Hi Fabiane

Chromatography

Standards (1-20 μL) and samples (1 μL) were applied to the plates using an automated sample applicator Linomat V (Camag, Muttenz, Switzerland) as 3 mm-wide bands 10 mm from the bottom of the plate and 6 mm apart with a delivery rate of 20 s μL-1. Five standards (in duplicate) were applied to the plate alternatively with 14 samples at different concentration. Fermentation samples were indeed deposited at different dilutions to falling into the right concentration range for calibration curves.

You can check in paper:

" Separation and Quantitative Determination of Carbohydrates in Microbial Submerged Cultures Using Different Planar Chromatography Techniques (HPTLC, AMD, OPLC)"

Good Luck

Carbohydrates are difficult to run. They change between open and closed ring structures which elute differently. In the future, please tell us details about your experiment- what sort of TLC (silica, C18, or something else?), the solvents you used, and any modifiers.

Please note a procedure (below) I used for analyzing hydrolyzed carbohydrates below. I needed to see which sugars were in a natural product.

The aqueous extracts were freeze-dried and analyzed for sugars by tlc analysis on a cellulose plate {EM Reagents, Cellulose F-5754, n-BuOH-pyridine-H,O-toluene (10:6:6:1)]. (PDF) Phospholipase D Inhibitors from a Myrsine Species. Available from: https://www.researchgate.net/publication/14525877_Phospholipase_D_Inhibitors_from_a_Myrsine_Species [accessed Aug 06 2020].