反相高效液相色谱法测定大鼠血浆中6,7-二甲氧基香豆素含量及其在药动学中的应用

RP-HPLC method for determination of 6,7-dimethoxycoumarin in rat plasma and its application to pharmacokinetics

目的 :建立大鼠血浆中 6 ,7 二甲氧基香豆素反相高效液相色谱紫外检测方法 ,并研究其在大鼠体内的药动学过程。方法 :血浆样品经乙腈沉淀蛋白 ,上清液在 5 0℃下经氮气流吹干 ,残渣加入流动相 10 0 μL溶解后 ,离心 ,直接进样。流动相为甲醇 1%乙酸水溶液 四氢呋喃 (30∶6 3∶7)。检测波长为 340nm。大鼠经灌胃予 6 ,7 二甲氧基香豆素后 ,所得血药浓度数据用Topfit程序拟合并估算药动学参数。结果 :线性范围为 0 0 2 5～ 2 5 μg·mL-1。经非房室模型计算 ,6 ,7 二甲氧基香豆素主要药动学参数T1/ 2 为 0 7h ,AUC0→t为 16 5 0 6ng·h·mL-1,AUC0→∞ 为 172 8 9ng·h·mL-1,CLtot为 5 4 0L·h-1,用Topfit程序拟合 ,其体内过程符合二室开放模型。结论 :本法操作简便 ,可应用于大鼠血浆中 6 ,7 二甲氧基香豆素浓度的测定及其药动学研究。

AIM: To develop a reversed phase high performance liquid chromatography ultraviolet method to determine 6,7 dimethoxycoumarin in rat plasma and to study the pharmacokinetic process of 6,7 dimethoxycoumarin in rats. METHODS: The drug was extracted with acetonitrile from plasma and separated on a Kromasil ODS (250 mm×4 6 mm,5 μm)column with methanol 1% solution of acetic acid tetrahydrofuran (30∶63∶7) as mobile phase, UV detector was set at 340 nm. The date obtained were analyzed with Topfit program. RESULTS: The linear calibration curves were obtained in the concentration range of 0.025-2 50 μg·mL -1 . With non compartmental calculation,the main pharmacokinetic parameters were as follow: T 1/2 was 0.7h, AUC 0→t was 1 650.6 ng·h·mL -1 , AUC 0→∞ was 1 728.9 ng·h·mL -1 , CL tot was 54.0 L·h -1 . Plasma concentration time profile after oral administration of 6,7 dimethoxycoumarin was fitted in with two compartment model. CONCLUSION: The method is simple and accurate and can be used to determin the concentration of 6,7 dimethoxycoumarin in plasma samples and to study pharmacokinetics in rats.