HPLC-MS/MS法分析大鼠体内淫羊藿素葡糖醛酸结合物

Analysis of Glucuronide Conjugates of Icaritin in Rat by LC-MS/MS

应用高效液相色谱-串联质谱(HPLC-MS/MS)法鉴别大鼠血浆及尿粪中淫羊藿素葡糖醛酸结合物。生物样品经固相萃取法处理后,以V(乙腈)∶V(水)∶V(乙酸)=45∶55∶1.5为流动相,通过Dikma C18柱分离,采用电喷雾离子四极杆串联质谱在负离子检测方式下进行一级和二级全扫描质谱分析。通过提高离子源内的DP电压,使待测物发生源内裂解产生碎片离子,再对产生的碎片离子进行子离子扫描,可获得相当于三级质谱分析的结果。根据待测物的色谱和质谱信息,发现淫羊藿素在大鼠体内可代谢为1种双葡糖醛酸结合物M1和2种单葡糖醛酸结合物M2、M3。大鼠灌胃给予淫羊藿素后,在血浆和尿样中可检测到M1、M2和M3,在粪便中仅检测到M2和M3。

The glucuronide conjugates of icaritin in rat plasma,urine and feces were analyzed by high performance liquid chromatography-tandem mass spectrometry（HPLC-MS/MS）.Samples were extracted using C18 solid-phase extraction cartridges,and then separated on a Dikma C18 column.The mobile phase was consisted of V（acetonitrile）∶V（water）∶V（acetic acid）= 45∶55∶1.5.Electrospray ionization（ESI） source was applied.Full scan and second stage mass scan were performed in the negative ion mode.According to the chromatography and MS data,a di-glucuronide conjugate（M1） and two mon-glucuronide conjugates（M2 and M3） of icaritin are identified in rat.M1,M2 and M3 can be detected in plasma and urine,while M2 and M3 can be found in feces after oral administration of icaritin to rat.