HPLC analysis of anthraquinone derivatives in madder root (Rubin tinctorum) and its cell cultures

A sensitive and reproducible RP-HPLC method was developed for the characterization of madder root and its cell cultures extracts and for the determination of anthraquinone derivatives as glycosides (ruberythric acid, lucidin primveroside) and aglycones (alizarin, lucidin, purpurin) in them. The fingerprint chromatographic patterns of the extracts were obtained on C-18 silica column, using gradient with acetonitrile-water-acidic buffer eluent system. For quantitative measurements, the anthraquinone derivatives were separated by isocratic elution in the same type of mobile phase. The chief components were identified and quantitatively determined. The natural plant and its cell suspension cultures were compared to each other. The preparative fractionation of the extracts was achieved by gel chromatography, HPLC and selective extractions with solvent series, solid-phase extraction techniques and SFE.