Sensitive and specific LC-MS/MS method for the simultaneous measurements of viramidine and ribavirin in human plasma
Sensitive and specific LC-MS/MS method for the simultaneous measurements of viramidine and ribavirin in human plasma
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DOI：
10.1016/j.jchromb.2005.12.034
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发表时间：
2006-02-17
影响因子：
3
通讯作者：
Lin, CC
中科院分区：
文献类型：
--
作者：
Liu, YF;Xu, C;Lin, CC
Viramidine是利巴韦林的前药。为了便于进行人体中韦拉脒的药代动力学研究，开发并验证了一种灵敏且特异性高的LC-MS/MS方法，用于同时分析人血浆中的韦拉脒和利巴韦林。该方法包括加入[C-13]韦嘧啶和[C-13]利巴韦林作为内标物，用乙腈沉淀蛋白质，HPLC分离，并在多反应监测模式（MRM）下使用正电喷雾电离通过MS/MS系统进行定量。分别在245 -> 113、250 -> 113、244 -> 112和249 -> 112处监测利巴韦林、[C-13]利巴韦林、维拉米啶和[C-13]维拉米啶的前体->产物离子跃迁。维拉米定和病毒唑的校准曲线在1-1000 ng/mL的浓度范围内呈线性。对于韦拉脒和利巴韦林，定量下限（LLOQ）均为1 ng/mL。对于维拉米定，1、5、250和1000 ng/mL QC样品的日内和日间分析表明精密度（%CV在1.0 - 7.0%之间）和准确度（%偏倚在-4.3至5.2%之间）良好。对于利巴韦林，1、5、250和1000 ng/mL QC样品的日内和日间分析表明精密度（%CV在0.8和8.3%之间）和准确度（%偏倚在-5.8和9.4%之间）相似。在室温下储存至少3 h后，人血浆中的韦拉脒和利巴韦林均保持稳定。4 ℃至少6小时，并至少进行三次冻融循环。这种准确且高度特异性的测定方法为评价人体接受维拉米定给药后维拉米定和利巴韦林的药代动力学提供了一种有用的方法。(c)2006 Elsevier B. V.保留所有权利。
Viramidine is a prodrug of ribavirin. To facilitate pharmacokinetics studies of viramidine in man, a sensitive and specific LC-MS/MS method for the simultaneous analyses of viramidine and ribavirin in human plasma was developed and validated. The method involved the addition of [C-13]viramidine and [C-13]ribavirin as internal standards, protein precipitation with acetonitrile, HPLC separation, and quantification by MS/MS system using positive electrospray ionization in the multiple reaction monitoring mode (MRM). The precursor -> product ion transitions were monitored at 245 -> 113, 250 -> 113, 244 -> 112, and 249 -> 112 for ribavirin, [C-13]ribavifin, viramidine, and [C-13]viramidine, respectively. The calibration curves for viramidine and ribavirin were linear over a concentration range of 1-1000 ng/mL. For both viramidine and ribavirin, the lower limit of quantification (LLOQ) was 1 ng/mL. For viramidine, intra- and inter-day analyses of QC samples at 1, 5, 250, and 1000 ng/mL, indicated good precision (%CV between 1.0 and 7.0%) and accuracy (%bias between -4.3 and 5.2%). For ribavirin, intra- and inter-day analyses of QC samples at 1, 5, 250, and 1000ng/mL indicated similar precision (%CV between 0.8 and 8.3%) and accuracy (%bias between -5.8 and 9.4%). Both viramidine and ribavirin were stable in human plasma stored at room temperature for at least 3 h. 4 degrees C for at least 6 h, and for at least three freeze-thaw cycles. This accurate and highly specific assay provides a useful method for evaluating the pharmacokinetics of viramidine and ribavirin in man following administration of viramidine. (c) 2006 Elsevier B.V. All rights reserved.