The solution homogeneity expelled, by centrifugation for 10 min. The sample was centrifuged and separated into two layers, and took the top of the sample is injected for HPLC (11,12). Measured concentration in total lipid: The total fats balanced concentration of the pesticide getting by dividing the measured pesticide residue concentration in the overalll tissue sample by the decimal fraction of the sample that consisted of ether-extractable lipid. The total lipid content of each specimen was estimated from its total cholesterol & triglycerides levels by using a summation method. Analytical results for organochlorine pesticides were reported on a lipid-adjusted basis (nanograms per gram or parts per billion) (14).
Then the bank note was washed by either 20 ml Milli-Q water or 5 ml methanol. The bank note was dried and a second extraction was carried out to calculate the percent recovery. At the primary extraction, different concentration of methanol and PBS were examined. 100 %, 90%, 80%, 70%, 50% methanol: Milli-Q water and PBS were the solvent for extraction. The second extraction was mainly by 100 % methanol.
200 µl of lysis buffer (2 % Triton X-100, 1% SDS, 100 mM NaCl, 10 mM Tris-HCl), 1mM EDTA, pH 8.0 and 0.2 g of glass beads were added to each Eppendorf tube. Then 200 µl of the solution phenol/chloroform/isoamyl alcohol (25:24:1) was added to the tubes under the fume hood and tubes were placed on rotator and left to mix for 3 min. 200 µl of TE buffer was added and spun for 5 min at maximum speed, the water phase was transferred to new tubes. 1 ml of cold 96 % ethanol was added, mixed and then spun for 5 min at maximum speed at 4°C. the supernatant was discarded and the pellet re-suspended in 400 µl of TE buffer (40 mM Tris-Base, 20 mM acetic acid, 1 mM EDTA, pH 8.0).
Preparation of ketoconazole loaded Proliposomes Ketoconazole (KTZ) loaded proliposomal gel formulations were formulated by method reported by Perret et al. 1991 with slight modification. Constant amount of drug was added to varying ratios of phophatidylcholine and cholesterol which constitute lipid component of 1mmol quantity. This lipid mixture was prepared in clean and dry, wide mouthed glass vials to which 400µL of absolute alcohol was added and after confirming the formation of homogenous dispersion, the glass vials were heated thermostatically on a water bath at 60-70oC with intermittent shaking. Add 160µL of double distilled water maintained at the same temperature to the transparent solutions formed, these upon cooling change to yellow translucent liquid/gel or white creamy proliposomal gel.
We then added 10cm3 ethanoic anhydride to the salicylic acid and swirled the contents, this mixes together the two chemicals. We then added 5 drops of concentrated sulphuric acid to the flask and thoroughly swirled the mixture, this creates the solution that makes the aspirin. We then warmed the flask for 20 minutes in a 400cm3 beaker of hot water which was approximately 60°C, we made sure the flask did not go above 65°C because this could have caused the contents to evaporate. Part 2: Using a 25cm3 measuring cylinder we measured out 15cm3 of ethanol into a boiling tube and then prepared a beaker half filled with hot water at approx. 75°C, we got this temperature by filling the beaker with cold water and slowly adding boiling water from a kettle until we reached the right temperature.
Using the 0.1 M stock solutions of sugar, a 0.01 M dilution was created for each sugar type by adding water to the stock solution 9 out of 1. A 20 mL dilution was made for each trial. The same volume of each solution (10 mL -5 mL ) was added to the green sponges to create four group; Group 1: Sponge with 10 Ml 0.01 M glucose solution, Group 2: Sponge with 10 mL 0.01 M fructose solution, Group 3: Sponge with 10mL 0.01 M sucrose solution, Group 4 (Control): Sponge with 10 mL water. The four sponges were placed every 90 degrees on the edge of the arena. The isopods were placed into their environment for one to two minutes to acclimate with the environment.
The final volume was adjusted with the same solvent to get concentration of 100 µg/ml. the solution was further diluted to get the concentration of 10 µg/ml, filtered through 0.45 µm filter tips , and aliquots of 20 µl from this solution was injected into HPLC by using an
Response surface methodology is a group of techniques that are used to study the relations between one or measured dependent factors (responses) and several input (independent) factors . The effect of concentration of the three selected variables, ammonium sulphate, yeast extract and 1,2 propylene glycol was studied by this method. The concentration ranges selected for the three factors are listed in (Table 4). All other factors (glucose 10 g/L, glycine 0.2%, temperature 28 °C, initial pH of medium 7.0, agitation rate 200 rpm, inoculum volume 2%) were kept constant. To calculate optimum values of selected three factors, a set of 20 experiments was generated using a 23 full factorial CCD, with six replicates at the centre point, was employed to fit a second order polynomial model.
The aqueous extract was prepared by dissolving 1g of dry extract with 20 ml of sterilized distilled water, so the final concentration of extract would be 0.05 g/ml, from this solution other concentration were prepared (0.1-0.2) g/ml. the solutions were shaken for 30 min. The extract was centrifuged (30,000 rpm; 15 min) and the supernatant was Separated. To hydroalcoholic extract, 80 g of the powder was extracted with aqueous methanol (75%). The other two concentrations were prepared from soaking sixfold aqueous methanol (75%) with different amounts of powder.
Then, putting of choloform in amount of 10 mL and solution of Hanus iodine as amount of 10 mL into conical flask is realized. Addition of these two substance into otheraflask also occurs for blank. Next, waiting for these two samples for exactly 30 minutes is realized. Afterthat, solution of potassium iodine in amount of 15 mL and 40 mL water being distilled are added. Titration of last mixture is performed in company with 0.1 M Na2S2O3 until the obtaining of color in yellow.