ROSE-GOTTLIEB METHOD

1. SCOPE
- This method is applicable to determine the crude fat in extruded feed which needs pre-
hydrolysis
2. PURPOSE AND SIGNIFICANCE
- After high temperature, shear force and pressure, proportion of combined fat increases. As
a result, this portion of fat could not be extracted with ether. Therefore, pre-hydrolysis is
needed to determine the crude fat content accurately.
3. PRINCIPLE
- The Roese-Gottlieb method is based on the extraction of fat with a mixture of ethyl ether
and petroleum ether in the presence of a concentrated solution of ammonia and ethyl
alcohol. The ammonia solution dissolves the proteins, ethyl alcohol breaks the emulsion and
combination of fats and proteins, avoiding the formation of a gelatinous mixture, the ethyl
ether acts as a solvent for the fat, but to prevent it from also dissolving other components
present in the aqueous phase, it is combined with petroleum ether to reduce the solubility
of the aqueous phase in the ethyl ether. Finally, the fat is extracted, dried and subsequently
determined by weighing.
4. APPARATUS & REAGENTS
- Grinder
- Analytical Balance
- Oven
- Special Extraction Tubes
- NH4OH
- Petroleum Ether
- Ethyl Ether
- 95% EtOH
5. PROCEDURE
1. Weigh accurately 1-2g of sample into a 50 mL beaker and dissolve carefully in warm distilled
water (do not allow to form lumps) with the help of stirring rod
2. Add 1 mL of 0.88(spe.gravity) ammonia and mix well
3. Add 10 mL of ethanol, mix and cool
4. Transfer the mixture into a 100 mL separatory funnel with the help of 10-15 mL distilled
water. Add 25 mL of diethyl ether, stopper the funnel and shake vigorously for one minute
5. Wait for few minutes and add 25 mL of petroleum ether, shake vigorously for 30 seconds
6. Stand for 30 minutes, until two layers get separated.
7. If the two layers get separated, drain the ether layer into a conical flask
8. Repeat the extraction twice using 25 mL portions of diethyl ether and petroleum ether (125
mL each)
9. Combine all three ether extracts in a conical flask, dry over anhydrous MgSO4, and filter into
a previously weighed, clean, dry evaporating dish
10. Evaporate off the solvent on a warm bath and dry over an oven until you get two
consecutive readings of the same value.
6. CALCULATION
Fat percent w/w = Weight of fat  x 100/Weight of Sample