Journal of Materials Sciences and Applications

2017; 3(6): 83-93

http://www.aascit.org/journal/jmsa
ISSN: 2381-0998 (Print); ISSN: 2381-1005 (Online)

Single Step Synthesis of Carbon

Quantum Dots from Coconut Shell:
Evaluation for Antioxidant Efficacy
and Hemotoxicity
Lakshmi Adinarayana Avinash Chunduri1, Aditya Kurdekar2,
Sandeep Patnaik1, Rajasekhar3, Saikiran Aditha2,
Chinnakota Prathibha2, Venkataramaniah Kamisetti2, *
1
Andhra Pradesh Medtech Zone, Vishakhapatnam, Andhra Pradesh, India
2
Laboratories for Nanoscience and Nanotechnology Research, Sri Sathya Sai Institute of Higher
Learning, Prasanthinilayam, Andhra Pradesh, India
3
Keywords Department of Physics, Rayalaseema University, Kurnool, Andhra Pradesh, India
Coconut Shell, Email address
Carbon Quantum Dots, [email protected] (V. Kamisetti)
Antioxidant Efficacy, *
Corresponding author
Hemotoxicity,
pH Sensor Citation
Lakshmi Adinarayana Avinash Chunduri, Aditya Kurdekar, Sandeep Patnaik, Rajasekhar, Saikiran
Aditha, Chinnakota Prathibha, Venkataramaniah Kamisetti. Single Step Synthesis of Carbon
Quantum Dots from Coconut Shell: Evaluation for Antioxidant Efficacy and Hemotoxicity.
Received: March 28, 2017 Journal of Materials Sciences and Applications. Vol. 3, No. 6, 2017, pp. 83-93.
Accepted: September 7, 2017
Published: November 25, 2017 Abstract
In this paper we report green synthesis of water soluble, monodisperse, and highly
fluorescent carbon quantum dots (CQDs) from coconut shell, a natural biomass as a
carbon precursor using a single step hydrothermal carbonization. These CQDs exhibited
blue fluorescence over excitation wavelengths of 300–380 nm and green luminescence
over excitation wavelengths of 400-490 nm. Structural and morphological
characterization were carried out using TEM, XRD, Raman and FTIR. As synthesized
CQDs were in the size range of 3 to 5 nm as observed in the TEM, while XRD results
confirmed their crystalline nature. Raman spectroscopic measurements exhibited the
characteristic “D” (1380 cm−1) and “G” (1596 cm−1) bands confirming the formation of
low dimensional carbon nanostructures. Optical properties of these CQDs were probed
using UV-visible spectrometry and room temperature Photoluminescence measurements.
The UV-visible spectrum showed a strong absorption at 283 nm. FTIR spectroscopy
confirmed the presence of hydroxyl and carboxyl functional moieties on the surface of
the CQDs, which play a crucial role in surface passivation leading to more stable
nanoparticle dispersions. The CQDs exhibited pH sensitivity over a wide pH range of 4–
11. This property can be exploited to develop a fluorescence based pH sensor. The high
antioxidant activity achieved using low concentration of CQDs (EC50=25 µg/mL) could
find potential applications in the treatment of various cancers and also food packaging.
Less than 5% hemolysis suggests that these CQDs are biocompatible and can be used for
various in vivo applications.

1. Introduction
Carbon Quantum dots (CQDs), a new member of carbon nanomaterial family was first
reported in 2004 by Scrivens [1]. These were considered to be clusters of carbon atoms
with diameters of typically 2 to 8 nm [2]. Although CQDs, Diamond nanocrystals, and
Graphene quantum dots are three similar quantum confined fluorescent carbon materials
84 Lakshmi Adinarayana Avinash Chunduri et al.: Single Step Synthesis of Carbon Quantum Dots from
Coconut Shell: Evaluation for Antioxidant Efficacy and Hemotoxicity

but the different spatial arrangements of carbon atoms result in scavenging the free radicals which are generated due to
in distinctive physical and chemical properties [3]. CQDs are interaction of biomolecules with molecular oxygen (O2)
mainly composed of sp2 carbon, oxygen, and nitrogen during metabolism. These free radicals or reactive oxygen
elements and other doped heteroatoms [4]. They do not species are main cause for diseases such as atherosclerosis,
measurably swell in aqueous solution but aggregation was Alzheimer’s and Parkinson’s, along with many cancers and
occasionally observed. The advantage of the CQDs is that other resulting effects of aging. Other members of carbon
they can be made strongly fluorescent without any doping. nanomaterial family have been already studied for their
We can also prepare functionalized CQDs in a single step [5]. antioxidant properties and they also exhibited good
CDs are generally regarded as a potential candidate in bio- scavenging activity [32], [33], [33]. Here we report one step
sensing, bioimaging, and other biologically related green synthesis of CQDs from agricultural waste i.e., coconut
applications due to water solubility, low cost, quantum yield, shell as the carbon source. We synthesized monodisperse,
low cytotoxicity and high chemical and photo stability [6], water soluble, highly fluorescent blue emitting CQDs by
[7]. The emission of CQDs can be tuned by varying the single step hydrothermal carbonization from coconut shell.
experimental conditions of synthesis to some level. For These CQDs were explored for their antioxidant activity by
CQDs the occurrence of both excitation and emission spectra DPPH assay and hemolysis assay for their cytotoxicity.
are very wide and their emissions vary from blue to red [8].
These wide emissions practically eliminate them for 2. Experiments
multiplexing assays. The quantum yield of CQDS ranges
from 5-45% this yield can further be improved by surface 2.1. Chemicals
passivation of metal ion doping [9]. Unlike semiconducting
quantum dots the emission of these dots is pH dependent. All chemicals were of analytical grade and were used
They also exhibit excitation wavelength dependent emission without any further purification. Coconut shell was
depending on the surface trap sites present on CQDs [10]. purchased from local market, washed and dried before use.
They also don’t have the blinking nature like semiconducting Millipore water was used throughout all the experiments.
quantum dots. During the past few years’ significant progress Quinine sulphate, tri sodium citrate and methanol were
has been made to prepare CQDs by various synthetic purchased from Merck. DPPH (2,2-diphenyl-1-
protocols such as arc discharge, laser ablation, thermal picrylhydrazyl- hydrate), ascorbic acid, Sodium borohydride,
cracking of organic compounds, electrochemical oxidation, phenol red, Iron (III) chloride, PBS (Phosphate Buffer Saline
microwave mediated synthesis, pyrolysis and oxidation of pH= 7.4), Triton- X-100 were obtained from Sigma-Aldrich.
candle soot [11]. Hydrothermal carbonization is a single step 2.2. Synthesis of Carbon Dots from Coconut
green synthesis procedure for the synthesis of carbon dots. Shell
Hydro thermal synthesis also has flexibility to choose over
a range of available precursors for the carbonization. Carbon dots were synthesized using locally available
Researchers have studied many natural resources such as soy agricultural waste i.e., coconut shell by hydrothermal
milk [12], meat [13], beverage [14], beer [15], egg [16], treatment in a single step. Coconut shell was grounded as
potato [17], punica granatum fruit [18], bombymori silk [19], powder and passed through 0.5 micron sieve to get uniform
sugar, bread, jaggery [20], lysozyme [21], and grass [22]. sized precursor particles. In this hydrothermal synthesis, 2
CQDs have been used extensively metal ion sensing [23], gm. of coconut shell was added to 50 mL of double distilled
photo catalysis [24], optical limiting [25], LED technology water. A 200 mL of Teflon lined autoclave was used for the
[26], bio imaging [27], bio sensing [28], bio medicine and reaction. Then the mixture was transferred into a 200 mL
drug delivery [29] applications. Apart from this they have Teflon lined autoclave and sealed very tightly to maintain the
also been studied for antibacterial, anti-inflammatory, inbuilt pressure. The autoclave was heated at 200 °C for 3
photodynamic therapy and antioxidant applications. With the hours. The reaction vessel was allowed to cool to room
growing interest of CQDs for clinical applications it is temperature and the solution was subjected to centrifugation
important to evaluate the cytotoxicity of these new class of to remove the carbon mass. The resultant fluorescent solution
carbonaceous materials developed. was passed through 0.2 µm filter to remove micron sized
Hemolysis assay is a simple but very important assay to particles. The final solution was lyophilized to obtain solid
evaluate the interaction of nanoparticles with blood CQDs. These CQDs were dispersed in water at a
components that would be administered intravenously. concentration of 1 mg/mL (Figure 1) and stored at 4°C for
Hemolysis may occur when the red blood cell membrane is further characterization and all applications. Reduced state
ruptured and releases hemoglobin into blood plasma which carbon dots (r-CDs) with strong blue luminescence were
may lead to anemia, hypertension, renal toxicity, etc. [30]. prepared by reducing CQDs with sodium borohydride in an
CQDs possess the electron donating and electron accepting aqueous solution. To prepare reduced CQDs (r-CQDs), an
properties there by acting as antioxidants and prooxidants excess solution of sodium borohydride was added and the
[31]. reaction was continued overnight at room temperature. The
In biological systems antioxidants, play an important role excess reductant was removed by dialysis against ultra-pure
 Journal of Materials Sciences and Applications 2017; 3(6): 83-93 85

water for 48 h. This reduction reaction does not increase in µg/mL) of CQD samples as test samples. Ascorbic acid (5
CQDs size. µg/mL) and methanol were chosen as positive control and
negative control respectively. These samples were incubated
in dark for 1 hour. After that the absorbance of the samples
was monitored at 517 nm for their antioxidant activity. The
DPPH scavenging percentage was calculated by the
following formula
%
100

where A represents absorbance in the respective controls.

2.5. Hemolysis Assay

The hemotoxicity of carbon dots was assessed by

performing the hemolysis assay. The assay was performed by
following the procedure reported by Miki et al. [42] with
minor modifications. 10 mL of human blood was collected
into a 3.8% citrate solution was centrifuged for 10 minutes.
The supernatant plasma was removed and erythrocytes
Figure 1. Synthesis of carbon quantum dots from coconut shell. (RBC) were washed multiple times with PBS (pH = 7.4).
These erythrocytes were diluted in 10 mL of PBS and the
2.3. Characterization hemolysis assay was performed by adding varying
The morphology and size of the CQDs was investigated by concentrations (25, 50, 100, 150 µg) of carbon dots to 1 mL
transmission electron microscope (TEM, JEOL, 2100F erythrocyte suspension placed in a 15-mL tube. Triton-X-100
electron microscope operated at 200 kV). The TEM sample and PBS were selected as positive and negative controls for
was prepared by drop casting diluted solution of CQDs on to this study. All the samples were diluted upto 10 mL with PBS
a carbon coated copper grid and dried under vacuum before and incubated at 37°C for 1 hour. Each test tube was gently
the analysis. Raman spectral measurements were carried out shaken to retain the contact of CQDs with blood components.
using Horiba Jobin Yvon, iHR 550 Raman spectrophotometer The samples were centrifuged to precipitate the RBC. The
equipped with 532 nm laser. FTIR spectrum was obtained on supernatant was collected and the absorbance was measured
a Shimadzu IR Affinity-1 FTIR spectrophotometer by KBr at 540 nm with a UV-visible spectrophotometer against PBS
pellet method. The spectrum was collected in transmission as blank. Hemolysis percentage was calculated using the
mode in the range of 400-4000 cm−1 with an accumulation of following formula.
32 scans and 2.0 cm resolution. Absorption spectra and
photometric studies were carried out on a Shimadzu UV- # $%& %
2450 UV-vis spectrophotometer.
where A represents absorbance in the respective controls.
2.4. DPPH Assay

Antioxidant efficacy of CQDs was measured ex vivo by 3. Results and Discussion

DPPH free radical assay method as reported by Berset et al.
[40] with slight modifications. DPPH is a dark-colored 3.1. Structural, Morphological and Optical
crystalline powder composed of stable free-radical Characterization
molecules. DPPH has major application in laboratory
In this study, we synthesized fluorescent carbon dots from
research for monitoring chemical reactions involving radicals
coconut shell in a single step by hydrothermal method. These
and it is the most common and popular antioxidant assay.
synthesized CQDs are soluble in water and also formed
DPPH acts as a free radical source and gets scavenged in the
stable suspensions.
presence of CQDs. DPPH is deep violet in color and it
becomes colorless or pale yellow over a period of time in the 3.1.1. TEM Analysis
presence of CQDs (antioxidants) when the DPPH radical gets The TEM image revealed the formation of CDs and their
neutralized. The change in the color is monitored for the monodispersed nature of CQDs synthesized from coconut
progress of the reaction and to measure quantitatively the shell. From figure (2a), the TEM image of carbon quantum
absorbance was measure at 520 nm. In this procedure 1 mL dots prepared from coconut shell reveals that they had
of (100 µM) methanolic DPPH solution was mixed with 1 uniform dispersion without apparent aggregation and all of
mL of various concentrations (10, 20, 50, 100, 200 and 300 them are spherical in shape with a narrow size distribution
86 Lakshmi Adinarayana Avinash Chunduri et al.: Single Step Synthesis of Carbon Quantum Dots from
Coconut Shell: Evaluation for Antioxidant Efficacy and Hemotoxicity

ranging from 3-5 nm. The formation of these small particles carbon quantum dots. Electron energy loss spectroscopy
with narrow size distribution is probably because of the (EELS) is a powerful tool to characterize the chemical
milder conditions applied for the synthesis. The high composition and structure of carbon-based materials. Figure
resolution TEM (HRTEM) image from figure 2(b) shows a 2 (d) shows the energy loss spectrum of the CQDs. The peak
crystalline nature with lattice space of 0.262 nm which at 285 eV is attributed to the transition from 1s to π* state
corresponds to the (002) diffraction facet of graphite [34]. (1s→π*), and the peak at 29 eV is the transition from 1s to
The SAED (figure 2(c)) pattern shows diffused rings σ* state (1s→σ*). These are the main EELS features of sp2
indicating the polycrystalline nature of the synthesized bonded (C=C) carbon at the K-edge region.

Figure 2. (a) TEM image of CQDs synthesized from coconut shell, (b) HRTEM image of CQDs showing lattice fringes (c) SAED pattern of CQDs (d) Electron
energy loss spectrum (EELS) of CQDS.

nature and is consistent with the TEM results.

3.1.2. Raman Analysis
Raman spectrum was employed to further characterize the
micro structure of CQDs synthesized from coconut shell.
Raman spectrum (Figure 3) displays two peaks at 1380 cm−1
and 1596 cm−1 which corresponds to disordered (D) band and
crystalline graphite (G) band respectively. The D band at
1380 cm−1 is related with the vibrations of dangling bonds in
the termination plane due to the presence of sp3 defects of
graphite. G band at 1596 cm−1 is related to in-plane vibration
of sp2 bonded carbon atoms which correspond to the E2g
mode in 2-D hexagonal lattice of graphite. These peaks are
characteristic of carbon dots and match well with the
literature values [35]. The ratio of intensities (ID/IG) of the
characteristic D band and G band can be used to correlate the
structural properties of carbon. The calculated ID/IG value is
1.1 for CQDs synthesized from coconut shell. The ID/IG less
than 2 indicates that CQDs have nanocrystalline graphitic Figure 3. Raman spectrum of CQDs derived from coconut shell.
 Journal of Materials Sciences and Applications 2017; 3(6): 83-93 87

3.1.3. Fourier Transform Infrared (FTIR) OH. The absorption peaks at 2923 and 815 cm−1 can be
Analysis assigned to the C-H stretching mode and C-H out-of-plane
bending mode. The peaks at 1600 and 1272 cm−1 are attributed
to the symmetric and asymmetric stretching vibration of
COO−, respectively. The Strong broad vibration around
1431cm−1 is associated with combined stretching due to
symmetric C-C and C-O vibrations. The presence of hydroxyl
and carboxyl groups might be due to the lignocellulosic
materials present in the coconut shell. The presence of the
hydroxyl and carboxyl (hydrophilic) groups on the surface of
CQDs imparts excellent water solubility and stability.
The same solution under UV lamp (365 nm) exhibited blue
emission shown in the inset of the figure 5. The optical
absorption peak of the carbon quantum dots was observed in
the UV region with maximum absorption at 283 nm. This
indicates that there is only one surface state present on the
CQDs for absorption. This is attributed to the n-π* transition
of C=O and π-π* transition of the conjugated C=C band
which are characteristics of the CQDs synthesized from
Figure 4. FTIR spectrum of CQDs derived from coconut shell.
biomaterials [36]. These synthesized CQDs are found to be
3.1.4. Optical Characterization stable for more than 6 months without any precipitation
As shown in the figure 4, the synthesized CQDs were which can be attributed to their small size and electrostatic
characterized for their functional groups by FTIR repulsions of the negatively charged functional groups
spectroscopy. The absorption peaks at 3300 cm−1 and 1050 present on the surface.
cm−1 corresponded to stretching vibrational absorptions of C-

Figure 5. UV-vis spectrum of CQDs synthesized from coconut shell and inset with photographs under white light (brown color) and UV lamp 365 nm (blue
color).

The classic signature of carbon dots is emission for all the excitations are located at 458 nm. It is interesting
wavelength and size dependent photoluminescent behavior. to note that there is no shift in the emission wavelength of the
From the fundamental as well as application viewpoint, PL is for the excitation upto 380 nm. From figure 7 a strong PL
one of the most fascinating behaviors of carbon dots. The PL emission peak located at 510 nm was observed with an
spectra of CQDs with variation in excitation wavelength excitation wavelength of 400 nm. The emission peak shifted
(300-380 nm and 400-490 nm) are shown in figure 6 and 7. to higher wavelengths with increase in excitation wavelength
When the excitation wavelength increases from 300 nm to but also decrease in the intensity was also observed.
380 nm the intensity also increased and the emission maxima The mechanism of the PL behavior is very complicated
88 Lakshmi Adinarayana Avinash Chunduri et al.: Single Step Synthesis of Carbon Quantum Dots from
Coconut Shell: Evaluation for Antioxidant Efficacy and Hemotoxicity

and there are no clear reports on the mechanism of the PL double distilled water. The absorbance of both the reference
behavior of carbon quantum dots till now. Two main reasons and samples was maintained close to 0.1 for avoiding any re-
for the PL behavior of CQDs are the presence of different absorption effects. The refractive index for both the reference
sizes and the distribution of the different surface energy traps and sample was n=1.33. The quantum yield was calculated
on the particles. The energy gap increases with the decrease using the following equation given [38] for both the
in size of the carbon dots and vice versa due to the quantum excitations (360 and 490 nm) of the CQDs.
confinement effect like semiconductor quantum dots. Thus, +
the particles with a smaller size get excited at a lower )* (
' '( +
wavelength, whereas those with a larger size get excited at )( * (
higher wavelengths. The intensity of the PL depends on the
number of particles excited at a particular wavelength. The
highest PL intensity of carbon dots was observed at an
excitation wavelength of 360 nm, due to the largest number
of particles being excited at that wavelength. Another reason
for the excitation dependent PL behavior of carbon dots is the
nature of their surface. The presence of various functional
groups on the surface of the carbon dots may result in a series
of emissive traps between π and π* of C-C. On illuminating
the carbon dots at a certain excitation wavelength a surface
energy trap dominates the emission. As the excitation
wavelength changes, other corresponding surface state
emissive traps become dominant. Hence the PL mechanism is
controlled by both the size effects and surface defects [37].

Figure 7. Photoluminescence emission spectra (Ex= 400 - 490 nm) of CQDs

synthesized from coconut shell.

where Q is the quantum yield, I is the integrated emission

intensity, n is the refractive index and A is the optical density and
subscripts R and S refer to the reference fluorophore of known
and unknown quantum yields. The fluorescence quantum yield
of these synthesized carbon quantum dots from coconut shell
calculated to be 35.74% for excitation at 360 nm using quinine
sulfate as reference (Table 1). When the CQDs were excited at
490 nm the quantum yield has reduced to 11.57% using
fluorescein as reference. This reduction in quantum yield can be
Figure 6. Photoluminescence emission spectra (Ex= 300 -380 nm) of CQDs
attributed to the less number of surface traps available at that
synthesized from coconut shell. excitation wavelength as explained in the previous section. The
observed quantum yields are higher than the literature reported
3.1.5. Calculation of Quantum Yield values for non-passivating carbon dots earlier. These high
The quantum yield of the synthesized CDs was calculated quantum yield CQDs can find their applications in bioimaging
by comparing the integrated emission intensities and and biosensors. In biological detection, these CQDs can be
absorbance values of quinine sulphate and fluorescein as applied at low concentrations as the PL intensity is sensitive to
reference. Quinine sulphate in 0.1M H2SO4 was chosen as a the concentration of carbon dots whereas at low concentration
standard with known quantum yield (QY) 0.54 at 360 nm. the variation of PL intensity is negligible.
Fluorescein in water was chosen as a standard with known
quantum yield (QY) 0.95 at 520 nm. CDs were dissolved in
Table 1. Quantum yield of the CQDs synthesized from coconut shell.

Description Fluorescein (490 nm) CQDs Quinine Sulphate (360 nm) CQDs
Integrated Emission Intensity 109697 12185 1981029 412042
Absorbance (A) 0.13 0.12 0.19 0.11
Refractive Index of the Solvent 1.33 1.33 1.33 1.33
Quantum Yield (Q) 0.95 0.11 (calculated) 0.54 0.193 (calculated)
Percentage (%) 100 11.57 100 35.7
 Journal of Materials Sciences and Applications 2017; 3(6): 83-93 89

3.1.6. CQDS for pH Sensor Applications

Figure 8 shows the variation of fluorescence intensity with
different pH values at an excitation wavelength of 380 nm. It
is evident that the emission intensities of our carbon quantum
dots synthesized from coconut shell are pH sensitive. The
emission maxima did not vary much as the pH is varied
(pH=4-11). A strong fluorescence was observed in the pH
range of 4-6. The fluorescence intensity has decreased
linearly with increase in pH from 4-12 when exited at 360
nm (figure 9). Change of pH from acidic to alkaline might be
causing the protonation and deprotonation of the carboxyl
groups which results in electrostatic charging of the CQDs
finally resulting in the Fermi level shift. The electronic
transitions of π–π* and σ-σ* are changed by refilling or
depleting their valance bands with the variation in pH. Even
though its mechanism is not fully understood this optically
pH sensitive property could be exploited for the fluorescence Figure 8. Fluorescence spectra of CQDs with variation in pH.
based pH sensors.

Figure 9. Variation of fluorescence intensity with pH when excited CQDs at 360 nm.

increased by 6 folds. From the figure 11 it is very clear when

3.1.7. PL Spectrum of r-CQDs the r-CQDs were excited with 420 nm also their fluorescence
In the present study sodium borohydride was specifically intensity increased by 4.5 times. A slight blue shift was also
chosen to reduce selectively the carbonyl and epoxy moieties observed in both the emission spectra. Since hydroxyl groups
to hydroxyl groups and significantly increase the amount of are electron-donating, so increasing their quantity by
hydroxyl groups on the CQDs without reducing other species reduction can enhance the luminescence of CQDs. Huzhi et
(C=C and COOH). Since there are large number of carboxyl al. [39] demonstrated the role of hydroxyl groups in the
groups present on the CQDs which was also confirmed by luminescence enhancement of r-CQDs. They treated the r-
FTIR, upon the reduction by sodium borohydride resulted in CDs with several strong oxidants like KMnO4, KIO4 and
higher number of hydroxyl groups on the surface and K2Cr2O7 that selectively oxidized hydroxyl groups and
increased the luminescence of the CQDs. The CQDs also resulted in quenching of luminescence of r-CQDs. They also
remained well dispersed even after reduction. As shown in treated with other oxidants like H2O2 and KBrO3 but they
figure 10 When the r-CQDs were excited with 380 nm could not quench luminescence of r-CQDs since they can’t
resulted in highly luminescent CQDs. Their intensity also oxidize hydroxyl groups.
90 Lakshmi Adinarayana Avinash Chunduri et al.: Single Step Synthesis of Carbon Quantum Dots from
Coconut Shell: Evaluation for Antioxidant Efficacy and Hemotoxicity

Figure 10. Comparison of fluorescence intensity of CQDs before and after reduction with NaBH4 (EX = 360 nm).

Figure 11 Comparison of fluorescence intensity of CQDs before after reduction with NaBH4 (EX = 420 nm).

3.2. Evaluation of Antioxidant Efficacy of CQDs

Even though there have been many reports available on the oxidative stress induced by many nanomaterials, carbon
nanomaterials were considered to be exceptional due to their non-toxic intrinsic carbonaceous nature. DPPH assay is a
standard method to measure the antioxidant activity of any compound used in industry. This assay provides an easy and rapid
way to evaluate antioxidant activity of any material.
 Journal of Materials Sciences and Applications 2017; 3(6): 83-93 91

Figure 12. DPPH-free radical scavenging assay of CQDs derived from coconut shell.

Antioxidants help prevent the damaging effects of

oxidation on cells throughout the body and nanomaterials
with antioxidant and antibacterial capabilities would find
extensive applications clinically in the preparation of
bandages for wound healing [41]. In the present It is
interesting to note that the antioxidant activity increased
exponentially with very small changes in the concentration.
EC50 is defined as the concentration at which 50% of the
desired effect is observed in the system. From the Figure 12
EC50 value for CQDs was calculated to be 25 µg/mL. This
high scavenging activity could be due to the large number of
electron donating groups (carboxyl and hydroxyl) present on
the surface.
3.3. Hemotoxicity Analysis of CQDs

Hemolysis assay was performed to evaluate the

applicability of the synthesized CQDs for in vivo system
such as optical imaging of blood and other blood related
sensing protocols. The in vitro hemolytic tests were often Figure 13. Percentage of cell hemolysis for various concentrations of CQDs
from coconut shell.
performed in pharma industry to screen drugs in their early
clinical development. It is a simple and reliable procedure to 3.4. Conclusion
estimate the biocompatibility of nanomaterials [43]. From
figure 13 the largest hemolysis percentage observed is 0.75% Carbon quantum dots were synthesized from coconut shell
with highest concentration 150 µg/mL of CQDs used in this in a single step by hydrothermal carbonization. Coconut shell
study. According to ASTM E2524-08 standard [44] (Standard is agricultural waste material which makes the production of
test method for analysis of hemolytic products of these CQDs to be cost effective. The presence of hydrophilic
nanoparticles), it has been reported that less than 5% groups such as carboxyl and hydroxyl groups present on the
hemolysis is permissible for biomaterials. From this study, surface confer them water solubility. These CQDs exhibited
we claim CQDs derived from the coconut shell as non-toxic blue fluorescence over excitation wavelengths of 300–380
quantum dots to RBCs even at higher concentrations. nm and green luminescence over excitation wavelengths of
400-490 nm. The CQDs exhibited pH sensitivity over a wide
92 Lakshmi Adinarayana Avinash Chunduri et al.: Single Step Synthesis of Carbon Quantum Dots from
Coconut Shell: Evaluation for Antioxidant Efficacy and Hemotoxicity

pH range of 4–11. The change in the fluorescence intensity detection application as nanoprobes. Nano-Micro Letters, 5
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fluorescence based pH sensor. It was elucidated the main Yan, and Minghong Wu. Observation of pH-, solvent-, spin-,
fluorescence is because of the hydroxyl groups which was and excitation-dependent blue photoluminescence from
confirmed by selectively reducing the carbon dots with carbon nanoparticles. Chemical Communications, 46 (21):
3681–3683, 2010.
sodium borohydride. The high antioxidant activity can be
achieved using low concentration of CQDs (EC50=25 [11] Jilong Wang and Jingjing Qiu. A review of carbon dots in
µg/mL) could find potential applications in the treatment of biological applications. Journal of Materials Science, 51 (10):
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soy milk and application as metal-free electro-catalysts for
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