Estimation of Fat in Milk Aim: To determine the fat content in milk. Principle: The milk is mixed with H2SO4 and iso-amyl alcohol in a special Gerber tube, permitting dissolution of the protein and release of fat. The tubes are centrifuged and the fat raising into the calibrated part of the tube is measured as a percentage of the fat content of the milk sample. The method is suitable as a routine or screening test. It is an empirical method and reproducible results can be obtained if procedure is followed correctly.
(SDS denatures the proteins, BPB is used as a tracking dye while Beta Mercaptoethanol breaks sulfide bonds.) Heat the samples at 80 degrees Celsius for 5 minutes in a water bath. This helps in protein denaturation. Put the cassette into the buffer dam and fill it with the running buffer. For SDS-PAGE, running buffer is prepared by taking 40 ml Tris-Glycine and adding 4 ml of 10% SDS to it.
Repeat this process for a total of 10 teaspoons. Second Test In a 100ml beaker place 50mls 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.
Bile salts have significant role in lipid metabolism, which emulsify and solubilize dietary fat and fat-soluble vitamins through its detergent activity. Bile salts are deconj... ... middle of paper ... ...onent. The tubes were heated for 10 min at 60°C in a water bath. After cooling, 5 ml of hexane was dispensed to all tubes and vortexed for 5 min at 20-second interval. Then 3 ml of water was added and mixed thoroughly.
It was placed in an ice bath for 10 minutes with occasionally swirls. The third experiment, was Filteration. The vacuum filtration apparatus, the filter flask and the aspirator trap bottle was set up for this section. The crude aspirin was collected by the vacuum filtration. The sample was poured in the Buchner funnel and a about 5 mL of water was in the flask to get the last few of crystal out of the flask.
Measure 90ml sodium thiosulphate into a 100ml measuring cylinder. 3. Using the same measuring cylinder with sodium thiosulphate still in, fill solution up to 100ml with water. 4. Pour solution into conical flask and place conical flask on top of lined paper.
4.) Use pipette to accurately measure 10cm3 of HCl into the 1dm3 volumetric flask. 5.) Add distilled water in a wash bottle to the acid until the level has almost reached the white line, then use a teat pipette to fill it up to the point where the meniscus is just on the line. 6.)
Clean one of the Erlenmeyer flasks using a wash bottle and add 10ml of Acid “A” into it. Use the pipette to accurately measure 10ml before adding it to the flask. 3. Add 2-3 drops of phenolphthalein indicator into to the Erlenmeyer flask containing the Acid “A”. 4.
Protein quantification Five tubes labeled “Blank”, “C”, “LSS”, “HSS”, “HSP” were obtained. The blank contained 100 ul of distilled water. The others had 90 ul of distilled water and 10 ul of the cellular fractions obtained by centrifugation. #ml of BCA were added. The samples were placed in a water bath and incubated at 37 degrees Celsius for 30 min.