LC-MS/MS法同时测定Beagle犬血浆中柚皮苷和新橙皮苷的含量

LC-MS/MS Simultaneous Determination of Naringin and Neohesperidin in Beagle Dog Plasma

目的:建立同时测定Beagle犬血浆中柚皮苷和新橙皮苷的LC-MS/MS分析方法。方法:以芦丁为内标,血浆样品经乙腈沉淀蛋白处理,采用Diamonsil C18(150 mm×4.6 mm,5μm)色谱柱,以乙腈-0.1%甲酸水(25:75)为流动相,流速为0.4 ml/min,柱温20℃;采用ESI(+)离子源,MRM扫描检测离子对为柚皮苷m/z 581→273、新橙皮苷m/z 611→303、芦丁(内标)m/z611→303。结果:血浆中柚皮苷在0.025～2.5μg/ml、新橙皮苷在0.015～1.5μg/ml浓度范围内线性关系良好,二者的方法回收率均近100%,分析方法的日内日间精密度RSD值均小于10%,内源性物质不干扰样品测定。结论:本方法简便、准确、灵敏度高、专属性好,适用于Beagle犬血浆中柚皮苷和新橙皮苷测定及其体内药代动力学的研究。

Objective： To establish an LC-MS/MS method for the simultaneous determination of naringin and neohesperidin in Beagle dog plasma.Method： The plasma protein was precipitated by acetonitrile.The Diamonsil Cl8（150 mm×4.6 mm,5 μm）column was used as analytical column with a mobile phase consisted of acetonitrile-0.1%formic acid（25：75）.The flow rate was 0.4 ml/min.The temperature of column was 20℃.Rutin was adopted as the internal standard.The detection was performed by MRM mode via electrospray ionization（ESI）sourse operating in the positive ionization mode;The precursor-to-product ion transitions of the analyte naringin is m/z 581→273,neohesperidin is m/z 611→303,rutin（internal standard）is m/z 611→303.Results： The method had good linear relationship within the range of 0.025-2.5 μg/ml for naringin and 0.015-1.5 μg/ml for neohesperidin in plasma.The method recovery of two drugs is nearly 100% and the RSD for inter-and intra-day were less than 10%.Endogenous matrix did not interfere with the analysis.Conclusions： The method was shown to be simple,accurate,sensitive and good reproducibility for the determination of naringin and neohesperidin in beagle dog plasma.It was suitable for pharmacokinetics study of naringin and neohesperidin.