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Fluorescent-Nitrogen-Doped Carbon Quantum Dots Derived from

Citrus Lemon Juice: Green Synthesis, Mercury(II) Ion Sensing, and
Live Cell Imaging
Aschalew Tadesse,* Mebrahtu Hagos, Dharmasoth RamaDevi, Kaloth Basavaiah, and Neway Belachew
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ABSTRACT: In this study, we report a green and economical hydrothermal

synthesis of fluorescent-nitrogen-doped carbon quantum dots (NCQDs) using
citrus lemon as a carbon source. The prepared NCQDs possess high water
Downloaded via 94.231.218.20 on February 21, 2020 at 18:06:53 (UTC).

solubility, high ionic stability, resistance to photobleaching, and bright blue

color under ultraviolet radiation with a high quantum yield (∼31%). High-
resolution transmission electron microscopy (HRTEM) results show that the
prepared NCQDs have a narrow size distribution (1−6 nm) with an average
particle size of 3 nm. The mercury ion (Hg2+) sensing efficiency of the NCQDs
was studied, and the result indicated that the material has high sensitivity, high
precision, and good selectivity for Hg2+. The limit of detection (LOD) is 5.3
nM and the limit of quantification (LOQ) is 18.3 nM at a 99% confidence
level. The cytotoxicity was evaluated using MCF7 cells, and the cell viabilities
were determined to be greater than 88% upon the addition of NCQDs over a
wide concentration range from 0 to 2 mg/mL. Based on the low cytotoxicity, good biocompatibility, and other revealed interesting
merits, we also applied the prepared NCQDs as an effective fluorescent probe for multicolor live cell imaging.

■ INTRODUCTION
At present, water contamination by heavy metals has become a
serious problem.1 Heavy metals are toxic to aquatic living
organisms and harmful to humans through food-chain
accumulation.2 Mercury is a heavy metal well known for its
extremely high toxicity and a threat to human life and the
whole environment.3 Inorganic mercury in water mainly
appears in the toxic form of the Hg2+ oxidation state. As per
the World Health Organization (WHO) recommendation, the
maximum uptake of Hg2+ is 0.3 mg/week and the maximum
acceptable limit in drinking water is 1 μg/L.4 The maximum
level of mercury in drinking water permitted by the US
Environmental Protection Agency is 2 ppb.5 In this regard,
monitoring the concentration of Hg2+ in water is very essential.
Techniques for accurate quantification of trace concentrations
of mercury such as atomic absorption spectroscopy6 and
inductively coupled plasma atomic emission spectrometry7
provide limits of detection up to parts per billion levels.
However, their excellent performance is achieved at a high cost
and time-consuming sample preparation and preconcentration
procedures. As an alternative, fluorescent detection methods
that can encompass a variety of advantages, including real-time
and remote measurements, sensitivity and selectivity against
competing analytes, and reliable, compact, and low-cost
detection, have been developed for sensing of Hg2+ ions in
an aquatic environment. Ncube et al.8 reported a fluorescent
sensing probe based on a naphthyl azo dye modified dibenzo-
18-crown-6-ether for the detection and determination of Hg2+
ions in water, and the probe showed high sensitivity and
selectivity for Hg2+ ions among various alkali, alkaline earth,
and transition-metal ions. Xia and Zhu9 used surface-modified
CdTe quantum dots as fluorescent probes in the sensing of
Hg2+ ions. However, the highly toxic effect of inorganic
quantum dots, low quantum yield, and easy photobleaching of
organic dyes have limited the application of these materials in
the real-environment detection of Hg2+ ions. Recently, Hg2+
ion probe based on the fluorescence quenching of carbon
quantum dots (CQDs) has been developed and received
intense attention because of its simplicity, sensitivity, and
selectivity.10−12
Carbon dots, which are also known as carbon quantum dots
(CQDs), are a class of fluorescent nanomaterials with ultralow
particle size (<10 nm). CQDs have interesting properties such
as unique bright and tunable photoluminescence, high water
solubility, excellent biocompatibility, and stability to photo-
degradation and photobleaching.13 In comparison with other
quantum dots that are derived from heavy-metal precursors,
CQDs are environmentally green and nontoxic to cell.14
Received: September 27, 2019
Accepted: February 12, 2020

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Because of these important properties, the CQDs have
attracted immense attention in various applications such as
analytical sensing,15 bioimaging,14 and photocatalysis.16
Much progress has been made in the synthesis of
photoluminescent CQDs using different methods. Researchers
frequently used the hydrothermal method since it is a facile,
eco-friendly, one-step process with a low synthesis cost as
compared to other methods.17 Carbonization carried out by
the hydrothermal method at low temperature with self-created
pressure is a green method because no pollutant is released
into the environment. Different natural products like orange
juice18 and amino acids19 have been used as precursors to
prepare fluorescent CQDs. Investigating new and green
precursors for the economical synthesis of fluorescent CQDs
is important. In the search for new precursors for carbon
quantum dots, using environmentally green plant materials is
becoming most encouraging. However, the quantum yield of
most of the carbon quantum dots prepared from plant
materials is relatively low.20−24 In the search for high quantum
yield carbon quantum dots from natural products, we
investigated the synthesis of NPQDs from lemon fruit juice
as a source of carbon.
Studies have shown that citric acid is an excellent precursor
for the synthesis of CQDs because it has well-known low
carbonization temperature,18 and amine-containing molecules
such as ethylenediamine are able to passivate CQDs with
amines and form nitrogen-doped carbon quantum dots, thus
increasing the quantum yield and selectively sensing analytes,
such as Hg2+, Cu2+, and Cr6+ ions.14 In spite of the controversy
regarding the mechanism of the photoluminescence (PL) of
doped CQDs, it is agreed that trap states with different energy
levels can be introduced by surface groups, which result in
excitation-dependent fluorescence.25
The sour taste of lemon juice is due to the presence of a high
concentration of citric acid.26 Motivated by these, we
synthesized water-soluble highly fluorescent-nitrogen-doped
carbon quantum dots (NCQDs) with high quantum yield (QY
∼ 31%) using eco-friendly green hydrothermal synthesis
methods utilizing lemon juice as a precursor and ethylenedi-
amine as the co-reagent. The fluorescence intensity of the
prepared NCQDs is based on the pH value, and they have high
photostability under different salt conditions, high photo-
bleaching resistance, and high water solubility. The cell
viability study using the 3-(4,5-dimethylthiazol-2-yl)-2,5-
diphenyl tetrazolium bromide (MTT) assay of the MCF7
cell line indicated that the as-synthesized NCQDs are
nontoxic. The prepared NCQDs were used for Hg2+ ion
sensing in water and cell imaging, and the results revealed that
they have great potential in sensing and biological application.
■ EXPERIMENTAL SECTION
Chemicals and Reagents. Citrus lemons were collected
fresh from the nearby market in Andhra University, India, and
NCQDs were prepared from the juice of these lemons using
the hydrothermal method. All the reagents were of analytical
grade and used without any further purification. Milli-Q water
was used throughout the analysis.
Synthesis of NCQDs. Twenty milliliters of filtered fresh
lemon juice and 2 mL of ethylenediamine co-reagent were
mixed in 100 mL hydrothermal autoclave and kept in a furnace
at 200 °C for 3 h. The as-obtained black paste was cooled
down to room temperature and then dissolved in 15 mL of
water and centrifuged at 3000 rpm for 15 min to separate
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insoluble matter. Then, dichloromethane was added to the
brown solution of NCQDs and centrifuged at 3000 rpm for 20
min to remove unreacted organic moieties. The separated
upper aqueous layer was then centrifuged at 12 000 rpm for 20
min thrice to remove larger-sized particles, and a brown-
yellowish supernatant was finally obtained. NCQDs with
smaller particle sizes were separated using column chromatog-
raphy taking silica gel as the stationary phase and dichloro-
methane as the solvent. The as-prepared NCQDs were stored
at 4 °C for characterization and used for applications.
Characterization. High-resolution transmission electron
microscopy (HRTEM, Jeol/JEM 2100, LaB6) operated at 200
kV was used to determine the morphological and micro-
structural properties of the prepared NCQDs. Further
morphological and compositional information of the samples
was determined using field emission scanning electron
microscopy (FESEM, Zeiss Ultra-60) equipped with an X-
ray energy-dispersive spectroscope. PANalytical X’pert pro X-
ray diffraction (XRD) using Cu Kα1 radiation (45 kV, 1.54056
Å; scan rate of 0.02°/s) was used to determine the crystalline
pattern. The IR spectra were obtained over the range of 400−
4000 cm−1 using an Fourier transform infrared (FTIR)
spectrometer (Bruker). The emission properties of the
NCQDs were determined with a Fluoromax-4 spectrofluor-
ometer (HORIBA Scientific). The absorbance spectra of
NCQDs were recorded using a UNICAM UV 500
spectrophotometer (Thermo Electron Corporation).
Quantum Yield. The following eq 1 was used to calculate
the quantum yield (QY) by taking quinine sulfate in 0.1 M
H2SO4 (the quantum yield of 54% at 365 nm) as a reference.27
The QY of NCQDs was determined by following the reported
protocol. The UV−vis absorption at 365 nm and PL emission
spectra (excited at 365 nm) of carbon dots and reference was
measured, respectively. To reduce the reabsorption effects, the
absorbance was kept under 0.05 at the excitation wavelength.
The QY can be calculated using the following equation
QY samp = QY ref (Isamp/Iref )(A ref /A samp)(nsamp 2 /nref 2) (1)
where QYsamp and QYref are the quantum yields; Isamp and Iref
are emission intensities; Asamp and Aref are the UV−vis
absorbances; and nsamp and nref are refractive indexes of the
sample and reference, respectively.
Sensing of Hg2+. The fluorescent (FL) assay of Hg2+ was
carried out at room temperature using fluorescence quenching
of 25 μg/mL of NCQDs in phosphate-buffered solution (PBS)
at pH 7.4 (absorbance of solution = 0.05). For the detection of
Hg2+ ion, the NCQDs solution was taken in a cuvette and 30
μL of different concentrations of Hg2+ was added, and finally,
the volume was adjusted to 3 mL by the NCQDs solution. The
FL emission spectra at 365 nm excitation were measured after
reaction for 5 min at room temperature. Selectivity study was
done using competing ions in place of Hg2+ and a mixture of
other cations with Hg2+. The reproducibility of the method
was investigated by recording the FL spectra of nine solutions
prepared side by side containing 1 × 10−8 M Hg2+ prepared in
the same procedure as mentioned above.
Cytotoxicity Test. The biocompatibility of NCQDs was
tested on human breast adenocarcinoma (MCF7) cells using
the MTT assay. Cytotoxicity was evaluated by determining the
activity of mitochondrial enzymes in live MCF7 cells to
transform the yellow MTT solution to an insoluble purple
formazan. MCF7 cells were cultured in 96-well tissue culture
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Scheme 1. Scheme Showing Reaction of Precursors to Produce NCQDs
plates at a density of 1 × 104 cells per well and then the cells
were incubated in a medium containing different doses of
NCQDs for 24 h. Subsequently, the medium was removed and
the cells were washed with the phosphate-buffered solution
(PBS). Fresh medium with 10 μL of 0.5 mg/mL MTT solution
was supplemented to each well and incubated for 4 h and then
the medium was replaced with 150 μL of dimethyl sulfoxide
(DMSO) followed by shaking for 15 min to dissolve the
formazan crystals. The absorbance of each well was measured
at 570 nm using a multimode microplate reader (Biotek,
Cytation3). The untreated cells were used as controls for
determining the cell viability from the following equation
% cell viability = (A 570 in treated sample
/A 570 in untreated sample) × 100% (2)
Cell Imaging. The bioimaging potential of NCQDs was
investigated using MCF7 cells. The MCF7 cells were cultured
in 6-well culture plates at a density of 105 cells per well in
Dulbecco’s modified eagle medium (DMEM) having 10% fetal
bovine serum (FBS) and incubated for 24 h at 37 °C and 5%
CO2. Then, the medium was replaced with a fresh medium of
0.025 mg/mL NCQDs and incubated for 6 h. After that the
cells were washed thrice with PBS to remove extracellular
NCQDs, fixed with 4% paraformaldehyde and mounted using
50% glycerol. Images were obtained using “Zeiss LSM 510
Meta confocal Microscopy” at laser excitations of 405, 488, and
561 nm.
■ RESULTS AND DISCUSSION
Characterization. Upon hydrothermal treatment of
precursors at 200 °C for 3 h and after separation as per
procedures, the brown solution of NCQDs was obtained, as
shown in Scheme 1. Figure 1 shows the absorption and
emission spectra of the as-prepared NCQDs. The prepared
NCQDs show two typical absorption peaks at 245 and 353
nm, as shown in Figure 1 (black solid line), which extended
with a tail to the visible region. The absorption peak at 245 nm
could be assigned to the π−π* transition of aromatic −CC−
bonds in the sp2-hybridized domain of graphitic core and the
other peak at 353 nm could be assigned to the n−π* transition
of −CO, C−N, or −C−OH bonds in the sp3-hybridized
domains, which may be from hydroxyl (−COOH) or amine
(−NH2) group on the surface of NCQDs.28−31 The aqueous
solution of NCQDs with brown-yellowish color appears bright
blue under ultraviolet radiation (inset in Figure 1b), which
indicates the bright luminescence of the prepared NCQDs.
Figure 1 (blue broken line) indicates the emission spectra of
the blue luminescent NCQDs, with excitation at 360 nm and
emission at 452 nm. Full width at half-maximum (FWHM)
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Figure 1. UV−vis absorption (solid black) and fluorescence spectra
(blue broken) of (a) NCQDs in daylight and (b) NCQDs under
ultraviolet radiation.
value obtained at this particular fluorescence emission is 75
nm, indicating the narrow size distribution of the synthesized
NCQDs.32−35
To further study the optical properties of the as-prepared
NCQDs, a fluorescence study was carried out at different
excitation wavelengths. To clarify the concept of photo-
luminescence, the 2D FL emission spectra of the NCQDs can
be divided into two parts: one with excitation wavelengths
from 300 to 410 nm (Figure 2a) and the other with the
excitation wavelength from 410 to 490 nm (Figure 2b). As
seen from the figures, the sample shows fluorescence with
symmetrical peaks. The fluorescence emission intensity
increases as the excitation wavelength increases from 310 to
360 nm, and the maximum emission intensity is obtained at
the excitation wavelength of 360 nm. Further increase in
excitation wavelengths results in a decrease in emission
intensity. Overall, as the excitation wavelengths increase, the
emission wavelengths shift to longer wavelengths (red shift).
Figure 2c depicts the fluorescence emission−excitation spectra
of the NCQDs, with excitation wavelengths increasing from
300 to 400 nm in 4 nm steps and showing symmetrical
emission peaks. Figure 2d indicates the excitation and emission
contour map of NCQDs showing emission at different
excitation wavelengths. The fluorescence of NCQDS may be
due to the π-plasmon absorption in the core carbon
nanoparticles,36 and, to large extent, photoluminescence is
highly affected by the surface chemistry of NCQDs.37 The as-
synthesized NCQDs show a high FL quantum yield (QY ∼
31%) relative to the standard quinine sulfate. This relatively
high QY is due to nitrogen doping in NCQDs, as heteroatom
doping may significantly improve the QY of carbon dots and N
doping may provide the most effective enhancement to
QY.38−40 As shown in Table 1, compared to the NCQDs
synthesized using different plant materials as precursor, the
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Figure 2. (a) FL emission spectra of NCQDs at excitation wavelengths from 300 to 410 nm, (b) FL emission spectra of NCQDs at excitation
wavelengths from 410 to 490 nm, (c) excitation−emission fluorescence spectra of NCQDs (excitation from 300 to 400 nm and corresponding
emission), and (d) excitation and emission contour map of NCQDs.

Table 1. Comparison of Fluorescence Quantum Yields of NCQDs synthesized from citrus lemon juice in this work has a
CQDs Prepared Using Different Plants high QY.41−46 NCQDs showed the conversion FL in addition
to strong down-converted FL properties. Figure S3a shows the
sl. no. plant materials as precursor quantum yield (%) refs
FL spectra as NCQDs are excited with a longer-wavelength
1 Syzygium cumini fruits 5.9 21
light with emissions centered at a wavelength of 450 nm, which
2 Chionanthus retusus fruit 9 22
is comparable to that of the down-converted FL peaks.
3 fennel seeds 9.5 23
The ionic strength of the as-synthesized NCQDs was
4 quince fruit 8.55 41
investigated at different concentrations (0.25−2 M NaCl), and
5 lotus root 19.0 43
6 ginkgo fruit 3.33 44
it was obtained that the change in the concentration of salt has
7 carrot juice 5.16 45
no significant effect on the fluorescence of NCQDs (Figure
8 unripe Prunus mume juice 16 47 3a). In addition, NCQDs have shown high FL stability in that
9 Citrus aurantium juice 19.9 48 neither significant shift in emission wavelength nor significant
10 citrus lemon juice 31 present study reduction in FL intensity is observed upon continuous
exposure to ultraviolet radiation from 1 to 45 min (Figure

Figure 3. (a) Fluorescence intensity of NCQDs at excitation wavelength of 365 nm indicating the ionic strength of NaCl solution of concentration
0.25−2 M. (b) Stability after irradiation at different times and after preservation for 1 and 3 months. (c) pH effect in acidic media with a pH of 6−
2. (d) pH effect in basic media with pH of 8−12.

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Figure 4. (a, b) Representative TEM and HRTEM (inset) images. (c) SAED of NCQDs and (d) particle size distribution histogram.
3b) and after 3 months of storage. The effect of pH on the
fluorescence property of NCQDs was investigated and the
fluorescence intensity of the NCQDs depends on the pH of
the solution but there is no shift in emission wavelength.
Decreasing pH from 6 to 2 increased the FL intensity (Figure
3c) and increasing the pH from 8 to 12 decreased the FL
intensity (Figure 3d). These results described that under low
pH values, the NCQDs, probably, exist as isolated species in
the aqueous solution; the nanoparticles agglomerated with
increasing pH value because of interactions such as hydrogen
bonds between the carboxyl groups.46 The change in
fluorescence intensity with pH change should result from the
transition between isolated C−NH2 in acidic conditions and
agglomerated CNH in alkaline conditions. As shown in
Figure S8, the surface nitrogen of NCQDs-OH existed in the
form of imino groups in the alkaline environment. However,
the imino groups could be transformed into amino groups in
acidic conditions.
The morphological properties of the as-synthesized NCQDs
were confirmed by HRTEM (Figure 4a,b). The results
indicated that the NCQDs are uniformly distributed quasi-
spherical nanoparticles with narrow size distribution. Particle
sizes of the NCQDs are of diameter in the range of 1−6 nm
with an average of 3 nm based on a statistical analysis of more
than 90 dots (histogram in Figure 4d). Figure 4c shows the
selected area electron diffraction (SAED) of the NCQDs, the
holes indicating the particle formation and only two bright
spots observed indicating the amorphous nature. As shown in
the inset of Figure 4a, the as-synthesized NCQDs exhibit a
lattice spacing of 0.19 nm, which is similar to the (100) facet of
the sp2 graphitic carbon.49 X-ray diffraction patterns show a
broad and intense diffraction peak centered at 2θ = 23° and a
weak peak at 2θ = 42°, which is assigned to the (002) and
(101) diffraction patterns of graphitic carbon, as shown in
Figure 7a, which indicates the amorphous nature of the
NCQDs and is in accordance with previous structure analysis
on disordered amorphous graphitic CQDs.50
Information regarding the surface functional group of
NCQDs was investigated by Fourier transform infrared
spectroscopy (FTIR). As can be seen in the spectrum (Figure
6b), there is a broad overlapping strong bands in the range of
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3070−3600 cm−1, which can be due to the stretching vibration
of the O−H and N−H bonds. The vibrational band of the C−
O bond at 1112 cm−1 confirms the presence of primary −OH
groups on the surface of NCQDs. The appearance of bands at
2934 cm−1 and in the range of 1040−1300 cm−1 illustrates the
existence of aliphatic C−H and C−N/C−O functional groups,
respectively. The sharp band at 1546 cm−1 can be assigned to
the N−H vibration and deformation band, which indicates the
presence of an amino functional group. In addition, the
existence of characteristic stretching and bending vibrational
bands for C−H and stretching vibrations for CC in aromatic
hydrocarbons at 3060, 770, and 1434 cm−1 confirms the
presence of the aromatic skeleton (sp2-hybridized carbon) in
the prepared NCQDs. Hence, the FTIR analysis shows the
presence of functional groups such as alcohols, amines,
aromatic rings, and carbonyls on the surface and sp2/sp3
core carbon in NCQDs derived from citrus lemon juice. The
presence of different functional groups on the surface makes
the NCQDs highly water-soluble.51−53
Raman spectrometer analysis also confirmed the structural
properties of NCQDs. As shown in Figure 5, the Raman
spectrum of the NCQDs exhibits two peaks at 1348 and 1547
cm−1, corresponding to the D and G bands of graphitic carbon,
respectively. The ratio of ID/IG is 1.03, which is characteristic
of the disorder amorphous nature and the ratio of sp3/sp2
carbon, implying structural defects in the synthesized NCQDs.
Figure 5. Raman spectra of NCQDs.
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Figure 6. (a) XRD peak of NCQDs. (b) FTIR spectrum. (c) Energy dispersive spectroscopy (EDS) spectrum (inset elemental composition of
NCQDs and FESEM image).
FESEM images of a paper sheetlike layer in Figure 6c inset
indicate the amorphous nature of NCQDs. The result from the
elemental composition analysis of the energy-dispersive X-ray
spectroscopy (EDX) spectrum (Figure 6c) reveals the
presence of C, O, and N in the as-synthesized material,
indicating the formation of nitrogen-doped carbon quantum
dots.
Quantitative Assay. The mechanism to determine Hg2+
using fluorescence quenching of NCQDs is based on the
interaction of oxygen/nitrogen-containing groups over the
NCQDs surface with Hg2+ cations. The oxygen/nitrogen-
containing groups have a good affinity to form a coordination
bond with Hg2+ ions by donating electron pair from the
functional terminal to the metal ions, resulting in the
fluorescence quenching effect.54 The fluorescence quenching
of NCQDs by Hg2+ ions presumably due to facilitating
nonradiative electron/hole recombination/annihilation
through partial electron transfer process in coordinate bond
formation.55,56 The fluorescence sensing performance of the
nitrogen-doped carbon quantum dots based on fluorescent
quenching was evaluated by adding different concentrations of
Hg2+ (0−100 μM) into the NCQD solution. As shown in
Scheme 2, the fluorescence of NCQDs quenched in the
presence of Hg2+ ion and the fluorescence quenching intensity
depends on the concentration of Hg2+. The fluorescence
spectra in the presence of various concentrations of Hg2+ in
Scheme 2. Scheme Showing Fluorescence Quenching of
NCQDs by Hg2+ Ions
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NCQDs are shown in Figure 7a. With increasing concentration
of Hg2+, the fluorescence intensity of the NCQDs was
gradually decreased.57 A linear relationship between the
fluorescence quenching factor of NCQDs and the concen-
tration of Hg2+ was obtained within a linear range of 0.001−1
μM with a correlation coefficient R2 of 0.995, as shown in
Figure 7b.
The linear regression equation53 is
(I0 − I )/I0 = 0.1616[Hg 2 +] + 0.2418 (3)
where I0 and I are the fluorescence intensities of NCQDs
solutions at 465 nm without and in the presence of Hg2+ ions,
respectively, and [Hg2+] is the concentration of Hg2+ ions.
Detection limit determined based on the blank is 5.3 nM (t. σ,
n = 9, where σ is the standard deviation and t is taken from a
one-sided Student’s t distribution for n − 1 degrees of freedom
at a 99% confidence level) and precision is 1.32% relative
standard deviation (RSD; 0.01 μM, n = 11). The limit of
quantification for this method is 18.3 nM (10σ, n = 9) at a 99%
confidence level.
Selectivity. To study the selectivity of Hg2+ ions over other
competing cations, 30 μL of 1 × 10−4 M solution of cations
(Na+, K+, Mg2+, Mn2+, Cu2+, Ni2+, Ca2+, Co2+, Cd2+, Zn2+, Fe2+,
Fe3+, Sn2+, Ba2+, Hg2+, Pb2+, Al3+) were added to NCQD
solution in phosphate-buffered solution of pH 7.4 and makeup
to 3 mL with final concentration of 1 μM. Fluorescence
spectrum was recorded after 5 min of addition and compared
with the fluorescence intensity of blank (3 mL of NCQDs in
phosphate-buffered solution). As shown in Figure 7c, the
addition of Hg2+ and Fe3+ ions noticeably reduces the intensity
of NCQDs, whereas the effect of other cations is negligible. To
further investigate the interference of the above cations with
Hg2+ ions, 25 μL of 1 × 10−4 M solution of cations (Na+, K+,
Mg2+, Mn2+, Cu2+, Ni2+, Ca2+, Co2+, Cd2+, Zn2+, Fe2+, Fe3+,
Sn2+, Ba2+, Pb2+, Al3+) and 30 μL of 1 × 10−4 M of Hg2+ were
taken in a cuvette containing NCQDs and the volume adjusted
to 3 mL and then the spectrum was recorded. Figure 7d
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Figure 7. (a) Fluorescence spectra of NCQDs solution in the presence of different concentrations of Hg2+ and (b) corresponding calibration curve.
(c) Fluorescence quenching response of NCQDs to different metal ions with the same concentration (1 μM). (d) Specific FL quenching response
of NCQDs to Hg2+ ions in the presence of other cations.

illustrates that the presence of cations (Na+, K+, Mg2+, Mn2+,

Cu2+, Ni2+, Ca2+, Co2+, Cd2+, Zn2+, Fe2+, Sn2+, Ba2+, Pb2+, Al3+)
with Hg2+ has no effect on the fluorescence sensing of Hg2+
because the fluorescence decrease is the same as that for the
solution containing only Hg2+. However, the presence of Fe3+
causes more decrease, although it is insignificant. Hence,
NCQDs can be used for selective fluorescence sensing of Hg2+
ions in aqueous solution.
Real Water Analysis. To evaluate the feasibility of the
fluorescent NCQDs for Hg2+ sensing, different concentrations
of Hg2+ added in water samples were determined by the spike
recovery method, and the results are listed in Table 2. It can be

Table 2. Result of Hg2+ Detection in Real Water Samples Figure 8. NCQDs Cell Viability Using MTT Assay.
Using Fluorescent NCQDs
revealed the low toxicity and excellent biocompatibility of the
concentration of Hg2+(μM) prepared NCQDs derived from lemon juice, which made them
amount amount % %RSD a suitable fluorescent probe for bioimaging application.44
sample added found recovery (n = 3) Fluorescence imaging potential of NCQDs was tested by
tap water 1 0.250 0.256 100.8 2.5 treating MCF7 cells with 0.025 mg/mL of NCQDs derived
tap water 2 0.040 0.041 95.2 4.3 from citrus lemon juice. Cell images were recorded with
packed water 1 0.100 0.098 96.0 2.8 confocal microscopy at laser excitations of 405, 488, and 561
packed water 2 0.100 0.097 95.0 3.7 nm. Figure 9 shows the confocal microscopic images of MCF7
cells after incubation with NCQDs. The obtained bright-field
seen that the recoveries of the samples were found to be in the image after incubation indicates the viability of the cells
range of 95−101% and the RSDs were less than 5.0%. The (Figure 9a). After excitation at 405, 488, and 561 nm, bright
results showed that it was an applicable method for the blue, green, and red luminescence in the cytoplasm region was
determination of Hg2+ in water samples. observed, respectively, indicating the excellent cell membrane
Cytotoxicity Test and Live Cell Imaging. The feasibility permeability of NCQDs. 58 The results indicate that
of NCQDs derived from lemon juice for potential bioimaging fluorescent, water-soluble NCQDS derived from citrus lemon
juice can be used as an excellent fluorescent bioimaging probe.

applications was evaluated by the cytotoxicity test through the
MTT assay using human breast adenocarcinoma (MCF7)
cells. Results of cytotoxicity studies performed using various
concentrations of NCQDs (0.312−2 mg/mL) are shown in
Figure 8. The NCQDs exhibited low cytotoxicity, with cells
maintaining the viability of about 88% for MCF7 cells at a high
concentration of 2 mg/mL after 24 h incubation. The results
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■
CONCLUSIONS
A facile, environmentally green hydrothermal method was used
to synthesize highly photofluorescent, water-soluble nitrogen-
doped carbon quantum dots (NCQDs) using abundantly
available fruit, citrus lemon juice as a carbon source and
ACS Omega XXXX, XXX, XXX−XXX
ACS Omega http://pubs.acs.org/journal/acsodf
Figure 9. (a−d) Fluorescence images of NCQDs labeled MCF7 cells under bright field, 405, 488, and 561 nm excitations, respectively.
ethylenediamine as a coreagent. The obtained NCQDs exhibit
high quantum yield, narrow size distribution, highest
fluorescence intensity, good water solubility, excellent stability,
and good resistance to ionic strength and photobleaching.
From an environmental perspective, investigation of the
sensing efficiency of NCQDs for Hg2+ ions in water samples
shows that the material is suitable to develop selective sensors
with high accuracy (LOD = 5.3 nM, LOQ = 18.3 nM) and
high precision. From the MTT assay, we observed that the
prepared NCQDs show low cytotoxicity and excellent
biocompatibility. The multicolor living cell imaging of the
NCQDs was investigated, and we conclude that the prepared
NCQDs may be used as an eco-friendly fluorescent nanoma-
terial for potential in vitro or in vivo multicolor imaging and
sensing applications.
■
*
ASSOCIATED CONTENT
sı Supporting Information
The Supporting Information is available free of charge at
https://pubs.acs.org/doi/10.1021/acsomega.9b03175.
Optimization of synthesis method, quantum yield of
synthesized NCQDs, FESEM images of NCQDs,
upconverted fluorescence of NCQDS, electronic proper-
ties of NCQDs, 1H NMR spectra and 13C NMR of
NCQDs, quenching effect of Hg2+ on fluorescence of
NCQDs, pH effect on fluorescence of NCQDs, and
carbon quantum dots dFL quenching based methods for
Hg2+ detection (PDF)
■
AUTHOR INFORMATION
Corresponding Author
Aschalew Tadesse − Department of Applied Chemistry, Adama
Science and Technology University, Oromia 1888, Ethiopia;
orcid.org/0000-0001-6962-1479; Phone: +251-
0901732147; Email: asche.best@gmail.com
H https://dx.doi.org/10.1021/acsomega.9b03175
Article
Authors
Mebrahtu Hagos − Faculty of Natural and Computational
Sciences, Woldia University, Woldia 400, Ethiopia
Dharmasoth RamaDevi − AU College of Pharmaceutical
Sciences, Andhra University, Visakhapatnam 530003, India
Kaloth Basavaiah − Department of Inorganic and Analytical
chemistry, Andhra University, Visakhapatnam 530003, India
Neway Belachew − Department of Chemistry, Debrebirahan
University, Debre-Berhan 445, Ethiopia
Complete contact information is available at:
https://pubs.acs.org/10.1021/acsomega.9b03175
Notes
The authors declare no competing financial interest.
■
ACKNOWLEDGMENTS
The authors are grateful for the support from the Ministry of
Education, Federal Democratic Republic of Ethiopia and
Adama Science and Technology University. Thanks also go to
Dr. P. Shyamala (Head, Department of Physical, Nuclear
Chemistry and Chemical Oceanography, Andhra University)
for allowing us to use their instruments.
■
REFERENCES
(1) Asrari, E. Heavy Metal Contamination of Water and Soil: Analysis,
Assessment, and Remediation Strategies; CRC Press, 2014.
(2) Olsson, P.-E.; Kling, P.; Hogstrand, C. Mechanisms of Heavy
Metal Accumulation and Toxicity in Fish. Metal Metab. Aquat.
Environ. 1998, 6, 321−350.
(3) Gopalakrishnan, S.; Thilagam, H.; Raja, P. V. Comparison of
Heavy Metal Toxicity in Life Stages (spermiotoxicity, Egg Toxicity,
Embryotoxicity and Larval Toxicity) of Hydroides elegans. Chemo-
sphere 2008, 71, 515−528.
(4) World Health Organization. Guidelines for Drinking-Water
Quality; World Health Organization, 2006.
(5) Wu, D.; Huang, W.; Duan, C.; Lin, Z.; Meng, Q. Highly
Sensitive Fluorescent Probe for Selective Detection of Hg2 + in DMF
Aqueous Media. Inorg. Chem. 2007, 46, 1538−1540.
ACS Omega XXXX, XXX, XXX−XXX
ACS Omega http://pubs.acs.org/journal/acsodf
(6) Hatch, W. R.; Ott, W. L. Determination of Submicrogram
Quantities of Mercury by Atomic Absorption Spectrophotometry.
Anal. Chem. 1968, 40, 2085−2087.
(7) Han, F. X.; Dean Patterson, W.; Xia, Y.; Maruthi Sridhar, B. B.;
Su, Y. Rapid Determination of Mercury in Plant and Soil Samples
Using Inductively Coupled Plasma Atomic Emission Spectroscopy, a
Comparative Study. Water, Air, Soil Pollut. 2006, 170, 161−171.
(8) Ncube, P.; Krause, R. W. M.; Ndinteh, D. T.; Mamba, B. B.
Fluorescent Sensing and Determination of Mercury (II) Ions in
Water. Water SA. 2014, 40, No. 175.
(9) Xia, Y.-S.; Zhu, C.-Q. Use of Surface-Modified CdTe Quantum
Dots as Fluorescent Probes in Sensing Mercury (II). Talanta 2008,
75, 215−221.
(10) Wu, H.; Tong, C. Nitrogen- and Sulfur-Codoped Carbon Dots
for Highly Selective and Sensitive Fluorescent Detection of Hg2 Ions
and Sulfide in Environmental Water Samples. J. Agric. Food Chem.
2019, 67, 2794−2800.
(11) Zhang, Y.; He, Y. H.; Cui, P. P.; Feng, X. T.; Chen, L.; Yang, Y.
Z.; Liu, X. G. Water-Soluble, Nitrogen-Doped Fluorescent Carbon
Dots for Highly Sensitive and Selective Detection of Hg2 in Aqueous
Solution. RSC Adv. 2015, 5, 40393−40401.
(12) Liang, J. Y.; Han, L.; Liu, S. G.; Ju, Y. J.; Li, N. B.; Luo, H. Q.
Carbon Dots-Based Fluorescent Turn Off/on Sensor for Highly
Selective and Sensitive Detection of Hg2 and Biothiols. Spectrochim.
Acta, Part A 2019, 222, No. 117260.
(13) Song, Y.; Zhu, S.; Yang, B. Bioimaging Based on Fluorescent
Carbon Dots. RSC Adv. 2014, 4, No. 27184.
(14) Wang, J.; Zhang, P.; Huang, C.; Liu, G.; Leung, K. C.-F.; Wáng,
Y. X. J. High Performance Photoluminescent Carbon Dots for In
Vitro and In Vivo Bioimaging: Effect of Nitrogen Doping Ratios.
Langmuir 2015, 31, 8063−8073.
(15) Qian, Z. S.; Chai, L. J.; Huang, Y. Y.; Tang, C.; Jia Shen, J.;
Chen, J. R.; Feng, H. A Real-Time Fluorescent Assay for the
Detection of Alkaline Phosphatase Activity Based on Carbon
Quantum Dots. Biosens. Bioelectron. 2015, 68, 675−680.
(16) Wang, H.; Zhuang, J.; Velado, D.; Wei, Z.; Matsui, H.; Zhou, S.
Near-Infrared- and Visible-Light-Enhanced Metal-Free Catalytic
Degradation of Organic Pollutants over Carbon-Dot-Based Carboca-
talysts Synthesized from Biomass. ACS Appl. Mater. Interfaces 2015, 7,
27703−27712.
(17) Hua, J.; Yang, J.; Zhu, Y.; Zhao, C.; Yang, Y. Highly Fluorescent
Carbon Quantum Dots as Nanoprobes for Sensitive and Selective
Determination of Mercury (II) in Surface Waters. Spectrochim. Acta,
Part A 2017, 187, 149−155.
(18) Sahu, S.; Behera, B.; Maiti, T. K.; Mohapatra, S. Simple One-
Step Synthesis of Highly Luminescent Carbon Dots from Orange
Juice: Application as Excellent Bio-Imaging Agents. Chem. Commun.
2012, 48, 8835−8837.
(19) Pei, S.; Zhang, J.; Gao, M.; Wu, D.; Yang, Y.; Liu, R. A Facile
Hydrothermal Approach towards Photoluminescent Carbon Dots
from Amino Acids. J. Colloid Interface Sci. 2015, 439, 129−133.
(20) Feng, X.; Zhang, Y. A Simple and Green Synthesis of Carbon
Quantum Dots from Coke for White Light-Emitting Devices. RSC
Adv. 2019, 9, 33789−33793.
(21) Bhamore, J. R.; Jha, S.; Singhal, R. K.; Kailasa, S. K. Synthesis of
Water Dispersible Fluorescent Carbon Nanocrystals from Syzygium
cumini Fruits for the Detection of Fe Ion in Water and Biological
Samples and Imaging of Fusarium avenaceum Cells. J. Fluoresc. 2017,
27, 125−134.
(22) Atchudan, R.; Nesakumar Jebakumar, T.; Chakradhar, D.;
Perumal, S.; Shim, J.-J.; Lee, Y. R. Facile Green Synthesis of Nitrogen-
Doped Carbon Dots Using Chionanthus retusus Fruit Extract and
Investigation of Their Suitability for Metal Ion Sensing and Biological
Applications. Sens. Actuators, B 2017, 246, 497−509.
(23) Dager, A.; Uchida, T.; Maekawa, T.; Tachibana, M. Synthesis
and Characterization of Mono-Disperse Carbon Quantum Dots from
Fennel Seeds: Photoluminescence Analysis Using Machine Learning.
Sci. Rep. 2019, 9, No. 14004.
I https://dx.doi.org/10.1021/acsomega.9b03175
Article
(24) Meng, W.; Bai, X.; Wang, B.; Liu, Z.; Lu, S.; Yang, B. Biomass-
Derived Carbon Dots and Their Applications. Energy Environ. Mater.
2019, 2, 172−192.
(25) Chatzimitakos, T.; Stalikas, C. Recent Advances in Carbon
Dots. J. Carbon Res. 2019, 5, No. 41.
(26) Penniston, K. L.; Nakada, S. Y.; Holmes, R. P.; Assimos, D. G.
Quantitative Assessment of Citric Acid in Lemon Juice, Lime Juice,
and Commercially-Available Fruit Juice Products. J. Endourol. 2008,
22, 567−570.
(27) Magde, D.; Rojas, G. E.; Seybold, P. G. Solvent Dependence of
the Fluorescence Lifetimes of Xanthene Dyes. Photochem. Photobiol.
1999, 70, No. 737.
(28) Bourlinos, A. B.; Stassinopoulos, A.; Anglos, D.; Zboril, R.;
Karakassides, M.; Giannelis, E. P. Surface Functionalized Carbogenic
Quantum Dots. Small 2008, 4, 455−458.
(29) Dimos, K. Carbon Quantum Dots: Surface Passivation and
Functionalization. Curr. Org. Chem. 2016, 20, 682−695.
(30) Vieira, K. O.; Bettini, J.; de Oliveira, L. F. C.; Ferrari, J. L.;
Schiavon, M. A. Synthesis of Multicolor Photoluminescent Carbon
Quantum Dots Functionalized with Hydrocarbons of Different Chain
Lengths. Carbon 2017, 124, No. 729.
(31) Huang, Y.; Liao, W. Hierarchical Carbon Material of N-Doped
Carbon Quantum Dots in-Situ Formed on N-Doped Carbon
Nanotube for Efficient Oxygen Reduction. Appl. Surf. Sci. 2019,
495, No. 143597.
(32) Perez-Guzman, M. A.; Ortega-Amaya, R.; Ortega Lopez, M. In
Advances on Synthesis of Highly Luminescent Carbon Quantum Dots by
Citric Acid Carbonization, 2017 14th International Conference on
Electrical Engineering, Computing Science and Automatic Control
(CCE). IEEE: Mexico City, Mexico, 2017.
(33) Kumar, V. B.; Sheinberger, J.; Porat, Z.; Shav-Tal, Y.;
Gedanken, A. A Hydrothermal Reaction of an Aqueous Solution of
BSA Yields Highly Fluorescent N Doped C-Dots Used for Imaging of
Live Mammalian Cells. J. Mater. Chem. B 2016, 4, 2913−2920.
(34) Ren, G.; Zhang, Q.; Li, S.; Fu, S.; Chai, F.; Wang, C.; Qu, F.
One Pot Synthesis of Highly Fluorescent N Doped C-Dots and Used
as Fluorescent Probe Detection for Hg2 and Ag in Aqueous Solution.
Sens. Actuators, B 2017, 243, 244−253.
(35) Atchudan, R.; Nesakumar Jebakumar, T.; Aseer, K. R.; Perumal,
S.; Karthik, N.; Lee, Y. R. Highly Fluorescent Nitrogen-Doped
Carbon Dots Derived from Phyllanthus acidus Utilized as a
Fluorescent Probe for Label-Free Selective Detection of Fe3 Ions,
Live Cell Imaging and Fluorescent Ink. Biosens. Bioelectron. 2018, 99,
303−311.
(36) Fernando, K. A.; Sahu, S.; Liu, Y.; Lewis, W. K.; Guliants, E. A.;
Jafariyan, A.; Wang, P.; Bunker, C. E.; Sun, Y.-P. Carbon Quantum
Dots and Applications in Photocatalytic Energy Conversion. ACS
Appl. Mater. Interfaces 2015, 7, 8363−8376.
(37) Baker, S. N.; Baker, G. A. Luminescent Carbon Nanodots:
Emergent Nanolights. Angew. Chem., Int. Ed. 2010, 49, 6726−6744.
(38) Shan, X.; Chai, L.; Ma, J.; Qian, Z.; Chen, J.; Feng, H. B-Doped
Carbon Quantum Dots as a Sensitive Fluorescence Probe for
Hydrogen Peroxide and Glucose Detection. Analyst 2014, 139,
2322−2325.
(39) Wang, Y.; Kim, S.-H.; Feng, L. Highly Luminescent N, S- Co-
Doped Carbon Dots and Their Direct Use as mercury(II) Sensor.
Anal. Chim. Acta. 2015, 890, 134−142.
(40) Gu, D.; Hong, L.; Zhang, L.; Liu, H.; Shang, S. Nitrogen and
Sulfur Co-Doped Highly Luminescent Carbon Dots for Sensitive
Detection of Cd (II) Ions and Living Cell Imaging Applications. J.
Photochem. Photobiol., B 2018, 186, 144−151.
(41) Ramezani, Z.; Qorbanpour, M.; Rahbar, N. Green Synthesis of
Carbon Quantum Dots Using Quince Fruit (Cydonia oblonga)
Powder as Carbon Precursor: Application in Cell Imaging and As3
Determination. Colloids Surf., A 2018, 549, 58−66.
(42) Zhang, W.; Jia, L.; Guo, X.; Yang, R.; Zhang, Y.; Zhao, Z. Green
Synthesis of up- and down-Conversion Photoluminescent Carbon
Dots from Coffee Beans for Fe3 Detection and Cell Imaging. Analyst
2019, 144, No. 7421.
ACS Omega XXXX, XXX, XXX−XXX
ACS Omega http://pubs.acs.org/journal/acsodf Article

(43) Gu, D.; Shang, S.; Yu, Q.; Shen, J. Green Synthesis of Nitrogen-
Doped Carbon Dots from Lotus Root for Hg(II) Ions Detection and
Cell Imaging. Appl. Surf. Sci. 2016, 390, 38−42.
(44) Li, L.; Li, L.; Chen, C.-P.; Cui, F. Green Synthesis of Nitrogen-
Doped Carbon Dots from Ginkgo Fruits and the Application in Cell
Imaging. Inorg. Chem. Commun. 2017, 86, 227−231.
(45) Liu, Y.; Liu, Y.; Park, M.; Park, S.-J.; Zhang, Y.; Akanda, M. R.;
Park, B.-Y.; Kim, H. Y. Green Synthesis of Fluorescent Carbon Dots
from Carrot Juice for in Vitro Cellular Imaging. Carbon Lett. 2017, 21,
61−67.
(46) Carvalho, J.; Santos, L. R.; Germino, J. C.; Terezo, A. J.;
Moreto, J. A.; Quites, F. J.; Freitas, R. G. Hydrothermal Synthesis to
Water-Stable Luminescent Carbon Dots from Acerola Fruit for
Photoluminescent Composites Preparation and Its Application as
Sensors. Mater. Res. 2019, 22, No. 12736.
(47) Atchudan, R.; Nesakumar Jebakumar, T.; Sethuraman, M. G.;
Lee, Y. R. Efficient Synthesis of Highly Fluorescent Nitrogen-Doped
Carbon Dots for Cell Imaging Using Unripe Fruit Extract of Prunus
Mume. Appl. Surf. Sci. 2016, 384, 432−441.
(48) Fatahi, Z.; Esfandiari, N.; Ehtesabi, H.; Bagheri, Z.; Tavana, H.;
Ranjbar, Z.; Latifi, H. Physicochemical and Cytotoxicity Analysis of
Green Synthesis Carbon Dots for Cell Imaging. EXCLI J. 2019, 18,
454−466.
(49) Dong, Y.; Pang, H.; Yang, H. B.; Guo, C.; Shao, J.; Chi, Y.; Li,
C. M.; Yu, T. Carbon-Based Dots Co-Doped with Nitrogen and
Sulfur for High Quantum Yield and Excitation-Independent Emission.
Angew. Chem. 2013, 125, 7954−7958.
(50) Hao, A.; Guo, X.; Wu, Q.; Sun, Y.; Cong, C.; Liu, W. Exploring
the Interactions between Polyethyleneimine Modified Fluorescent
Carbon Dots and Bovine Serum Albumin by Spectroscopic Methods.
J. Lumin. 2016, 170, 90−96.
(51) Yan, Z.; Shu, J.; Yu, Y.; Zhang, Z.; Liu, Z.; Chen, J. Preparation
of Carbon Quantum Dots Based on Starch and Their Spectral
Properties. Luminescence 2015, 30, 388−392.
(52) Liu, X.; Zheng, J.; Yang, Y.; Chen, Y.; Liu, X. Preparation of N-
Doped Carbon Dots Based on Starch and Their Application in White
LED. Opt. Mater. 2018, 86, 530−536.
(53) Wang, Z.-X.; Ding, S.-N. One-Pot Green Synthesis of High
Quantum Yield Oxygen-Doped, Nitrogen-Rich, Photoluminescent
Polymer Carbon Nanoribbons as an Effective Fluorescent Sensing
Platform for Sensitive and Selective Detection of silver(I) and
mercury(II) Ions. Anal. Chem. 2014, 86, 7436−7445.
(54) Abdullah Issa, M.; Z Abidin, Z.; Sobri, S.; Rashid, S.; Adzir
Mahdi, M.; Azowa Ibrahim, N.; Y Pudza, M. Facile Synthesis of
Nitrogen-Doped Carbon Dots from Lignocellulosic Waste. Nanoma-
terials 2019, 9, No. 1500.
(55) Jeong, C. J.; Roy, A. K.; Kim, S. H.; Lee, J.-E.; Jeong, J. H.; In,
I.; Park, S. Y. Fluorescent Carbon Nanoparticles Derived from Natural
Materials of Mango Fruit for Bio-Imaging Probes. Nanoscale 2014, 6,
15196−15202.
(56) Mewada, A.; Pandey, S.; Shinde, S.; Mishra, N.; Oza, G.;
Thakur, M.; Sharon, M.; Sharon, M. Green Synthesis of
Biocompatible Carbon Dots Using Aqueous Extract of Trapa
bispinosa Peel. Mater. Sci. Eng., C 2013, 33, 2914−2917.
(57) Wang, C.; Sun, D.; Chen, Y.; Zhuo, K. A Hydrothermal Route
for Synthesizing Highly Luminescent Sulfur- and Nitrogen-Co-Doped
Carbon Dots as Nanosensors for Hg2. RSC Adv. 2016, 6, 86436−
86442.
(58) Sachdev, A.; Gopinath, P. Green Synthesis of Multifunctional
Carbon Dots from Coriander Leaves and Their Potential Application
as Antioxidants, Sensors and Bioimaging Agents. Analyst 2015, 140,
4260−4269.

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