JOURNAL OF RAMAN SPECTROSCOPY

J. Raman Spectrosc. 2007; 38: 1280–1287

Published online 11 June 2007 in Wiley InterScience
(www.interscience.wiley.com) DOI: 10.1002/jrs.1762

Surface enhanced Raman spectroscopic readout on

heavy metal ions based on surface self assembly
Ya-Xian Yuan, Li Ling, Xi-Yu Wang, Mei Wang, Ren-Ao Gu and Jian-Lin Yao∗
Department of Chemistry, Suzhou University, Suzhou 215006, China

Received 17 January 2007; Accepted 12 April 2007

A new method is reported for detecting heavy metal ions by using the self assembled monolayer (SAM)
technique and surface enhanced Raman spectroscopy (SERS). The p-mercaptobenzoic acid (MBA) served
as the SERS readout molecule and the modified tag to attach on the smooth gold substrate as well as the
tag of nanoparticles by the SAM method. Two carboxyl groups from MBA molecules which were attached
respectively to gold substrate and gold nanoparticles were linked through the heavy metal ions (Cu2+ , Pb2+
and Zn2+ ) as bridge, and thus sandwich structure of ‘MBA modified gold substrate/heavy metal ions/MBA
modified gold nanoparticles’ was built for detection. The observation of the oxidation peak of metal
nanoparticles from cyclic voltammetry (CV) curve, gold nanoparticles from scanning electron microscopy
(SEM) images and SERS signal of MBA from the sandwich structure indicated the existence of heavy
metal ions. The difference in the wavenumbers of vibrational modes from MBA in the sandwich structure
constructed by different could be used to identify different heavy metal ions. The assembled structure was
rinsed by strong chelator of EDTA solution to remove the heavy metal ions from the sandwich structure
and thus to obtain a fresh gold substrate modified with MBA for the cyclic detection. Copyright  2007
John Wiley & Sons, Ltd.

KEYWORDS: heavy metal ion; readout; surface enhanced Raman spectroscopy; self assembly; gold nanoparticles

INTRODUCTION some new technologies are applied to the detection of trace

Some divalent heavy metal ions pose significant public
health hazards when they present in living organisms,
drinking water, food, paint and waste water. Even though
the total concentration of heavy metal ions is lower than
parts per million (ppm), their toxicity to human health
cannot be ignored because the heavy metal ions cannot
being biodegradable. As a consequence, the detection
and elimination of heavy metal ions become the most
attractive issue in the environmental science. For the
absolute identification and total concentration assessment,
the methods of choice are inductively-coupled-plasma
(ICP) spectroscopy and atomic absorption spectroscopy.1 – 3
Fluorescence, porphyrin derivatives as complex agents,
array-based sensors, magnetic effects, ion-sensitive field
effect transistors have been employed to detect trace heavy
metal ions successfully in the last several decades.4 – 12
Nevertheless, each method has its own advantages and
disadvantages, and thus a development of a reliable, rapid,
low cost and sensitive method is highly desired. Recently,
Ł Correspondence
to: Jian-Lin Yao, Department of Chemistry,
Suzhou University, Dushu Lake campus, Suzhou Industrial Park,
Suzhou 215123, China. E-mail: jlyao@suda.edu.cn
heavy metal ions. For example the electrochemical stripping,
colormetry assisted by UV absorption and hyper-Rayleigh
scattering (HRS) and quantum contact between a pair of
electrodes separated by an atomic-scale gap provide high
sensitivity for the detection. Among them, the voltammetric-
stripping technique is one of the promising in situ tools
for the detection of heavy metal ions.13 – 16 Normally, in
order to increase the detection limitation, the metal ions are
required to deposit onto the Hg electrode by electrochemical
deposition prior to the anode stripping measurements.
The concentration was deduced by the relationship of
the oxidation current and the deposition time. Kim et al.17
reported a simple colorimetric technique for the detection of
small concentrations of aqueous heavy metal ions, including
toxic metals such as lead, cadmium and mercury. The
modified gold nanoparticles were aggregated in solution by
an ion-induced linkage process in the presence of divalent
heavy metal ions. The change in the color of the solution
measured by UV absorption and the enhancement in the
HRS response revealed the presence and the concentration
of heavy metal ions, particular for the spectroscopic silent
heavy metal ions. Tao and his co-workers reported a simple
method for detecting trace amounts of metal ions by using

Copyright  2007 John Wiley & Sons, Ltd.

 Surface enhanced Raman spectroscopic readout on heavy metal ions based on surface self assembly
electrodes separated by atomic scale gaps. The heavy metal
ions were richened in the atomic scale gaps. By sweeping
the electrode potentials cathodically, metal ions were then
deposited into the gap and form a point contact between
the two electrodes, which triggers a quantum jump in
the conductance. By measuring the relationship of the
conductance of the atomic scale gap and the electrochemical
deposition time, the sensitivity was increased significantly
to detect trace metal ions with amount of about 10 ppt.18
Raman spectroscopy exhibits a huge potential applica-
tion in analytical, surface, biomedicine science because of its
high selectivity and low absorption of aqueous solution.19
The main advantage of Raman spectroscopy is its capability
to provide rich spectral information on the molecular struc-
ture and it can be used under ambient conditions in almost
every environment without the special requirement for the
sample preparation.20 Despite of all these favorable aspects,
conventional Raman spectroscopy has been considered to be
more useful for the structural characterization rather than the
sensitive qualitative or quasi-quantitative analysis for trace
analyte. Furthermore, Raman spectroscopy usually lacks suf-
ficient sensitivity for detecting the signal scattered from
analyte in very low concentration scale because of the small
cross section (typically 10
30 –10
25 cm2 /molecule compared
to 10
20 cm2 /molecule for IR and 10
16 cm2 /molecule for
fluorescence) for Raman scattering which results in lower
sensitivity than IR spectroscopy and fluorescence.21 There-
fore, the ultratrace detection with single molecules was
achieved simply by using fluorescence.22,23 However, for
Raman spectroscopy, the high sensitivity can be obtained
under favorable resonance conditions.24 Fortunately, the dis-
covery of surface enhanced Raman spectroscopy (SERS) in
which the surface Raman signal was enhanced by four to six
orders at rough coinage metal surface with nanometer scale
structure, provided an alternative method for the detection
of trace analyte because of its high surface sensitivity.25 – 30
Moreover, as a spectroscopic method, SERS could combine
the merits of fluorescence and Raman spectroscopy, i.e. the
high sensitivity and rich structural information. Most excit-
ing, a dramatic SERS effect with 14 orders magnitude was
observed from the molecule attached to the metal nanoparti-
cles (e.g. gold and silver colloids with a favorable size). The
SERS effect yielded from the nanoparticles with a certain
diameter was comparable to or even exceeded those from
fluorescence. Therefore, the discrepancy between Raman
scattering cross sections and fluorescence cross section can
be totally overcome, and thus real single molecule detection
was achieved by SERS on silver nanoparticles.11,19,20 This
breakthrough has led to renewed interest in the exploration
of this information-rich spectroscopy as a promising tool for
rapid, low-level readout in analytical science.
In the present studies, SERS is explored as a sensitive
detection method for the heavy metal ions. Smooth gold
surface modified with p-mercaptobenzoic acid (MBA) served
as the substrate (sensor) for capturing heavy metal ions
Copyright  2007 John Wiley & Sons, Ltd.
1281
and the MBA modified gold nanoparticles, and thus a
sandwich structure of ‘modified gold nanoparticles/heavy
metal ions/modified gold substrate’ was built through the
carboxylate linkage of heavy metal ions for the detection.
Cyclic voltammetric (CV), scanning electron microscopy
(SEM) and SERS were employed to characterize the sandwich
structure. The sandwich structure was broken by the
assistance of the strong chelation of EDTA with heavy metal
ions, and thus the modified substrate can be recycled for the
detection.
EXPERIMENTAL
Synthesis and modification of gold nanoparticles
To prepare the aqueous gold nanoparticles, 0.6 ml of
10 mg/ml trisodium citrate was added to 100 ml of
0.1 mg/l boiling tetrachloroauric acid aqueous solution
while stirring.31,32 The mixture was kept boiling for about
15 min until a deep wine red color was observed, indicat-
ing the formation of gold nanoparticles, and then cooled
down to room temperature. The gold nanoparticles were
preconcentrated before modification. Transmission electron
microscopy (TEM) measurements showed that the mean
diameter of the prepared gold nanoparticles was about
50 nm. For the preparation of MBA modified gold nanopar-
ticles, 1 µl of 1 mmol/dm3 MBA ethanol solution was added
to 1 ml preconcentrated gold nanoparticles solution to full
cover nanoparticles with MBA monolayer.
Self-Assembled Monolayer (SAM) Formation of
MBA on gold surface
Thin gold film evaporated on a glass slide with an ultrathin
chromium layer was used as substrate. The film was first
rinsed by ethanol twice and dried by nitrogen. After that,
it was annealed by gas light for about 1 min to form a
large terrace with an atomic scale flat, and cooled down
to room temperature in the nitrogen environment. The
formation of the self-assembled monolayer (SAM) of MBA
was accomplished by immersing the gold substrates into
1 mmol/dm3 MBA solution for about 4 h, and rinsed by
ethanol twice and Mill-Q water to remove the physical-
adsorbed MBA molecules.
Immobilization of metal ions and MBA capped
gold nanoparticles onto gold substrate
The MBA modified gold substrate was immersed into copper
nitrate solution for about half an hour for capturing the
copper ion by the terminated carboxyl group on the surface,
and cleaned by three distilled water three times. At last, the
MBA modified gold substrate linked with copper ion was
exposed to MBA capped gold nanoparticles basic solution
for about half an hour to attach the gold nanoparticles
on to the substrates.33 – 35 After being taken out the gold
nanoparticles solution, the substrate was rinsed by Mill-Q
water several times to remove any nanoparticles which was
J. Raman Spectrosc. 2007; 38: 1280–1287
DOI: 10.1002/jrs
1282 Y.-X. Yuan et al.
not bonded to the gold surface. The modified gold substrates
were measured by electrochemical CV (CVs were recorded
on a CHI660 electrochemical workstation, USA), SEM (SEM
Hitachi S-570 Japan) and SERS (performed on a LabRam
HR800 confocal micro Raman system with the excitation line
of 632.8 nm, Dilor, France). The solution was prepared by
fresh Mill-Q water for avoiding the influence the trace metal
ions from the air.
RESULTS AND DISCUSSION
The principle of the detection
Scheme 1 describes the construction on the sensor for
detection of heavy metal ions in four steps. In the present
case, the copper (Cu2C ), zinc (Zn2C ) and lead (Pb2C ) ions
were selected as examples to prove the reliability of this
methodology.33 – 35 MBA was used to immobilize heavy
metal ions as well as the reporter molecule for the SERS
readout because of its relative simple structure and strong
Raman scattering signal from gold nanoparticles. Four steps
involved in the construction of the sandwich structure
as: (i) the formation of MBA SAM onto an atomic flat
gold film (nonSERS active) on glass surface (shown as
step A), (ii) capturing copper ions onto the MBA modified
substrate from the analytical medium (shown as step B),
(iii) immobilizing MBA capped gold nanoparticles through
the copper ions carboxylate linkage (step C), (iv) recycling
the sensor by removing copper ions by strong chelator
(step D). In the step A, the MBA was adsorbed onto
gold surface through its S atom to form a compact MBA
monolayer terminated with a free COOH group. The Raman
spectroscopy measurements showed that the MBA Raman
signal can not be observed on the smooth gold substrate.
To metalate the MBA modified gold surfaces, the substrate
was immersed into 0.01 mol/dm3 copper nitrate solution.
By the copper ions carboxylated linkage chemistry, copper
ions were then attached onto the MBA SAM film, and
hence divalent copper ion left the site to capture another
molecule with carboxyl group (as shown in Scheme 1 step
B). Therefore, copper ions played as the bridge for connecting
other carboxyl groups or capturing other solid surface
terminated with carboxyl groups, for example, carboxyl
groups from the MBA capped gold nanoparticles, and thus
resulting in the formation of sandwich structure of ‘MBA
capped gold nanoparticles/copper ions/MBA modified gold
substrate’. It can be used as the sensor not only for the
detection, but also for enriching the toxic metal ions from the
medium. Therefore, the MBA modified gold substrate could
capture copper ions from solution for the trace amount
detection by the aid of the high sensitive SERS technology.
On the other hand, the concentration of heavy metal ions
was decreased through this method. As a consequence, it
revealed that organic adlayers with rich carboxyl group
could decrease the mobility and concentration of the toxic
divalent heavy metal ions in solution, and thus it might be a
Copyright  2007 John Wiley & Sons, Ltd.
promising technology not only for protecting environments,
but also for the detection of toxic heavy metal ions. For such
sensor, the key to the detection of heavy metal ions is focused
on the signal readout from reporter with ultralow amount. It
is well known that noble nanoparticles with an appropriate
size or the junction between nanoparticles could produce a
giant SERS effect. Furthermore, the previous studies showed
that the gold substrate surface contributed to a significant
localized field enhancement because of the surface plasma
polarization for MBA molecules located in the sandwich
structure, resulting in its large SERS intensities, it was so
called as the coupling effect of localized plasma polarization
and surface plasma polarization between nanoparticles and
a smooth gold surface.36,37 Very recently, Orendorff et al.37
reported that the surface enhancement factor of MBA is
about 107 –109 dependent on the shape of the nanoparticles
between the nanoparticles and a planar surface. Therefore,
the SERS effect form the gold nanoparticles and the plasma
coupling effect between the nanoparticles and gold substrate
enabled us to detect the SERS signal from the reporter
molecules located between substrate and nanoparticles. It
might be a suitable high sensitive sensor for the heavy metal
ion detection.
Electrochemical and SEM detection
So far, there are several methods to characterize/readout the
above mentioned sandwich structure/sensor. For example,
the heavy metal ions could be detected by the electrochem-
ical method based on the CV and appropriate conductivity
of the modified molecules between the metal ions and gold
surface. However, the concentration of heavy metal ions
was quite low on the MBA modified gold surface (normally
8 c
6
Current/µA
4
b
2
a
0
-2
0.0 0.2 0.4 0.6 0.8
E/V (vs. SCE)
Figure 1. Cyclic voltammetric curves of MBA modified gold
electrode immersed into the solution including Cu2C and
followed with an ultrasonic cleaning (a), MBA modified gold
surface immersed into solution without Cu2C and then
immersed into MBA modified silver nanoparticles solution (b)
and sandwich structure electrode as described in text (c) in
0.5 mol/dm3 H2 SO4 .
J. Raman Spectrosc. 2007; 38: 1280–1287
DOI: 10.1002/jrs
 Surface enhanced Raman spectroscopic readout on heavy metal ions based on surface self assembly
less than one monolayer), which resulted in the difficulty
in observation the reduction peak of the metal ions in the
sandwich structure in the CV. Nevertheless, it was worth to
detect electrochemical oxidation of the assembled nanopar-
ticles through the linkage of copper ions by CV technique.
Figure 1 presented the CV curves of MBA modified gold sub-
strate with/without silver nanoparticles. It should be pointed
out that the MBA modified silver nanoparticles severed as
outerlayer of the sandwich structure, in order to avoid the
overwhelming on the oxidation current from gold substrate.
Firstly, the MBA modified gold surface was immersed into
the Cu2C solution with the concentration of 0.01 mol/dm3
and followed with an ultrasonic cleaning and rinsed with
three-times distilled water to remove the physical adsorbed
Cu2C (the surface structure was like C in Scheme 1). It can
be found that no oxidation/reduction peaks were observed,
which indicated that the amount of anchored Cu2C was too
low to be detected by CV (normally, it was less than one
monolayer). Secondly, the MBA modified gold surface was
immersed into pure water without heavy metal ions and then
immersed into MBA modified silver nanoparticles solution.
The CV of such gold surface is presented as Fig. 1(b). Without
the bridge of Cu2C , no sandwich structure was constructed
successfully. As a consequence, no silver nanoparticles were
assembled onto the gold surface. Lastly, the gold surface
was modified as described in Scheme 1, one oxidation peak
located at about 0.55 V was observed in the CV, which could
be assigned to the oxidation of silver nanoparticles (Fig. 1(c)).
Au
A
Glass
Glass
SERS
D
hv
Au
Glass
Cu2+, Zn2+ or Pb2+
Au or Ag nanoparticle
Scheme 1. Schematic diagram of surface enhanced Raman spectroscopy based detection on heavy metal ion. This figure is available
in colour online at www.interscience.wiley.com/journal/jrs.
Copyright  2007 John Wiley & Sons, Ltd.
1283
Therefore, the observation of oxidation of silver nanoparti-
cles indicated the existence of Cu2C , and CV might a rapid
method to detect the existence of heavy metal ions.
However, SEM imaging can provide direct information
on the morphology of the solid surface and gives direct evi-
dence for proving the existence of heavy metal ions. Figure 2
presents the SEM images of the sandwich structures formed
by the carboxylate linkage of Cu2C with different concentra-
tion and Zn2C or Pb2C , respectively. It was unambiguous to
observe that MBA capped gold nanoparticles were assem-
bled onto gold substrate. Moreover, it can be seen from
Fig. 2(a) and (b) that the gold nanoparticles were attached
onto the surface by a monolayer and the density of nanopar-
ticles was dependent on the concentration of copper ions.
The lower of the concentration of copper ions resulted in
the smaller of the nanoparticles density on the gold surface.
Figure 2(c) and (d) shows that Zn2C or Pb2C ions could serve
as the linkage for capturing the gold nanoparticles onto the
MBA modified gold substrate for detection as well. There-
fore, the SEM results can provide direct evidence to prove the
above concept for the construction of the sensor. However,
the SEM measurement is a lab-based characterizing tech-
nique, it was restricted to be a rapid readout method because
its special requirement on the sample preparation and vac-
uum environment for measurements, while for the SERS, it
exhibited ultrahigh sensitivity to single molecule, it does not
require special preparation on sample and it can be made by a
Au
Glass
B
Au
C
Glass
_p-mercaptobenzoic acid (MBA)
J. Raman Spectrosc. 2007; 38: 1280–1287
DOI: 10.1002/jrs
1284 Y.-X. Yuan et al.
a b
300 nm
c d
300 nm
Figure 2. SEM images of gold nanoparticles attached onto MBA modified gold surface by the chelation of metal ions with the
concentration of 0.01 mol/dm3 Cu2C (a), 10
6 mol/dm3 Cu2C (b), 10
6 mol/dm3 Zn2C (c) and 10
6 mol/dm3 Pb2C (d).
portable Raman spectrometer. Therefore, it might be a poten-
tial rapid and high sensitive method for the sensor readout.
SERS detection
The SERS characterization is shown in Fig. 3 in the spectral
region between 900 and 1800 cm
1 together with the normal
Raman spectrum of the MBA solid. The normal Raman
spectrum and the SERS spectra were comparable to the
previous studies on MBA in solution and adsorbed onto
SERS active substrates. Two characteristic strong bands
at about 1589 cm
1 and 1079 cm
1 were observed in the
SERS spectra from the sandwich structure, which were
comparable to the normal Raman spectrum of MBA solid.
These two bands were assigned to
8a and
12 aromatic
ring breathing vibrational mode. Other relative intense
modes, including the CH deformation, the COO
symmetric
stretching mode (normally at about 1350 cm
1 ), are observed
in the SERS spectrum. The observation of
sCOO
in the SERS
spectrum indicates the presence of deprotonated MBA in the
sandwich architecture. As expected, no SH stretching mode
was observed in the SERS spectrum, indicating the MBA
adsorbed on to gold substrate/nanoparticles through the S
atom. On the basis of the surface selection rules, the strong
ring breathing modes in the SERS spectrum showed that
MBA was adsorbed onto gold surface by vertical orientation.
Without the copper ions chelation step, the MBA
capped gold nanoparticles would not attach onto the gold
Copyright  2007 John Wiley & Sons, Ltd.
200 nm
300 nm
substrate, and thus the sandwich architecture was not built
successfully. Therefore, no SERS signal of MBA was detected
for such structure because there were no nanoparticles to
produce SERS effect for the readout of the reporter molecule
(Fig. 3(b)), indicating no heavy metal ions in the analyte.
It is reasonable to assume that the gold nanoparticles
played an important role in the generation of the SERS
effect. It is well known that noble metal nanoparticles with
appropriate size can contribute large enhancement factors
for the SERS effect and even single molecule detection
by SERS has been reported for molecules adsorbed at the
junction site between nanoparticles or some ‘hot particles’
because of the localized surface plasma coupling between
nanoparticles. However, in the present case, the reporter
molecules were anchored between the nanoparticles and
the gold substrate. For such structure, the surface plasmon
polariton from the gold substrate attributed significantly
on the SERS effect of molecules between the substrate
and gold nanoparticles. As a consequence, the plasmon
coupling between the gold nanoparticles and gold substrate
induced giant electromagnetic (EM) enhancement for the
MBA molecules in the sandwich architecture, and the
detection sensitivity is increased dramatically.37 Moreover,
the underneath gold substrate and the gold nanoparticles
were full-covered by MBA molecules, the amount of reporter
molecules for SERS readout were increased. Although the
concentration of the heavy metal ions was quite low,
J. Raman Spectrosc. 2007; 38: 1280–1287
DOI: 10.1002/jrs
 Surface enhanced Raman spectroscopic readout on heavy metal ions based on surface self assembly
the reporter molecules from the metal ions linked gold
nanoparticles and the gold substrate was enough to produce
an observable SERS signal and hence the indirect detection
sensitivity was increased. Therefore, the reporter molecule
SERS readout methodology driven by the copper ions
enables us to detect the heavy metal ions with ultra-low
concentration.
In order to examine the generalization of the above
method, the Zn2C and Pb2C with different concentration
were detected. Figures 4 and 5 present the SER spectra
from the gold substrate with Zn2C or Pb2C , respectively.
By comparing the SER spectra carefully, several key spectral
features should be pointed out as below: (i) the wavenumbers
of the main bands from MBA are different from the Cu2C ,
Zn2C and Pb2C ions. The wavenumber of
8a is up-shifted
with the increase of atomic numbers of metal ions, i.e.
1589 cm
1 for Cu2C , 1590 cm
1 for Zn2C and 1594 cm
1 for
Pb2C , respectively. This is considered to be mainly due to
the difference in the coordination surrounding between the
different metal ions and MBA. The capability to detect the
difference in the wavenumbers of the main bands showed
the advantage of Raman spectroscopy over electrochemistry
and SEM. It might become the key issue for increasing the
2000 2500
300 nm
1589
1079
Raman Intensity
1099
1596
1000 1200 1400
Wavenumber/cm-1
Figure 3. Normal Raman spectrum of MBA solid (a), Raman
spectrum from the gold substrate without copper ions linkage
underwent the immersion in the MBA capped gold
nanoparticles solution (b), SERS spectrum of MBA from the
gold substrate underwent two steps of copper ions linkage (c),
the insert image presented the sandwich structure constructed
by 0.05 mol/dm3 Cu2C .
Copyright  2007 John Wiley & Sons, Ltd.
1285
1590
1079
e
Raman Intensity
d
c
b
a
1000 1200 1400 1600
Wavenumber/cm-1
Figure 4. SERS spectra from the gold substrate without metal
ions linkage (a), with Zn2C of 10
6 mol/dm3 (b), 10
5 mol/dm3
(c), 10
4 mol/dm3 (d) and 10
3 mol/dm3 (e).
c
selectivity of the SERS based method, and thus one can
assume the recognition would be carried out by comparing
a the change of the wavenumber of
8a . Unfortunately, the
slight change of the wavenumber resulted in the difficulties
3000 for recognizing the metal ions from the mixture solution
with several metal ions, (ii) for the same metal ion, the
wavenumbers of MBA did not exhibit any change with the
concentration of metal ions, therefore, based on the above
two features, the possibility for recognizing the kind of
metal ions is reasonable by comparing the change of the
c wavenumbers from the sandwich structure assembled by
different metal ions, (iii) the intensities of main SERS bands
b changed with the concentration of metal ions, it was well
known that the SERS intensity was contributed by several
factors, such as the surface structure, the coverage, the
excitation line, and the complex enhancement mechanism(s),
as a consequence, SERS was considered as a qualitative or
a quasi-quantitative rather than a quantitative method since its
discovery. In the present case, the change in SERS intensity
was frozen after the concentration of metal ions was higher
1600
than 10
5 mol/dm3 (Figs 4 and 5). The SERS intensity was
saturated due to the full coverage of gold nanoparticles
assembled by the metal ions. However, it showed the
dependence on the concentration low as 10
6 mol/dm3 ,
in which the intensity was reduced with the decrease in
the concentration. Therefore, the detection range based on
SERS intensity can be divided to two parts, for the relative
high concentration, SERS served as a qualitative method for
recognizing the kinds of metal ions in the analyte, while
J. Raman Spectrosc. 2007; 38: 1280–1287
DOI: 10.1002/jrs
1286 Y.-X. Yuan et al.

it gives the qualitative and quasi-quantitative information

together when the total concentration of heavy metal ions a
low than 10
6 mol/dm3 .

The regeneration of the substrate for detection

For the economization and practicability, the possibility for
regeneration the detection processes should be considered
as a key issue for the present method, in particular for
the MBA modified gold substrate. The sandwich structure
was constructed by the metal ions linkage between the
MBA modified gold substrate and MBA attached gold
nanoparticles, thus the removal of metal ions would result
in the break of the sandwich structure. As a consequence,
the MBA modified gold substrate left for the next assembly
and detection. Figure 6(a) shows the sandwich structure for b
the first assembly by 10
4 mol/dm3 Pb2C . After the SERS
detection, the substrate was immersed into EDTA solution
for 2 h, no gold nanoparticle was observed in the SEM image
with large size (Fig. 6(b)). This mainly due to the metal
ions together with the MBA capped gold nanoparticles
were removed by the stronger chelation of EDTA with
metal ions. The refreshed substrate was assembled the
gold nanoparticles as the procedure B and C of Scheme
1 by 10
4 mol/dm3 Pb2C again. Figure 6(c) presents the SEM
image of the sandwich structure, which indicated the MBA
modified substrate could be regenerated by EDTA for the c
cyclic detection.

1081 1594

f
Raman Intensity

Figure 6. SEM images of gold surface with gold nanoparticles

e linked by 10
4 mol/dm3 Pb2C (a), after the above sandwich
structure was immersed into EDTA solution (b) and assembled
the sandwich structure with 10
4 mol/dm3 Pb2C repeatedly (c).
d

CONCLUSION
c
b In summary, the sandwich structure of ‘MBA modified gold
a substrate/heavy metal ions/MBA modified gold nanoparti-
cles’ was built through the copper ions linkage of carboxyl
1000 1200 1400 1600
group from the terminal of gold substrate and gold nanopar-
Wavenumber/cm-1
ticles. It was used to detect the heavy metal ions by the
Figure 5. SERS spectra from the gold substrate with the high sensitive SERS readout technology. The observation of
carboxyl linkage of Pb2C of 10
12 mol/dm3 (a), 10
8 mol/dm3 gold nanoparticles from the CV curves, SEM images and
(b), 10
7 mol/dm3 (c), 10
6 mol/dm3 (d), 10
5 mol/dm3 (e) and SERS signal of MBA indicated the existence of heavy metal
10
4 mol/dm3 (f). ions. The SERS technique could give the qualitative and

Copyright  2007 John Wiley & Sons, Ltd. J. Raman Spectrosc. 2007; 38: 1280–1287
DOI: 10.1002/jrs
 Surface enhanced Raman spectroscopic readout on heavy metal ions based on surface self assembly
quasi-quantitative information together, which showed the
advantages over the electrochemistry and SEM techniques.
Moreover, the sandwich structure was rinsed by strong
chelator of EDTA solution to chelate the heavy metal ions
from the sandwich structure and thus to obtain a fresh MBA
modified gold substrate for the cyclic detection. Although
more issues about this methodology must be addressed,
including the high selectivity, sensitivity, regeneration of the
sensor, the preliminary experiment has shown that SERS
could be developed into a promising tool for detecting the
trace amount heavy metal ions by applying the appropriate
assembling ligand and reporter molecule readout strategy.
Acknowledgements
The authors gratefully acknowledge the financial support from
the Nature Science Foundation of China (20503019, 20573076) and
Natural Science Foundation of Jiangsu Province (BK2005032). YXY
would thank for the partially supported by key academic discipline
of organic chemistry of Jiangsu Province and Suzhou university for
young teachers.
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