Analytica Chimica Acta 947 (2016) 42e49

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Analytica Chimica Acta

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

Uncapped nanobranch-based CuS clews used as an efficient

peroxidase mimic enable the visual detection of hydrogen peroxide
and glucose with fast response
Xiangheng Niu a, b, *, Yanfang He b, Jianming Pan b, **, Xin Li b, Fengxian Qiu b,
Yongsheng Yan a, Libo Shi c, Hongli Zhao c, Minbo Lan c, ***
a
Institute of Green Chemistry and Chemical Technology, Jiangsu University, Zhenjiang 212013, PR China
b
School of Chemistry and Chemical Engineering, Jiangsu University, Zhenjiang 212013, PR China
c
Shanghai Key Laboratory of Functional Materials Chemistry, School of Chemistry and Molecular Engineering, East China University of Science and
Technology, Shanghai 200237, PR China

h i g h l i g h t s g r a p h i c a l a b s t r a c t

Uncapped nanobranch-based CuS

clews with large active surface and
abundant sites.

Peroxidase-like activity, with prefer-
able kinetics compared to HRP.

Outstanding performance for the
colorimetric sensing of H2O2.

Reliable monitoring of glucose in
clinical samples when in couple with
GOD.

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

Article history: Nanosized materials acting as substitutes of natural enzymes are currently attracting significant research
Received 30 July 2016 due to their stable enzyme-like characteristics, but some flaws of these nanozymes, including their
Received in revised form limited catalytic rate and efficiency, need to be remedied to enable their wider applications. In this work,
26 September 2016
we verify for the first time the catalytic behavior of uncapped nanobranch-based CuS clews as a
Accepted 3 October 2016
Available online 14 October 2016
peroxidase mimic. XRD, XPS, SEM, and TEM proofs demonstrate that high-purity CuS clews composed of
intertwined wires with abundant nanodendrites outside are successfully produced via a facile one-pot
hydrothermal synthesis approach, with thiourea as both the sulfion source and the structure-directing
Keywords:
Uncapped nanobranch-based CuS clews
agent. The synthesized CuS can catalytically oxidize 3,30 ,5,50 -tetramethylbenzidine (TMB) by H2O2 to
Mimetic peroxidase trigger a visible color reaction with rapid response (reaching a maximum change within 5 min). The
Colorimetric detection proposed CuS nanozyme exhibits preferable catalytic kinetics over natural horseradish peroxidase (HRP).
H2O2 This outstanding activity primarily results from the large surface area and rich sites exposed by the
Glucose uncapped unique structure. Under optimized conditions, the fabricated sensing system provides linear
absorbance (652 nm) changes in the H2O2 concentration range of 0.2e130 mM, with a detection limit of as
low as 63 nM. When coupled with glucose oxidase (GOD), the system is demonstrated to be capable of
monitoring glucose in blood samples with excellent performance.
© 2016 Elsevier B.V. All rights reserved.

* Corresponding author. Institute of Green Chemistry and Chemical Technology, Jiangsu University, Zhenjiang 212013, PR China.
** Corresponding author.
*** Corresponding author.
E-mail addresses: niuxiangheng@126.com (X. Niu), pjm@ujs.edu.cn (J. Pan), minbolan@ecust.edu.cn (M. Lan).

http://dx.doi.org/10.1016/j.aca.2016.10.013
0003-2670/© 2016 Elsevier B.V. All rights reserved.
 X. Niu et al. / Analytica Chimica Acta 947 (2016) 42e49
1. Introduction
Since Gao and co-workers first reported in 2007 that magnetic
Fe3O4 nanoparticles possessed intrinsic peroxidase-like activity
similar to natural horseradish peroxidase (HRP) [1], research has
been pursued to develop artificial nanomaterials as potential
peroxidase mimics [2e4]. Compared with natural enzymes, these
nanozymes are superior in several ways: 1) unlike bio-enzymes
that are much sensitive to environmental conditions including pH
and temperature, nanozymes are robust in harsh environments,
which gives them high stability and a long service life; 2) compared
to natural enzymes that are usually bio-proteins separated from
organisms, nanostructured materials used as mimetic enzymes are
relatively low-cost, which allows for their facile mass production
and large-scale use; 3) the catalytic behavior of these nanozymes
can be easily tuned, especially in combination with advanced
preparation and modification strategies; 4) some nanozymes, such
as Fe3O4, can be recycled for reuse. Given these properties,
nanomaterial-based artificial enzymes have attracted increasing
attention and interest from the analytical [5e11], environmental
[12e16], and biological [17e21] fields in recent years. In addition to
Fe3O4, attention has been focused on developing new materials as
effective enzyme mimics. Various materials including Co3O4 [7],
noble metals [6,22e24], Cu-based nanostructures [25e28], Fe-Co
bimetallic alloys [29], ITO nanocubes [30], graphitic C3N4 nano-
sheets [31], carboxyl-modified graphene oxide [32], reduced gra-
phene oxide [13], Si dots [33], and Prussian blue [34] have been
demonstrated to be capable of catalyzing H2O2 to generate hy-
droxyl radicals. Among these materials, CuS has drawn special
attention in the nanozyme field. Several CuS structures with
interesting shapes have been explored for their sensing application
[26e28]. Furthermore, versatile preparation methods and param-
eters can result in the diversity of the morphology, composition and
valence state, and microstructure of CuS. Therefore, the peroxidase-
like catalytic property of CuS structures is easy to be tuned by
adjusting these characters.
Despite the above characteristics of nanozymes, they still have
some drawbacks that need to be addressed to enable their broad-
scale use. One of these flaws in nanozymes is their limited cata-
lytic efficiency [2]. The poor activity of nanozymes is primarily due
to the following reasons: 1) unlike natural enzymes and organic
catalysts that are usually in a homogeneous phase, nanostructure-
based enzyme mimics undergo a heterocatalysis process, in which
the contact probability of zymolyte-enzyme, electron transfer, and
mass diffusion are significantly reduced; 2) the catalytic behavior of
nanozymes is highly dependent on their shape, size, composition,
microstructure, etc. [1,2]; 3) the surface of a nanostructure is usu-
ally coated with some capping agents that are utilized in the syn-
thesis process [35], and thus some active sites will be blocked from
reacting. As the catalytic activity of current nanozymes primarily
originates from active sites on the material's surface, simulta-
neously exposing sufficient active sites to substrates by enlarging
the available surface of nanozymes and avoiding surface coating
may be an efficient approach to improve their catalytic efficiency.
Given these considerations, here we demonstrate for the first
time the impressive catalytic efficiency of new CuS clews as a
mimetic peroxidase. The high-purity CuS clews are composed of
intertwined wires, with abundant nanodendrites grown around
individual wire and no capping agent coated on their surface, thus
providing a large surface area and abundant reaction sites. As a
result, the synthesized CuS structure is able to catalytically oxidize
3,30 ,5,50 -tetramethylbenzidine (TMB) in the presence of H2O2 to
produce a visible color reaction, with preferable catalytic kinetics
over natural horseradish peroxidase (HRP). The wide linear range
43
and low detection limit of H2O2 analysis associated with the pro-
posed sensing system are demonstrated. The performance of the
fabricated system in combination with glucose oxidase (GOD) for
glucose monitoring in clinical samples is also investigated.
2. Materials and methods
2.1. Chemicals
Thiourea, CuSO4$5H2O, and H2O2 (30% aqueous, v/v) were
received from Sinopharm Chemical Reagent Co., Ltd. TMB, ascorbic
acid (AA), dopamine (DA), 4-acetaminophen (AP), glucose, fructose,
and ribose were supported by Shanghai Aladdin Reagent Co. HRP
(3500 units/mg) was provided by Shanghai Macklin Biochemical
Co., and GOD (from Aspergillus niger, 200 units/mg) was purchased
from Sigma-Aldrich. Ultrapure water was used throughout the
study. All other reagents were at least of analytical grade, and used
directly without further purification.
2.2. Synthesis of uncapped nanobranch-based CuS clews
CuS clews were prepared via a facile one-pot hydrothermal
synthesis approach. 0.15 g thiourea and 0.5 g CuSO4$5H2O were
first dissolved in 30 mL ultrapure water; the precursor solution was
then sealed in a Teflon-lined stainless steel reactor, followed by
heat treatment at 180  C for 2 h. Afterwards, the products were
collected by centrifugation and washed with water, and finally
dried in a vacuum oven at 40  C for 24 h.
2.3. Characterization
The crystal structure of the as-synthesized products was
determined by an X-ray diffractometer (XRD, XRD-6100Lab, Shi-
madzu) with Cu Ka radiation. The surface chemistry of the mate-
rials was probed by an X-ray photoelectron spectrometer (XPS,
ESCALAB 250Xi, Thermo Fisher). Scanning electron microscopy
(SEM, JSM-6360LV, JEOL) was used to observe the morphology of
the products. Transmission electron microscopy (TEM) images
were captured on a JSM-2100 microscope (JEOL) with an acceler-
ation voltage of 100 kV. The CuS concentration in solutions was
deduced by accurately detecting the Cu content with inductively
coupled plasma optical emission spectroscopy (ICP-OES, IRIS-1000,
Thermo Fisher).
2.4. Colorimetric measurements
All colorimetric measurements were made on a 722s visible
spectrophotometer (Lengquang Tech. Co., China). Time-dependent
absorbance changes were recorded by mixing a specific amount
of TMB, H2O2, and enzyme (nanosized CuS or HRP) into a 0.2 M
NaAc-HAc buffer solution and immediately measuring the mixture.
Reaction kinetics measurements were performed by recording the
absorbance value at 652 nm with a 1 min interval, and the apparent
kinetics parameters were calculated using the Michaelis-Menten
equation.
For the colorimetric analysis of H2O2, targets with various con-
centrations were added into the NaAc-HAc buffer solution under
optimized conditions. After undergoing the reaction for 5 min, the
solution was ready for colorimetric measurements.
Glucose detection was performed as follows: 1) 50 mL of
1 mg mL1 GOD and glucose at various levels in 1 mL of 0.1 M
phosphate buffer (pH 7.4) were incubated at 37  C in a water bath
for 15 min; 2) optimized amounts of TMB and CuS nanozyme and
4 mL of 0.2 M NaAc-HAc buffer (pH 4.0) were successively added
44 X. Niu et al. / Analytica Chimica Acta 947 (2016) 42e49
into the above solution; 3) the final mixture was further incubated
for 5 min, and then the absorbance value at 652 nm was recorded.
3. Results and discussion
3.1. Characterization of uncapped nanobranch-based CuS clews
An advantage of nanozymes compared to natural enzymes is
their simple preparation and modification, which makes their mass
production and large-scale application possible. In this work,
nanosized CuS clews are prepared via a facile one-pot hydrother-
mal synthesis method, in which CuSO4$5H2O is the Cu2þ precursor
and thiourea is both the sulfion source and the structure-directing
agent. The crystalline phase of the as-prepared products is deter-
mined by XRD. As shown in Fig. 1(A), all diffraction peaks can be
indexed to a pure hexagonal phase of CuS (JCPDS No. 06-0464). No
other impurity is detected, demonstrating the high purity of CuS.
Additionally, the sharp peaks indicate that the products are well
crystallized. The surface chemistry of the as-synthesized CuS is
probed by XPS. As depicted in Fig. 1(B), the S and Cu elements are
found, with no impure component detected (the C and O elements
may originate from the residual air during XPS measurement).
Fig. S1 (Supporting Information) presents the high-resolution XPS
spectrum of Cu2p. Well-defined peaks attributed to the CuII species
are observed. Both the XRD and XPS measurements verify that the
surface of the synthesized CuS is pure, with no agent coated on its
surface. As a matter of fact, no capping agent is added into the
preparation system, and the NH3 molecule decomposed from
thiourea is prone to desorption from the CuS surface [36]. SEM and
TEM are used to observe the shape and microstructure of the as-
prepared CuS. As shown in Fig. 2(A), a clew-like CuS structure is
obtained. The CuS clews are composed of some intertwined wires,
and the surface of individual wire seems to be quite rough. This
feature is also confirmed by the TEM images shown in Fig. 2(B).
Many short nanodendrites grow outside each CuS wire. This
structure of uncapped CuS may favor heterocatalysis, because it is
supposed to offer a large surface area and many active sites for
substrates. Fig. 2(C) shows the high-resolution TEM image of
uncapped CuS. The lattice distance of 0.30 nm reveals that the (102)
crystal face of nanosized CuS is well exposed. In addition, the
uncapped CuS nanostructures show good dispersibility in the
NaAc-HAc buffer solution, with no obvious sedimentation in a short
period, as demonstrated by Fig. S2(A) (Supporting Information).
After a long time (3 days), some sediments are observed at the tube
bottom, as shown in Fig. S2(B) (Supporting Information). After
simple shake, the deposited CuS nanostructures are easily re-
dispersed in the buffer solution again.
Fig. 1. XRD (A) and XPS (B) patterns of uncapped nanobranch-based CuS clews.
3.2. Rapidly catalytic oxidation of TMB by nanosized CuS in the
presence of H2O2
The peroxidase-like activity of uncapped nanobranch-based CuS
clews is investigated using TMB as a typical peroxidase substrate in
the presence of H2O2. For comparison, another three parallel sys-
tems with different combinations of TMB, H2O2, and nanosized CuS
are also checked. Fig. 3 shows the time-dependent absorbance
(652 nm) changes of the four reaction systems. In the A, B, and C
systems, a negligible color change is found, as the absorbance value
has no obvious increase, indicating that the three systems (without
nanozyme in A, without TMB in B, and without H2O2 in C) can
hardly induce the color reaction. In contrast, a visible color change
of TMB is observed when TMB, H2O2, and nanosized CuS are
simultaneously added into the reaction system (D). The colorless
buffer solution turns blue-green, with a maximum absorption
wavelength of 652 nm [23]. This color change is attributed to the
oxidation of TMB into TMBox [37]. The related reaction process has
been described elsewhere [5,13,23]. Typically, the OeO bond of
H2O2 adsorbed on the surface of nanosized CuS will first be broken
up into two hydroxyl radicals catalyzed by the CuS peroxidase
mimic, and then the generated hydroxyl radicals will trigger the
oxidation of colorless TMB into blue-green TMBox. This result
demonstrates that the synthesized CuS exhibits peroxidase-like
characteristics.
Excitingly, the catalytic oxidation of TMB by H2O2 in the pres-
ence of nanosized CuS shows a fast reaction rate, as the absorbance
(652 nm) reaches to its maximum value within 5 min, which seems
to be much quicker than Co3O4-reduced graphene oxide [7] and Au
NPs supported on MoS2 NRs [5]. This phenomenon indicates that
the system undergoes a rapid reaction process. As the generation of
hydroxyl radicals from H2O2 catalyzed by nanozymes dominates
the total reaction rate, the fast color change reveals that the syn-
thesized CuS has high activity as a mimetic peroxidase. In addition,
it is found that the catalytic activity of nanosized CuS is dependent
on the reaction conditions, including the pH and the concentrations
of TMB, H2O2, and CuS. The relationships between the peroxidase
activity of uncapped nanobranch-based CuS clews and the reaction
parameters are displayed in Fig. S3 (Supporting Information). As
shown, a pH value of e4, 40 mg/mL CuS, 0.2 mM TMB and
0.35 M H2O2 are the optimal reaction conditions.
3.3. Kinetics analysis of the synthesized CuS as a peroxidase mimic
The outstanding activity of uncapped nanobranch-based CuS
clews as a peroxidase mimic is further confirmed by standard
steady-state kinetics measurements. For comparison, HRP, a natural
 X. Niu et al. / Analytica Chimica Acta 947 (2016) 42e49
Fig. 2. SEM (A) and TEM (B and C) images of uncapped nanobranch-based CuS clews.
Fig. 3. Time-dependent absorbance (652 nm) changes of TMB in different reaction
systems in 0.2 M NaAc-HAc buffer (pH 4): (A) 500 mM H2O2 þ 0.125 mM TMB; (B)
500 mM H2O2 þ 40 mg/mL nanosized CuS; (C) 0.125 mM TMB þ 40 mg/mL nanosized
CuS; (D) 500 mM H2O2 þ 0.125 mM TMB þ 40 mg/mL nanosized CuS. The inset shows
the color reaction images of different reaction systems. (For interpretation of the ref-
erences to colour in this figure legend, the reader is referred to the web version of this
article.)
peroxidase, is also tested under these conditions. A series of ex-
periments are performed by varying the concentration of one
substrate (TMB or H2O2) and keeping the other substrate (H2O2 or
45
TMB) concentration constant. In a certain concentration range of
H2O2 or TMB, typical Michaelis-Menten curves are observed, as
depicted in Fig. 4(A) and (B) for TMB and H2O2, respectively. The
data are fitted according to the following Lineweaver-Burk
equation:
    
1 Km 1 1
¼ þ (1)
n Vmax ½S Vmax
where n is the initial rate, Km is the Michaelis-Menten constant,
Vmax is the maximal reaction rate, and [S] is the substrate concen-
tration. The calculated Km and Vmax are listed in Table 1. Km is an
indicator of the affinity of an enzyme toward a substrate, and the
smaller the value of Km, the stronger the affinity between the
enzyme and the substrate. The synthesized CuS exhibits a stronger
affinity toward TMB than toward H2O2, in consistent with the fact
that a higher H2O2 level than TMB is required to achieve the
maximum activity. Similar results are also found in the case of HRP.
The Km value of HRP toward TMB measured under standard con-
ditions ise0.4 mM, similar to the value in the previous report [1].
However, the Km value toward TMB obtained on the synthesized
CuS is calculated to bee0.06 mM, much lower than that of HRP. This
result indicates that the proposed CuS clews have a higher affinity
toward TMB than HRP. A similar phenomenon is also observed with
H2O2 as the substrate. In terms of Vmax, the values measured on
nanosized CuS are much higher than the corresponding cases on
HRP, suggesting the higher peroxidase activity of the former.
All of these results suggest that the synthesized CuS is able to
exhibit catalytic behavior when it is used as a mimetic peroxidase.
The lower Km values demonstrate stronger affinity toward sub-
strates, and the larger Vmax implies the faster response. The
46 X. Niu et al. / Analytica Chimica Acta 947 (2016) 42e49
Fig. 4. Steady-state kinetics measurements of nanosized CuS under the optimized pH and nanozyme concentration. (A) The concentration of H2O2 was 0.35 M and the TMB
concentration varied. (B) The concentration of TMB was 0.2 mM and the H2O2 concentration varied.
Table 1
Comparison of the kinetics parameters of nanosized CuS and HRP.
Catalyst Substrate Vmax [108 M s1] Km (mM)
HRP TMB 10.8 0.379
HRP H2O2 9.7 4.531
Nanosized CuS TMB 76.4 0.064
Nanosized CuS H2O2 23.7 1.753
surprising activity of the synthesized CuS nanozyme may originate
from several superiorities: 1) compared to HRP with only one iron
ion as the active center, nanosized CuS has many active sites on the
nanozyme surface; 2) the synthesized clew-like CuS is composed of
many intertwined wires, and around each wire abundant short
nanodendrites are formed. This structure certainly offers large
surface and sufficient sites for enzyme-substrate contact for further
reaction; 3) as the synthesized CuS is quite pure in composition,
with no adsorbed agent capped on its surface, thus exposing all
active sites for the catalytic reaction.
3.4. Robustness comparison of the synthesized CuS nanozyme and
HRP
Another benefit of nanozymes compared to natural enzymes is
their robustness and stability against reaction and storage condi-
tions. To demonstrate this assumption, we measure the catalytic
activity of both nanosized CuS and HRP after incubation at a range
of temperatures and in solutions with different pH values. It is
found that the activity of the HRP enzyme decreases when the
incubation temperature is higher than 45  C, while the proposed
CuS nanozyme maintains stable catalytic activity over a wide
temperature range from 25 to 95  C, as shown in Fig. 5(A). In so-
lutions with different pH values, the maximum activity of HRP is
obtained in neutral media. With pH decreases or increases, the HRP
activity is significantly reduced, with no activity in strong acid or
base. In contrast, the CuS nanozyme exhibits no significant change
in activity when incubated in a wide range of pH from 1 to 13, as
depicted in Fig. 5(B). These measured data confirm the higher
temperature and pH resistance of nanosized CuS compared to
natural HRP. Additionally, the synthesized CuS exhibits outstanding
long-term stability, with no significant change of its catalytic ac-
tivity after 10 days of storage in air at 40  C. The robustness of the
prepared nanozyme makes it quite suitable for a broad range of
applications.
3.5. Colorimetric analysis of H2O2 and glucose
As the absorbance (652 nm) changes are dependent on the H2O2
levels, the reaction system can be utilized for the quantitative
determination of the substrate H2O2, which is of significant
research interest in various fields [38e40]. Under optimized con-
ditions, the absorbance (652 nm) values of the system after reaction
for 5 min are recorded when H2O2 with various concentrations is
added. As shown in Fig. 6(A), a linear increase of the absorbance
value is found when the H2O2 content varies from 0.2 to 130 mM,
with a detection limit (LOD) of as low as 63 nM, which is calculated
based on the signal-to-noise (S/N) of three protocol. When more
H2O2 is added into the sensing system, a common saturation effect
is observed. We also compare the degrees of color change induced
by different H2O2 concentrations (top-left inset), and the color
differences can be visually identified, indicating the visual deter-
mination of the trace target. Compared with other similar nano-
zymes, as listed in Table S1 (Supporting Information), the proposed
CuS provides superior performance in term of the fast response,
wide linear scope, and low LOD. These excellent characteristics of
nanosized CuS primarily result from its high catalytic activity and
efficiency, which are due to its large active surface and abundant
reaction sites.
Additionally, the fabricated sensing system has good reproduc-
ibility, with a relative standard deviation (RSD) of 2.3% for eight
measurements of 5 mM H2O2. With respect to the detection selec-
tivity, the possible interference from several common species
including AA, DA, AP, and glucose is assessed. As depicted in
Fig. 6(B), 0.1 mM AA, DA and AP, and 1 mM glucose have no obvious
interference for the selective determination of 100 mM H2O2. The
recovery test results, as listed in Table S2 (Supporting Information),
suggest that the fabricated sensing system can be used to detect the
target with high reliability.
As the generation and consumption of H2O2 are related to
various chemical and biological processes, various species and re-
actions in which H2O2 is involved can be theoretically probed using
the proposed colorimetric system [5,9,10,17,21,22]. Here we verify
the feasibility of the system for glucose sensing. As illustrated by
Fig. 7(A), the added GOD catalytically oxidizes glucose into gluconic
acid, simultaneously producing H2O2. By determining the amount
of H2O2 produced, the glucose target can also be quantitatively
detected. The absorbance (652 nm) increases linearly with
increasing glucose amounts in the scope of 0.5e110 mM, as shown in
Fig. 7(B), with apparent color changes that can be visually identified
(top-left inset). The LOD of glucose detection is further calculated to
be as low as 0.13 mM based on S/N ¼ 3. The detection range and LOD
 X. Niu et al. / Analytica Chimica Acta 947 (2016) 42e49 47

Fig. 5. Comparison of the catalytic stability of nanosized CuS and HRP. (A) Nanosized CuS and HRP were first incubated in buffer solutions at various temperatures for 0.5 h and then
their activities were measured under optimized conditions, and the activity measured at room temperature (25  C) was set as 100%. (B) Nanosized CuS and HRP were first incubated
in buffer solutions with various pH values for 0.5 h and then their activities were measured under optimized conditions, and the maximum point in each curve was set as 100%.

Fig. 6. (A) The relationship between the recorded absorbance (652 nm) and the H2O2 concentration. The inset in top left displays the color reaction images of different H2O2
concentrations. (B) The absorbance (652 nm) values caused by different species (100 mM H2O2, AA, DA and AP, and 1 mM glucose). (For interpretation of the references to colour in
this figure legend, the reader is referred to the web version of this article.)

of the fabricated system for glucose detection are comparable to or
even better than similar methods, as summarized in Table S3
(Supporting Information). Eight independent tests lead to a calcu-
lated RSD of 3.1%, indicating the repeatability of the colorimetric
method.
The selectivity of the proposed system is also evaluated. The
absorbance responses of another two sugars, fructose and ribose,
and AA and AP are recorded and compared with that of glucose
with the same concentration (0.1 mM). As the excellent specificity
of GOD toward glucose and the peroxidase-like feature of nano-
sized CuS, a negligible change in the absorbance is found when
these species are added into the reaction system instead of glucose,

Fig. 7. (A) Illustration of the mechanism of detecting glucose with the combination of GOD and nanosized CuS. (B) The relationship between the recorded absorbance (652 nm) and
the glucose concentration, and the inset in top left displays the color reaction images of different glucose concentrations. The inset in bottom right exhibits the absorbance (652 nm)
values caused by different species with the same concentration (0.1 mM). (For interpretation of the references to colour in this figure legend, the reader is referred to the web
version of this article.)
48 X. Niu et al. / Analytica Chimica Acta 947 (2016) 42e49
Table 2
Comparison of the results of glucose analysis in clinical samples detected by the
standard glucometer and the method proposed in this work.
Samplea Sannuo glucometer (mM)b Proposed method (mM ± SD, n ¼ 3)c
1# 8.9 8.87 ± 0.13
2# 7.3 7.41 ± 0.09
3# 10.4 10.33 ± 0.21
a
These blood samples were provided by the Jiangbin Hospital of Zhenjiang.
b and its application in visual biosensing of glucose, Anal. Chim. Acta 796 (2013)
The glucose detection was performed directly (without any dilution) with the
Sannuo glucometer.
c [8] H. Wei, E.K. Wang, Fe3O4 magnetic nanoparticles as peroxidase mimetics and
The blood samples were diluted 100-folds for glucose detection by the proposed
method.
demonstrating its outstanding selectivity for glucose detection. The
proposed method is further used to monitor glucose in clinical
samples. A commercial Sannuo glucometer (Sinocare Inc., China) is
used as the standard tool for comparison. The obtained results are
listed in Table 2. The proposed method is verified with good ac-
curacy and reliability for the analysis of glucose in serum.
4. Conclusions
In summary, we have introduced a new mimetic peroxidase,
uncapped nanobranch-based CuS clews, with outstanding catalytic
activity and efficiency for the visual sensing of H2O2 and glucose.
The CuS nanozyme obtained by a one-pot hydrothermal synthesis
has a large active surface area and abundant reaction sites, with
superior Michaelis-Menten kinetics to natural HRP. With their
amazing and stable peroxidase-like activity, the developed sensing
systems provide excellent performance for H2O2 and glucose
detection, with a wide linear range and a low LOD. The nanosized
CuS acts as a peroxidase mimic, with the benefits of easy synthesis,
low cost, and excellent activity as well as high stability under harsh
conditions. These properties will allow for wider applications in the
environmental, biological, and analytical fields. We are currently
exploring ways to further improve the peroxidase-like efficiency of
nanosized CuS for more challenging applications, e.g., the direct
monitoring of trace superoxide anion in biological fluids.
Acknowledgments
This work was financially supported by the National Natural
Science Foundation of China (Nos. 21605061 and 31601549), the
Natural Science Foundation of Jiangsu Province (No. BK20160489),
the Natural Science Fund for Colleges and Universities in Jiangsu
Province (No. 16KJB150009), the Open Fund from Shanghai Key
Laboratory of Functional Materials Chemistry (No. SKLFMC201601),
and the Start-Up Research Fund of Jiangsu University (No.
15JDG143).
Appendix A. Supplementary data
Supplementary data related to this article can be found at http://
dx.doi.org/10.1016/j.aca.2016.10.013.
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