All solvents were purchased reagent-grade from Roth or Sigma-Aldrich (both Vienna, Austria), unless stated otherwise.
Lipids were analyzed by HPTLC on nano-silica gel glass plates (Sigma-Aldrich) or HPTLC-silica 60 on aluminum sheets (Merck, Darmstadt, Germany) developed in chloroform:methanol:water (65:25:4), referred to here as solvent system A.
For lipid analysis by mass spectrometry (ESI–MS), lipids were either visualized after HPTLC separation under 365-nm UV-light after staining with a primuline (Sigma-Aldrich) solution [18 (link)] and then extracted from the matrix, or purified from the silica plates as described inSection 2.4 .
Other lipid visualization techniques used include the multipurpose Hanessian’s stain for total lipid visualization [19 (link)] and carbohydrate staining with thymol [20 (link)].
Lipids of interest were scratched from the HPTLC plate and the scratched silica matrix was placed in a glass Pasteur pipette packed with glass-wool and the lipids were eluted with chloroform:methanol (1:1). The sample was then filtered through a 0.2-μm PTFE filter (Roth) and dried under a flow of nitrogen.
Lipids were analyzed by HPTLC on nano-silica gel glass plates (Sigma-Aldrich) or HPTLC-silica 60 on aluminum sheets (Merck, Darmstadt, Germany) developed in chloroform:methanol:water (65:25:4), referred to here as solvent system A.
For lipid analysis by mass spectrometry (ESI–MS), lipids were either visualized after HPTLC separation under 365-nm UV-light after staining with a primuline (Sigma-Aldrich) solution [18 (link)] and then extracted from the matrix, or purified from the silica plates as described in
Other lipid visualization techniques used include the multipurpose Hanessian’s stain for total lipid visualization [19 (link)] and carbohydrate staining with thymol [20 (link)].
Lipids of interest were scratched from the HPTLC plate and the scratched silica matrix was placed in a glass Pasteur pipette packed with glass-wool and the lipids were eluted with chloroform:methanol (1:1). The sample was then filtered through a 0.2-μm PTFE filter (Roth) and dried under a flow of nitrogen.