Colloids and Surfaces A 586 (2020) 124266

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Colloids and Surfaces A

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

Ultra-radiant photoluminescence of glutathione rigidified reduced carbon T

quantum dots (r-CQDs) derived from ice-biryani for in vitro and in vivo
bioimaging applications
Anthony Ancy Milrada,1, Murugan Ramaduraia,1, Subramanian Raghunandhakumarb,
Selvarangan Gokul Krishnab, Pandurangan Prabhua,*, Dhanasekaran Anuradhab, Sohrab Abbasc
a
Department of Physical Chemistry, School of Chemical Sciences, University of Madras, Guindy Campus, Chennai, 600 025, India
b
Centre for Biotechnology, Anna University, Chennai, 600 113, India
c
Solid-State Physics Division, Bhabha Atomic Research Centre, Mumbai, 400 085, India

G R A P H I C A L A B S T R A C T

A R T I C LE I N FO A B S T R A C T

Keywords: Fabrication of beneficial materials from food waste is an extremely challenging task in the current research
Daily food waste scenario. In this study, we report the facile synthesis of fluorescent carbon quantum dots (CQDs) using ice-
Carbon quantum dots biryani (processed white rice waste) as a precursor and its application in bioimaging was evaluated using in vitro
Glutathione and in vivo models. CQDs was synthesized from carbonized ice-biryani by means of nitric acid treatment. To
Radiant photoluminescence
improve the quantum yield (QY) of as synthesized CQDs, we adopted two-step surface modification processes
High quantum yield
In vivo bioimaging
i.e., (i) preparation of reduced-state CQDs (r-CQDs) via borohydride reduction method followed by (ii) rigidi-
fying with a tripeptide i.e., glutathione (r-CQDs-GS).The effective interactions of oxygen rich r-CQDs and GS
stabilization has enhanced the QY from (bare CQDs) 5.46 % to (r-CQDs-GS) 41 %. The resultant r-CQDs-GS was
found to be an average particle size of ∼2−5 nm with pH tuned emission between pH 1–10. The biocompat-
ibility of r-CQDs-GS was tested using A549 cells and it was observed that almost 90 % cell viability and normal
morphology was retained even after treated with concentrations up to 100 μg/ml. The synergetic effect of r-
CQDs rigidified with glutathione have resulted in an efficient in vitro/in vivo bioimaging applications.

⁎
Corresponding author.
E-mail address: pprabhumu@gmail.com (P. Pandurangan).
1
These authors contributed equally to this work.

https://doi.org/10.1016/j.colsurfa.2019.124266
Received 8 September 2019; Received in revised form 21 November 2019; Accepted 22 November 2019
Available online 22 November 2019
0927-7757/ © 2019 Elsevier B.V. All rights reserved.
A.M. Anthony, et al.
1. Introduction
Food waste remains a global concern that transpires throughout the
food chain from farm to fork. The harsh effects of food and waste dis-
posal in landfills are undesirably affecting the natural resources [1],
human health [2] and also indirectly it is a chief source of greenhouse
gases [3], that frequently emits methane and results in almost 28 times
grander global warming prospective than CO2 emission [4]. According
to the United Nations (UN) Food and Agricultural Organization (FAO),
one third of the food made for human consumption get wasted in nu-
merous ways and ∼8 % of all greenhouse gases originates from those
waste [1,4,5]. Especially in India, as per UN Development Programme
(UNDP) 40 % of the food produced was wasted [6] and Agricultural
Ministry reports suggest 50,000 crore worth of food produced spews out
every year [7]. Therefore, the conversion of food waste into a beneficial
materials could be an innovative method to address this issue. In this
respect, research on developing bio-products such as ethanol, hy-
drogen, biodiesel, etc., from food waste have been well established
[8–12].
Recently, the luminescent Carbon quantum dots (CQDs) are suc-
cessfully fabricated from the food waste and numerous waste materials
obtained from diverse sources [4,13,14]. CQDs have become an emer-
ging class of quasi-spherical fluorescent materials with size less than
10 nm and composed of sp2 hybridized carbon with oxygen rich func-
tional groups such as hydroxyl, carboxyl, carbonyl and epoxy groups at
the surface [15,16]. Fluorescent CQDs possesses unique properties such
as low toxicity, good aqueous solubility, low cost, biocompatibility,
facile modification, tunable photoluminescence (PL) property, excellent
stability and great resistance to photobleaching, consequently it finds as
an alternate candidate to replace semiconductor quantum dots and
metal nanoclusters [17–21]. In particular the Cadmium (Cd) based
semiconductor QDs involving Cd possess high QY and tunable fluor-
escence property, still the nature of toxicity level and photobleaching
property of these QDs even at trace levels finds less significance for in
vivo bioimaging applications [22–27]. The metal nanoclusters (e.g., Ag
and Au nanoclusters) possess good PL property and also act as a safer
substitute for the purpose of bioimaging but they are highly expensive
and lack photostability [28–30]. Hence, the CQDs processed from less
expensive and biocompatible sources finds greater biological applica-
tions. Sarswat et.al and Park et.al previously reported the synthesis of
luminescent CQDs from food waste with the QY of 0.26 % and 2.85 %
respectively [4,13]. Recently, numerous attempts have been made to
enhance the QY and PL property of CQDs, via surface reduction [31]
and hetero atom doping [32–34]. Surface reduction enables the de-
duction of the non-radiative electron-hole recombination centers such
as epoxy and carboxylic acid groups that in turn increases the QY [18]
and doping with multi hetero atoms will induce and create additional
active sites on the surface of CQDs and thereby generated unique PL
properties [35]. Zheng et.al, first reported the reduction of CQDs using
NaBH4 and thereby enhanced the fluorescence QY from 2 % to 24 %
[31]. Hu et.al reported, the synthesis of fluorescent N and S-doped
CQDs using rice as the precursor and N-acetyl-L-cysteine as stabilizer
resulted in a QY of 2.36 % [36]. Most recently, Wang et.al, described
the synthesis of N, S-doped CQDs using ascorbic acid and ammonium
persulphate as starting material and achieved a maximum QY of 33 %
[37].
Glutathione (GSH) is a thiolated tripeptide (γ-glutamylcystei-
nylglycine), composed of three different amino acids such as cysteine,
glycine and glutamate. GSH is known as the mother of all antioxidants
that activates our body to supply on its own to protect us from cancer
and DNA damaging oxidative stress and so on [38,39]. Numerous re-
ports are available in the literature using carbohydrate as a precursor
material for the synthesis of fluorescent CQDs [40–50]. For example,
Wang et.al reported the synthesis of surface passivated CQDs (glucose
as carbon precursor) using GSH resulting in QY of 7.2 % [43]. Recently,
QY of 12.7 % was achieved by GSH passivated CQDs using prickly pear
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Colloids and Surfaces A 586 (2020) 124266
cactus fruit as carbon source [51]. In this background, we report the
fabrication of reduced CQDs rigidified with GSH (r-CQDs-GS) using ice
biryani (processed white rice waste) as a precursor material and we
have enhanced the QY by borohydride reduction followed by stabili-
zation with GSH. In comparison with reported methods [31,52,53], we
have used very low concentration of HNO3 (1 M) for the fabrication of
r-CQDs-GS at room temperature. As achieved r-CQDs-GS has exhibited
an ultra-radiant PL property for both in vitro and in vivo bioimaging
application was discussed hereafter.
2. Experimental section
2.1. Materials and characterization
Nitric acid (69 %, MERCK), L-Glutathione reduced (L-GSH, 99 %,
SRL chemicals extra pure), sodium borohydride and all other chemicals
utilized were of AR grade and used as received without further pur-
ifications. UV–vis absorption spectra were recorded using UV–vis
spectrophotometer, Perkin Elmer Lamba 650 and the photo-
luminescence spectra were recorded using Fluoromax Plus
Spectrofluorimeter. Fourier-transform infrared (FT-IR) spectra were
recorded using Bruker instrument in the range of 400 to 4000 cm−1.
Fluorescence life-time measurements were carried out using Time
Correlated Single Photon Counting (TCSPC), Fluorocube life-time
system. The samples were excited using 340 nm LED. Zeta potential
measurement was done using Horiba SZ-100.
2.2. Synthesis of r-CQDs-GS
The synthesis of r-CQDs-GS from ice-biryani involves three steps as
shown in the Scheme 1. Briefly in the Step 1 about 1 kg of the ice-
biryani (processed white rice waste) was collected from residential
source and washed thoroughly with hot water to remove the impurities
followed by dried under open air and crushed to a fine powder con-
sistency using mortar and pestle. The resulting fine powder were car-
bonized using muffle furnace for about 24 h at 250 ̊C. After purification
with acetone for three times the carbonized powder (2 g) was kept in a
100 ml flat bottom flask and 50 ml of 1 M HNO3 was added followed by
stirring for 36 h at 1200 rpm. After stirring a yellow color supernatant
along with black precipitate was obtained. The supernatant liquid was
collected by high speed centrifugation and filtered using 0.2 μm PTFE
filter and subjected to dialysis after adjusting the pH to neutral condi-
tion using 0.1 M NaOH solution. Step 2: In order to improve the QY as
well as photoluminescence property of as prepared CQDs, as suggested
by Zheng et.al [31] the above CQDs was reduced with 30 mg of NaBH4
added to the aqueous solution of CQDs (1 × 10−5 g/ml, 100 ml) and
stirred for an optimized 2 h resulted in r-CQDs. Step 3: After that about
30 mg of glutathione (GSH) was added to 10 ml of r-CQDs solution and
stirred for an optimized 2 h at ambient temperature. The obtained r-
CQDs-GS was subjected to further purifications using high speed cen-
trifugation followed by effective filtration with 0.2 μm PTFE filter and
dialyzed. The resulting r-CQDs-GS was used for further investigations.
2.3. QY calculation
The QY of the CQDs was calculated by relative method i.e., by
comparing the integrated photoluminescence (PL) intensities (excited
at 366 nm) and the absorbance values (at 366 nm) of the CQDs using
quinine sulphate (QS) as a reference. The reference was dissolved in
0.1 M H2SO4 and its QY was reported as 54 % (ƞ = 1.33). The solvent
used to dissolve CQDs was water (ƞ = 1.33). In order to reduce the re-
absorption effects, the absorbance was kept below 0.05 at 366 nm. The
integrated PL intensity is the area under the PL curve in the wavelength
range from 376 to 600 nm. The QY was calculated using the Eq. (1),
A.M. Anthony, et al.
Scheme 1. Pictorial representation of the steps involved in the synthesis of fluorescent r-CQDs-GS from daily food waste.
2
A ODr ⎞ ⎛ ηs ⎞
φs = ⎛ s ⎞ ⎛
⎜ ⎟⎜ ⎜
⎟ ⎟ φr °C
⎝ r ⎠ ⎝ ODs ⎠ ⎝ ηr ⎠
A (1)
Where, φ is the quantum yield, A is the integrated PL intensity, OD is
the optical density and η is the refractive index of the solvent used. The
subscripts ‘s’ and ‘r’ refers to sample and reference with known QY
respectively
2.4. The cytotoxicity test of CQDs
The cytotoxicity of CQDs (bare CQDs, r-CQDs and r-CQDs-GS) were
investigated using MTT assay in A549 (Human Lung Carcinoma) cell
lines. Briefly, A549 cells were seeded into a 96-well plate at a density of
2 × 104 cells per well and maintained in high glucose Dulbecco’s
modified Eagle’s medium (DMEM) containing 10 % fetal bovine serum
(FBS), 1 % antibiotics (penicillin/streptomycin), 1 % non-essential
amino-acid at 37 °C and 5 % CO2. After 24 h treatment with different
concentrations (0−100 μg/ml) of CQDs the cells were washed with 1x
PBS (pH = 7.4) and the plate was incubated with 50 μl of MTT [3-(4, 5-
Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide] solution
(final concentration 0.5 mg/ml) for 4 h. Further, the MTT medium was
replaced with 100 μl of DMSO to dissolve the formazan crystals. The
intensity of the dissolved formazan was measured by microplate reader
at wavelength of 492 nm and 650 nm (Bio Tek instruments, USA).
2.5. In vitro bio imaging
The A549 cells (4 × 104 cells per well) were seeded in 12 well plate
and incubated until they reach ∼70 % confluence with intact normal
morphology. Further, the cells were treated with CQDs (bare CQDs, r-
CQDs and r-CQDs-GS) (100 μg/ml) in DMEM medium at 37 °C for 1 h
with respective experimental designs. After incubation, the cells were
washed with 1x PBS solution (pH = 7.4) to remove the extracellular
CQDs from the plate. Subsequently, the plate was observed under
fluorescence microscopy at 488 nm using Invitrogen™ EVOS™ FLoid™
Cell Imaging Station (Fluorescent microscope)
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Colloids and Surfaces A 586 (2020) 124266
2.6. In vivo bio imaging
The in vivo tracking of r-CQDs-GS experiment were designed and
conducted according to the regulation of the institutional animal ethical
committee (IAEC No.CBT/AU/01/17). About 100 μl of r-CQDs-GS
(50 nmol) solution was administrated into swiss albino mice by tail vein
injection using a 0.5 ml insulin syringe. The in vivo bioimaging images
were recorded using Calipers IVIS Lumina LT series-III model with
Perkin Elmer live image 3.2 version software.
3. Results and discussion
3.1. Synthesis of r-CQDs-GS
The facile synthesis of r-CQDs-GS was carried out using ice biryani
(processed white rice waste) as carbon precursor. After carbonization
the fine powder of processed white rice waste was purified with acetone
to remove adsorbed impurities followed by nitric acid treatment. By
this treatment the large hydrophobic carbonaceous material was re-
duced into nanosized particles rich in oxygen-active functional groups
such as hydroxyl, carbonyl and carboxyl moieties. These surface groups
are responsible for the hydrophilic nature and negatively charged sur-
face of CQDs [31,54,55]. The steps involved during the treatment of
carbonaceous material with nitric acid are 1) narrowing the bulk
carbon aggregates into tiny nanosized particles 2) to solubilize these
tiny nano particles 3) thereby enhance the fluorescence properties of
CQDs and finally 4) generation of hydrophilic CeOeH and O]CeOeH
from hydrophobic CeH group [14,55]. Further, the carbonyl groups
present in the surface of CQDs was reduced using NaBH4. The sig-
nificant role of NaBH4 is to reduce the carbonyl and epoxy groups to
hydroxyl groups and leaving C]C and eCOOH groups ideal thereby
enhance the hydroxyl moiety on the surface of CQDs resulting in the
formation of r-CQDs as shown in step 1 of Scheme 2 [31]. Then the
surface modification with GSH over r-CQDs via the primary amine
group and thiol group present in GSH results in r-CQDs-GS (step 2 of
Scheme 2). The interaction of r-CQDs and GSH was optimized at 36 h
based on the maximum fluorescence response at this reaction time as
A.M. Anthony, et al. Colloids and Surfaces A 586 (2020) 124266

Scheme 2. Stepwise illustration of the surface modified CQDs responsible for the enhanced QY of r-CQDs-GS.

Fig. 1. (a) The HR-TEM image of r-CQDs-GS and inset shows (i) the lattice fringes, (ii) the corresponding selected area electron diffraction pattern (SAED) image of an
individual r-CQDs-GS and (iii) magnified HR-TEM image of r-CQDs-GS. b) FT-IR spectra of (i) r-CQDs-GS (ii) r-CQDs (iii) bare CQDs and (iv) control GSH respectively.

Fig. 2. (a) The absorption (black), excitation

(red), and emission (blue) spectra of r-CQDs-
GS. (Inset: aqueous solution of r-CQDs-GS
under daylight (i) and UV light (ii)) (b)
Excitation wavelength-dependent emission
spectra of r-CQDs-GS and (c) Comparative PL
spectra of bare CQDs, r-CQDs and r-CQDs-GS
(excited at 340 nm). (For interpretation of the
references to colour in this figure legend, the
reader is referred to the web version of this
article.)

shown in the Fig. S1 of electronic Supporting information (ESI).
3.2. Physicochemical properties
The structure and morphological features of r-CQDs-GS was ex-
plored by HR-TEM analysis as shown in Fig. 1a. The average particle
size of the r-CQDs-GS was calculated as ∼ 2−5 nm and the lattice
fringes (inset (i)) was measured as 0.31 nm which corresponds to the
(002) facet of graphite [56]. Besides, the selected area electron dif-
fraction (SAED) pattern image (inset (ii)) suggest the crystalline nature
of the as-synthesized r-CQDS-GS [34]. For better clarity, the magnified
HR-TEM image of r-CQDs-GS was also presented in the insert (iii). The
surface modification of GSH over r-CQDs was confirmed by the differ-
ence in the vibrational stretching patterns of r-CQDs-GS (i), r-CQDs (ii),
bare CQDs (iii) and control GSH (iv) was shown in the Fig. 1b. The
broad vibrational band between 3500 cm−1 to 3100 cm−1 corresponds

4
A.M. Anthony, et al. Colloids and Surfaces A 586 (2020) 124266

to the stretching vibrations of NeH and OeH [14]. The surface mod-
ification of r-CQDs with GSH was confirmed from the absence of
characteristic band for thiol (-SH) group in the range of 2600 to
2550 cm-1 [57]. The vibrational absorption band at 1637 cm−1 and
1680 cm−1, indicates the carbonyl groups on the surface of r-CQDs-GS
and GSH respectively. Upon reduction of bare CQDs the vibrational
absorption band of C]O at 1633 cm−1 was slightly shifted to
1642 cm−1, indicates the carbonyl groups on the surface of bare CQDs
were reduced to hydroxyl group [31]. The characteristic vibrational
band at 1400 cm−1 corresponds to the eCOOH group [58]. The above
results suggest that the distribution of carboxyl, amino and thiol func-
tional groups decorated on the surface of r-CQDs-GS. The zeta potential
measurement of the as-synthesized r-CQDs-GS was found to be
-27.6 mV that indicates the presence of negative charges on the surface
of r-CQDs-GS as shown in Fig. (S2).
3.3. Optical properties
As achieved N, S-modified r-CQDs-GS was found to be highly water
soluble. The UV–vis absorption, excitation and emission spectra of r-
CQDs-GS was shown in Fig. 2a in which an absorption shoulder was
observed at 340 nm, ascribed to the n-π* transition of surface groups
[15]. Analogous to previous reports [14,43], aqueous solution of r-
CQDs-GS exhibits a bright blue emission under 365 nm UV light (inset
(i)), and it seemed to be transparent and pale yellow under daylight as
shown in the inset (ii) of Fig. 2a. This blue emission of r-CQDs-GS arises
from the recombination of electron-hole pairs in the localized sp2 car-
bonic center buried within the sp3 matrix [59]. The excitation spectrum
of r-CQDs-GS shows an intense peak at 345 nm, obviously confirms the
surface excitations. As we recorded the PL spectra for the as-synthesized
r-CQDs-GS, at different excitations from 280 to 500 nm (Fig. 2b) con-
firms the excitation dependent emission behavior and red shifted as the
wavelength was increased. This red shift of PL spectra was obviously
shown in the corresponding normalized PL spectra (Fig. S3). The en-
hanced fluorescence intensity was observed when excited at 340 nm
i.e., at 432 nm with a large stokes shift of ∼92 nm. The excitation (λex)
dependent emission wavelength and intensity was due to the excitation
of different sized CQDs within the r-CQDs-GS ensemble [14]. Interest-
ingly, the fluorescence QY of r-CQDs-GS, r-CQDs and bare CQDs were
calculated to be 41 %, 27.6 % and 5.46 % respectively, and QY of r-
CQDs-GS was superior in comparison with reported literature as shown
in Table 1. Fig. 2c shows the comparative PL spectra’s of bare CQDS, r-
CQDs and r-CQDs-GS. Here, as compared to bare CQDs and r-CQDs the
enhanced emission intensity was observed for r-CQDs-GS indicating its
higher quantum yield. The reason behind the high QY could be due to
the synergetic effect of peripheral ligand i.e., GSH over the surface of r-
CQDs and heteroatom distribution on the surface of r-CQDs-GS.
3.4. Effect of pH and stability of r-CQDs-GS
The effect of pH, on the stability of as synthesized r-CQDs-GS was
examined at different pH ranging from 1 to 10. Interestingly, r-CQDs-GS
exhibited pH tuned emission responses as confirmed from the PL
spectra as shown in Fig. 3a. The results of calibration plot of PL in-
tensity versus pH (as shown in Fig. 3b) suggests that the emission in-
tensity reaches the maximum at pH = 4 but it decreases drastically
from the pH = 4 to pH = 8 and again increases from pH = 8 to
pH = 10. Meanwhile, a blue-shift in the emission peak from 438 to
432 nm was observed as pH increases from 1 to 4 and red shift from 432
to 446 nm (from pH = 4–10). This pH tuned emission behavior was due
to the N/S co-doped CQDs. The results also indicated that the newly
fabricated r-CQDs-GS was highly pH sensitive and also have a potential
to serve as proton sensors in the detecting cell metabolic process with
proton release [15]. The average life-time of r-CQDs-GS was found
using time-correlated single photon counting (TCSPC) technique as
shown in the Fig. 4a. The average luminescence life-time of r-CQDs-GS
was found to be 9.33 ns, which is superior as compared to r-CQDs
(7.61 ns) and bare CQDs (7.29 ns) that again confirms the enhanced PL
property of N/S co-doped r-CQDs-GS. The narrow difference in life-time
values of bare CQDs and r-CQDs needs further investigations to confirm
extraordinary role of GS in enhancing the QY upto 41 %. The storage
stability of r-CQDs-GS was examined by keeping the sample at room
temperature for 60 days (Fig. 4b) and effectively minor fluctuation in
the PL intensity and no precipitation of sample was observed indicating
their abundant stability and long shelf life.
3.5. Biocompatibility and bioimaging analysis
The biocompatibility of various CQDs is one of the most important
test to be exercised before it is introduced into biological application
[60]. To evaluate the biocompatibility properties of different CQDs
(bare CQDs, r-CQDs, r-CQDs-GS), we examined the cytotoxicity of CQDs
with different concentrations (0−100 μg/ml) on A549 cells using MTT
assay at 24 h drug treatment. The Fig. 5 shows, the 90 % cells retain the
normal morphology even at higher concentration (100 μg/ml) of CQDs,
when compared with untreated cells. It clearly demonstrates that the
non-toxic nature and excellent biocompatibility of different CQDs holds
promise for further experiments. As compared with reported literature
the CQDs shows low cytotoxicity even at higher concentration together
with biocompatibility and unique PL characteristic features to act as an
excellent bioimaging agent. Further, we explored the PL property of
CQDs using the live cell imaging of in vitro and in vivo analysis, which
was performed to reveal the biological applications of r-CQDs-GS. For in
vitro analysis (Fig. 6) the A549 cells were incubated for 1 h with bare
CQDs, r-CQDs and r-CQDs-GS separately. Interestingly, the r-CQDs-GS
has a higher PL efficiency as compared with bare CQDs and r- CQDs and
the images were captured at 488 nm (green fluorescence area) under
fluorescence microscope. Concomitantly, the r-CQDs-GS was taken for

Table 1
Comparison of synthetic conditions and quantum yields of CQDs obtained from various carbohydrate precursor.
S No Carbohydrate as precursor Dopant Preparation method Temperature (°C ) Time consumed QY (%) Ref

1 Glucose aq.NH3 Ultrasonication – 24h 6.65 [40]

2 Cyclodextrin – hydrothermal 70 4h 13.5 [41]
3 Fructose/Maltose – NaOH/ NaHCO3 RT 1h 2.2 [42]
4 Glucose Glutathione hydrothermal 180 22h 7.2 [43]
5 Glucose PEG-diamine Ultrasonication 40 4h 6.8 [44]
6 Glucosamine HCl Na4P2O7 Teflon-autoclave 180 10h 16.8 [45]
7 Cyclodextrin OEI hydrothermal 90 2h 30 [46]
8 Glucosamine HCl TTDDA microwave – 3 mins 18 [47]
9 Glucose EDA/conc.H3PO4 hydrothermal 130 6 mins 9.6 [48]
10 Cellulose Urea hydrothermal 200 72h 21.7 [49]
11 Glucose Ammonia and H3PO4 Autoclave reflux 160 5h 30 [50]
12 Ice-biryani (processed white rice waste) Glutathione Treated with 1 M HNO3 RT 36 h 41 This work

5
A.M. Anthony, et al.
Fig. 3. (a) The photoluminescence spectra (excited at 340 nm) of r-CQDs-GS at different pH and b) the corresponding calibration plot of PL intensity versus pH.
Fig. 4. (a) The fluorescence life time decay curve of r-CQDs-GS (green), r-CQDs (red) and bare CQDs (black) (b) PL spectra of r-CQDs-GS recorded over a period of 60
days. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
Fig. 5. Cell viability of CQDs (bare CQDs, r-CQDs, r-CQDs-GSH) and dose
fixation study was assessed by MTT analysis. A549 cells were treated with
various concentration of CQDs (0−100 μg/ml) for 24 h. The graph showing the
mean ± S.D. value of three independent experiments and significance
(p < 0.05) compared with the control group was indicated (*).
in vivo bioimaging (Fig. 7). The 100 μl (50 nmol) of r-CQDs-GS ad-
ministrated animal exhibits higher fluorescence signals as compared
with vehicle control alone injected animal. This clearly indicates that
the r-CQDs-GS has a unique characteristic feature like low cytotoxicity
and active biocompatibility with PL property even at different tem-
perature with different conditions are mainly associated with uniform
particle size, which help the r-CQDs-GS equal bio distributions in ex-
tracellular matrix and blood stream, upon in vitro and in vivo adminis-
tration [61]. Overall, the r-CQDs-GS have low cytotoxicity and active
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Colloids and Surfaces A 586 (2020) 124266
biocompatibility with unique fluorescence property could be a better
bio-labelling agent to targeted/encapsulated drug delivery and other
biological applications.
4. Conclusions
In summary, we have successfully fabricated r-CQDs-GS by a simple
method from ice-biryani (processed white rice waste) as a carbon pre-
cursor. As achieved r-CQDs-GS exhibits a maximum QY of 41 % by the
synergic role of reduced CQDs rigidified with GSH which was estab-
lished even under a very low concentration of HNO3 as compared with
reported literatures also the GS rigidified r-CQDs exhibits pH tuned
emission. The highly luminescent r-CQDs-GS was successfully demon-
strated for in vitro and in vivo bioimaging of A549 cells. Further, the
present results substantiates and promises the conversion of daily food
waste into cost-effective beneficial materials for the technological and
biomedical applications.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influ-
ence the work reported in this paper.
Acknowledgments
PP and AAM acknowledge the financial support rendered by UGC-
DAE-CSR (M-231), Bhabha Atomic Research Centre (Mumbai). PP and
MR acknowledge the Grants received from the Department of Science
and Technology (DST) - Science and Engineering Research Board
A.M. Anthony, et al. Colloids and Surfaces A 586 (2020) 124266

Fig. 6. Images of human lung cancer A549 cells

labeled with C-Dots (bare CQDs, r-CQDs and r-
CQDs-GS) obtained by a Invitrogen™ EVOS™
FLoid™ Cell Imaging Station (Fluorescent mi-
croscope). (a) Bright-field image; (b)
Fluorescent image. (40× objective), using a
405 nm laser and collecting the emitted fluor-
escence in the green area. (For interpretation of
the references to colour in this figure legend,
the reader is referred to the web version of this
article.)

Fig. 7. The in vivo image shows the Green r-

CQDs-GS accumulation inside the whole body.
The whole body fluorescent images taken 1 h
after tail vein injection shows the higher ac-
cumulation of Green r-CQDs-GS (488 nm), and
the red to flame yellow colour expressed the
higher intensity of fluorescence. (For inter-
pretation of the references to colour in this
figure legend, the reader is referred to the web
version of this article.)

(SERB) through Early Career Research Award (ECR) [SERB/ECR/2016/
000837; Dt:6/2/2017]. SR and DA acknowledge the funds received
from UGC, New Delhi (Kothari-PDF). We sincerely acknowledge the G.
N. Ramachandran (GNR) Instrumentation facility at University of
Madras, Guindy Campus. We also acknowledge the Sophisticated Test
and Instrument Centre, Cochin University of Science and technology
Cochin, for HR-TEM analysis.
Appendix A. Supplementary data
Supplementary material related to this article can be found, in the
online version, at doi:https://doi.org/10.1016/j.colsurfa.2019.124266.
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