Each tube was then dragged into the spectrophotometer to be analyzed. A data point for each analyzed tube was placed on the graph to show the optical density and glucose concentrations. After graphing this data, part two needed to be completed. To start, 5 different test tubes were filled with 3 drops of 5 different patients blood followed by 5 drops of deionized water. Next, 5 drops of Barium Hydroxide was placed in each tube to clear proteins and cell membranes for an accurate reading could be made.
4- Set up reflux system using a clean and dry condenser . 5- Place the flask on the hot plate and heat the reaction for 45 minutes - 1 hour . 6- When the reflux is over , remove magnetic stirrer and allow the reaction to cool to room temperature . 7- Add 20 ml of ice water to a separating funnel
pH, Percentage Drug Content and Content drug Uniformity Solution of 1g of gel dissolved in 30mL of distilled water was prepared and pH was determined by using digital pH meter (Systronics 361) and the results were summarized in the table 3. The drug Content Uniformity  was determined by the following method. 1g of gel was dissolved in a 100 mL of phosphate buffer pH 7.4 for 48 hrs with constant stirring using magnetic stirrer. Samples were taken from three different parts of the total ethosomal gel of 1g. Solution was then filtered and observed with UV-spectrophotometer at λmax 254nm.
All the test tubes contained in total 3 mL of solution. The following solutions’ concentrations in a tube were .1 mL of dye and 2.9 mL of water, .25 mL of dye and 2.75 mL of water, .5 mL of dye and 2.5 mL of water, 1.0 mL of dye and 2.0 mL of water, 2.0 mL of dye and 1.0 mL of water, and 3.0 mL of dye and 0 mL of water. These samples were tested by the spectrophotometer, and the absorbencies recorded. This whole process was completed twice and the absorbencies were averaged. Lastly, final concentrations and dilution factors were calculated by using the appropriate formulas.
The brain tissue was harvested and placed in buffered formalin solution. After fixation, the tissue sample was washed thoroughly in running water overnight. It was dehydrated by ascending rates of alcohol (50, 60, 70, 80, 90 and 100%) each for 60 minutes. Then the tissues were cleared with two changes of xylene each with 30 minutes. Embedding was done with two changes of molten paraffin for 30 min to remove xylene.
4) Use a 25 cm3 graduated cylinder for the initial measurements of each solution. Pour 20 cm3 of 0.5M FeCl3 and 20 cm3 of 0.5M KSCN in separate graduated cylinders. 5) Using a 10 cm3 graduated cylinder, pour the necessary volume of distilled water, in this case 5 cm3. After pouring the distilled water to the graduated cylinder containing the FeCl3. 6) Next, prepare a 50 cm3 beaker, and pour both reactants into it.
Then, we did reflux for 75 minutes. After reflux, we removed the reaction mixture from the apparatus and cooled it for several minutes. We transferred the mixture to the beaker that contained water (30 mL). We cooled the mixture to room temperature and added sodium carbonate to neutralize the mixture. We added sodium carbonate until the pH of the mixture was 8.
To clean the syringe, flush it by drawing 6 mL of distilled water. Step 2: Mix both test tubes , shake gently and time the reaction. Step 3: The same step as procedure 1, and step 3 which is to record the observed color step 4: use the palette/color chart to help you identify the observations you make. Safety precautions: Pull your hair back Safety eye goggles Closed toe
The 3 concentrations of enzymes were 0.5 ml, 1.0 ml, and 2.0 ml of turnip extract, while the substrate consisted of 0.1ml, 0.2 ml, and 0.4 ml of hydrogen peroxide. In a separate tube, the control was made up of turnip extract and guaiacol, known as the color reagent. This was recorded the absorbance every 20 seconds for 3 minutes.