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Cite This: ACS Omega 2019, 4, 1581−1591 http://pubs.acs.org/journal/acsodf

Antibacterial Nitrogen-doped Carbon Dots as a Reversible

“Fluorescent Nanoswitch” and Fluorescent Ink
Satyesh Raj Anand, Anshu Bhati, Deepika Saini, Gunture, Neetu Chauhan, Prateek Khare,
and Sumit Kumar Sonkar*
Department of Chemistry, Malaviya National Institute of Technology, Jaipur, Jaipur 302017, India

ABSTRACT: The present finding describes an efficient facile approach for the
fabrication of nitrogen-doped carbon dots (N-CDs) as a “fluorescent nanoswitch”.
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Highly fluorescent blue-light-emitting N-CDs have been synthesized via a simpler

hydrothermal method using 2,2′-(ethylenedioxy)-bis(ethylamine) and malic acid as
the precursors. N-CDs showed excitation-dependent and pH-independent emission
along with a quantum yield of ∼25%. The blue fluorescent emission of N-CDs has
Downloaded via 171.247.77.90 on September 12, 2021 at 10:03:00 (UTC).

been selectively “turned off” (quenching of fluorescence (FL)) during the sensing of
Cr(VI) with 0.02 μM limit of detection and further been selectively “turned on”
(restoration of FL) on sensing ascorbic acid, compared with other metal cations and
biomolecules tested. For testing the practical applicability of N-CDs, the switchable
reversibility of the fluorescent nanoswitch has been tested for up to four cycles on
the basis of FL “on−off−on”. Furthermore, the toxicological test showed the
antibacterial effect of the N-CDs on the tested Gram-positive Staphylococcus aureus
and Gram-negative Escherichia coli cells. Additionally, these N-CDs can also be used
as a fluorescent ink for imaging purposes.

1. INTRODUCTION Co(II)35 especially because of their FL emission properties,

Fluorescent carbon dots (CDs)1−6 are zero-dimensional
carbon materials consisting of variable composition of sp2-/
sp3-hybridized carbon atoms with abundant functional groups
over the shell, due to which they exhibit unique fluorescence
(FL) properties.1 Since their discovery,2 CDs have triggered a
great interest in a wide range of applications, from
bioimaging,3,7 sensing,8,9 drug delivery,10,11 photocatalysis,12
energy storage,13 and fluorescent ink14,15 to disease diag-
nosis,16,17 and also in the field of agriculture18 because of their
superior unique properties such as the biocompatibility, robust
chemical inertness, nontoxicity,19 and stability.5,20−22 How-
ever, to accomplish more, presently the long-known method of
doping has been articulating the optical properties of CDs as in
the form of emergent doped-CDs.7,23,24 The doping of
heteroatoms significantly affects the physicochemical and
photochemical properties of CDs.7,24 Of all of the dopant
materials, the doping of N has been a bit more feasible
concerning its easier insertion in the carbonaceous matrix of
CDs because of its comparable size to that of carbon along
with the extra valence electron. Additionally, the emphasis on
escaping the complicated synthesis methodologies also has
been in demand, such as the exploration of newer sustainable
approaches based on the hydrothermal,25 solvothermal26,27
and microwave-assisted2,3,28 methodologies for the facile
fabrication of doped CDs as these are easier and scalable
methodologies with an extra advantage of ease in reproduci-
bility. From the many potential applications, currently, N-CDs
are being used for sensing the heavy metal ions like Cr(VI),29
Hg(II),9 Fe(III),30 Cu(II),31 Zn(II),32 Cd(II),33,34 and
“turn-off” and “turn-on”.
Cr(VI) is highly toxic in nature as compared to its lower
valent state Cr(III), causing several adverse impacts on the
environment as well as on the human health.36,37 Thus, their
discharge into the environment through industrial and other
anthropogenic activities needs to be monitored thoroughly.
Therefore, it is essential to continuously observe the level of
Cr(VI) in water media, which needs to be up to 0.9 μM,
possibly via using simpler techniques with accuracy even at its
lower level.29 In addition, the detection of ascorbic acid (AA)
(generally known as vitamin C) also has been important for
human health as AA is protecting us from many diseases.
Although a few reports have already been documented for the
detection of Cr(VI) and AA in the literature,38,39 the
development of simpler and economic methodologies has
always been in demand. For this, fluorescent properties of the
CDs2,3,40 and doped CDs like N-CDs have been explored on
the basis of their abilities of FL sensing.
The present finding depicts a simpler methodology for the
fabrication of N-CDs and their use as a reversible “fluorescent
nanoswitch” for selective sensing of Cr(VI) and AA. N-CDs
show excellent stability and solubility in aqueous medium, and
their fluorescent properties do not change on addition of
various other ions (interference study) and with the variation
in the pH of the solution, which may offer the practical
Received: November 15, 2018
Accepted: January 2, 2019
Published: January 17, 2019

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Scheme 1. Schematic Representation Showing the Synthesis and Multifunctional Application of N-CDs

Figure 1. (a) TEM micrograph of N-CDs; (b) particle size distribution histogram of N-CDs; (c) HRTEM image of N-CDs, and graphitic fringes
are marked with yellow circles; (d) HRTEM image showing a graphitic arrangement in a single N-CD.

application of N-CDs in the field of sensing as a reversible aqueous phase. Additionally, these were used as a blue-emitting
fluorescent nanoswitch. Also, these N-CDs worked as an
antibacterial agent for inhibiting the growth of the tested
Gram-positive S. aureus and Gram-negative E. coli cells in the fluorescent ink for imaging purposes.
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Figure 2. (a) UV−vis absorption spectrum of N-CDs (the inset shows the photographs comparing the aqueous solution of N-CDs under day light
(left) and UV light (right)); (b) FL emission spectra of N-CDs at varying excitations from 320 to 450 nm (the inset shows the stability of FL for 2
h under the continuous irradiation at 320 nm); (c) photoluminescence excitation spectra recorded at 445 nm emission; and (d) FL intensity of N-
CDs in different pH solutions.

Figure 3. (a) FT-IR and (b) survey XPS of N-CDs; (c) element composition and their corresponding high-resolution scans for: (d) C1s, (e) N1s,
and (f) O1s.

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Figure 4. (a) Photographic image of the aqueous solution of N-CDs with different metal ions under the UV-light illumination showing the selective
sensing of Cr(VI) (turn-off); (b) FL spectra of N-CDs on addition of different metal ions (λex: 320 nm); (c) FL intensity ratio (Io − I)/Io of N-CD
solution with different metal ions; (d) photographic image showing the selectivity of AA toward regaining the FL intensity of the maximally
quenched solution of the N-CD−Cr(VI) complex with different biomolecules; (e) FL spectra of the N-CD−Cr(VI) complex showing the regain in
the FL intensity on selective addition of AA (λex: 320 nm) with different metal ions under UV-light illumination; (f) FL intensity ratio (Io − I)/Io of
the N-CD−Cr(VI) complex with different biomolecules.

2. RESULTS AND DISCUSSION
A simpler hydrothermal technique has been used for the
synthesis of N-CDs by mixing 2,2′-(ethylenedioxy)-bis-
(ethylamine) and DL-malic acid. The schematic representation
depicting the hydrothermal synthesis and diverse applications
of N-CDs is shown in Scheme 1. The as-synthesized blue-light-
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emitting N-CDs show excellent solubility with excitation-
dependent emission having quantum yield (QY) ∼ 25%. These
N-CDs work as a reversible nanoswitch up to four cycles for
the sensing of Cr(VI) as “FL turn-off” and AA as “FL turn-on”.
The N-CDs also show antibacterial activity and work as a blue
fluorescent ink.
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2.1. Microscopic and Spectroscopic Analyses. The
morphological and microstructural analyses of N-CDs were
performed by transmission electron microscopy (TEM) and
high-resolution TEM (HRTEM). The TEM image of N-CDs
in Figure 1a shows the uniform distribution of N-CDs, along
with their size distribution histogram, Figure 1b, with Gaussian
fitting, which confirms that N-CDs own a size distribution
between 2 and 7 nm with an average diameter of ∼4.4 ± 0.87
nm. More structural information from Figure 1c confirms the
quasi-spherical nature of N-CDs (encircled by yellow circles).
The lattice spacing of N-CDs imaged by HRTEM (Figure 1d)
is 0.32 nm, which highlights the crystalline nature of N-CDs.
The optical properties of N-CDs (Figure 2) were
characterized by UV−vis and FL spectroscopy techniques.
N-CDs exhibit strong absorbance in the UV region, which
further extended throughout the visible region. The absorption
spectrum in Figure 2a displays three characteristic peaks: first
at ∼284 nm corresponding to π−π* transitions of the aromatic
sp2 domains and rest two peaks at ∼320 nm (a small hump)
and at ∼668 nm can be attributed to n−π* transitions due to
trapping of excited-state energy by the surface states, which is a
characteristic property of the graphene-like structures.41 The
N-CDs show a yellowish-brown color (Figure 2a; inset left)
under normal day light, which turns blue under the
illumination of UV light (Figure 2a; inset right). The emissive
properties of N-CDs were further studied at different excitation
wavelengths varying from 320 to 450 nm, as shown in Figure
2b. N-CDs feature excitation-wavelength-dependent emission
in the range of 420−520 nm, which is attributed to particle size
distribution with various lateral dimensions or emissive states
owing to different surface states, traps, and edge struc-
tures.42−45 The N-CDs show good photostability up to 2 h
with negligible bleaching under the continuous excitation of
320 nm, as shown in the inset of Figure 2b. The highest FL
emission intensity was observed at 445 nm (under excitation of
320 nm). The photoluminescence excitation study of N-CDs
shows three excitation peaks: at ∼389 nm (3.19 eV), ∼361 nm
(3.43 eV), and ∼252 nm (4.92 eV), as displayed in Figure 2c,
supporting the various emitting centers over the surface of N-
CDs. Along with this, the influence of the change in the pH on
the FL properties of N-CDs was investigated (Figure 2d, which
shows almost negligible effect of the pH on the FL intensity of
N-CDs).
N-Doping and the surface functionalization of the N-CDs
were further confirmed by Fourier transform infrared (FT-IR)
analysis. Figure 3a displays the FT-IR spectrum of N-CDs
showing the two significant peaks merged with small weak
vibrations located in the 3400−2800 cm−1 region. The broad
and merged peaks at around ∼3497 and ∼3295 cm−1
correspond to the −NH and −OH stretching vibrations,
respectively. A weak peak at ∼3080 cm−1 corresponds to the
(C−H) stretching vibration. The medium peaks at around
∼2920 and 2869 cm−1 arise due to the presence of (−C−H)
stretching vibrations. A strong peak at around ∼1644 cm−1 is
attributed to the presence of (CO/CN) stretching
vibrations46 of the amide group, whereas the peak at 1532
cm−1 is due to (OC−NH/CC) stretching vibrations.47
The weak peaks at ∼1454 and ∼1350 cm−1 correpond to the
(−C−N) stretching vibration and −C−NH stretching
vibration,48 respectively. The peak at ∼1287 cm−1 corresponds
to the epoxy group (C−O−C). A strong and sharp peak at
∼1090 cm−1 is attributed to the alkoxy (−C−O) stretching
vibration.49 The X-ray photoelectron spectroscopy (XPS)
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analysis was carried out to reveal the extent of N-doping and
for characterization of different binding species of carbon,
nitrogen, and oxygen in N-CDs. The XPS survey spectrum of
N-CDs, Figure 3b, shows peaks at ∼285.6, ∼400.5, and ∼533.0
eV corresponding to C1s (79.9%), N1s (8.1%), and O1s
(12.0%), respectively with their element composition as shown
in Figure 3c. Figure 3d shows the high-resolution C1s
spectrum, which can be deconvoluted into five peaks at
284.1 eV (CC), 284.6 eV (C−C), 285.5 Ev (C−N), 286.4
eV (C−O), and 288.0 eV (CO). Figure 3e shows the high-
resolution N1s spectrum, which depicts peaks at 399.3, 400.9,
and 401.2 eV associated with C−N−C, −N(C)3, and −N−H
binding, respectively. In the high resolution spectrum of O1s,
Figure 3f, three main components correspond to the C−O,
CO, and O−CO bindings located at 531.9, 532.8, and
534.1 eV, respectively. N-CDs exhibit negatively charged
surface functionalized groups supported by the FT-IR and XPS
analyses, which generate localized energy levels42 with different
emission centers. These emissive centers prefer nonradiative
recombination of charge carriers, present in the intrinsic states
with the utmost defects responsible for the emission which
leads to the characteristic excitation-dependent FL emission.
2.2. N-CDs as Fluorescent Nanoswitch for Sensing of
Cr(VI) and AA. The present work describes the newer
perspective of N-CDs as a FL-based reversible fluorescent
nanoswitch. For the fabrication of the reversible fluorescent
nanoswitch, the idea requires the initial turn-off in the FL
emission of the N-CDs via the addition of some quencher and
further regaining of the FL of the quenched solution after the
addition of some other specific molecule. For the same, herein,
the FL emission of N-CDs was selectively quenched (turn-off)
to its maximum via the addition of Cr(VI), followed by
regaining of FL emission of the quenched solution (turn-on)
via the addition of AA. N-CDs selectively sensed Cr(VI) and
showed strong quenching in their FL emission in comparison
to that from many other metal ions tested, such as Bi(III),
Co(II), Cu(II), Ni(II), Sr(III), Zn(II), Hg(II), and Cr(III)
ions. The selective, specific quenching affinity of Cr(VI) was
observed from the photographic images of the respective
solutions in the UV-light illumination, as shown in Figure 4a.
The selective FL quenching of N-CDs was observed in the case
of Cr(VI), whereas the other metal ions did not show
quenching.
In addition to photographic images, the selective FL
quenching of N-CDs by Cr(VI) was also demonstrated by
FL spectroscopy, as shown in Figure 4b. A qualitative change
in the FL intensity for the peak centered at ∼445 nm of N-CDs
decreased specifically with Cr(VI), compared with the other
metal ions. The selective sensing of Cr(VI) has been displayed
using (Io − I)/Io, where Io and I are the intensities of N-CDs
without and with metal ions, respectively, as shown in Figure
4c. However, the novelty, being presented here, was the
restoration of turn-on of the quenched emission of N-CDs via
the selective addition of AA. The basic idea was simple, that is,
just to release the loosely bound Cr(VI) from the surface of
the N-CDs, which would result in the regaining of the FL
intensity. Figure 4d shows the UV-illuminated photographic
images of the respective solution showing the specific
selectivity of AA toward the turn-on of the FL from the
maximally quenched solution of the N-CD−Cr(VI) complex.
From the many other common interfering biomolecules tested
such as adenine (Ad), L-cyteine (Cy), dextrose (Dt), L-ascorbic
acid (AA), guanine (Gu), L-proline (Pr), sulphonilic acid (SA),
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Figure 5. (a) Schematic illustration showing the plausible mechanism for the cycle of N-CDs as a reversible fluorescent nanoswitch for “on−off−
on” and (b) photographic image under UV-illumination for the four cycles, supporting the concept of N-CDs as a reversible fluorescent
nanoswitch.
thymine (Th), and glycine (Gl), the specific selectivity of AA
toward Cr(VI) can easily be explained by the long-known
favorable bonding interactions between AA and Cr(VI)
because of the higher affinity of AA toward Cr(VI) compared
with N-CDs.50,51 The quenched FL of the N-CD−Cr(VI)
complex turned on after subsequent addition of AA, Figure 4e.
The same, selective turn-on in the FL intensity of the
quenched solution of the N-CD−Cr(VI) complex using (Io −
I)/Io with different biomolecules has been displayed in Figure
4f.
Moreover, the reusability of this reversible fluorescent
nanoswitch concerning its cyclability toward the turn-off and
turn-on while selectively sensing Cr(VI) and AA was checked
and is demonstrated in Figure 5 up to four cycles. Figure 5a
demonstrates the possible mechanism for the working of the
reversible fluorescent nanoswitch on the basis of the long-
known DPC assay,3,52−54 selectively used for the detection of
Cr(VI). The quenching of the N-CDs FL can be explained by
ion-selective nature of surface functional groups of N-CDs and
superior charge/energy transfer effects.41 Based on the DPC
assay, while interacting with AA, almost all Cr(VI) gets
converted into Cr(III) and forms an AA−Cr complex.44,55 The
reversibility of the fluorescent nanoswitch can be easily
visualized under UV-light illumination, as displayed in Figure
5b. The interaction between N-CDs and Cr(VI) leads to a
loosely bound complex, that is, the N-CD−Cr(VI) complex,
that results in turn-off of its visible FL. Furthermore, this
loosely bound Cr(VI) may further have been attracted toward
AA because of the well-known strong affinity toward
Cr(VI).56−58
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As the negative consequences of the presence of toxic
Cr(VI) in water as a contaminant are well known, its active
sensing followed by its removal from water is important.
Owing to this, a FL-based study was carried out showing the
effect of varying concentrations of Cr(VI), in the range of
0.02−400 μM, on the FL intensity of N-CDs toward a
quantitative determination of Cr(VI). On gradually increasing
the concentration of Cr(VI), N-CD emission showed a linear
relation with quenching, as shown in Figure 6a. The zoomed
image in Figure 6b (of the area marked with a red rectangular
box) clearly shows a linear trend. The FL-quenching effect of
N-CDs can be quantitatively determined by the following
Stern−Volmer equation.
Io/I = 1 + KSV[Cr(VI)]
where Io is the FL intensity of N-CDs without Cr(VI), I is the
FL intensity in the presence of Cr(VI), and KSV is the static
Stern−Volmer constant. Figure 6c shows a plot of Io/I versus
increasing conc. of Cr(VI) (0.02−90 μM) for the Cr(VI)−N-
CDs system, which shows a linear relation with the regression
coefficient (R2). The KSV value of ∼0.0018 × 106 L mol−1 is
obtained from the slope of a linear fitted curve with a good
correlation coefficient (R2) of 0.9936. The limit of detection
was found to be ∼0.02 μM for Cr(VI), as calculated by taking
the FL intensity to be equal to ∼3 times the standard deviation
of the intensity (blank) divided by KSV. The synthesized N-
CDs can significantly determine the lower permissible limits of
Cr(VI), ∼0.9 μM, in drinking water as defined by the World
Health Organization.29 The same trend with the turn-on in the
FL intensity via the selective addition of AA has been shown in
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Figure 6. (a) FL quenching spectra of N-CDs by the addition of Cr(VI) at varying concentrations (0−400 μM); (b) zoomed FL spectra of (a); (c)
Stern−Volmer plot of the quenching of N-CDs emission intensity centered at 445 nm by Cr(VI); (d) FL spectra after the addition of AA in the
NCD−Cr(VI) complex (with concentration range 0−350 μM) in the N-CDs/Cr(VI)/AA system (FL turn-on); (e) zoomed FL spectra of (d)
(0−120 μM) and (f) Stern−Volmer plot of the FL turn-on of N-CDs with a linear fitting by the continuous addition of AA.

Figure 7. (a) Effect on the FL intensity in the presence of foreign metal ions with coexisting Cr(VI) and (b) corresponding tolerance limits for
sensing 5 mM Cr(VI) using N-CDs.

Figure 6d. Figure 6e shows the zoomed image (of the area
marked with a rectangular box) of the FL intensities with the
stepwise addition of AA showing the turn-on in the FL. A
linear relationship has been shown in Figure 6f for the
increasing concentration of AA verses Io/I of the FL intensity,
which turned on for the maximally quenched solution of the
N-CD−Cr(VI) complex. The limit of detection for AA was
observed to be ∼0.01 μM, as calculated from the slope of the
linear fitted curve (Figure 6f) with a good correlation
coefficient (R2) of 0.981.
2.3. Tolerance Study for Sensing Cr(VI). Toward the
practical applications of N-CDs as a reversible fluorescent
nanoswitch, their tolerance limit under the influence of several
coexisting metal ions has been analyzed. Figure 7a does not
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show any apparent changes in the selectivity for Cr(VI), even
in the presence of several coexisting ions. The tolerance limits
with different coexisting metal ions are presented in a tabular
form Figure 7b. The highly selective sensing of Cr(VI) up to 5
mM was revealed with negligible interference from foreign
metal ions (∼2-fold higher than that of Cr(VI)).
2.4. Antibacterial Effects of N-CDs Exposure to the E.
coli and S. aureus Bacterial Cells. The as-synthesized N-
CDs showed a strong antibacterial effect on the two different
strains of bacterial cells tested, that is, E. coli and S. aureus. The
antibacterial activity was tested by optical density (OD)
methods, as shown in Figure 8a,c, for E. coli and S. aureus,
respectively. The optical density of both types of bacteria
decreased on increasing the dose amount beyond 25 μL of N-
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Figure 8. Optical density (OD)-based growth bars at 600 nm (OD600) with the standard error showing the antibacterial effect of N-CDs at
different concentrations tested on (a) E. coli and (c) S. aureus cells. The same was analyzed based on the well-plate experiment for (b) E. coli and
(d) S. aureus.
Figure 9. Photographic image of paper-cut graphic of different animal stamps using N-CDs as a fluorescent ink under (a) day light and (b) UV
light illumination.
CDs per mL of bacterial media. In the case of E. coli, 100%
antibacterial activity was observed when the concentration was
25 μL of N-CDs per mL of media, whereas in the case of S.
aureus, a little strong strength of N-CDs was required but its
growth suppressed sharply at a higher amount of N-CDs (75
μL mL−1). The difference in antibacterial activity can be
attributed to the difference in the outer structure of E. coli,
Gram-negative bacteria, and S. aureus, Gram-positive bacteria,
and needs to be investigated in detail.
2.5. N-CDs as Fluorescent Ink. N-CDs can be used as a
fluorescent ink because of their excellent FL properties as
nowadays the fluorescent ink is being popularized for printing
purposes and in future can be used for the purpose of
information storage and encryption. Figure 9 shows the
pictorial image of different animal stamps using the N-CDs
imprinted on the butter paper under day light (as control
(Figure 9a)) and UV light irradiation (Figure 9b).
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3. CONCLUSIONS
N-CDs were used for the selective and sensitive FL sensing of
Cr(VI) and AA based on the simpler FL turn-off/turn-on
mechanism in aqueous medium. The sensing ability of N-CDs
as a reversible fluorescent nanoswitch allows comfortable
naked-eye-based detection, with high selectivity−sensitivity,
tolerance, and cycling stability under simpler operational
conditions. The antibacterial activity of N-CDs can be explored
in detail for the development of an antibacterial nanoprobe for
successful inhibition of bacterial growth and contamination.
Because of the excellent FL stability of the N-CDs, they have
the potential to be used as a fluorescent ink. This strategy,
together with the simplicity, can further expand the application
of N-CDs in the analytical field as a fluorescent nanoswitch for
sensing other systems along with inhibiting the growth of
bacterial cells.
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4. EXPERIMENTAL SECTION
4.1. Materials. All metal ions used for the selectivity test
were of nitrate salts, except for potassium dichromate
(K2Cr2O7), mercuric chloride (HgCl2), barium chloride
(BaCl2), calcium chloride (CaCl2), and all were purchased
from Sigma-Aldrich, India. Adenine (Ad), L-cyteine (Cy),
dextrose (Dt), L-ascorbic acid (AA), guanine (Gu), L-proline
(Pr), sulphonilic acid (SA), thymine (Th), and glycine (Gl)
were also purchased from Sigma-Aldrich, India. All of the
reagents were of analytical grade and used as received. The
deionized (DI) water was used throughout the synthesis and
sensing experiments unless otherwise stated.
4.2. Synthesis of N-CDs. N-CDs were synthesized
through a hydrothermal reaction using 2,2′-(ethylenedioxy)-
bis(ethylamine) as the N source and DL-malic acid as the
carbon precursor. Malic acid (1.5 g) was added in ∼3 mL of
2,2′-(ethylenedioxy)-bis(ethylamine), and then the mixture
was transferred into a 100 mL Teflon-lined stainless steel
autoclave. After keeping the mixture at 150 °C for 6 h and
cooling down to room temperature naturally, a red-brown
solution was obtained. The N-CDs were collected via
centrifugation at 8000 rpm for 15 min. The QY was measured
by the following formula by taking the reference standard dye
quinine sulfate (QY ∼ 54%).
I A ∩ 2
Q = Qr × × r × 2
Ir A ∩r
Here, Q is the quantum yield, I is the integrated emission
intensity, ∩ represents the solvent refractive index (1.33 for
water and 1.34 for 0.5 M H2SO4), A is the optical density, and
the subscript “r” indicates the reference dye. Quninine sulfate
was dissolved in 0.5 M H2SO4, whereas N-CDs were dissolved
in water.
4.3. Instrumentation. The morphological characteriza-
tions were performed with TEM and HRTEM using a Tecnai
20 G2 electronic microscope with an accelerating voltage of
300 kV. Samples for TEM/HRTEM were prepared by placing
a small drop of aqueous suspension of N-CDs onto a carbon-
coated copper grid and following solvent evaporation in a
vacuum oven. The UV−vis absorption spectra were obtained
at room temperature with a PerkinElmer Lambda 35
spectrometer. Photoluminescence spectral analysis was per-
formed in aqueous solutions at room temperature with a
PerkinElmer LS55 spectrophotometer. FT-IR spectra were
obtained using pressed KBr pellets with a Bruker Vertex 70
FT-IR spectrophotometer. XPS measurements were carried
out using ESCA+ omicron nanotechnology, Oxford Instru-
ments.
4.4. FL Detection of Cr(VI) and AA. The detection of
Cr(VI) and AA using N-CDs was performed in DI water at
room temperature. The FL intensity of N-CDs was analyzed at
320 nm excitation wavelength and denoted Io (FL intensity at
the initial point). Following that, 400 μL of Cr(VI) solution (5
mM) was added to the N-CD solution (2 mL) progressively in
26 steps. The final volume of mixtures was maintained to 2 mL
at room temperature, and FL spectra were obtained after 2 min
of incubation. The selectivity of Cr(VI) was assessed in
triplicate under the same experimental conditions on the basis
of variation in FL intensity using Cr(VI), Bi(II), Cu(II),
Ni(II), Co(II), Sr(III), Cr(III), Zn(II), and Hg(II). The
detection of AA was assessed by the FL recovery of the N-CD
solution, quenched with Cr(VI). Typically, 0−350 μL of AA
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solution (5 mM) was added to the N-CD−Cr(VI) mixture in
progressively 21 steps. In all of the experiments, final volumes
were maintained to 2 mL. The interference effect of other
biomolecules, such as Ad, Cy, Dt, Gu, Pr, SA, Th, and Gl, was
analyzed individually under the same experimental conditions.
All of the FL spectral measurements were performed at fixed
excitation and emission slits.
4.5. Antibacterial Test. E. coli and S. aureus cells were
grown separately in Lysogeny broth media. The antibacterial
assay for N-CDs against different bacterial strains was
performed in 96-well plates. E. coli and S. aureus were
incubated separately with varying concentrations of sterilized
N-CDs (10, 25, 50, and 75 μL mL−1 of media) at 37 °C for 12
h in a dark incubator shaker. The optical densities of the
samples with respect to the control were measured at 600 nm
excitation.
■ AUTHOR INFORMATION
Corresponding Author
*E-mail: sksonkar.chy@mnit.ac.in.
ORCID
Gunture: 0000-0001-5293-9173
Sumit Kumar Sonkar: 0000-0002-2560-835X
Notes
The authors declare no competing financial interest.
■
ACKNOWLEDGMENTS
S.R.A. thanks DST, New Delhi, for funding; A.B. thanks MNIT
Jaipur for a doctoral fellowship. D.S. thanks DST Inspire
doctoral fellowship. G. thanks CSIR for senior research
fellowship, and P.K. thanks CSIR (project no. 01(2854)/16/
EMR-II) and S.K.S. thanks DST (SB/EMEQ-383/2014) and
CSIR (01(2854)/16/EMR-II) for funding. S.K.S. thanks
Material Research Centre (MRC), MNIT Jaipur, for material
characterizations. S.K.S. thanks to Sankalp Verma and Dr.
Vivek Verma from Indian Institute of Technology, Kanpur for
the bacterial experiment.
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1591 DOI: 10.1021/acsomega.8b03191

ACS Omega 2019, 4, 1581−1591