大鼠血浆中告达庭的LC-MS-MS测定方法学研究

Study on Determination of Caudatin in Rat Plasma by LC-MS-MS

目的建立大鼠血浆中告达庭浓度的LC-MS-MS测定方法。方法取待测大鼠血浆0.2mL,加入炔诺酮(内标),乙酸乙酯提取、浓缩,取上清液20μL进样分析。流动相为乙腈-水(70∶30),色谱柱为江苏汉邦C18柱(4.6mm×150mm,5μm),流速为1.0mL·min-1,LC-MS-MS多反应离子检测,正离子模式,用于定量分析的离子分别是告达庭:m/z514.1→385.4[M+Na+]和炔诺酮:m/z299.7→108.9[M+H+]。结果告达庭的线性范围为2.5～1000μg.L-1,方法的回收率大于80%,批内和批间的精密度均小于15%,稳定性符合生物样品测定要求。结论该方法经考察符合血浆样品的测定要求,可以应用于血药浓度的测定和药动学研究。

OBJECTIVE To develop a sensitive and specific LC-MS-MS method for the determination of caudatin in rat plasma. METHODS The analyte was extracted from plasma with acetoacetate with norethisterone as internal standard, then separated on a prepacked C18-column. The mobile phase consisted of acetonitrile and water （70： 30） , at a flow rate of 1.0 mL· min^-1. A mass spectrometer was used as detector and operated in the positive ion mode. In multiple reaction monitoring （MRM） mode,the ion combinations of m/z 514. 1→385.4[ M ＋ Na^＋ ] and m/z 299.7→108. 9 [ M ＋ H^＋ ] were used to qualify caudatin and IS,respectively. RESULTS Chromatograms showed no endogenous interfering peaks. Each analysis was completed within 5 min. The caudatin calibration was linear in the concentration range of 2. 5 - 1 000 μg · L^-1. The intra-batch and inter-batch RSDs were less than 15%. The recovery of the extraction was more than 80%. CONCLUSION The method is proved to be suitable for caudatin pharmacokinetics.