S Vijay Kumar, Dhiman Vinay, Giri Kalpesh Kumar, Sharma Kuldeep, Zainuddin Mohd, Mullangi Ramesh
Drug Metabolism and Pharmacokinetics, Jubilant Biosys Ltd, Industrial Suburb, Yeshwanthpur, Bangalore-560, 022, India.
Biomed Chromatogr. 2015 Sep;29(9):1325-9. doi: 10.1002/bmc.3426. Epub 2015 Jan 26.
A novel, simple, specific, sensitive and reproducible high-performance liquid chromatography (HPLC) assay method has been developed and validated for the estimation of tofacitinib in rat plasma. The bioanalytical procedure involves extraction of tofacitinib and itraconazole (internal standard, IS) from rat plasma with a simple liquid-liquid extraction process. The chromatographic analysis was performed on a Waters Alliance system using a gradient mobile phase conditions at a flow rate of 1.0 mL/min and C18 column maintained at 40 ± 1 °C. The eluate was monitored using an UV detector set at 287 nm. Tofacitinib and IS eluted at 6.5 and 8.3 min, respectively and the total run time was 10 min. Method validation was performed as per US Food and Drug Administration guidelines and the results met the acceptance criteria. The calibration curve was linear over a concentration range of 182-5035 ng/mL (r(2) = 0.995). The intra- and inter-day precisions were in the range of 1.41-11.2 and 3.66-8.81%, respectively, in rat plasma. The validated HPLC method was successfully applied to a pharmacokinetic study in rats.
已开发并验证了一种新颖、简单、特异、灵敏且可重现的高效液相色谱（HPLC）分析方法，用于测定大鼠血浆中的托法替布。该生物分析程序包括通过简单的液-液萃取过程从大鼠血浆中提取托法替布和伊曲康唑（内标，IS）。色谱分析在Waters Alliance系统上进行，使用梯度流动相条件，流速为1.0 mL/min，C18柱保持在40±1°C。使用设置在287 nm的紫外检测器监测洗脱液。托法替布和IS分别在6.5和8.3分钟洗脱，总运行时间为10分钟。按照美国食品药品监督管理局的指南进行方法验证，结果符合验收标准。校准曲线在182-5035 ng/mL的浓度范围内呈线性（r(2)=0.995）。大鼠血浆中的日内和日间精密度分别在1.41-11.2%和3.66-8.81%的范围内。经过验证的HPLC方法成功应用于大鼠的药代动力学研究。
