Development and validation of an LC-MS/MS assay for the quantification of dolutegravir extracted from human hair.
Development and validation of an LC-MS/MS assay for the quantification of dolutegravir extracted from human hair.
复制标题
开发和验证 LC-MS/MS 测定法，用于定量从人发中提取的多替拉韦。
DOI：
10.1007/s00216-018-1394-y
复制
发表时间：
2018
影响因子：
4.3
通讯作者：
Kashuba,AngelaDM
中科院分区：
文献类型：
--
作者：
Sykes,Craig;Blake,Kimberly;White,Nicole;Schauer,AmandaP;Guzman,BryanB;Cottrell,MackenzieL;Tamraz,Bani;Kashuba,AngelaDM
毛发中药物浓度的测量提供了评估药物依从性的非侵入性方法。在这里，我们报告的发展和验证的方法，用于定量的抗逆转录病毒dolutegravir（DTG）从人的头发中提取。通过超声处理从毛发样品中提取DTG，并在40 °C下在含有2%甲酸的50：50甲醇：乙腈中孵育过夜。提取后，在沃茨Atlantis T3（50 × 2.1 mm，3 μm粒径）色谱柱上通过反相色谱法分析样品，随后在AB Sciex API-5000三重四极杆质谱仪上以正离子模式通过电喷雾电离进行检测。稳定的同位素标记的13 C，d5-DTG用作测定中的内标。校准范围为5- 10，000 pg DTG/mL浸提溶剂，能够浸提1 - 10 mg毛发/mL浸提溶剂。该试验具有线性、准确度（试验间%偏倚在± 6.5%范围内）和精密度（试验间%CV ≤ 10.3%）。该检测试剂盒已成功用于分析DTG方案受试者的临床样本。临床样品分析表明可能存在降解产物，随后证实其在暴露于阳光下时发生。DTG的降解可能会使临床结果的绝对解释复杂化，但该降解产物的存在很容易用该测定法进行评价，以帮助数据解释。
Measurement of drug concentrations in hair provides a non-invasive approach to assess drug adherence. Here, we report on the development and validation of a method for the quantification of the antiretroviral dolutegravir (DTG) extracted from human hair. DTG is extracted from hair samples by sonication and incubation in 50:50 methanol:acetonitrile with 2% formic acid overnight at 40 °C. Following extraction, samples are analyzed by reverse-phase chromatography on a Waters Atlantis T3 (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 stable, isotopically labeled13C,d5-DTG is used as an internal standard in the assay. The calibration range is 5–10,000 pg DTG/mL of extraction solvent with the ability to extract between 1 and 10 mg of hair/mL of extraction solvent. The assay was linear, accurate (inter-assay %bias within ± 6.5%), and precise (inter-assay %CV ≤ 10.3%). The assay was successfully used to analyze clinical samples from subjects on DTG regimens. Analysis of clinical samples suggested the potential presence of a degradation product, which was subsequently confirmed to occur with exposure to sunlight. The degradation of DTG could complicate absolute interpretation of clinical results, but the presence of this degradation product is easily evaluated with this assay to aid in data interpretation.