title: Quantitative analysis of 3alpha,6alpha,24-trihydroxy-24,24-di(trifluoromethyl)-5beta-cholane, a potent synthetic steroidal liver X receptor agonist in plasma using liquid chromatography-tandem mass spectrometry.
abstract: The steroidal liver X receptor agonist, 3alpha,6alpha,24-trihydroxy-24,24-di(trifluoromethyl)-5beta-cholane (ATI-829) is a potential therapeutic agent for the treatment of atherosclerosis. A sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS-MS) method for the quantification of ATI-829 in mouse plasma was developed and validated. Proteins in a 25 microL aliquot of mouse plasma were precipitated, and ATI-829 was extracted from the precipitate by the addition of 125 microL methanol. The overall extraction efficiency was greater than 99%. LC-MS-MS with negative ion electrospray and selected reaction monitoring was used for the quantitative analysis of ATI-829. The lower limit of quantitation of ATI-829 corresponded to 5.0 ng/mL (9.7 nM) plasma. Interference from matrix was negligible. The calibration curve was linear over the range 5-2000 ng/mL. The intra-day precision and inter-day precision of the analyses were <4.5% and <6%, respectively, and the accuracy ranged from 92% to 103%. ATI-829 in plasma was stable for at least 6 h at room temperature, 1 week at 4 degrees C, and 3 weeks at -20 degrees C. The validated method was then utilized for pharmacokinetic studies of ATI-829 administered to mice.
authors.affiliation: University of Illinois College of Pharmacy, Chicago, IL 60612-7231, USA.
authors.forename: Jian
authors.initials: J
authors.lastname: Guo
authors.forename: Dacheng
authors.initials: D
authors.lastname: Peng
authors.forename: Qing
authors.initials: Q
authors.lastname: Dai
authors.forename: Shutsung
authors.initials: S
authors.lastname: Liao
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Wright
authors.forename: Richard B
authors.initials: RB
authors.lastname: van Breemen
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods*; D004355:Drug Stability; D006914:Hydroxysteroids / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D016014:Linear Models; D000071518:Liver X Receptors; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D057093:Orphan Nuclear Receptors / Q000819:agonists; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry
chemical_list: C532886:3,6,24-trihydroxy-24,24-bis(trifluoromethyl)cholane; D006914:Hydroxysteroids; D000071518:Liver X Receptors; D057093:Orphan Nuclear Receptors