title: Quantitative Assay Validation for Oxandrolone in Human Plasma Using LC-MS-MS.
abstract: A high-performance liquid chromatography-tandem mass spectrometry (LC-MS-MS) method for the determination of oxandrolone concentration in human plasma (0.5 mL) was developed and validated according to the 2001 FDA Bioanalytical Guidelines. Oxandrolone is an anabolic steroid used to promote weight gain for cachectic patients with severe burn injuries, HIV/AIDS, hepatitis C and other wasting syndromes. The assay procedure involved a liquid-liquid extraction of oxandrolone and methyltestosterone (the internal standard, IS) from plasma with n-butyl chloride. The organic layer was clarified by centrifugation and evaporated to dryness under a stream of air. The residue was reconstituted in a solution containing 25% methanol and 75% Milli-Q water, and injected onto a Luna C18 reversed-phase HPLC column (30 mm × 2.0 mm, 2 μm). Separation of oxandrolone and methyltestosterone was achieved with a mobile phase starting composition of 55% methanol and 45% ammonium formate buffer at a flow rate of 0.1 mL/min. The total run time was 21 min per sample. Selected reaction monitoring mode was used for quantifying oxandrolone (m/z 307 → 271) and the IS, methyltestosterone (m/z 301 → 149). To the authors' knowledge, this is the first LC-MS-MS method validated for oxandrolone quantification in human plasma. This method can be used in future pharmacokinetic studies involving oxandrolone.
authors.affiliation: Division of Clinical Pharmacology, Department of Pediatrics, University of Utah, 295 Chipeta Way, Suite 1S100, Salt Lake City, UT 84108, USA Department of Pharmaceutics and Pharmaceutical Chemistry, University of Utah, 30 S 2000 E Salt Lake City, UT 84112, USA.
authors.forename: Matthew W
authors.initials: MW
authors.lastname: Linakis
authors.affiliation: Division of Clinical Pharmacology, Department of Pediatrics, University of Utah, 295 Chipeta Way, Suite 1S100, Salt Lake City, UT 84108, USA Department of Pharmacology and Toxicology, University of Utah, 30 S 2000 E Salt Lake City, UT 84112, USA.
authors.forename: Chris
authors.initials: C
authors.lastname: Stockmann
authors.affiliation: Division of Clinical Pharmacology, Department of Pediatrics, University of Utah, 295 Chipeta Way, Suite 1S100, Salt Lake City, UT 84108, USA.
authors.forename: Sarah C
authors.initials: SC
authors.lastname: Campbell
authors.affiliation: Division of Pediatric Cardiology, Department of Pediatrics, University of Utah, 100 N Mario Capecchi Drive, Suite 1500, Salt Lake City, UT 84113, USA.
authors.forename: Richard V
authors.initials: RV
authors.lastname: Williams
authors.affiliation: Division of Cardiothoracic Surgery, Department of Surgery, University of Utah, 100 N Mario Capecchi Drive, Suite 2800, Salt Lake City, UT 84113, USA.
authors.forename: Phillip T
authors.initials: PT
authors.lastname: Burch
authors.affiliation: Division of Cardiothoracic Surgery, Department of Surgery, University of Utah, 100 N Mario Capecchi Drive, Suite 2800, Salt Lake City, UT 84113, USA.
authors.forename: Linda M
authors.initials: LM
authors.lastname: Lambert
authors.affiliation: Division of Clinical Pharmacology, Department of Pediatrics, University of Utah, 295 Chipeta Way, Suite 1S100, Salt Lake City, UT 84108, USA.
authors.forename: Catherine M T
authors.initials: CM
authors.lastname: Sherwin
authors.affiliation: Department of Pharmacology and Toxicology, University of Utah, 30 S 2000 E Salt Lake City, UT 84112, USA.
authors.forename: Christopher A
authors.initials: CA
authors.lastname: Reilly
authors.affiliation: Division of Clinical Pharmacology, Department of Pediatrics, University of Utah, 295 Chipeta Way, Suite 1S100, Salt Lake City, UT 84108, USA michael.spigarelli@hsc.utah.edu.
authors.forename: Michael G
authors.initials: MG
authors.lastname: Spigarelli
mesh_terms: D045930:Anabolic Agents / Q000097:blood*; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000592:standards; D004355:Drug Stability; D005260:Female; D006801:Humans; D008297:Male; D010074:Oxandrolone / Q000097:blood*; D011237:Predictive Value of Tests; D012015:Reference Standards; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000592:standards
chemical_list: D045930:Anabolic Agents; D010074:Oxandrolone