Korea Institute of Toxicology, Yuseong, Daejeon 305-343, Republic of Korea.
J Pharm Biomed Anal. 2012 Nov;70:587-91. doi: 10.1016/j.jpba.2012.05.030. Epub 2012 Jun 4.
FK-3000 can inhibit proliferation of carcinomas and arrest the growth of carcinoma cells through cytotoxic (apoptosis induction) and cytostatic (cell cycle arrest) effects. A rapid and sensitive assay was developed and validated using liquid chromatography-mass spectrometry (LC-MS) for FK-3000 in rat plasma. FK-3000 was extracted with ethyl acetate from rat plasma samples, and the residue containing the FK-3000 was dried in a gentle stream of nitrogen and reconstituted with acetonitrile. The FK-3000 was quantified using high-performance liquid chromatography (HPLC; Waters Alliance 2695) with a reversed phase Gemini column (3 mm × 150 mm, 5 μm; Phenomenex, USA) and a Waters Micromass ZQ detector. FK-3000 and phenazine, an internal standard (IS), were analyzed by selected ion monitoring (SIM) at m/z transitions of 418.45 and 256, respectively. A lower limit of quantification (LLOQ) of 10 ng/mL was observed, with a linear dynamic range from 10 to 10,000 ng/mL (R>0.999). The accuracy, precision, recovery, matrix effects, and stability of the assay were deemed acceptable according to the FDA guidance for industry (bioanalytical method validation). The FK-3000 concentration was measured in plasma samples up to 6 h following FK-3000 administration at an oral dose of 20 mg/kg. The findings indicate that the assay method is suitable for routine pharmacokinetic (PK) studies of FK-3000 in rats.
FK-3000 通过细胞毒性（诱导细胞凋亡）和细胞抑制（细胞周期阻滞）作用抑制癌细胞的增殖并阻止癌细胞生长。使用液相色谱-质谱联用（LC-MS）法建立并验证了一种快速灵敏的 FK-3000 检测方法，用于检测大鼠血浆中的 FK-3000。FK-3000 从大鼠血浆样品中用乙酸乙酯提取，含有 FK-3000 的残留物在氮气的温和气流中干燥并用乙腈重新配制。FK-3000 使用高效液相色谱（HPLC；Waters Alliance 2695）进行定量分析，采用 Gemini 反相柱（3mm×150mm，5μm；Phenomenex，美国）和 Waters Micromass ZQ 检测器。FK-3000 和吩嗪（内标，IS）通过选择离子监测（SIM）在 m/z 转换为 418.45 和 256 时进行分析。观察到 10ng/mL 的定量下限（LLOQ），线性动态范围为 10 至 10,000ng/mL（R>0.999）。根据美国 FDA 工业指南（生物分析方法验证），该测定法的准确性、精密度、回收率、基质效应和稳定性被认为是可接受的。在以 20mg/kg 的口服剂量给予 FK-3000 后，可在 6 小时内检测到 FK-3000 给药后血浆样品中的 FK-3000 浓度。研究结果表明，该测定方法适用于大鼠中 FK-3000 的常规药代动力学（PK）研究。
