1.00 ml 10 g Ni (iii) Hydrochloric acid (L.R. grade) 1.0 N : Dilute 100 ml concentrated hydrochloric acid to 1000 ml with grade I distilled water. (iv) Bromine water : Saturate distilled water with bromine to prepare Bromine water. (v) Heptoximereagent : Dissolve 0.1 g 1,2 –cycloheptanedionedioxime (heptoxime) in 100 ml 95 % ethyl alcohol. (vi) Sodium tartrate solution : Dissolve 10g sodium tartrate (Na2C4H4O6.2 H2O) in 90 ml distilled water.
Apparatus- Chromatography column: C18 (10 microns particle size), with Guard column Flow rate: 1.2ml/min Pressure: 30-40kgf Wavelength: 326nm Mobile phase: methanol : water (95:5 v/v) Internal standard: retinyl acetate Injection volume: 20µl Procedure for Retinol extraction from serum samples- 1) 100 µl of serum sample and 100 µl of Retinyl acetate were added into 12 X 100mm glass test tubes. Vortex-mixed for 30 seconds. Then, kept them at 4 C for 5 mins. 2) 1mL of hexane was added and vortex-mixed intermittently for 60 sec. 3) Centrifuged at 2500 rpm for 12 mins.
A 50 mL buret was obtained and was washed with NaOH solution. After filling the buret with NaOH (titrant) and preparing the KHP (analyte) in the Erlenmeyer flask, the solutions were titrated. The volume used from the NaOH solution was recorded. C. Determination of the Acidity of Soft Drinks First, the soft drinks were heated. Upon cooling, it was shaken until no bubbles were formed.
Then, the flask is put on shaker table and mixed at 150 rounds per minute before allowing them to settle for 10 minutes. After settling, the water sample is poured from side spout which is connected to the bottom of the flask. Some researchers reported better reproducibility with modified flask where stopcock is installed at the bottom of the flask, instead of side spout to pour the water sample (Blondina, et al., 1997) (Sorial, et al., 2004a) (Sorial, et al., 2004b). The dispersed oil in the removed water sample is extracted into methylene chloride for further analysis. Then, the oil concentration is evaluated using 340, 370 and 400 nm light absorbance (Environmental Protection Agency,
Absorbance of the solution was measured at 238 nm. Drug content was calculated by the formula absorbance/ slope* dilution factor. Drug loading Microsphers equivalent to 100 mg of DLTZH/ND taken and washed with 3 x 10 ml of methanol, which removes the free unloaded drug. Filtrate was diluted suitably to Beer’s concentration range and free drug concentration was determined spectrophotometrically at 238 nm and 236 nm for DLTZH and ND respectively. Further microspheres were crushed, dissolved in methanol and made up to 100 ml, which was further diluted suitably to Beer’s rangeand drug concentration was determined.
Subsequently extracted by Microwave at power level 70 for 16 minutes and then filtered. The filtrate obtained, was added 10% HCl (until pH 2-3). Then do the bleaching with NaOCl diluted with water 1: 1 to white. Then converted to sodium alginate by adding 20 g of Na2CO3 and stirred in a mixer. The resulting solution is then etched with ethanol to form sodium alginate fibers.
When a carboxylic acid and alcohol was being mixed an ester was formed. In this first reaction (part A) the acid used was salicyclic acid while the alcohol that was used was methyl alcohol. A small amount of concentrated sulphuric acid about 3-4 drops was added and used as a catylst. Sulphuric acid helps to remove the water from the products formed. The mixture was placed in a water bath (90ºC) for 10 minutes to allow the reaction to be completed.
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
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.
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]. 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 light green colour and melting point was 2100C.