Internal standard tracked dilution to overcome challenges in dried blood spots and robotic sample preparation for liquid chromatography/tandem mass spectrometry assays
Internal standard tracked dilution to overcome challenges in dried blood spots and robotic sample preparation for liquid chromatography/tandem mass spectrometry assays
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DOI：
10.1002/rcm.4990
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发表时间：
2011-05-15
影响因子：
2
通讯作者：
Ji, Qin C.
中科院分区：
文献类型：
--
作者：
Liu, Guowen;Snapp, Heidi M.;Ji, Qin C.
干血斑（DBS）技术是一种新兴的生物分析样品采集方法。DBS样品的稀释是一个挑战，因为它是固体样品形式。目前，需要稀释的DBS样品首先作为常规样品提取，然后用含有内标物（IS）的提取空白样品稀释。由于稀释步骤是体积关键步骤，因此在处理有限体积的浸提样品时，必须格外小心以实现准确稀释。在这里，我们介绍了一种替代的样品稀释液液相色谱/串联质谱（LC/MS/MS）测定使用IS跟踪稀释步骤。在样品处理前，首先将稀释因子调整的IS工作溶液加入需要稀释的样品中;随后，在LC/MS/MS分析前，将处理后的样品近似稀释至试验线性响应范围内。我们将这种方法定义为“IS跟踪稀释”。这种方法的优点是稀释步骤由IS跟踪，不再是体积关键步骤。与样品稀释相关的另一个公认的挑战是使用液体处理器进行自动样品稀释。这种“IS跟踪稀释”还可以帮助解决液体样品的自动样品稀释的一些挑战。在使用奥美拉唑作为探针化合物的血浆测定和DBS测定中，这种新的稀释方法被证明是有效和方便的。版权所有（C）2011约翰威利父子有限公司
Dried blood spot (DBS) technology is an emerging alternative for sample collection in bioanalysis. Dilution for DBS samples is a challenge due to its solid sample format. Currently, DBS samples requiring dilution were first extracted as regular samples and then diluted with extracted blank samples containing internal standard (IS). Since the dilution step is a volume-critical step, extra care has to be taken to achieve accurate dilution when dealing with limited volume extracted samples. Here, we introduce an alternative sample dilution for liquid chromatography/tandem mass spectrometry (LC/MS/MS) assays using IS to track the dilution step. Dilution factor-adjusted IS working solution was first added to the sample requiring dilution before sample processing; subsequently, the processed sample was approximately diluted into the assay linear response range before LC/MS/MS analysis. We define this approach as "IS-tracked dilution". The advantage of this approach is that the diluting step is tracked by the IS and is no longer a volume-critical step. Another recognized challenge related to sample dilution is automatic sample dilution using a liquid handler. This "IS-tracked dilution" may also help address some of the challenges for automatic sample dilution of liquid samples. This new dilution approach was proven to be effective and convenient in both plasma assays and DBS assays using omeprazole as a probe compound. Copyright (C) 2011 John Wiley & Sons, Ltd.