LC-MS/MS 法同时测定人血浆中比克替拉韦、度鲁特韦和拉替拉韦的验证。

Validation of an LC-MS/MS assay for the simultaneous determination of bictegravir, doravirine, and raltegravir in human plasma.

机构信息

Division of Pharmacotherapy and Experimental Therapeutics, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.

出版信息

J Pharm Biomed Anal. 2022 Oct 25;220:115010. doi: 10.1016/j.jpba.2022.115010. Epub 2022 Aug 20.

DOI:10.1016/j.jpba.2022.115010

PMID:36030752

Abstract

Bictegravir (BIC), an integrase inhibitor, and doravirine (DOR), a non-nucleoside reverse transcriptase inhibitor, were recently approved by the US FDA for HIV treatment and are recommended first line treatment options. Because certain clinical scenarios warrant using them in combination, we developed a fully validated LC-MS/MS method for simultaneous measurement of BIC and DOR, along with a legacy integrase inhibitor, raltegravir (RAL), in human plasma over a clinically relevant 1000-fold range for each analyte. These analytes were extracted from the plasma by protein precipitation with their stable, isotopically labeled internal standards (BIC-d, C-DOR, and RAL-d). Following extraction, samples were analyzed by reverse phase chromatography on a Waters Atlantis T3 C18 (50 ×2.1 mm, 3 µm particle size) column with subsequent detection by electrospray ionization in positive ion mode on an AB Sciex API-5000 triple quadrupole mass spectrometer. The assay was linear (R >0.994) over the selected calibration ranges (20.0-20,000 ng/mL (BIC), 3.00-3000 ng/mL (DOR), and 10.0-10,000 (RAL)). The assay was accurate (inter-assay %Bias ≤ ± 8.5) and precise (inter-assay %CV ≤11.4). This method was validated according to FDA guidance for industry and can be used to assess the pharmacokinetics of two newly approved antiretrovirals, or to support therapeutic drug monitoring for modern antiretroviral therapy.

摘要

比克替拉韦（BIC），一种整合酶抑制剂，和多拉韦林（DOR），一种非核苷类逆转录酶抑制剂，最近被美国 FDA 批准用于 HIV 治疗，是推荐的一线治疗选择。由于某些临床情况需要联合使用这些药物，我们开发了一种完全经过验证的 LC-MS/MS 方法，用于同时测量人血浆中的 BIC 和 DOR，以及一种传统的整合酶抑制剂，拉替拉韦（RAL），在每个分析物的临床相关 1000 倍范围内。这些分析物通过与稳定的、同位素标记的内标物（BIC-d、C-DOR 和 RAL-d）进行蛋白沉淀从血浆中提取出来。提取后，样品在 Waters Atlantis T3 C18（50×2.1mm，3μm 粒径）反相色谱柱上进行分析，随后在 AB Sciex API-5000 三重四极杆质谱仪上以正离子模式进行电喷雾电离检测。该测定法在选定的校准范围内（BIC：20.0-20,000ng/mL；DOR：3.00-3000ng/mL；RAL：10.0-10,000ng/mL）呈线性（R＞0.994）。该测定法准确（批内偏差≤±8.5%）且精密（批内变异系数≤11.4%）。该方法根据 FDA 行业指南进行了验证，可用于评估两种新批准的抗逆转录病毒药物的药代动力学，或支持现代抗逆转录病毒治疗的治疗药物监测。