for biosurfactants, chloroform: methanol (8:3 v/v) or toluene: ethyl acetate: methanol (6:2:2 v/v/v) and spray with anisaldehyde H2SO4 reagent.

Hi Garima

You have a big bunch of possibilities with TLC, starting with the isolation of your extract, the adsorbent, the solvent mix and the visualization. This is an advantage of TLC comparing to f.e. HPLC.

So first: Sample preparation; you could shrink the number of metabolites that you need to run by using an appropriate extraction solvent or Solid Phase Extraction.

second: usually, your adsorbent is silica and you don' seem to have problems to make your product run and separate there, so there might be no need to change.

Third: Solvent optimization is a huge block. If you have to much spots and want to make every spot moving, it will be difficult to get a propper separation anyway. You might reduce the polarity, adding alkanes f.e. to a value, where more polar components than your product are not eluted and don't run.

Forth: You can focus on functional groups for visualizing, maybe a change to another system will give you chance to pre-exclude un-neccessary metabolites, but i think you have a good provement with anisaldehyd, you could uses Vanillin H2SO4...sometimes helpfull or preview in an Iodine chamber ....unsaturated molecules can fix iodine better than saturated an you can make a staining with starch after.

This link here f.e. is focussing specifically at phospholipids and offers a specific separation for this molecules: http://www.cyberlipid.org/phlipt/pl3a0003.htm

Hi Garima,

Besides what Mr. Frank said above, I think it is also important to consider something else. For example, trail might also be due to high concentration and selection of TLC plate(silica, Al2O3), preparation of TLC plate(alkali treated or sth else).

For developing TLC solvent system, i usually prepare for the reactants first and then change the polarity by adding some other solvent gradually.

Hi Garima

Does "Biosurfactant" means that you look for a glycosilated structure?

if you have sugar(s) inside your molecule, you can use a 5% ethanolic H2SO4 and heat. The carbohydrates will carbonify and many other molecules, like lipids are usually not visualized.

if your matrix is still to complex you can run a 2D-TLC, as described in the link

In addition to what all the others have said, I will also advice you try a multiple development, with the solvent system that showed tailing.

Thank you all for your kind consideration

Do you expect this many products? Perhaps your synthesis needs some optimization. If you have enough materials you could do multiple runs reducing the complexity along the way. Are you planning a prep-TLC. If so whats nice about this method is you can do what I call a continuous Prep-TLC. Leave the top of the chanmber slightly open and choose a single solvent system or a solvent system with similar boiling points for both components. Keep it so your analyte moves slowly and run the TLC overnight.

Good hunting.

Thank you for suggestion, Mr. Peter..

I had the same problem. I tried some solvent systems and one of them showed the best result. You can try the combination of ethyl acetate/ hexane too. I got a good result with 11:1.