Quantitative HPLC method and pharmacokinetic studies of ergosta-4,6,8(14),22-tetraen-3-one, a natural product with diuretic activity from Polyporus umbellatus

Biomed Chromatogr. 2010 Oct;24(10):1120-4. doi: 10.1002/bmc.1414.


A simple and specific HPLC method with dual wavelength UV detection for the determination of ergosta-4,6,8(14),22-tetraen-3-one (ergone) in rat plasma was developed and proved to be efficient. The method used ergosterol as internal standard (IS). Following a single-step protein precipitation, the analyte and IS were separated on an Inertsil ODS-3 column with a mobile phase containing methanol-water (99:1, v/v) at a flow rate of 1 mL/min. The analytes were detected by using UV detection at wavelength of 350 (ergone) and 283 (IS) nm, respectively. The calibration curve was linear over the range of 0.1-2.0 µg/mL and the lower limit of quantification was 0.1 µg/mL. The intra-day and inter-day precision studies showed good reproducibility with RSD less than 8.5%. The intra-day and inter-day accuracy ranged from 95.6 to 104%. Mean extraction recovery was above 95% at the low, medium and high concentrations. The present HPLC-UV method was simple and reliable. The method described herein had been successfully applied for the pharmacokinetic studies in male SD rats after administration of 20 mg/kg dose of solution of ergone.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cholestenones / blood*
  • Cholestenones / chemistry
  • Cholestenones / pharmacokinetics
  • Chromatography, High Pressure Liquid / methods*
  • Drug Stability
  • Drugs, Chinese Herbal
  • Ergosterol / analysis
  • Ergosterol / chemistry
  • Least-Squares Analysis
  • Male
  • Polyporus / chemistry*
  • Rats
  • Rats, Sprague-Dawley
  • Reproducibility of Results
  • Sensitivity and Specificity


  • Cholestenones
  • Drugs, Chinese Herbal
  • ergosta-4,6,8(14),22-tetraen-3-one
  • Ergosterol