3) Centrifuged at 2500 rpm for 12 mins. Upper hexane layer (supernatant) was transferred carefully into another test tube. 4) Evaporated the hexane under a stream of grade 1 nitrogen gas and added 100 µl of methanol to the residue left and vortexed for 1 min. 5) Injected 100 µl of extract in HPLC vials and closed properly. Standard curves and calculations- Retinol was quantified from standard curves peak area for each vitamin.
Two solution of 2.59g Aniline hydrochloride and 5.71g Ammonium per oxydisulphate in 50ml deionized water was prepared separately. Both solution were kept one hour. Then both solutions were mixed. Reaction mixture was stirred for one hour. Then it was provided with stay time of 24 hours.
The end product was passed via sieve (no. 85) and stored in desiccators until use. 2.3. Preparation of DS loaded mucoadhesive beads PC-SA [F0], DS-SA [F1] and DS-PC-SA [F2-F6] beads were prepared by the ionotropic gelation method and the compositions are summarized in Table 1. Initially, PC gum was dissolved in distilled water and boiled for 10 min, cooled and stirred for 24 h at
This reaction is refluxed in a waterbath for two hours and their volume were reduced to 70% of it’s original volume and residue was obtained. The coloured product obtained was filtered under suction, washed with ethanol. The product were recrystallized from ethanol. Their yields ranges from 50-55%, the product obtained were pinkish colour and melting point was 2020C. Step-III: Synthesis of Cr(II) Complexes: The Schiff's base complexes were synthesized by mixing the Schiff's base (1.5 g) in ethanolic solution of Chromium chloride [CrCl2].
The fresh intestine was collected, which was cut through lumen. The microsphers equivalent to 100 mg of the drug was taken on mucosal side of the intestine, was placed b/n donor and receptor compartment; in such a way that mucosal side is facing towards donor compartment. One ml of phosphate buffer pH7.4 was added to donor compartment, the receptor compartment is filled with 25 ml of phosphate buffer pH7.4. The rate of drug permeation was studied by using collection of receptor fluid at regular time intervals, and analysed for drug
Into round bottom flask taken 180 ml nHexane . Extracted at 75°C for 24 h until the solvent leached colourless. Dry solvent from sample. Followed by transesterification Transesterification process Transesterification is the process by which the glycerides present in fats or oils react with an alcohol in the presence of a catalyst to form esters and glycerol Chemical-Catalyzed Transesterification. In a 150 mL Erlenmeyer flask with a magnetic stir bar placed 0.25 g of anhydrous sodium hydroxide and dissolved it with 10 mL of methanol.
Metal chelating activity Briefly, 2 mM FeCl2 was added to different concentrations of test sample and reaction was initiated by the addition of 5 mM ferrozine. The mixture was vigorously shaken and left to stand at room temperature for 10 min. Absorbance was measured at 562 nm after 10 min.8 % Inhibition = [(AB - AA)/AB] x 100, where AB, absorption of blank sample, AA, absorption of test sample. 2.6. Antibacterial
First the solution of 1 mL of extract was added to deionizer water (10 mL), then Folin–Ciocalteu phenol reagents (1.0 mL) added to the mixture. The mixture was left for 5 minutes, and solution of 20% sodium carbonate (2.0 mL) was added to the mixture. This mixture diluted until 50 mL with deionizer water. And after that kept in total darkness room for 1 hour. The mixture absorbance was measured at 750 nm using a spectrophotometer.
This diluted solution will be used in the assay as duplicate samples. Then, 1.0mL of standard glycine solution containing (7.5mg/mL) was diluted to 100mL with water using a volumetric flask. This solution contains 1.0µmole/mL of glycine. 8 tubes were set up according to the following protocol and 2.0mL of ninhydrin reagent was then added to each of the 8 tubes and were placed in a boiling water bath for 20 minutes. After 20 minutes, the tubes from the bath was carefully removed, cooled in a beaker of cold water, then 8.0mL of 50% ethanol was added and mixed well.
The peel powder was soaked overnight (or for desired period) at room temperature (30-32°C) for extraction with intermittent shaking. After extraction, it was centrifuged at 2000 rpm for 5 min and filtered through Whatman No. 2 filter paper. The effective volume obtained after centrifugation was noted. 2.3 Determination of radical scavenging activity (RSA) Determination of RSA was carried out according to Murthy, Jayaprakasha, & Singh (2002) using DPPH as stable free radical and butylatedhydroxyanisole (BHA) as standard.