Accepted Manuscript Synthesis, Characteristics and Antimicrobial activity of ZnO nanoparticles A. Chinnammal Janaki, E. Sailatha, S. Gunasekaran PII: DOI: Reference:

S1386-1425(15)00193-6 http://dx.doi.org/10.1016/j.saa.2015.02.041 SAA 13331

To appear in:

Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy

Received Date: Revised Date: Accepted Date:

23 October 2014 29 January 2015 9 February 2015

Please cite this article as: A. Chinnammal Janaki, E. Sailatha, S. Gunasekaran, Synthesis, Characteristics and Antimicrobial activity of ZnO nanoparticles, Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy (2015), doi: http://dx.doi.org/10.1016/j.saa.2015.02.041

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Synthesis, Characteristics and Antimicrobial activity of ZnO nanoparticles A. Chinnammal Janakia, E. Sailathaa, S. Gunasekaranb a

PG and Research Department of Physics, Pachaiyappa’s College, Chennai 600030, TN, India

b

Research and Development, St. Peter’s Institute of Higher Education and Research, St. Peter’s University, Avadi, Chennai 600054, TN, India *Corresponding Author E-mail: rsundara3 @gmail.com Tel.: +91 9176646629 Abstract The utilization of various plant resources for the bio synthesis of metallic nano particles is called Green technology and it does not utilize any harmful protocols. Present Study focuses on the Green synthesis of ZnO nano particles by Zinc carbonate and utilizing the biocomponents of powder extract of dry ginger rhizome. (Zingiber officinale). The ZnO nano crystallites of average size range of 23-26 nm have been synthesized by rapid, simple and eco friendly method. Zinc Oxide nano particles were characterized by using X-Ray diffraction (XRD), Scanning Electron Microscopy (SEM), Energy Diffusion X-Ray spectroscopy (EDX). FTIR spectra confirmed the adsorption of surfactant molecules at the surface of ZnO nanoparticles and the presence of ZnO bonding. Antimicrobial activity of ZnO nano particles was done by well diffusion method against pathogenic organisms like Klebsiella pneumonia, Staphylococcus aureus and Candida albicans and Penicillium notatum. It is observed that the ZnO synthesized in the process has the efficient antimicrobial activity.

1.

Introduction Recent advances in the field of nanotechnology, particularly the ability to prepare highly

2

ordered particulates of any size and shape led to the development of new biocidal agents. Nano materials are called a “wonder of modern medicine “. It is stated that antibiotics kill perhaps a half dozen different disease-causing microorganisms, but nanomaterials can kill some 650 cells [1]. Inorganic metal nano particles have been of great importance due to their distinctive features such as catalytic, optical, magnetic, electronic, and antimicrobial [2,3] wound healing and antiinflammatory properties [4]. Nano particles exhibit novel properties due to the variations in specific characteristics such as size, distribution and morphology of the particles. Among the metal oxide nanoparticles, zinc oxide is interesting because it has vast applications in various areas such as optical, piezoelectric, magnetic, and gas sensing. Besides these properties, ZnO nanostructure exhibits high catalytic efficiency, strong adsorption and are used frequently in the manufacture of sunscreens [5], ceramics, and rubber processing, wastewater treatment, and as a fungicide [6, 7]. In fact, nZnO usage may overtake nano-titanium dioxide(nTiO2) as it can absorb both UV-A and UV-B radiation while nTiO2 can only block UVB, and therefore offering better protection and improved opaqueness[6]. ZnO has large excitation binding energy (60meV) which allows UV lasing action to occur even at room temperature [8] and ZnO with Oxygen vacancies exhibits an efficient green emission. Several physical and chemical procedures have been used for the synthesis of large quantities of metal nanoparticles in a relatively short period of time. But chemical methods lead to the presence of some toxic chemicals adsorbed on the surface that may have adverse effects in medical applications [9]. Currently, plant - mediated Biological synthesis of nanoparticles is gaining importance due to its simplicity, eco friendliness and extensive antimicrobial activity [10, 11]. Bio synthesis of zinc oxide nanoparticles by plant zingiber officinale has been reported.

3

Antimicrobial activity of metal oxides ZnO, MgO and CaO powders were quantitatively evaluated in culture media [12, 13]. It is considered that the detected active oxygen species generated by these metal oxide particles could be the main mechanism of their antibacterial activity. This study therefore is aimed to synthesis ZnO nanoparticles using bio method, to analyse their characteristics using spectroscopic techniques and to evaluate its antimicrobial activity. The plant ginger belonging to the family of Zingeiberaceae and is a common condiment for various foods and beverages. It has ailing history of medicinal use in India for conditions such as headaches, nausea, rheumatism and colds. The anti-inflammatory properties of ginger have been known and valued for centuries. This contains many phytochemicals such as alkaloids, saponins, tannins, and flavanoids. (It includes Panthothenic acid, Vit B6, Folate, Vit C, Calcium, Iron, Magnesium and Manganese). To the best of our knowledge, biological approach using powder extract of dry ginger rhizome has been used for the first time as a reducing material as well as surface stabilizing agent for the synthesis of spherical ZnO nanoparticles. The structure, phase and morphology of synthesized product were investigated by the standard characterization techniques.

2.

Experimental Methods

2.1

Materials Zinc Carbonate (ZnCo3) and glassware were purchased from Gem Co scientific

suppliers, Chennai. All glassware were washed with sterile distilled water and dried in an oven before use.

4

2.2

Bio Synthesis of ZnO nanoparticles using plant extract Dry ginger is made into fine powder. 15 gm of powder is taken. 60 ml of water is added

to it. It is stirred for an hour in a magnetic stirrer. It is filtered using Whatman® qualitative filter paper, Grade 1. Again 40 ml of water is added, stirred again and then filtered. The extract obtained was about 50 ml. The extract (50ml) is heated by stirrer – heater to 57º c. The extract was light yellow in color. When it is at 57º c, 5 gm of Zinc Carbonate is added. Soon it turned to milky white and is then boiled and stirred for 3 hours until it is reduced to a yellow paste. It was carefully collected in a ceramic crucible and heated in an air heated furnace at 400º c for 2 hrs. A light yellow color powder was obtained and collected for characterization purposes. The material was mashed in a mortar – pestle so as to get a fine nature for characterization.

2.3

Characterization techniques Structural and optical properties of the ZnO nanoparticles were determined by using

Scanning Electron Microscopy (SEM) (SAIF, IIT, Madras), X-ray Diffraction (XRD) (UNIVERSITY OF MADRAS, CHENNAI), Fourier Transform Infra-Red spectroscopy (FTIR) (SAIF, IIT, Madras).

3.

Results and Discussion

3.1

X-ray Diffraction (XRD) Analysis The Phase identification of crystalline structure of the nanoparticles was characterised by

X-Ray powder diffraction. The synthesized sample was used by a CuKal- X Ray Diffractometer

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for confirming the presence of ZnO and analyse the structure. Fig.1 shows XRD diffraction pattern of ZNO nanoparticles. It is found that there exist strong diffraction peak with 2θ values of 31.83⁰, 34.42⁰,36.27⁰, 47.48⁰, 56.52⁰, 62.70⁰, and 66.81⁰ corresponding to the crystal planes of (100), (002), (101), (102) (110), (200), and (201) respectively. All diffraction peaks of sample correspond to the characteristic hexagonal wurtzite structure of zinc oxide nanoparticles (a=0.315nm and c=0.529 nm) [14]. Similar, X-ray diffraction pattern was reported by C. Chenn et.al. [15] and Y. Pung.et.al. [16]. The average particle size of ZnO NPs can be estimated using the Debye-Scherer equation [17], which gives a relationship between peak broadening in XRD and particle size that is demonstrated by following equation. d = kλ / ßcosθ Where d is particle size of the crystal, k is Scherer’s constant (0.9), λ is X-Ray wavelength (0.15406nm), ß is the width of the XRD peak at half height and θ is the Bragg diffraction angle. Using the Scherer equation the average crystalline size of ZnO NPs is found to be 24.5 nm. Diffraction pattern corresponding to impurities are found to be absent. This proves that pure ZnO nanoparticles were synthesized.

3.2

Scanning Electron Microscope (SEM) Analysis A scanning electron microscope is a kind of electron microscope which images a sample

by scanning it using a high energy electron beam. The electrons that interact with the atoms making up the sample, thus producing signals which reveal information about the composition, surface topography and other properties such as electrical conductivity.

Thus the surface

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morphology and size of the silver nanoparticles were analyzed by Scanning Electron Microscope. SEM image had shown individual ZnO nanoparticles as well as number of aggregates. Fig.2 illustrates the particles are predominantly spherical in shape and aggregates into larger particles with no well-defined morphology. The SEM image shows the size of the ZnO nanoparticles ranging from 23-25 nm. This size is better compared to previously reported ZnO nps synthesized by Aloe Vera extract which is 40nm [18].

3.3

Energy Dispersive X- ray (EDX) Analysis of ZnO Nano particles EDX profile shows the chemical analysis of synthesized ZnO nano particles. The ZnO

nano particles were used by Hitachi S-3400N SEM instrument equipped with a Thermo EDX attachment. EDX pattern (Fig.3) shows major emission energy at 1 keV which is the binding energy for Zinc (85.30℅) and 0.5 Kev which is the binding energy for Oxygen (13.51℅), which confirms that ZnO has been correctly identified. The pattern shows no other weak signal indicating the purity of ZnO nanoparticles.

3.4

FTIR Analysis of ZnO nanoparticles FTIR spectroscopy is used to identify the functional groups of the active components

based on the peak value in the region of infra-red region. Fig. 4 shows FTIR spectra of ZnO nanoparticles. FTIR measurements were carried out to identify the potential functional groups of the biomolecules in the dry ginger rhizome responsible for the reduction of ZnO nanoparticles. Metal oxides generally give absorption in fingerprint region, i.e. below 1000 cm inter-

atomic

vibrations.

It

reveals

that

the

vibrational

-1

arising from

wavenumbers

7

865,1105,1192,1495,2926,3445 and 3836 cm-1 , correspond to CH, C-OH,CH2- OCH3/CH2-CH3 and OH functional groups present in the alkaloids 6-gingerol,6-Shogal,α – Zingeberene of zingiber officinale are responsible for the reduction of ZnO nanoparticles. From the FTIR result the soluble elements present in ginger extract could have acted as capping agent preventing the aggregate of nano particles in solution and playing a relevant role in their extracellular synthesis and shaping [19]. The peaks at 549 and 621cm-1 are corresponding to ZnO stretching and deformation respectively. The metal oxygen frequencies observed for the respective metal oxide is in accordance with literature values [20, 21].

4.

Antimicrobial Activity Mechanism of action: Antimicrobial activities of metallic NPs are to their high aspect

ratio (size to surface ratio). The nano particles interfere with cellular processes once entering the microbes. Also, the nano particles surface adhesion with the microbial cell surface leads to its immobilization [22]. 4.1

Antibacterial activity of ZnO nanoparticles S.Gunalan et al. reported ZnO, suspension prepared from green synthesis method is more

effective than the suspension with other preparations [18]. This can be explained on the basis of the oxygen species released on the surface of ZnO which cause fatal damage to micro organisms [23]. They react with hydrogen ions to produce molecules of H2O2. The generated H2O2 can penetrate the cell membrane and kill the bacteria. The generation of H2O2 depends strongly on the surface area of ZnO, which results in more oxygen species on the surface and the higher antibacterial activity of the smaller nanoparticles [24]. S.Gunalan et al. reported the antibacterial

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activity of ZnO nanoparticles against S.marcescens, P.mirabilis, C.freundii. Antibacterial activity of ZnO nano sample was determined by disc diffusion method on Muller Hinton Agar (MHA) medium. The synthesized ZnO nanoparticles by green route have more toxic against the bacteria Klebsiella pneumonia and Staphylococcus aureus. The Tables 1 and 2 showed a clear inhibition zone for various concentrations of ZnO nanoparticles. Figs. 5 and 6 showed inhibition zone increased with increasing the concentrations of ZnO nano particles.

4.2

Antifungal activity of ZnO nano particles

Antifungal activity of ZnO was determined by antifungal susceptibility test. The synthesized ZnO nano by green route has more toxic against the fungi Candida albicans and Penicillium notatum. The Tables 3 and 4 showed a clear inhibition zone for various concentrations of ZnO nanoparticles. Figs. 7 and 8 showed the inhibition zone increases with increasing concentrations. S.Gunalan et. Al has reported the antifungal activity of ZnO NP against A.nidulans, T.harzianum, A.flavus and R.stolonifer [18]. To the best of our knowledge for the first time the antifungal activity of ZnO has been reported for Candida albicans and Penicillium notatum.

5.

Conclusions The rapid biological synthesis of ZnO nano particles using a powdered extract of dry

ginger rhizome provides an ecofriendly, simple and efficient route for synthesis of nano particles. The synthesized nano crystallites of ZnO are in the range of 23-26 nm. The morphology of the ZnO particles was characterized by Scanning Electron Microscopy. EDX analysis reveals the

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chemical analysis and proved the purity of ZnO. FTIR analysis confirmed the presence of ZnO bonding. Such eco –friendly nano particles in bactericidal, wound healing and other medical and electronic applications, make this method potentially exciting for the large-scale synthesis of other metal nano particles.

References [1]

T. Sungkaworn, W. Triampo, P. Nalakarn, D. Triampo, I. M. Tang, Y. Lenbury International Journal of Biomedical Science: IJBS2 (2007)67–74.

[2]

A . Ingle, A. Gade, S. Pierrat, C. Sonnichsen, M. Rai, Current Nano science (2008)141– 144.

[3]

N. Duran, P. D. Marcato, O .L. Alves, G. Souza, Journal of Nanotechnology3 (2005)1–7.

[4]

P. L. Taylor, A. L. Ussher, R.E. Burrell, Biomaterials (2005) 7221–7229.

[5]

R. Seshadri, C. N. R. Rao, A. M, A. Muller, A. K. Cheetham (Eds.), The Chemistry of Nanomaterials, Vol.1, Wiley-VCHVerlag GmbH, Weinheim, 2004, 94– 112.

[6]

L. Theodore, Nanotechnology: Basic Calculations for Engineers and Scientists, Wiley, Hoboken, 2006.

[7]

X. Wang, J.Lu, M. Xu, B. Xing, Environmental Science and Technology 42 (2008) 7267–7272.

[8]

Y.K. Park, J. Inhan, M.G. Kwak, H. Yang, S.H. Ju, W.S. Cho, J.Lumin. 78(1998)87.

[9]

D. Jain, H. K. Daima, S. Kachhwaha, S. L . Kothari, Digest Journal of Nanomaterials and Biostructures4 (2009)557–563.

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[10] A. Saxena, R. M. Tripathi, R. P. Singh, Digest Journal of Nanomaterials and Biostructures 5 (2010)427–432. [11] N. Khandelwal, A. Singh, D. Jain, M. K. Upadhyay, H. N. Verma, Digest Journal of Nanomaterials and Biostructures 5 (2010)483–489. [12] J. Sawai Journal of Microbiological Methods54 (2003)177–182. [13] J. Sawai, T. Yoshikawa, Journal of Applied Microbiology 96 (2004)803–809. [14] C. Chen et. al. Joint committee on powder diffraction standards, Diffraction data file, (2000) 36-45. [15] C. Chena. B. Yu, P. Liu, J.F. Liu, L. Wang, Journal of ceramic Processing Research12 (2011) 420-425. [16] Swee-Yong Pung, Wen-pir Lee, Azizan Aziz, International journal of inorganic Chemistry (2012) 1-9. [17] B. D. Culity, Elements of X-Ray diffraction 2nd Ed, Addison-Wesley, USA. [18] G. Sangeetha, S. Rajeswari, R. Venkatesh Progress in Natural Science: Materials International (2012) 693-700. [19] M. Singh, R. Kalaivani, S. Manikandan, N. Sangeetha, A.K. Kumara guru, Appl Nanoscience. 2013,3, 145-151. [20] C. N. R. Rao, Chemical applications of Infrared spectroscopy, Academic Press, Newyork and London, 1963. [21] L. Markova-Deneva, Journal of the University of Chemical Technology and Metallurgy 45 (2010) 351-378. [22] L. Wang, J.Luo, S.Shan, E.Crew, J.yin, C. Zhong, B. Wallek, and S. Wong, Anal.Chem,

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Vol. 83, pp. 8688-8695, 2011. [23] K. Sunanda, y. Kikuchi, K. Hashimoto, A. Fujishima, Environmental Science and Technology 32(1998) 726-728. [24] O.Yamamoto, M. Komatsu, J.Sawai, Z.Nakagwa, Journal of Material Science Materials in Medicine 19 (2008) 1407-1412.

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Fig. 1 XRD pattern of synthesized ZnO nanoparticles using dry ginger rhizome

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Fig. 2 SEM images of ZnO nanoparticles

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Element

Wt%

At%

OK

13.37

38.67

15

ZnK

86.63

61.33

Matrix

Correction

ZAF

Fig. 3 EDX analysis of ZnO nanoparticles

85.6 80 75 604 70 65 60 55

698 3836

2094

621

50 45 %T

40 865

35 30 25

2926 1192

20 15 1495

10

1105

549

3445 5 0.0 4000.0

3600

3200

2800

2400

2000

1800

1600

1400

cm-1

Fig. 4 FTIR spectrum of ZnO nanoparticle

1200

1000

800

600

450.0

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Fig. 5 Inhibition Zone of ZnO Np against Klebsiella pneumonia

Fig. 6 Inhibition Zone of ZnO Np against Staphylococcus aureus

Fig. 7 Inhibition Zone of ZnO Nps against C. albicans

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Fig. 8 Inhibition zone of ZnO nanoparticles against Penicillium notatum

Table 1 Zone of Inhibition in mm S.No.

1

Micro organisms Nano Klebsiella pneumonia

1000 μg

500 μg

250 μg

125 μg

62.5 μg

DMSO

Streptomycin 10 μg

11

10

9

9

9

-

16

Table 2 Zone of Inhibition in mm S.No.

1

Micro organisms Nano Staphylococcus aureus

1000 μg

500 μg

250 μg

125 μg

62.5 μg

DMSO

Streptomycin 10 μg

10

9

9

9

9

-

20

Table 3 Zone of Inhibition in mm S.No.

Micro organisms Nano

1000 μg

500 μg

250 μg

125 μg

62.5 μg

DMSO

Streptomycin 10 μg

1

C. albicans

10

9

9

8

7

-

7

Table 4 Zone of Inhibition in mm S.No.

1

Micro organisms Nano Penicillium notatum

1000 μg

500 μg

250 μg

125 μg

62.5 μg

DMSO

Streptomycin 10 μg

12

12

12

10

8

-

9

18

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Highlights

 The XRD pattern confirmed the crystalline nature of ZnO nanoparticles.  FTIR pattern revealed the presence of ZnO bonding and the functional groups responsible for reduction of ZnO nanoparticles.  Structural morphology of ZnO nanoparticles was studied by SEM analysis.  EDX analysis revealed the chemical composition and purity of synthesised ZnO nano particle.  The ZnO nanoparticles exhibited efficient antimicrobial activity.

Synthesis, characteristics and antimicrobial activity of ZnO nanoparticles.

The utilization of various plant resources for the bio synthesis of metallic nano particles is called green technology and it does not utilize any har...
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