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Department of Chemistry, Ch. Devi Lal University, Sirsa (Haryana) 125 055, India
* Corresponding Author: harimoudgil1@rediffmail.com
Abstract- Monoclinic CuO nanoparticles were synthesized by FTIR. Antibacterial activity of CuO nanoparticles were also
microemulsion technique. XRD, TEM and FT-IR techniques investigated against four bacterial strains.
were used to characterize the nanoparticles. Antibacterial activity
of CuO nanoparticles were tested against bacterial strains S. II. EXPERIMENTAL
epidermidis, S. aureus, E. faecalis, and M. luteus using the agar
diffusion method. Minimum Inhibitory Concentration (MIC) of A. Materials and method:
these four bacterial strains S. epidermidis, S. aureus, E. faecalis,
and M. luteus were found to be 0.1, 0.2, 0.6 and 0.8 µg/ml, All chemicals used in experiment were of analytical
respectively. grade. The stable reverse micelle microemulsion was prepared
by mixing a non-ionic surfactant Triton X-100
[(C14H22O(C2H4O)n] (Qualigen Chem. Pvt. Ltd.),Polyvinyl
pyrollidone (PVP) (K85-95) (Merk) and 1:9 ratio of cyclohexane
Keywords- Antibacterial activity, FTIR, TEM, Nanoparticles,
(Qualigen Chem. Pvt. Ltd.) and triple distilled water
XRD.
(conductivity less than 1×10-6 S cm-1) (W/S ratio 5). The
microemulsion was mixed rapidly with continuous stirring for
I. INTRODUCTION five minutes. CuSO4.5H2O (Qualigen Chem. Pvt. Ltd.) solution
(0.5 M) was added drop by drop to microemulsion with
With recent advances in nanotechnology, various types continuous stirring. A sky blue colour mixture was obtained.
of metal and metal oxide nanoparticles with antimicrobial PVP was used as a stabilizing agent. After half an hour of
(microbiocidal or growth-inhibiting) activity have been equilibration, 2.0 M hydrazine hydrate (Qualigen Chem. Pvt.
synthesized [1-9]. Metal nanoparticles containing magnesium Ltd.) solution was added drop by drop with continuous stirring at
oxide [6], copper [7, 8], silver [1-5], iron [10], zinc oxide [11- room temperature. The mixture gradually changed from sky blue
13], and nickel oxide [14, 15] are exhibit antimicrobial to reddish brown without precipitation. The reverse micelles
properties. were broken by adding THF (Merk).
The antimicrobial activity has been observed to vary as a CuO nanoparticles were subsequently washed with
function of surface area in contact with the microbe; therefore ethanol and triple distilled water to remove residual PVP and
nanoparticles with large surface area ensure a broad range of surfactant molecules. After washing CuO nanoparticles were
reactions with the bacterial surface [16]. dried in oven at 100.0 °C for 48 hours.
Compare to other methods, the reverse micelle method is B. Characterization techniques:
one of the most promising wet chemistry synthesis approaches
[17] of synthesis of metal nanoparticles. This method provides a Structural and optical properties of the CuO
favorable microenvironment for controlling the chemical nanoparticles were determined by using Transmission Electron
reaction. As such the reaction rate can be easily controlled, and it Microscopy (TEM) (Hitachi: H-7500; Resolution: 2 Å), X-ray
is possible to obtain a narrow nanoparticle size distribution [18]. Diffraction (XRD) (Rikagu Mini-2 using CuKα1, λ = 0.15406
The size of the core of the reverse micelle can also be controlled nm radiations), and Fourier Transform Infra- Red spectroscopy
by changing water to surfactant ratio [19]. Reverse micelle (FTIR) (Thermo-USA, FTIR-380) in the wavelength range of
microemulsions are transparent, isotropic, and 400 - 4000 cm-1.
thermodynamically stable [20, 21].
(Chandigarh). The culture media; Beef extract and Agar-agar There are two important factors that affect the exchange rate
Type-1 (Hi-Media Pvt. Ltd., Bombay), and chemicals; Peptone, of reverse micelles in microemulsions; the dimer stability and
Sodium Chloride (Qualigen Chemicals Pvt. Ltd., Bombay) were the size of channels between the dimmers [27]. The dimer
used for the growth of bacteria. stability, which depends on the intermicellar attractive potential,
determines the interdroplet transfer of reactants. On the other
D. Antibacterial Activity in Solid Method: hand, the size of channels which depends on the rigidity of
interfacial film in the microemulsion, determines the Ostwald
Antibacterial activity of CuO nanoparticles were tested ripening contribution [28].
against two gram negative bacterial strains of Staphylococcus
epidermidis (MTCC 1809), Staphylococcus aureus (MTCC B. Tranmission Electron Microscopy and X-Ray Diffraction
3160), Enterococcus faecalis (MTCC 2729), and Micrococcus Analysis
luteus (MTCC 1809) using the agar well diffusion assay method
[26]. Approximately, 25.0 ml of molten and cooled nutrient agar Spherical shape of CuO nanoparticles were observed
media were poured in the sterilized petri dishes. The plates were from TEM images (Figure 1a, and b). The average size of copper
left over night at room temperature to check for any oxide nanoparticles was found to be 5.0 – 8.0 nm. It was indicate
contamination to appear. The bacterial test organism S. that surfactant molecules form a film over the surface of CuO
epidermidis, S. aureus, E. faecalis, and M. luteus were grown in nanoparticles which prevented the agglomeration of
nutrient broth for 24 hours at 37 °C. A 100 µl nutrient broth nanoparticles.
culture of each bacterial organism was used to prepare bacterial
lawns. Agar wells were prepared with the help of a sterilized
stainless steel cork borer. Agar wells were prepared with the help
of a sterilized stainless steel cork borer. The wells in each plate
were loaded with 100 µl of different concentrations i.e. 0.10,
0.20, 0.40, 0.60, 0.80, 1.0 and 1.2 µg/ml of copper oxide
nanoparticles.
A. Synthesis
C. FTIR Spectroscopy
D. Antibacterial Activity
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