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Graphenesilver nanohybrids for ultrasensitive

surface enhanced Raman spectroscopy: size
Cite this: J. Mater. Chem. C, 2014, 2,
dependence of silver nanoparticles
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6850

Yazhou Zhou,ab Xiaonong Cheng,a Dan Du,b Juan Yang,*a Nan Zhao,a
Shuangbiao Ma,a Tao Zhonga and Yuehe Lin*b

Received 31st March 2014
Accepted 16th June 2014
DOI: 10.1039/c4tc00658e
www.rsc.org/MaterialsC
The surface-enhanced Raman scattering (SERS) mechanism of graphenesilver (GEAg) nanohybrids was
investigated by studying the Raman signals of rhodamine 6G (R6G) adsorbed onto nanohybrids with
dierent sizes of Ag nanoparticles (NPs). The results showed that the SERS intensity ratios between GE
Ag NPs with Ag NPs with sizes of 9, 25 and 35 nm and pure Ag substrate were estimated to be 2.7, 5.0
and 8.9 respectively. GE sheets did improve the SERS activity due to their high adsorption ability, strong
uorescence quenching and additional chemical enhancement mechanism (CM)-based SERS eect.
However, Ag NPs with relatively larger size (
nm) play a major role in SERS activity of molecules
adsorbed on the nanohybrids. Furthermore, the free-standing GEAg NP nanohybrid lm was fabricated
through the vacuum ltration of the optimized GEAg NP composite suspension followed by thermal
annealing. The as-prepared lm has the larger size of Ag NPs and narrower interparticle distance, which
provides more hot spots for sensitive detection of R6G molecules, with a detection limit at 1 
1012 M. Additionally, the relative standard deviation (RSD) values are measured to be below 0.13,
revealing a good consistency and reproducibility across the entire area of the lm, suggesting this
substrate holds a promise for routine SERS detection.

decreased from monolayer to multilayers.13,14 Compared to the

1. Introduction
Graphene (GE) has attracted increasing attention from the
physical, chemical, and biomedical elds as a novel nano-
material with many exceptional features including unparalleled
thermal conductivity, excellent electrical conductivity, high
surface-to-volume ratio, remarkable mechanical strength, and
biocompatibility.14 One of the important themes in recent
GE-based research is to investigate its surface-enhanced Raman
scattering (SERS) properties.57 In previous reports, due to its
strong optical transmission, surface plasmon range in ter-
ahertz, strong adsorption ability, large specic surface area and
ecient uorescence quenching, GE has been shown to have a
clear Raman enhancing eect for probe molecules.810 Although
SERS eects of GE were based on the deposition of molecules on
GE sheets, the enhanced eciency is also closely related to the
number of layers of GE by a chemical enhancement mechanism
(CM).11,12 This so-called n-layer GE lm gave the clear CM-based
SERS eect for molecules and the SERS intensity has drastically
a
School of Materials Science and Engineering, Jiangsu University, Zhenjiang, 212013,
Jiangsu, PR China. E-mail: yangjuan6347@mail.ujs.edu.cn
b
School of Mechanical and Materials Engineering, Washington State University,
Pullman, WA 99164-2920, USA. E-mail: Yuehe.lin@wsu.edu
Electronic supplementary information (ESI) available.
10.1039/c4tc00658e
CM, the electromagnetic mechanism (EM) is really a long-range
eect and dependent on the enhanced electromagnetic elds
generated at the interface between the investigated molecules
and the metal substrates with appropriate morphologies.15
Therefore, gold nanoparticles (Au NPs) or silver nanoparticles
(Ag NPs) were used to improve the SERS intensity of the GE lm
by deposition onto the n-layer GE lm.14,16,17 However, fabri-
cating GE devices with a single layer is not eective, which is
unsuitable for routine SERS applications.
Due to its large specic surface area, remarkable electrical
conductivity, excellent adsorptivity and high chemical and
thermal stability, GE can be used to fabricate the GEAg NP
hybrids by an in situ reduction method or self-assembly
method, which can be used for molecule detection analysis.18,19
For example, Fu et al. reported the sandwich structure of Ag NP
GOAg piece, in which 3-mercaptopropyltrimethoxysilane sily-
lated graphene oxide (GO) played as a bridge to connect Ag NPs
with Ag piece;20 Min et al. reported that the Ag NP/RGO hybrid
lms protected by GO sheets were fabricated by electrostatic
adsorption of Ag NPs on polyallylamine hydrochloride (PAA)
functionalized RGO (PAARGO) lms and subsequent electro-
static adsorption of GO sheets on the PAA functionalized Ag NP/
PAARGO lms;21 and Zheng et al. reported that the b-lacto-
See DOI:
globulinreduced GO/Au hybrid can be used as a SERS
substrate.22 By decorating Ag NPs on GE sheets, the coupled EM

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eect caused by the aggregation of the nanoparticles and strong
electronic interactions between Ag NPs and the GE sheets are
considered to be responsible for the signicantly enhanced
Raman signal of the analytes.19 However, the SERS eects of the
GEAg NP nanohybrids on molecules are very complicated. And
the SERS mechanism of these substrates is not yet fully
understood. The polymer or organic molecules used to fabricate
the hybrid SERS substrates might make the Raman enhance-
ment mechanism even more complicated. Wu et al. have
investigated the SERS eects of GE/Au or Ag hybrid aqueous
solutions, including SERS eect of noble nanoparticles and GE
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in the GOAg+ precursor were transformed to Ag NPs by the
sonication process. Aer the chemical reduction, peak C (002)
in the XRD patterns of various samples shied to around 24 ,
suggesting the reduction of GO (Fig. S1b, ESI). In addition, the
intensities of the characteristic peaks for the cubic Ag crystal
became higher than that in the GOAg+ precursor, indicating
the reduction of Ag+ to Ag. The XRD results indicated successful
preparation of GEAg NP hybrids during our experiments.
Transmission electron microscopy (TEM) experiments were
carried out to observe the morphology of GEAg NP nano-
hybrids. The typical TEM images of various samples are shown

sheets, species and concentration of the probe molecules.23 in Fig. 1ac. For each sample, it was found that the main
They pointed out that the SERS eects of both the Au or Ag NPs components in such a composite consisted of GE sheets coated
on the absorbed probe molecules and on GE, as well as that of by Ag NPs. The almost transparent GE sheets were fully exfoli-
GE on the probe molecules, were competitive. However, the ated and decorated with spherical-like Ag NPs and no obvious
SERS eect for GEAg NP nanohybrids is greatly dierent from aggregation or free NPs were detected. These GE sheets indeed
its colloid. Importantly, because of the high solvability of act as substrates for the deposition of Ag NPs, which can prevent
composites and probe molecules in water, the Raman the Ag NPs from aggregation through the so-called in situ
enhancement is relatively weak.24 It has a low eciency for the reduction method. Meanwhile, the attached particles may
detection of molecules with a low concentration (the minimum prevent the restacking of carbon sheets, and therefore the
concentration is about 104 M). More importantly, the repro- resulting composite can also be dispersed in water by sonica-
ducibility of the Ag NP decorated GE substrates in previous tion, forming a stable suspension.
reports is very poor, which limits their application in routine The average size and particle size distribution of Ag NPs were
SERS detection. We consider that the EM-based eect of Ag NPs inuenced by the reaction conditions and the average
is dominant in all SERS eects because of its large range particle size was observed in the order of GAg(SB) < GAg(Hy) <
enhancement.15 Therefore, investigation of the SERS eect of GAg(EG) (Fig. 1a*c*). For the GAg(SB) sample, sodium
GEAg NP nanohybrids by Ag NPs is useful to further clarify the
SERS mechanism and design highly sensitive and reproducible
GEAg NP nanohybrids for SERS applications.
In our previous work, we reported the design and assembly
of GEAg NP nanohybrids which have excellent optical, elec-
trical and antimicrobial properties.25,26 It is well-known that the
reduction ability of reducer has a great inuence on the
reduction degree of GO and the size of Ag NPs in GEAg NP
nanohybrids synthesized through the in situ reduction method.
In this paper, in order to study the SERS mechanism of the GE
Ag NP nanohybrids, we designed three dierent kinds of GEAg
NP nanohybrids, synthesized by using sodium borohydride,
hydrazine and ethylene glycol as reducers (labeled as GAg(SB),
GAg(Hy) and GAg(EG)). Our results show that the EM-based
eect of Ag NPs is dominant in all SERS eects, which is
dependent on the size of Ag NPs. Furthermore, we fabricated
the large-scale and exible free-standing GEAg NP nanohybrid
lms with high SERS sensitivity and good reproducibility
through vacuum ltration and followed by a thermal annealing
process.

2. Results and discussion

2.1. Synthesis of GEAg NP hybrids with dierent sizes of Ag
NPs
The formation of GEAg NP nanohybrids was characterized by
X-ray diraction (XRD). The XRD pattern of the GOAg+
precursor showed that the weak characteristic peaks centered at
38.1 , 44.3 , 64.4 and 77.4 (Fig. S1a, ESI), which can be Fig. 1 The typical TEM images of (a) GAg(SB), (b) GAg(Hy) and (c)
assigned to the (111), (200), (220) and (311) planes of the cubic GAg(EG); and (a*c*) are their particle size distributions for Ag NPs,
Ag crystal (JCPDS no. 04-0783). We concluded that a few of Ag+ correspondingly.

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borohydride with strong reduction ability was used as a reducer,
and the reducing rate of the Ag+ was very fast and the whole
reaction nished in short time at room temperature. It made
the Ag nucleate rapidly and grew to a small size (9 nm) with
narrow particle size distribution (Fig. 1a*). The GAg(Hy)
sample was fabricated through the solvothermal method and
hydrazine was used as a reducer at 140  C for 2.5 h. Similar to
GAg(SB), Ag also nucleated rapidly because of the high
reduction activity of hydrazine and grew to a small size (25 nm)
(Fig. 1b*). When the hydrazine was replaced by ethylene glycol,
the particle size of Ag NPs in the GAg(EG) sample became
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larger (35 nm) as shown in Fig. 1c*. Due to the low reducing
activity of EG, the nucleation rate of Ag NPs decreased and the
particle could grow to a big size in the sucient time. In
addition, the particle size distribution of Ag NPs in GAg(Hy)
and GAg(EG) samples was very wide according to their high
standard deviations (SDs). The SD of particle size was calculated
to be about 1.5 nm for GAg(SB), 5.7 nm for GAg(Hy) and
7.2 nm for GAg(EG) (Fig. 1a*c*).
During the chemical reduction, the structure of GO signi-
cantly changed, including the removal of oxygen from GO
sheets in the presence of a reducer. We have employed X-ray
photoelectron energy spectra (XPS) to analyze the chemical
composition and the amount of carboncarbon/carbonoxygen
bonding of GO and the GEAg NP nanohybrids, which can be
Fig. 2 The typical C1s spectra of (a) GO, (b) GAg(SB), (c) GAg(Hy)
and (d) GAg(EG) nanohybrids.
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seen in Fig. 2. Briey, the C1s XPS spectrum of GO (Fig. 2a)
clearly indicated a considerable degree of oxidation with four
components that correspond to carbon atoms in dierent
functional groups: the nonoxygenated ring C, the C in CO
bonds, the carbonyl C, and the carboxylate carbon (OC(O)).27,28
Through the chemical reaction, the oxygen functionalities in
the GEAg NP nanohybrids decreased, and their peak intensi-
ties were much lower than those in GO (Fig. 2bd). The dier-
ences of the XPS spectra among the GO and GEAg NP hybrids
in terms of elemental composition are summarized in Table S1
(Table S1, ESI). However, the structural changes have been
greatly inuenced by the dierent kinds of reducers. The GE in
GAg(SB) and GAg(Hy) hybrids has a high reducing degree
because of the strong reducing activity of NaBH4 and N2H4. The
C/O for GAg(SB) and GAg(Hy) hybrids was 4.4 and 5.9,
respectively. Additionally, there was an additional component
at 285.63 eV corresponding to C bound to nitrogen in GAg(Hy),
which is in accordance with previous reports.28 For GAg(EG)
composites, as prepared using ethylene glycol, the reducing
degree was lower than those in GAg(SB) and GAg(Hy) (the C/O
was 3.9).
The signicant structural changes occurred during the
chemical processing from the GOAg+ precursor to GEAg NP
nanohybrids, which was reected in their Raman spectra
(Fig. 3). The Raman spectrum of GO, as expected, displayed a
prominent G peak around 1600 cm1 and a D peak around
1350 cm1, corresponding to the E2g phonon of C sp2 atoms and
a breathing mode of k-point phonons of A1g symmetry (Fig. S2,
ESI).27 Aer chemical reduction by N2H4, the G and D peaks
(G(Hy) sample) shied to 1590 cm1 and 1349 cm1, corre-
spondingly. In addition, the intensity ratio of D/G (ID/IG)
increased to 1.22 compared with that of GO (0.78) (Table S2,
ESI). Meanwhile, the ID/IG for GAg(SB) and GAg(Hy) also
increased. This change suggested the decrease in the average
size of the sp2 domains, but more numerous in numbers upon
reduction of the exfoliated GO, which was consistent with
previous reports.27 However, for GAg(EG) composites, the ID/IG
increased to 0.92, lower than those in GAg(SB) and GAg(Hy).
This can be explained since the reduction activity of ethylene
The typical Raman spectra of G(Hy), GAg(SB), GAg(Hy) and
Fig. 3
GAg(EG).
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glycol is moderate, which reduced a certain degree of oxygen

functionalities. Therefore, the number of new graphitic
domains created in the GAg(EG) hybrids is smaller compared
with GAg(SB) and GAg(Hy) prepared using the strong
reducing activity of sodium borohydride and hydrazine.

2.2. SERS eects of GEAg NP nanohybrids

It is well-known that the Raman signals of GE can be enhanced
by Au NPs and Ag NPs, which mainly arise from the EM eect
based on the charge transfer between GE and the Au or Ag
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NPs.16 This can also be observed clearly in our experiment

(Fig. 3). However, an enhancing eect of the Raman signals of
GE in three GEAg NP nanohybrids varied signicantly. The
enhancement factor was estimated to be 23.4 for GAg(EG),
13.6 for GAg(Hy) and 2.2 for GAg(SB), compared with the pure
G(Hy) of the G peak intensity. In our system, the GEAg NP
nanohybrids were prepared through reduction of the same GO
Ag+ precursor and the weight percent of Ag NPs in three GEAg
NP hybrids was similar, according to the thermogravimetric
analysis (TGA) (Fig. S3, ESI). Therefore, it can be concluded
that the particle size of Ag gave a great contribution to the
enhancement activity. According to previous work,14 the
optimum size of Ag NPs for SERS activity is around 100 nm.
When the size is below 100 nm, the SERS signal increases with
an increase of the size. This is well consistent with our results:
the enhancement factor of GEAg NP nanohybrids was observed
in the order: GAg(EG) > GAg(Hy) > GAg(SB).
To further investigate the SERS activity of GEAg NP nano-
Fig. 4 (a) SERS spectra of R6G molecules with 1  106 M concen-
hybrids, rhodamine 6G (R6G) was used as the model probe. In tration adsorbed on GAg(EG) hybrids and pure G(Hy); for
our study, the laser excitation wavelength was xed at 532 nm as comparison, Raman spectrum of R6G solid is presented. (b) SERS
R6G has a very strong uorescence background compared to spectra of R6G molecules adsorbed on GAg(Hy), GAg(SB) nano-
other excitation wavelengths. Therefore, it is dicult to obtain hybrids and pure Ag NPs lm.
the typical Raman spectrum of the R6G in solution at 532 nm
excitation. Here, R6G solid was used for comparison, and the
Raman spectrum is shown in Fig. 4a. The typical characteristic (Fig. 4b). For clarity, we dened the ratio (IGEAg/IAg-pure) as the
peaks of R6G molecules were very weak. In contrast, for R6G SERS intensity ratio to investigate the Raman enhancement
adsorbed on GAg(EG), SERS signals were observed and the mechanism. The SERS intensity ratios and the assignments of
characteristic peaks can be clearly distinguished (Fig. 4a). In the Raman peaks of the R6G probe molecules are summarized
addition, some Raman signals (such as ca. 1600, 1310, 930, 662 and compared in Table 1. It can be observed that GAg(EG) has
and 638 cm1) can also be found in the SERS spectrum, while the stronger enhancement eect on R6G molecules than that of
these Raman peaks were not observed in the Raman spectrum pure Ag NPs, which indicated that the GE sheets indeed
of pure R6G solid, indicating signicant SERS activity of amplied the SERS signals of the R6G probe molecules. The
GAg(EG) hybrids. So what is the mechanism of this unique result is consistent with the precious work by Lu.30
feature? To have a better understanding of the mechanism, it is It was reported that the EM is based on surface plasmon
necessary to distinguish all the possible SERS eects in the R6G/ resonance with an enhancement factor usually in the range of
GAg(EG) composites of the probe molecules, GE sheets and Ag 106 to 108, even up to 1014, whereas the factor of CM is usually in
NPs. In this case, the pure G(Hy) and Ag NPs synthesized using the range of 10100.31 Therefore, the EM-based SERS eect of Ag
a modied procedure in previous work were used for compar- NPs on R6G molecules is still dominant in all the possible SERS
ison.25,29 The detailed characterization of the pure G(Hy) and eects. In order to further investigate the SERS eect of Ag NPs,
Ag NP colloid is provided in Fig. S4 and S5. Importantly, the the SERS spectra of R6G/GAg(SB) and R6G/GAg(Hy) were
particle size of Ag NPs in colloid was controlled to about 35 nm, measured by using the same conditions, which can be seen in
which is similar to that in GAg(EG) composites (Fig. S5, ESI). Fig. 4b. Obviously, both GAg(SB) and GAg(Hy) also have
For the R6G adsorbed on pure G(Hy) sheets, it showed a very higher SERS intensities on R6G molecules than that of pure Ag
weak SERS eect on the R6G molecules, based on the CM eect NPs, but the weaker enhancement eect than that of GAg(EG).
of GE (Fig. 4a). For pure Ag NPs, there existed an obvious SERS In addition, the IGEAg/IAg-pure can be estimated to be about
eect of the Ag NPs on the R6G molecules based on the EM 8.9 for GAg(EG), 5.0 for GAg(Hy) and 2.7 for GAg(SB),

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Table 1 SERS intensity ratio (IGEAg/IAg-pure) between the GEAg NPs

and pure Ag NPs substrates in dierent Raman peaks of R6G mole-
cules and the assignments of several Raman banda

IGEAg/IAg-pure ratio in the dierent Raman band of R6G

Aromatic CC Aromatic CC CCC ring

stretching stretching in-plane
(1646 cm1) (1363 cm1) (611 cm1)

GAg(SB) 1.8 2.7 2.2

GAg(Hy) 2.7 5.0 4.6
GAg(EG) 5.4 8.9 7.0
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a
The assignments of the peaks of R6G are based on previous reports:
ref. 3.

according to the CC stretching peak at ca. 1363 cm1 (Table 1).

We considered that the size of Ag NPs in composites was the
main contribution to SERS enhancement. The size of Ag NPs in
the GAg(EG) composites was larger than that in the other
samples, which causes the SERS intensity of GAg(EG) to be
signicantly higher by the EM-based eect of Ag NPs than that
in GAg(SB) and GAg(Hy) hybrids.
Interestingly, we found that the adsorption ability of the
three dierent kinds of GEAg NP nanohybrids for R6G mole-
cules was dierent based on SERS measurement results. We
measured the change of absorption intensity of R6G solution
(1  105 M) treated with GEAg NP nanohybrids by Ultraviolet- Fig. 5 (a) UV-vis spectra of R6G (1  105 M) and the R6G solutions
visible (UV-Vis) absorption spectra at 527 nm (Fig. 5a). The treated by adsorption with GAg(SB), GAg(Hy) and GAg(EG)
absorption intensity of R6G solution decreased by 57.2% for G nanohybrids. The concentration of hybrids in R6G/hybrids
mixed solution is 0.1 mg mL1. (b) Fluorescence emission spectra of
Ag(EG) hybrids, 41.8% for GAg(Hy) hybrids and 6.9% for G
R6G (1  104 M) at 532 nm excitation with and without GAg(EG)
Ag(SB) hybrids. In detail, positively charged R6G molecule can (0.1 g L1).
be adsorbed onto the surface of negatively charged Ag NPs and
GE sheets through the strong electrostatic attraction. It can be
proved in the SERS spectrum of R6G/GAg(EG), the vibrational sheets is a non-ignorable phenomenon to the SERS activity of
band of AgN has been found at ca. 235 cm1 (Fig. 4a), which is GEAg NP nanohybrids (Fig. 3 and 4). In the SERS spectrum of
in line with previous literature of a chemisorptive bond between R6G/GEAg NPs, the typical 'cathedral peaks' around 1360 and
Ag NPs and R6G via the ethylamino group.32 Compared with 1560 cm1 can be found, which may be attributed to the D and
other samples, the GAg(EG) hybrids had a lower degree of G peaks of GE sheets enhanced by Ag NPs. This phenomenon
reduction and larger size of Ag NPs, which can absorb more R6G indicated that the SERS eect of Ag NPs on GE sheets also gave
molecules onto the Ag NPs and GE sheets. Thus, it gave higher an enhancement of Raman signals on R6G molecules. However,
SERS enhancement. In addition, GE sheets have eciently when the concentration decreased below 1  108 M, only the
quenched the uorescence, which also improve the SERS eect SERS spectrum of GE can be detected, suggesting that the SERS
of molecules.8 This can also be supported by uorescence eect of Ag NPs on GE sheets was higher than that on R6G
emission spectra in our system: the intensity of uorescence for molecules (Fig. S6, ESI).
R6G solution (104 M) with GAg(EG) (0.1 g L1) decreased
nearly 155-fold, indicating signicant uorescence quenching
toward R6G (Fig. 5b), which is much higher than the level 2.3. Fabrication of GEAg NP hybrid lm with high SERS
reported previously.24 sensitivity and reproducibility
Therefore, the SERS eects of the GEAg NP nanohybrids on
SERS substrates need to be cheap, robust, reproducible and
R6G probe molecules are very complicated. So far, we have
stable so that they can be stored for longer time between
determined all the possible enhanced Raman signals that exist,
measurements.33 Undoubtedly, the exible SERS substrate is a
including the weak CM-based SERS eect and uorescence
good choice for routine SERS detection. As discussed above, the
quenching by GE sheets, the SERS eect of Ag NPs on GE sheets
GAg(EG) composite has the optimized SERS activity for
and adsorbed molecules. However, in all the SERS eects, we
detection of R6G molecules. Here, the GEAg NP hybrid lm
determined that the EM-based SERS eect of Ag NPs is most
was fabricated by the vacuum ltration of the GAg(EG)
important. We also noted that the SERS eect of Ag NPs on GE
suspension (0.1 g L1, 2 mL), which would be an excellent

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candidate for SERS-based ultrasensitive molecular sensing.

Fig. 6a shows the scanning electron microscopy (SEM) image of
the macrocosmic surface of the GEAg NP (SB) hybrid lm. The
surface of the lm was relatively at and smooth. Very few fold
structures and almost no nanoparticle aggregation were
observed. Fig. 6b and c show the detailed views of the surface of
the GEAg NP (Hy) hybrid lm. Entire surface of the GE lm was
uniformly coated with
35 nm Ag NPs and only a few aggre-
gated nanoparticles were found. Fig. 7a compares the SERS
spectra of R6G molecules with dierent concentrations
obtained from the GEAg NP hybrid lms. For 1  106 M R6G
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molecules, the GEAg NP substrate exhibited obvious SERS

activity, but it was weaker than that of GAg(EG) composites
used as the SERS substrate (the SERS intensity of the CC
stretching peak ca. 1363 cm1 for GEAg NP nanohybrid lms
was 0.44 fold than that of GAg(EG) composites). As discussed
above, we considered that the GEAg NP nanohybrid lms
contained a higher amount of GE than that of composites,
resulting in the Raman signals of probes being covered by the
high Raman signals of GE itself. In addition, when the
concentration decreased below 1  108 M, only the SERS
spectrum of GE could be detected.
In order to improve its SERS sensitivity, the GEAg NP
nanohybrid lm was treated with thermal annealing at a
temperature of 600  C. Fig. 6d shows the SEM image of the
macrocosmic surface of the treated GEAg NP nanohybrid lm.
Compared with the original lm without treatment, the surface
of the treated lm became very rough and many characteristic
folds of GE sheets can be observed because of the severe
Fig. 7 SERS spectra of R6G molecules adsorbed on (a) the original
reducing reaction at the high temperature. From the detailed GEAg NPs nanohybrid lm and (b) lm after thermal annealing
views of the surface in the treated lm, the Ag NPs covered the treatment. R6G concentrations vary from 1  106 M to 1  1012 M.
entire area of GE sheets (Fig. 6e and f). More importantly, the
size of Ag NPs in the lm increased to 50 nm and the inter-
particle distance decreased, resulting in the higher Ag NP annealing treatment has been improved signicantly, which
density in the treated GEAg NP nanohybrid lm. The SERS can be proved from the SERS spectra of R6G molecules with
sensitivity of the GEAg NP nanohybrid lm with thermal dierent concentrations of 106, 108, 109, 1010, 1011 and

Fig. 6 SEM images of (a) the original GEAg NPs nanohybrid lm and (d) with thermal annealing treatment. The detailed views of the original lm
(b and c) and with treatment (e and f).

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1012 M adsorbed on the treated lm (Fig. 7b). The SERS
intensity of the CC stretching peak at ca. 1363 cm1 for the
treated GEAg NP nanohybrid lm was 24.1 fold higher than
that in the lm without treatment (106 M). Although the
Raman intensity of R6G decreased with the R6G concentration
decrease, the SERS sensitivity of the as-prepared treated GEAg
NP nanohybrid lm is so high that the characteristic Raman
bands of R6G collected on the lm exposed to a low 1012 M
concentration can be clearly identied, suggesting the high
SERS sensitivity of the treated GEAg NP nanohybrid lm. The
signicant improvement of SERS sensitivity is attributed to the
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morphology changes of the GEAg NP nanohybrid lm through
the thermal annealing at the high temperature. The rough
surface of the treated GEAg NP nanohybrid lm can absorb
much more R6G molecules, and the larger Ag NP size and
narrower interparticle distance can provide more hot spots,
which can improve the SERS sensitivity.
Fig. 8a shows the SERS spectrum of crystal violet (CV)
molecules (1  108 M) on the GEAg NP nanohybrid lm
treated by thermal annealing and the CV solid was used for
comparison. It can be clearly seen that the Raman signals of CV
were signicantly enhanced by the GEAg NP nanohybrid lm.
The CV molecules exhibit the vibrational bands corresponding
to the aromatic CC stretching modes at 1619, 1585 and
1535 cm1, a N-phenyl stretching mode at 1370 cm1, and
aromatic CH bending modes at 1171, 915, and 809 cm1.7,34 In
Fig. 8 (a) SERS spectrum of CV molecules with 1  108 M concen-
tration adsorbed on the treated GEAg NPs nanohybrid lm. For
comparison, Raman spectrum of CV solid is presented. (b) RSDSERS
graph: SERS spectra of CV collected on the randomly selected
11 places of the lm (top) and the corresponding RSD values curve
(below).
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addition, Raman signals of CV at the range of 100 to 800 cm1
were also detected, while it is dicult to detect them in the
Raman spectrum of CV solid. The results demonstrated that the
as-prepared GEAg NP nanohybrid lm has high SERS sensi-
tivity. It is known that the poor reproducibility of Raman signals
in traditional SERS analysis has been the main obstacle to the
use of SERS as a routine analytical tool, which is also a big
challenge for graphene-based SERS substrates. To test whether
the as-prepared substrates are able to give reproducible SERS
signals, we collected SERS spectra of 1  108 M CV molecules
from 11 randomly selected positions on the as-prepared GEAg
NP nanohybrid lm under identical experimental conditions.
The comparison results are shown in Fig. 8b. The relative
standard deviation (RSD) curve of 11 SERS spectra, which are
used to estimate the reproducibility of SERS signals, is calcu-
lated by the method reported by Zhang et al. in the previous
literature.35 With this method, an overall impression and a
detailed depiction of reproducibility can be provided. RSD
values of signal intensities of all Raman peaks can be looked up
directly. To the treated GEAg NP nanohybrid lm, the average
RSD values of all Raman peaks are observed to be below 0.13,
revealing a good consistency and reproducibility across the
entire area of the lm. Table S3 gives the RSD values corre-
sponding to the major SERS peaks, which means that the major
SERS peaks have a good reproducibility (Table S3, ESI). It can
also be explained by hot spots as described above. The GEAg
NP nanohybrid lm with the treatment by thermal annealing
has the rough surface, large size of Ag NPs and narrow inter-
particle distance, which provides many more hot spots and
dispersed uniformly across the entire area. Thus, this optimized
GEAg NP nanohybrid lm would be demonstrated by a good
reproducibility of SERS signals. Very recently, Wang reported
that the graphene/Ag NP hybrids spin-coated on the Si substrate
has excellent SERS sensitivity and can detect the R6G molecules
with the minimum concentration of 1013 M.36 To further
evaluate the enhancement eciency of this system, we quote a
simple formula,37 EF (ISERS/Ibulk)  (Nbulk/NSERS), and the
details are provided in the ESI. The EF was 1.4  108 for
aromatic CC stretching modes at 1588 cm1. Our GEAg NP
nanohybrid lm, which has a similar SERS sensitivity, has more
potential SERS application because of the good reproducibility,
large-scale and free-standing features, and it would bring a
great convenience to the routine SERS detection.
3. Conclusion
In summary, two aims have been achieved in this paper: (1)
studies on the SERS mechanism of GEAg NP nanohybrids; and
(2) fabrication of large-scale, exible free-standing and high
SERS sensitive GEAg NP nanohybrid lm. In the rst aim, three
GEAg NP nanohybrids (labeled as GAg(SB), GAg(Hy) and G
Ag(EG)) with dierent reducing degrees of GO and sizes of Ag
NPs were synthesized using sodium borohydride, hydrazine and
ethylene glycol as reducers to clarify the SERS mechanism for
detection of R6G molecules. The results showed that the GE
sheets did improve the SERS activity of GEAg NP nanohybrids
due to their high adsorption and strong uorescence quenching
This journal is The Royal Society of Chemistry 2014

Paper
capability and weak CM-based SERS eect. The EM-based eect
of Ag NPs contributes the most to the hybrids' SERS activity on
R6G molecules. Among the three GEAg NP nanohybrids, the
GAg(EG) hybrid has the best SERS activity due to its highest
adsorption ability and the largest Ag NP size. In the second aim,
the large-scale and exible free-standing GEAg NP nanohybrid
lms with high SERS sensitivity were fabricated through the
vacuum ltration of the optimized GAg(EG) composite
suspension followed by thermal annealing. The SERS sensitivity
of the treated lm has been signicantly improved compared
with the original lm because of great morphology changes of
Published on 18 June 2014. Downloaded by Oakland University on 19/10/2014 06:30:46.
the GEAg NP nanohybrid lm during the thermal annealing.
Both the rough surface of the hybrid lm for adsorption of more
R6G molecules and the larger Ag NPs and narrower interparticle
distance which provide more hot spots lead to the high
sensitivity for detection of R6G. The results obtained from this
work demonstrate that the GEAg NP nanohybrids have great
potential applications in routine analysis using SERS.
4. Experimental section
4.1. Chemicals and reagents
GO was synthesized from puried natural graphite bought from
Shanghai Yifan Company with a mean particle size of 1.5 mm
according to the method reported by Hummers and Oeman.38
Analytical-grade reagents, including silver nitrate (AgNO3),
sodium citrate, sodium borohydride, hydrazine, ethylene glycol,
absolute ethanol, R6G and CV were purchased from Shanghai
Chemical Reagent Corporation and used as received.
4.2. Synthesis of GEAg NP nanohybrids
Three dierent kinds of composites were prepared using
sodium borohydride, hydrazine and ethylene glycol as reducers,
according to the modied method of our previous research
work,26,39 which were labeled as GAg(SB), GAg(Hy) and
GAg(EG), correspondingly. The details of stabilization are
presented in the ESI. The pure GE and Ag NPs were fabricated
for control samples according to the modied method of our
previous work, which are also presented in the ESI.
4.3. Fabrication of GEAg NP nanohybrid lms
For lm preparation, the GEAg NP nanohybrids (especially for
GAg(EG) sample) were dispersed in 2 mL de-ionized water with
sonication at a concentration of 0.1 g L1. The dilute suspen-
sion was vacuum ltrated using a mixed cellulose ester
membrane with 25 nm pores (millipore). The lm was allowed
to dry in a vacuum at 30  C. The obtained lm was thermal
reduced in a tube furnace at 600  C under a H2Ar atmosphere
(volume ratio 1/5) for 2 h and cooled naturally to room
temperature.
4.4. Characterization
The crystal structure of the composites was characterized by a
Philips 1730 powder XRD with Cu Ka radiation (l 1.5406 A),
and the diraction data were recorded for 6 min1 between 5
and 80 . The analysis of XPS measurements were performed on
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Journal of Materials Chemistry C
a Thermo ESCALAB 250 spectrophotometer with Al-Ka radia-
tion. TGA was conducted with a Netzsch Sta449C that was tted
to an air gas at a heating rate of 10  C min1. The morphology
and microstructures were observed using a JEOL 2011 TEM at
an accelerating voltage of 200 kV. The surface microstructures
of the lm were observed on a JEOL 6460 SEM equipped with an
EDS. The absorption capacity of composites for R6G molecules
were analyzed by the UV-vis spectra measured on a Shimadzu
UV-2550 spectrophotometer at room temperature. Fluorescence
emission spectra of R6G with and without composites in
ethanol solutions were measured on an Edinburgh Ins FLS920
spectrophotometer.
4.5. SERS measurement
The samples were prepared for Raman measurement according
to the procedure described in the literature.20,40 And the details
are presented in the ESI. The Raman spectra were recorded on
a Thermosher (USA) Raman system with an argon ion laser of
532 nm at the power of 5 mW and laser spot size of 2.1 mm. The
acquisition time for each measurement was 10 s. Replicable
measurements of three times on dierent areas were made to
verify that the spectra were true representations of each exper-
iment. The relative standard deviation is within 5% based on
the strongest peak intensity of R6G and CV on the Raman
spectrum.
Acknowledgements
This work was supported by the National Science Foundation of
China (50902061), Six Personnel Peak Project (2009137) of
Jiangsu Province, and the Scientic and Technical Innovation
Project for Graduate Students (CXZZ12_0670, CXZZ13_0664). YL
and DD acknowledge the nancial support from a WSU start-up
grant.
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