Effective two-dimensional counter-current chromatographic method for simultaneous isolation and purification of oridonin and ponicidin from the crude extract of Rabdosia rubescens

J Chromatogr A. 2007 Mar 30;1146(1):125-30. doi: 10.1016/j.chroma.2007.01.141. Epub 2007 Feb 16.

Abstract

A preparative two-dimensional counter-current chromatographic system (2D-CCC) for simultaneous separation and purification of oridonin and ponicidin from the crude extract of Rabdosia rubescens was presented. It was based on the use of a high-speed CCC (HSCCC, total column volume V(c)=146ml) instrument in the first dimension (1st-D) and a preparative upright CCC (UCCC, V(c)=1500ml) column in the second dimension (2nd-D). The interface was a homemade column-switching system with a 50-ml sample loop. In this way, almost the whole interested region from the first dimension was transferred on-line to the second dimension for further separation. The use of a pair of two-phase solvent systems composed of n-hexane-ethyl acetate-methanol-water (1:5:1:5 and 3:5:3:5, v/v) in the two dimensions permitted the simultaneous separation of oridonin and ponicidin, making this 2D-CCC system with satisfactory resolution and peak capacity. During about 9h separation, two injections with about a 250mg amount of the crude extract for each, was accomplished using this 2D-CCC system, yielding 60mg of oridonin (1) and 10mg of ponicidin (2) all at a purity of over 95% as determined by HPLC analysis. Their chemical structures were identified by electrospray ionization (ESI) MS, (1)H NMR and (13)C NMR.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, High Pressure Liquid
  • Countercurrent Distribution / instrumentation
  • Countercurrent Distribution / methods*
  • Diterpenes / analysis*
  • Diterpenes / chemistry
  • Diterpenes / isolation & purification
  • Diterpenes, Kaurane / analysis*
  • Diterpenes, Kaurane / chemistry
  • Diterpenes, Kaurane / isolation & purification
  • Isodon / chemistry*
  • Magnetic Resonance Spectroscopy
  • Molecular Structure
  • Plant Extracts / analysis
  • Plant Extracts / chemistry
  • Plant Extracts / isolation & purification
  • Reproducibility of Results
  • Spectrometry, Mass, Electrospray Ionization

Substances

  • Diterpenes
  • Diterpenes, Kaurane
  • Plant Extracts
  • oridonin
  • ponicidin