In 10 g dried sediment sample added 7 ml 0.2 M NH4Cl solution. A mixture of 100 ml hexane: acetone (1:1) was used as a solvent to extract pesticides with overnight shaking for 12 h on reciprocal or wrist action shaker at 180 rpm. The extract was carefully decanted through activated florisil column (2-3 cm), giving twice wash with25 ml hexane: acetone (1:1) to the sediments. The elute was then washed with 200 ml water and then again aqueous layer was extracted with 50 ml hexane. Finally the hexane layer was washed with 100 ml water and then evaporated to dryness with a vacuum rotary evaporator.
Then, we did reflux for 75 minutes. After reflux, we removed the reaction mixture from the apparatus and cooled it for several minutes. We transferred the mixture to the beaker that contained water (30 mL). We cooled the mixture to room temperature and added sodium carbonate to neutralize the mixture. We added sodium carbonate until the pH of the mixture was 8.
Methods Formulation of chloramphenicol ophthalmic hydrogel Formulations was prepared according in Table 1. Poloxamer 188 and poloxamer 407 each weighed and dissolved with distilled water. Then stored in the refrigerator for 24 hours. Next chloramphenicol dissolved with propylenglicol, and nipagin. The mixture was stirred until the entire dissolved and homogeneous.
The inoculum was prepared from fresh overnight broth culture in nutrient dextrose broth. Plates were incubated for 24 hours at 37°C and 96 hours at 28°C for antibacterial and antifungal activity respectively. 3.9 Chromatographic profiling and SDS-PAGE of Peptide/protein extracted from aqueous leaf extract The methods used sequentially for purification of aqueous extract were i) Ammonium Sulfate Precipitation, ii) dialysis and iii) Ion-exchange chromatography on DEAE-Cellulose columns. All steps were carried out at 4ºC to maintain the stability of the isolated products unless mentioned otherwise. The chemicals and dialysis membrane matrix used for the partial purification were ammonium Sulfate, Disodium Hydrogen Phosphate, Sodium Hydrogen Phosphate, Sodium Chloride, dialysis membrane cut off 5 kDa and DEAE-Cellulose.
Then, the test media is then incubated at 37 ° C, for 18-24 hours. Rinsing reusable instruments The samples were rinsed with 40 ml of pyrogen-free water using a glass beaker that is free from pyrogens. Endotoxin testing using STV A total of 0,2mL from the water obtained from the rinsing was placed in the STV containing LAL reagent and was shaken for 20 to 30 seconds. Then STV was placed in an incubator at 37 ° C for 60 ± 2 minutes. STV was then observed by reversing the reaction tube in one smooth motion.
3.3.4 Preparation of CaCl2 solution The molecular weight of CaCl2 was 111 g/mole. Weighed 11.1g CaCl2 and dissolved into 100ml distilled water. The final CaCl2 concentration would be 1M. CaCl2 solution was used to make bottom agar and initiate the infection cycle. 3.4 Selection of most sensitive strain to bacteriophages to make new stock culture Starting culture was prepared by inoculating 1ml (1×109 CFU/ml) stock Lactococcus lactis ssp.
A sample of fish oil (50 mg) was transferred into screw-cap vial. 2 ml benzene and 10 ml sulfuric acid (1%) in absolute methanol were added. The vial was covered under a stream of nitrogen before heating in an oven 90 °C for 90 minutes. Ten ml of distilled water were added to the cooled vial and the methyl esters in each vial were extracted with 5 ml of petroleum ether for three times. The three petroleum ether extracts were combined and concentrate to its minimum volume by using a stream of nitrogen.
A spin vane was added and a water-jacked condenser was attached. Isopentyl nitrite (0.06ml, 0.045 mmol) was dissolved in 1,2-dimethoxyethane (0.50 ml) in a 3-ml conical vial and caped to prevent loss by evaporation. Running the reaction. The mixture in the 5-ml conical vial containing the tetraphenylcyclopentadienone and anthranilic acid was heated on an aluminum block to 140° C. Once the mixture started to boil the prepared mixture of isopentyl nitrite was added to the 5-ml conical vial through the top of the condenser using a pasture pipette. The solution continued to boil for 25 more minutes until a
Commercial TiO2 P25 was obtained from Evonik. Ultrapure water (18MΩ.cm-1) was used throughout the whole experiments. 2.2. Synthesis of photocatalysts The TiO2 nanoparticles were prepared by the sol-gel method described below: 3.9 ml of TiCl4 was slowly added into 10 milliliter of absolute ethanol in reaction vesel, this reaction performed under fume hood at 0°C with vigorous stirring due to exothermic reaction,the high volatilityof TiCl4and also therelease of hydrogen chloride. Then, water was added dropwise during the mixing process.
For chlorination SOCl2 used. We have synthesized carboxamide derivatives using NTAA and 3-(3-benzyl-4,5-dihydroisoxazol-5-yl)pyridine and 3-benzyl-5-(3-nitrophenyl)-4,5-dihydroisoxazole by coupling reaction using dichloromethane as a solvent and the product obtained in 1.5-2 hours by stirring only. After synthesis of these carboxamide derivatives , The compounds are characterized by FTIR, 1HNMR, CMR, UV-Visible and mass spectral analysis. The analysis data is in good correlation with structure of