page 4 of 9 Microchim Acta (2020) 187:185

Scheme 1 Schematic illustration of the fluorescent strategy and test strips for the selective determination of Hg2+ ions based on the highly stable S-AIE
CuNC

AIE CuNC had high luminescence signal at 605 nm in the
absence of Hg2+. When 10 μM Hg2+ is introduced into the
system (curve b of Fig. S2A), the fluorescence intensity of
CuNC decreased strongly. It may be ascribed to that the stable
Hg-SR complex is formed and disrupted the ordered architec-
tures of S-AIE CuNC, resulting in the fluorescence
quenching. Furthermore, the incubation time of Hg2+ with
CuNC is investigated. From Fig. S2B, it is obviously seen that
significant fluorescence quenching is observed within 1 min
in the presence of 10 μM Hg2+ and attained the equilibrium as
soon as within 3 min. As a result, the fluorescence intensity of
S-AIE CuNC is quenched by Hg2+ rapidly.

Fig. 1 A Fluorescence absorption

(a) and emission (b) spectrum of
the S-AIE CuNC. B XPS spectra
of S-AIE CuNC. C High-
resolution Cu (2p) XPS spectra of
S-AIE CuNC. D The FTIR spec-
tra of 4-chlorothiophenol (a) and
S-AIE CuNC (b)
Microchim Acta (2020) 187:185
Fig. 2 UV-vis absorption spectra of 500 μM 4-chlorothiophenol (a), S-
AIE CuNC (b), S-AIE CuNC +100 μM Hg2+ (c), and 500 μM 4-
chlorothiophenol +150 μM Hg2+ (d)
Mechanism for the fluorescence quenching of CuNC
by Hg2+
To better understand the fluorescence quenching mechanism of
Hg2+ induced the fluorescence quenching of S-AIE CuNC,
UV-vis absorbance spectra, TEM images, and dynamic light-
scattering measurements were further investigated. As shown in
Fig. 2, the S-AIE CuNC has significant absorption ranging
from 200 nm to 500 nm and a maximum absorption peak at
330 nm (curve b), which is remarkably different from the
protecting ligand of 4-chlorophenthiophene (curve a) (maxi-
mum absorption peak at 245 nm). While in the presence of
Hg2+, the characteristic peak of CuNC at 330 nm is disappeared
Fig. 3 a TEM images of S-AIE CuNC. b TEM images of S-AIE CuNC with Hg2+ ions
Page 5 of 9 185
and a new absorption peak is appeared at 245 nm (curve c in
Fig. 2), which is coincident with that of the 4-
chlorophenthiophene with Hg2+ (curve d in Fig. 2). It means
that the protecting ligand (4-chlorothiophenol) has greater co-
ordination affinity with Hg2+ to form stable Hg-SR complexes.
The TEM image (Fig. S3) of CuNC dispersion in ethanol clear-
ly illustrates that the sizes are at 3–5 nm in diameter. In addition,
the morphology of the S-AIE CuNC with and without Hg2+
was characterized by TEM images. From Fig. 3a, it can be seen
that the S-AIE CuNC shows nanorod-like structure with the
average width of about 30 nm and the average length of 200–
400 nm. As shown in Fig. 3b, TEM image indicates that or-
dered nanorod-like structure is destroyed and the size became
much larger (the average width of 60–140 nm, the average
length of 1000–1600 nm), when the S-AIE CuNC was in the
presence of Hg2+. Furthermore, dynamic light-scattering mea-
surement was utilized to determinate the hydrodynamic size
distribution of these S-AIE CuNC with and without Hg2+. As
shown in Fig. S4, the size of S-AIE CuNC becomes larger
when Hg2+ is present. The results are in agreement with the
result of the above TEM images. Taking these results together,
the Hg2+-induced the fluorescence quenching of S-AIE CuNC
is attributed to the formation of the Hg-SR complex, which
leads to the destruction of the ordered architectures of S-AIE
CuNC and the protecting ligand released and far away from the
surface of Cu(0) core, accompanying the Cu(0) core aggregated
and enlarged.
Analytical performance of the fluorescent strategy
for Hg2+ assay
In order to confirm the applicability of the strategy for Hg2+
assay, the fluorescence intensity changes of S-AIE CuNC
 185 Page 6 of 9 Microchim Acta (2020) 187:185

Fig. 4 a Fluorescence emission

spectra of the strategy in the
presence of increasing
concentrations of Hg2+ ions. b
The relationship between the
fluorescence intensity at
excitation/emission wavelengths
of 330/605 nm and the Hg2+ con-
centrations. c The calibration plot
between the fluorescence re-
sponse (F0/F) and Hg2+ concen-
trations. Where F0 and F were the
fluorescence intensity of S-AIE
CuNC without and with Hg2+
ions, respectively. d The selectiv-
ity of the system for Hg2+ assay.
The error bars are the standard
deviation of three repetitive
measurements

were tested by addition of a series of various concentrations of
Hg2+ ions. Figure 4a displays the typical fluorescence emis-
sion spectra of S-AIE CuNC with various concentrations of
Hg2+ ions. It is observed that the luminescence of S-AIE
CuNC quenched gradually along with the increase of Hg2+
ions concentration from 0 to 100 μM (curves from the top to
the bottom). As shown in Fig. 4b, the fluorescence intensity at
605 nm is remarkably quenched, when the Hg2+ concentra-
tions increase. The result suggested that the more Hg2+ pre-
sented, the more Hg-SR duplex formed, leading to more S-
AIE CuNC destruction and fluorescence quenching. From
Fig. 4c, the fluorescence response (F0/F) vs Hg2+ concentra-
tions indicate typical Stern-Volmer quenching behavior. It is
found that a linear relationship between fluorescence response
(F0/F) and Hg2+ concentrations (from 1 to 500 nM). On ac-
count of signal-to-noise (S/N) ratio of 3, the limit of detection
is calculated to be 0.3 nM, which is lower than some reported
methods [26–29]. The limit of detection is much lower than
the permitted maximum level (10 nM) for Hg2+ defined by the
United States (U.S.) Environmental Protection Agency (EPA)
in drinking water [28]. Furthermore, the method is superior to
other reported methods with better or comparable detection
limit and linear range [26–29], listed in Table 1.
To further study the selectivity of the fluorescent strategy,
we have investigated the fluorescent response of
10 μM Hg2+ and 100 μM other divalent metal ions (includ-
ing Ni2+, Mg2+, Cd2+, Cu2+, Mn2+, Zn2+, Ba2+, and Pb2+). As
indicated in Fig. 4d, it is observed that there is no obvious
fluorescence response in the presence of other divalent metal
ions, suggesting high selectivity for Hg2+ determination.
This is attributed to the stronger affinity between the Hg2+
ion and the protecting ligand (SR). The method was further

Table 1 Comparison of various

analytical methods for Hg2+ Materials used Method Linear range (μM) Limit of References
determination applied detection (nM)

Carbon dot Fluorescence 0–40 9.0 [26]

Folic-acid-capped gold nanoclusters Fluorescence 0.1–1 28 [27]
Gold nanoparticles Colorimetric 0.05–0.25 50 [28]
TPE-functionalized Fluorescence Not given 1000 [29]
benzothiazolium salts
Self-assembly copper nanoclusters Fluorescence 0.001–0.5 0.3 this work
Microchim Acta (2020) 187:185 Page 7 of 9 185

Table 2 Determination of Hg2+

content in tap water and lake Samples Spiked (nM) Found (nM) Recovery (%) RSD (n = 3, %)
water samples
Tap water 1 2.00 1.97 98.5 3.20
Tap water 2 20.00 20.56 102.8 2.86
Tap water 3 100.00 98.32 98.3 3.45
Lake water 1 2.00 2.06 103.0 2.74
Lake water 2 20.00 20.14 100.7 3.36
Lake water 3 100.00 99.18 99.2 2.97

applied to determine the Hg2+ ion in environmental water
samples. Table 2 displays the results of determination of
Hg2+ in tap and lake water samples, the recoveries are from
98.3 to 103.0%. Table S1 shows the results of determination
of Hg2+ content in waste water sample, the RSD is bet-
ter 4.2%. These results suggested the applicability of the
assay for Hg2+ determination in real samples.
Fabrication of test strips for visual determination
of Hg2+ ions
The test strips were constructed by soaking the cotton fiber
absorbent pad in S-AIE CuNC suspensions. Compared with
blank cotton pad (blue), the test strips based on the S-AIE
CuNC are red under the radiation of 365 nm UV light
(Fig. 5a). Importantly, after the test strips (in hermetic bag)
were stored at 4 °C for 5 months, it still maintained strong
luminescence and the color has no obvious changes under
365 nm UV light (Fig. 5b). However, it is observed that the
color of the test strips obviously vary from red to blue-purple
with the increase of Hg2+ ions concentrations (Fig. 5c). And
the lowest Hg2+ concentration observed is 0.1 μM, suggesting
that the lowest detectable concentration (for Hg2+ ions) of the
test strips can be as low as 0.1 μM visually, which is lower
than the permitted limit value (250 nM) of Hg2+ by the U.S.
EPA in polluted water industry [28]. As shown in Fig. 5d, the
color of test strips has no obvious changes compared with
other metal ions. Only in the presence of Hg2+ ions, the test
strips vary from red to blue. The color changes of the test strips
prove the excellent selectivity toward Hg2+ ions compared
with other metal ions. Therefore, the S-AIE CuNC-based test
strips can offer a convenient and cost-effective method for
sensitive determination of Hg2+ ions visually. The S-AIE
CuNC-based test strips were also applied for visual

Fig. 5 A Photographs of blank cotton fiber absorbent pad (a, c) and test AIE CuNC-based test strips for Hg2+ ions determination. D Photographs
strips based on S-AIE CuNC (b, d) under daylight (a, b) and 365 nm UV of test strips in the presence of 150 μM of Hg2+ and other metal ions
light (c, d). B Photographs of fluorescence test strips after stored in her-
metic bag for 5 months under 365 nm UV light. C Photographs of the S-
 185 Page 8 of 9
monitoring of the Hg2+ ions levels in tap water and lake water
samples. From Fig. S5, the test strips display an obvious color
gradient along with the Hg2+ ions levels increase, demonstrat-
ing the potential applications of test strips for real samples.
The UV light used for fluorescence excitation will be screened
off by UV absorbers and this will weaken the signal.
Conclusions
A novel fluorescence strategy and test strips have been dem-
onstrated for the determination of Hg2+ ion by using the ex-
cellent luminescence properties of the S-AIE CuNC. The as-
say mainly relies on the strong binding interaction of Hg2+
ions and the protecting ligand on the surface of the S-AIE
CuNC. The strategy has some distinct advantages. Firstly,
the S-AIE CuNC possesses high luminescence and photo-sta-
bility. Secondly, the method shows high selectivity for Hg2+
ions determination compared to other metal ions. Thirdly, the
preparation of the S-AIE CuNC-based test strips is convenient
and cost-effective. Finally, it can be utilized for the visual
assay of Hg2+ in real samples. Therefore, the novel strategy
may find applications for rapid determination of Hg2+ ions
level in aqueous biological and environmental samples.
However, it is a little time consuming to purify the CuNC.
Funding information This work was financially supported by the
National Natural Science Foundation of China (No. U1704153,
21864004), 1000 Talent Program of Zhongyuan, Plan for Young
Excellent Teachers in Universities of Henan Province (No.
2017GGJS100), Nanhu Scholars Program for Young Scholars of XYNU.
Compliance with ethical standards
Conflict of interest The authors declare that they have no com-
peting interests.
References
1. Zhang N, Si YM, Sun ZZ, Chen LJ, Li R, Qiao YC, Wang H (2014)
Rapid, selective, and ultrasensitive fluorimetric analysis of mercury
and copper levels in blood using bimetallic gold-silver nanoclusters
with “silver effect”-enhanced red fluorescence. Anal Chem 86:
11714–11721
2. Zhang LL, Zhao JJ, Zhang H, Jiang JH, Yu RQ (2013) Double
strand DNA-templated copper nanoparticle as a novel fluorescence
indicator for label-free detection of polynucleotide kinase activity.
Biosens Bioelectron 44:6–9
3. Si YM, Sun ZZ, Zhang N, Qi W, Li SY, Chen LJ, Wang H (2014)
Ultrasensitive electroanalysis of low-level free microRNAs in
blood by maximum signal amplification of catalytic silver deposi-
tion using alkaline phosphatase-incorporated gold nanoclusters.
Anal Chem 86:10406–10414
4. Tian X, Kong XJ, Zhu ZM, Chen TT, Chu X (2015) A new label-
free and turn-on strategy for endonuclease detection using a DNA-
silver nanocluster probe. Talanta 131:116–120
Microchim Acta (2020) 187:185
5. Guo LY, Tang T, Hu LS, Yang MH, Chen X (2017) Fluorescence
assay of Fe (III) in human serum samples based on pH dependent
silver nanoclusters. Sensors Actuators B Chem 241:773–778
6. Wang HB, Zhang HD, Chen Y, Liu YM (2015) A fluorescent bio-
sensor for protein detection based on poly(thymine)-templated cop-
per nanoparticles and terminal protection of small molecule-linked
DNA. Biosens Bioelectron 74:581–586
7. Huang Y, Feng H, Liu WD, Zhou YY, Tang C, Ao H, Zhao MZ,
Chen GL, Chen JR, Qian ZS (2016) Luminescent aggregated cop-
per nanoclusters nanoswitch controlled by hydrophobic interaction
for real-time monitoring of acid phosphatase activity. Anal Chem
88:11575–11583
8. Wang HB, Zhang HD, Chen Y, Huang KJ, Liu YM (2015) A label-
free and ultrasensitive fluorescent sensor for dopamine detection
based on double-stranded DNA templated copper nanoparticles.
Sensors Actuators B Chem 220:146–153
9. Qing T, Zhang K, Qing Z, Wang X, Long C, Zhang P, Hu H, Feng B
(2019) Recent progress in copper nanocluster-based fluorescent
probing: a review. Microchim Acta 186:670
10. Wang YL, Cui YY, Liu R, Wei YT, Jiang XL, Zhu HR, Gao L, Zhao
YL, Chai ZF, Gao XY (2013) Blue two-photon fluorescence metal
cluster probe precisely marking cell nuclei of two cell lines. Chem
Commun 49:10724–10726
11. Wang HB, Chen Y, Li N, Liu YM (2017) A fluorescent glucose
bioassay based on the hydrogen peroxide-induced decomposition
of a quencher system composed of MnO2 nanosheets and copper
nanoclusters. Microchim Acta 184:515–523
12. Liu R, Wang CQ, Hu JY, Su YY, Lv Y (2018) DNA-templated
copper nanoparticles: versatile platform for label-free bioassays.
Trends Anal Chem 105:436–452
13. Wang Y, Zhang X, Zhao L, Bao T, Wen W, Zhang XH, Wang SF
(2017) Integrated amplified aptasensor with in-situ precise prepara-
tion of copper nanoclusters for ultrasensitive electrochemical detec-
tion of microRNA 21. Biosens Bioelectron 98:386–391
14. Zhao M, Chen AY, Huang D, Zhuo Y, Chai YQ, Yuan R (2016) Cu
nanoclusters: novel electrochemiluminescence emitters for
bioanalysis. Anal Chem 88:11527–11532
15. Lv S, Li Y, Zhang K, Lin Z, Tang DP (2017) Carbon dots/g-C3N4
nanoheterostructures-based signal-generation tags for
photoelectrochemical immunoassay of cancer biomarkers coupling
with copper nanoclusters. ACS Appl Mater Interfaces 9:38336–
38343
16. Mei LP, Jiang XY, Yu XD, Zhao WW, Xu JJ, Chen HY (2018) Cu
nanoclusters-encapsulated liposomes: toward sensitive liposomal
photoelectrochemical immunoassay. Anal Chem 290:2749–2755
17. Jia X, Li J, Wang EK (2013) Cu nanoclusters with aggregation
induced emission enhancement. Small 9:3873–3879
18. Jia X, Yang X, Li J, Li DY, Wang EK (2014) Stable Cu
nanoclusters: from an aggregation-induced emission mechanism
to biosensing and catalytic applications. Chem Commun 50:237–
239
19. Huang YY, Liu WD, Feng H, Ye YT, Tang C, Ao H, Zhao MZ,
Chen GL, Chen JR, Qian ZS (2016) Luminescent nanoswitch based
on organic-phase copper nanoclusters for sensitive detection of
trace amount of water in organic solvents. Anal Chem 88:7429–
7434
20. Lin LY, Hu YF, Zhang LL, Huang Y, Zhao SL (2017)
Photoluminescence light-up detection of zinc ion and imaging in
living cells based on the aggregation induced emission enhance-
ment of glutathione-capped copper nanoclusters. Biosens
Bioelectron 94:523–529
21. Wu Z, Liu J, Gao Y, Liu H, Li T, Zou H, Wang Z, Zhang K, Wang Y,
Zhang H, Yang B (2015) Assembly-induced enhancement of cu
nanoclusters luminescence with mechanochromic property. J Am
Chem Soc 137:12906–12913
Microchim Acta (2020) 187:185
22. Zhao MZ, Qian ZS, Zhong MT, Chen ZT, Ao H, Feng H (2017)
Fabrication of stable and luminescent copper nanocluster-based
AIE particles and their application in β-galactosidase activity assay.
ACS Appl Mater Interfaces 9:32887–32895
23. Ai L, Jiang WR, Liu ZY, Liu JL, Gao Y, Zou HY, Wu ZN, Wang
ZG, Liu Y, Zhang H, Yang B (2017) Engineering a red emission of
copper nanocluster self-assembly architectures by employing aro-
matic thiols as capping ligands. Nanoscale 9:12618–12627
24. Han AL, Xiong L, Hao SJ, Yang YY, Li X, Fang GZ, Liu JF, Pei Y,
Wang S (2018) Highly bright self-assembly copper nanoclusters: a
novel photoluminescent probe for sensitive detection of histamine.
Anal Chem 90:9060–9067
25. Wang HB, Chen Y, Li Y, Zhang HD, Cao JT (2015) A rapid,
sensitive and label-free sensor for hg(II) ions detection based on
blocking of cysteine-quenching of fluorescent poly(thymine)-tem-
plated copper nanoparticles. RSC Adv 5:94099–94104
26. Zhao JJ, Huang MJ, Zhang LL, Zou MB, Chen DX, Huang Y, Zhao
SL (2017) Unique approach to develop carbon dot-based
Page 9 of 9 185
nanohybrid near-infrared ratiometric fluorescent sensor for the de-
tection of mercury ions. Anal Chem 89:8044–8049
27. Yang JY, Yang T, Wang XY, Chen ML, Yu YL, Wang JH (2018)
Mercury speciation with fluorescent gold nanocluster as a probe.
Anal Chem 90:6945–6951
28. Du JJ, Yin SY, Jiang L, Ma B, Chen XD (2013) A colorimetric logic
gate based on free gold nanoparticles and the coordination strategy
between melamine and mercury ions. Chem Commun 49:4196–
4198
29. Zhao N, Lam J, Sung H, Su HM, Williams I, Wong KS, Tang BZ
(2014) Effect of the counterion on light emission: a displacement
strategy to change the emission behaviour from aggregation-caused
quenching to aggregation-induced emission and to construct sensi-
tive fluorescent sensors for Hg2+ detection. Chem Eur J 20:133–138
Publisher’s note Springer Nature remains neutral with regard to jurisdic-
tional claims in published maps and institutional affiliations.