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Colorimetric Detection of Sulte in Foods by a TMBO2Co3O4

Nanoparticles Detection System
Wenjie Qin,, Li Su, Chen Yang,, Yanhua Ma,, Haijuan Zhang,, and Xingguo Chen*,,

State Key Laboratory of Applied Organic Chemistry, Lanzhou University, Lanzhou 730000, China

Department of Chemistry, Lanzhou University, Lanzhou 730000, China

School of Chemistry and Chemical Engineering, Henan Normal University, Xinxiang 453007, China
S Supporting Information

ABSTRACT: In this paper, we rst discovered that Co3O4 nanoparticles (NPs) possess intrinsic oxidase-like activity and can
catalytically oxidize peroxidase substrates, such as 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt
(ABTS) and 3,3,5,5-tetramethylbenzidine (TMB), to form colored products, in the absence of exogenously added H2O2. The
presence of sulte inhibited the TMBO2Co3O4 NPs reaction system and caused a change in color of the reaction system. On
the basis of this phenomenon, a colormetric approach to detect sulte was established with a good linear relationship ranging
from 0.2 106 to 1.6 105 M and a detection limit of 5.3 108 M. The method was used to detect sulte in foods. Good
recoveries ranging from 93.8% to 100.5% were obtained. Furthermore, the mechanism was studied and results showed that the
oxidase-like activity of the Co3O4 NPs was not from OH or O2 radical generated. It may probably originate from their ability
to transfer an electron between the peroxidase substrate and oxygen absorbed on the surface of the Co3O4 NPs.
KEYWORDS: Co3O4 nanoparticles, oxidase mimic, colorimetry, sulte, mechanism of oxidase-like activity

Sulte is one of the oldest and most ubiquitous food additives,
nanomaterials,18,19 metal nanomaterials,13,20 carbon nanoma-
terials,13,20 nanocomposites,2128 and other nanomaterials.2931
These enzyme-like nanomaterials could be employed in
which has been widely used as blanching and preservative
colorimetry to detect nucleotides,32 H2O2,3335 glucose,3335
agents in a large variety of foodstus (dried fruit, vermicelli,
melamine,36 and so on. To the best of our knowledge, however,
preserves, etc.) to improve the appearance of food and prevent
the colorimetric detection of sulte in food using the enzyme-
bacterial growth.1,2 However, the ingestion of foods containing like property of nanoparticles has not been explored up to now,
large amounts of sulte can cause asthmatic attacks and allergic which might be attributed to the complication of food samples
reactions in sensitive individuals.3,4 Owing to the potential matrices and the diculties in constructing an eective
toxicity, the sulte in foods is strictly limited in many nanoparticles-based enzyme mimic sensing platform to detect
countries.5 Hence, sulte determination in food products is micro amounts of food additive.
essential for the purposes of public health, food assurance, and Herein, we have developed a TMBO2Co3O4 NPs
quality control. In recent years, numerous methods have been colorimetric system for detection of sulte in foods, based on
developed for determining sulte in foods, including titrimetry, the property of oxidase-like activity of Co3O4 NPs. In this
ow injection analysis, anodic stripping voltammetry, ion method, the existence of sulte can cause a change in color of
chromatography, chemiluminescence, gas chromatography, the reaction system, and the color change can be visually
uorometry, high-performance liquid chromatography, and observed directly. We also investigated the mechanism of
colorimetry.68 oxidase-like activity of Co3O4 NPs by XPS, contrast experi-
Colorimetric biosensing has become more attractive because ments, and cyclic voltammetry.

of its low cost, simplicity, and practicality. However, the key
challenge for colorimetric sensors is transforming the detection EXPERIMENTAL PROCEDURES
events into color changes. Since color changes can be read by
Materials. Co(CH3COO)24H2O were obtained from Tianjin
the naked eye, a colorimetric sensor does not require expensive Guangfu Chemical Reagent Factory (Tianjin, China). 3,3,5,5-
or sophisticated instrumentation and can be applied to eld Tetramethylbenzidine (TMB) and 2,2-azinobis(3-ethylbenzothiazo-
analysis.911 To this end, enzyme-like nanomaterials-based line-6-sulfonic acid) diammonium salt (ABTS) were purchased from
sensors have emerged as an important colorimetric tool. Since Sigma-Aldrich. Ammonium hydroxide (2528 wt %) was purchased
the rst report of Fe3O4 magnetic nanoparticles (MNPs) from Baiyin Liangyou Chemical Reagent Factory (Baiyin, China).
possessing intrinsic peroxidase-like activity,12 nanoparticles- Sodium acetate, glacial acetic acid, anhydrous sodium sulte, sodium
based enzyme mimics have received great attention due to their
merits relative to native enzymes (e.g., low cost, high stability, Received: February 23, 2014
and tunability in catalytic activity). More and more nano- Revised: May 28, 2014
particles have been found as enzymatic mimics, such as metal Accepted: June 2, 2014
oxide nanomaterials,13 sulde nanomaterials,1417 selenide

XXXX American Chemical Society A | J. Agric. Food Chem. XXXX, XXX, XXXXXX
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Figure 1. Images of the oxidation color reaction of TMB and ABTS by Co3O4 NPs (a). UV/vis spectra of TMB solution in 0.2 M acetic acidacetate
buer (pH 4.0) at 40 C for 10 min. The TMB and Co3O4 NPs concentrations were 0.25 mM and 40 g mL1, respectively (b). UV/vis spectra of
an ABTS solution in 0.2 M acetic acidacetate buer (pH 4.0) at 40 C for 10 min. The ABTS and Co3O4 NPs concentrations were 0.25 mM and
40 g mL1, respectively (c).

Figure 2. Survey XP spectra of Co3O4 NPs before and after reaction (a and d). Deconvoluted high-resolution Co 2p3/2 XP spectra of Co3O4 NPs
before and after reaction (b and e). Deconvoluted high-resolution O 1s XP spectra of Co3O4 NPs before and after reaction (c and f).

dihydrogen phosphate, and other regents were obtained from Tianjin D/Max-2400 X-ray diractometer with Cu K radiation ( =
Guangfu Chemical Reagent Factory (Tianjin, China). A sulte 0.154056). The chemical state of the coating surface was analyzed
solution was prepared daily by dissolving anhydrous sodium sulte using X-ray photoelectron spectroscopy (XPS, PerkinElmer PHI-5702
in water and was standardized by iodimetric titration when needed. All multifunctional photoelectron spectrometer). Element determination
other chemicals were of analytical reagent grade and used without was measured by a 4300 DV inductively coupled plasmaatomic
further purication. Deionized water was used throughout the emission spectrometer (ICP-AES). Zeta potential measurements of
experiment. Co3O4 NPs were performed with a Malvern Nano-ZS apparatus. ESR
Apparatus and Characterization. A TU-1901 double-beam spectra were acquired using a JES-FA200 electron spin resonance
UVvis spectrophotometer (Beijing Purkine General Instrument Co. spectrometer. Electrochemical experiments were performed on a CHI
Ltd., China) was used to record absorption spectra and measure 832 electrochemical workstation (Shanghai Chenhua Instrument Co.,
absorbance. A FEI Tecnai G2 F30 microscope (TEM) was used for China) and a conventional three-electrode system. The conventional
measuring the morphologies and size of Co3O4 NPs. The X-ray three-electrode system contained a Pt wire counter electrode, a
diraction (XRD) pattern of Co3O4 NPs was determined on a Rigaku saturated calomel electrode as reference electrode, and an as-prepared

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working electrode. The specic surface area was measured using a

multipoint BrunauerEmmettTeller technique (ASAP 2020, Micro-
meritics, USA).
Preparation of Co3O4 Nanoparticles. Co3O4 NPs were prepared
using a hydrothermal method as described in the literature.37 Briey,
0.5 g of Co(CH3COO)24H2O was dissolved in 25 mL of ethanol, and
then 2.5 mL of ammonia solution was added under vigorous stirring.
After stirring for 10 min, the obtained suspension was transferred into
a 50 mL autoclave. Then the autoclave was sealed and kept at 150 C
for 3 h followed by cooling to room temperature naturally. Finally, the
product was separated via centrifugation and alternately washed
thoroughly several times with deionized water and ethanol until the
pH of supernatant was about 7.0, then dried at 60 C for 4 h.
Kinetic Assay of Oxidase-like Co3O4 NPs. Kinetic experiments
were carried out at 40 C using 120 L of 1.0 mg mL1 Co3O4 NPs in
2.8 mL of acetic acidacetate buer solution (0.2 M, pH 4.0) with 80
L of TMB/ABTS as substrate, unless otherwise stated. Kinetic Figure 4. Cyclic voltammograms of bare GCE (a) and the Co3O4
measurements were carried out in time course mode by monitoring NPs-modied GCE (b) in a 0.2 M acetic acidacetate buer (pH 4.0)
the absorbance change at 652/417 nm for 4 min using a TU-1901 + 0.25 mM TMB solution.
UVvis spectrophotometer. The MichaelisMenten constant was
calculated using a LineweaverBurk plot:12
1/v = K m/Vmax(1/[S] + 1/K m) Similar to other peroxidase/oxidase mimic reactions, the
maximum absorbance peaks of the oxidation products of
where is the initial velocity, Vmax represents the maximal reaction TMB and ABTS were located at 652 and 417 nm,
velocity, and [S] is the substrate concentration. Km is the Michaelis respectively.12,33 Nevertheless, there was no absorbance at
Menten constant, which is an indicator of enzyme anity for its
substrate. 652 or 417 nm in the absence of Co3O4 NPs in the system
Preparation of Samples. Dried lily bulbs, vermicelli, and haw roll (Figure 1b and c). The results suggested that Co3O4 NPs
were purchased randomly from a local market in Lanzhou, China. exhibit intrinsic oxidase-like activity. The presence of mixed
Samples were minced by a mixer. An accurately weighed minced valence states of Co2+ and Co3+ ions in Co3O4 NPs is
sample was transferred to a beaker, and 40 mL of EDTA (1.0 104 considered the key in their catalysis. As an oxidase mimic, the
M) was added.5 After shaking and ultrasonication for 15 min, the states of Co in the Co3O4 NPs did not change before or after
mixture was ltered with a 0.45 m syringe membrane lter. The the reaction.
ltrate was diluted to 100 mL with EDTA (1.0 104 M). All samples In order to further conrm the property of oxidase mimic of
were stored in a refrigerator at 4 C before analysis. the Co3O4 NPs, XPS was performed to nd the oxidation state
Detection of Sulte. The amount of sulte in samples was
determined according to the following steps: (a) 80.0 L of sample of Co in the Co3O4 NPs before and after the reaction. Figure 2e
solution, 150.0 L of 5 mM TMB, and 120.0 L of Co3O4 NPs shows the deconvoluted XPS spectra of Co 2p3/2 of Co3O4 NPs
solution (1.0 mg mL1) were added into 2.65 mL of a 0.2 M acetic after the reaction. The relative ratio of Co2+ and Co3+ was
acidacetate buer solution (pH 4.0) and then thoroughly mixed to almost unchanged in Co3O4 NPs before and after the reaction.
make it uniform; (b) the as-mixed solution was maintained in a 40 C The high-resolution XPS spectra O 1s of unused and used
water bath for 10 min to hold the reaction and then kept in an ice Co3O4 NPs are presented in Figure 2c and f, respectively. The
water bath for 10 min to terminate the reaction. The solution was then relative ratio of OCo2+ and OCo3+ peaks in O 1s XPS
used to perform the adsorption spectroscopy measurement at 652 nm spectra was similar to Co 2p3/2 XPS results (Table S1). It is
wavelength. For the control experiments, 80.0 L of deionized water suggested that the states of Co in the Co3O4 NPs did not
was used instead of sample solution.

change before or after the reaction. These results conrmed

that Co3O4 NPs exhibited intrinsic oxidase-like activity.
It is important to conrm the possibility that the observed
Oxidase-like Activity of Co3O4 NPs. The oxidase-like activity was not caused by cobalt ions leaching from Co3O4 NPs
activity of Co3O4 NPs was evaluated in the catalysis of typical in acidic solution. To test this, Co3O4 NPs were incubated in
substrates TMB and ABTS in the absence of H2O2. As can be the reaction buer (pH 4.0) for 2 h and then removed from the
seen from Figure 1a, Co3O4 NPs catalyzed the oxidation of solution by centrifugation. As shown in Figure S5, comparing
TMB and ABTS to produce the typical colored products. the activity of the leaching solution with that of Co3O4 NPs
under the same conditions, the leaching solution had no
activity, showing that the observed oxidase-like activity was due
to intact Co3O4 NPs. Figure S6 shows the amount of cobalt
ions leaked from Co3O4 NPs in the buer solutions (pH 3.0
7.0) for 2 h, which was implemented by ICP-OES. The ICP
results exhibited that there were few cobalt ions in the leaching
solution. They probably came from residual Co3O4 NPs after
Figure 3. Photographs of TMB solutions in the absence and presence
Possible Mechanism of Oxidase-like Activity of Co3O4
of an O2 oxidation reaction catalyzed by Co3O4 NPs (a). Photographs NPs. As an oxidase mimic, Co3O4 NPs catalyze an oxidation
of a TMB solution oxidation reaction catalyzed by Co3O4 NPs without reduction reaction involving oxygen (O2) as the electron
degassing and Co3O4 NPs degassed by three freezepumpthaw acceptor. However, in the reaction system of TMBO2Co3O4
cycles (b). Photographs of the oxidation color reaction of TMB, TMB- NPs, oxygen is present in the aqueous solution and also exists
SO32, and TMB-SO42 catalyzed by Co3O4 NPs, respectively (c). on the surface of Co3O4 NPs due to the high specic surface
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Scheme 1. Possible Mechanism for the TMBO2Co3O4 NPs System

Figure 5. MichaelisMenten curve of Co3O4 NPs with TMB (a). MichaelisMenten curve of Co3O4 NPs with ABTS (b). Details are included in
the Experimental Procedures.

area of nanomaterials. In order to determine whether the bond strength of Co3O4 and the larger surface energy of
oxygen in the solution played a role in the TMBO2Co3O4 nanomaterials caused by a large surface to volume ratio.39 In
NPs system, two sets of experiments were conducted: the rst addition, the O 1s peak at 533.0 eV in the XPS spectrum shown
one was carried out in the presence of oxygen; the other was in Figure 2c indicated that there was chemisorbed oxygen on
performed under high-purity nitrogen and the solution was the surface of Co3O4 NPs.40 It is indicated that both the
degassed before the reaction, as in the photograph shown in physically absorbed and chemisorbed oxygen on the surface of
Figure 3a. It was found that they exhibited the same color. The Co3O4 NPs played a key role in the TMBO2Co3O4 NPs
absorbance of the rst one was 0.747; the other was 0.770 at system. In order to further conrm the above conclusion,
652 nm. The absorbance of the latter was higher than the oxidation experiments of TMB, TMB-SO32, and TMB-SO42
former; one possible reason was that nitrogen played a role in catalyzed by Co3O4 NPs were carried out. The results are
mixing and speeded up the reaction. This indicated that the shown in Figure 3c. TMB-SO32 system presented a lower
oxygen in the solution had no eect on the reaction, which was absorbance than the other two systems. Since the volume of
dierent from the behavior of some nanoparticles-based oxidase SO32 is smaller than TMB, and Co3O4 NPs have a positive
mimics reported.8,38 For further study, the Co3O4 NPs without charge at pH 4.0 (Supporting Information), SO32 is more
degassing and Co3O4 NPs degassed by three freezepump easily adsorbed on the surface of Co3O4 NPs. Then SO32
thaw cycles that catalyze the oxidation of TMB were consumed oxygen absorbed on the surface of the Co3O4 NPs
implemented in the degassed solutions. They produced and interfered with the TMBO2Co3O4 NPs system. This
dierent colors, as in the photograph shown in Figure 3b, supported the above result that the oxygen absorbed on the
and the degassed Co3O4 NPs also catalyzed the oxidation of surface of the Co3O4 NPs oxidized TMB in this system.
TMB to produce the typical color reaction. It is suggested that The oxidation of TMB is an electron transfer process.41 The
the oxygen on the surface of Co3O4 NPs was due not only to electrocatalytic behavior of a Co3O4 NP modied glassy carbon
physical adsorption. Chemisorbed oxygen probably existed on electrode (GCE) toward the electrochemical oxidation of TMB
the surface of Co3O4 NPs because of the low oxygen CoO was studied using cyclic voltammetry (CV), in order to
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Figure 6. A pHA curve (a), a temperatureA curve (b), a TMB concentrationA curve (c), and a Co3O4 NPs concentrationA curve (d)
for 6 M sulte detection, where A = A(blank,652nm) A(sulfite,652nm), in 0.2 M acetic acidacetate buer solutions incubated for 10 min with the as-
prepared Co3O4 NPs as oxidase mimics.

Figure 8. A response of TMB solutions to blank and dierent

common ions in the presence of sulte (12 M). The other ions added
Figure 7. Doseresponse curve for dierent concentrations of sulte are 240 M. Relative A: (Atotal Asulfite)/Asulfite.
standard solutions under the optimum conditions. Inset: Linear
calibration plot for sulte detection. Table 1. Results for the Determination of the Sulte in
Three Kinds of Foods
investigate the mechanism of oxidase-like catalytic activity of
Co3O4 NPs. The CVs of TMB at a bare GCE and the Co3O4 initial added average RSD
NPs-modied GCE are shown in Figure 4. Two pairs of redox amount (g recovery (n = 3)
sample (mg g1) mL1) recovery (%) (%) (%)
peaks of TMB were found under the bare GCE, and increased
current was obviously observed under the Co3O4 NPs-modied dried lily 1.7 0.5 96.399.4 98.9 0.6
bulbs 1.0 94.499.5 96.5 2.8
GCE. It is suggested that Co3O4 NPs exhibited an electro-
catalytic activity to oxidate TMB, which indicated that the 1.5 98.4100.6 100.3 2.6
Co3O4 NPs had the ability to accelerate the electron transfer.42 vermicelli 0.7 0.5 90.595.0 93.1 2.5
In order to further demonstrate that the oxidase-like activity 1.0 91.696.2 93.8 2.5
of the Co3O4 NPs was not from O2 and OH, an EPR spin 1.5 96.897.4 97.1 1.5
trapping experiment was carried out under the optimized haw roll 2.9 0.5 95.299.4 96.3 1.1
experimental conditions. The results revealed that no DMPO- 1.0 99.1101.6 100.5 3.3
O2 or DMPO-OH spin adducts were detected (Figure S7). 1.5 97.899.9 98.9 1.1

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It is suggested that TMB was not oxidized by O2 and OH graphs for the determination of sulte were constructed under
radicals in the TMBO2Co3O4 NPs system, which was the optimum conditions described above. The calibration data
dierent from the behavior of CoFe2O4 NPs.8 Therefore, the of A versus sulte concentration are shown in Figure 7. The
nature of oxidase-like activity of the Co3O4 NPs does not results indicated that the concentration of detectible sulte
originate from the O2 and OH radicals generated. could be 0.2 106 M with a linear range from 0.2 106 to
Taken together, although the detailed mechanism remains 1.6 105 M. The limit of detection (LOD) for sulte was 5.3
unclear, these observations demonstrated that Co3O4 NPs 108 M. (LOD = KS0/S, where K is a numerical factor chosen
accelerated the electron transfer from the peroxidase substrate according to the condence level desired, S0 is the standard
to oxygen absorbed on the surface of the Co3O4 NPs. The deviation (SD) of the blank measurements (n = 10, K = 3), and
possible mechanism of oxidase-like activity of Co3O4 NPs is S is the slope of the calibration curve.44)
presented in Scheme 1; that is, TMB was absorbed on the Eect of Foreign Substances. The eect of foreign
surface of the Co3O4 NPs and donated electrons to the Co3O4 substances was tested by analyzing the mixed solution and
NPs, resulting in an increase in electron density and mobility in standard solution of sulte (12 M). The molar ratio of other
the Co3O4 NPs, thus accelerating the electron transfer to ion and sulte was 20 in the mixed solution. The relative A
oxygen absorbed on the surface of the Co3O4 NPs. After the values ((Atotal Asulfite)/Asulfite) are presented in Figure 8.
completion of the electron transfer, the Co3O4 NPs reverted to As can be seen, Fe and Cu were the main interferences for
their original state. sulte determination, since trace amounts of heavy metal ions
Steady-State Kinetic Assay of Co3O4 NPs. Since Co3O4 in the water are believed to act as the catalyst in the oxidation
NPs exhibited oxidase-like activity, the kinetic parameters of of sulte by oxygen.45 EDTA was selected as a chelate reagent
only TMB/ABTS were determined by changing the concen- for the present study to eliminate the interferences derived
tration of TMB/ABTS. A typical MichaelisMenten curve from Fe, Cu, and other coexisting transient metals in the
(Figure 5a,b) was obtained with TMB/ABTS by monitoring samples.
the absorbance change at 652/417 nm for 4 min (Figure Detection of Sulte in Real Samples. As an illustration
S8a,b). The MichaelisMenten constant (Km) and maximum of analytical application, the proposed method was used to
initial velocity (Vmax) were obtained using a LineweaverBurk determine sulte in real samples under the optimal
plot. The values of Km and Vmax for the Co3O4 NPs with TMB experimental conditions. The results are listed in Table 1.
were 0.051 mM and 3.30 108 M s1. The values of Km and The recoveries of the three samples ranged from 93.8% to
Vmax for the Co3O4 NPs with ABTS were 0.037 mM and 3.20 100.5%. Hence, the proposed method can be considered a
108 M s1. The Km value of Co3O4 NPs with ABTS was useful tool for the quantication of sultes in real samples.
smaller than that with TMB. It is suggested that Co3O4 NPs In summary, Co3O4 NPs were rst found to possess oxidase-
have higher anity toward ABTS than TMB, because ABTS like activity and could catalyze the oxidation of TMB and ABTS
has a higher anity toward a positively charged nanoparticle to produce the typical color reaction in the absence of
surface and TMB has a stronger anity toward a negatively exogenously added H2O2. It was found that the oxygen
charged nanoparticle surface.43 absorbed on the surface of the Co3O4 NPs oxidizes the
Optimization of Experimental Conditions. The reaction peroxidase substrate, rather than the oxygen dissolved in the
conditions were optimized to establish the optimum analytical solution. The oxidase-like activity of Co3O4 NPs was not from
conditions. The pH value not only decides the charge of the the generated O2 or OH radical, probably originating from
surface of Co3O4 NPs (Supporting Information), which aects their ability to transfer an electron between the peroxidase
the interaction between Co3O4 NPs and sulte, but also aects substrate and oxygen absorbed on the surface of the Co3O4
the catalytic activity of Co3O4 NPs. The catalytic activity of NPs. Moreover, as the existence of sulte inhibited TMBO2
Co3O4 NPs is also dependent on the incubation temperature. Co3O4 NPs system and caused a change in color of the reaction
In addition, the concentrations of TMB and Co3O4 NPs system, a new colorimetric method for detecting sulte was
determine the color level of the reaction system. Therefore, developed. The colorimetric method showed a good response
they were investigated in this study as inuencing factors. toward sulte detection with a linear range from 0.2 106 to
Figure 6a indicates that A, where A = A(blank,652nm) 1.6 105 M and good recoveries ranging from 93.8% to
A(sulfite,652nm), increased and then decreased with increasing pH 100.5% in three real samples using Co3O4 NPs as an oxidase
and had a maximum at 4.0. The reaction temperature- mimic. The detection platform showed great potential
dependent response curve is shown in Figure 6b. It showed applications in reducing substrates. In addition, Co3O4 NPs
that A of the reaction system had a peak value at 40 C. can be expected to provide potential applications for chemical
Therefore, 4.0 and 40 C were taken as the optimal pH and sensing, biochemical analysis, electrochemical biosensing, and
temperature. The concentrations of TMB and Co3O4 NPs even clinical chemistry.

response curves are shown in Figure 6c and d; the optimal
concentrations of TMB and Co3O4 NPs were 0.25 mM and 40
g mL1, respectively.
Analytical Performance for Sulte Determination. As * Supporting Information
an oxidase mimic, Co3O4 NPs can catalyze TMB to form This material is available free of charge via the Internet at
colored product in the absence of H2O2. The sulte inhibits the

TMBO2Co3O4 NPs system and causes a change in color of
the reaction system, which suggested that sulte can be AUTHOR INFORMATION
indirectly detected. Since sulte is easily adsorbed on the
surface of Co3O4 NPs and consumes absorbed oxygen more Corresponding Author
easily than dissolved oxygen, the sulte that is consumed by *E-mail: (X.-G. Chen). Tel: 86-931-
dissolved oxygen is neglected in the reaction. The calibration 8912763 Fax: 86-931-8912582.
F | J. Agric. Food Chem. XXXX, XXX, XXXXXX
Journal of Agricultural and Food Chemistry Article

Funding peroxidase-like behavior, and electrochemical sensing of H2O2. ACS

The authors are grateful for nancial support from the National Appl. Mater. Interfaces 2012, 4, 19191927.
Natural Science Foundation of China (No. 21375053) and the (20) Xie, J.; Zhang, X.; Wang, H.; Zheng, H.; Huang, Y. Analytical
Specialized Research Fund for the Doctoral Program of Higher and environmental applications of nanoparticles as enzyme mimetics.
Education (No. 20130211110039). TrAC Trends Anal. Chem. 2012, 39, 114129.
(21) Novoselov, K. S.; Geim, A. K.; Morozov, S.; Jiang, D.; Zhang, Y.;
Notes Dubonos, S.; Grigorieva, I.; Firsov, A. Electric field effect in atomically
The authors declare no competing nancial interest.

thin carbon films. Science 2004, 306, 666669.

(22) Chen, H.; Li, Y.; Zhang, F.; Zhang, G.; Fan, X. Graphene
REFERENCES supported Au-Pd bimetallic nanoparticles with core-shell structures
(1) Bush, R. K. Sulfites in foods: uses, analytical methods, residues, and superior peroxidase-like activities. J. Mater. Chem. 2011, 21,
fate, exposure assessment, metabolism, toxicity, and hypersensitivity. 1765817661.
Adv. Food Res. 1986, 30, 176. (23) Kim, M. I.; Shim, J.; Li, T.; Lee, J.; Park, H. G. Fabrication of
(2) Sapers, G. Browning of foods: control by sulfites, antioxidants, nanoporous nanocomposites entrapping Fe3O4 magnetic nanoparticles
and other means. Food Technol. (Chicago, IL, U.S.) 1993, 47, 7584. and oxidases for colorimetric biosensing. Chem.Eur. J. 2011, 17,
(3) Adams, J. Food additive-additive interactions involving sulphur 1070010707.
dioxide and ascorbic and nitrous acids: a review. Food Chem. 1997, 59, (24) Ye, Y.; Kong, T.; Yu, X.; Wu, Y.; Zhang, K.; Wang, X. Enhanced
401409. nonenzymatic hydrogen peroxide sensing with reduced graphene
(4) Vally, H.; Carr, A.; El-Saleh, J.; Thompson, P. Wine-induced oxide/ferroferric oxide nanocomposites. Talanta 2012, 89, 417421.
asthma: a placebo-controlled assessment of its pathogenesis. J. Allergy (25) Hao, J.; Zhang, Z.; Yang, W.; Lu, B.; Ke, X.; Zhang, B.; Tang, J.
Clin. Immunol. 1999, 103, 4146. In situ controllable growth of CoFe2O4 ferrite nanocubes on graphene
(5) Filik, H.; C etintas, G. Determination of sulfite in water and dried for colorimetric detection of hydrogen peroxide. J. Mater. Chem. A
fruit samples by dispersive liquidliquid microextraction combined 2013, 1, 43524357.
with UVvis fiber optic linear array spectrophotometry. Food Anal. (26) Jiang, Y.; Wang, W.; Li, X.; Wang, X.; Zhou, J.; Mu, X. Enzyme-
Methods 2012, 5, 13621367. mimetic catalyst-modified nanoporous SiO2cellulose hybrid compo-
(6) Du, J.; Shao, Q.; Yin, S.; Jiang, L.; Ma, J.; Chen, X. Colorimetric sites with high specific surface area for rapid H2O2 detection. ACS
chemodosimeter based on diazonium-gold-nanoparticle complexes for
Appl. Mater. Interfaces 2013, 5, 19131916.
sulfite ion detection in solution. Small 2012, 8, 34123416. (27) Lin, Y.; Li, Z.; Chen, Z.; Ren, J.; Qu, X. Mesoporous silica-
(7) Pundir, C. S.; Rawal, R. Determination of sulfite with emphasis
encapsulated gold nanoparticles as artificial enzymes for self-activated
on biosensing methods: a review. Anal. Bioanal. Chem. 2013, 405,
30493062. cascade catalysis. Biomaterials 2013, 34, 26002610.
(8) Zhang, X.; He, S.; Chen, Z.; Huang, Y. CoFe2O4 nanoparticles as (28) Lin, Y.; Zhao, A.; Tao, Y.; Ren, J.; Qu, X. Ionic liquid as an
oxidase mimic-mediated chemiluminescence of aqueous luminol for efficient modulator on artificial enzyme system: toward the realization
sulfite in white wines. J. Agric. Food Chem. 2013, 61, 840847. of high-temperature catalytic reactions. J. Am. Chem. Soc. 2013, 135,
(9) Stewart, M. E.; Anderton, C. R.; Thompson, L. B.; Maria, J.; 42074210.
Gray, S. K.; Rogers, J. A.; Nuzzo, R. G. Nanostructured plasmonic (29) Wang, J.; Han, D.; Wang, X.; Qi, B.; Zhao, M. Polyoxometalates
sensors. Chem. Rev. 2008, 108, 494521. as peroxidase mimetics and their applications in H2O2 and glucose
(10) Liu, J.; Cao, Z.; Lu, Y. Functional nucleic acid sensors. Chem. detection. Biosens. Bioelectron. 2012, 36, 1821.
Rev. 2009, 109, 19481998. (30) Wang, W.; Jiang, X.; Chen, K. CePO4: Tb, Gd hollow
(11) Song, S.; Qin, Y.; He, Y.; Huang, Q.; Fan, C.; Chen, H.-Y. nanospheres as peroxidase mimic and magneticfluorescent imaging
Functional nanoprobes for ultrasensitive detection of biomolecules. agent. Chem. Commun. 2012, 48, 68396841.
Chem. Soc. Rev. 2010, 39, 42344243. (31) Wang, W.; Jiang, X.; Chen, K. Iron phosphate microflowers as
(12) Gao, L.; Zhuang, J.; Nie, L.; Zhang, J.; Zhang, Y.; Gu, N.; Wang, peroxidase mimic and superoxide dismutase mimic for biocatalysis and
T.; Feng, J.; Yang, D.; Perrett, S. Intrinsic peroxidase-like activity of biosensing. Chem. Commun. 2012, 48, 72897291.
ferromagnetic nanoparticles. Nat. Nanotechnol. 2007, 2, 577583. (32) Song, Y.; Wang, X.; Zhao, C.; Qu, K.; Ren, J.; Qu, X. Label-free
(13) Wei, H.; Wang, E. Nanomaterials with enzyme-like character- colorimetric detection of single nucleotide polymorphism by using
istics (nanozymes): next-generation artificial enzymes. Chem. Soc. Rev. single-walled carbon nanotube intrinsic peroxidase-like activity.
2013, 42, 60606093. Chem.Eur. J. 2010, 16, 36173621.
(14) Dai, Z.; Liu, S.; Bao, J.; Ju, H. Nanostructured FeS as a mimic (33) Wei, H.; Wang, E. Fe3O4 magnetic nanoparticles as peroxidase
peroxidase for biocatalysis and biosensing. Chem.Eur. J. 2009, 15, mimetics and their applications in H2O2 and glucose detection. Anal.
43214326. Chem. 2008, 80, 22502254.
(15) He, W.; Jia, H.; Li, X.; Lei, Y.; Li, J.; Zhao, H.; Mi, L.; Zhang, L.; (34) Song, Y.; Qu, K.; Zhao, C.; Ren, J.; Qu, X. Graphene oxide:
Zheng, Z. Understanding the formation of CuS concave super- intrinsic peroxidase catalytic activity and its application to glucose
structures with peroxidase-like activity. Nanoscale 2012, 4, 35013506. detection. Adv. Mater. 2010, 22, 22062210.
(16) Maji, S. K.; Dutta, A. K.; Dutta, S.; Srivastava, D. N.; Paul, P.; (35) Su, L.; Feng, J.; Zhou, X.; Ren, C.; Li, H.; Chen, X. Colorimetric
Mondal, A.; Adhikary, B. Single-source precursor approach for the
detection of urine glucose based znfe2o4 magnetic nanoparticles. Anal.
preparation of CdS nanoparticles and their photocatalytic and intrinsic
peroxidase like activity. Appl. Catal. B: Environ. 2012, 126, 265274. Chem. 2012, 84, 57535758.
(17) Maji, S. K.; Dutta, A. K.; Srivastava, D. N.; Paul, P.; Mondal, A.; (36) Ding, N.; Yan, N.; Ren, C.; Chen, X. Colorimetric
Adhikary, B. Peroxidase-like behavior, amperometric biosensing of determination of melamine in dairy products by Fe3O4 magnetic
hydrogen peroxide and photocatalytic activity by cadmium sulfide nanoparticlesH2O2ABTS detection system. Anal. Chem. 2010, 82,
nanoparticles. J. Mol. Catal. A: Chem. 2012, 358, 19. 58975899.
(18) Dutta, A. K.; Maji, S. K.; Mondal, A.; Karmakar, B.; Biswas, P.; (37) Dong, Y.; He, K.; Yin, L.; Zhang, A. A facile route to controlled
Adhikary, B. Iron selenide thin film: peroxidase-like behavior, glucose synthesis of Co3O4 nanoparticles and their environmental catalytic
detection and amperometric sensing of hydrogen peroxide. Sens. properties. Nanotechnology 2007, 18, 435602.
Actuators, B 2012, 173, 724731. (38) Zhao, J.; Xie, Y.; Yuan, W.; Li, D.; Liu, S.; Zheng, B.; Hou, W. A
(19) Dutta, A. K.; Maji, S. K.; Srivastava, D. N.; Mondal, A.; Biswas, hierarchical CoFe LDH rope-like nanostructure: facile preparation
P.; Paul, P.; Adhikary, B. Synthesis of FeS and FeSe nanoparticles from from hexagonal lyotropic liquid crystals and intrinsic oxidase-like
a single source precursor: a study of their photocatalytic activity, catalytic activity. J. Mater. Chem. B 2013, 1, 12631269.

G | J. Agric. Food Chem. XXXX, XXX, XXXXXX

Journal of Agricultural and Food Chemistry Article

(39) Tang, X.; Li, J.; Hao, J. Synthesis and characterization of spinel
Co3O4 octahedra enclosed by the {111} facets. Mater. Res. Bull. 2008,
43, 29122918.
(40) Yan, Q.; Li, X.; Zhao, Q.; Chen, G. Shape-controlled fabrication
of the porous Co3O4 nanoflower clusters for efficient catalytic
oxidation of gaseous toluene. J. Hazard. Mater. 2012, 209, 385391.
(41) Josephy, P. D.; Mason, R. P.; Eling, T. Cooxidation of the
clinical reagent 3,5,3,5-tetramethylbenzidine by prostaglandin
synthase. Cancer Res. 1982, 42, 25672570.
(42) Mu, J.; Wang, Y.; Zhao, M.; Zhang, L. Intrinsic peroxidase-like
activity and catalase-like activity of Co3O4 nanoparticles. Chem.
Commun. 2012, 48, 25402542.
(43) Yu, F.; Huang, Y.; Cole, A. J.; Yang, V. C. The artificial
peroxidase activity of magnetic iron oxide nanoparticles and its
application to glucose detection. Biomaterials 2009, 30, 47164722.
(44) Chen, J.; Chen, H.; Zhou, C.; Xu, J.; Yuan, F.; Wang, L. An
efficient upconversion luminescence energy transfer system for
determination of trace amounts of nitrite based on NaYF4Yb 3+ Er
as donor. Anal. Chim. Acta 2012, 713, 111114.
(45) Humphrey, R. E.; Ward, M. H.; Hinze, W. Spectrophotometric
determination of sulfite with 4, 4-dithio-dipyridine and 5,5-dithiobis
(2-nitrobenzoic acid). Anal. Chem. 1970, 42, 698702.

H | J. Agric. Food Chem. XXXX, XXX, XXXXXX