Thin-layer chromatography with stationary phase gradient as a method for separation of water-soluble vitamins

The group of hydrophilic vitamins play an important role in human health, and their lack or excess produces specific diseases. Therefore, the analysis of these compounds is indispensable for monitoring their content in pharmaceuticals and food in order to prevent some human diseases. TLC was successfully applied in the analysis of hydrophilic vitamins, but the most difficult problem in the simultaneous analysis of all these compounds is to find an optimum stationary phase-mobile phase system due to different chemical characteristics of analytes. Unfortunately structural analogues are difficult to separate in one chromatographic run, and this is the case in hydrophilic vitamins investigations. TLC gives the possibility to perform two-dimensional separations by using stationary phase gradient achieving the highest resolution by combining two systems with different selectivity. The goal of this work was to develop a method of analysis enabling separation of hydrophilic vitamins using TLC with adsorbent gradient. The developed method was used for identifying the water-soluble vitamins in alcoholic extracts of Hippophae rhamnoides and of Ribes nigrum.

► Developed analysis method enables separation of hydrophilic vitamins by TLC-adsorbent gradient.
► Developed method was used for identifying the vitamins in Hippophae rhamnoides and Ribes nigrum.
► Complete separation of the hydrophilic vitamins was possible using silica gel and cellulose plates.
► Spots from first plate can be moved to the second one, without scraping, extraction, and re-spotting.