delta 5,7,9(11)-Cholestatrien-3 beta-ol: a fluorescent cholesterol analogue

Chem Phys Lipids. 1984 Nov;36(1):1-14. doi: 10.1016/0009-3084(84)90086-0.


Structural analysis, a purification scheme and stability information on a fluorescent cholesterol analogue, which has been used as a probe in several model and biological systems, are presented. The proposed structure for the fluorophore, cholestatrien-3 beta-ol, closely resembles that of cholesterol. However, problems of low yield during synthesis and rapid decomposition have impeded its use. This study concerns the synthesis and purification of cholestatrien-3 beta-ol by reverse phase high performance liquid chromatography (HPLC). Unlike cholestatrien-3 beta-ol recrystallized from solvents, the fluorescent sterol purified by HPLC was stable over several months at -70 degrees C either as a white, crystalline powder or in ethanolic solution. In model membranes the fluorescence of cholestatrien-3 beta-ol was stable to ultraviolet (UV) light. A simple spectroscopic assay for purity is presented. Included are detailed absorbance, fluorescence, mass, 1H-NMR, and 13C-NMR spectral analyses. The data confirm the structure of cholestatrien-3 beta-ol proposed, but not proven, over 50 years ago, delta 5,7,9(11)-cholestatrien-3 beta-ol.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Cholestenes / analysis
  • Cholestenes / chemical synthesis*
  • Chromatography, High Pressure Liquid
  • Fluorescent Dyes / chemical synthesis*
  • Magnetic Resonance Spectroscopy
  • Mass Spectrometry
  • Spectrometry, Fluorescence


  • Cholestenes
  • Fluorescent Dyes
  • cholesta-5,7,9-trien-3 beta-ol