Nucleoside reverse transcriptase inhibitors (NRTIs) have been the first class of antiretroviral drugs used against HIV infection. Despite NRTI-free regimens have been eagerly sought over the years in an effort for treatment simplification, NRTIs remain in most antiretroviral combination treatment. There has been generally a limited interest for their therapeutic drug monitoring, arguably because NRTIs levels measured in plasma poorly predict the concentration of pharmacologically active metabolites in cells. Plasma concentrations do impact cellular levels, while large differences between NRTIs have been found with regard to their ability to distribute into the cerebrospinal fluid (CSF) compartment. The renewed interest for the measurements of NRTIs concentrations in plasma and CSF was raised by ongoing efforts to understand some instances of toxicity or for determining their actual implication in the development of HIV-associated neurological disorders. In this context, a 5-min multiplex ultra-high-pressure chromatography tandem mass spectrometry (UHPLC-MS/MS) analysis in human plasma and CSF was developed for NRTIs used in clinical practice: abacavir, emtricitabine, lamivudine, tenofovir and zidovudine along with zidovudine glucuronide (Gln-ZDV). The 200-fold dilution of blank human plasma was shown to be a reliable surrogate matrix for quantification of NRTIs and Gln-ZDV in CSF. Both methodologies were fully validated over the clinically relevant concentrations, and satisfactorily fulfilled all parameters for bioanalytical methods validation. This sensitive, rapid, and robust UHPLC-MS/MS assay offers a methodology for increasing our understanding of the ability of NRTIs to cross the blood-brain barrier and their potential implication in neuropsychological disorders observed in HIV-infected patients.

核苷逆转录酶抑制剂（NRTI）是第一类用于对抗 HIV 感染的抗逆转录病毒药物。尽管多年来为了简化治疗而迫切寻求不含 NRTI 的治疗方案，但 NRTI 仍然存在于大多数抗逆转录病毒联合治疗中。人们普遍对其治疗药物监测兴趣有限，可能是因为血浆中测量的 NRTI 水平很难预测细胞中药理活性代谢物的浓度。血浆浓度确实会影响细胞水平，而 NRTI 之间分布到脑脊液 (CSF) 区室的能力存在很大差异。为了了解某些毒性实例或确定其在 HIV 相关神经系统疾病发展中的实际影响，人们不断努力了解血浆和脑脊液中 NRTI 浓度的测量，这引起了人们对测量血浆和脑脊液中 NRTI 浓度的新兴趣。在此背景下，针对临床实践中使用的 NRTI 开发了人血浆和脑脊液中的 5 分钟多重超高压色谱串联质谱 (UHPLC-MS/MS) 分析：阿巴卡韦、恩曲他滨、拉米夫定、替诺福韦和齐多夫定与齐多夫定葡萄糖苷酸 (Gln-ZDV) 一起使用。 200 倍稀释的空白人血浆被证明是定量 CSF 中 NRTI 和 Gln-ZDV 的可靠替代基质。两种方法都在临床相关浓度上得到了充分验证，并令人满意地满足了生物分析方法验证的所有参数。 这种灵敏、快速且稳健的 UHPLC-MS/MS 检测提供了一种方法，可增强我们对 NRTI 穿越血脑屏障的能力及其对 HIV 感染患者中观察到的神经心理疾病的潜在影响的了解。