Validation of a sensitive LC/MS/MS method for the determination of telaprevir and its R-isomer in human plasma.
Validation of a sensitive LC/MS/MS method for the determination of telaprevir and its R-isomer in human plasma.
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验证用于测定人血浆中特拉匹韦及其 R 异构体的灵敏 LC/MS/MS 方法。
DOI：
10.1002/bmc.3211
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发表时间：
2014
期刊：
影响因子：
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通讯作者：
Kiser,JenniferJ
中科院分区：
文献类型：
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作者：
Chen,Xinhui;Bushman,LaneR;McAllister,KevinJ;Anderson,PeterL;Kiser,JenniferJ
本研究的目的是验证反相高效液相色谱（HPLC）串联质谱（MS/MS）测定酸化和非酸化人血浆中替拉韦及其非对映体（VRT‐127394）的方法。telaprevir和telaprevir‐r的色谱基线分离采用Waters XBridgeTMBEH Shield C18色谱柱，2.1 × 75 mm，粒径2.5µm，流动相为水-乙腈-异丙醇：50:45:5,1%氨，0.2 mL/min，等压条件。该方法利用含有11个氘原子的稳定同位素内标对telaprevir分子（telaprevir‐d11）的结构进行测定。通过将telaprevir - d11在碱性溶液中孵育，制备了telaprevir - R - d11内标，促进了异构体的相互转化。采用正离子电喷雾（ESI+）质谱联用法对泰拉匹韦、泰拉匹韦‐R、泰拉匹韦‐IS和泰拉匹韦‐R‐IS进行检测和定量。提取酸化后的人血浆0.100 mL时，测定的定量限为5.0 ng/mL。在5.0-5000 ng/mL的校准范围内验证了准确度和精密度。使用患者样本证明，与先前的建议相反，特拉匹韦的定量不需要酸化血浆。版权所有©2014 John Wiley & Sons， Ltd。
The purpose of this study was to validate a reversed‐phase high‐performance liquid chromatographic (HPLC), tandem mass spectrometry (MS/MS) assay for the determination of telaprevir and itsR‐diastereomer (VRT‐127394) in acidified and nonacidified human plasma. The chromatographic baseline separation of telaprevir and telaprevir‐Rwas performed on a Waters XBridgeTMBEH Shield C18, 2.1 × 75 mm column with a 2.5 µm particle size, under isocratic conditions consisting of a mobile phase of 50:45:5 water–acetonitrile–isopropanol with 1% ammonia at 0.2 mL/min. This method utilized a stable isotope internal standard with 11 deuterium atoms on the structure of the telaprevir molecule (telaprevir‐d11). An internal standard for the telaprevir‐R(telaprevir‐R‐d11) was also prepared by incubating telaprevir‐d11 in basic solution, which facilitated isomer inter‐conversion. The detection and quantitation of telaprevir, telaprevir‐R, telaprevir‐IS and telaprevir‐R‐IS was achieved by positive ion electrospray (ESI+) MS/MS detection. The assay quantifiable limit was 5.0 ng/mL when 0.100 mL of acidified human plasma was extracted. Accuracy and precision were validated over the calibration range of 5.0–5000 ng/mL. It was demonstrated using patient samples that, contrary to previous recommendations, quantitation of telaprevir does not require acidified plasma. Copyright © 2014 John Wiley & Sons, Ltd.