Liu Zhenzhen, Zhang Lunhui, Ju Ping, Hou Pengyi, Zhang Yuanyuan, Tang Xing, Bi Kaishun, Chen Xiaohui
School of Pharmacy, Shenyang Pharmaceutical University, Shenyang, China.
School of Pharmacy, Shenyang Pharmaceutical University, Shenyang, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2014 Feb 1;947-948:132-8. doi: 10.1016/j.jchromb.2013.12.029. Epub 2013 Dec 31.
A sensitive and reliable high-performance liquid chromatography-mass spectrometry (LC-MS/MS) method was developed and validated for determination of larotaxel (LTX) and its active metabolites (M1, M2 and M3) in rat plasma. The analytes were extracted by one-step protein precipitation and separated on a Capcell pak C18 column (2.0 mm × 100 mm; 2 μm; Shiseido) using methanol-water as mobile phase at a flow rate of 0.2 mL min(-1) in gradient mode. The method was validated over the concentration range of 2.5-1250 ng mL(-1) for LTX and 1.0-500 ng mL(-1) for M1, respectively, while M2 and M3 were monitored semi-quantitatively and quantified as M1 equivalents. Intra- and inter-day accuracy and precision were within the acceptable limits of less than 15% at all concentrations. Coefficients of correlation (r) for the calibration curves were more than 0.99 for all analytes. The quantitation method was successfully applied for simultaneous estimation of LTX and its metabolites in a pharmacokinetic study after oral administration at different doses of 10, 20, and 40 mg/kg and intravenous administration at the dose 10 mg/kg to Wistar rats, respectively. The results indicated that larotaxel has linear pharmacokinetic characteristics in rats after oral administration and its absolute bioavailability in rats was 12.24%.
建立并验证了一种灵敏可靠的高效液相色谱 - 质谱联用（LC-MS/MS）方法，用于测定大鼠血浆中的拉罗他赛（LTX）及其活性代谢物（M1、M2和M3）。通过一步蛋白沉淀法提取分析物，并在Capcell pak C18柱（2.0 mm×100 mm；2 μm；资生堂）上进行分离，以甲醇 - 水为流动相，流速为0.2 mL min⁻¹，采用梯度模式。该方法分别在2.5 - 1250 ng mL⁻¹（LTX）和1.0 - 500 ng mL⁻¹（M1）的浓度范围内进行了验证，而M2和M3进行半定量监测，并以M1当量进行定量。在所有浓度下，日内和日间的准确度和精密度均在可接受的小于15%的限度内。所有分析物校准曲线的相关系数（r）均大于0.99。该定量方法成功应用于在Wistar大鼠分别口服10、20和40 mg/kg不同剂量以及静脉注射10 mg/kg剂量后的药代动力学研究中同时估算LTX及其代谢物。结果表明，拉罗他赛在大鼠口服后具有线性药代动力学特征，其在大鼠体内的绝对生物利用度为12.24%。
