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.
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.
Zhao
Ying-Yong
YY
Biomedicine Key Laboratory of Shaanxi Province, Northwest University, No.229 Taibai North Road, Xi'an, Shaanxi 710069, China.
Qin
Xiang-Yang
XY
Zhang
Yongmin
Y
Lin
Rui-Chao
RC
Sun
Wen-Ji
WJ
Li
Xiao-Ye
XY
D000818:Animals; D002783:Cholestenones / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D004365:Drugs, Chinese Herbal; D004875:Ergosterol / Q000032:analysis / Q000737:chemistry; D016018:Least-Squares Analysis; D008297:Male; D055492:Polyporus / Q000737:chemistry*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D002783:Cholestenones; D004365:Drugs, Chinese Herbal; C015860:ergosta-4,6,8(14),22-tetraen-3-one; D004875:Ergosterol