nto the fat extraction flask for analysis. Firstly, 1.5 ml of
ammonium hydroxide was added and mixed followed by
10 ml of alcohol (95 %) and the contents were again well
mixed. Secondly, 25 ml diethyl ether was added to the flask,
then it was shook vigorously for 1 min. Finally, 25 ml of light
petroleum ether (b.p. 40–60 °C) was added and the flask was
shook vigorously for 1 min. After separation was complete,
the fat solution was transferred into a petri dish and the petri
dish was dried at 102±2 °C for 1 h and weighed. The total fat
content was calculated as the difference between weight of
petri dish with fat and weight of initial petri dish.