The reaction mixture contained 100 µl of each of the extract solution in separate tubes (1 mg/ml) to which was added 0.5 ml of Folin-Ciocalteu phenol reagent, 1.5 ml of 20% (w/v) sodium carbonate and 10 ml of distilled water. After 2h of reaction at ambient temperature, the absorbance was measured at 765 nm and used to calculate the phenolic contents, using gallic acid as a standard. All experiments were performed thrice and the results were averaged and reported in the form of mean ± S.D.Then the total phenolic contents were expressed in term of gallic acid equivalents (mg GAE/g dry extract)
In 10 g dried sediment sample added 7 ml 0.2 M NH4Cl solution. A mixture of 100 ml hexane: acetone (1:1) was used as a solvent to extract pesticides with overnight shaking for 12 h on reciprocal or wrist action shaker at 180 rpm. The extract was carefully decanted through activated florisil column (2-3 cm), giving twice wash with25 ml hexane: acetone (1:1) to the sediments. The elute was then washed with 200 ml water and then again aqueous layer was extracted with 50 ml hexane. Finally the hexane layer was washed with 100 ml water and then evaporated to dryness with a vacuum rotary evaporator.
Then, 3.7 µl of 10 mM Fluorescein Maleimide was added in equal quantities in wild type, mutant and purified ERAB protein, incubated at RT for 10 minutes. Similarly, 4.1 µl of 100 mM DDT was added in the three proteins respectively, incubated for 20 minutes again. And 3 µl of unmodified wt protein was transferred into a 1.5 ml tube as an additional sample. Then, 10.3 µl of 5 x SDS sample buffer was added to each sample and boiled for 5 minutes at 95 oC. Samples were cooled on ice for 1 min, spin down and now they were ready to load on the
The absorbance was measured at 120nm within 10min. Saponin determination The method used was that of Obadoni and Ochuko (2001). The samples were ground and 20g of each were put into a conical flask and 100cm3 of 20% aqueous ethanol were added. The samples were heated over a hot water bath for 4 hours with continuous stirring at about 550C. The mixture was filtered and the residue re-extracted with another 200ml 20% ethanol.
A graduated cylinder was used to add 10ml of distilled water into a 100ml volumetric flask. 5.0ml concentrated acetic acid was pipetted into a 100ml volumetric which already had 10ml of distilled water. After adding acetic acid, the volumetric swirled to thoroughly mix the solution. The volumetric flask was then topped to the mark with distilled water and covered. (2) Titration of Acetic Acid with Sodium Hydroxide 10ml of distilled water was added into a 250ml Erlenmeyer flask.
A flame test and a halide ion test were performed. To start with the flame test, weigh 1 gram of Unknown substance in an analytical balance using a scoopula and mixed it in with 20 mL of water into a 150 mL. Repeat the same step with NaC2H3O2. Repeat the same step for the flame test and record down data. Lastly is the halide ion test.
Firstly, by using pipette, one ml of food homogenate was transferred into a universal bottle containing 9ml of Ringer solution and labelled as 10-2 or dilution 2, 10-1 is dilution 1 which is the food homogenate. 2. Content of the bottle was then mixed using shaker approximately 10 seconds 3. 1ml of 10-2 was pipetted into a new universal bottle containing 9ml of Ringer solution to make 10-3 dilution 4. Again, content of the bottle was mixed by using shaker for approximately 10 seconds 5.
1 ml = 1 µg CN (x) Chloramine –T: Dissolve 1 gm chloramine – T in 100 ml distilled water. Prepare fresh solution daily. (xi) Pyridine (xii) 1-phenyl–3-methyl– 5-pyrazolone solution: Prepare a saturated aqueous solution (approximately 0.5 gm / 100 ml) by adding the pyrazolone to water at 75 0 C. Agitate occasionally as the solution cools to room temperature. (xiii) Bis–Pyrazolone (3,3-dimethyl-1-diphenyl) (4,4’-bis-2-pyrazolone)-(5,5’
The mentioned ratio of the two materials were collected in a quartz crucible. Next, de-ionized water was added into the quartz crucible containing the nickel nitrate and urea. The de-ionized water was poured until it made up 50% of the portion of the solution After that, the entire solution was placed in an oven for about 1 hour. This was done to age the mixture and the temperature was maintained at 80oC. The solution was then taken out of the oven to be put in a pre-heated muffle furnace which was maintained at 650oC.
Fe3O4 nanoparticles were synthesized according to our previously reported method by chemical co-precipitation of Fe2+ and Fe3+ ions with a molar ratio of 1:2 . Briefly, 2.4 g of FeCl3.6H2O and 0.8 g of FeCl2.4H2O were dissolved in 30 mL of deionized water under using continuous N2 purge at 70 °C. and Under vigorous stirring, followed by dropwise addition of 10 mL of NH3.H2O was dropwise added to the reaction mixture until the color of mixture turned to black and kept reacting for 90 min to complete the reaction. At the end, the synthesized Fe3O4 nanoparticles were separated by a magnet and washed by using water and ethanol for further three times with 89.3%