Purification of Phenylalkanoids and monoterpene glycosides from Rhodiola rosea L. roots by high-speed counter-current chromatography

Phytochem Anal. 2013 Feb;24(2):129-34. doi: 10.1002/pca.2391. Epub 2012 Jul 19.


Introduction: Rhodiola rosea L. is a medicinal herb used for its adaptogenic properties. The main active components are the phenylpropanoids collectively referred to as rosavins.

Objectives: To develop an isolation method for phytochemicals present in Rhodiola rosea roots using high-speed counter-current chromatography (HSCCC).

Methodology: The roots of Rhodiola rosea were extracted with methanol and fractionated using liquid-liquid partition and polyamide column clean-up. The purified fraction (100 mg) was subjected to semi-preparative HSCCC using the two-phase solvent system ethyl acetate:butanol:water (3:2:5). The head-to-tail elution mode was employed with a flow rate of 1.5 mL/min and a rotary speed of 1000 rpm.

Results: The separation yielded six main fractions with four components more than 90% pure. The sixth fraction was further purified using semi-preparative HPLC with a Synergi-hydro RP C₁₈ -column to obtain rosin and geranyl 1-O-α-l-arabinopyranosyl(1 → 6)-β-d-glucopyranoside. The main components isolated were rosavin (3.4 mg, 97% purity), salidroside (0.5 mg, 90% purity), benzyl-O-β-d-glucopyranoside (1.2 mg, 85% purity), rosarin (1.3 mg, 99% purity), rosiridin (1.8 mg, 92% purity), rosin (1.2 mg, 95% purity) and geranyl 1-O-α-l-arabinopyranosyl(1 → 6)-β-d-glucopyranoside (6.5 mg, 97% purity). The identity and purity of these components were confirmed using ultrafast liquid chromatography-diode-array detector-MS/MS analysis, ¹H- and ¹³C-NMR spectroscopy.

Conclusion: High-speed counter-current chromatography was successful in the isolation of several phytochemicals present in Rhodiola rosea roots, including two components that are not commercially available.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, High Pressure Liquid
  • Countercurrent Distribution / methods*
  • Disaccharides / chemistry
  • Disaccharides / isolation & purification*
  • Glycosides / chemistry
  • Glycosides / isolation & purification*
  • Magnetic Resonance Spectroscopy
  • Mass Spectrometry
  • Molecular Structure
  • Monoterpenes / chemistry
  • Monoterpenes / isolation & purification*
  • Plant Roots / chemistry*
  • Plants, Medicinal / chemistry
  • Reproducibility of Results
  • Rhodiola / chemistry*


  • Disaccharides
  • Glycosides
  • Monoterpenes
  • rosavin