Evaluation of In-vitro Pharmacological Activities of Medicinal Plants: Ephedra intermedia (Schrenkex) C.A.Mey and Seriphidium quettense (Podlech) Y.R. Ling Extracts and Their Green Synthesized Silver Nanoparticles

Abstract

Plants-derived drugs has been used for centuries. It is estimated that 80 % of Asian‘s population are defendant of on traditional medicines. In Pakistan about 700 plants species are used for different medicinal purposes. In study Seriphidium quettense (Podlech) and Ephedra intermedia Schrenk & Meye plants were collected from Ziarat district of Balochistan province. Extracts of both plants were obtained using 10 different solvents. Efficiency of each extract was evaluated. Total flavonoids content (TFC), total phenolic content (TPC), total antioxidant activities of these extracts were evaluated. In additional, Seriphidium quettense silver nanoparticles (SqNPs) and Ephedra intermedia silver nanoparticles (EtNPs) were synthesized using aqueous extract of Seriphidium quettense and Ephedra intermedia. Optimization of silver nanoparticles was done using different parameters such a concentration of silver nitrate, pH of reaction mixture, ratio of silver nitrate, plant extract and temperature of reaction mixture. Biogenic silver nanoparticles were characterized by UV spectrophotometer, FT-IR, XRD and SEM. Antimicrobial activity of each extract and their synthesized silver nanoparticles (SqNPs and EtNPs) were evaluated against six different bacterial strains (B. subtilis, E. coli, K. pneumonia, S aureus, P. aeruginosa, and S. epidermidis) and four different fungal strains (A. fumigatus, A. niger, Mucor species and A. flavus). Each solvent extract and Silver nanoparticles were future evaluated for their cytotoxic effects using hemolytic assay and anticancer activity against HepG2 cancer cell lines. We reported maximum percent extract recovery by Ethanol + water (1:1) solvent system. Highest amount of TFC was found in chloroform extracts for both plants. Highest amount of TPC was found in Water + Methanol (1:1) solvent system for both plants. Highest total antioxidant activity was shown by chloroform and methanol + ethanol extract for Seriphidium quettense and Ephedra intermedia respectively. The maximum %FRSA was shown by methanol + ethanol (1:1) extracts for both plants. Nanoparticles synthesized from Seriphidium quettense and Ephedra intermedia showed 45% and 25% %FRSA at 200 µg/ml. The highest reducing power was shown by ethanol extract for both plants. UV–visible spectroscopy results showed that sharp absorbance band was observed at 4mM and 2.5 mM of AgNO3 for SqNPs and EtNPs respectively. The Uv-spectra shows that maximum surface Plasmon Resonance at alkaline pH (pH 8) for SqNPs. The Uv-spectra shows thatMaximum surface Plasmon Resonance when extract to AgNO3 ratio is 1:5 and 1:10 (v/v) for SqNPs and EtNPs respectively. The suitable temperatures for synthesis of SqNPs and EtNPs were found to be 37°C and 30°C respectively. FTIR results of SqNPs and EtNPs show that certain biomolecules are involved in reduction of AgNO3. XRD analysis of biogenic nanoparticles confirm that nanoparticles are of crystalline nature and with average size of 29.16 nm and 29.30 nm SqNPs and EtNPs respectively. SEM results showed that SqNPs and EtNPs are agglomerated and spherical in shape with an average size of 53.3 nm and 49.96 nm respectively. We report that methanolic and methanol + ethanol extracts of Seriphidium quettense showed high antibacterial potential against E. coli and S. aureus respectively. While SqNPs showed high antibacterial activity against B. subtilis, P. aeruginosa, K. pneumonia and S. epidermidis. EtNPs showed high antibacterial activity against B. subtilis, E. coli, P. aeruginosa, K. pneumonia and S. epidermidis. Acetone extract of Ephedra intermedia showed high antibacterial activity against S. aureus. We report that methanol + water extract and chloroform extract of Seriphidium quettense showed high antifungal activity against A. niger, Mucor species and A. flavus. While SqNPs showed high antifungal activity against Aspergillus fumigatus. Chloroform, ethanol, methanol + ethanol and methanol extracts of Ephedra intermedia showed significant antifungal activity against, A. fumigatus, A. flavus, A. niger and Mucor species. Further we report that SqNPs and EtNPs showed anticancer activity as compared to their aqueous extracts against HepG2 liver cell lines. We found that n-hexane and chloroform extract of Seriphidium quettense and ethanol and methanol extracts of Ephedra intermedia showed minimum hemolysis against human erythrocytes. We also found that SqNPs and EtNPs showed acceptable hemolysis at 50 μg/ml concentration.