Rajiv, 2013. Bio-Fabrication of Zinc Oxide Nanoparticles Using Leaf Extract of Parthenium Hysterophorus L. and Its Size-Dependent Antifungal Activity Against Plant Fungal Pathogens
Rajiv, 2013. Bio-Fabrication of Zinc Oxide Nanoparticles Using Leaf Extract of Parthenium Hysterophorus L. and Its Size-Dependent Antifungal Activity Against Plant Fungal Pathogens
Rajiv, 2013. Bio-Fabrication of Zinc Oxide Nanoparticles Using Leaf Extract of Parthenium Hysterophorus L. and Its Size-Dependent Antifungal Activity Against Plant Fungal Pathogens
h i g h l i g h t s g r a p h i c a l a b s t r a c t
a r t i c l e i n f o a b s t r a c t
Article history: The study reports the synthesis and characterization of zinc oxide nanoparticles from weed plant by a
Received 16 October 2012 novel method. The aim of this work is to synthesize zinc oxide nanoparticles from Parthenium hysteropho-
Received in revised form 10 April 2013 rus L. by inexpensive, ecofriendly and simple method. Highly stable, spherical and hexagonal zinc oxide
Accepted 15 April 2013
nanoparticles were synthesized by using different concentrations of 50% and 25% parthenium leaf
Available online 25 April 2013
extracts. Both the concentrations of the leaf extract act as reducing and capping agent for conversion of
nanoparticles. Formation of zinc oxide nanoparticles have been conrmed by UVVis absorption spectros-
Keywords:
copy, X-ray diffraction (XRD), Fourier trans-form infrared spectroscopy (FT-IR), scanning electron micros-
Parthenium
Antifungal activity
copy (SEM) and transmission electron microscopy (TEM) analysis with energy dispersive X-ray analysis
SEM and TEM (EDX). SEM, TEM and EDX analysis reveals that spherical and hexagonal zinc oxide nanoparticle sizes were
Zinc oxide nanoparticles 27 5 nm and 84 2 nm respectively and chemical composition of zinc oxide were present. We synthe-
Size-dependent sized different sized zinc oxide nanoparticles and explored the size-dependent antifungal activity against
plant fungal pathogens. Highest zone of inhibition was observed in 25 lg/ml of 27 5 nm size zinc oxide
nanoparticles against Aspergillus avus and Aspergillus niger. Parthenium mediated zinc oxide nanoparticles
were synthesized and proved to be good antifungal agents and environment friendly.
2013 Published by Elsevier B.V.
Introduction materials because of their low melting point, special optical prop-
erties, high catalytic activity, and unusual mechanical properties
Particles smaller than tens of nanometers in primary particle compared with their bulk material counterpart [1]. Zinc oxide is
diameter (nanoparticles) are of interest for the synthesis of new attracting tremendous attention due to its interesting properties
Corresponding author. Tel./fax: +91 4222611146. like wide direct band gap of 3.3 eV at room temperature and high
E-mail address: [email protected] (S. Rajeshwari). exciton binding energy of 60 meV [2]. Zinc oxide is a unique
material that exhibits semiconducting, piezoelectric, and pyroelec- carefully and nely cut. Samples were ground well by mortar
tric properties and has versatile applications in transparent elec- and pistle using de-ionized water. The mixture of plant extract
tronics, ultraviolet (UV) light emitters, piezoelectric devices, was heated with medium ame. After cooling this solution was l-
chemical sensors, spin electronics, personal care products, and tered using lter paper (Whatman No. 42, Maidstone, England) and
coating and paints [35]. In fact, zinc oxide is non-toxic and chem- store in refrigerator for further studies.
ically stable under exposure to both high temperatures and UV [6]. Zinc nitrate was dissolved in de-ionized water under constant
Plants and/or their extracts provide a biological synthesis route stirring. The 50% and 25% of plant extract was prepared with de-
of several metallic nanoparticles which are more eco-friendly and ionized water and the volume was made up to 250 ml. In order
allows a controlled synthesis with well-dened size and shape to, the zinc nitrate solution was added in plant extract under con-
[7]. Green synthesis of gold nanoparticles using plants such as stant stirring. This mixture of the solution was kept under vigorous
neem [8], lemon grass [9], alfalfa [10], Emblica ofcinalis [11], and stirring at 90 C for 45 h. After this process a green color precipi-
Cinnamomum camphora [12] have been reported. The use of envi- tate was obtained. This mixture was centrifuged at 7000 rpm for
ronmentally benign materials like plant leaf extract [13], bacteria 15 min and the green precipitate was discarded. The supernatant
[14], fungi [15] and enzymes [16] for the synthesis of silver nano- was stirred again at 150 C for 1 h and a pale yellow color solid pre-
particles offers numerous benets of eco-friendliness and compat- cipitate was obtained. The precipitate was washed with methanol
ibility for pharmaceutical and other biomedical applications as and air dried. This product was annealed at 400 C for 1 h. Finally,
they do not use toxic chemicals for the synthesis protocol. The en- white color powder was obtained. The whole experiment was re-
zymes [17], plant leaf extract [18] and bacteria [19] were vital role peated three times. The methodology is given in the Supplemen-
in green synthesis of zinc oxide nanoparticles. tary information A1.
Plants are often affected by plant pathogens such as fungi, bac-
teria and viruses which results in great agricultural loss [20]. Sev- Characterization of zinc oxide nanoparticles
eral conventional methods have been used for the control or
reduce of these pathogens and each of these methods has one or The aqueous zinc oxide nanoparticles were characterized by UV
more limitations (pesticides cause dangerous effect on the envi- absorption spectra (UV-2450, shimadzu). The synthesized zinc
ronment and human health). Thus, use of nanoparticles has been oxide nanoparticles purity and grain size were identied by X-
considered as an alternate and effective approach which is eco- ray diffraction (PerkinElmer spectrum one instrument) Cu Ka
friendly and inexpensive for the control of pathogenic microbes radiations (k = 0.15406 nm) in 2h range from 20 to 80. The pow-
[21]. Nanoparticles have a vital role in the management of plant der sample of zinc oxide nanoparticles functional groups were ana-
diseases as compared to synthetic fungicides [22]. Zinc oxide nano- lyzed by using Fourier trans-form infrared spectroscopy (Perkin
particles have also been proposed as an anti-microbial preservative Elmer 1725). The nanoparticles size and shape were character-
for wood or food products [2325]. The size dependent activity ized by using scanning electron microscopy (Model JSM 6390LV,
was studied in the range of 100 nm to 0.8 lm in Staphylococcus JOEL, USA). The nanoparticles average size and size distribution
aureus and Escherichia coli and 12, 45, and 200 nm range in were determined by transmission electron microscopy (JEOL
E. coli, however little is known about the activity of zinc oxide JEM-3100F). The samples analyzed for energy dispersive X-ray
nanoparticles in the 10300 nm range toward S. aureus [26,27]. analysis (RONTECs EDX system, Model QuanTax 200, Germany).
Parthenium hysterophorus L. (family Asteraceae) is one of the 10
worst weeds in the world [28], which is a poisonous, pernicious Determination of size-dependent antifungal activity of zinc oxide
and aggressive weed and is reported to have pharmacological nanoparticles by the well-diffusion method
properties against many diseases such as rheumatism, hepatic
amoebiasis and tumours [2931]. Parthenium is also known to Antifungal activities of the synthesized zinc oxide nanoparticles
cause allergy and toxicity in animals [32]. In this study, we synthe- of different sizes were determined using plant fungal pathogens by
sized two zinc oxide nanoparticles of varying sizes and tested their a modied Kirby Bauer disc diffusion method [33]. The fungi were
antifungal activity using well diffusion method. cultured in potato dextrose broth at room temperature on an orbital
shaking incubator (Remi, India) at 200 rpm. A 100 lL of broth fun-
gal culture was prepared and spread on potato dextrose agar plates.
Materials and methods After that plates were allowed to stand for 10 min to allow for cul-
ture absorption. The 5 mm size wells were punched into the agar
Materials with help of sterile gel puncher. A 100 lL (25 lg/ml) of the nano-
particles solution sample and 100 lL (10 lg/ml) of positive control
P. hysterophorus L. plants were collected from follow lands in (amphotricin B) were poured into wells on all plates using micropi-
and around Karpagam University, Eachanari, Coimbatore, India pette. After incubation at room temperature for 48 h, the size of the
(11160 N; 76580 E) on 2011 before owering stage. All the chemi- zone of inhibition diameter in millimeter was measured. Each
cals were obtained from sigma-aldrich chemicals, India. Aspergillus screening treatment was conducted with three replicates and the
avus (MTCC-7589), Aspergillus niger (MTCC-2587), Aspergillus results are presented as mean SE (standard error of the mean).
fumigatus (MTCC-2550) Fusarium culmorum (MTCC-2090) and
Fusarium oxysporium (MTCC-3379) were obtained from the Depart-
Results and discussion
ment of Microbiology, School of Life Sciences, Karpagam Univer-
sity, Coimbatore, India. The culture samples were maintained on
Characterization of zinc oxide nanoparticles
potato dextrose broth at room temperature (aerobically) at
200 rpm shaking for 3 days.
The UVVisible absorption spectra of the as-grown a new tech-
nique for the preparation of the monodispersed semiconductor
Synthesis of zinc oxide nanoparticles using Parthenium leaf extract zinc oxide nanoparticles (50% and 25% plant extract) are shown
in Fig. 1a and b. The room temperature spectra exhibit strong exci-
Zinc nitrate was used as a precursor material for nanoparticles tonic absorption peaks at 374.00 nm (3.32 eV) and 370.50 nm for
synthesis. The harvested plants were washed with tap water and samples 50% and 25% plant extract, respectively, which is in good
whipped with tissue paper. The plant leaf samples were weighed agreement with the previous work [19,34,35].
386 P. Rajiv et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 112 (2013) 384387
Fig. 1. UVvis spectroscopy of synthesized zinc oxide nanoparticles. (A) 25% Extract
and (B) 50% extract. Fig. 3a. Analysis of chemical composition of zinc oxide nanoparticles (25% extract).
P. Rajiv et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 112 (2013) 384387 387