Laboratory of Neuro-Oncology and Clinical and Cancer Proteomics, Department of Neurology, University Medical Center Rotterdam, Dr. Molewaterplein 50, 3015 GE Rotterdam, The Netherlands.
J Mass Spectrom. 2011 Mar;46(3):282-9. doi: 10.1002/jms.1897.
A new and reliable mass spectrometric method using an isotope dilution method in combination with matrix-assisted laser desorption/ionization-triple quadrupole tandem mass spectrometry (ID-MALDI-QqQ-MS/MS) has been developed and validated for the determination of concentrations of the antiretroviral drug tenofovir (TNV) in plasma from HIV-infected adults. The advantage of this new method is that (1) the method is ultrafast and (2) can be applied for high-throughput measurement of TNV in plasma. The method is based on a simple plasma deproteinization step in combination with the use of [adenine-(13) C(5) ]-TNV as the internal standard. TNV and [adenine-(13) C(5) ]-TNV were monitored by multiple reaction monitoring using the transition m/z 288.0 → 176.2 and m/z 293.2 → 181.2 for TNV and [adenine-(13) C(5) ]-TNV, respectively. The method was validated according to the most recent FDA guidelines for the development and validation of (new) bio-analytical assays. Validated method parameters were: linearity, accuracy, precision and stability of the method. The lowest limit of quantification was 0.10 µmol/l, whereas the limit of detection determined at a signal-to-noise ratio (S/N = 3:1) in pooled drug free human control plasma was 0.04 µmol/l. The validated method was successfully applied and tested for its clinical feasibility by the analysis of plasma samples from selected HIV-infected adults receiving the prodrug tenofovir disoproxil fumarate. Observed plasma TNV concentrations ranged between 0.11 and 0.76 µmol/l and measured plasma TNV concentrations were within the therapeutically relevant concentration range.
已开发并验证了一种新的、可靠的质谱方法，采用同位素稀释法结合基质辅助激光解吸/电离-三重四极杆串联质谱法（ID-MALDI-QqQ-MS/MS），用于测定艾滋病毒感染成人血浆中抗逆转录病毒药物替诺福韦（TNV）的浓度。这种新方法的优点是：(1) 方法超快速；(2) 可用于高通量测量血浆中的 TNV。该方法基于简单的血浆去蛋白化步骤，并结合使用 [腺嘌呤-(13)C(5)]-TNV 作为内标。TNV 和 [腺嘌呤-(13)C(5)]-TNV 通过多重反应监测，使用 m/z 288.0→176.2 和 m/z 293.2→181.2 的跃迁分别进行监测。该方法按照 FDA 最近发布的关于（新）生物分析测定法的开发和验证指南进行了验证。验证的方法参数包括：方法的线性、准确度、精密度和稳定性。定量下限为 0.10 µmol/l，而在混合无药物的人对照血浆中，信噪比（S/N=3:1）确定的检测限为 0.04 µmol/l。该验证方法成功地应用于分析接受前药替诺福韦酯富马酸二异丙酯的选定艾滋病毒感染成人的血浆样本，并测试了其临床可行性。观察到的血浆 TNV 浓度范围为 0.11 至 0.76 µmol/l，测量的血浆 TNV 浓度处于治疗相关浓度范围内。
