Validation and clinical application of a novel LC-MS method for quantification of dolutegravir in breast milk
Validation and clinical application of a novel LC-MS method for quantification of dolutegravir in breast milk
复制标题
DOI：
10.4155/bio-2018-0085
复制
发表时间：
2018-12-01
期刊：
影响因子：
1.8
通讯作者：
Else, Laura J.
中科院分区：
文献类型：
--
作者：
Gini, Joshua;Penchala, Sujan Dilly;Else, Laura J.
目的：建立一种新的、灵敏的、可重复的测定母乳干斑中多替格拉韦（DTG）含量的方法，并验证其可用于临床研究。它的应用使母乳喂养的母亲及其婴儿的DTG药代动力学测量成为可能。结果/方法：样品采用液液萃取法，叔丁基甲醚萃取，DTG-d5为内标。DTG在反相C-18 Waters XBridge （3.5m: 2.1x50mm）柱上洗脱，梯度流动相由0.1%甲酸加去离子水或甲醇组成。在10-4000ng/ml的校准范围内验证该分析。结论：根据FDA和EMA生物分析方法指南，稳定性、测定间和测定内变异性是可接受的。该检测方法稳健、准确、精确，可可靠地应用于资源匮乏的临床试验样品的分析。
Aim: A novel, sensitive and reproducible method for quantification of dolutegravir (DTG) in dried breast milk spots (DBMS) was developed and validated for use in clinical studies. Its application enabled measurement of DTG pharmacokinetics in breastfeeding mothers and their infants. Results/methodology: Sample extraction was by liquid-liquid extraction using tert-butyl methy-ether, with DTG-d5 as an internal standard. DTG was eluted on a reverse phase C-18 Waters XBridge (3.5m: 2.1x50mm) column using a gradient mobile phase consisting of 0.1% formic acid in deionised water or methanol. The assay was validated over a calibration range of 10-4000ng/ml. Conclusion: Stability, inter and intra-assay variability were acceptable according to FDA and EMA bioanalytical method guidelines. The assay is robust, accurate, precise and can be reliably applied for analysis of clinical samples in trials from low resource settings.