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  2. 2. Moisture Determination • • • • • • Weigh 4 – 5 gm of a feed sample in a clean previously dried aluminium basin Place the aluminium basin with lid in the oven at 105 C overnight Cool the basin with lid in a desiccator for 30 min to cool. Weigh the basin. The drying and weighing continues until a constant weight is achieved. Sample may be stored in dessicator for determination of ether extract • %Moisture = (Wt of fresh sample- Wt of dried sample) x 100 Wt of sample taken • %DM = 100 - %Moisture. • Since the water content of feed varied very widely, ingredients and feed are usually compared for their nutrient content on moisture free or dry matter (DM) basis.
  3. 3. Ash Determination • • • • • • • • • • • Place new or clean crucible in muffle furnace. Ignite at 600 for one hr Transfer crucibles from furnace to dessicator Keep in dessicator for 1 hr to cool down at room temperature. Weigh the crucible and record. Add 2-4 g of the sample and weigh again. The crucible is placed into muffle furnace at 600 C for 4 hrs or until whitish-grey ash is obtained. Switch off the muffle furnace Cool down to 200-300 C and transfer to dessicator The crucible is then placed in the desiccator for one hour to cool down to room temperature Weigh the crucibles. %Ash = wt of crucible+ash – wt of crucible x 100 wt of sample
  4. 4. Crude Protein (Kjeldahl method) Digestion: • weigh about 2g of the sample • Transfer the sample into kjeldahl flask • Add 4-5 gm of the catalyst and add 25mls of concentrated sulphuric acid, • Wash the neck of the kjeldahl flask with a jet of water from wash bottle. • Place the flask on digestion assembly and continue heating of flask with occasional turning. • Continue heating for 30 min after solution is clear. • Switch off the heater to cool the flask • Transfer the digest into 100ml volumetric flask. Make up the volume with washing with distilled water.
  5. 5. Distillation: • Steam up the Markham Still apparatus and add about 10ml of the digest into the apparatus via a funnel • Add 10 mls of sodium hydroxide from the measuring cylinder so that ammonia is not lost. • Plug the funnel firmly add a few ml H2O • Distill for 5 min and collect the distillate into a conical flask containg 10 ml of 2% boric acid solutioncontaining screened methyl red indicator. • Collect the droping from the condensor for 1 min • Wash tip of the condenser into the flask
  6. 6. Titration • Alkaline ammonium borate formed is titrated directly with 0.1N HCl. • The titre value (the volume of acid used) is recorded. • The volume of acid used is fitted into the formula which becomes %N = 14 x VA x 0.1 x W x100 1000x100 • VA = volume of acid used W= weight of sample • % Crude Protein = %N x 6.25
  7. 7. Ether Extract • Soxhlet apparatus used for the determination of ether extract has 3 major components – 1. An extractor: comprising the thimble which holds the sample – 2. Condenser: for cooling and condensing the ether vapour – 3. 250ml flask Procedure: • About 150 ml of an anhydrous diethyl ether (petroleum ether) of boiling point of 40-60 C is placed in the flask. • 2-5g of the sample is weighed into a thimble and the thimble is plugged with cotton wool. • The thimble with content is placed into the extractor; the ether is then heated in the flask
  8. 8. Ether Extraction • As the ether vapours condensed to liquid form, drop into the sample in the thimble, • the ether soluble substances are dissolved and are carried into solution through the siphon tube back into the flask. • The extraction continues for at least 4 hrs. The thimble is removed and most of the solvent is distilled from the flask into the extractor. • The flask is then disconnected and placed in an oven at 65 C for 4 hrs, • cooled in desiccator and weighed. • %Ether extract = wt of flask + extract – tare wt of flask x100 • wt of sample
  9. 9. Crude Fibre Estimation • The fat-free material transferred into a flask/beaker and 200mls of pre-heated 1.25% H2SO4 is added and the solution is gently boiled for about 30mins, maintaining constant volume of acid by the addition of hot water. • The buckner flask funnel fitted with whatman filter is preheated by pouring hot water into the funnel. • The boiled acid sample mixture is filtered hot through the funnel under sufficient suction • Wash the residue several times with boiling water (until the residue is neutral to litmus paper) and transfer back into the beaker. • Added 200ml of pre-heated 1.25% Na2SO4 and boil for another 30 mins.
  10. 10. Crude Fibre Estimation • . Filter under suction and wash thoroughly with hot water and twice with ethanol. • The residue dried at 65 C for about 24hrs and weighed. • Transfer the residue into a crucible and place in muffle furnace at 600 C and ash for 4 hrs, • Cool in desiccator and weigh. %Crude fibre = Dry wt of residue before ashing - wt of residue after ashing /wt of sample x100
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