Superlattices and Microstructures 83 (2015) 776784

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Superlattices and Microstructures

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

Few layered graphene Sheet decorated by ZnO

Nanoparticles for anti-bacterial application
Satish Bykkam a, Sowmya Narsingam a, Mohsen Ahmadipour b, T. Dayakar a,
K. Venkateswara Rao a,, Ch. Shilpa Chakra a, Shanker Kalakotla c
a
Centre for Nano Science and Technology, Institute of Science and Technology, Jawaharlal Nehru Technological University,
Hyderabad 500085, Telangana State, India
b
School of Materials and Mineral Resources Engineering, University Sains Malaysia, 14300 Nibong Tebal, Penang, Malaysia
c
Centre for Pharmaceutical Sciences, Institute of Science and Technology, Jawaharlal Nehru Technological University,
Hyderabad 500085, Telangana State, India

a r t i c l e

i n f o

Article history:
Received 18 March 2015
Accepted 19 March 2015
Available online 23 April 2015
Keywords:
Few layered graphene
ZnO nano particles
Antibacterial activity
Escherichia coli
Salmonella typhi

a b s t r a c t
A widely soluble few layered graphene (FLG) sheets decorated by
ZnO nano particles were prepared through hydrothermal method
using ethylene glycol as a solvent and a reducing agent. The
obtained FLG/ZnO composite material was characterized by X-ray
diffraction (XRD), Transmission electron microscopy (TEM) and
Raman spectroscopy. The anti-bacterial properties of ZnO nano
particles decorated few layered graphenes were tested against
Escherichia coli and Salmonella typhi by using well diffusion
method. The results conrmed that FLG/ZnO has signicant
antibacterial activity more against S. typhi than E. coli. The obtained
results from the current research work conclusively states that the
ZnO nano particles which were decorated by few layered graphene
possess a signicant anti-bacterial activity.
2015 Elsevier Ltd. All rights reserved.

1. Introduction
Microorganisms are everywhere in the biosphere, and their presence invariably affects the environment where they are growing in. The effects of microorganisms on their environment can be benecial
or harmful or in apparent with regard to human measure or observation. They are in natural
Corresponding author. Mobile: +91 9440858664.
E-mail address: Kalagadda2003@gmail.com (K. Venkateswara Rao).
http://dx.doi.org/10.1016/j.spmi.2015.03.063
0749-6036/ 2015 Elsevier Ltd. All rights reserved.

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777

symmetry with human body and environment. Therefore control of its harmful effects appears to be
most important criteria. To resolve this current problem physicians have recommended many techniques and drugs such as kanamycin, spectinomycin, and penicillin. However, frequent usage of these
agents causes the microbes to become resistant against those drugs. Nano materials have been to the
research focus of interest in the last decade. Recently strong antimicrobial activity has been revealed
by several nanomaterials. The metal and metal oxide nanoparticles such as silver (Ag), silver
oxide (Ag2O), titanium dioxide (TiO2), gold (Au), calcium oxide (CaO), silica (Si), copper oxide (CuO),
and magnesium oxide (MgO) have been reported to possess antimicrobial activity [17]. Eventually
the aqueous fullerene nanoparticles (nC60) [8] and carbon nanotubes (CNT) [9] have showed
antimicrobial activity.
Graphene is a two dimensional structure of carbon nanomaterials, it exhibit excellent mobility of
charge carriers and high surface area due to the presence of properties such as thermal, mechanical
and electrical [1013]. Outstanding and unique properties of graphene have many applications in
elds such as nanoelectronics [14], engineering nanocomposite materials [1521], energy storage
[2226], eld effect transistor (FET) [2729], sensors [30], catalysis [31] and biomedical applications
(biosensor, biodevices, drug and gene delivery, cancer therapy etc.) [32,33]. Additionally, the surface
properties of graphene could be modied via physical and/or chemical method, which extends its use
in composite materials [34].
Zinc oxide (ZnO) has an innate advantage of broad antibacterial activities against bacteria [35,36]
fungus [37,38] and virus [39]. Food and Drug Administration of the United States (21CFR182.8991)
had recognized zinc oxide as a safe material [40]. Thus, it has been widely used as an antibacterial
agent. In recent years, ZnO Nanoparticles (NPs) showed much better antibacterial ability than that
of the counterpart microsized ZnO particles. Release of zinc ions from ZnO was suggested as one of
the primary antibacterial mechanisms [41], moreover the penetration of a bacterial membrane upon
contact with ZnO particles also contributed to the antibacterial ability of ZnO NPs [42,43]. However,
the easy aggregation of ZnO NPs hinders their antibacterial activity [44]. Current research work
demonstrates the conversion of GO to FLG and determination of anti bacterial activity of few layered
graphene Sheet decorated by ZnO Nanoparticles.
2. Experimental
2.1. Materials
Graphite akes (300 meshes), Hydrochloric acid (HCl), Sulfuric acid (H2SO4), Hydrogen peroxide
(H2O2), Potassiumpermanganate (KMnO4), Zincacetate Zn(O2CCH3)2. Microbial Cultures: Escherichia
coli (E. coli) and Salmonella typhi (S. typhi), Nutrient Broth, Nutrient Agar, Antibiotic: Ampicillin,
Ethanol (changshu yangyuan).
2.2. Preparation of graphene oxide
Hummers method has been used to synthesize graphene oxide. In a typical synthesis procedure,
the graphite akes were added to a solution containing strong oxidizing agents (sulfuric acid and
sodium nitrate) and stirred for one hour. Potassium permanganate (KMnO4) was added slowly to
the mixture during addition the reaction kept at 0 C by using ice bath. Simultaneously the mixture
slowly shifted to another bottle which consists of additional water, further H2O2 was added to the
mixture. The color of mixture was changed to bright yellow indicating a high oxidation level of
graphite. The solution was ltered and washed several times with distilled water and HCl to remove
the remaining impurities. The washing process was carried out with a simple decantation of the
supernatant by centrifugation technique at 5000 rpm for 30 min which results in the formation of
graphene oxide (GO).

4KMnO4 2H2 O ! 4KOH 4MnO2 3O2

Cflake graphite nO2 ! GO

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2.3. Preparation of FLG/ZnO Composite

The puried graphene oxide (10 mg) was dissolved in 80 ml of ethylene glycol (EG) by Ultra
sonic treatment for 2 h under ambient condition to give a brown dispersion. Further 80 mg of zinc
acetate dissolved in 80 ml of EG and stirred to obtained brown dispersion. Subsequently, 20 mg of
NaOH dissolved in 20 ml of deionized water then the mixture was stirred for 60 min and
transferred to 250 ml glass bottle and tightly sealed with Teon cap then kept the glass bottle
in hot air oven for 10 h at 100 C. After completion of process, sample was cooled down to room
temperature. The obtained few layered graphene/ZnO in material was centrifuged and washed
several times with water and Ethanol thoroughly. The synthesized few layered graphene/ZnO
material was dried in oven at 700 C for 24 h. Schematic diagram and chemical formula of
preparation few layered graphene (FLG) sheets decorated by ZnO nano particles have been shown
in Fig. 1.

2.4. Well diffusion method

The sterilized nutrient agar was poured (20 ml/plate) onto the Petri plates & left for a while till the
agar gets solidied. Two pathogenic strains E. coli (gram negative) and S. typhi (gram positive) were
taken and made into cultures. Fresh overnight cultures of inoculum (100 ll) of each cultures
were spread on to nutrient agar plates surface evenly in four different petri plates respectively, the
wells were casted by porer (3 mm diameter). The samples along with standard antibiotic
(Ampicillin) were loaded with equal volume (30 ll) on the plates. Control plate consists of distilled
water and antibiotic drugs. The square plates were incubated at room temperature for 1924 h.
Activity was clearly visible from 19 to 24 h on the plates. Zone of inhibition was measured & the
sample of the FLG/ZnO showing maximum antimicrobial activity was noted.

Fig. 1. Formation mechanism of few layered graphene (FLG) sheets decorated by ZnO nano particles.

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779

3. Characterization
The crystal structure and phase of the sample were determined by X-ray diffraction (XRD), (Model:
Bruker D8 Advance, Germany). The surface morphology of the prepared FLG/ZnO examined by High
Resolution Transmission Electron Microscopy (Model: JEOLJEM-200CX), Raman spectroscopy were
recorded (Model of alpha 300 of WiTec, Germany).
4. Results and discussion
The FLG/ZnO composite was synthesized without any reducing agent by in situ reduction of zinc
acetate and graphene oxide at low temperature hydrothermal method. For buckling the huge amount
of ZnO Nanoparticles on the few layered graphene surface and as well as intercalated between the
sheets zinc acetate was favoured thus improving the physiochemical property of FLG/ZnO composites.
4.1. Crystal analysis
XRD pattern of graphite akes, graphene oxide and FLG/ZnO are shown in Fig. 2. Graphite akes a
strong and sharp peak at 2h = 29.4 is exhibited. Fig. 2(a), indicates a highly ordered structure corresponding to a basal spacing d002 = 0.334 nm. On the other hand the pattern of graphene oxide exhibits
a 0 0 1 reection at 9.8. Fig. 2(b) corresponds to a basal spacing of d001 = 0.961 nm. The interlayer
spacing of GO calculated was 0.961 nm. FLG/ZnO composites has ve main peaks at 2h = 32.6, 35,
36.8, 47.8, 56.5 which correspond to the (1 0 0), (0 0 2), (1 0 1), (1 0 2), (1 1 0) crystalline planes of
ZnO, respectively Fig. 2(c). The XRD patterns of FLG/ZnO showed one additional diffraction peak at
2h = 26.3 besides the peaks assigned to ZnO, which is the characteristic peak of few layered graphene.
4.2. Morphology
The High Resolution Electron Microscopy (HRTEM) image of the FLG/ZnO nanocomposite conrms
nanosized ZnO particles are uniformly decorated on graphene Sheet. A ne size distribution of around
3040 nm of quasi-spherical ZnO particles on graphene Sheet was exhibited. Almost all the ZnO
Nanoparticles were attached on graphene. The rm attachment of the ZnO particles was due to electrostatic attraction between the positively charged Zn2+ and the negatively charged graphene oxide
sheets. Fig. 3 provides the image of an individual ZnO particle on graphene. A good crystallization
of ZnO is observed in the HRTEM image which is in agreement with the XRD results. From the above

Fig. 2. XRD patterns of (a) Graphite akes, (b) graphene oxide and (c) FLG/ZnO composite.

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Fig. 3. TEM images of (ac) few layered graphene (FLG) sheets decorated by ZnO nano particles, (d) Selected Area Electron
Diffraction (SAED) pattern.

analysis it can be determined that a FLG/ZnO nanocomposite has formed by in situ on depot
hydrothermal process. Thus ZnO Nanoparticles were successfully decorated on the few layered
graphene Sheets with hexagonal wurtzite structure.
4.3. Raman spectrum
For investigative the ordered and disordered crystal structures of carbonaceous materials and to
identify the single-layer, bi-layer and multi-layer characteristics of graphene and/or graphene oxide
layers the Raman spectroscopy technique was employed. In Raman spectra carbon materials have G
line (1580 cm1) and D line (1350 cm1) which were usually allocated to the E2g [45,46] phonons
of SP2 carbon atoms and breathing mode of k-point phonons respectively showing local defects and
disorders of graphene and graphite platelets [47].
In Fig. 4(a) the two major peaks of Graphite akes at 1344 cm1 and 1575 cm1 indicate D-band
and G-band respectively. While two major Peaks at 1346 cm1 (the D-band) and 1580 cm1 (G-band)
was observed for graphene oxide as shown in Fig. 4(b). The graphite structure of GOs is indicated by G
band signature and the D-band signatures point to defects on the edges or surfaces of the GO. G band
is broadened and shifted to 1580 cm1 whereas the D band at 1346 cm1 indicating the creation of sp3
domains due to the extensive oxidation. FLG/ZnO Raman spectra in Fig. 4(c) showed two bands at
1345 cm1 and 1582 cm1 corresponding to the disordered (D) band and graphitic (G) band of carbon
materials. Wurtzite-type ZnO and the presence of carbon in FLG/ZnO are conrmed by the XRD and
Raman spectra respectively.

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781

Fig. 4. Raman spectrum of (a) Graphite akes, (b) graphene oxide and (c) FLG/ZnO composite.

5. Antibacterial activity
The antibacterial activity of the sample was determined by the formation of Zone of Inhibition.
Zone of Inhibition is the area on an agar plate where growth of a control organism is prevented by
an antibiotic usually placed on the agar surface. If the test organism is susceptible to the antibiotic,
it will not grow where the antibiotic is present. The size of the zone of inhibition is a measure of
the compounds effectiveness, the larger the clear area around the antibiotic, the more Effective the
compound.
The activity of the sample was observed by the formation of Zone of inhibition after 24 h. Presence
of zone of inhibition conrmed inhibitory activity of FLG/ZnO. The Zone of Inhibitions of different bacteria is given in Fig. 5. The control plates showed the growth of bacteria in the absence of antibacterial
agents. The clear zone surrounding the sample in the remaining plates shows the activity of the sample. Fig. 5 shows the petri dishes with samples of 0.01 g of FLG/ZnO and 0.05 g of FLG/ZnO. The zone
surrounding the sample is clear that shows complete zone of inhibition. The space surrounding the
complete zone of inhibition is partial zone of inhibition where the activity decreases than complete
zone of inhibition. The Zone of inhibition is more for the high concentration of FLG/ZnO.
In current research work from Fig. 5, the observations states FLG/ZnO nano powder has stronger
antibacterial activity for Gram positive than gram negative of E. coli. The concentration and zone of
inhibition of the S. typhi and E. coli are shown in histogram in Fig. 6. The S. typhi is a Gram positive
bacteria with thick cell wall consisting of a large number of mucopeptides, murein and lipoteichoic
acids. Moreover, the antioxidant enzyme of S. typhi yields a stronger oxidant resistance. Despite these
two reasons, a greater inhibition response is observed from S. typhi membrane has a smaller negative
charge than gram negative E. coli it would allow the penetration of a greater number of negatively
charged free radicals such as superoxide anions and hydroxyl radicals, into the cell membrane, causing
severe damage and death of the cell.
The results interpreted according to the standard protocols, may be related to several mechanisms
including the generation of reactive oxygen species (ROS) on the surface of the particles which allows
release of Zn2+ ions from the FLG/ZnO samples and the penetration of these nanoparticles which contributes to the mechanical destruction of cell membrane. The generation of ROS like OH, O2 and
H2O2 results, when the FLG/ZnO nanopowders are irradiated with light having high photon energy
or energy equal to the band gap, they cause transfer of electrons from the valence band to the conduction band of the product material. It leads to generation of holes in the valence band which can react
with hydroxyl groups and absorbed water to create hydroxyl radicals (OH). The electrons in the
conduction band can be trapped by the presence of O2 to produce superoxide radical anions (O2)

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Fig. 5. (a and b) Zone of Inhibition formation against Salmonella typhi FLG/ZnO (c and d) Zone of Inhibition formation against
Escherichia coli FLG/ZnO.

Fig. 6. Histogram shows the concentration and zone of inhibition of the S. typhi and E. coli.

which in turn can react with hydrogen ions to form HO2 radicals. The H2O2 can be generated by the
combination electron hole pair. The hydroxyl radicals and superoxide anions are negatively charged
and hence they cannot penetrate into the cell membrane, but they can cause fatal damage to the outer
surface of the bacteria like proteins, DNA and lipids, whereas, the H2O2 can penetrate directly into the
cell wall and it kills the bacteria. Table 1 shows the zone of inhibition diameter from this the increased
doping concentration is directly proportional to antibacterial zones.

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Table 1
Shows the antibacterial activity (Zone of inhibition) of S. typhi and E. coli.
S. No.

Organism

Zone of Inhibition (mm) at different concentrations (ll)

0.01 g

1
2

S. typhi
E. coli

0.05 g

25 ll

50 ll

75 ll

25 ll

50 ll

75 ll

7
6

9
8

12
9

8
7

10
9

13
11

6. Conclusion
Successfully Few layered graphene Sheet was decorated by ZnO Nanoparticles using hydrothermal
method. FLG/ZnO composite has shown maximum antimicrobial activity against E. coli and S. typhi.
From the results it has been concluded that the zone of inhibition increases as the concentration of
FLG/ZnO increased in both the bacteria. According to the results obtained the ZnO Nanoparticles
decorated by few layered graphene possess a good anti-bacterial activity.
Acknowledgements
Author sincerely thanks to University Grant Commission (UGC-Government of India) for nancial
support through Rajiv Gandhi National Fellowship (RGNF) Scheme. And the Centre for Nano Science
and Technology, Institute of Science and Technology JNTU Hyderabad for providing necessary facilities
to carryout present research work. And also thank to School of Materials and Minerals Resources
Engineering, University Sains Malaysia (USM), Malaysia.
References
[1] A. Azam, A.S. Ahmed, M. Oves, M.S. Khan, S.S. Habib, A. Memic, Antimicrobial activity of metal oxide nanoparticles against
Gram-positive and Gramnegative bacteria: a comparative study, Int. J. Nanomed. 7 (2012) 60036009.
[2] A. Besinis, T. Peralta, R.D. Handy, The antibacterial effects of silver, titanium dioxide and silica dioxide nanoparticles
compared to the dental disinfectant chlorhexidine on Streptococcus mutans using a suite of bioassays, Nanotoxicology 8
(2014) 116.
[3] Z. Emami-Karvani, P. Chehrazi, Antibacterial activity of ZnO nanoparticle on grampositive and gramnegative bacteria,
Afr. J. Microbiol. Res. 5 (2011) 13681373.
[4] M.S. Usman, M.E. El Zowalaty, K. Shameli, N. Zainuddin, M. Salama, N.A. Ibrahim, Synthesis, characterization, and
antimicrobial properties of copper nanoparticles, Int. J. Nanomed. 8 (2013) 44674479.
[5] Q. Chen, Y. Xue, J. Sun, Kupfer cell-mediated hepatic injury induced by silica nanoparticles in vitro and in vivo,
Int. J. Nanomed. 8 (2013) 11291140.
[6] S. Pal, Y.K. Tak, J.M. Song, Does the antibacterial activity of silver nanoparticles depend on the shape of the nanoparticle a
study of the gram-negative bacterium Escherichia coli, Appl. Environ. Microbiol. 73 (2007) 17121720.
[7] M. Zarei, A. Jamnejad, E. Khajehali, Antibacterial effect of silver nanoparticles against four foodborne pathogens,
Jundishapur J. Microbiol. 7 (2014) E8720.
[8] G.P. Tegos, T.N. Demidova, D. Arcila-Lopez, H. Lee, T. Wharton, H. Gali, Cationic fullerenes are effective and selective
antimicrobial photosensitizers, Chem. Biol. 12 (2005) 11271135.
[9] C. Yang, J. Mamouni, Y. Tang, L. Yang, Antimicrobial activity of single-walled carbon nanotubes, length effect, Langmuir 26
(2010) 1601316019.
[10] K.S. Novoselov, A.K. Geim, S.V. Morozov, D. Jiang, Y. Zhang, S.V. Dubonos, I.V. Grigorieva, A.A. Firsov, Electric eld effect in
atomically thin carbon lms, Science 306 (2004) 666669.
[11] S. Latil, L. Henrard, Charge carriers in few-layer graphene lms, Phys. Rev. Lett. 97 (2006) 036803-1036803-4.
[12] A.A. Balandin, S. Ghosh, W.Z. Bao, I. Calizo, D. Teweldebrhan, F. Miao, C.N. Lau, Superior thermal conductivity of singlelayer graphene, Nano Lett. 8 (2008) 902907.
[13] L.L. Zhang, S.Y. Zhao, X.N. Tian, X.S. Zhao, Layered graphene oxide nanostructures with sandwiched conducting polymers as
supercapacitor electrodes, Langmuir 26 (2010) 1762417628.
[14] Y. Xuan, Y.Q. Wu, T. Shen, M. Qi, M.A. Capano, J.A. Cooper, P.D. Ye, Atomic-layer-deposited nanostructures for graphenebased nanoelectronics, Appl. Phys. Lett. 92 (2008) 013101-1013101-3.
[15] X.M. Yang, Y.F. Tu, L. Li, S. Shang, X.M. Tao, Well-dispersed chitosan/graphene oxide nanocomposites, ACS Appl. Mater.
Interfaces 2 (2010) 17071713.
[16] H. Fan, L. Wang, K. Zhao, N. Li, Z. Shi, Z. Ge, Z. Jin, Fabrication, mechanical properties and biocompatibility of graphene
reinforced chitosan composites, Biomacromolecules 11 (2010) 23452351.
[17] Mohsen Ahmadipour, K. Venkateswara Rao, V. Rajendar, Formation of nano scale Mg(x) Fe (1  X) O(X = 0.1, 0.2, 0.4)
structure by solution combustion: effect of fuel to oxidizer ratio, J. Nanomater. (2012) (8 pages).

784

S. Bykkam et al. / Superlattices and Microstructures 83 (2015) 776784

[18] S.T. Sun, P.Y. Wu, A one-step strategy for thermal- and pH-responsive graphene oxide interpenetrating polymer hy-drogel
networks, J. Mater. Chem. 21 (2011) 40954097.
[19] M. Terrones, O. Martn, M. Gonzlez, J. Pozuelo, B. Serrano, J.C. Cabanelas, S.M. Vega-Daz, J. Baselga, Interphases in
graphene polymer-based nanocomposites: achievements and challenges, Adv. Mater. 23 (2011) 53025310.
[20] H. Kim, A.A. Abdala, C.W. Macosko, Graphene/polymer nanocomposites, Macromolecule-s 43 (2010) 65156530.
[21] T. Kuilla, S. Bhadra, D. Yao, N.H. Kim, S. Bose, J.H. Lee, Recent advances in graphene based polymer composites, Prog. Polym.
Sci. 35 (2010) 13501375.
[22] C. Liu, S. Alwarappan, Z.F. Chen, X.X. Kong, C.Z. Li, Membraneless enzymatic biofuel cells based on graphene nanosheets,
Biosens. Bioelectron. 25 (2010) 18291833.
[23] M.D. Stoller, S.J. Park, Y.W. Zhu, J.H. An, R.S. Ruoff, Graphene-based ultracapacitors, Nano Lett. 8 (2008) 34983502.
[24] L. Wang, K. Lee, Y.Y. Sun, M. Lucking, Z. Chen, J.J. Zhao, S.B. Zhang, Graphene oxide as an ideal substrate for hydrogen
storage, ACS Nano 3 (2009) 29953000.
[25] E. Yoo, J. Kim, E. Hosono, H.S. Zhou, T. Kudo, I. Honma, Large reversible Li storage of graphene nanosheet families for use in
rechargeable lithium ion batteries, Nano Lett. 8 (2008) 22772282.
[26] X. Wang, L. Zhi, K. Mullen, Transparent, conductive graphene electrodes for dye-sensitized solar cells, Nanoletters 8 (2008)
323327.
[27] X. Li, X. Wang, L. Zhang, S. Lee, H. Dai, Chemically derived, ultrasmooth graphene nanoribbon semiconductors, Science 319
(2008) 12291232.
[28] J. Bai, X. Duan, Y. Huang, Rational fabrication of graphene nanoribbons using a nanowire etch mask, Nanoletters 9 (2009)
20832087.
[29] C.C. Lu, Y.C. Lin, C.H. Yeh, J.C. Huang, P.W. Chiu, High mobility exible graphene eld-effect transistors with self-healing
gate dielectrics, ACS Nano 6 (2012) 44694474.
[30] C.H. Lu, H.H. Yang, C.L. Zhu, X. Chen, G.N. Chen, A graphene platform for sensing biomolecules, Angew. Chem. Int. Ed. 48
(2009) 47854787.
[31] L.T. Qu, Y. Liu, J.B. Baek, L. Dai, Nitrogen-doped graphene as efcient metal-free electrocatalyst for oxygen reduction in fuel
cells, ACS Nano 4 (2010) 13211326.
[32] Z. Liu, J.T. Robinson, X.M. Sun, H.J. Dai, PEGylated nanographene oxide for delivery of water-insoluble cancer drug, J. Am.
Chem. Soc. 130 (2008) 1087610877.
[33] X. Sun, Z. Liu, K. Welsher, J. Robinson, A. Goodwin, S. Zaric, H. Dai, Nanographene oxide for cellular imaging and drug
delivery, Nano Res. 1 (2008) 203212.
[34] R. Naresh Kumar, P. Shaikshavali, Vadali V.S.S. Srikanth, K. BhanuSankara Rao, Reducing Agent Free Synthesis of Graphene
from Graphene Oxide, in: Carbon Materials 2012 (CCM12) AIP Conf. Proc. 1538, 2013, pp. 262265.
[35] L. Zhang, Y. Jiang, Y. Ding, M. Povey, D. York, Investigation into the antibacterial behaviour of suspension of ZnO
nanoparticles (ZnO nanouids), J. Nanopart. Res. 9 (2007) 479489.
[36] K.M. Kumar, B.K. Mandal, E.A. Naidu, M. Sinha, K.S. Kumar, P.S. Reddy, Synthesis and characterization of ower shaped zinc
oxide nanostructures and its antimicrobial activity, Spectrochim. Acta, Part A Mol. Biomol. Spectrosc. 104 (2013) 171174.
[37] A. Lipovsky, Y. Nitzan, A. Gedanken, R. Lubart, Antifungal activity of ZnO nanoparticles the role of ROS mediated cell injury,
Nanotechnology 22 (2011) 105101.
[38] S. Vlad, C. Tanase, D. Macocinschi, C. Ciobanu, T. Balaes, D. Filip, I. Gostin, N. Gradinaru, Antifungal behaviour of
polyurethane membranes with zinc oxide nanoparticles, Dig. J. Nanomater, Biostruct. 7 (2012) 5158.
[39] J. You, Y. Zhang, Z. Hu, Bacteria and bacteriophage inactivation by silver and zinc oxide nanoparticles, Colloids Surf. B 85
(2011) 161167.
[40] M. Premanathan, K. Karthikeyan, K. Jeyasubramanian, G. Manivannan, Selective toxicity of ZnO nanoparticles toward
Gram-positive bacteria and cancer cells by apoptosis through lipid peroxidation, Nanomed.: Nanotechnol., Biol. Med. 7
(2012) 184192.
[41] A. Kahru, H.-C. Dubourguier, I. Blinova, A. Ivask, K. Kasemets, Biotests and biosensors for ecotoxicology of metal oxide
nanoparticles, a Minireview, Sensors 8 (2008) 51535170.
[42] R. Brayner, R. Ferrari-Iliou, N. Brivois, S. Djediat, M-F. Benedetti, F. Fievet, Toxicological impact studies based on Escherichia
coli bacteria in ultrane ZnO nanoparticles colloidal medium, Nano Lett. 6 (2006) 866870.
[43] Z. Huang, X. Zheng, D. Yan, G. Yin, X. Liao, Y. Kang, Y. Yao, D. Huang, B. Hao, Toxicological effect of ZnO nanoparticles based
on bacteria, Langmuir 24 (2008) 41404144.
[44] Y. Li, W. Zhang, J. Niu, Y. Chen, Mechanism of photogenerated reactive oxygen species and correlation with the
antibacterial properties of engineered metal-oxide nanoparticles, ACS Nano 6 (2012) 51645173.
[45] F. Tuinstra, J.L. Koenig, Raman spectrum of graphite, J. Chem. Phys. 53 (1970) 11261130.
[46] A.C. Ferrari, J. Robertson, Interpretation of Raman spectra of disordered and amorphous carbon, Phys. Rev. B 61 (2000)
1409514107.
[47] D. Graf, F. Molitor, K. Ensslin, C. Stampfer, A. Jungen, C. Hierold, Spatially resolved Raman spectroscopy of single and few
layer graphene, Nano Lett. 7 (2007) 238242.