High performance liquid chromatography coupled to nuclear magnetic resonance spectroscopy and mass spectrometry applied to plant products: Identification of ecdysteroids from Silene otites

HPLC coupled in parallel to nuclear magnetic resonance (NMR) and mass spect rometry (MS) has been used to obtain H-1 NMR and mass spectra of a number o f ecdysteroids present in an extract of the plant Silene otites. Reversed p hase gradient chromatography was performed using a D2O-acetonitrile-based s olvent system. NMR and mass spectra were obtained for integristerone A, 20- hydroxyecdysone, 2-deoxy-20-hydroxyecdysone and 2-deoxyecdysone to provide structural confirmation using continuous and stopped flow HPLC-NMR. The com bined HPLC-NMR-MS system described here provided a more comprehensive analy sis of the ecdysteroids present in the extract than HPLC-NMR alone.