Preparation of test sample procedure es puter AOAC 969.20, 2007

7 mL H 2 SO 4 were added, also boiling chips which pass No. 10 sieve. Test portion was digested 1 h after all H 2 O is distilled and acid comes to true boil. Minimum volume of H 2 O was added to dissolve solids, cooled, and thin film of petroleum jelly placed on rim of flask. Digested and boiling chips transferred to distillation apparatus and flask rinsed 6 times with 2 mL portions H 2 O. Under condenser with tip extending below surface of solution was placed 125 mL Erlenmeyer contained 5 mL saturated H 3 BO 3 solution and 4 drops indicator Added 10 mL NaOH-Na 2 S 2 O 3 solution to still, collected ca 15 mL distillate, and diluted to ca 50 mL. Titrated to end point. Made blank determination and calculate. Nitrogen content calculated by the equation: Nitrogen wet basis = normality HCl × corrected acid volume mL × . g Nmol weight of wet sample g × Nitrogen dry basis = normality HCl × corrected acid volume mL × . g Nmol weight of dried sample g × Protein = N × Protein factor = N × 5.45 combination of coconut and rice protein factor

3.4 Determination of lipid contents, soxhlet extraction

AOAC 963.15, 2007 Fat flask were dried in a hot air oven at 105 o C for 1 hour and then cooled in desiccator and weighed. Weighed 2 g of sample used fat-free cotton and wrapped by filter paper Whatman® no 1 then tied with a white thread. Placed thimble containing test sample in soxhlet, supported it with spiral or glass beads. Rinsed digestion beaker, drying beaker, and watch glass with three 50 mL portions petroleum ether, and added to wash the thimble. Reflux digested test sample 4 h adjusted heat so that extractor siphons ≥30 times. Removed flask, and evaporated solvent on steam bath. Dried flask at 105°C until it reached its constant weight. Cooled in desiccator to room temperature and weighed. Calculated fat content by the following equation: Fat wet basis = a ‒ b c Fat wet basis = Fat wb ‒ Moisture content wb × Description : a = weight of flask and fat g b = weight of flask g c = weight of wet sample g 2.4.5 Determination of carbohydrate, by difference Carbohydrate content of the sample was calculated by deducting 100 nutrient samples with moisture content, ash content, protein content, fiber content and fat content. Its value could be determined by using the following formula: