I am currently studying methods from Folch and I am unable to measure the true water content as I only have a freeze dryer so I can't use the blight method. The folch methodology is a bit confusing for me as this is the first time I am doing lipid extraction.
So here goes for my understanding,
I have to mince up my muscle samples either with LN2 or manual cutting on ice. Then add 2:1 chloroform: methanol, Filter it with normal filter (i only have that to work with)/pipette the clear homogenate, then add chloroform:methanol to final volume of 20X tissue sample. Mix gently and centrifuge and the bottom layer is the crude lipid extract?
Once I remove the top layer then wash it with 0.2vol of water and mix then centrifuge 2-3 times. The bottom layer is my lipid extract.
Am I right?
What is then the calculation to determine the total lipid content of my sample?