5) for 1 h at 37 °C and the cleavage of caspase-3 substrate was measured at an excitation wavelength of 390 nm and an emission wavelength of 460 nm. The activity was expressed as relative fluorescence unit (RFU). To investigate the internucleosomal DNA fragmentation caused by both silver and gold nanoparticles, DNA laddering assay was performed according to the standard procedure described by Su et al. (2005) with little modification [38]. A total of 1 × 106 cells was treated with silver and gold nanoparticles (100 μg/ml) Selleck H 89 for 48 h and then collected by centrifugation. Further, the DNA was isolated using commercially available

kit (Genei, Bangalore, India) following the manufacturer’s instructions. DNA was resolved on 1.5% agarose gel (containing 3 μg/ml of ethidium bromide in 1 × TAE buffer of pH 8.5) at 90 V for 1.5 h and the bands were visualized using UV transilluminator. In this present study, gold nanoparticles were rapidly synthesized using A. indica leaves extract as bio-reductants. Similar to silver nanoparticles formation, the bio-reduction of HAuCl4 into gold nanoparticles was completed within 30 min of

incubation. The very first indication for nanoparticles formation is colour change. A clear pinkish violet colour was formed within 30 min when 1 mM Alectinib HAuCl4 was added into the aqueous leaves extract of A. indica, which indicates the biogenic synthesis of gold nanoparticles ( Fig. 1). The intensity of pinkish violet colour was increased with the incubation period and it was due

to the excitation of Etomidate surface plasmon vibrations. On the other hand, control (leaf extract alone) showed no change of colour ( Fig. 1). Very recently, Karuppaiya et al. (2013) have reported that the aqueous extract of Dysosma pleiantha rhizome rapidly biosynthesized gold nanoparticles within 20 min [25]. A characteristic absorption peak at 540 nm further confirmed the formation of nano-sized gold particles ( Fig. 2). The formation of gold nanoparticles was started at 15 min and was completed at 30 min. Interestingly, the peak was found to be stable at the same wave length for up to 1 h, indicating that phytochemicals may have stabilized the synthesized gold nanoparticles ( Fig. 2). Fig. 3a and b depict digitalized FE–SEM and TEM images of biosynthesized gold nanoparticles, respectively. These two images showed spherical shaped gold nanoparticles with a size of less than 30 nm. XRD analysis showed three distinct diffraction peaks at 38.1°, 44.1° and 64.1° which indexed the planes 1 1 1, 2 0 0 and 2 2 0 of the cubic face-centred gold. The obtained data was matched well with the Joint Committee on Powder Diffraction Standards (JCPDS) file no. 04–0784, which suggest the crystalline nature of gold nanoparticles ( Fig. 4).