Abstract:

The methods using silica column chromatography, reversed-phase high performance liquid chromatography, liquid chromatography-electrospray ionization mass spectrometry and nuclear magnetic resonance (RP-HPLC/LC-ESI-MS/NMR) for the isolation, purification and identification of geniposidic acid (GPA) in Eucommia ulmoides Oliv. were established. The crude extract of E. ulmoides was obtained after E. ulmoides was extracted with 70% ethanol and the extract was concentrated under reduced pressures. The obtained crude extract dissolved in methanol was injected to a silica gel column and eluted with chloroform-methanol. The eluted fractions were analyzed by RP-HPLC. The eluant from chloroform-methanol (8∶1, v/v) was indicated to be a single component by HPLC with a retention time of 5.142 min. The HPLC analysis of the mixture of this fraction with a GPA standard resulted in a single peak. The ultraviolet and infrared spectra of the eluted solution were essentially the same to those of GPA. On the other hand, LC-ESI-MS, 1H- and 13C-NMR experiments also indicated that the fraction was identical to GPA.

Key words: Eucommia ulmoides Oliv.； traditional Chinese medicine , geniposidic acid, liquid chromatography-electrospray ionization mass spectrometry, nuclear magnetic resonance, reversed-phase high performance liquid chromatography, column chromatography