摘要
A sensitive, rapid, and simple liquid chromatography tandem mass spectrometry(LC-MS/MS) method for simultaneous determination of sunitinib and its active metabolites in plasma of BABL/c nude mice was developed and validated. Plasma samples were pre-treated by protein precipitation with pazopanib and used as an internal standard. Separation was performed on a reversed phase C18 column with a mobile phase composing of 10 m M ammonium formate p H 3.25(adjusted with formic acid) and acetonitrile(65:35, v/v) at a flow rate of 0.5 mL /min. Triple quadrupole tandem mass in multiple reaction monitoring(MRM) mode with electrospray positive ionization was used to monitor all the compounds. The current LC-MS/MS method was highly selective and sensitive with lowest limit of quantitation(LLOQ) of 0.5 ng/mL for both analytes and reliable intra- and inter-day precision and accuracy validated by relative error(RE%) and relative standard deviation(RSD%). Linearity of calibration curve was excellent(r0.99) within a concentration range of 0.5–1000 ng/mL. This method was successfully applied to a pharmacokinetics study on BABL/c nude mice given with single dose of oral administration of sunitinib at 20 mg/kg.
本研究建立并验证了一种灵敏、快速、简单的液质联用方法,用于同时测定BABL/c裸鼠血浆中舒尼替尼及其活性代谢产物SU12662的药物浓度。血浆样品采用蛋白沉淀方法处理,并使用帕唑帕尼作为内标。采用C18反相柱进行分离,流动相为10 mM甲酸胺–乙腈(65:35,v/v,pH 3.25),流速0.5 m L/min。所有化合物均采用电喷雾电离源,正离子方式检测。舒尼替尼及SU12662的最低定量下限均为0.5 ng/m L,线性范围均为0.5–1000 ng/m L(r>0.99)。该方法对舒尼替尼及SU12662的测定均具有良好准确度以及可靠的日内、日间精密度,方法稳定性良好,无明显基质效应。此方法成功用于BABL/c裸鼠口服20 mg/kg舒尼替尼的药物代谢动力学研究。
基金
National Natural Science Foundation of China(NSFC,Grant No.81473277)