title: HPLC-UV method development and validation for 16-dehydropregnenolone, a novel oral hypolipidaemic agent, in rat biological matrices for application to pharmacokinetic studies.
abstract: An accurate and precise HPLC assay has been developed and validated for determination of dehydropregnenolone (DHP) in rat plasma, bile, urine and feces. Separation was achieved using a C-18 reversed phase column with a mobile phase comprising of acetonitrile and deionized water (55:45% v/v) using a UV detector, set at a wavelength of 248 nm. The method, applicable to 200-microl plasma, bile and urine, involved double extraction of the samples with n-hexane. The sample clean up for feces involved single extraction of 50 mg of sample with 3 ml of acetonitrile. The method was sensitive with a limit of quantitation of 20 ng/ml in all the matrices and absolute recovery >92%. Precision and accuracy were within the acceptable limits, as indicated by relative standard deviation varying from 4.7 to 11.2% and bias values ranging from 1.8 to 8.8%. Moreover, DHP was stable in plasma, bile and urine up to 90 days of storage at -60 degrees C and after being subjected to three freeze-thaw cycles. The method was applied to generate the pharmacokinetics of DHP in rats after oral and intravenous administration.
authors: Pharmacokinetics and Metabolism Division, Central Drug Research Institute, Chattar Manzil Palace, P.O. Box No. 173, Lucknow 226001, India.
authors: S K
authors: SK
authors: Singh
authors: Nitin
authors: N
authors: Mehrotra
authors: S
authors: S
authors: Sabarinath
authors: R C
authors: RC
authors: Gupta
mesh_terms: D000284:Administration, Oral; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005109:Extracellular Matrix / Q000378:metabolism; D000960:Hypolipidemic Agents / Q000008:administration & dosage / Q000032:analysis* / Q000493:pharmacokinetics*; D008297:Male; D011284:Pregnenolone / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000032:analysis* / Q000493:pharmacokinetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013056:Spectrophotometry, Ultraviolet / Q000379:methods
chemical_list: D000960:Hypolipidemic Agents; C003592:16-dehydropregnenolone; D011284:Pregnenolone