HPLC-MS法测定虎杖活性成分在大鼠体内的药代动力学

目的:建立一种有效的高效液相色谱-质谱联用(HPLC-MS)的分析方法,以测定虎杖活性成分白藜芦醇和虎杖苷在大鼠血浆中的药代动力学。方法:以染料木素作为内标物。采用Shim-pack VP-ODS C₁₈分析柱(250 mm×4.6 mm,5 μm)分离。色谱条件:柱温设定为30 ℃;线性梯度洗脱:A相为0.05%甲酸,B相为乙腈,起始时B相20%,5 min后B相100%,维持10 min;流速为1.0 mL·min^-1,通过分流器将0.2 mL·min^-1的流出液注入离子源。质谱条件:选择监测离子检测(SIM):白藜芦醇m/z 229,虎杖苷m/z 391,内标染料木素 m/z 271。给大鼠分别灌胃白藜芦醇、虎杖苷、白藜芦醇+虎杖苷混合溶液和虎杖提取物灌胃液,测定大鼠血浆中的白藜芦醇和虎杖苷浓度。结果:根据所测得的白藜芦醇和虎杖苷在大鼠体内的药动学参数,可知白藜芦醇及虎杖苷在大鼠中的药动学模型均符合二室开放模型。虎杖苷进入大鼠体内后可能被迅速代谢为其苷元——白藜芦醇。

Objective:To develop a specific and rapid method for the plasma pharmacokinetics investigation of resveratrol and polydatin using high performance liquid chromatography coupled with mass spectrometry(HPLC-MS).Methods:Genistein was used as an internal standard(IS).Separation was performed on a Shim-pack VP-ODS C₁₈ column(250 mm×4.6 mm,5 μm)with column temperature set to 30 ℃.Through a solvent splitter,a portion of 0.2 mL·min^-1 was delivered into the ion source.A quadrupole mass spectrometer operated under selected ion monitoring(SIM)mode was used for the determination.A linear gradient elution was selected at the flow rate of 1.0 mL·min^-1:phase A was 0.05% formic acid,phase B was acetonitrile.At the start phase B was set to 20%,altered to 100% after 5 minutes and maintained 10 minutes.The concentration of resveratrol and polydatin in animals was determined after stomach-lavaging with resveratrol,polydatin,resveratrol+polydatin mixture and Rhizoma Polygoni Cuspidati extract intragastric solutions,respectively.Results:The pharmacokinetic data of resveratrol and polydatin fitted well to a two-compartment model with first-order absorption,polydatin rapidly metabolized to its aglycone resveratrol by liver drug metabolism CYP 450 enzymes in liver and/or intestinal flora in vivo.Conclusion:The HPLC-MS method is fast,simple and accurate,especially for the determination of medicine ingredients in blood,which can be an effective method for traditional Chinese medicine composition determination in vivo and deserves further study.