Whatman filter paper number 3

CFE was prepared as described previously [73 (link),74 (link)], Briefly, 500 gm of clove flower buds were cleaned from the impurities and washed by distilled water, then dried in a hot air oven (Alexandria Co., Alexandria, Egypt) for 24 h at 40 °C. The flowers were then grounded to powdered form using a clean sterile mortar and pestle (Moulinex, Cairo, Egypt) and packed in an airtight plastic container until used. About 10 g aliquots of the powdered clove flower buds were extracted with 1000 mL ethanol by maceration, then filtered through a Buchner funnel with Whatman filter paper number 3, then evaporated under reduced pressure to dryness at 45 °C. A stock preparation (1 g/50 mL) of CFE was suspended in 1% DMSO and sterilized by Milipore filtration (0.45 μm, Amicon). The plant phytochemical screening was done with GC-Mass [29 (link)]. A direct capillary column TG-5MS (30 m × 0.25 mm × 0.25 μm thickness) was used. About 3 μL of CFE was injected automatically to the equipment using an Auto sampler AS3000 coupled with GC in the split mode. Then the instrumental analysis was carried out as described previously [11 (link)]. The components were identified by comparison of their retention times and mass spectra with those of Wiley 09 and NIST 11 mass spectra databases [11 (link)].

Complete dry fruit R. capitata was pulverized with a mortar. Then, 1 g of the powder was placed into a flask (100 mL capacity), dissolved in 20 mL of a 1:1 water/ethanol mixture, and sonicated for 30 min at room temperature. Separation of solid and liquid phases was performed in a vacuum line system with a funnel. Whatman filter paper (number 3) and fiberglass were used to retain most of solid particles and to allow a faster filtration. The liquid phase obtained was reserved inside a ball flask, and subsequently the solution was concentrated under reduced pressure to isolate the R. capitata extract (a semi-solid), which was stored at 4 °C.