Preparative Reversed-Phase High-Performance Thin-Layer Chromatography for Analysis of Anthocyanins

Gradient preparative-layer chromatography and RP-HPTLC have been used for analysis of anthocyanin extracts from Malve arborae L. flos and Vaccinium myrtillus L. fructus. Malvidine was identified in the extract from Malve arborae L. flos. Compounds in the anthocyanin extracts were separated on silica gel 60 F254 PLC plates by five-step gradient elution with mobile phases containing different amounts of methyl tert-butyl ether (MTBE) as modifier. A chamber with a mobile phase reservoir/injector was used in gradient elution. Preparative chromatography experiments were followed by rechromatography on RP-HPTLC plates with a mobile phase gradient prepared from methanol, water, and formic acid. The procedure reported can be used for preparation of anthocyanin standards in the laboratory.