Does volumetric absorptive microsampling eliminate the hematocrit bias for caffeine and paraxanthine in dried blood samples? A comparative study
Does volumetric absorptive microsampling eliminate the hematocrit bias for caffeine and paraxanthine in dried blood samples? A comparative study
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DOI：
10.1016/j.aca.2015.04.056
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发表时间：
2015-06-30
影响因子：
6.2
通讯作者：
Stove, Christophe P.
中科院分区：
文献类型：
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作者：
De Kesel, Pieter M. M.;Lambert, Willy E.;Stove, Christophe P.
体积吸收微量取样（VAMS）是一种新的取样技术，它可以直接从一滴或一池血液中采集精确体积的血液（约10 μ L），方法是将一个吸收性聚合物尖端浸入其中，然后将所得血液微量样品干燥并作为一个整体进行分析。本研究的目的是评估VAMS克服红细胞压积偏倚的潜力，这是干血微量样品分析中的一个重要问题。用于分析VAMS样品中模型化合物咖啡因和副黄嘌呤的LC-MS/MS方法得到充分验证，并符合所有预定标准。结合先前验证的干血斑（DBS）和血液的程序，这使我们能够建立一个细致的比较研究，其中两种化合物在80多个相应的VAMS，DBS和液体全血样品中测定。这些来源于真实的人类患者样本，覆盖广泛的红细胞比容范围（0.21-0.50）。通过计算与参考全血浓度的差异，我们发现VAMS样本中的分析物浓度不受在评价的红细胞压积范围内变化的偏倚的影响，这与DBS结果相反。然而，VAMS浓度倾向于高估全血浓度，因为观察到一致的正偏倚。从真实血液和加标血液制备的VAMS样本的不同行为，以及在高血细胞比容值下VAMS尖端的咖啡因和副黄嘌呤回收率略有降低，使用非常相似的提取程序未观察到DBS的影响，被认为是观察到的VAMS-全血偏差的基础。基于这项研究，作为第一个通过分析真实人体样本来评估VAMS的有效性和稳健性的研究，可以得出结论，VAMS有效地帮助消除了红细胞压积的影响。(C)2015作者由爱思唯尔公司出版
Volumetric absorptive microsampling (VAMS) is a novel sampling technique that allows the straightforward collection of an accurate volume of blood (approximately 10 mu L) from a drop or pool of blood by dipping an absorbent polymeric tip into it. The resulting blood microsample is dried and analyzed as a whole. The aim of this study was to evaluate the potential of VAMS to overcome the hematocrit bias, an important issue in the analysis of dried blood microsamples. An LC-MS/MS method for analysis of the model compounds caffeine and paraxanthine in VAMS samples was fully validated and fulfilled all pre-established criteria. In conjunction with previously validated procedures for dried blood spots (DBS) and blood, this allowed us to set up a meticulous comparative study in which both compounds were determined in over 80 corresponding VAMS, DBS and liquid whole blood samples. These originated from authentic human patient samples, covering a wide hematocrit range (0.21-0.50). By calculating the differences with reference whole blood concentrations, we found that analyte concentrations in VAMS samples were not affected by a bias that changed over the evaluated hematocrit range, in contrast to DBS results. However, VAMS concentrations tend to overestimate whole blood concentrations, as a consistent positive bias was observed. A different behavior of VAMS samples prepared from incurred and spiked blood, combined with a somewhat reduced recovery of caffeine and paraxanthine from VAMS tips at high hematocrit values, an effect that was not observed for DBS using a very similar extraction procedure, was found to be at the basis of the observed VAMS-whole blood deviations. Based on this study, being the first in which the validity and robustness of VAMS is evaluated by analyzing incurred human samples, it can be concluded that VAMS effectively assists in eliminating the effect of hematocrit. (C) 2015 The Authors. Published by Elsevier B.V.