Rackow Ashley R, Pandey Aashish, Price Amelia L, Marzinke Mark A
Division of Clinical Chemistry, Department of Pathology, Johns Hopkins University, United States.
Division of Clinical Pharmacology, Department of Medicine, Johns Hopkins University, United States.
J Mass Spectrom Adv Clin Lab. 2024 Sep 19;34:1-7. doi: 10.1016/j.jmsacl.2024.09.001. eCollection 2024 Nov.
Dolutegravir (DTG) is part of a first-line antiretroviral therapy (ART) for HIV management in drug-naïve individuals and is recommended for the treatment of HIV during pregnancy. Robust analytical tools to quantify DTG are necessary to support clinical trials that characterize its multi-compartment drug distribution.
Potassium EDTA (KEDTA) plasma or whole breast milk was spiked with DTG and an isotopically labeled internal standard. Samples were prepared via protein precipitation prior to LC-MS/MS analysis. The assays were validated in accordance with regulatory recommendations.
Analytical measuring ranges for DTG quantitation in plasma and breast milk were 100-10,000 ng/mL and 0.500 to 1000 ng/mL, respectively. Inter-assay precision and accuracy were 2.73 % to 3.41 % and -10.6 % to -5.37 % for plasma, and 4.24 % to 12.4 % and -5.63 % to 7.49 % for breast milk, respectively. DTG was stable for three freeze-thaw cycles and for at least 72 h at room temperature in matrix (plasma or breast milk). Additionally, whole blood was stable for 24 h at room temperature and 2 h under conditions of extended heat and humidity. Matrix effects for DTG in plasma and breast milk ranged from 101 % to 108 % and 78.2 % to 99.3 %, respectively. Quantitation in remnant plasma samples yielded measurable concentrations within the primary linearity of the assay.
Methods to quantify DTG in human plasma and breast milk have been developed and validated. These assays were designed to satisfy all criteria for implementation in clinical and clinical trial settings.
多替拉韦（DTG）是初治个体HIV管理一线抗逆转录病毒疗法（ART）的一部分，并且被推荐用于妊娠期HIV的治疗。强大的用于定量DTG的分析工具对于支持表征其多房室药物分布的临床试验是必要的。
向乙二胺四乙酸钾（KEDTA）血浆或全母乳中加入DTG和一种同位素标记的内标。在进行液相色谱-串联质谱（LC-MS/MS）分析之前，通过蛋白质沉淀法制备样品。这些检测方法按照监管建议进行了验证。
血浆和母乳中DTG定量的分析测量范围分别为100 - 10,000 ng/mL和0.500至1000 ng/mL。血浆的批间精密度和准确度分别为2.73%至3.41%和 - 10.6%至 - 5.37%，母乳的批间精密度和准确度分别为4.24%至12.4%和 - 5.63%至7.49%。DTG在基质（血浆或母乳）中经三个冻融循环以及在室温下至少72小时均稳定。此外，全血在室温下稳定24小时，在高温高湿条件下稳定2小时。血浆和母乳中DTG的基质效应范围分别为1??1%至108%和78.2%至99.3%。残余血浆样品中的定量在检测的主要线性范围内产生了可测量的浓度。
已开发并验证了定量人血浆和母乳中DTG的方法。这些检测方法旨在满足临床和临床试验环境中实施的所有标准。
