稳定同位素内标对人血浆样品中伊伐布雷定和N-去甲基伊伐布雷定测定的影响

发布时间：2018-04-12 08:03

本文选题：稳定同位素内标 + LC-MS/MS　；参考：《药学学报》2015年03期

【摘要】：建立液相色谱-串联质谱(LC-MS/MS)法同时测定人血浆中伊伐布雷定和N-去甲基伊伐布雷定,并考察稳定同位素内标d3-伊伐布雷定对待测物测定的影响。血浆样品经乙腈沉淀蛋白处理后,经Capcell PAK C18柱(100 mm×4.6 mm,5μm)分离,以甲醇-5 mmol·L-1醋酸铵溶液为流动相。采用电喷雾电离源(ESI源),多反应监测模式进行正离子检测。实验中考察了伊伐布雷定同位素峰[M+H+3]+对内标峰面积的影响以及内标的同位素纯度对伊伐布雷定测定的干扰,通过选择合适的内标浓度满足了方法选择性和线性范围的要求。方法验证结果表明,测定伊伐布雷定和N-去甲基伊伐布雷定线性范围分别为0.100~60.0 ng·m L-1和0.050 0~20.0ng·m L-1。各待测物的日内、日间精密度以及准确度均符合生物样品分析相关要求。本方法成功应用于硫酸氢伊伐布雷定缓释片的人体药动学研究。
[Abstract]:A liquid chromatography-tandem mass spectrometry (LC-MS / MS) method was developed for the simultaneous determination of Ifabradine and N-demethyl-ivalburetine in human plasma, and the influence of the stable isotope internal standard d3-Ifabradine on the determination of the determinants was investigated.Plasma samples were treated with acetonitrile precipitated protein and separated by Capcell PAK C18 column (100mm 脳 4.6mm ~ 5 渭 m). The mobile phase was methanol-5 mmol L-1 ammonium acetate solution.The electrospray ionization source (ESI) was used to detect positive ions in multi-reaction monitoring mode.The influence of the Ifabradine isotope peak [MH3] on the area of the internal standard peak and the interference of the internal standard isotope purity on the determination of Ifabradine were investigated. The method selectivity and linear range were satisfied by selecting the appropriate concentration of the internal standard.The results showed that the linear ranges for the determination of Ifabradine and N- demethyllfabradine were 0.100,60.0 ng mL ~ (-1) and 0.050 0~20.0ng / mL ~ (-1), respectively.The day precision and accuracy of each object met the requirements of biological sample analysis.The method has been successfully applied to the pharmacokinetic study of Ephabradine sulfate sustained-release tablets.
【作者单位】：中国科学院上海药物研究所;中国药科大学药学院;
【分类号】：R96

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