Research Article

Cite This: ACS Appl. Mater. Interfaces 2019, 11, 28546−28553 www.acsami.org

Carbon Dot-functionalized Interferometric Optical Fiber Sensor for

Detection of Ferric Ions in Biological Samples
Stephanie Hui Kit Yap,† Kok Ken Chan,† Gong Zhang, Swee Chuan Tjin, and Ken-Tye Yong*
School of Electrical and Electronic Engineering, Nanyang Technological University, 50 Nanyang Avenue, Singapore 639798,
Singapore
*
S Supporting Information

ABSTRACT: This work reports an interferometric optical

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microfiber sensor functionalized with nitrogen- and sulfur-

codoped carbon dots (CDs) for the detection of ferric ions
(Fe3+). Compared to other CD-based ferric ion sensors, the
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sensing mechanism of this presented sensor is dependent on

the refractive index modulations due to selective Fe3+
adsorption onto the CD binding sites at the tapered region.
This is the first study in which CD-based sensing was
performed at the solid phase as a chelator, which does not rely
on its fluorescence properties. The detection performance of
the proposed sensor is not only comparable to a conventional
fluorescence-based CD nanoprobe sensor but also capable of delivering quantitative analysis results and ease of translation to a
sensor device for on-site detection. The presented sensor exhibits Fe3+ detection sensitivity of 0.0061 nm/(μg/L) in the linear
detection range between 0 and 300 μg/L and a detection limit of 0.77 μg/L based on the Langmuir isotherm model. Finally, the
potential use of the CD-functionalized optical microfiber sensor in the real environmental and biological Fe3+ monitoring
applications has also been validated in this work.
KEYWORDS: carbon dots, heavy metal ion, optical microfiber sensor, chelator, nanomaterial

1. INTRODUCTION
Iron is geologically abundant and biologically essential to every
living entity.1 It appears as a vital micronutrient in human diets
and commonly exists in its most stable form of ferrous (+2)
and ferric (+3) oxidation states. Aside from its primary role in
the formation of heme, iron plays an important role in the
formation of iron−sulfur clusters, which serve as electron
transfer groups that mediate redox reactions in the biological
system.2 Both deficiency and over-accumulation of iron in the
human body have been reported to elevate the risk for serious
disease conditions such as anemia,3 ischemia, and neuro-
degenerative diseases.4 For these reasons, iron detection in
water, food, and biological fluids has been attracting great
interest from researchers to seek novel functional materials and
systematic iron sensing mechanisms.
Carbon dots (CDs), also known as carbon nanoparticles, are
the latest addition to the carbon-based fluorescent nanoma-
terial family and have been increasingly popular due to their
attractive properties such as facile synthesis, biocompatibility,
good aqueous stability, and unique optical characteristics.5
CDs favor a broad range of applications such as sensing,
cellular imaging, cancer therapeutics, optoelectronics, and
others.6 The potential use of CDs for iron detection has been
demonstrated in many recent works. These include using citric
acid and tris as the precursors to synthesize CDs for ferric ion
(Fe3+) detection with a limit of detection (LOD) of 1.3 μM,7 a
facile preparation of the CD-based Fe3+ nanoprobe sensor
using DL-malic acid as the carbon source with a linear detection
range of 6−200 μM and an LOD of 0.8 μM,8 and the nitrogen-
doped CDs that exhibit fluorescence quenching response
toward Fe3+ over a concentration range of 0−1.6 μM with an
LOD of 0.05 μM.9 Despite the attractive sensing performance,
these nanosensors are generally carried in the liquid phase.
Moreover, these sensors require a bulky and expensive
spectrometer to read the changes in fluorescence emission
intensity, which further hinders the widespread use of CDs for
on-site detection. In regard to these, the CD-based
colorimetric test strip was proposed to overcome these
challenges; however, this solution appears to be more
pronounced for qualitative measurement.10
To circumvent this, optical fibers emerge as an alternative by
offering a wide variety of sensing system designs that include
variants of fiber structures, optical interrogation methods, and
the versatility to incorporate nanomaterial as the transducer
layer to achieve selective and sensitive detection of specific
target molecules. For instance, an optical tapered fiber, or
simply a microfiber, is known for its superior sensitivity for
trace analysis of chemical analytes via the interaction between
the evanescent wave and the target analyte at the receptor-
solute interface.11 On top of that, optical microfibers exhibit
Received: May 22, 2019
Accepted: July 16, 2019
Published: July 16, 2019

© 2019 American Chemical Society 28546 DOI: 10.1021/acsami.9b08934

ACS Appl. Mater. Interfaces 2019, 11, 28546−28553
ACS Applied Materials & Interfaces
great surface chemistry versatility that allows various kinds of
biological or chemical receptors to be coated at the outer
surface of the tapered region. For example, a tapered fiber
platform demonstrated its high degree of flexibility to
functionalize different chelating agents on the tapered region
to target Pb2+, Cu2+, Zn2+, and Cd2+ individually in aqueous
samples.12 Another study implementing a cascade fiber taper
design, which incorporated the multilayer film comprising
chitosan/carbon nanotubes/polyacrylic acid for the detection
of Ni2+, was able to achieve an LOD of 0.3 mM.13 Apart from
the widespread use of an optical microfiber for metal ion
sensing applications, its viability to be translated into a
practical handheld device has also been reported recently,
further increasing the popularity of the optical microfiber-
based sensor.14
Unlike the conventional use of CD-based nanoprobes that
primarily focus on the changes in fluorescence intensity, this
work employed an optical microfiber as a platform to
functionalize nitrogen- and sulfur-codoped CDs onto the
sensing region for selective detection of Fe3+ in both aqueous
and biological environments. The CDs serve as chelators to
induce optical spectrum shifts upon detection of Fe3+ as a
result of the minute refractive index change at the sensing
region. This approach offers a simpler and more economical
experimental setup where a high-resolution charge-coupled
device spectrometer is not required. Moreover, the CD-
functionalized optical-microfiber-based sensor allows a facile
detection protocol and portability of the sensing system for on-
site measurement. The proposed sensor was utilized to
sensitively and selectively detect Fe3+ in aqueous and biological
samples.
2. EXPERIMENTAL SECTION
2.1. Reagents and Materials. Citric acid (>99.5%), thiourea
(>99%), sulfuric acid (H2SO4) (97−99%), hydrogen peroxide
(H2O2) (15%), ethanol (95%), (3-aminopropyl)triethoxysilane
(APTES) (99%), N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide
hydrochloride, EDC, N-hydroxysuccinimide (NHS) (98%), phos-
phate-buffered saline (PBS), and Dulbecco’s modified eagle’s medium
(DMEM) reagent were bought from Sigma-Aldrich. Iron(III)
chloride (98%) was purchased from Acros Organics. Fetal bovine
serum (FBS) was purchased from Thermo Fisher Scientific, and horse
serum was purchased from ATCC. All chemicals were used as
received without further purification. Deionized (DI) water used was
purified by the Milli-Q water purification system.
2.2. Preparation of CDs. The nitrogen- and sulfur-codoped CDs
were prepared according to an earlier report.15 Briefly, 0.5 g of citric
acid and 0.5 g of thiourea were dissolved in 25 mL of DI water to
form a clear homogeneous solution. The solution was then heated up
using a conventional domestic microwave oven at 800 W for 6 min.
The resulting blackish-brown precipitates were then allowed to cool
to room temperature before being added with 30 mL of water. The
mixture was then subjected to ultrasonication in an ultrasonic water
bath for 30 min to fully disperse the carbonized precursors. The
solution was centrifuged at 15 000 rpm for 15 min and filtered using a
0.22 μm membrane filter followed by dialysis against DI water for 24
h to remove unreacted precursors before use.
2.3. Fabrication of Optical Microfiber. The cladding of a single-
mode optical fiber (SMF-28) was partially removed at the center part,
and the fabrication of the optical microfiber was assisted by a Vytran
machine (GPX-3000). The system was preset with a normalized
power filament of 44 W, and the taper profile was fixed at 6 mm of
taper waist length, 3 mm of down-taper length, 2 mm of up-taper
length, and 7.9 μm of taper waist diameter.
2.4. Deposition of CDs onto the Tapered Region. Prior to
functionalizing the as-synthesized CDs onto the tapered region,
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Research Article
microfibers were dipped into piranha solution (3:1 v/v ratio of
concentrated sulfuric acid (H2SO4) and 15% hydrogen peroxide
(H2O2)) at room temperature for 10 min. Following this, the
microfibers were rinsed thoroughly with deionized water and dried
completely. This prepares a hydroxylated surface for the subsequent
interaction with a silane coupling agent, APTES, which was prepared
with ethanol (0.625% v/v). The silanization step was carried out for 1
h at room temperature with continuous stirring at 300 rpm. The
silanized microfibers were then rinsed with ethanol to remove weakly
bonded molecules and dried completely. In the meantime, 0.225 mg/
mL as-synthesized CDs were prepared in phosphate-buffered saline
and the conjugation reactions between the active amine groups on the
tapered region and the surface moieties of the as-synthesized CDs
were performed using the EDC (147.6 μM)−NHS (147.7 μM)
coupling route. Finally, the CD-functionalized microfibers were
removed from the solution, rinsed with deionized water, and left to
air-dry before use.
2.5. Characterization of CDs and CD-Functionalized Micro-
fibers. The transmission electron microscopy (TEM) image was
captured using JEOL 2010UHR, and the size of the CDs was analyzed
using ImageJ. The CD-functionalized optical microfiber’s image,
surface elemental mapping, and analysis were performed using a
scanning electron microscope (SEM, JEOL JSM 6360) and energy-
dispersive X-ray spectroscopy (EDS, JEOL JED-2300). Surface
functional groups on the surface of the as-synthesized CDs and
CD-functionalized optical microfiber were determined using a
Shimadzu IRPrestige-21 attenuated total reflection Fourier transform
infrared spectrophotometer (FT-IR).
2.6. Instrument Setup and Measurement. A broadband
superluminescent light-emitting diode (SLED) (1525−1570 nm)
was used as the incident light source connected to one end of the
microfiber that was secured in place using a pair of magnetic fiber
clamps. Another end of the microfiber was connected to an optical
spectrum analyzer (OSA) for the real-time visualization of the output
transmission spectra. Deionized water was introduced twice to the
tapered region to ensure that the output spectrum is stable. Next, the
sample solution was drop-cast onto the tapered region and left
undisturbed for 4 min, followed by rinsing the microfiber and
introducing deionized water to the tapered region again to obtain
post-metal-binding output spectrum of the microfiber. Finally, the
changes in the spectrum shift were recorded accordingly. All
measurements were performed at ambient temperature ∼24 °C and
in a neutral pH environment, and all CD-functionalized optical
microfibers used in this work were meant for one-time use only.
3. RESULTS AND DISCUSSION
3.1. Sensing Principle and Numerical Analysis. The
Fe3+ sensor was implemented using a CD-functionalized
optical microfiber as the sensing probe. An optical microfiber
is a genre of interferometric optical fiber sensor and can be
fabricated using common single-mode fibers,16 multimode
fibers,17 or even special fibers such as photonic crystal fibers18
and hollow-core fibers.19 In this work, a standard SMF-28 fiber
was used to fabricate the bare microfiber. The overall sensing
system (Figure 1A) comprised a broadband superluminescent
light-emitting diode (SLED), the optical microfiber sensor that
was secured in place on an x-translational stage, and an OSA.
Interference of light occurs by virtue of the optical path length
difference due to waveguide dispersion despite the fixed
physical tapered length.20 Figure 1B depicts that the incident
light from the SLED coupled into the microfiber propagates as
a fundamental mode. At the down-taper region, the
fundamental mode, HE11, was excited to the first dominant
higher-order mode, HE12.21 As the light reaches the up-taper
region, higher-order modes will interfere or couple back into
the fundamental mode, resulting in a modal interference
spectrum that can be expressed as
DOI: 10.1021/acsami.9b08934
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As seen in eqs 5 and 6, Δλ is found to be a function of nmedium

and ρ, which are directly related to the detection sensitivity of
the optical microfiber sensor. The sensitivity of an optical
microfiber is primarily dependent on the interaction between
the evanescent wave and the interface surrounding the tapered
region. Hence, the evanescent wave penetrating further into
the surrounding medium will result in the optical microfiber
being more responsive to small changes that occur at the
surface interface of the tapered region. The penetration depth,
Dp, of the evanescent wave is delineated as the distance away
from the interface at which the magnitude of the electric field
at the surface begins to decay to its 1/e value and is
mathematically described as
λ
Dp = 2
2π (ncladding sin 2 θ ‐nmedium
2
)1/2 (7)

where θ is the angle of incidence at the core−cladding

interface.
Based on the principle above, we simulated the optical
microfiber used in this work using the finite-difference time-
domain method. Figures 2A−D illustrates the simulated
Figure 1. (A) Schematic of the sensing system, (B) sensing principle,
and (C) molecular structure of the as-synthesized CDs.

I = I1 + I2 + 2 I1I2 cos ϕ (1)

where I is the total intensity, I1 and I2 are the intensities of

HE11 and HE12 modes, respectively, and ϕ is the phase
difference between these two modes.
For a uniform beating region, the phase difference is given as
ϕ = ΔβL (2)

where L is the length of the beating region while Δβ is the

difference between the propagation constants of the two
modes, which can be represented as22

i 2y
expjjj− zzz
λ(U2∞ 2‐U1∞ 2)
k V{
Δβ =
4πncladding ρ2 (3)

where Um∞ is the asymptotic value of the modes (U1∞

corresponds to HE11 = 2.405 and U∞ 2 corresponds to HE12 =
5.520),21 ρ is the radius of the tapered region, and V is the
normalized frequency given as
2πρ 2 2
V= (ncladding ‐ nmedium )1/2
λ (4)

where λ is the light wavelength and ncladding and nmedium are the
refractive indexes of the fiber cladding and the surrounding
medium of the tapered region, respectively. Based on the above
equations, it can be predicted that the output spectrum of the
optical microfiber is similar to a sinusoidal form as a function
of wavelength. From eqs 2−4, the spectrum shifts due to the Figure 2. (A−D) Simulated intensity mode profiles for HE11 and
change of the refractive index at the cladding−medium HE12 modes at the cross section of the tapered waist region, (E)
interface can be derived as23
i 2 2 y
enlarged area of the mode intensity profile at the cladding−air

Δλ = λjjjexp ( ‐ )‐1zzz
interface, (F) simulated light power of HE11 and HE12 modes at the

k V′ V {
tapered region with different down-taper lengths, Z, (G) simulated
(5) effective refractive index for HE11 (red: primary x and y-axes) and
HE12 (blue: primary x and y-axes) modes at different refractive
where V′ is the revised normalized frequency indexes of the cladding−medium interface and the phase difference
between the two modes (green: primary x-axis and secondary y-axis),
2πρ 2
V′ = (ncladding ‐(nmedium + Δnmedium)2 )1/2 and (H) spectrum shift as a function of the refractive index of the
λ + Δλ (6) cladding−medium interface; simulated and experimental results.

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ACS Appl. Mater. Interfaces 2019, 11, 28546−28553
ACS Applied Materials & Interfaces
Figure 3. (A) TEM image of the as-synthesized CDs, (B) FT-IR spectrum, (C) surface elemental mapping, and (D) EDS analysis of the CD-
functionalized optical microfiber. The inset shows the SEM image of the microfiber, and the area highlighted with a red dashed box is the measured
area.
intensity mode profiles for HE11 and HE12 modes at the cross
section of the 7.9 μm tapered waist region. Figure 2E shows
the enlarged R1 and R2 regions that are marked as dashed
boxes in Figures 2C,D, respectively. It was found that the
evanescent field of the HE12 mode has a larger amplitude than
the HE11 mode, indicating that the HE12 mode has a larger
penetration depth than the HE11 mode. The penetration depth
of the HE12 mode is calculated by determining the 1/e value of
its amplitude and was found to be approximately 540 nm,
which suffice for ferric ion sensing. On the other hand, Figure
2F shows the light power of HE11 and HE12 modes at the
tapered region with different down-taper lengths, Z. It can be
seen that the HE12 mode does not exist for an untapered fiber
(Z = 0 μm) but exists when Z > 0, where the fiber tapering was
initiated. The HE12 mode is being excited when the fiber radius
decreases abruptly, which justifies the stronger HE12 mode at a
shorter Z length. As the Z length increases, it can be
understood that the fiber radius decreases gradually; hence,
the HE12 mode is generally weaker owing to lesser energy
coupling from the HE11 mode. Based on the simulated result,
strongest energy coupling was found at Z = 1840 μm, where
the power intensity of the HE11 mode was at its minimum
while the power of the HE12 mode was at its maximum.
Therefore, the ideal down-taper length of the optical
microfiber was 1840 μm, which will result in the strongest
evanescent field for the interaction with the surrounding
medium. However, due to the limitation of our tapering
machine as well as to ensure the ease of handling, the down-
taper length was fixed at 3000 μm. Furthermore, Figure 2G
depicts the effective refractive indices for HE11 and HE12
modes across different refractive index values at the cladding−
medium interface of the tapered region and the corresponding
phase difference between the two modes. As described in eqs
1−5, the phase difference will contribute to an output
spectrum shift, as shown in Figure 2H. The simulated result
(red line) clearly shows an increase in the refractive index from
1.33 to 1.37 at the cladding−medium interface, leading to a red
shift of the spectrum. More importantly, it was found that a
refractive index difference of 0.002 resulted in a spectrum shift
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Research Article
of 5 nm, indicating the theoretical capability of our optical
microfiber to detect a minute change of the refractive index at
the fiber cladding−medium interface. We further validated
these simulation results with the fabricated microfiber sensor
using glycerin solutions of different refractive indices. The
experimental results (blue dot in Figure 2H) are found to be in
good agreement with the simulation results.
It has been reported that binding of heavy metal ions onto
the chelator-modified optical fiber sensor will cause refractive
index change at the cladding (metal-bonded on the chelator-
modified surface)−medium interface, which can be detected
by the resonance wavelength shift in the output signal.24,25
Likewise, we foresee that the binding of Fe3+ onto the surface
of the CD-functionalized optical microfiber (Figure 1C) will
induce a change in the refractive index at the cladding−
medium interface, which in turn modulates the phase
difference between the modes propagating along the tapered
length and eventually translates to wavelength shifts of the
interference spectrum. Therefore, the amount of detectable
Fe3+ can be quantified by correlating the output spectrum shift.
3.2. Characterization of CDs and CD-Functionalized
Optical Microfiber. The CDs were prepared using a one-step
microwave-assisted pyrolysis process, and the resulting CDs
were found to have an average size of 3.27 ± 0.58 nm, as
observed in Figure 3A. The surface properties of as-synthesized
CDs were analyzed using Fourier transform infrared spectros-
copy (FT-IR) and is shown in Figure 3B. A strong and broad
large peak centered at 3290 cm−1 is associated with the band of
O−H and N−H stretching, while the peaks found at around
2360 cm−1 are contributed by the S−H moiety.15 These peaks
correspond to the surface functional groups of the as-
synthesized CDs. On the lower wavenumber, absorption
peaks associated with CO and CC moieties were seen
at 1637 and 1514 cm−1, respectively. The presence of CDs on
the surface of the optical microfiber was validated using FT-IR,
and similar peaks were found on the CD-functionalized
microfiber, confirming successful functionalization of CDs on
the surface of the tapered region. In addition to this, Figure 3C
also shows the scanning electron microscopy (SEM) image of
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Figure 4. (A) Time vs sensor’s response at different concentrations of Fe3+ solutions, (B) calibration curve for the CD-functionalized optical
microfiber sensor and (inset) linear sensing response to changes in the Fe3+ concentration (n = 5), (C) sensor calibration curve fitted to the
Langmuir adsorption isotherm model (n = 5), and (D) CD-functionalized optical microfiber sensor response to various kinds of heavy metal ions
(n = 5).

Table 1. Previous Reported CD-Based Sensors for Fe3+ Sensing

sensing scheme linear detection range LOD refs
fluorescence quenching 0−0.7 μM 4.2 nM 30
fluorescence quenching 0−100 μM 0.96 μM 31
fluorescence quenching 0−16 μM 242 μM 32
fluorescence quenching 0−30 μM 21 nM 33
fluorescence quenching 0.3−546 μM 90 nM 34
fluorescence quenching 0−30 μM 15 nM 35
fluorescence quenching 1−78 μM 0.54 μM 36
fluorescence quenching 0.05−30 μM 13.5 nM 37
refractive index change using CDs-functionalized optical microfiber 0−5.37 μM (0−300 μg/L) 13.8 nM (0.77 μg/L) this work

the surface of a CD-functionalized microfiber, which has been
used for Fe3+ detection. Elemental analysis using energy-
dispersive X-ray spectroscopy (EDS) (Figure 3D) validated the
formation of Fe3+ chelates on the tapered region upon binding
to the as-synthesized CDs.
3.3. Quantification of Fe3+ in Aqueous Medium and
the Detection Limit. Figure 4A presents the responses of the
sensor to three different concentrations of Fe3+ solutions. A
negligible spectrum shift was observed when deionized water
was introduced into the sensor, indicating that no reaction has
occurred. Meanwhile, when 50 and 200 μg/L Fe3+ solutions
were introduced into the sensor, incremental spectrum shifts
were observed as a result of the interaction between CDs and
Fe3+ ions, which changed the refractive index at the surface
spectrum shift was found to linearly increase with the
increasing Fe3+ concentration. The functional groups of the
as-synthesized CDs serve as the electron donors and form
coordinate bonds with Fe3+ ions, which lead to the formation
of Fe3+ chelates on the surface of the tapered region.26 For this
reason, the refractive index at the tapered region-ambient
interface was altered, resulting in a shift in the output
spectrum. The sensor exhibits a linear detection range from 0
to 300 μg/L and sensitivity of 0.0061 nm/(μg/L).
The relationship between the measured output spectrum
shift and the amount of Fe3+ present was further analyzed
based on the Langmuir isotherm model to calculate the sensor
detection limit. The nonlinear expression of the Langmuir
isotherm model can be illustrated as follows27
i LC yz

interface. In both cases, the measured output spectrum shift
reached a plateau after 4 min. Hence, for the subsequent
measurements, the reaction time between the sensor and the
sample solution was fixed at 4 min.
The detection capability of the CD-functionalized optical
microfiber sensor was examined with Fe3+ solutions of
concentrations ranging from 0 to 3000 μg/L prepared using
deionized water. As shown in Figure 4B, the measured
28550
Δλ = Δλmax jjj zz
k 1 + LC {
(8)
where Δλ is the output spectrum shift as a result of Fe3+ ion
adsorption onto the CD-functionalized optical microfiber,
Δλmax is the maximum output spectrum shift induced by the
formation of a complete monolayer, C is Fe3+ concentration in
the aqueous solution, and L is the Langmuir constant. Using
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the nonlinear curve fitting based on eq 8, Figure 4C shows a sensor for on-site use. Figure S2 depicts the actual image of the
good correlation of the experimental data (R2 = 0.9687), and in-house portable interrogation system and the step-by-step
the Langmuir isotherm parameter values are found to be Δλmax procedures to perform Fe3+ detection using the CD-function-
= 2.4790 nm and K = 0.0052 (μg/L)−1. Omitting the effects of alized optical microfiber sensor. The inset of Figure S2A
inherent thermal and environmental noise originating from the portrays a CD-functionalized optical microfiber sensor
OSA, the LOD of the proposed sensor is primarily restricted packaged in a cartridge terminated with LC-type connectors
by the OSA resolution.28 To calculate the LOD, the Langmuir and mounted on the interrogation system. A round opening
isotherm equation can be rewritten as29 was made below the sensor window to introduce liquid
samples to the sensor as well as to minimize perturbation to
Δλ the microfiber during the insertion and removal of liquid
C LOD =
K(Δλmax ‐ Δλ) (9) samples. The details of the interrogation system have been
previously described in detail.14 Briefly, the linear calibration
Using the K and Δλmax determined previously and by curve equation in Figure 3B was programmed into the
substituting Δλ with 0.01 nm (resolution of the OSA), the interrogation system to correlate the output spectrum shift
calculated CLOD of the sensor was found to be 0.77 μg/L. In to the concentration of the Fe3+ present. The interrogation
summary, the developed sensor exhibited a lower LOD than system would then show the detected concentration values on
that of recently reported CD-based sensors listed in Table 1 the display screen. To perform the analysis of the real samples,
and the World Health Organization requirement (300 μg/L) the CD-functionalized optical microfiber sensor was immersed
of Fe3+ in drinking water. for twice in deionized water to ensure that the output spectrum
3.4. Effects of pH and Temperature. The stability of the was fixed, followed by immersing in the sample and left
CD-functionalized microfiber sensor was assessed by subject- undisturbed for 4 min to allow the reaction to proceed to
ing it to elevated temperatures at 30, 40, and 50 °C for 30 min. completion take place. Finally, the measured Fe3+ concen-
The annealed microfiber sensors were then used to measure tration can be read from the display screen.
aqueous samples containing 100 μg/L Fe3+. As shown in To evaluate the practicability of the CD-functionalized
Figure S1A (Supporting Information), a negligible temperature optical microfiber sensor for the environmental and biological
influence was observed, indicating the stability of the surface applications, tap water, phosphate-buffered saline (PBS),
coating. In addition, the sensitivity of the CD-functionalized Dulbecco’s modified eagle’s medium (DMEM), fetal bovine
microfiber sensor under different pH values ranging from pH 4
serum (FBS), and horse serum samples were employed to
to pH 9 was also investigated. From Figure S1B, the effect of
conduct the test. FBS and horse serum samples were subjected
pH variation was negligible from pH 4 to pH 7. However, as
to a 100-fold dilution before analysis. The iron ion content in
the pH increases above pH 7, the sensitivity of the sensor
each control sample was identified using inductively coupled
reduces significantly. This can be attributed to the weak
solubility of Fe3+ due to the formation of ferric hydroxide in mass spectroscopy (ICPMS) or based on formulation
alkaline medium.38 Therefore, the optimized medium pH for standards and are listed in Table S1. Furthermore, the samples
optimal sensing performance of the fabricated CD-function- were spiked with various concentrations of the standard Fe3+
alized microfiber sensor is investigated as weakly acidic to solution. As shown in Table 2, the test results obtained using
neutral, validating its potential applications in biosensing. the presented sensor were in accordance with the known
3.5. Selectivity Test of CD-Functionalized Microfiber
Sensor. To evaluate the selectivity of the CD-functionalized Table 2. Determination of Fe3+ in Environmental Samples,
optical microfiber sensor, the sensor was examined with Biological Buffers, and Biological Serums Using the CD-
various metal ion solutions including Al3+, Ca2+, Cd2+, Cs2+, Functionalized Optical Microfiber Sensor (n = 5)
Cu2+, Fe2+, Mg2+, Mn2+, Ni2+, Pb2+, and Zn2+, each at a
concentration of 100 μg/L at room temperature. Figure 4D spiked Fe3+
sample content measured concentration using
illustrates the measured concentration for each cation. type (μg/L) CDs-functionalized microfiber sensor (μg/L)
Negligible interference from other cations and exceptionally tap water 100 92.4 ± 6.9
high selectivity toward Fe3+ ions were observed. This result is tap water 200 204.3 ± 6.8
in accordance with the literature, which described the existence DMEM 0 96.1 ± 6.4
of hydroxyl, amine, and carboxylic groups on the surface of the DMEM 100 206.7 ± 13.0
CDs, which can specifically coordinate with Fe3+ ions.15 In PBS 100 110.6 ± 11.5
addition to this, a mixed-metal solution containing the above- PBS 200 207.0 ± 7.9
mentioned interfering ions was tested, with and without Fe3+ fetal 0 31.3 ± 2.6
to investigate the antijamming capability of the sensor. bovine
Notably, the sensor showed high affinity to Fe3+ despite the serum
presence of interfering metal ions. Slight interference was fetal 100 121.8 ± 12.0
bovine
observed with the mixture solution containing no Fe3+, which serum
could be attributed to the nonspecific binding of the interfering fetal 200 221.5 ± 10.1
ions. bovine
serum
3.6. Integration of CD-Functionalized Optical Micro-
horse 0 18.3 ± 2.6
fiber with Portable Interrogation System for the serum
Detection of Fe3+ in Tap Water and Biological Samples. horse 100 112.5 ± 11.1
In our previous work, we have developed an in-house portable serum
interrogation system, which was aimed at substituting the OSA horse 200 209.1 ± 12.5
and promoting the usability of the optical microfiber-based serum

28551 DOI: 10.1021/acsami.9b08934

ACS Appl. Mater. Interfaces 2019, 11, 28546−28553
ACS Applied Materials & Interfaces
spiked Fe3+ content. However, it is worth noting that for
DMEM samples, the Fe3+ concentrations detected were 2
times more than the spiked concentration. This is attributed to
the Sigma-Aldrich’s DMEM formulation that contains a trace
level of Fe3+, ∼100 μg/L. Likewise, 30 and 23 μg/L iron ion
content was found in FBS and horse serum samples (100-fold
dilution), respectively, after being tested using inductively
coupled mass spectroscopy. This explains the slight increment
of the measured concentration using the proposed sensor
compared to the spiked Fe3+ concentration. Overall, good
analytical agreement was achieved, further confirming the
reliability and practicability of the CD-functionalized optical
microfiber sensor for Fe3+ detection in environmental and
biological samples.
4. CONCLUSIONS
Nitrogen- and sulfur-codoped CDs were successfully prepared
in a one-step microwave-assisted pyrolysis method and
functionalized onto the tapered surface of an optical microfiber
as a chelator to enable metal ion detection at the solid phase.
The end product was used as a label-free Fe3+ sensor probe,
and it possesses excellent detection sensitivity, achieving an
LOD of 0.77 μg/L, calculated using the Langmuir isotherm
model and a linear detection range of 0−300 μg/L. More
significantly, the established sensor exhibited high selectivity to
Fe3+ compared to other types of metal ions. The presented
CD-functionalized optical microfiber sensor and the in-house
portable interrogation system have demonstrated the practic-
ability of the system to be used as a fast screening tool for on-
site detection, particularly for environmental monitoring and
biological sensing applications. Apart from the fact that this
work has introduced a new avenue of the CD-based sensing
mechanism, it is envisaged that the presented work can be
further extended to adopt other functional CDs or novel
nanomaterials as the sensing element owing to the versatility of
chemical modification on the surface of the developed optical
microfiber. Overall, this work has demonstrated the feasibility
of nanomaterial-based sensors to operate at the solid phase via
the integration with optical microfiber sensor technology.
■
ASSOCIATED CONTENT
*
S Supporting Information
The Supporting Information is available free of charge on the
ACS Publications website at DOI: 10.1021/acsami.9b08934.
Temperature and pH effects; procedures to perform
Fe3+ detection using the in-house portable interrogation
system; ICPMS results for control tap water and
biological samples (PDF)
■
AUTHOR INFORMATION
Corresponding Author
*E-mail: ktyong@ntu.edu.sg.
ORCID
Stephanie Hui Kit Yap: 0000-0002-2847-9936
Kok Ken Chan: 0000-0002-0592-4427
Ken-Tye Yong: 0000-0001-7936-2941
Author Contributions
† (17) Qiu, H.; Gao, S.; Chen, P.; Li, Z.; Liu, X.; Zhang, C.; Xu, Y.;
S.H.K.Y. and K.K.C. contributed equally to this work.
Notes
The authors declare no competing financial interest.
28552
Research Article
■ ACKNOWLEDGMENTS
This work was supported by the NRF-ANR Joint Call 2017
project (Grant no. M4197007.640) and MOE Tier 2 project
(MOE2017-T2-2-002; Grant no. M4020385.110)
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