1 “substrate” and another “ enzyme.” Instead of using the distilled water, this time you are going to use different pH buffer in the enzyme test tube. In the substrate tube, add 7 mL of distilled water, 0.3 mL of hydrogen peroxide, and 0.2 mL of guaiacol for a total volume of 7.5 mL. For the enzyme tube, instead of distilled water add the pH solution (3) and 1.5 mL of peroxidase which equals a total volume of 7.5 mL. Use the dH2O syringe for our pH solution. To clean the syringe, flush it by drawing 6 mL of distilled water.
The supernatant was assayed for SOD activity by following the inhibition of epinephrine auto-oxidation. 0.5ml of sample was diluted with 0.5 ml of distilled water, to this 0.25 ml ethanol, 0.5 ml of chloroform (all reagents chilled) was added. The mixture was shaken for 1 min and centrifuged at 2000 rpm for 20 min. The enzymatic activity in supernatant was determined. To 0.05 ml of carbonate buffer (0.05 M, pH 10.2) and 0.5 ml of EDTA (0.49 M) was added.
50 mL of distilled water was approximately added to this 250 mL beaker and gently swirled so that the solid (potassium acid phthalate) got fully dissolved into the water. 2 drops of phenolphthalein indicator solution was added to the beaker. The pH electrode was calibrated using the pH buffers. For this, 25 mL of pH 4 and pH 7 buffers were taken in 50 mL or 100 mL beakers. The buffer solutions were saved in case the electrode needed to be re-calibrated later on.
The chemical equation for this experiment is hydrochloric acid + sodium thiosulphate + deionised water (ranging from 25ml to 0ml in 5ml intervals) sodium chloride + deionised water (ranging from 25ml to 0ml in 5ml intervals) + sulphur dioxide + sulphur. As a scientific equation, this would be written out as, NA2S2O3 + 2HCL + H2O (ranging from 25ml to 0ml in
Biuret test is adopted to quantify proteins in fluid by using a spectrophotometer. The biuret solution is a blue solution made up of sodium hydroxide and copper (II) sulfate which turns pink or violet in the presence of proteins, peptides and compounds containing 2 or more peptide linkage. A spectrophotometer measures the respective amounts of light consisting of different wavelengths absorbed and transmitted by a pigment solution. Spectrophotometer works when white light is separated into lights of different wavelengths by a prism which different colors of light passes through the sample. The transmitted light strikes a photoelectric tube, converting light energy to electric current that is measured by a galvanometer.
The sodium thiosulfate was standardized by titration with potassium dichromate dissolved into 50ml that has 1ml H2SO4 and 0.2g Na2CO3 and 0.5g potassium iodide that was allow to stand a while. The solution was titrated until the yellowish color was completely disappeared. Then 1ml of starch solution is added it became dark blue. The titration with thiosulfate was then continue until the dark blue color was disappear making the solution colorless. In the preparation of 2.4M manganese(II)sulfate solution 20.5g of MnSO4 ▪ H2O was dissolved in 50ml distilled water then it was transferred to amber bottle.
Inoculated the Lactococcus lactis ssp. lactis C2 culture onto the petri dish with bottom agar by using inoculation loop for streaking. The streaking was used to isolate the pure strain of Lactococcus lactis ssp. lactis C2 from the stock culture. The inoculation loop was sterilized in a fire for 5-7 seconds and then cooled to dip into the starting culture for microbes inoculation on the plate.
In the donor compartment, proliposomal formulation and control (drug suspended in sodium CMC) equivalent to 10mg of KTZ were placed separately. 30% v/v acetonitrile containing phosphate buffer saline (pH 7.4) was taken as receptor medium and kept under constant stirring upto 24 h. In order to avoid evaporation of the contents, donor compartment and sampling port are covered with aluminium foil. 500 µL of aliquots are drawn at predetermined time intervals and equal volumes have been replaced so as to maintain receptor phase volume. All the samples withdrawn were estimated for drug content using HPLC and the data was fitted into mathematical equations (zero order, first order and Higuchi models) (Szuts et al., 2010) to derive the kinetics and mechanism of drug release from proliposomal gel
The DEAE CL-6b gel was washed twice with 0.5m Hcl, twice with 0.5m Naoh and twice with PB ph6.0 before utilization. For DEAE 1ml gel every 1ml serum was utilized. In the wake of blending for 1 hour at 200c the suspension was centrifuged at 4500g for 25 minutes. The supernatant was thought by Ammonium sulfate precipitation and pellet was broken up in refined water and the ensuing arrangement was dialyzed against PBS ph7.2 to evacuate Ammonium sulfate and conformed to the first volume with yhe same cushion (Hong et.al., 1994). 2)
The spectra were recorded by making dried KBr pellet. A total scans of 16 scans per sample at resolutions were obtained over mid IR region of 400 to 4000 cm-1. The percentages of the diterpene glycosides were determined by High Pressure Liquid Chromatography (HPLC). For mobile phase, HPLC grade acetonitrile and water (80:20) were mixed (pH of 3.0) and filtered through 0.22 µm Millipore filter. Accurately weighed 50 mg (dried at 1000C, 3h) of stevioside standard was taken in a 100 ml volumetric flask and diluted to volume with mobile phase.