Automated high throughput analysis of antiretroviral drugs in dried blood spots
Automated high throughput analysis of antiretroviral drugs in dried blood spots
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DOI：
10.1002/jms.3952
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发表时间：
2017-08-01
影响因子：
2.3
通讯作者：
Haschke, M.
中科院分区：
文献类型：
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作者：
Duthaler, U.;Berger, B.;Haschke, M.
对于远程环境中的治疗药物监测，干血点（DBS）是特别有利的，因为血液样品收集和处理并不复杂。本研究的目的是开发和验证用于分析DBS样品中奈韦拉平、依法韦仑和洛匹那韦的自动提取方法。采用DBS-MS 500自动进样器和液相色谱串联质谱系统，以甲醇：水（70：30 v/v）进行自动提取。自动进样器使用每个DBS的数字图像定位提取头，将10 μ l内标物喷雾到每个DBS上，并使用25 μ l提取溶剂通过单向流动从每个斑点的中心提取4 mm圆盘（O）。在五氟苯基色谱柱上对分析物进行基线分离，并采用电喷雾电离和多反应监测进行分析，奈韦拉平和洛匹那韦采用正极性模式，依法韦仑采用负极性模式。该方法在10和10 000 ng/ml的所有分析物之间呈线性。自动样品提取导致不同供体和浓度水平之间的一致回收率（奈韦拉平：70 +/-6%，依法韦仑：63 +/- 11%和洛匹那韦：60 +/- 10%）和基质效应。日内和日间准确度和精密度偏差为
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