Add choloroform and methanol of ratio 2:1 and of equal volume. Mix the mixture and left it for overnight for the process of evaporation. The white coloured sediment appeared that is the biosurfactant. Drying of biosurfactant Poured the sediment in the sterile petriplate of known weight. Put the plate in the Hot Air Oven at 100℃ for 30 minutes.
Add 50 to 100 ml of freshly neutralized hot ethyl alcohol and about one ml of phenolphthalein indicator solution. 4. Boil the mixture for about five minutes and titrate it against the standard alkali solution while shaking vigorously during the titration. The weight of the oil taken for the estimation and the strength of the alkali used for the titration shall be such that the volume of alkali required for the titration shall be such that the volume of alkali required for the titration does not exceed 10 ml. Calculation: Acid value= 56.1VN W Where, V=Volume in ml of standard potassium hydroxide or sodium hydroxide used N= Normality of the potassium hydroxide solution or sodium hydroxide solution; and W=Weight in g of the
They were chopped into small pieces and homogenized using a blender for 2 min. The homogenized samples were kept in the freezer maintained at -8000C for three days. Later, all the samples were grounded into fine powder using a dry grinder and stored in a freezer at -2000C before extraction. Ten grams of samples were homogenized in 250 ml 80% (v/v) methanol at room temperature. The mixture was shaken using shaking incubator at 200 rpm for 120 min at 5000C.
Pepsin (1 wt. % of the estimated collagen) was added to the supernatant, stirred for another 2 days. Soluble collagen was purified by a repeated process of salting-out with 1 M NaCl, condensation by centrifugation at 14000 ×g for 20 minutes and subsequent solubilization in 0.5 M acetic acid. The collagen solution thus obtained was dialyzed in dialysis tubes (VISKING dialysis tubing, MWCO 12 000 - 14 000 RC, diameter 21 mm) against sterile 0.1 M acetic acid and sterile distilled water, respectively and
DMF was used as a solvent and AIBN (0.5% w/w of total monomer) as free radical initiator .The reaction was carried out at 70±2° C for 6 hour with constant stirring. After completion of the process it was cooled to room temperature and resultant polymer solution was poured in the large amount of methanol with stirring when polymer precipitated out. It was filtered and washed with methanol. The polymer was purified by repeated precipitation using methanol from solution in DMF and then it dried. 2.3 Preparation of PS
Measurement of lipid peroxidation TBARS, a measure of lipid per oxidation, was measured as described by Ohkawa . Briefly, 1 ml of suspension medium was taken from the 10% tissue homogenate. 0.5 ml of 30% Trichloroacetic acid (TCA) was added to it, followed by 0.5 ml of 0.8% thiobarbituric acid (TBA) reagent. The tubes were covered with aluminium foil and kept in shaking water bath for 30 minutes at 80°C. After 30 minutes, tubes were taken out and kept in ice-cold water for 30 minutes.
Dialysis membrane having a pore size 2.4 nm and molecular weight cut off between 12,000 and 14,000 was used. The membrane was soaked in double distilled water for 12 hr. before mounting in a Franz diffusion cell. About 1ml of semisolid preparation of NLC was applied to the donor compartment, and the receptor compartment was filled with phosphate buffer, pH 7.4 (14 ml). During the experiment, the solution in the receptor side was maintained at 370C and stirred at 800 rpm with Teflon coated magnetic stirrer bar.
Diphenylcarbazide was added to the solution and allowed to stand 10 minutes for full color development. The absorbance was measured at 540nm using spectrophotometer (Gilcreas et al., 1965). Batch biosorption studies and adsorption isotherms Batch biosorption studies were carried out in 100 ml of Erlenmeyer flask containing 25 ml of potassium dichromate solution as source of Cr (VI) and agitated in an orbital shaker at 160 rpm. The effect of different parameters such as initial metal concentration, adsorbent dosage, temperature, contact time and pH were investigated. All experiments were carried out at pH2 and temperature 37 ◦C.
Then, the solution was loaded into the well. The gel then was run at 100 volt for 30 minutes. After that, the agar was cleaned and put into the EtBr solution for 10 minutes. The agar then washed by using tap water. Next, the agar was observed under UV transilluminator to see the formation of band.
It has been prepared with some modifications. ZP was prepared as follows: typically, 5 g ZrOCl2.8H2O was refluxed with 50 ml of 12 M H3PO4 at 100 °C for 24 h. The obtained precipitate was filtered off and washed with 0.1 M H3PO4 until free of chloride ion. Finally, the solid was washed with distilled water several times until the neutral pH and dried in an oven at 110 °C for 24 h. The final product was ground into fine powders and confirmed by XRD (Fig.1). The ZPFe was prepared through an ion-exchange reaction[50, 51] (. 3 g of ZP was dispersed into the 50 ml deionized water at 50 °C.