3) Centrifuged at 2500 rpm for 12 mins. Upper hexane layer (supernatant) was transferred carefully into another test tube. 4) Evaporated the hexane under a stream of grade 1 nitrogen gas and added 100 µl of methanol to the residue left and vortexed for 1 min. 5) Injected 100 µl of extract in HPLC vials and closed properly. Standard curves and calculations- Retinol was quantified from standard curves peak area for each vitamin.
The crude enzyme was precipitated at 70% saturation of ammonium sulfate, and the protein was collected by centrifugation (10,000×g for 10 min). It was suspended in minimal volume of double-distilled water. The precipitate was dialyzed and used for enzyme purification using column chromatography. The dialyzed sample was loaded on DEAE-cellulose column, which was equilibrated previously with the buffer A (50 mM sodium phosphate buffer, pH 7.4). The column was washed with the five bed volumes of buffer A, and the enzyme was eluted with buffer A
Absorption was read by spectrophotometer at wave length (510CHOL) and (505TG). 6. After this we applied the following equation Result = ×CON. standard Extraction of tissue The extract one gram of fat tissue using n hexane, resolve to dissolve tissue homogenizer homogeneity by adding 1 ml of the same solution to become a 3: 1 and continued homogeneity of the sample to become a solution homogeneity. The solution homogeneity expelled, by centrifugation for 10 min.
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
Approximately 1µg Mbgl was used with 5 mM 4-nitropheny-β-D-glucopyranoside (PNPG) in the reaction mixture of either 2 ml or 1 ml. The reaction was stopped by adding an equal volume of 0.2M Na2CO3and the released product 4-nitrophenol was quantified based on the millimolar extinction coefficient of 18.1 mM-1cm-1at 400 nm (Workman and Day 1982). The optimum pH was determined using the same assay in the 100 mM phosphate-citrate buffer in the pH range of 3.0 to 7.0 and for pH 8.0; 100 mMtris buffer was used. Similarly, the temperature optimum was determined in 100mM citrate buffer of pH-6.0. Thermal stability was determined by incubating the protein solution at 50°C and 55°Cfor different times followed by standard PNPG assay, as described earlier.
Know how to use an electronic balance, calibrate glassware, and know how to use a volumetric flask, calculate density and know how to calculate the percent error. The first thing we need to do is determine the best procedure for measuring density. The materials needed are going to be a 10mL volumetric flask. Conduct 3 separate trials where we will measure the density between different salt solutions. In this experiment we have 6% salt solution, 12% salt solution, 18% salt solution and 24% salt solution.
In the round-bottom flask (100 mL), we placed p-aminobenzoic acid (1.2 g) and ethanol (12 mL). We swirled the mixture until the solid dissolved completely. We used Pasteur pipet to add concentrated sulfuric acid (1.0 mL) to the flask. We added boiling stone and assembled the reflux. Then, we did reflux for 75 minutes.
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.
The analysis was carried on C18 shim- pack GIST (150mmx 4.6mm 5µ) column used as stationary phase. A freshly prepared mobile phase consisting of methanol: potassium dihydrogen phosphate buffer in ratio of (30:70 v/v), PH-3 adjusted using ortho phosphoric acid (OPA) these were filtered by 0.45µM Whatmann filter paper and sonicated before use. The flow rate of mobile phase was 1ml/min. The detection was carried out at 220 nm and run time was around 10 minutes. Selection of wavelength A UV spectrum of drotaverine hydrochloride, ethamsylate, tranexamic acid in water was noted by scanning the solution in the range of 200-400nm.
0,1 gr of kaffir lime oil nanocapsule is weighed and diluted to 100 ml using aquadest, taken as much as 1 ml (100x dilution) to put in the reaction tube then 1ml of saturated NaCO3 solution is added to the test tube and incubated for 10 min at room temperature. Then 0.5 ml of the folinciocalteu (Chemix CV, Yogyakarta) reagent and 7.5 ml aquadest were added, the mixture homogenized using vortex and then incubated for 30 min at room temperature under dark environmental conditions. Absorbance of the sample was then measured using a UV-vis spectrophotometer at a wavelength of 770 nm. The total phenol content of the sample was interpreted to be equivalent to gallic acid based on the standard curve of obtained gallic