The solution was then taken out of the oven to be put in a pre-heated muffle furnace which was maintained at 650oC. This was done for the calcination of the solution. After 2 hours, the quartz crucible was removed from the furnace to be cooled down to ambient temperature. The mixture of nickel nitrate, urea and de-ionized water was grinded for four hours after being removed. Preparation of Nickel Oxide
Pepsin (1 wt. % of the estimated collagen) was added to the supernatant, stirred for another 2 days. Soluble collagen was purified by a repeated process of salting-out with 1 M NaCl, condensation by centrifugation at 14000 ×g for 20 minutes and subsequent solubilization in 0.5 M acetic acid. The collagen solution thus obtained was dialyzed in dialysis tubes (VISKING dialysis tubing, MWCO 12 000 - 14 000 RC, diameter 21 mm) against sterile 0.1 M acetic acid and sterile distilled water, respectively and
The absorbance was measured at 120nm within 10min. Saponin determination The method used was that of Obadoni and Ochuko (2001). The samples were ground and 20g of each were put into a conical flask and 100cm3 of 20% aqueous ethanol were added. The samples were heated over a hot water bath for 4 hours with continuous stirring at about 550C. The mixture was filtered and the residue re-extracted with another 200ml 20% ethanol.
.1.1 CYTOSINE ANALOGUE PREPARATION WITH AROMATIC ALDEHYDE when aromatic aldehyde is used, magnesium is added to anhydrous methanol or ethanol (4 eq relative to cytosine) and heated until complete dissolution of magnesium filings and add 2 mmol of cytosine, followed by the aromatic aldehyde in the amount of 4-6 eq, minimum of 4 eq relative to cytosine, the reaction mixture is heated up to 45-65°C for at least 3 hours, and later, a reducing agent, preferably NaBH(1 eq relative aldehyde) ,is added to the cooled mixture, then it is kept at room temperature for at least 15 minutes, followed by addition of inorganic acid solution; next, the mixture is evaporated, water is added and the mixture is extracted with ethyl acetate to isolate the product;
Incubated at 25℃ for 7 days under the shaking condition. After 7 days, centrifuge it at 5000rpm and at temperature of 4℃ for 20 minutes. Discard the pellet that contains bacterial cells, take supernatant. Make the pH of supernatant of 2 by using "M" "H" _2 "S" "O" _4. Add choloroform and methanol of ratio 2:1 and of equal volume.
Extraction of β-caryophyllene Ground cloves will be used to isolate β-caryophyllene via steam distillation. 5g of ground cloves will be taken in a 500mL RBF(Round bottom flask) with boiling stones, 40mL of dH2O, and 3 to 4 drops of an antifoaming agent (to prevent violent boiling). Then, the contents of the flask will be heated on a heating mantle for an hour and followed by condensation of the distillate through a water jacket. Then allow it to collect in a graduated cylinder. Steam distillation will allow the clove oil to co-distill with the water, which take place at a least temperature than the boiling temperature of the individual solutions.
Synthesis of 2-[(4-methoxy-phenyl)iminomethyl]-4-nitrophenol (SB) The Schiff-base, 2-[(4-methoxy-phenyl)iminomethyl]-4-nitrophenol, (C14H12N2O4) was synthesized by using p-Anisidine in methanol and this was slowly added to 5-Nitrosalicylaldehyde in methanol as shown in Scheme 1. The mixture was refluxed and stirred for 3 hours at room temperature. The completion of the reaction was monitored through TLC for the disappearance of the starting compounds. Then, the solvent was evaporated through rotary evaporator yielding yellow crystals of 2-[(4-methoxy-phenylimino)-methyl]-4-nitrophenol. The yield was about 85%.
(2006), after slight modifications. The fundamental principle of the DPPH method is the reduction of the DPPH radical in an ethanolic solution by an H-donator antioxidant (AH) to form the non-radical form DPPH-H. In a microtube, 10 µL of each fraction at different concentrations (10 - 1000 µg/mL) were mixed with 990 µL of a DPPH solution (0.1mM) prepared daily. The reaction was allowed to develop for 30 minutes in the dark at room temperature, and then the absorbance was read at 515 nm with a spectrophotometer (Spectronic Helios Alpha UV-Visible, Thermo Electron Corporation, U.S.A). The analysis was done in triplicate for each
DMF was used as a solvent and AIBN (0.5% w/w of total monomer) as free radical initiator .The reaction was carried out at 70±2° C for 6 hour with constant stirring. After completion of the process it was cooled to room temperature and resultant polymer solution was poured in the large amount of methanol with stirring when polymer precipitated out. It was filtered and washed with methanol. The polymer was purified by repeated precipitation using methanol from solution in DMF and then it dried. 2.3 Preparation of PS
(Voucher specimen No. 1246). Propionibacterium acnes Strain (MTCC 1951) was procured from (MTCC) IMTECH, Chandigarh. Preparation of extracts and fractions The plant materials were dried and ground to a coarse powder. The ground plant powder (300 gm) was extracted with (80%) methanol by soxhelation for 48 hours.