HPLC法测定大鼠血浆中咪达唑仑的含量及其在药物相互作用研究中的应用  

HPLC determination of midazolam in rat plasma and applied to investigation the drug interaction

作　　者：赵娜萍[1] 余露山[1] 曾苏[1] 

机构地区：[1]浙江大学药学院药物分析与药物代谢研究室,杭州310031

出　　处：《药物分析杂志》2007年第6期821-824,共4页Chinese Journal of Pharmaceutical Analysis

基　　金：国家自然科学基金(03225047;30600772)

摘　　要：目的:建立高效液相色谱法测定大鼠血浆中咪达唑仑含量,并应用于佐米曲坦对大鼠肝细胞中 CYP3A 的诱导作用的研究。方法:采用色谱柱为 Diamonsil^(TM)(钻石)C_(18)(200 mm×4.6 mm,5μm),流动相为乙腈-0.01 mol·L^(-1)磷酸二氢钾缓冲液(pH 5.0)(58:42),流速为1.0 mL·min^(-1),检测波长为220 nm。咪达唑仑(10 mg·kg^(-1))作为经典 CYP3A 探针底物对3组不同诱导后的大鼠(佐米曲坦样品组、地塞米松阳性对照组或空白对照组)静注给药,测定咪达唑仑的清除率等参数来研究CYP3A 活性的变化。结果:咪达唑仑在0.3～375 μmol·L^(-1)浓度范围内呈良好的线性关系(r=0.9997,n=5),检测限为0.0018 μmol·L^(-1),平均回收率为97.9%。经佐米曲坦诱导的雄性大鼠组中咪达唑仑 AUC_(0-t)和 T_(1/2)与雄性空白大鼠组中咪达唑仑 AUC_(0-t)和 T_(1/2)比,显著降低,清除率增加,与地塞米松组中咪达唑仑的动力学参数相近;而经佐米曲坦诱导的雌性大鼠的动力学参数与空白雌性组比没有明显的变化。结论:本文建立的大鼠血浆中咪达唑仑测定方法稳定、准确、简便,能使咪达唑仑作为探针药物很好地运用于佐米曲坦对大鼠肝中 CYP3A 诱导作用研究中。另外佐米曲坦对大鼠肝中 CYP3A 具有性别差异的诱导作用。Objective ： To develop an RP - HPLC method for the determination of midazolam in rats plasma. And the method was applied to investigate the induction effect of zolmitriptan on rat hepatic CYP3A activity in vivo. Method： HPLC for midazolam ： The analytical column was DiamonsilTM C18 （200 mm × 4. 6 mm, 5 μm） , the mobile phase consisted of acetonitrile - 0. 01 mol · L^- 1 potassium dihydrogen phosphate buffer solution （ pH 5.0 ） （ 58： 42 ）, the flow rate was 1.0 mL · min - 1, and detected at 220 nm. Male and female SD - rats treated with zolmitriptan, dexamethasone and placebo （blank control） separately. And then animals received a single injection of midazolam （ 10 mg · kg^- 1 ） in the tail vein after the last dose of zolmitriptan, dexamethasone or placebo. The midazolam pharmacokinetics of zolmitriptan group, dexamethasone group and blank control were compared. Results. The standard curves for midazolam were linear in the range of 0. 3 - 375 μmol · L^- 1, r = 0. 9997, and the detection concentration of midazolam was 0. 0018 μmol · L^-1. The average recovery of midazolam was 97.9%. The area under the curve （ AUC0-t ） and the elimination half - life of midazolam was significantly reduced in male rats by the zolmitriptan and dexamethasone pretreatment. While in female rats there were no significantly changes. Conclusion： The method is stable,accurate and simple,and can be used for investigation the effect of zolmitriptan on CYP3A activity in rats. Zolmitriptan has the gender - different effect of induction on male rat hepatic CYP3A activity.

关 键 词：HPLC 咪达唑仑 佐米曲坦 诱导 CYP3A 

分 类 号：R917[医药卫生—药物分析学]