通过液相色谱-串联质谱（LC-MS/MS）分析对来自眼科泪液试纸和聚酯拭子的宫颈阴道分泌物中的达匹韦林和马拉维若进行双重定量。
Dual quantification of dapivirine and maraviroc in cervicovaginal secretions from ophthalmic tear strips and polyester-based swabs via liquid chromatographic-tandem mass spectrometric (LC-MS/MS) analysis.
作者信息
Parsons Teresa L, Emory Joshua F, Seserko Lauren A, Aung Wutyi S, Marzinke Mark A
机构信息
Department of Medicine, Johns Hopkins University, 600 North Wolfe Street, Osler 500, Baltimore, MD 21287, USA.
Department of Medicine, Johns Hopkins University, 600 North Wolfe Street, Osler 500, Baltimore, MD 21287, USA; Department of Pathology, Johns Hopkins University, 1800 Orleans Street, Sheikh Zayed Tower, B1020-G, Baltimore, MD 21287, USA.
出版信息
J Pharm Biomed Anal. 2014 Sep;98:407-16. doi: 10.1016/j.jpba.2014.06.018. Epub 2014 Jun 21.
BACKGROUND
Topical microbicidal agents are being actively pursued as a modality to prevent HIV viral transmission during sexual intercourse. Quantification of antiretroviral agents in specimen sources where antiviral activity is elicited is critical, and drug measurements in cervicovaginal fluid can provide key information on local drug concentrations. Two antiretroviral drugs, dapivirine and maraviroc, have gained interest as vaginal microbicidal agents, and rugged methods are required for their quantification in cervicovaginal secretions.
METHODS
Cervicovaginal fluid spiked with dapivirine and maraviroc were applied to ophthalmic tear strips or polyester-based swabs to mimic collection procedures used in clinical studies. Following sample extraction and the addition of isotopically labeled internal standards, samples were subjected to liquid chromatographic-tandem mass spectrometric (LC-MS/MS) analysis using a Waters BEH C8, 50mm×2.1mm, 1.7μm particle size column, on an API 4000 mass analyzer operated in selective reaction monitoring mode. The method was validated according to FDA Bioanalytical Method Validation guidelines.
RESULTS
Due to the disparate saturation capacity of the tested collection devices, the analytical measuring ranges for dapivirine and maravirocin cervicovaginal fluid on the ophthalmic tear strip were 0.05-25ng/tear strip, and 0.025-25ng/tear strip, respectively. As for the polyester-based swab, the analytical measuring ranges were 0.25-125ng/swab for dapivirine and 0.125-125ng/swab for maraviroc. Dilutional studies were performed for both analytes to extended ranges of 25,000ng/tear strip and 11,250ng/swab. Standard curves were generated via weighted (1/x(2)) linear or quadratic regression of calibrators. Precision, accuracy, stability and matrix effects studies were all performed and deemed acceptable according to the recommendations of the FDA Bioanalytical Method Validation guidelines.
CONCLUSIONS
A rugged LC-MS/MS method for the dual quantification of dapivirine and maraviroc in cervicovaginal fluid using two unique collection devices has been developed and validated. The described method meets the criteria to support large research trials.
背景
局部杀菌剂正被积极探索作为一种在性交过程中预防艾滋病毒传播的方式。在产生抗病毒活性的标本来源中对抗逆转录病毒药物进行定量至关重要，而宫颈阴道液中的药物测量可以提供有关局部药物浓度的关键信息。两种抗逆转录病毒药物，达匹韦林和马拉维罗，作为阴道杀菌剂受到关注，需要可靠的方法来对其在宫颈阴道分泌物中的含量进行定量。
方法
将添加了达匹韦林和马拉维罗的宫颈阴道液应用于眼科泪液试纸或聚酯拭子，以模拟临床研究中使用的采集程序。样品提取并添加同位素标记的内标后，使用 Waters BEH C8 柱（50mm×2.1mm，粒径 1.7μm），在以选择性反应监测模式运行的 API 4000 质谱仪上进行液相色谱 - 串联质谱（LC-MS/MS）分析。该方法根据美国食品药品监督管理局（FDA）生物分析方法验证指南进行验证。
结果
由于所测试的采集装置的饱和容量不同，达匹韦林和马拉维罗在眼科泪液试纸上宫颈阴道液中的分析测量范围分别为 0.05 - 25ng/泪液试纸和 0.025 - 25ng/泪液试纸。对于聚酯拭子，达匹韦林的分析测量范围为 0.25 - 125ng/拭子，马拉维罗为 0.125 - 125ng/拭子。对两种分析物都进行了稀释研究，将范围扩展至 25,000ng/泪液试纸和 11,250ng/拭子。通过校准物的加权（1/x²）线性或二次回归生成标准曲线。根据 FDA 生物分析方法验证指南的建议进行了精密度、准确度、稳定性和基质效应研究，结果均被认为可接受。
结论
已开发并验证了一种使用两种独特采集装置对宫颈阴道液中的达匹韦林和马拉维罗进行双重定量的可靠 LC-MS/MS 方法。所描述的方法符合支持大型研究试验的标准。
Zotero 插件