Bligh and Dyer method
Methanol+chloroform (1:2) (40 mL) mixture was added to the algae sample
and homogenized. Then, 20 mL of 0.4% CaCl2 solution was
added onto these samples. The samples filtered on a filter paper and
then filtered into the tared balloons kept in the oven at 105 °C for 2
hours. These balloons were kept in a dark environment overnight and the
upper layer consisting of methanol+water was separated with the help of
a separation funnel. Chloroform from the chloroform + lipid part in the
solution remaining in the flask was evaporated using a rotary evaporator
(Heidolph) with the help of a water bath at 60 ºC.