Thin layer preparation

Separation of lipids

  • Clean the glass plates with acetone and then pour aqueous slurry of gel (20g/plate and 2 ml water per g) into the moat of the spreader and spread the plates with the silica gel at 0.25 m thick.
  • Activate the plates by heating at 100oC for one hour and allow cooling.
  • Then spot out 20-50 microliters of an approximate solution of 1%w/v each lipid in the solvent at above 15cm above bottom.
  • Maintain 2 cm distance between each spot.
  • Mark the solvent front from 1 cm from the top.
  • Place the TLC plates in the TLC chamber with the lower edge immersed in the solvent to a depth of 0.5-1 cm.
  • Keep the plate in the solvent till it reaches the solvent front line.
  • The solvent percolate through the sorbent by capillary action moving the compound to different distance.
  • Remove the plate from the chamber and air dry.
  • Spray with different spray reagents and view the plates under UV light or visible light and locate the lipids.
Last modified: Monday, 19 March 2012, 6:28 AM