UPLC-MS/MS测定大鼠血浆中8-O-乙酰山栀苷甲酯浓度

Determination the Concentration of Barlerin in Rat Plasma by UPLC-MS/MS

目的建立超高效液相串联质谱法快速测定大鼠体内8-O-乙酰山栀苷甲酯的浓度。方法用乙腈沉淀蛋白方法处理血浆,色谱柱为ACQUITY UPLC BEH C18柱（50 mm×2.1 mm,1.7μm）;流动相为乙腈-0.1%甲酸水,梯度洗脱,流速为0.4 m L·min^-1;用正离子多离子反应监测（MRM）扫描,内标为槲皮素。结果血浆中8-O-乙酰山栀苷甲酯的线性范围为2.5-500 ng·m L^-1（r=0.997 9）,最低定量限为0.5 ng·m L^-1。低、中、高3个浓度（3,45和450 ng·m L^-1）的质控样品的日内、日间精密度RSD均〈10%,3个浓度的相对回收率分别为（103.59±4.75）%,（98.68±4.62）%和（97.06±5.64）%。结论该方法操作简便、快捷,灵敏度高,适于大鼠血浆中8-O-乙酰山栀苷甲酯浓度的测定及其药代动力学研究。

OBJECTIVE To develop an ultra performance liquid chromatography-tandem mass spectrometry method for the determination of barlerin in rat plasma. METHODS Plasmas were precipitated with 200 μL acetonitrile. An ACQUITY UPLC BEH C18（50 mm×2.1 mm, 1.7μm） column was used as the stationary phase. The mobile phase was consisted of acetonitrile and 0.1% formic acid water with gradient elution pumped at a flow rate of 0.4 mL.min^-1. The analytes were detected with positive electrospray ionization in multiple reaction monitoring（MRM） mode and quercetin was used as internal standard. RESULTS Excellent linear calibration curve of barlerin was obtained in the concentration range of 2.5-500 ng-mL^-1 （r：0.997 9）. The lower limit of quantification were 0.5 ng.mL^- 1. The intra and inter day relative standard deviations of low, medium and high quality control samples （3, 45 and 450 ng-mL-1） were all 〈10%. The average recovery were （103.59±4.75）%, （98.68±4.62）% and （97.06±5.64）%. CONCLUSION The method is simple, rapid and sensitive, which is suitable for pharmacokinetics study of barlerin in rats.