Isolation of lipids from tissue

Isolation of lipids from tissue

  • Homogenise 1 g of tissue for 1 minute with 10 ml methanol and add 20 ml chloroform and homogenise well for 2 minutes.
  • Filter the solid and resuspend it in chloroform: methanol of 2:1 v/v 30 ml and homogenise for 3 minutes.
  • Filter and wash the solid again with 30 ml of chloroform and finally with 10 ml methanol.
  • Collect all the filtrate and measure the volume and add 0.88% potassium chloride at a ratio of one quarter of total volume.
  • Mix well and allow settling.
  • Separate the upper layer by aspiration and add water methanol mixture 1:1 at ratio of one quarter of volume of lower layer.
  • Again separate the upper layer and evaporate and dissolve the residue in known volume of solvent and use for spotting TLC plates.
Last modified: Monday, 19 March 2012, 6:27 AM