Now heat to 80-85°C over 10 minutes and hold there for at least two minutes but not more than five. Cool in water to room temperature. Transfer the solution to a 100 mL beaker, keep some (~2-3 mL) in the Conical flask. Stir at medium speed on a magnetic stirrer. Now add 30% hydrochloric acid drop by drop till pH of the solution become 7 and filter it.
0.5ml of extract along with 0.1 ml of (0.5N) Folin-Ciocalteu’s reagent was incubated at room temperature for 15 min. After incubation, 2.5 ml of saturated sodium carbonate solution was added and further incubated for 30 min at room temperature. The absorbance was measured at 760 nm. Gallic acid was used as a positive control. Total phenol content was expressed in terms of gallic acid equivalent (mg/g of extracted compounds).The assay was carried out in triplicates and expressed as mean±SD.
For SDS-PAGE, running buffer is prepared by taking 40 ml Tris-Glycine and adding 4 ml of 10% SDS to it. Its volume is raised up to 400 ml by adding distilled water. Load these samples into wells created in the gel. Run the electrophoresis unit at 5 mA and 20 V for entry of sample into lower gel. Once the samples enter the lower gel, alter the current to 10 mA and set the voltage at 90-100 V. Leave this setup undisturbed for around 3 hours.
This was to see the difference between the initial temperature and the final temperature. First Test In a 250ml beaker place 100mls of water, measure the temperature of the water and record this initial temperature onto a table. Set the timer and add one teaspoon of Ammonium Nitrate to the water, stir this continuously until the Ammonium Nitrate has dissolved. After 1 minute measure the temperature and record it, do this for a further 2 minutes (3 minutes in total). Repeat this process for a total of 10 teaspoons.
The measurement was .995g. The salicylic acid was transferred into a 125 ml Erlenmeyer flask and the acetic was given by the lab instructor. Three drops of 85% phosphoric acid. While swirling and mixing more was added and then clamp onto the sing stand in the boiling water for 8 minutes. 1 mL drops of water was added in the flask.
Equal volume of chloroform was added and centrifuged at 13000 rpm for 5 minutes. Again, the aqueous solution was transferred to a new 1.5 ml microcentrifuged tube and 1/10 volume of 3M Sodium Acetate Solution was added. Then, 2.5 volumes of cold absoluted ethanol were added to precipitate the DNA. The mixture was incubated in -20 °C for overnight or -80 °C for 1-2 hour. After incubation, the mixture was then spinning at 13000 rpm for 30 minutes at 4 °C.
2.5.2 Kinetic investigation of FVS in oxidative degradation The kinetics of the acid degradation of FVS were evaluated in 3 % H2O2 at 70°C for different time periods. Solutions containing 1 mg/mL of the FVS were prepared in water. An appropriate aliquot was transferred into a volumetric flask, and diluted with 3 % H2O2 to give final concentration of 100µg/ml FVS. This solution was heated to 70°C, evaluated for time intervals of 30 min, 60 min and120 min. Three samples were analyzed for each time interval.
Also for mucilage extraction, Cactus stems were removed skin and cubed (1 cm3). Samples were homogenised (20% w/v) in distilled water. The slurry was centrifuge for 10 min at 4,500 rpm and the supernatant precipitated in ethanol and finally dried (Sáenz et al., 1992). Preparation of liposomal oil Multilamellar vesicles were prepared according to the thin film hydration method (Gortzi et al., 2006). Lipid solution was prepared by dissolving 5 mg/ml of phosphatidylcholine, 1 mg/ml cholesterol and 0.1 mg/ml essential oil in 3 mg/ml chloroform.
The sample was subjected to steam distillation as illustrated in Figure 1. A total of 50ml of distillate was collected while recording the temperature for every 5.0 ml of distillate. The distillate was transferred into a 250ml Erlenmeyer flask and 3.0 g of NaCl was added. The flask was cooled and the content was transferred into a 250-ml separatory funnel. Then 25.0ml of hexane was added and the mixture was shaken for 5 minutes with occasional venting.
131 The urea and LZM passage was calculated by the following Equation (5): p r C P C (5) 132 where Cp and Cr (mg·L−1) are the concentrations of permeate and remaining solution. The 133 concentration of different solute is determined by UV-Vis spectrophotometer. 134 2.7. Hydrophilic Test 135 The hydrophilicity of membrane surface was characterized on the basis of water contact angle 136 measurement by YH-168A type contact angle goniometer. The membranes (2 × 2 cm) rinsed with 137 ethanol and then put in 60 °C ovens drying for 2 h. The contact angle was measured 10 s after water 138 dropted on the airside surface of membranes.