Lakhasly

2.Chemicals
Methanol (MeOH) and dichloromethane were
purchased from Biochem Chemopharma (Montreal,
Quebec, Canada); ethyl acetate, 1-butanol, acetone,
DMSO, 2,2-diphenyl-1-picrylhydrazyl (DPPH), FolinCiocalteu reagent, aluminum chloride (AlCl3), sodium
carbonate (Na2CO3), gallic acid, quercetin, and ferric
chloride (FeCl3) were purchased from Sigma Aldrich,
Chemicals Co (St.The six strains Gramnegative are: Escherichia coli ATCC 25922,
Pseudomonas aeruginosa ATCC 27853, Enterobacter
cloacae, Salmonella enterica, Serratia marcescens and
Vibrio cholerae, and the four strains Gram-positive:
Staphylococcus aureus ATCC 25923, Bacillus subtilis,
Staphylococcus epidermidis and Micrococcus luteus.According to Bekkara et al.
[13], 30 g of the plant were macerated in 300 mL of
MeOH for 24 h. After filtration and evaporation of the
solvent, the first extraction was obtained with 150 mL
of hot water and 150 mL of ethyl acetate (2 times).The contents of the
mobile phase were filtered before use through a 0.45 um
membrane filter, sonicated and pumped from the solvent
reservoir to the column at a flow rate of 1 mL/min.After the filtration, the
acetone was evaporated and the aqueous layer was
extracted respectively with dichloromethane and ethyl
acetate (2x180 mL).The plant was tested for the
presence of bioactive compounds such as flavonoids,
tannins, anthocyanins, alkaloids, saponins, sterols and
terpenes following standard procedures [9-11].50 g of the dried plant were
macerated in 500 mL of MeOH at room temperature in
dark for 24 h. The solvent was evaporated under reduced
pressure at 60 ?C by rotary evaporator type Buchi R-200
[12].The two organic phases
(ethyl acetate and 1-butanol) were evaporated in a rotary
evaporator device to obtain two phases of flavonoids,
i.e. ethyl acetate and 1-butanol.In this study, the
quantification of some peaks was compared by
calibration of standards: ascorbic acid, gallic acid,
chlorogenic acid, caffeic acid, vanillin, p-coumaric acid
and rutin (Figure 1).The
total polyphenols content was expressed in mg
equivalent of gallic acid (GA) per gram of extract.Sterile paper
discs of 6 mm diameter were impregnated with 10 uL of
various concentrations (0.25, 0.5, 1, and 2 mg/mL) of
each extract [22]. The aerial part of R. raetam was
collected from the Oued Souf region (South-East of
Algeria Sahara).According to the method citing in
Zhang et al. [14], 30 g of the dried plant were macerated
in 300 mL of the water/acetone (7V/3V) in dark and
room temperature for 3 days.The content was expressed in mg
equivalent of quercetin (Qu) per gram of extract.In this work, we used a HighPerformance Liquid Chromatography (HPLC) system,
type Shimadzu LC 20 AL equipped with an universal
injector (Hamilton 25 uL).1 mL of different concentrations of extracts was
mixed with 1 mL methanol containing DPPH (10-4 M)
and incubated in the dark for 15 min.Experimental
2.1.2.2.2.3.2.3.2.2.3.3.2.3.4.2.3.5.2.3.6.