Kim Do-Hyung, Cho Ji-Yoon, Chae Soo-In, Kang Bo-Kyung, An Tae-Gil, Shim Wang-Seob, Noh Young Su, Hwang Se Jung, Chung Eun Kyoung, Lee Kyung-Tae
Department of Life and Nanopharmaceutical Sciences, Graduate School, Kyung Hee University, 26 Kyungheedae-ro, Dongdaemun-gu, Seoul 02447, Korea.
Kyung Hee Drug Analysis Center, Kyung Hee University, 26 Kyungheedae-ro, Dongdaemun-gu, Seoul 02447, Korea.
Transl Clin Pharmacol. 2017 Dec;25(4):173-178. doi: 10.12793/tcp.2017.25.4.173. Epub 2017 Dec 20.
We developed a simple, sensitive, and effective ultra-performance liquid chromatography/tandem mass spectrometry (HPLC-MS/MS) method with an electrospray ionization (ESI) interface in multiple reaction monitoring (MRM) and positive ion modes to determine diazepam concentrations in human plasma using voriconazole as an internal standard (IS). Diazepam and IS were detected at transition 285.2→193.1 and 350.2→127.1, respectively. After liquid-liquid extraction (LLE) using 1.2 ml of ethyl acetate:n-hexane (80:20, v/v), diazepam and IS were eluted on a Phenomenex Cadenza CD-C18 column (150 × 3.0 mm, 3 µm) with an isocratic mobile phase (10 mM ammonium acetate in water:methanol [5:95, v/v]) at a flow rate of 0.4 mL/min. The peak retention time was 2.32 min for diazepam and 2.01 min for IS, respectively. The lower limit of quantitation (LLOQ) was 0.5 ng/mL (S/N > 10) using 50 µL of plasma, and no interferences were observed in chromatograms. Our analytical method was fully validated and successfully applied to a bioequivalence study of two formulations of diazepam in healthy Korean volunteers.
我们开发了一种简单、灵敏且有效的超高效液相色谱/串联质谱法（HPLC-MS/MS），该方法采用电喷雾电离（ESI）接口，在多反应监测（MRM）和正离子模式下，以伏立康唑作为内标（IS）来测定人血浆中地西泮的浓度。地西泮和内标分别在跃迁285.2→193.1和350.2→127.1处被检测到。使用1.2 ml乙酸乙酯：正己烷（80:20，v/v）进行液液萃取（LLE）后，地西泮和内标在Phenomenex Cadenza CD-C18柱（150×3.0 mm，3 µm）上，用等度流动相（10 mM乙酸铵水溶液：甲醇[5:95，v/v]）以0.4 mL/min的流速洗脱。地西泮的峰保留时间分别为2.32分钟，内标的峰保留时间为2.01分钟。使用50 µL血浆时，定量下限（LLOQ）为0.5 ng/mL（S/N>10），色谱图中未观察到干扰。我们的分析方法经过了全面验证，并成功应用于健康韩国志愿者中两种地西泮制剂的生物等效性研究。
