TLC was performed by loading samples on silica gel 60F254 plates (Merck, Germany). The loaded samples were developed by a mixture of 95% ethanol/1M acetic acid (5:2, pH 7.5; Kaminski and Eichler, 2014 (link)) and visualized by spraying the plate with 0.5% (w/v) 3,5-dihydroxytoluene in 20% (v/v) sulfuric acid and heating it at 120°C for 5min.
HPLC was performed on an Agilent 1,260 series HPLC system coupled with a UV detector (Agilent Technologies, Inc. United States) using the CarboPac™ PA-100 column (4×250mm, 4μm particle size, Thermo Fisher Scientific, United States). The sample was eluted with a gradient concentration of ammonium acetate, set as 0–30mM (0–12min), 30–60mM (12–22min), 100mM (22–32min), and 0 (32–37min), as the mobile phase at a flow rate of 1.0ml/min and detected at 260nm. The corresponding fractions were combined and concentrated through lyophilization.
HPLC was performed on an Agilent 1,260 series HPLC system coupled with a UV detector (Agilent Technologies, Inc. United States) using the CarboPac™ PA-100 column (4×250mm, 4μm particle size, Thermo Fisher Scientific, United States). The sample was eluted with a gradient concentration of ammonium acetate, set as 0–30mM (0–12min), 30–60mM (12–22min), 100mM (22–32min), and 0 (32–37min), as the mobile phase at a flow rate of 1.0ml/min and detected at 260nm. The corresponding fractions were combined and concentrated through lyophilization.
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