Characterization of prepared Silver nanoparticles

The bioreduction of pure silver ions was monitored by visual inspection and by measuring the UV-

Vis spectrum of the reaction medium, at the wavelength of 350-500 nm in ELICO (SL-159) UV- Vis

spectrophotometer. Fourier Transform Infrared Spectroscopy (FT-IR) measurements were carried out

to identify the existence of the functional groups in the synthesized AgNPs. The dried and purified

pellets were grounded with KBr and analyzed on an FT-IR instrument. The X-Ray diffraction (XRD)

assay was performed for the detection of crystalline nature of the metal nanoparticles which was done

by powder (X-Ray Diffractometer).

Antibacterial activity

Antibacterial activity of Amphiroa assisted silver nanoparticles was carried out by well diffusion

test technique (Khan et al., 1990) against gram positive and gram negative bacteria. Bacterial

cultures such as Bacillus subtilis, Escherichia coli,

Klebsiella pneumoniae, Staphylococcus aureus, Pseudomonas aeroginosa were acquired from

Bacterial culture collection of SDNB Vaishnav College for Women, Chromepet, Chennai. These

bacterial cultures were freshly cultivated for 24 hrs in nutrient broth. Muller Hinton agar medium was

prepared and poured into sterilized petridishes and allowed to solidify at room temperature. A sterile

cotton swab was used for spreading each test microorganism from the 24 hrs inoculated broth

evenly on the agar plates.

In each of these plates 7 mm diameter wells were made using a sterilized cork borer. Different

concentration of extracts was added to the respective wells on the MH agar plates. Concentration

ranged from 20, 40, 60, 80, and 100

µl, respectively, were placed in the wells and allowed to diffuse at room temperature for 30 min. The

seaweed extracts with no AgNPs was taken as control plate. The AgNPs loaded plates were kept at

37°C for 24 hrs incubation. After incubation, a clear inhibition zone around the wells indicated the

presence of antimicrobial activity. The diameter of the clearing zones was measure in mm using the

ruler scale. The experiment was done in duplicates for each pathogenic bacterium.