A simple one-spot synthetic route for the production of Ag-nanoparticles using aqueous extract of citrus lemon is being reported in presence of shape-directing cetyltrimethylammonium bromide (CTAB). To our knowledge, this is the first report where the biomolecules form a layer around a group of the Ag-nanoparticles in which the inner layer is bound to the AgNPs surface via the hydroxyl groups of citric acid. The appearance of a sharp surface plasmon resonance band in the UV-visible region might be due to the formation of spherical Ag-nanoparticles. Agglomeration number (N Ag), the average number of silver atoms per nanoparticle (N), molar concentrations of nanoparticle (C) in solution, extinction coefficient (ε) and increase in the Fermi energy (ΔE F) were calculated using Mie theory and discussed. Interestingly, reaction mixture became turbid at higher [CTAB] due to the uncontrolled growth of Ag-nanoparticles. The transmission electron microscopic images of nanoparticles, recorded at different magnifications.