Crude Fat AACC International Method Crude Fat in Flour, Bread, and Baked Cereal Products Not Containing Fruit Final approval April 13, 1961; Reapproval November 3, 1999 Objective Crude fat is determined by acid hydrolysis of the sample with HC1 followed by extraction of hydrolyzed lipid materials with mixed ethers. Bthers are evaporated, and lipid residue is heated to constant weight at 100°. Residue is expressed as % crude fat. This method is applicable to cereal products, particu- larly heat-treated products, but not to cereal products containing fruit. Apparatus 1, Erlenmeyer flask, 1 L. . Water bath at 70-80°. . Beaker, 50 mL. . Fat extraction apparatus. . Steam bath, . Drying oven. Centrifuge. AAURUL Reagents 1, Ethyl alcohol, 95%. 2. HCI solution. Add 110 mL deionized water to a 1-L Erlenmeyer flask. Slowly add 250 mL concentrated HCI to the flask. Carefully mix and allow to cool to ambient temperature before using. 3. Bthyl ether, free from residue on evaporation. 4, Petroleum ether, boiling point below 60°. Caution. See Note 1. Procedure 1. Place 2 g flour in S0-mL beaker, add 2 mL alcohol, and stir to moisten all particles. (Moistening of sample with alcohol prevents lumping on addition of acid.) Add 10 mL HCI solution, mix well, set beaker in water bath held at 70— 80°, and stir at frequent intervals until sample is completely hydrolyzed (usually 30-40 min. See Note 2. Add 10 mL alcohol and cool. 2. Transfer mixture to fat extraction apparatus. Rinse contents of beaker into extraction tube with 25 mL. ether in three portions. Stopper flask with cork, Neoprene, or other synthetic rubber stopper not affected by solvents, and shake vigorously 1 min. Add 25 ml. redistilled petroleum ether (boiling point below 60°) and again shake vigorously 1 min. 3. Let stand until upper liquid is practically clear, or centrifuge fat extraction flask 20 min at about 600 rpm. 4, Draw off as much ether fat solution as possible through filter consisting of pledget of cotton packed firmly enough in stem of funnel to allow free passage http #felx.doi.org/10.1084/AACCIntMethod-30-10.01Crude Fat AACC International Method Crude Fat in Flour, Bread, and Baked Cereal Products Not Containing Fruit (continued) of ether into weighed 125-mL flask. (Before weighing flask, dry it, and similar flask as counterpoise, in drying oven at 100° and then let stand in air to constant weight). 5. Re-extract liquid remaining in tube twice, each time with only 15 mL of each ether; shake well on addition of each ether. 6. Draw off clear ether solution through filter into same flask as before, and wash tip of spigot, funnel, and end of funnel stem free from suspended water with few mL of mixture of equal volumes of the two ethers. Evaporate ethers slowly on steam bath. 7. Dry fat in drying oven at 100° to constant weight (about 90 min). Remove flask and counterpoise from oven, let stand in air to constant weight (about 30 min), and weigh. (Owing to size of flask and nature of material, there is less error by cooling in air than in desiccator.) 8. Correct this weight by blank determination on reagents used. Calculation weight of fat (corrected for blank) ee eee weight of sample % Fat = 100 Notes 1. Caution. Petroleum and ethyl ether are extremely flammable solvents. Do not let vapors concentrate to flammable level in work area, since it is nearly impossible to eliminate all chance of sparks from static electricity even though electrical equipment is grounded, Use effective fume removal device to remove these vapors when released. Store ethyl ether protected from light. Unstable peroxides can form upon long standing or exposure to sunlight in bottles. These can react explosively with Cl, 03, LiAIE), or strong oxidizing agents. 2. Some produets, such as wheat flour, granolas, and bread products, take up to 90 min to completely hydrolyze. Reference AOAC Intemational. 1995. Official Methods of Analysis of AOAC Intemational, 16th ed. Method 922.06, The Association, Arlington, VA.