Thin Layer Chromatography-Based Separation of Mycolic Acid Variants: A Method to Separate Mycobacterial Cell Wall Lipids Based on Polarity

To analyze the samples by TLC, cover one wall of the TLC chamber with a piece of filter paper. Add Vaseline on the chamber corners to seal the chamber. Decant the solvent mixture over the filter paper, and add the remaining volume of the solvent to the bottom of the TLC chamber. Close the TLC chamber for at least 20 minutes to saturate it

Meanwhile, dissolve the lipids in the glass tube in 200 to 1,000 microliters of chloroform, and use a capillary glass tube to apply 10 microliters of the lipid-chloroform suspension directly onto the TLC plate. After allowing the sample to dry for five minutes, insert the plate into the saturated TLC chamber, and allow the mobile phase to run through the TLC.

When the solvent reaches 1 centimeter from the upper end of the plate, place the plate under laminar flux until the silica is completely dried. Then, spray the dried plate with 15 to 20 milliliters of 10% Molybdatophosphoric acid hydrate in ethanol, until the plate is bright yellow, and heat the plate for two to five minutes at 120 degrees Celsius.