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Automated high throughput analysis of antiretroviral drugs in dried blood spots

机译：干血斑抗逆转录病毒药物自动化高通量分析

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摘要

For therapeutic drug monitoring in remote settings, dried blood spots (DBS) are particularly advantageous, as blood sample collection and handling is uncomplicated. The aim of this study was to develop and validate an automated extraction method for the analysis of nevirapine, efavirenz and lopinavir in DBS samples. Automated extractionwas performed with methanol : water (70 : 30 v/v), using a DBS-MS 500 autosampler coupled to a liquid chromatography tandem mass spectrometry system. The autosampler used digital images of each DBS to position the extraction head, sprayed 10 mu l of internal standard onto each DBS and extracted a 4-mm disc (O) from the centre of each spot by unilateral flow using 25-mu l extraction solvent. The analytes were baseline separated on a pentafluorophenyl column and analysed by using electrospray ionization with multiple reaction monitoring in positive polarity mode for nevirapine and lopinavir and in negative mode for efavirenz. The method was linear between 10 and 10 000 ng/ml for all analytes. Automated sample extraction resulted in consistent recoveries (nevirapine: 70 +/- 6%, efavirenz: 63 +/- 11% and lopinavir: 60 +/- 10%) and matrix effects between different donors and concentration levels. Intra-day and inter-day accuracy and precision deviations were <= 15%. Manual and automated extractions of DBS samples collected within the framework of an adherence assessment study in rural Tanzania showed good agreements with deviations of less than 10%. Our study highlights that therapeutic drug monitoring samples obtained in the resource-constrained setting of rural Africa can be reliably determined by automated extraction of DBS. Overall, automatization improved method sensitivity and facilitates analysis of large sample numbers. Copyright (C) 2017 John Wiley & Sons, Ltd.
机译：对于远程环境下的治疗药物监测，干血点（DBS）尤其有利，因为血样采集和处理非常简单。本研究的目的是开发并验证一种自动提取方法，用于分析DBS样品中的奈韦拉平、依法韦伦和洛匹那韦。使用DBS-MS 500自动进样器和液相色谱-串联质谱系统，使用甲醇：水（70:30 v/v）进行自动萃取。自动取样器使用每个DBS的数字图像来定位提取头，在每个DBS上喷洒10μl内标物，并使用25μl提取溶剂通过单向流动从每个斑点的中心提取一个4 mm圆盘（O）。在五氟苯基柱上对分析物进行基线分离，并在奈韦拉平和洛匹那韦的正极性模式下和依法韦伦的负极性模式下使用电喷雾电离和多反应监测进行分析。对于所有分析物，该方法在10至10000 ng/ml之间呈线性。自动样品提取导致了不同供体和浓度水平之间的一致回收率（奈韦拉平：70+/-6%，依法韦伦：63+/-11%和洛匹那韦：60+/-10%）和基质效应。日内和日间的准确度和精密度偏差<=15%。在坦桑尼亚农村进行的一项依从性评估研究的框架内，对DBS样本的手动和自动提取显示出良好的一致性，偏差小于10%。我们的研究强调，在非洲农村资源有限的环境中获得的治疗药物监测样本可以通过DBS的自动提取可靠地确定。总的来说，自动化提高了方法的灵敏度，并有助于分析大量样本。版权所有（C）2017约翰威利父子有限公司。

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  • 来源
    《Journal of mass spectrometry: JMS》 |2017年第8期|共9页
  • 作者

    Duthaler U.; Berger B.; Erb S.; Battegay M.; Letang E.; Gaugler S.; Krahenbuhla S.; Haschke M.;

  • 作者单位

    Univ Basel Univ Hosp Basel Dept Biomed &

    Clin Res Div Clin Pharmacol &

    Toxicol Hebelstr 20 CH-4031 Basel Switzerland;

    Univ Basel Univ Hosp Basel Dept Biomed &

    Clin Res Div Clin Pharmacol &

    Toxicol Hebelstr 20 CH-4031 Basel Switzerland;

    Univ Basel Univ Hosp Basel Dept Med &

    Clin Res Div Infect Dis &

    Hosp Epidemiol Petersgraben 4 CH-4031 Basel Switzerland;

    Univ Basel Univ Hosp Basel Dept Med &

    Clin Res Div Infect Dis &

    Hosp Epidemiol Petersgraben 4 CH-4031 Basel Switzerland;

    Univ Basel Swiss Trop &

    Publ Hlth Inst Med Dept Clin Res Unit Socinstr 57 CH-4051 Basel Switzerland;

    CAMAG Sonnenmattstr 11 CH-4132 Muttenz Switzerland;

    Univ Basel Univ Hosp Basel Dept Biomed &

    Clin Res Div Clin Pharmacol &

    Toxicol Hebelstr 20 CH-4031 Basel Switzerland;

    Univ Basel Univ Hosp Basel Dept Biomed &

    Clin Res Div Clin Pharmacol &

    Toxicol Hebelstr 20 CH-4031 Basel Switzerland;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 分析化学;
  • 关键词

    nevirapine; efavirenz; lopinavir; liquid chromatography tandem mass spectrometry; dried blood spots; automated extraction;

    机译：None;

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