Hello, thank you for your protocol. I performed this analysis recently for the first time; the result was not as expected. And because I have never done a TLC before, I hope you could help me point to where I should troubleshoot my procedure. We don't have the TLC Silica gel 60 F254 as listed in the protocol; so I used the two plates we had to compare. On the left was the Polygram Sil G polyester sheets 20x20 cm (Mfr. No. 805013, Matcherey-Nagel), on the right was a Silica gel on TLC Al foils with fluorescence indicator, 60 Angstrom pore size, 10x20 cm (60800-20EA, Sigma Aldrich). I could see some bands of results mostly from the maltose and maltotriose , but it was not very clear. During the procedure, there was some fluctuation in the oven temperature from 110 to 120 degree Celcius because our oven couldn't hold the temperature very well. Also, at the acid staining step, because we did not have a sprayer, I pour out the staining solution on a glass pane and dip the plates completely, then air dried and put to the oven.

The standards from left to right: D+ galactose, D+glucose, D+xylose, D+maltose, Maltose triose, Maltose monohydrate. On the left plate the last one was all sugars, and the right, just water.

It seems that the TLC Al foil plates worked better, but still the results were not very good. Have you seen this before, and could you give me some suggestions? My email is at trucmai@nmsu.edu.

Thank you very much! 7/23/2020 6:40:42 AM Reply