**2.3 Method for intracellular synthesis of silver nanoparticles**

Prepare the pure bacterial seed culture in 100 ml nutrient broth and incubate it on a rotary shaker (150 rpm) for 24 hours.

*Bacterial Silver Nanoparticles: Method, Mechanism of Synthesis and Application in Mosquito… DOI: http://dx.doi.org/10.5772/intechopen.104144*


#### **2.4 Separation of nanoparticles**

The synthesized nanoparticles are separated from reaction mixture by adapting the following method [12, 15]:

Solution 1: 0.9% NaCl.

Solution 2: NaCl 0.5 M + Sucrose 0.5 M (equal volume of each solution). Solution 3: NaCl [17.5 g/L] + KCL [0.74 g/L] + MgSO4. 7H2O [12.3 g/L] + Tris HCL [0.15 g/L].

pH = 7.5[Salt solution]

#### **Figure 2.**

*Mechanism of intracellular synthesis of silver nanoparticles - bacteria can oxidize metal by an action of reductase enzyme (direct mechanism of gaining electrons from reduced minerals). The redacted Nano metal ions are attached to the surface of the bacterial cell wall.*

