LC-MS/MS法测定大鼠血浆内尼可地尔浓度及其药动学研究

目的： 建立LC-MS/MS法测定尼可地尔在大鼠血浆中的浓度，并初步研究其在大鼠体内的药动学行为。方法： 血浆样品以甲醇沉淀，采用Kromasil 100-5-C₁₈（2.1 mm×150 mm，5.0 μm）色谱柱进行色谱分离。以甲醇-含甲酸铵和甲酸的水溶液为流动相，流速0.3 mL·min^-1，进样量4 μL，LC-MS/MS分析，分别以m/z 212.1→136.0和m/z 271.2→172.2为尼可地尔和内标（甲苯磺丁脲）的质谱检测条件。大鼠灌服10 mg·kg^-1尼可地尔后，不同时间点取样测定其血浆中尼可地尔的浓度。由DAS 2.0计算药动学参数。结果： 尼可地尔质量浓度在5.0~2 000.0 ng·mL^-1内线性关系良好（r=0.997 9，权重1/X²）；定量下限为5.0 ng·mL^-1；尼可地尔和内标的提取回收率均高于95%，日内、日间的RSD均小于15%；尼可地尔血浆样品在室温放置4 h，-70℃冰箱放置15 d以及预处理后室温放置24 h的变化率均小于15%。大鼠口服灌胃尼可地尔（10 mg·kg^-1）后，尼可地尔在大鼠体内吸收较快，达峰时间约为0.3 h，达峰浓度约为15.0 μg·mL^-1。结论： 建立的LC-MS/MS方法适合于尼可地尔的药动学研究。

Objective: To establish a rapid and accurate method of LC-MS/MS determinates nicorandil concentration in rat plasma and to study its pharmacokinetics in rats. Methods: The plasma samples were precipitated with methanol and used the Kromasil 100-5-C₁₈ (2.1 mm×150 mm,5.0 μm) chromatographic column to separate nicorandil. The column temperature was 30℃. Methanol-the ammonium formate and formic acid aqueous solution was used as the mobile phase. The flow rate was 0.3 mL·min -1and sample volume was 4 μL. The detection conditions of nicorandil and internal standard (tolbutamide) by mass spectrometry were m/z 212.1-136.0 and m/z 271.2-136.0. Rats were given 10 mg·kg^-1 nicorandil by oral gavage and the plasma concentration of nicorandil was detected at different time points. Pharmacokinetic parameters were calculated by DAS 2.0 software. Results: The concentration of nicorandil had a good linear relationship between 5.0 and 2 000.0 ng·ml-1 (r=0.997 9,weights 1/X²). The quantitative lower limit was 5.0 ng·mL^-1. The extraction recovery of nicorandil and internal standard were higher than 95% and the RSDs of intra day and inter day were less than 15%. Nicorandil plasma samples were placed at room temperature for 4 h and refrigerated at -70℃for 15 days. Finally,the pretreatments and the rates of change are not less than 15% at room temperature for 24 h. After oral intragastric administration of nicodil (10 mg·kg^-1)to rats,the pharmacokinetic results showed that the peak time of nicodil in rats was about 0.3 h and the peak concentration was about 15.0 μg·mL^-1.Conclusion: The established LC-MS/MS method is suitable for studying the pharmacokinetic of nicorandil.

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