Determination of Antioxidant Activity by Oxygen Radical Absorbance Capacity (ORAC-FL), Cellular Antioxidant Activity (CAA), Electrochemical and Microbiological Analyses of Silver Nanoparticles Using the Aqueous Leaf Extract of Solanum mammosum L.

Abstract

Fernanda Pilaquinga,1,2 Jeroni Morey,2 Lenys Fernandez,1 Patricio Espinoza-Montero,1 Mauricio Moncada-Basualto,3 Josue Pozo-Martinez,3 Claudio Olea-Azar,3 Rafael Bosch,4 Lorena Meneses,1 Alexis Debut,5 María de las Nieves Piña2 1School of Chemical Sciences, Pontificia Universidad Católica del Ecuador, Quito, Ecuador; 2Department of Chemistry, University of the Balearic Islands, Palma de Mallorca, Spain; 3Department of Inorganic and Analytical Chemistry, University of Chile, Santiago, Chile; 4Environmental Microbiology, IMEDEA (CSIC-UIB), and Microbiology, Department of Biology, University of Balearic Islands, Palma de Mallorca, Spain; 5Centro de Nanociencia y Nanotecnología, Universidad de las Fuerzas Armadas ESPE, Sangolquí, EcuadorCorrespondence: Fernanda PilaquingaSchool of Chemical Sciences, Pontificia Universidad Católica del Ecuador, Quito, EcuadorTel [phone]Email [email protected]: The importance of studying polyphenolic compounds as natural antioxidants has encouraged the search for new methods of analysis that are quick and simple. The synthesis of silver nanoparticles (AgNPs) using plant extracts has been presented as an alternative to determine the total polyphenolic content and its antioxidant activity.Methods: In this study, aqueous leaf extract of Solanum mammosum, a species of plant endemic to South America, was used to produce AgNPs. The technique of oxygen radical absorption capacity using fluorescein (ORAC-FL) was used to measure antioxidant activity. The oxidation of the 2´,7´-dichlorodihydrofluorescein diacetate (DCFH2-DA) as fluorescent probe was used to measure cellular antioxidant activity (CAA). Electrochemical behavior was also examined using differential pulse voltammetry (DPV) and cyclic voltammetry (CV). Total polyphenolic content (TPH) was analyzed using the Folin-Ciocalteu method, and the major polyphenolic compound was analyzed by high performance liquid chromatography with diode array detector (HPLC/DAD). Finally, a microbial analysis was conducted using Escherichia coli and Bacillus sp.Results: The average size of nanoparticles was 5.2 ± 2.3 nm measured by high-resolution transmission electron microscopy (HR-TEM). The antioxidant activity measured by ORAC-FL in the extract and nanoparticles were 3944 ± 112 and 637.5 ± 14.8 μM ET/g of sample, respectively. Cellular antioxidant activity was 14.7 ± 0.2 for the aqueous extract and 12.5 ± 0.2 for the nanoparticles. The electrochemical index (EI) was 402 μA/V for the extract and 324 μA/V for the nanoparticles. Total polyphenolic content was 826.6 ± 20.9 and 139.7 ± 20.9 mg EGA/100 g of sample. Gallic acid was the main polyphenolic compound present in the leaf extract. Microbiological analysis revealed that although leaf extract was not toxic for Escherichia coli and Bacillus sp., minor toxic activity for AgNPs was detected for both strains.Conclusion: It is concluded that the aqueous extract of the leaves of S. mammosum contains nontoxic antioxidant compounds capable of producing AgNPs. The methods using AgNPs can be used as a fast analytical tool to monitor the presence of water-soluble polyphenolic compounds from plant origin. Analysis and detection of new antioxidants from plant extracts may be potentially applicable in biomedicine.Keywords: silver nanoparticles, Solanum mammosum, antioxidant activity, ORAC, voltammetry

Keywords

• silver nanoparticles
• solanum mammosum
• antioxidant activity
• orac
• voltammetry