Accepted Manuscript

Synthesis, Characteristics and Antimicrobial activity of ZnO nanoparticles

A. Chinnammal Janaki, E. Sailatha, S. Gunasekaran

PII: S1386-1425(15)00193-6
DOI: [Link]
Reference: SAA 13331

To appear in: Spectrochimica Acta Part A: Molecular and Biomo-

Received Date: 23 October 2014

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: [Link]

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Synthesis, Characteristics and Antimicrobial activity of ZnO nanoparticles

A. Chinnammal Janakia, E. Sailathaa, S. Gunasekaranb

*Corresponding Author E-mail: rsundara3 @[Link]

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 bio-

components 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 anti-

inflammatory 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 UV-

B, 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.
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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
5

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

[Link]. [15] and Y. [Link]. [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.
-1
Metal oxides generally give absorption in fingerprint region, i.e. below 1000 cm arising from

inter- atomic vibrations. It reveals that the vibrational wavenumbers

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

[Link] 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]. [Link] et al. reported the antibacterial
8

activity of ZnO nanoparticles against [Link], [Link], [Link]. 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.

[Link] et. Al has reported the antifungal activity of ZnO NP against [Link], [Link],

[Link] and [Link] [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
9

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

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[10] A. Saxena, R. M. Tripathi, R. P. Singh, Digest Journal of Nanomaterials and Biostructures

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(2000) 36-45.

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Chemistry (2012) 1-9.

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[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, [Link], [Link], [Link], [Link], C. Zhong, B. Wallek, and S. Wong, [Link],
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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.

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Medicine 19 (2008) 1407-1412.

Fig. 1 XRD pattern of synthesized ZnO nanoparticles using dry ginger rhizome
13

Fig. 2 SEM images of ZnO nanoparticles

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

35 865

30

25 2926
1192
20

15

10 1495 1105 549

0.0
4000.0 3600 3200 2800 2400 2000 1800 1600 1400 1200 1000 800 600 450.0
cm-1

Fig. 4 FTIR spectrum of ZnO nanoparticle

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

Fig. 8 Inhibition zone of ZnO nanoparticles against Penicillium notatum

Zone of Inhibition in mm
Streptomycin
[Link]. Micro organisms 1000 μg 500 μg 250 μg 125 μg 62.5 μg DMSO 10 μg
Nano
Klebsiella
1 pneumonia 11 10 9 9 9 - 16

Table 2

Zone of Inhibition in mm
Streptomycin
[Link]. Micro organisms 1000 μg 500 μg 250 μg 125 μg 62.5 μg DMSO 10 μg
Nano
Staphylococcus
1 aureus 10 9 9 9 9 - 20

Table 3

Zone of Inhibition in mm
Streptomycin
[Link]. Micro organisms 1000 μg 500 μg 250 μg 125 μg 62.5 μg DMSO 10 μg
Nano
1 C. albicans 10 9 9 8 7 - 7

Table 4

Zone of Inhibition in mm
Streptomycin
[Link]. Micro organisms 1000 μg 500 μg 250 μg 125 μg 62.5 μg DMSO 10 μg
Nano
Penicillium
1 notatum 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.