Fatima mohammed Abedalwahab An-Najah National University Antibacterial and Antioxidant Activity of Ephedra alata Decne (alanda) plant Fatima mohammed Abedalwahab Lecture Pharmacy department An-Najah National University

Research objectives Isolation of different extracts from Ephedra alata Decne (alanda) Qualitative and quantitative Phytochemical screening for active ingredients in these extracts. Investigating the antioxidant activity of this extract using DPPH free radical scavenging assay. Investigating the antibacterial activity of this plant against different strains of microorganisms.

alanda plant description Ephedra alata Decne (Latin scientific name) Alanda (Arabic common name) Family: Ephedraceae The native land for this species is Algeria, Palestine, Lebanon, Jordan, Saudi Arabia, Morocco. perennial genus of non flowering seed herb with light green densely branched dioecious small and perennial stiff shrub, about 50-100cm

Alanda plant in the folk medicine Stimulant as energy drink Treatment for UTI problems Relief asthma attack Digestive system disorders Chewing plant stems for treatment of bacterial and fungal infections

Experimental examination for phytochemicals In alanda plant

Soxhlet extraction

Qualitative Phytochemical Tests compounds Aqueous extract Methanol extract Acetone extract Ethanol extract Cardiac glycosides +++ - + Saponin glycoside Alkaloids ++++ Amino acids Starch Reducing sugars Phenols ++ Volatile oil Tannin Steroids Flavonoid

Quantitative Total phenol &Total flavonoid Determination Gallic acid as reference for TPC Gallic acid concentration (mg/ml) Absorbance (mean value) at λmax =765nm 0.0312 0.241 0.0468 0.348 0.0625 0.393 0.125 0.922 0.25 2.021

Rutin as reference for TFC Rutin concentration (μg/ml) Absorbance (mean value) at λmax =415 0.3 0.143 0.6 0.197 1.2 0.213 2.4 0.344 6 0.563

Quantitative Total phenol & Total flavonoid Determination E. alata extract Total phenolic content (mg GA/g) ±SD Total flavonoid content (mg RU/g) ±SD Aqueous extract --- 0.519 ±0.09 Methanol extract 47.62 ±0.94 54.66 ±0.12 Ethanol extract 19.175 ±0.625 5.44 ±0.625 Total phenolic and total flavonoid contents of different Ephedra alata extracts

Experimental examination for antioxidant activity Of alanda plant

DPPH scavenging assay After reduction of Free radical by alata

% inhibition by Trolox ±SD % inhibition by Ephedra alata ±SD Result for antioxidant activity Conc µg/ml % inhibition by Trolox ±SD % inhibition by Ephedra alata ±SD 1 38.65 ±1.08 29.14 ±1.11 2 47.55 ±1.34 29.38 ±1.72 3 52.09 ±1.05 32.09 ±1.28 5 64.19 ±1.32 32.09 ±0.96 7 64.19 ±1.83 33.33 ±1.21 10 69.38 ±1.33 38.76 ±1.53 20 76.29 ±2.12 43.46 ±1.65 30 81.23 ±1.43 46.17 ±1.77 40 92.1 ±1.65 71.36 ±1.21 50 96.05 ±1.40 73.83 ±1.43 80 98.02 ±1.87 73.83 ±1.11 100 98.32 ±1.58 75.02 ±1.67 IC50= 16.03 µg/ml for Ephedra alata IC50= 3.6 µg/ml for Trolox standard.

Antibacterial activity evaluation

Result after micro dilution method MIC (mg/ml) Bacterial Isolates Nonpolar column Polar extract Nonpolar extract PC3 PC2 PC1 41 250 4.1 54.5 2.5 15 S. aureus (ATCC 25923) No inhibition 218 0.32 960 E. coli (ATCC 25922) 125 109 5.1 120 P. aeruginosa (ATCC 27853) 240 MRSA

Conclusion The Phytochemical screening showed that the Ephedra alata plant extract contain a mixture of phytochemicals as cardiac glycosides, reducing sugars, flavonoid, phenolic compounds and alkaloids. The quantitative total flavonoids and total phenol screening indicated that the methanolic plant extract has the highest contents of flavonoids and phenols DPPH assay showed that the plant has potent antioxidant activity which can be further subjected for the isolation of the therapeutically active compounds with anticancer potency. The aqueous extract showed potent antibacterial effect comparing to organic extract especially E.coli bacterial strain