Kim Kwang-Youl, Nam Moonsuk, Kwon Hyun-Jung, Kim Kwang-Hyeon, Kang Seung-Hyun, Kim Sang-Il, Kim Cheol-Woo, Cho Sang-Heon
Department of Clinical Pharmacology, Inha University Hospital, Inha University School of Medicine, Incheon 22332, South Korea.
Department of Internal Medicine, Inha University Hospital, Inha University School of Medicine, Incheon 22332, South Korea.
Transl Clin Pharmacol. 2017 Mar;25(1):21-27. doi: 10.12793/tcp.2017.25.1.21. Epub 2017 Mar 15.
A simple, rapid, and reliable UPLC-MS/MS method was developed and validated for the determination of tadalafil in human plasma. The plasma samples were deproteinized with acetonitrile. Chromatographic separation was performed on a Shiseido C18 (100 × 2.1 mm, 2.7 µm) column with isocratic elution using 2.0 mM ammonium acetate and acetonitrile (55:45, v/v) with 0.1% formic acid at a flow rate of 0.7 mL/min. The total run time was 1 min per sample. The quantitative analysis was performed using multiple reaction monitoring at transition of 390.4 → 268.3 for tadalafil and 475.3 → 283.3 for sildenafil as an internal standard. The method was fully validated over a concentration range of 5-1,000 ng/mL with a lower quantification limit of 5 ng/mL. Intra- and inter-day precision (relative standard deviation, %RSD) were within 8.4% and accuracy (relative error, %RE) was lower than -3.2%. The developed and validated method was successfully applied to a pharmacokinetic study of tadalafil (20 mg) in Korean healthy male subjects (n = 12).
建立并验证了一种简单、快速且可靠的超高效液相色谱-串联质谱法（UPLC-MS/MS），用于测定人血浆中的他达拉非。血浆样品用乙腈进行蛋白沉淀。采用岛津C18柱（100×2.1 mm，2.7 µm）进行色谱分离，等度洗脱，流动相为含0.1%甲酸的2.0 mM醋酸铵和乙腈（55:45，v/v），流速为0.7 mL/min。每个样品的总运行时间为1分钟。定量分析采用多反应监测模式，他达拉非的监测离子对为390.4→268.3，西地那非作为内标，监测离子对为475.3→283.3。该方法在5-1000 ng/mL的浓度范围内进行了全面验证，定量下限为5 ng/mL。日内和日间精密度（相对标准偏差，%RSD）均在8.4%以内，准确度（相对误差，%RE）低于-3.2%。所建立并验证的方法成功应用于韩国健康男性受试者（n = 12）中他达拉非（20 mg）的药代动力学研究。
