HPLC-MS/MS法测定大鼠血浆中酸枣仁皂苷A含量及其药动学研究

Determination of Jujuboside A in Rat Plasma by HPLC-MS/MS and Its Pharmacokinetics Study

建立高效液相色谱-质谱串联(HPLC-MS/MS)法检测大鼠血浆中酸枣仁皂苷A的含量,并应用于药动学研究。大鼠静注(4 mg/kg)或灌胃(20 mg/kg)给药,不同时间取血,血浆样品经处理后,采用HPLC-MS/MS法测定酸枣仁皂苷A含量,色谱柱为Waters YMCTM ODS-AQ S-5 120A(2.0×100 mm)分析柱,以电喷雾离子化串联质谱(ESI)及多反应监测扫描模式(MRM)进行检测,流动相为甲醇-水(0.1%甲酸)=50∶50,流速为0.3 m L/min,柱温为30℃,根据测定结果求算药物的药动学参数。酸枣仁皂苷A在40 ng/m L^4 000 ng/m L的浓度范围内线性关系良好(r2=0.999 1),各浓度水平的精密度及稳定性的RSD均小于15%,提取回收率均大于85%。大鼠注射给药后主要药动学参数分别为Ke0.28/h,t1/22.55 h,AUC0→t2 839.89 h·ng/m L,AUC0→∞3 201.51 h·ng/m L,灌胃给药后主要药动学参数分别为Ke0.51/h,t1/21.35 h,AUC0→t206.02 h·ng/m L,AUC0→∞211.13 h·ng/m L。经计算,酸枣仁皂苷A在大鼠体内的生物利用度为1.32%。本试验所建立的检测方法快速简便、精密度好、灵敏度高及稳定性好,适用于酸枣仁皂苷A在大鼠体内血药浓度测定及药代动力学的研究。

To establish a method for the determination of the concentration of Jujuboside A （JuA） in rat plasma by liquid chromatography-mass/mass spectrometry （HPLC-MS/MS）, and study its pharmacokinetics. The rats were intravenously （4 mg/kg） or oral （20 mg/kg） administrated JuA. Blood samples were drawn from the palpebra at different stages. The concentrations of JuA in the plasma were determined by the HPLC-MS/MS method. Chromatographic separation was carried out with a security guard Waters YMCTM ODS-AQ S-5 120A （2.0 ＆#215;100 mm） （Waters, USA）. HPLC-MS/MS was performed on a triple-quadrupole mass spectrometry equipped with electrospray ionization （ESI） and negative multiple reaction monitoring （MRM）. The mobile phase consisted of methanol-water-formic acid （50∶50∶0.1）, delivered at the flow rate of 0.3 mL/min under 30℃. The method was linear over the concentration range of 40 ng/mL-4 000 ng/mL （r^2=0.999 1）. The relative standard deviations of the assay were less than 15.0%for the low, and the recovery of the method was more than 85 %. The pharmacokinetic parameters were as follows： Ke 0.28/h,t1/2 2.55 h,AUC0→t 2 839.89 h·ng/mL, AUC0→∞3 201.51 h·ng/mL in rat with intravenous administration, and Ke 0.51/h,t1/21.35 h,AUC0→t 206.02 h·ng/mL, AUC0→∞211.13 h·ng/mL in rat with oral administration. The bioavailability of JuA was 1.32%. The developed method was successfully applied to the estimation of the pharmacokinetic parameters of JuA.