We added sodium carbonate until the pH of the mixture was 8. After neutralize, we collected benzocaine by vacuum filtration. We used a Buchner funnel to collect benzocaine. We used three 10 ml of water to wash the product. After the product was dry, we weighed, calculate the percent yield and determined the melting point of the product.
Then the blower was turned on for sufficient duration and UV lights were switched on for one hour. LAF microbial contamination test (Settling plate method) A total of three petri dishes were prepared aseptically inside a laminar air flow (LAF), and then the petri dish was filled by pouring sterile Tryptone Soy Agar (TSA) liquid media and allowed to solidify. One test media was placed in the LAF cabinet, one test media is placed beside the LAF cabinet while the other the test media was placed near the door. The three petri dishes lid were opened and allowed to stand for 15 minutes and closed again. Then, the test media is then incubated at 37 ° C, for 18-24 hours.
10- Transfer the ester layer to a small dry test tube and dry the ester with anhydrous CaCl2 and stir for 10 min. 11- put it in a preweighed dry round bottom flask . 14- Determine the yield, refractive index, and % yield of ester. Conditions :- 1) This reaction is catalyzed by acid, Like Fischer esterification. 2) Usage of water in step (5):So that after Estrification is completed , any excess unreacted acetic anhydride is hydrolyzed.
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.
The mixture was stirred until the entire dissolved and homogeneous. The materials were ready each put in a bottle 100 mL size vial, then sterilized with autoclave for 15 minutes at 121 °C. The preparations hydrogel done within aseptic at LAF cabinet. Evaluation of chloramphenicol hydrogel ophthalmic preparations Organoleptic test Organoleptic hydrogel checked by observing changes in color, odor and clarity. Clarity was checked visually by examination
Put 1 capsule of the bioflorin into each of the test tubes and tap them until the capsule dissolves. Screw the liquid onto both test tubes to make sure that they are sealed. You now have to wait for approximately two days, in order to obtain satisfying results. Light the candle/put it on fire. Fill the third test tube with approximately two millimeters of Ethanol.
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.
This organism was cultured under solid-state fermentation for 72 h using wheat bran as the substrate. After 72 h, crude enzyme was extracted from the culture medium. The fibrinolytic enzyme was purified from the crude sample by various steps: ammonium sulfate precipitation, dialysis, ion exchange chromatography, and casein-agarose affinity chromatography. All purification steps were performed at 4°C until otherwise stated. The crude enzyme was precipitated at 70% saturation of ammonium sulfate, and the protein was collected by centrifugation (10,000×g for 10 min).
Dissolve the salt in 60 ml of tap water. Add 30 ml 6 M Hcl and stir the mixture with a glass rod. Add 12 g solid Nacl to the solution and stir the mixture for about 2 minutes. Support a 250 ml separatory funnel on a ring, making sure that the stopcock is closed and that a clean beaker is placed beneath the exit tube. Transfer the aqueous solution from the beaker to the separatory funnel.
One millilitreer of the lichen extract (1 mg/mL) in a volumetric flask was diluted with distilled water (46 mL), and the content was mixed in a volumetric flask after adding. oOne millilitreer of Folin-Ciocalteu reagent was added and the content of the flask was mixed. thoroughly . After 3 min, 3 mL of 2% sodium carbonate (2 %) was added and then was allowed to standleft for 2 h with intermittent shaking. The reaction absorbance of prepared mixture absorbance was measured at 760 nm in a spectrophotometer (Jenway UK).
Once the gel hardened, .5X TBE (44.5 mM Tris base, 44.5 mM boric acid, and 1.0 mM EDTA) was added just until the gel was covered with the TBE buffer. Each sample was loaded into the gel as well as 10 μL of DNA size markers (1kb ladder, New England Biolabs) into a separate lane. The gel was allowed to harden at room temperature and then electrophoresed at 100 volts for 75 minutes. Using a UV imager, a photo was taken of the resulting traveled DNA fragments in the gel. Results Table 1.
The dried roots of Inula racemosa were pulverized and sieved with 100 ~ 200 mesh. The herb powder was placed into a glass bottle. Ultrasound-assisted extraction was carried out in an ultrasonic cleaner RK102H (Bandelin sonorex, Germany). The powder of Inula racemosa was extracted three times under the following conditions: the ratio of material to solvent was 10:1, undergoing ultrasonic treatment 30 minutes at 25 °C, 100 kHz /450 W.31 Before large extraction, a small-scale extraction experiments were carried out: 95% ethanol and ethyl acetate as the extractive solutions was investigated, respectively. The extraction efficiency was evaluated according to the percent content of AL and IS contained in the dried roots of Inula racemosa and calculated