Determination Of Crude Fiber

1. Determination Of Crude Fiber

2. Crude Fiber Definition : A sample is boiled with diluted acid and then with dilute alkali , then washed with ethanol & diethyl ether and the residue is subtracted with its ash . Consist of : Indigestible parts in feed . Lignin , cellulose , hemicellulose, chitin , pentosan .

3. Source : • Crude fiber mainly from : feed of plant origin and small amount from animal origin . • There is a relation ship between plant fiber & nutritional value of the feed . • Feed stuff has high crude fiber , has lower nutritive value .

4. Plant Fiber Consists Of : Cell wall which comprised of lignin , cellulose , hemicelluloses and pectin . To Measure Fiber : Detergent analysis . Enzymatic analysis .

5. 1- Detergent Analysis . Feed is heated using a neutral detergent & separated into soluble and insoluble parts . Organic matter in insoluble part correspond to cell wall and is defined as neutral detergent fiber ( NDF ). Low digestivity in cell wall is measured by heat treatment with an acidic detergent solution and is defined as acid detergent fiber ( ADF ) .

6. 2- Enzymatic Analysis Starch and protein are degraded by enzymes and is separated into soluble and insoluble parts. Organic matter in the insoluble part corresponds to cell wall, and is defined as organic cell wall (OCW). The “indigestible” fraction after the degradation of starch and protein is measured by degrading cellulose with enzymes, and is defined as fiber with low digestivity or organic b fraction (Ob).

8. Crude Fiber Determination 1. Standing method • In a 500-mL tall beaker add 2-5 g of an analysis sample+ 50 mL of sulfuric acid and add water to be 200 mL. • Cover the tall beaker with a condenser. • Boil for 30 minutes while moisture is evaporated . • Add 300 mL of water and leave at rest overnight. • Aspirate the supernatant, add water again to be 200 mL, and subject to the same procedure . • To the residue (acid-insoluble fraction), add 50 mL of sodiumhydroxide solution .

9. Add water to be 200 mL, cover with a condenser . • Filter the residue (acid/alkali-insoluble fraction) with filter paper in an aluminum weighing dish . • Dried at 135±2 °C for 2 hours, cooled in a desiccator, and weighed . • Wash the residue on the filter paper with hot water until the alkaline reaction of filtrate disappears . • Wash 2-3 times with a small amount of ethanol and diethyl ether then air-dry for 3-4 hours.

10. Put the acid/alkali-insoluble fraction with the filter paper in the said weighing dish . • Then dry , cool and weigh . • Calculate the amount of the acid/alkali-insoluble fraction in the sample. • Put in crucible (heated at 550-600 °C for 2 hours, cooled in a desiccator , and accurately weighed . • Subtract the ash content from the amount of the acid/alkali-insoluble fraction to calculate the crude fiber content in the sample.

11. 2-Filtration method • As standing method but instead of over night , filtration with stainless wire mesh or nylon paper .

12. 3-Analysis with automated instruments Fiber test . Fibertec . Fiber analyzer (ANKOM) . Fibretherm FT1 2 .

13. Fiber Analyzer

14. Fiber AnalyzerFilter Paper

15. Fiber Analyzer

16. 4- Heat-stable α-amylase-treated neutral detergent fiber (aNDF and aNDFom) • Neutral detergent fiber (NDF) was developed as an analysis method for total fiber in grass. • Various modification have been done later to be applicable to feed crop . • These aNDF methods use heat-stable α-amylase and sodium sulfite commonly for all feeds.

17. Substances soluble in neutral detergent are digested by bacteria in the digestive organs of livestock. • NDF is the part that is not digestible or is slowly digested by livestock. • Its major components are cellulose, hemicelluloses and lignin.

18. 5- Acid detergent fiber (ADF and ADFom) • The analysis method for acid detergent fiber (ADF) comprehends the fiber fraction in cattle feed . • Measuring fractions with no or low digestivity in fiber. • The major components of ADF are cellulose and lignin.

21. Crude Ash (CA) Definition : • Sample is incinerated without special treatment. • There always is contamination with charred organic matter, resulting in a blackish color.

22. Procedure : • Weigh accurately 2-5 g of an analysis sample and put it in a crucible . • Heated at 550-600 °C for 2 hours. • Cool in a desiccator . • Weigh to calculate the crude ash content in the sample .

23. Container For Incineration : Ceramic. Platinum. Pyrex .

24. Muffle Furnace

25. Nitrogen Free Extract(NFE) Definition : Subtract the contents (%) of the following 5 ingredients: Moisture. Crude protein. Ether extract. Crude fiber . Crude ash. from the whole feed, and the result is shown as: Nitrogen Free Extracts (NFE) (%).

26. Nitrogene Free Extracts Content (%) = • 100 − { moisture content (%) + crude protein content (%) + ether extract content (%) + crude fiber content (%) + crude ash content (%) } The Major Ingredients : • Soluble carbohydrates such as starch and sugars, as well as organic acids. • NFE is an important nutrient as an energy source for animals.

27. Ether Extract (EE) Determination

28. Ether Extract Or Crude Fat Definition : • Dried , ground sample is extracted with diethyl ether which dissolves fat , oil , pigments and other fat soluble substance . • The ether is then evaporated from fat solution . • The result residue is ether extract .

29. Ether extract contain : • Fat , fatty acid , wax , lecithin , cholestrin , phospholipids and oil soluble dyes ( chlorophyll & caotenoids ). • When feeds are stored for along period , EE decrease gradually because unsaturated fatty acids in feed absorb O2 in air and become insoluble in ether .

30. Oil Bearing Seed Seeds that valued mainly for the edible or industrial oils. Assoybeans, coconuts, rapeseed,sunflower . Cereals And Bran • Contain 4 % EE and have high energy value . Oil Meal • Contain 1 % EE and major components are fat and oil. • Have low energy value . Oil cake : The residues obtained after the extraction of oil from oilseed . Oil meal is ground oil cake .

31. Soxhlet Extractor Principle The flask is heated and the solvent evaporates and moves up into the condenser where it is converted into a liquid that trickles into the extraction chamber containing the sample.  When the solvent surrounding the sample exceeds a certain level it overflows and trickles back down into the boiling flask. Fat content is measured by weight loss of the sample or by weight of the fat removed.

34. Procedure Weigh 2-5 gm of sample . Put it in a filter thimble . Dry at 100c for 2hs . Transfer the thimble into a soxhlet extractor . In a flask put diethyl ether and extract for 16hs . Boiling the flask , diethyl ether evaporates and go through tube to condenser which convert vapor to liquid

35. Liquid diethyl descend to the sample present in thimble . This dissolve part of sample which present in thimble and around sample then arrive to the top of siphon . When 2 tube of siphon filled it descend again to the flask . Repeat this cycle .

36. 2-3 drops per second of diethyl ether . 16-20 circulation per hour . After cycles are completed , weigh the flask with dissolved sample and subtracted from empty flask , the result is EE . Or subtract the weight of thimble with sample after the experiment from original weight to calculate EE . Eliminate solvent into roto evaporate or kiln .

38. Notes Drying for long time cause evaporation of lower free fatty acids . In sufficient drying cause oxidation of unsaturated fatty acids . Wipe the bottle with gauze to prevent dust or dirt on hand attach bottle .

39. Automated Instruments Soxtest . Soxtec. Soxtherm . Triglyceride Range of chromatographic range

40. Total fat Continuous extraction . Acid hydrolysis . Mixed solvent extraction . Alkaline hydrolysis. Near infrared reflectance .

41. Fatty acid GLC gas liquid chromatography . HPLC high performance liquid chromatography . Trans fatty acids GLC with infrared analysis . Infrared absorption . GLC.