High-sensitive LC-MS/MS method for the simultaneous determination of mirodenafil and its major metabolite, SK-3541, in human plasma: Application to microdose clinical trials of mirodenafil

• Published in: Journal of separation science Vol. 36; no. 5; pp. 840 - 848
• Main Authors: Cho, Doo-Yeoun, Bae, Soo Hyeon, Shon, Ji-Hong, Bae, Soo Kyung
• Format: Journal Article
• Language: English
• Published: Weinheim Blackwell Publishing Ltd 01.03.2013; Wiley; Wiley Subscription Services, Inc
• Subjects: Adult; Analysis; Biological and medical sciences; Calibration; Chromatography; Chromatography, High Pressure Liquid - methods; Clinical dosing; Clinical trials; Drug Dosage Calculations; Erectile Dysfunction - drug therapy; Ethers; General pharmacology; High sensitivity; Human; Humans; Male; Mass spectrometers; Mass spectrometry; Mathematical analysis; Medical sciences; Metabolites; Microdosing; Mirodenafil; Monitoring; Pharmacology; Pharmacology. Drug treatments; Plasma; Pyrimidines - blood; Pyrimidinones - blood; Pyrimidinones - metabolism; Pyrimidinones - therapeutic use; Sensitivity and Specificity; Separation; Sulfonamides - blood; Sulfonamides - metabolism; Sulfonamides - therapeutic use; Tandem Mass Spectrometry - methods; Young Adult; Biological fluid; Tandem mass spectrometry; Chemical analysis; 3',5'-Cyclic-GMP phosphodiesterase; Isozyme; Metabolite; HPLC chromatography; Clinical dosing; Esterases; Phosphoric diester hydrolases; Electrospray; Blood; Microdosing; Blood plasma; High sensitivity; Clinical trial; Quantitative analysis; Human; Validation; Healthy subject; Enzyme; Coupled method; Oral administration; Enzyme inhibitor; Phosphodiesterase 5 inhibitor; Sensitivity; Mirodenafil; Simultaneous measurement; Hydrolases; Pharmacokinetics
• ISSN: 1615-9306; 1615-9314; 1615-9314
• DOI: 10.1002/jssc.201200919

A high‐sensitivity LC/MS/MS method was developed and validated for the simultaneous determination of mirodenafil and its major metabolite, SK‐3541, in human plasma. Mirodenafil, SK‐3541, and udenafil as an internal standard were extracted from plasma samples with methyl tert‐butyl ether. Chromatographic separation was performed on a Luna phenyl‐hexyl column (100 × 2.0 mm) with an isocratic mobile phase consisting of 5 mM ammonium formate and ACN (23:77, v/v) at a flow rate of 0.35 mL/min. Detection and quantification were performed using a mass spectrometer in selected reaction monitoring mode with positive ESI at m/z 532.3 → 296.1 for mirodenafil, m/z 488.1 → 296.1 for SK‐3541, and m/z 517.3 → 283.2 for udenafil. The calibration curves were linear over a concentration range of 2–500 pg/mL using 0.5 mL plasma for the microdose of mirodenafil (100 μg). Analytical method validation of the clinical dose (100 mg), with a calibration curve range of 2–500 ng/mL using 0.025‐mL plasma, was also conducted. The other LC‐MS/MS conditions were similar to those used for the microdosing. Each method was applied successfully to pharmacokinetic studies after a microdose or clinical dose of mirodenafil to six healthy Korean male volunteers.