HPLC-MS法测定大鼠血浆中艾芬地尔浓度及药代动力学研究

目的: 建立HPLC-MS法测定大鼠体内艾芬地尔的血药浓度,并研究酒石酸艾芬地尔在健康SD大鼠体内的药代动力学特征.方法: 血浆样品碱化后采用乙酸乙酯液-液萃取处理,酮康唑为内标.采用C₁₈色谱柱(5 μ m,4.6 mm×150 mm),以甲醇-pH 7.20醋酸铵溶液为流动相,梯度洗脱(0~3 min,20%A→90%A,80%B→10%B),流速0.6 mL·min^-1.质谱检测采用电喷雾(ESI)离子源,正离子模式,采用选择离子检测(SIM)方式检测,检测离子分别为m/z 326.3(艾芬地尔)、m/z 531.0(酮康唑).选取健康SD大鼠[(250±20)g,雌雄各半],腹腔注射酒石酸艾芬地尔1 mg·kg^-1后,采用建立的HPLC-MS法测定各个时间点(0、10、20、30、40、50、60、75、90、100、110、120、150、180、240、360 min,每个时间点6只)大鼠血浆中艾芬地尔的浓度,并计算其药代动力学参数.结果: 艾芬地尔浓度在0.5~120 μg·L^-1范围内,线性良好(r= 0.9974);日间、日内精密度RSD均小于10.5%,提取回收率均高于88%,专属性良好,血浆样品在本实验的条件下稳定,空白基质中的内源性物质不干扰待测组分和内标的测定.结论: 本分析方法对血浆中艾芬地尔的检测具有专属性,且灵敏、可靠.酒石酸艾芬地尔经腹腔注射后在大鼠体内药动学模型为单室模型,其达峰浓度为113.79 μg·L^-1,半衰期为2.42 h.

Objective: To develop an HPLC-MS method for the determination of ifenprodil in rat plasma and study its pharmacokinetic characteristics in healthy SD rats. Methods: Liquid-liquid extraction of ifenprodil from plasma samples with ethyl acetate was conducted after alkalization by ammonia water.Ketoconazole was used as the internal standard.The C₁₈ column(5 μ m, 4.6 mm×150 mm)was used with the mobile phase of methanol-ammonium acetate solution(pH 7.20)and gradient elution(0-3 min, 20%A→90%A, 80%B→10%B)at the flow rate of 0.6 mL·min^-1.The detection was performed in single ion monitor in positive ionization mode with an electrospray ionization(ESI)source.The single ion monitored were respectively m/z 326.3 for ifenprodil and m/z 531.0 for ketoconazole.Healthy SD rats [(250±20)g, half male and half female] received intraperitoneal injection of ifenprodil tartrate(1 mg·kg^-1).The concentration and the pharmacokinetic parameters of ifenprodil were calculated by HPLC-MS for every six healthy rats at each time point(0 min, 10 min, 20 min, 30 min, 40 min, 50 min, 60 min, 75 min, 90 min, 100 min, 110 min, 120 min, 150 min, 180 min, 240 min, 360 min)after administration. Results: The assay was linear in the range of 0.5-120 μg·L^-1 .The concentrations with a correlation coefficient(r)of 0.9974.RSDs of the intra-day and inter-day precision tests were lower than 10.5% and the extraction recovery was higher than 88%.The stability of this HPLC-MS method was satisfactory.There was no obvious interference of endogenous substances in black matrix on ion suppression or enhancement phenomenon. Conclusion: The analytical method developed is sensitive, specific, rapid, and reproducible, and it is suitable for the pharmacokinetic study of ifenprodil in rats.The peak concentration of ifenprodil tartrate in rats is 113.79 μg·L^-1 and the half-life is 2.42 h.The pharmacokinetic model after intraperitoneal injection of ifenprodil tartrate in rats is a single-compartment model.