Analytica Chimica Acta xxx (xxxx) xxx

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Analytica Chimica Acta

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

Ultrasensitive and recyclable superstructure of AueSiO2@Ag wire for

surface-enhanced Raman scattering detection of thiocyanate in urine
and human serum
Bright Ankudze, Anish Philip, Tuula T. Pakkanen*
Department of Chemistry, University of Eastern Finland, P.O. Box 111, F1-80101, Joensuu, Finland

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

The synthesis of a superstructure of

AueSiO2@Ag wires is reported.

The substrate is active for SERS-based
detection of thiocyanate in different
media.

The substrate can be recycled for
multiple detection of thiocyanate in
different media.

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

Article history: Thiocyanate level in the human system can serve as a biomarker to distinguish smokers from non-
Received 14 August 2018 smokers. Thiocyanate is a potential goitrogen, thus an accurate determination may help to identify
Received in revised form lactating mothers with high thiocyanate dosage, thereby preventing the transport of excess SCN to
12 October 2018
infants. Surface-enhanced Raman spectroscopy has become a versatile and reliable technique to detect
Accepted 18 October 2018
Available online xxx
SCN in different media. However, the conventional surface-enhanced Raman scattering (SERS) sub-
strates used to detect SCN are often discarded after use. The frequent disposal of such metal nano-
particles is detrimental to the environment and makes the SERS-based detection of SCN uneconomical.
Keywords:
Sodium thiocyanate
In this study, we present fabrication of a new, ultrasensitive and recyclable SERS substrate, based on an
Gold nanoparticles AueSiO2@Ag wire (W) superstructure, to detect SCN in different media. The hierarchical AueSiO2@AgW
Surface-enhanced Raman scattering substrate was obtained by forming nano-sized patches of SiO2 on micron-sized AgW and anchoring
Silver wires 30 nm-sized gold nanoparticles on the patches with mercaptopropyltrimethoxysilane. This ultrasensitive
Silica substrate could detect SCN at a low concentration of 0.001 mM in water, and 0.01 mM in urine and
Ultraviolet light human serum. In addition, a facile procedure to regenerate and recycle the SCN bound AueSiO2@AgW
platform in different media has been demonstrated. The insight gained in the present study can serve as
a promising and powerful method for fabrication of active and recyclable substrates for SERS-based
detection of SCN.
© 2018 The Author(s). Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND
license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

1. Introduction

* Corresponding author.
Thiocyanate (SCN) is a complex anion, which can be derived
E-mail address: Tuula.Pakkanen@uef.fi (T.T. Pakkanen). from both organic and inorganic sources [1]. A typical organic

https://doi.org/10.1016/j.aca.2018.10.040
0003-2670/© 2018 The Author(s). Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/
).

Please cite this article as: B. Ankudze et al., Ultrasensitive and recyclable superstructure of AueSiO2@Ag wire for surface-enhanced Raman
scattering detection of thiocyanate in urine and human serum, Analytica Chimica Acta, https://doi.org/10.1016/j.aca.2018.10.040
2 B. Ankudze et al. / Analytica Chimica Acta xxx (xxxx) xxx
source is mustard oil [2], and in inorganic medium, it commonly
exists with sodium and potassium counter ions. The body obtains
SCN from consumable plants that are rich in cyanogenic glyco-
sides, such as cassava, yam, maize, sorghum, cabbage, turnip,
almond and cauliflower [3,4]. Residual cyanide in cigarettes smokes
can also be metabolized into SCN [5,6]. Thiocyanate plays a sig-
nificant role in a lactoperoxidase system (antibacterial activity in
milk), and thus is used as milk supplements [7]. It inhibits the
oxidative damage of cell, thereby acting as an antioxidant [8].
However, when present in excess, SCN becomes detrimental to the
host. SCN inhibits the sodium/iodide symporter (NIS) in the up-
take of iodide by the thyroid gland [9]. This leads to a decrease in
the production of iodine-rich thyroid hormones, triiodothyroxine
(T3) and thyroxine (T4), which control metabolic activities [9]. The
deficiency of these hormones can result into the enlargement of
thyroid tissues, and to a disease known as goiter [9].
Thiocyanate in the body can be found in extracellular fluids such
as saliva, plasma, nasal lining fluid, tears and gastric juices [10e12].
Accurate assay of SCN in these media is imperative to distinguish
smokers from non-smokers, and to help prevent the passage of
excess SCN through breast-feeding to babies. Several analytical
techniques, such as spectrophotometry [13], gas chromatography-
mass spectrometry [14], calorimetry [15], capillary electropho-
resis [16], and high-performance liquid chromatography (HPLC)
[17], have been extensively used for the SCN determination.
However, some of these techniques require complex and expensive
instruments, and are time consuming, while others require the use
of chemicals that can complicate the determination and interfere
with the result.
In recent years, surface-enhanced Raman spectroscopy has been
found to be a simple, less time consuming, highly sensitive and
chemically specific technique, thus capable of providing fingerprint
information about a molecule [18]. The surface-enhanced Raman
scattering (SERS) effect is observed when a molecule, which is
placed on a roughened metal nanostructure, is radiated with a
suitable laser beam [19]. In principle, localized surface plasmon
resonance (LSPR) is excited due to coherent oscillation of conduc-
tion electrons. These LSPRs induce a strong electromagnetic field on
the surface of the metal nanostructure [20]. The field is more
intense at the junctions of nanoparticle assembly (hot spots) due to
the plasmon coupling in these regions, and the Raman mode of a
sample molecule placed at this area is significantly amplified [21].
The effect can also be observed from charge transfer between the
adsorbed molecule and the metal nanoparticle [22]. Therefore,
fabrication of a highly sensitive SERS substrate requires a careful
control of parameters such as the size, density, and inter-particle
distance of the metal nanoparticles [20].
In the past few years, different nanostructured architectures
such as silver aggregates [7], graphene coated gold nanoparticles
[23], starch-reduced gold nanoparticles [18], and gold@silver
nanorods [24] have been demonstrated as active platforms for
SERS-based detection of SCN. However, these substrates had to be
fabricated for every new measurement or analysis. Development of
a simple method to regenerate and recycle thiocyanate-bound SERS
platforms will economize the SERS-based detection process and
reduce the disposal of nanoparticle waste into the environment.
The goal of the present study is to fabricate a new and ultrasensitive
SERS substrate, and to demonstrate the recyclability and reuse of
the new substrate in multiple measurements. Here, silica is directly
grown as patches [25] on the surface of Ag wires (W), and the
SiO2@AgW sample thus formed is functionalized with a thiol
molecule. Finally, gold nanoparticles are bound chemically to the
resulting thiol-functionalized SiO2@AgW (Fig. 1(a)). The obtained
AueSiO2@AgW superstructure served as an active and ultrasensi-
tive platform for the detection of SCN. The SERS properties of the
Please cite this article as: B. Ankudze et al., Ultrasensitive and recyclable superstructure of AueSiO2@Ag wire for surface-enhanced Raman
scattering detection of thiocyanate in urine and human serum, Analytica Chimica Acta, https://doi.org/10.1016/j.aca.2018.10.040
fabricated AueSiO2@AgW towards SCN are studied in aqueous,
urine and human serum media. The recyclability and reusability of
the ultrasensitive AueSiO2@AgW in the different media are also
demonstrated.
2. Materials and methods
2.1. Chemical
Silver nitrate (AgNO3, Merck, Reg. Ph), ethylene glycol (EG,
Merck 99%) and polyvinylpyrrolidone (PVP, Fluka, Mr ~360,000)
were used in the synthesis of silver wires. Gold(III) chloride trihy-
drate (HAuCl4,3H2O Alfa Aesar,  99%) and L-ascorbic acid (AA,
Aldrich  99%) were used in the synthesis of gold nanoparticles.
Tetraethylorthosilicate (TEOS, Aldrich  99%), ethanol (Altia) and
ammonia solution (NH4OH, Merck 25%) were used in the prepa-
ration of silica patches on Ag wires. The surface of the silica-coated
Ag Wire was functionalized with 3-
mercaptopropyltrimethoxysilane (MPTMS, Aldrich 95%). Sodium
thiocyanate (NaSCN) and 4-aminothiophenol (4-ATP Aldrich  97%)
were used as the analyte and probe molecules in SERS studies.
Potassium carbonate (K2CO3, Aldrich  99%), ammonium sulfate
((NH4)2SO4, Merck Reag Ph Eur), potassium bromide (KBr, Merck),
ammonium chloride (NH4Cl, Merck Reag Ph Eur), sodium dihy-
drogen phosphate monohydrate (NaH2PO4$H2O, Alfa Aesar 99.9%),
synthetic urine (Surine, Aldrich) and human serum (Aldrich) were
used as media for SCN detection. Sodium sulfide (Na2S,
Aldrich  98%) was used in the recycle process. Stock sodium sul-
fide should be handled with care, as exposure to moist air can
release H2S. Ultrapure Milli-Q water (18.4 MU cm1) was used for
all reactions in aqueous solution.
2.2. Synthesis of gold nanoparticles
A stabilizer-free gold nanoparticles were prepared from a
modification of the procedure reported by Andreescu et al. [26].
About 10 ml of 0.15 mM HAuCl4,3H2O was reduced with 0.5 ml of
10 mM of L-ascorbic acid under stirring for 5 min. The gold nano-
particles were collected by centrifugation for a further use.
2.3. Synthesis of silver wires
Silver wires were prepared through a modification of the pro-
cedure by Coskun et al. [27]. Ethylene glycol solutions of AgNO3
(0.08 M) and PVP (0.375 M) were prepared. About 5 ml of ethylene
glycol was heated at 180  C under reflux for 1 h, followed by a
simultaneous addition of 3 ml each of the PVP and AgNO3 solutions.
The addition lasted for 6e9 min and the solution was heated under
reflux for another 1 h. The silver wires were collected by centrifu-
gation, washed three times with 18.4 MU cm1 ultrapure water and
dispersed in 10 ml water.
2.4. Coating of Ag wires with SiO2 (SiO2@AgW)
Different volumes of TEOS were used in a series of experiments
to prepare SiO2@AgW having a variable morphology and thickness
of SiO2 [28]. In a typical reaction, three solutions containing 0.5 ml
of AgWs suspension and diluted with ethanol to a volume of 5 ml
were prepared in erlenmeyer flasks. About 1 ml, 3 ml, and 5 ml of
TEOS were added, respectively. Finally, 500 ml of NH4OH was added
to the resulting solution, followed by 1 h of stirring. The different
batches of SiO2@AgW obtained were collected by centrifugation,
washed three times with ethanol and dispersed in 0.5 ml of
ethanol.
In the functionalization of the SiO2@AgWs [29], an ethanolic
 B. Ankudze et al. / Analytica Chimica Acta xxx (xxxx) xxx 3

solution of 3-mercaptopropyltrimethoxysilane (5%, MPTMS, 50 ml),
and NH4OH (28%, 50 ml) were added to the SiO2@AgWs. After 5 min
of ultrasonication, the mixture was left for 30 min. The MPTMS
functionalized SiO2@AgWs were collected by centrifugation and
washed with ethanol. The obtained MPTMS-functionalized
SiO2@AgWs were finally dispersed in 0.5 ml of ethanol.
2.5. Fabrication of AueSiO2@Ag Wire for SERS
Raman signal was exposed to the detector) were used. The sub-
strates were incubated in the analyte solution for 1 h before SERS
measurements. Model UV Lamp-56, 365 nm (230 V, 0.12 Amps) was
used in the recycling of the AueSiO2@AgW substrate.
2.7. Estimation of the enhancement factor (EF) of the
AueSiO2@AgW

About 1 ml of the MPTMS-functionalized SiO2@AgW suspension
was dried on a clean glass plate. Then, 1 ml of the gold nanoparticle
solution was added. The unbound gold nanoparticles were then
removed by rinsing with water.
2.6. Characterization
The morphology and size of gold nanoparticles, silver wires
(AgWs), SiO2@AgW and AueSiO2@AgW samples were observed
with a Hitachi S-4800 field emission scanning electron microscope
(FE-SEM). An energy dispersive X-ray spectrometer (EDS) operating
on a Noran system six (NSS) software was used to analyze the el-
ements in SiO2@AgW. The plasmonic absorptions of the gold
nanoparticles and silver wires (AgWs) were measured with a Per-
kin Elmer Lambda 900 UV/Vis/NIR spectrophotometer. A Renishaw
invia Raman spectrometer operating on a wire 3.4™ software was
used to record all SERS spectra. In the spectral acquisition, a laser
power of about 3 mW, a 20 objective lens, an excitation laser
wavelength of 785 nm and an exposure time of 10 s (how long the
The enhancement factor (EF) of the AueSiO2@Ag was estimated
with 4-aminothiophenol (4-ATP, the most often used probe mole-
cule) using equation (1) [30],
ISURF :NBULK
EF ¼ (1)
IBULK :NSURF
where ISURF is the intensity of the 2110 cm1 band of 4-
aminothiophenol (4-ATP) recorded on the AueSiO2@AgW, NSURF
is the number of the probe molecule giving rise to ISURF, IBULK is the
intensity of the probe molecule recorded on a clean glass plate, and
NBULK is the number of the probe molecule giving rise to IBULK. NBULK
was calculated from the laser spot (20 mm), penetration depth
(20 mm), the molar mass of (125.19 g/mol) and density (1.17 g/ml) of
4-ATP. NSURF was estimated by dropping 1 ml (10 mM) of 4-ATP on
the substrate and calculating the molecular density in the laser spot
area. The EF of the pristine AueSiO2@AgW was estimated 1.3  107
for 4-aminothiophenol. In the estimation, the 4-ATP molecules
were assumed to assemble uniformly on the surface of the sub-
strate via AueS bonds.

Fig. 1. (a) A schematic illustration for the fabrication of AueSiO2@AgW SERS substrate, (b) SEM image of silver wires with an average diameter of 167 ± 7 nm. STEM images of
SiO2@AgW samples prepared with (c) 1 ml, (d) 3 ml, and (e) 5 ml of TEOS solution, (f) surfactant-free gold nanoparticles with an average diameter of 30 ± 4 nm, (g) AueSiO2@AgW
prepared with 1 ml of TEOS. (h) STEM image, EDS maps and spectrum of SiO2@AgW prepared with 1 ml of TEOS, indicating the presence of Si, O, and Ag in the structure. (For
interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Please cite this article as: B. Ankudze et al., Ultrasensitive and recyclable superstructure of AueSiO2@Ag wire for surface-enhanced Raman
scattering detection of thiocyanate in urine and human serum, Analytica Chimica Acta, https://doi.org/10.1016/j.aca.2018.10.040
4 B. Ankudze et al. / Analytica Chimica Acta xxx (xxxx) xxx

2.8. Regeneration of AueSiO2@AgW substrates
The thiocyanate-bound AueSiO2@AgWs substrates were incu-
bated in a 0.01 M Na2S solution for 20 min at ambient temperature
to replace thiocyanate ions with S2. The stock sodium sulfide so-
lution should be handled with care as exposure to moist air can
release H2S, which is a poisonous and flammable gas. Therefore, all
regeneration experiments should be carried out in a well-
ventilated fume hood. The substrates were then removed, washed
with ultrapure water and dried under ambient conditions. The
substrates were finally exposed to ultraviolet radiation
(l ¼ 365 nm) overnight leading to desorption of S2 and recovery of
the AueSiO2@AgWs.
3. Results and discussion
3.1. Design and fabrication of AueSiO2@Ag wire superstructure
According to Fig. 1(b), silver wires having a pentagonal cross
section were obtained by the polyol method [27,31]. In the syn-
thesis of the silver wires, ethylene glycol at a high temperature
(180  C) was used as the reductant of silver ions in the presence of
polyvinylpyrrolidone (PVP). Upon reduction, the PVP selectively
passivates some of the facets of the silver core, the (100) plane is
passivated by PVP leaving the (111) plane to grow anisotropically in
the [110] direction [31]. The growth in the [110] direction results in
the formation of silver wires [31]. As illustrated in Fig. 1(b), the
average diameter and length of the as-prepared silver wires are
167 ± 7 nm and 40 mm, respectively. The plasmonic absorption
maximum of the wires was observed around 402 nm in the visible
region of the electromagnetic spectrum (Fig. S1). The X-ray
diffraction pattern (Fig. S2) of the prepared silver wires, with 2q
angles of 38 , 44 , 64 , 77 and 81, which can be assigned to the
reflections from the (111), (200), (311) and (222) planes, suggests a
well-defined face-centered cubic (FCC) structure.
The AgWs were coated with silica through hydrolysis and
condensation of tetraethylorthosilicate (TEOS) [28]. The STEM im-
ages of SiO2@AgW prepared with different volumes of TEOS, pre-
sented in Fig. 1(c)e(e), suggest that the morphology and thickness
of SiO2 on AgWs vary with the amount of TEOS added. SiO2 on
AgWs prepared with 1 ml of TEOS grew mostly in hemispherical
patches along the surface of the AgWs. The diameter of the silica
patches ranges from 20 to about 130 nm. In the case of SiO2@AgW
prepared with 3 ml, aside the formation of SiO2 patches on AgWs,
the gap between the individual patches started to be filled. When
the volume of TEOS was further increased to 5 ml, SiO2 starts to coat
the entire surface with a thickness of about 70 nm. An EDS mapping
of the SiO2@AgW in Fig. 1(h) confirms the presence of silicon (Si),
oxygen (O) and silver (Ag) in the structure.
In order to fabricate the AueSiO2@AgW superstructure, the
surface of the SiO2@AgW was first modified with 3-
mercaptopropyltrimethoxysilane (MPTMS). In this process, the
MPTMS binds to SiO2 through the condensation and formation of a
disiloxane linkage (SieOeSi). The free eSH group with a strong
affinity towards metals can easily bind with gold nanoparticles
having a relatively weak capping agent. Surfactant-free gold
nanoparticles with an average diameter of 30 ± 4 nm (Fig. 1(f)) and
a plasmonic absorption around 522 nm (Fig. S3) were prepared and
added onto the MPTMS modified SiO2@AgW to form the Aue-
SiO2@AgW superstructure. Fig. 1(g) shows a STEM image of an
AueSiO2@AgW with the nanoparticles randomly distributed across
the entire surface of the SiO2@AgW. In addition, the particles are
densely packed and are either touching each other or separated by a
few nanometers.

Fig. 2. (a) and (b) SERS spectra of 4-aminothiophenol (1  106 M) and NaSCN (1  106 M) measured on the AueSiO2@AgW substrate. (c) Bar chart of the signal intensities of
NaSCN (1  104 M) measured in the presence of different anions (1  104 M) on the AueSiO2@AgW substrate. Error bars show standard deviations from 10 sets of measurements.
(d) Reproducibility of the thiocyanate band at 2110 cm1 on the AueSiO2@AgW substrate. Ten random signals were recorded for the reproducibility studies.

Please cite this article as: B. Ankudze et al., Ultrasensitive and recyclable superstructure of AueSiO2@Ag wire for surface-enhanced Raman
scattering detection of thiocyanate in urine and human serum, Analytica Chimica Acta, https://doi.org/10.1016/j.aca.2018.10.040
 B. Ankudze et al. / Analytica Chimica Acta xxx (xxxx) xxx 5

3.2. SERS properties of the AueSiO2@Ag wire substrate
The sensitivity of the AueSiO2@AgW substrate was studied with
the 4-aminothiophenol (4-ATP) probe molecule. In Fig. 2(a), the
vibrational modes of 4-ATP at 1077 cm1 and 1591 cm1 recorded
on the AueSiO2@AgW substrate can be attributed to CeS and CeC
stretching modes [32]. The enhancement factor of the substrate
using the same 4-ATP molecule was found to be 1.3  107.
The sensitivity of the AueSiO2@AgW substrates towards a
NaSCN analyte was also studied. The most intense Raman modes of
pure NaSCN solid are normally observed around 750 cm1 and
2069 cm1 [7]. These peaks have been assigned to the CeS and the
eSCN stretching modes, respectively. However, adsorption of
thiocyanate onto noble metal surface causes a shift of the Raman
mode at 2069 cm1. The new band around 2110 cm1 is charac-
teristic of eC≡N stretching, and originates from changes in the
molecular structure from the more stable S¼C¼N- resonance form
to eSeC≡N [33]. As presented in Fig. 2(b), the AueSiO2@AgW
substrate could clearly identify the eC≡N stretching mode, which is
characteristic of thiocyanate at 2110 cm1. Still on the sensitivity,
detection of thiocyanate on the AueSiO2@AgW substrate was also
examined in the presence of different anions. Although it is known
that the detection of thiocyanate can be influenced by the presence
of other anions [18], the AueSiO2@AgW substrate could reliably
detected thiocyanate in the presence of bromide, carbonate, chlo-
ride, phosphate, and sulfate anions (Fig. 2(c)).
In addition to sensitivity, the signal reproducibility of the sub-
strate was also estimated. The determination was done by
recording 10 random spectra of SCN on the substrate and calcu-
lating the relative standard deviation (RSD). The RSD obtained from
the 10 random measurements was ±11%, indicating that the
AueSiO2@AgW substrate has a good signal reproducibility
(Fig. 2(d)).
3.3. Detection of SCN in water, urine and human serum
Fig. 3(a), (b), and (c) show the SERS spectra of NaSCN recorded in
water, 5% human serum, and 5% urine on the AueSiO2@AgW sub-
strate. Different concentrations of the aqueous NaSCN solution
ranging from 100 mM down to 0.001 mM were studied. The limit of
detection (LOD) of thiocyanate in water was 0.001 mM, which is
lower than those recorded with the starch-reduced gold nano-
particles [18], silver aggregates [7], and gold@silver nanorods [24].
In 5% urine and 5% human serum, the limits of detection were the
same 0.01 mM. In Fig. 3(d), the intensity variations of the NaSCN
signal at 2110 cm1 in the three media were compared. Spectra
recorded in water were more intense than those measured in urine
and human serum. The lower intensities may be due to different
interfering species in urine and human serum, which may inhibit
adsorption of SCN on the surface of the AueSiO2@AgW substrate.
It has been reported, based on the statistical data of Tsuge et al. [34]
that the common level of thiocyanate in human blood plasma

Fig. 3. SERS spectra of different concentrations of NaSCN molecules measured in (a) water, (b) 5% human serum, (c) 5% urine, and (d) comparison between the SERS intensities of
NaSCN at the different concentrations in the three media. Error bars in (d) show standard deviations from 10 measurements.

Please cite this article as: B. Ankudze et al., Ultrasensitive and recyclable superstructure of AueSiO2@Ag wire for surface-enhanced Raman
scattering detection of thiocyanate in urine and human serum, Analytica Chimica Acta, https://doi.org/10.1016/j.aca.2018.10.040
6 B. Ankudze et al. / Analytica Chimica Acta xxx (xxxx) xxx

Fig. 4. SERS results of NaSCN (10 mM - 0.1 mM) in water (a)e(b) and 5% urine (c)e(d) measured on AueSiO2@AgW substrates. The corresponding correlations between the intensity
and concentration are shown in (b) and (d). Error bars show standard deviations for 10 measurements.

ranges from 1.7 to 290 mM with an average value of 72.4 ± 77.4 mM.
Therefore, the sensitivity and detection range of the AueSiO2@AgW
substrate are suitable to assay thiocyanate in blood samples.
Fig. 4 shows the SERS results of different concentrations of
NaSCN, ranging from 10 mM to 0.1 mM, measured in water and 5%
urine, as well as, the correlation between the SERS intensity at
2110 cm1 against the concentration. As shown in Fig. 4(b) and (d),
a linear relation between the intensity and concentration was
observed within the concentration range of 0.1 mMe0.9 mM in both
water and urine samples. The equations of the calibration curves
are y ¼ 10793x þ 920.9 (R2 ¼ 0.9728) and y ¼ 1850x þ 905
(R2 ¼ 0.9767) for NaSCN in water and 5% urine, respectively.
3.4. Recyclability of AueSiO2@AgW substrate for multiple detection
of thiocyanate
Recycling in the context of the present study means an effective
removal of SCN adsorbed on the surface of AueSiO2@AgW sub-
strate, so that the substrate can be reused. Studies have shown that
thiol molecules on gold surface can be removed by several tech-
niques such as thermal desorption and oxidation in aqueous solu-
tion [35]. However, an elevated temperature treatment of the SERS
platform may lead to extensive aggregation of gold nanoparticles,
which can adversely affect the performance of the sensor platform.
Oxidation of thiocyanate in an aqueous solution of an acid, on the
other hand, may lead to the formation of HCN [36], which is a
highly toxic gas. Ultraviolet light irradiation has also been reported
as a mild and efficient method to remove thiols from gold surfaces
[35]. However, as demonstrated in the present study (Fig. S4(a)), a
direct irradiation of a SCN bound AueSiO2@AgW sample with the
UV light (l ¼ 365 nm) does not lead to recovery of the substrate as a
new weak broad band appears around 2190 cm1. Therefore, a
straightforward approach to regenerate the thiocyanate-bound
substrate for SERS applications is needed.
Fig. 5(a) illustrates a proposed recycling of the AueSiO2@AgW
platform for a multiple usage. The thiocyanate-bound Aue-
SiO2@AgW substrate is dipped in an aqueous Na2S solution, where
SCN on AueSiO2@AgW substrate is replaced by a S2 anion. The
replacement of the SCN by S2 is monitored with disappearance of
the band around 2110 cm1 (Fig. S4(b)). After the replacement, the
substrate is rendered inactive, and could not be used for further
detection of SCN due to the presence of S2 on the AueSiO2@AgW
substrate. The recovery and reactivation of the substrate, essential
for recycling, are accomplished through desorption of S2 from the
AueSiO2@AgW substrate with ultraviolet (UV) irradiation. It worth
noting, that the UV light treatment does not induce a collapse of the
AueSiO2@AgW superstructure as the exposure only affects the
monolayer of adsorbates present on the AueSiO2@AgW surface.
Two AueSiO2@AgW SERS substrates were prepared and used
for five consecutive regeneration-detection cycles, one in water,
and the other one in 5% urine. In Fig. 5(b)e(e), a comparison made
between the signal intensities of SCN recorded on the fresh and
recycled AueSiO2@AgW substrates reveals that the signal intensity
level on the recycled substrate is lower. The decrease of around 30%
in intensity may result from an incomplete desorption of S2 from
the AueSiO2@AgW substrate during the UV exposure. The rough
surface nature of the AueSiO2@AgW SERS platform involves a
possibility that S2 can adsorb on sites where it is protected from
the UV flux. Such areas may include interstitial sites of a nano-
particle assembly, which are also the “hot spot” regions. Therefore,
the presence of the residual S2 anions on the AueSiO2@AgW
platform may decrease the number of active sites available for
adsorption of the SCN analyte and thus may lower the amount of
SCN adsorbed on the sensor surface compared to the pristine
surface.
After the first regeneration, the SERS signals of thiocyanate
recorded from the five consecutive measurements on the Aue-
SiO2@AgW SERS platforms showed no constant decrease in the

Please cite this article as: B. Ankudze et al., Ultrasensitive and recyclable superstructure of AueSiO2@Ag wire for surface-enhanced Raman
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 B. Ankudze et al. / Analytica Chimica Acta xxx (xxxx) xxx 7

Fig. 5. (a) A schematic illustration of a procedure for recycling of the AueSiO2@AgW substrate. The thiocyanate-bound AueSiO2@AgW substrate on a glass plate is first immersed in
an aqueous solution of Na2S (0.01 M) for 20 min. The substrate is then removed, dried and irradiated with ultraviolet light overnight. Recyclability and reusability of the Aue-
SiO2@AgW in water (b)e(c) and in urine (d)e(e). Error bars show standard deviations from 10 measurements.

signal intensity, signifying a good stability of the regenerated sen-
sors. The relative standard deviation (RSD) values of the signal in-
tensity at 2110 cm1 recorded on the regenerated AueSiO2@AgW
substrates for the five consecutive measurements in water and
urine was ±4% and ±5%, respectively. This indicates that the SERS
signals recorded on the recycled AueSiO2@AgW are repeatable.
Moreover, the regenerated substrate could still detect SCN
molecules at a low concentration of 0.8 mM in water, suggesting that
the recycled AueSiO2@AgW substrates are still capable of low
sensing of SCN. Fig. 6 shows the SERS spectra of different con-
centrations of NaSCN in water and 5% urine measured on the re-
generated AueSiO2@AgW substrate. A linear relation was observed
between the thiocyanate concentration and the SERS intensity at
2110 cm1 within the range of 1e8 mM. The equations of the cali-
bration curves were y ¼ 407.13x þ 723.29 (R2 ¼ 0.9787) and
y ¼ 995.21x þ 7229.3 (R2 ¼ 0.9604) for NaSCN in 5% urine and
water, respectively.
4. Conclusions
In this study, we present the fabrication of a new surface-
enhanced Raman scattering substrate for the detection of a low
concentration of sodium thiocyanate in water, urine and human
serum. Through the assembly of gold nanoparticles, silica and Ag
wires, an ultrasensitive AueSiO2@AgW SERS substrate was formed.
The limits of detecting NaSCN in water, 5% urine and 5% human
serum, using the AueSiO2@AgW SERS platform were 0.001 mM,
0.01 mM, and 0.01 mM, respectively. In addition, we have demon-
strated a straightforward method to regenerate the AueSiO2@AgW
platform for multiple detection of NaSCN. The relative standard
deviation of the SCN intensity around 2110 cm1 in five consec-
utive measurements on the same regenerated substrate was ±4% in

Please cite this article as: B. Ankudze et al., Ultrasensitive and recyclable superstructure of AueSiO2@Ag wire for surface-enhanced Raman
scattering detection of thiocyanate in urine and human serum, Analytica Chimica Acta, https://doi.org/10.1016/j.aca.2018.10.040
8 B. Ankudze et al. / Analytica Chimica Acta xxx (xxxx) xxx

https://doi.org/10.1016/j.aca.2018.10.040.

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Please cite this article as: B. Ankudze et al., Ultrasensitive and recyclable superstructure of AueSiO2@Ag wire for surface-enhanced Raman
scattering detection of thiocyanate in urine and human serum, Analytica Chimica Acta, https://doi.org/10.1016/j.aca.2018.10.040