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Nanomaterials 2020, 10, 422 3 of 16 suitable volume of suspension was added to 250 mL Erlenmeyer flasks containing 100 mL of broth to yield an initial concentration of 106/mL blastospores. Figure 1. Synthesis of AgNPs from the leaf extract of Calotropis gigantean (CG). (A) Candida albicans isolated from sputum samples, cultured on Sabouraud’s dextrose agar (SDA). (B) Silver nitrate solution before (A) and after (B) the synthesis of AgNPs from the leaf extract of C. gigantean. 2.4. Preparation and Characterization of AgNPs 2.4.1. Preparation of the Leaf Extract Fresh leaves were washed with distilled water, then cut into small pieces and allowed to dry at room temperature. Ten grams of leaves were boiled in 100 mL distilled water for 20 min and filtered through Whatman No. 1 filter paper [20]. 2.4.2. Biosynthesis of AgNPs An aqueous solution (1 mM) of silver nitrate (AgNO3) was mixed with leaf extract. The mixture was kept in a microwave oven at 300 W for 4 min to complete the bio-reduction of AgNO3 to Ag+ ions. Complete reduction was confirmed by changing the color from colorless to colloidal brown and saturation was detected using UV-visible spectrophotometric scanning with an Agilent 8453 spectrophotometer (Santa Clara, CA, USA) [20]. 2.4.3. Transmission Electron Microscopy (TEM) TEM was performed according to the method of Teranishi et al. [21] on a Leo 912 AB instrument (Aalen, Germany). Briefly, a drop of diluted sample of AgNPs was poured onto carbon-coated copper grids and allowed to stand for 2 min before imaging. 2.4.4. X-ray Diffraction Analysis (XRD) The lyophilized AgNPs coated on an XRD grid were exposed to XRD measurements. The analysis was carried out in an X-ray diffractometer with an operating voltage of 45 kV and a current of 0.8 mAPDF Image | Inhibition of Candidiasis Calotropis Silver Nanoparticles
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