Plates silica gel 60f254

Recoated aluminum plates silica gel 60F₂₅₄ (E. Merck, Darmstadt, Germany) for TLC were used and visualized under UV at 254 and 366 nm and by spraying with anisaldehyde sulfuric and FeCl₃ reagents. Column silica gel 70–230 mesh and flash silica gel 230–400 mesh (E. Merck, Darmstadt, Germany) were used for the column chromatography. Melting points were determined on a Micro-Química APF-300 apparatus (Micro-Química, Florianópolis, SC, Brazil) and are uncorrected. The purity of the isolated compound was examined by thin layer chromatography (TLC) using Merck silica gel precoated aluminum plates (thickness = 200 μm) and several solvent systems of different polarities. All the reagents and solvents used were of analytical grade and were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA) and Merck KGaA (Darmstadt, Germany).

Silica gel 60 Å grade (particle sizes 15–40 μm and 40–63 μm) for column chromatography was purchased from Merck (Darmstadt, Germany). Sephadex LH-20 beads (size 25–100 μm) were purchased from Sigma-Aldrich (St Louis, MO). Thin-layer chromatography (TLC) plates (silica gel 60 F254, thickness 0.2 mm) were obtained from Merck. Chemical spots on TLC plates after development were detected using p-anisaldehyde-sulfuric acid (AS) and ferric chloride staining reagents, together with I₂ vapor. All solvents were distillated and purified before use.