高效液相色谱-静电场轨道阱质谱鉴定大鼠血浆中蟾蜍二烯内酯及其代谢物

Identification of Bufadienolides and Its Metabolites in Rat Plasma by High Performance Liquid Chromatography Coupled with Linear Trap Quadrupole-Orbitrap Mass Spectrometry

建立了高效液相色谱-质谱鉴定大鼠口服蟾酥总内酯提取物后血浆中蟾蜍二烯内酯类化合物及其主要代谢物的分析方法。大鼠单剂量灌胃(50 mg/kg)蟾酥总内酯提取物后收集血浆样品,乙腈沉淀蛋白,浓缩定容,采用HPLC-MS分析。利用色谱保留时间、高分辨质谱提供的精确分子量和碎片离子精确质量的信息,鉴定了24种蟾蜍二烯内酯类化合物和20种可能的代谢物。其中5个化合物与相应的标准品一致。本方法快速、灵敏,适用于鉴定血浆中的微量蟾蜍二烯内酯类化合物及其体内代谢物。

A method based on high performance liquid chromatography coupled with Linear trap quadrupole（LTQ）-Orbitrap mass spectrometry（HPLC-LTQ-Orbitrap-MS） was developed for the identification of the bufadienolides and its metabolites in rat plasma.The rat plasmas were collected after oral administration of lactones from ChanSu extract（50 mg/kg）.The plasma samples were precipitated by acetonitrile and concentrated,and then analyzed by HPLC-LTQ-Orbitrap-MS.24 bufadienolides and the 20 metabolites were identified according to their elution orders in C18 column,exact molecular weight and MS/MS fragmentation provided by high-resolution MS/MS.5 standards of bufadienolides were used for corroboration.The method is fast,sensitive and suitable to identify trace bufadienolides and its metabolites in plasma.