采用正负离子切换模式的LC-ESI-MS/MS法测定大鼠灌胃给药后京尼平苷的血浆浓度

摘要

Geniposide is a major bioactive constituent isolated from Gardeniajasminoides Ellis.To evaluate the pharmacokinetics of geniposide in pre-clinical studies,a rapid and specific liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated.After simple protein precipitation,geniposide was analyzed on a Diamonsil（R） C18 column with a mobile phase of 10 mM ammonium acetate and methanol (20:80,v/v) at a flow rate of 0.6 mL/min.Detection was performed in "Truncated" multiple-reaction monitoring (MRM) mode with positive electrospray ionization (ESI) at m/z 411→411 for geniposide,and MRM mode with negative ESI ionization at m/z 415→295 for puerarin (internal standard,IS).Linearity was established in the concentration range from 10.0 to 5000 ng/mL.The extraction recoveries ranged from 84.8％ to 90.5％ at concentrations of 10.0,500 and 4.5× 103 ng/mL.The lower limit of quantification (LLOQ) was 10.0 ng/mL with 50 μL plasma.The validated method was successfully applied to the pharmacokinetic study of geniposide in rats at a dose of 200 mg/kg by oral administration.%建立了一种快速、专属性强的液相色谱-串联质谱方法(LC-MS/MS)测定中药栀子的主要活性成分-京尼平苷的大鼠血药浓度去评价其临床前药动学特征.京尼平苷血浆样品经沉淀蛋白后,采用Diamonsil(R) C18色谱柱进行分析,采用流动相10 mM醋酸铵-甲醇(20∶80,v/v),流速0.6 mL/min.样品测定采用“Truncated”多反应检测模式,采用正离子m/z411→411测定京尼平苷,采用负离子m/z415→295测定内标葛根素.线性浓度范围为10.0-5000 ng/mL,最低定量下限为10.0 ng/mL,样品提取回收率范围为84.8％-90.5％.该确证方法成功应用于大鼠灌胃(给药剂量200 mg/kg)给予京尼平苷后的药动学研究.