高效液相色谱-质谱联用测定小鼠血浆中FG-4592浓度及其药动学研究

Determination of FG-4592 in mice plasma and pharmacokinetic study by LC-MS/MS method

目的建立LC-MS/MS法检测小鼠血浆内FG-4592的浓度,研究FG-4592在小鼠体内的药代动力学。方法KM小鼠腹腔注射FG-4592 5 mg/kg,不同时间点采集血浆,样品经甲醇沉淀。选用Agilent Zorbax SB-Aq-C18色谱柱（2.1 mm×100 mm,3.5μm）,柱温25℃,流动相为甲醇-水（含5 mmol/L甲酸铵,0.1%甲酸）（90∶10）,流速0.35 m L/min。采用ESI离子源,正离子模式多反应监测,离子通道分别为m/z 353.1→m/z 278.1（FG-4592）和m/z294.9→m/z 235.0（内标物）。结果测定血浆中FG-4592的线性范围为1～1000 ng/m L,日内及日间精密度均〈10%,准确度在90%～105%范围内,方法学考察均符合生物样品的分析要求。药代动力学参数：Cmax为（22.80±0.36）μg/m L,AUC（0-72）为（31.64±0.97）（μg·h）/m L,t1/2为8.81 h。结论本研究建立的FG-4592血药浓度测定方法可行,该药物在血液中吸收良好,测定结果可为药物的基础研究提供依据。

Objective To develop a LC-MS/MS method for the quantitative analysis of FG-4592 in mice plasma and to study the pharmacokinetic profile of FG-4592 with this method. Methods KM mice were intraperitoneally administered5 mg/kg FG-4592. The plasma samples were collected at different time points and precipitated by methanol. An Agilent Zorbax C18（2.1 mm×100 mm, 3.5 μm） chromatographic column with the column temperature of 25℃ was used.The mobile phase consisted of methanol-water（contained 5 mmol/L ammonium formate and 0.1 % formic acid）（90∶10）.The flow rate of mobile phase was 0.35 m L/min. The analyte was monitored by positive electrospray ionization（ESI） in multiple reaction monitoring mode. The mass transition pairs was m/z 353.1→ m/z 278.1（FG-4592） and m/z 294.9→m/z 235.0（internal standard） respectively. Results The calibration curve was established in concentration range of 1-1000 ng/m L. The intra-and inter-day RSDs were both less than 10%. The method recoveries were within 90%-105%and met for the requirements of biological sample analysis. The pharmacokinetic parameters were as follows： Cmaxwas（22.80±0.36） μg/m L, AUC（0-72）was（31.64±0.97）（μg·h）/m L, t1/2was 8.81 h. Conclusion This method is reliable. FG-4592 is well absorbed in mice blood and the results can provide the basis for fundamental research.