Professional Documents
Culture Documents
Materials Chemistry B
View Article Online
Published on 05 July 2013. Downloaded by University of East Anglia Library on 20/01/2015 09:28:39.
PAPER
excess of ascorbic acid (AA). The practical application of N-doped graphene supported PtNP materials is
eectively shown for the determination of HCY in both human blood serum and urine samples, by
dierential pulse voltammetry under optimized conditions. Our ndings conclude that N-doped
DOI: 10.1039/c3tb20923g
graphene supported PtNPs can be developed as a high performance and versatile nano-electrocatalyst
www.rsc.org/MaterialsB
Introduction
Materials Science and Engineering Program, The University of Texas at Austin, Austin,
TX 78712, USA. E-mail: maiyalagan@gmail.com
Electronic supplementary
10.1039/c3tb20923g
information
(ESI)
available.
See
DOI:
Published on 05 July 2013. Downloaded by University of East Anglia Library on 20/01/2015 09:28:39.
Experimental section
2.1
Paper
solution. 5 min later, 20 mL of 30% H2O2 was dropped into the
reaction system. Finally, the product was washed with HCl
solution and then washed three times with water. The resulting
solid was dispersed in water by ultrasonication to make
an aqueous dispersion of GNs with a concentration of about
4 mg mL1.
2.2 Preparation of the nitrogen doped graphene nanosheets
(N-GNs)
Graphene nanosheets with a high nitrogen content were
synthesized through a one-pot process using urea as the
chemical dopant.38 Typically, 10 mL graphene nanosheet
(50 mg) aqueous dispersion was diluted with 25 mL of
deionized water, and then 2 g of urea was added to the graphene
nanosheet dispersion under sonication for 3 h. Aer that, the
solution was sealed in a 50 mL Teon-lined autoclave and
maintained at 160 C for 6 h. The solids (N-doped graphene
nanosheets) were ltered and washed with distilled water
several times. Finally, the collected sample was dryed in a
vacuum oven at 60 C to give nitrogen doped graphene. The
above procedure is simpler and more versatile than preparing
nitrogen doped graphene using a plasma treatment process.39
2.3 Preparation of nitrogen doped graphene nanosheet
supported PtNPs (N-GNPtNPs)
Several reports in the literature discuss the deposition of
PtNPs.4042 In this work, the functionalization of PtNPs on the NGNs was carried out by chemical co-reduction of Pt precursor
salts along with GNs in ethylene glycol (EG)water solutions.43,44
In brief, 100 mg nitrogen doped GNs was added to 100 mL
aqueous solution containing 0.02 mM H2PtCl6, and then the
mixture was ultrasonically treated for 1.5 h to form a stable
colloid. Sequentially, 40 mL of EG was injected into the mixture
with magnetic stirring for 1 h and then the mixture was kept at
120 C for 6 h under magnetic stirring. The nal N-GNPtNP
composite was collected by ltration, washed with deionized
water, and dried in a vacuum desiccator. Our synthetic
approach is simpler than the approach recently reported by
Xiong et al.45 Graphene nanosheet supported PtNPs (GNPtNPs)
were prepared using the same procedure for comparison
purposes.
2.4
Characterization methods
Published on 05 July 2013. Downloaded by University of East Anglia Library on 20/01/2015 09:28:39.
Paper
compartment three electrode cell with a mirror polished 3 mm
glassy carbon (GC) electrode as the working electrode, Pt wire as
the counter electrode and a NaCl saturated Ag/AgCl as the
reference electrode. The electrochemical measurements were
carried out with a CHI Model 660C (Austin, TX, USA) electrochemical workstation. In cyclic voltammetry, the electrochemical oxidation of AA and HCY were carried out at a scan
rate of 50 mV s1. A pulse width of 0.05 s, an amplitude of
0.05 V, a sample period of 0.02 s and a pulse period of 0.20 s
were used in dierential pulse voltammetry (DPV). For chronoamperometric measurements, a sample interval of 0.1 s and a
potential step of 0.60 mV were used. All the electrochemical
measurements were carried out under a nitrogen atmosphere at
room temperature (27 C).
Fig. 1 FE-SEM images of GNPtNPs (A), and nitrogen doped GNPtNPs (C). The
corresponding high resolution TEM images are shown in (B) and (D) respectively.
A uniform dispersion of PtNPs on the GN support is clearly visible. The arrow
marks indicate the morphology of the GN-PtNP nanocomposite before and after
nitrogen doping. The scale bars are 100 nm (A and C) and 50 nm (B and D).
Published on 05 July 2013. Downloaded by University of East Anglia Library on 20/01/2015 09:28:39.
Fig. 2
Paper
signicantly dierent from the bulk material and bigger
PtNPs,52 and such size-dependent alteration of electronic
structures likely leads to the unusual electrocatalytic properties.
Fig. 2b depicts the core level XPS spectra of C1s, which shows a
main peak at 284.4 eV corresponding to sp2 C1s. The small
peaks observed at 285.5 eV and 287.1 eV correspond to the
formation of N-sp2 C and N-sp3 C bonds, respectively and this
would originate from the substitution of N atoms, defects or the
edge of the graphene sheets.5659 It has been reported that in
pristine graphene, the N1s peak is absent, while in the N-doped
graphene, the N1s peak was observed in three components
(Fig. 2c), indicating that the N atoms were in three dierent
bonding characters inserted into the graphene nanosheets. The
two peaks observed at 398.37 and 400.76 eV correspond to the
pyridinic and pyrrolic N atoms, respectively. These refer to
the N atoms which were located in a p conjugated system and
contribute to the p system with one or two p-electrons,
respectively.56,60,61 Moreover, it has been reported that for the
pyridinic-N, the nitrogen atom contributes one p-electron to the
aromatic p-system and has a lone electron pair in the plane of
the ring. As for the pyrrolic-N, the nitrogen atom contributes
two p-electrons to the p-system, and a hydrogen atom is bound
in the plane of the ring, to support the graphene skeleton
nanosheets.56,62 Furthermore, a peak at 401.58 eV corresponds
to graphitic N, which refers to the N atoms that replaced the C
Core level XPS spectra of (a) Pt (4f), (b) C1s, and (c) N1s orbitals of nitrogen doped GNPtNP nanocomposites.
Published on 05 July 2013. Downloaded by University of East Anglia Library on 20/01/2015 09:28:39.
Paper
(1)
n t/d002
(2)
Fig. 3 XRD patterns of GNPtNPs (a) and N-GNPtNP nanocomposites (b). Inset
is the XRD pattern of the graphene nanosheets.
(3)
Published on 05 July 2013. Downloaded by University of East Anglia Library on 20/01/2015 09:28:39.
Paper
near-physiological conditions (pH 7.2), HCY has an amino
group NH3+ and a carboxyl group CO2.73,74 At this point, the
electrostatic interaction between the negatively charged CO2
group of HCY and the positively charged pyridinium moieties of
the N-doped GNPtNPs gave a higher oxidation current for HCY.
It has also been implied that the incorporated nitrogen atoms
can enhance the interaction between the carbon structure and
metals, thus the kinetics of HCY diusion and biosensing eciency can be improved. Moreover, PtNPs have a large specic
active surface area and can act as tiny conducting centers, which
were distributed throughout the GN network and formed a
continuous assembly of PtNPs on the surface, resulting in a
decrease in the energy barrier, facilitating electron transfer, and
improving biosensor response. It has been shown that the thiol
(SH) and amino group (NH3+) in the HCY molecule have a
higher anity for PtNPs in the nanocomposite, through PtS
and interparticle binding and electrostatic interactions between
amino groups. This means that more HCY biomolecules can
access the PtNP surface by three-dimensional assembly
(Scheme 2), as compared to the two-dimensional substrate. To
understand the fast electron transfer reaction of HCY at the NGNPtNP modied electrode quantitatively, we have calculated
the standard heterogeneous rate constant (ks) for HCY at N-GN
PtNP modied electrodes and other electrodes. HCY oxidation
is an irreversible process and hence we have used Velasco eqn
(4) (ref. 75) to calculate the heterogeneous rate constant (ks):
ks 1.11Do1/2(Ep Ep1/2)1/2y1/2
(4)
Fig. 4 (A) CVs obtained for 0.2 mM HCY at (a) a bare GC electrode and (b) GN, (c) GNPtNP and (d) N-GNPtNP modied GC electrodes in 0.2 M PB solution (pH 7.2
N-GNs) at a scan rate of 50 mV s1. (e) CV of a N-GNPtNP modied GC electrode in the absence of 0.2 mM HCY. (f) CV of a N-GN modied GC electrode toward the
oxidation of HCY. (B) Linear sweep voltammograms (LSVs) obtained for 0.2 mM HCY at a N-GNPtNP modied electrode in 0.2 M PB solution at dierent scan rates: (a)
25, (b) 50, (c) 75, (d) 100, (e) 125, (f) 150, (g) 175, (h) 200, (i) 225 and (j) 250 mV s1. The inset calibration plot is obtained by plotting the square root of the scan rate vs.
the anodic oxidation current of HCY.
Published on 05 July 2013. Downloaded by University of East Anglia Library on 20/01/2015 09:28:39.
Paper
Fig. 5 CVs obtained for a mixture of 0.2 mM each of HCY and AA at (a) a bare GC
electrode and (b) GN, (c) GNPtNP and (d) N-GNPtNP modied GC electrodes in
0.2 M PB solution (pH 7.2) at a scan rate of 50 mV s1. Curve (e) corresponds to
the CV of a N-GNPtNP modied GC electrode after the 20th measurement and
(f) is the CV of a N-GNPtNP modied GC electrode in the absence of HCY and AA.
Published on 05 July 2013. Downloaded by University of East Anglia Library on 20/01/2015 09:28:39.
amperometric current response was observed due to the addition of 2 nM HCY. Then with the addition of a further 2 nM HCY
in each step, with a sample interval of 50 s, the current response
linearly increased and a steady state current response was
obtained within 2 s, which indicates a fast electron-transfer
process at this electrode. We found that a 2.40 nA current was
obtained for the single addition of 2 nM HCY (1.35 nA nM1) at
the N-GNPtNP electrode. The observed stable amperometric
current response together with higher sensitivity to HCY of the
N-GNPtNP modied electrode indicates that this electrode can
be successfully used for the sensitive detection of 2 nM HCY.
The amperometric current responses increased linearly with
HCY concentration from 2 to 24 nM (Fig. 7A; curve a) and the
calibration plot obtained for amperometric current responses
vs. various concentrations of HCY is shown in the inset of
Fig. 7A. A good linearity was obtained with a correlation coefcient of 0.9970 in the concentration range that was used in
Paper
Fig. 7. For comparison, the sensitivity of the GNPtNP modied
electrode towards HCY was examined by amperometry and the
sensitivity limit was found to be 0.6 nA nM1 (Fig. 7 curve b).
Moreover, we have observed amperometric signals with picomolar concentrations of HCY, as shown in Fig. 7B. The N-GN
PtNP electrode was polarized at +0.60 V, and aliquots of HCY
were injected into the stirred electrolyte solution. A rapid
increase in the current was noticed aer each addition of 200
pM HCY into the electrolyte solution, and a steady state
response was attained within 2 s. Thus the experimental
detection limit of the present N-GNPtNP modied electrode
was calculated to be 200 pM. The N-GNPtNP electrode is highly
sensitive, and the amperometric response is very stable and
oers a linear dependence over a wide range of HCY concentrations (1 mM). The observed results indicate that the N-GN
PtNP modied electrode showed higher sensitivity than the GN
PtNP modied electrode. It is worth comparing the performance of the N-GNPtNP modied electrode towards HCY with
those of recently reported polymer, polymer-nanoparticles and
other chemically modied electrodes. We summarized the
limits of detection with relevant references in ESI, Table S1.
Our limit of detection is very low (200 pM), compared to the
reports available in the literature based on other electrochemical HCY biosensors. In our system, PtNPs were embedded
three dimensionally onto the surface of the nitrogen doped
GNs, leading to the formation of more electroactive graphene
layers and electrostatic interactions between HCY and the three
dimensionally formed N-GNPtNP, which oers a high performance platform toward the detection of HCY.
3.7
Eect of interferents
Fig. 7 (A) Amperometric it curves for each addition of 2 nM HCY at N-GNPtNP (a) and GNPtNP (b) modied GC electrodes in 0.2 M PB solution (pH 7.2) at a
regular time interval of 50 s. (B) The N-GNPtNP electrode with 200 pM of HCY added into the stirred PB solution at regular time interval of 50 s. The electrode was
polarized at +0.60 V. The inset shows the corresponding calibration plot.
Published on 05 July 2013. Downloaded by University of East Anglia Library on 20/01/2015 09:28:39.
Paper
response was due to the addition of 2 nM HCY (a) and with the
further addition of 2 nM HCY in each step with a sample
interval of 100 s, the current response increases and a steady
state current response was attained within 2 s (a and b). Aer
two steps (a and b),1 mM each of glucose (c), urea (d), uric acid
(e) serotonin (f), oxalate (g) and thiourea (h) were added separately with a sample interval of 50 s to the same 0.2 M PB
solution (pH 7.2), no change in amperometric current
response was observed. However, with the addition of 2 nM
HCY to the same solution (i, j and k), the current response was
again increased similar to steps a and b. Similarly, the selectivity of HCY was also studied in the presence of some other
important biomolecules (interferents) such as dopamine,
epinephrine and L-dopa (Fig. S2 in ESI); the N-GNPtNP
modied electrode was highly selective toward the determination of HCY in the presence of important biomolecules. The
observed results indicate that the N-GNPtNP modied electrode can be successfully used for the determination of 2 nM
HCY, even in the presence of a 500-fold excess of several
common interferents.
3.8
Fig. 9 Plot of current vs. time for the oxidation of HCY (0.2 mM) at the N-GN
PtNP electrode in 0.2 M PB solution, demonstrating the operational stability of the
electrode.
Published on 05 July 2013. Downloaded by University of East Anglia Library on 20/01/2015 09:28:39.
Paper
Fig. 10 (A) DPVs obtained (a) for human blood serum and (b) after the addition of 20 mM commercial HCY to human blood serum, for the N-GNPtNP modied
electrode in 0.2 M PB solution. (B) DPVs obtained (a) for human urine and (b) after the addition of 20 mM commercial HCY to human urine, for the N-GNPtNP modied
electrode in 0.2 M PB solution from 0 to 0.70 V. Pulse width 0.05 s, amplitude 0.05 V, sample period 0.02 s and pulse period 0.2 s.
Conclusions
blood serum and urine samples by dierential pulse voltammetry under optimized conditions. Our ndings conclude that
N-GNPtNP nanocomposites can be developed as ecient and
versatile high performance electrocatalysts for electrochemical
biosensor applications.
Acknowledgements
Palanisamy Kannan and Marcin Opallo thank NanOtechnology
Biomaterials and aLternative Energy Source for ERA Integration
[FP7-REGPOT-CT-2011-285949-NOBLESSE] Project from European Union.
References
1 A. J. Patil, J. L. Vickery, T. B. Scott and S. Mann, Adv. Mater.,
2009, 21, 31593164.
2 C.-H. Lu, H.-H. Yang, C.-L. Zhu, X. Chen and G.-N. Chen,
Angew. Chem., Int. Ed., 2009, 48, 47854787.
3 N. Mohanty and V. Berry, Nano Lett., 2008, 8, 44694476.
4 M. Zhou, Y. Zhai and S. Dong, Anal. Chem., 2009, 81, 5603
5613.
5 Y. Wang, J. Lu, L. Tang, H. Chang and J. Li, Anal. Chem.,
2009, 81, 97109715.
6 C. Shan, H. Yang, J. Song, D. Han, A. Ivaska and L. Niu, Anal.
Chem., 2009, 81, 23782382.
7 Y. Shao, J. Wang, H. Wu, J. Liu, I. A. Aksay and Y. Lin,
Electroanalysis, 2010, 22, 10271036.
8 X. Li, G. Zhang, X. Bai, X. Sun, X. Wang, E. Wang and H. Dai,
Nat. Nanotechnol., 2008, 3, 538542.
9 A. K. Geim and K. S. Novoselov, Nat. Mater., 2007, 6, 183191.
10 H. Huang, S. Chen, X. Gao, W. Chen and A. T. S. Wee, ACS
Nano, 2009, 3, 34313436.
11 T. Ramanathan, A. A. Abdala, S. Stankovich, D. A. Dikin,
M. Herrera Alonso, R. D. Piner, D. H. Adamson,
H. C. Schniepp, X. Chen, R. S. Ruo, S. T. Nguyen,
I. A. Aksay, R. K. Prud'Homme and L. C. Brinson, Nat.
Nanotechnol., 2008, 3, 327331.
Published on 05 July 2013. Downloaded by University of East Anglia Library on 20/01/2015 09:28:39.
Paper
12 J. L. Vickery, A. J. Patil and S. Mann, Adv. Mater., 2009, 21,
21802184.
13 H. Wang, T. Maiyalagan and X. Wang, ACS Catal., 2012, 2,
781794.
14 H. M. Jeong, J. W. Lee, W. H. Shin, Y. J. Choi, H. J. Shin,
J. K. Kang and J. W. Choi, Nano Lett., 2011, 11, 24722477.
15 Y. Ma, A. S. Foster, A. V. Krasheninnikov and
R. M. Nieminen, Phys. Rev. B: Condens. Matter Mater. Phys.,
2005, 72, 205416.
16 C. Zhou, J. Kong, E. Yenilmez and H. Dai, Science, 2000, 290,
15521555.
17 T. O. Wehling, K. S. Novoselov, S. V. Morozov, E. E. Vdovin,
M. I. Katsnelson, A. K. Geim and A. I. Lichtenstein, Nano
Lett., 2007, 8, 173177.
18 L. Liu, S. Ryu, M. R. Tomasik, E. Stolyarova, N. Jung,
M. S. Hybertsen, M. L. Steigerwald, L. E. Brus and
G. W. Flynn, Nano Lett., 2008, 8, 19651970.
19 G. Giovannetti, P. A. Khomyakov, G. Brocks, V. M. Karpan,
J. van den Brink and P. J. Kelly, Phys. Rev. Lett., 2008, 101,
026803.
20 S. U. Lee, R. V. Belosludov, H. Mizuseki and Y. Kawazoe,
Small, 2009, 5, 17691775.
21 L. L. Zhang, X. Zhao, H. Ji, M. D. Stoller, L. Lai, S. Murali,
S. McDonnell, B. Cleveger, R. M. Wallace and R. S. Ruo,
Energy Environ. Sci., 2012, 5, 96189625.
22 F. Su, C. K. Poh, J. S. Chen, G. Xu, D. Wang, Q. Li, J. Lin and
X. W. Lou, Energy Environ. Sci., 2011, 4, 717724.
23 K. Xiao, Y. Liu, P. a. Hu, G. Yu, Y. Sun and D. Zhu, J. Am.
Chem. Soc., 2005, 127, 86148617.
24 E. P. Randviir and C. E. Banks, RSC Adv., 2012, 2, 58005805.
25 D. A. C. Brownson, C. W. Foster and C. E. Banks, Analyst,
2012, 137, 18151823.
26 X. Wang, X. Li, L. Zhang, Y. Yoon, P. K. Weber, H. Wang,
J. Guo and H. Dai, Science, 2009, 324, 768771.
27 P. Wu, Z. Cai, Y. Gao, H. Zhang and C. Cai, Chem. Commun.,
2011, 47, 1132711329.
28 D. Usachov, O. Vilkov, A. Gr
uneis, D. Haberer, A. Fedorov,
V. K. Adamchuk, A. B. Preobrajenski, P. Dudin, A. Barinov,
M. Oehzelt, C. Laubschat and D. V. Vyalikh, Nano Lett.,
2011, 11, 54015407.
29 L. Qu, Y. Liu, J.-B. Baek and L. Dai, ACS Nano, 2010, 4, 1321
1326.
30 A. L. M. Reddy, A. Srivastava, S. R. Gowda, H. Gullapalli,
M. Dubey and P. M. Ajayan, ACS Nano, 2010, 4, 6337
6342.
31 Z.-J. Lu, S.-J. Bao, Y.-T. Gou, C.-J. Cai, C.-C. Ji, M.-W. Xu,
J. Song and R. Wang, RSC Adv., 2013, 3, 39903995.
32 H. Yin, H. Tang, D. Wang, Y. Gao and Z. Tang, ACS Nano,
2012, 6, 82888297.
33 J. Lu, I. Do, L. T. Drzal, R. M. Worden and I. Lee, ACS Nano,
2008, 2, 18251832.
34 S. Guo, D. Wen, Y. Zhai, S. Dong and E. Wang, ACS Nano,
2010, 4, 39593968.
35 V. B. Parambhath, R. Nagar and S. Ramaprabhu, Langmuir,
2012, 28, 78267833.
36 B. P. Vinayan, K. Sethupathi and S. Ramaprabhu, Int. J.
Hydrogen Energy, 2013, 38, 22402250.
Published on 05 July 2013. Downloaded by University of East Anglia Library on 20/01/2015 09:28:39.
Paper
69 W. L. D. M. Nelan, H. J. Blom, E. A. P. Steegers, M. Den
Heijer, C. M. G. Thomas and T. K. A. B. Eskes, Obstet.
Gynecol., 2000, 95, 519524.
70 S. Biasioli and R. Schiavon, Blood Purif., 2000, 18, 177182.
71 M. M. Rees and G. M. Rodgers, Thromb. Res., 1993, 71, 337
359.
72 G. Yang, Y. Li, R. K. Rana and J.-J. Zhu, J. Mater. Chem. A,
2013, 1, 17541762.
73 I. I. S. Lim, W. Ip, E. Crew, P. N. Njoki, D. Mott, C.-J. Zhong,
Y. Pan and S. Zhou, Langmuir, 2007, 23, 826833.
74 J.-M. Zen, A. S. Kumar and J.-C. Chen, Anal. Chem., 2001, 73,
11691175.
75 J. G. Velasco, Electroanalysis, 1997, 9, 880882.
76 A. J. Bard and R. W. Faulkner, Electrochemical Methods,
Fundamentals and Applications, Wiley, New York, 2nd edn,
2001.