CHAPTER 3: MATERIALS AND METHODOLOGY

3.1 Material required

Chemicals/ Substrate: Fresh Aloe vera leaves, Double distilled water, Silver nitrate, Copper
nitrate Trihydrate.

Apparatus: Test tubes, Pipette, Autoclave facility, Centrifuge machine, Flasks, Beakers,
X-ray diffractometer, UV-Visible Spectrophotometer, Scanning Electron Microscope.

3.2 Methodology

3.2.1 Preparation of leaf extract

Fresh Aloe Vera leaves were collected from a garden and thoroughly washed with double
distilled water to remove dirt and dust on the surface of the leaves. 20g of finely chopped
leaves were added to 100 ml of double distilled water and boiled for 10 minutes. The extract
was cooled down and filtered with Whatman filter paper no. 1 and extract was stored for 4
degree centigrade for further use.

3.2.2 Green Synthesis of silver and copper nanoparticles

100 ml, 1mM solution of silver nitrate and copper nitrate were prepared in separate flasks.
Then 2 ml of plant extract was added to 10 ml of of the metal salt solutions. The two
solutions were autoclaved at 110 degree and 15 psi pressure for 20 minutes. The respective
colour changes in both the solutions were noted to confirm the formation of nanoparticles.

3.2.3 Characterization

1 ml of both the solutions were taken in a cuvette and and subsequently analyzed in a UV
spectrophotometer at room temperature. The spectra was recorded at different wavelengths to
find out the characteristic peak.

The synthesized nanoparticles are centrifuged and then analyzed in electron microscope to
observe the morphology and size.

3.2.4 Antimicrobial Essay

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Antimicrobial potential of silver and copper nanoparticles were tested using agar disk
diffusion method for Lactobacillus. Tests were carried out in absence and then in presence of
visible blue light.

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