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Analysis of anthraquinones in cell cultures of Cinchona 'Robusta' by HPLC with photodiode array and mass spectrometry detection.

An HPLC-PAD-ESI/MS method has been developed for the analysis of anthraquinones in cell cultures of Cinchona 'Robusta'. Using a C18 column and gradient elution with a mobile phase system containing acetonitrile, water and trifluoroacetic acid, a satisfactory separation of both anthraquinone glycosides and aglycones in a crude dichloromethane extract could be obtained. Robustaquinone B was identified as a major anthraquinone in the extracts, and another five anthraquinones were tentatively identified from spectroscopic data. A number of minor unknown compounds were detected and were distinguished from the known anthraquinones. An isocratic system for the quantitative determination of robustaquinone B has also been developed.