Department of Chemistry, Purdue University, 560 Oval Drive, West Lafayette, IN, 47907, USA.
Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Denver, 12850 East Montview Blvd., Aurora, CO, 80045, USA.
Anal Bioanal Chem. 2020 Feb;412(6):1243-1249. doi: 10.1007/s00216-019-02304-0. Epub 2020 Jan 2.
Inadequate adherence to chronic medications is a far-reaching problem with financial and human health consequences. By a wide margin, non-adherence is the leading cause of therapeutic failures of HIV pre-exposure chemoprophylaxis (PrEP) and antiretroviral therapy (ART). It has been proven that HIV infection can be prevented by daily dosing of emtricitabine and tenofovir disoproxil fumarate. Measurement of intracellular tenofovir diphosphate in red blood cells has been established as an effective way to assess cumulative adherence, however, the LC-MS-based analytical method developed for the purpose is both complicated and expensive. Here, we report a simple method for adherence monitoring based on direct MS quantification of intracellular tenofovir diphosphate in human whole blood. The method requires only microliters of whole blood, employs special membranes to perform plasma separation and concomitant desalting during blood collection, and uses nanoelectrospray on a triple quadrupole instrument. Quantitative performance in this proof-of-concept study includes RSDs of < 15% and successful analysis of a small number of patient samples with medium to high adherence levels. The results correlate with those of a validated LC-MS/MS method, and an R value of 0.9962 is achieved. This methodology has promise for extension to point-of-care testing using miniature mass spectrometers. Graphical abstract.
慢性药物治疗依从性不足是一个具有深远影响的问题，会带来经济和人类健康方面的后果。很大程度上，不依从是导致 HIV 暴露前预防 (PrEP) 和抗逆转录病毒治疗 (ART) 治疗失败的主要原因。每天服用恩曲他滨和替诺福韦二吡呋酯已被证明可以预防 HIV 感染。已经证实，通过测量红细胞内的替诺福韦二磷酸来评估累积的药物依从性是一种有效的方法，然而，为此目的开发的基于 LC-MS 的分析方法既复杂又昂贵。在这里，我们报告了一种基于人全血中替诺福韦二磷酸的直接 MS 定量的简单药物依从性监测方法。该方法仅需要微升全血，在采集血液时使用特殊的膜进行血浆分离和同时脱盐，并在三重四极杆仪器上进行纳升电喷雾。在这项概念验证研究中，定量性能包括 <15%的 RSD 和对具有中到高依从性水平的少数患者样本的成功分析。结果与经过验证的 LC-MS/MS 方法相关，达到了 0.9962 的 R 值。该方法有望扩展到使用微型质谱仪的即时检测。
