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.
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.
By calculating the Rf values for each compound and comparing them to the known lipids, we are able to distinguish the lipids within the grounded nutmeg. II. Procedures Add 5 g crushed nutmeg and 50 mL hexane-isopropanol into a flask and warm for 15 minutes. Remove the extra solvent on a steam bath under a hood while flushing the flask with N2 gas, leaving the crude extract. Weigh extract.
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.
One g of material was ground and extracted with 0.2 N HCl by continuous shaking. To 0.2 ml of the filtrate, distilled water to make volume 1.4 ml was added. After that1ml of ferric ammonium sulphate solution (21.6 mg in 100 ml water) was added, mixed and placed in a boiling water bath for 20 min. The contents were cooled and 5 ml of isoamyl alcohol was added and mixed. To this, 0.1 ml ammonia solution was added, shaken thoroughly and centrifuged at 3000 rpm for 10 min.
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.
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. Determination of flavonoids The amount of total flavonoids in the acetone extract of seeds of M.zapota was determined by aluminium chloride colorimetric method . The reaction mixture 3 ml consisting of 1 ml of sample (1 mg/ml), 0.5 ml of (1.2%) aluminium chloride and 0.5 ml (120mM) potassium acetate was incubated at room temperature for 30 min. The absorbance of the reaction mixture was measured at 415 nm. Quercetin was used as positive control.
Tube #3: 8mL 5% starch, 1mL pH 7 solution, 6mL salt concentration, 0mL water, and 1mL of amount of enzyme starting. Tube #4: 8mL 5% starch, 1mL pH 7 solution, 0mL salt concentration, 6mL water, and 0mL of amount of enzyme starting. Then at time zero remove 1 mL of the solution from each of the tubes as the baseline control. Place this solution in a tube placed at time zero and continue to add 2 mL of water and 3 drops of potassium iodide. Add 1 mL of amylase to the stock solution of substrate/ salt solution.
Characterization of Aspirin. In the first section, the Synthesis of Aspirin, salicylic acid was weight to be 3.029 grams using mass by difference since it was weighed on a 150 milliliter beaker. 9.23 milliliters of the acetic anhydride and 14 drops of 85 percent phosphoric acid were added to this beaker. A Bunsen burner provided by the laboratory was then used to boil the just mixed combination by producing a flame underneath the positioned beaker on top, and then allowed to cool for several minutes after the Bunsen burner flame was terminated. Two quantities of distilled water were then added to this mixture to make it cool even further, which were 41 drops and 30 milliliters.
100 uL of this solution was set aside in a tube labeled “HSS.” SDS-PAGE preparation. Eight microfuge tubes were taken and divided into two sets of four, labeled A and B. Each set had was labeled like the four we prepared earlier, “C”, “LSS”, “HSS”, “HSP.” 25 ul of the respective solution was pipetted into the two sets of tubes. Set A was given 25ul of 2X Laemli buffer A. Set B was given 25 ul of Laemli buffer B. Laemli buffer ( .125M Tris HCl, pH 6.8, 20% glycerol, 4% SDS, and 10% BME).