Approximately 2 gm, nearest to 0.1 mg, oven dried cornhusk fibres, were weighed out accurately in weighing bottle and transferred to a 100 ml beaker. 40 ml of cold (10-15˚C) 72% sulphuric acid was added gradually to the fibres in small increments while stirring the mixture and macerating the fibres with a small glass rod. The beaker was kept in a bath at 2 ± 1˚C for dispersion of material. After the specimen was dispersed, beaker was covered with a watch glass and kept in a bath at 20 ± 1˚C for 2 hours. Mixture was stirred frequently to ensure complete
Next, a basic stock solution was used to prepare various concentrations ranging from 1.0 x 10-8M to 1.0 x 10-1M by serial dilution. The tissue was washed by overflow with reservoir’s solution for 5 seconds to obtain baseline before adding 0.1ml, 0.3ml and 0.5ml for each concentration respectively into the tissue bath.The tissue’s peak response for each final bath concentration(FBC) was measured and recorded. Rmax and EC50 of histamine were recorded. Later, 5ml of 1 x 10-6 M of mepyramine was added into the reservoir containing 1000ml of Krebs-Henssleit solution to produce a FBC of 5.0 x 10-9M. It was equilibrated with tissue for 10 minutes by flushing into the organ bath.
Once a prevalent color change had been observed at approximately 4 minutes (blue green color) the crucible was set on the counter using the tongs to cool for approximately 5 minutes. The appearance after this period resulted in another color change back to white. The crucible, lid, and hydrated copper sulfate was weighed again to calculate the mass of water lost by dehydration (described in table 1.3). This was done by subtracting the final mass by the initial mass of the crucible, lid, and compound. The mass of the crucible would remain unchanged while the mass of the compound would be altered.
Then five millilitres of sample “A” were placed in the test tube labeled “A”. This was then repeated with the next three samples. Each sample was visually observed and the colour of each was recorded. Next 20 drops of Benedict’s solution were added to each test tube and the test tubes were lowered into a hot bath at a temperature of approximately 80 degrees Celsius. All colour changes were recorded.
Heat the Agarose gel in a 65 °C water-bath to melt the agarose. After it melted, maintain its temperature at 55 °C until it is ready for use. 2. Transfer the spheroids from the 96-well plate to a 15 mL centrifugal tube using a 1000 μL pipette 3. centrifuge the tube for 5 min at 1000 rpm to form a pellet. 4.
The chart shows the length, width and area of each of the eight carbon fiber pieces. To clean the carbon fiber pieces, soak the carbon fiber pieces into 1 M sulfuric acid for 10 minutes. This process can increase its hydrophilic property (oxidizing) and clean other chemicals on its surface. Then dip the carbon fiber pieces into ethanol for 5 minutes in order to clean them
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.
During this step, I observed that there were bubbles in the solution, especially at the bottom of the beaker. After adding the HLC, there solution had a slight yellow tint. Next, I mixed 0.529g of sodium acetate in 3mL of water and added 0.679g of acetic anhydride to the aniline solution and immediately added sodium acetate. The solution was cooled in an ice bath for fifteen minutes. During this time, I noticed the formation
After the specified time, the solutions were completed to mark by using distilled water to achieve a final concentration of 20 µg/mL each, filtered and injected into the HPLC system. Dry Heat Degradation For thermal stress, 10 mg portions of each of VAL and SAC dry powder were placed in porcelain dish in a controlled-temperature oven at 100˚C for 4 hours. After the specified time, the content of the porcelain dish was transferred quantitatively with HPLC-grade methanol into a 10 mL volumetric flask and the volume was made to the mark by using the same solvent. Then, an aliquot of this methanolic stock was diluted to volume with distilled water to obtain a final concentration of 20 µg mL-1. After the preceding treatments, solutions were filtered and injected into the HPLC
Preparation of hot water extracts: 20 g from each bark powder was surface sterilized and mixed with 100 ml sterilized water and placed in water path for 90 minutes at 100ºC, and then filtered through 3 layers of sterilized cheese cloth. The products were kept in a refrigerator at 4 ºC until needed for testing their bacterial
After adding 20 microliters of proteinase K to the 1.5 ml tube, the sample was vortexed for 30 seconds to disrupt the pellet. The sample was then incubated at 56°C to lyse the tissue. The sample was checked every fifteen minutes and vortexed between each checking for an hour and a half until the tissue was completely lysed. The tissue sample was then again vortexed. Next 200 microliters of buffer AL was added and
If the color changes, then it is known that starches are present in the solution. Proteins are next tested. In order to do this, 5 mL of gelatin solution is added to the test tube. 10 drops of Biuret’s reagent are added to test for protein. If there is a color change, then it is known that protein is present in the solution.
The silver ion TLC was prepared through the following procedure: Silver nitrate was dissolved in 10 ml of distilled water. This aqueous solution of silver nitrate was absolutely mixed with 9 g of silica gel (10 ~ 40 μm particles). Then, a 10 × 5 cm TLC plate was coated with the above slurry and activated for 1 h at 90 °C before use. They were immediately transferred into a desiccator in dark for storage after cooling. 32 100 μL of afore-prepared sample solution and the mixed reference standard were diluted 100 times with ethyl acetate.
Once the gel hardened, .5X TBE (44.5 mM Tris base, 44.5 mM boric acid, and 1.0 mM EDTA) was added just until the gel was covered with the TBE buffer. Each sample was loaded into the gel as well as 10 μL of DNA size markers (1kb ladder, New England Biolabs) into a separate lane. The gel was allowed to harden at room temperature and then electrophoresed at 100 volts for 75 minutes. Using a UV imager, a photo was taken of the resulting traveled DNA fragments in the gel. Results Table 1.